Journal of Clinical Medicine

16 pages, 5173 KiB

Open AccessArticle

Maturity-Onset Diabetes of the Young (MODY) in Portugal: Novel GCK, HNFA1 and HNFA4 Mutations

by Maria I. Alvelos, Catarina I. Gonçalves, Eduarda Coutinho, Joana T. Almeida, Margarida Bastos, Maria L. Sampaio, Miguel Melo, Sofia Martins, Isabel Dinis, Alice Mirante, Leonor Gomes, Joana Saraiva, Bernardo D. Pereira, Susana Gama-de-Sousa, Carolina Moreno, Daniela Guelho, Diana Martins, Carla Baptista, Luísa Barros, Mara Ventura, Maria M. Gomes and Manuel C. Lemos Show full author list Hide full author list

J. Clin. Med. 2020, 9(1), 288; https://doi.org/10.3390/jcm9010288 - 20 Jan 2020

Cited by 10 | Viewed by 6123

Abstract

Maturity-onset diabetes of the young (MODY) is a frequently misdiagnosed type of diabetes, which is characterized by early onset, autosomal dominant inheritance, and absence of insulin dependence. The most frequent subtypes are due to mutations of the GCK (MODY 2), HNF1A (MODY 3), [...] Read more.

Maturity-onset diabetes of the young (MODY) is a frequently misdiagnosed type of diabetes, which is characterized by early onset, autosomal dominant inheritance, and absence of insulin dependence. The most frequent subtypes are due to mutations of the GCK (MODY 2), HNF1A (MODY 3), and HNF4A (MODY 1) genes. We undertook the first multicenter genetic study of MODY in the Portuguese population. The GCK, HNF1A, and HNF4A genes were sequenced in 46 unrelated patients that had at least two of the three classical clinical criteria for MODY (age at diagnosis, family history, and clinical presentation). The functional consequences of the mutations were predicted by bioinformatics analysis. Mutations were identified in 23 (50%) families. Twelve families had mutations in the GCK gene, eight in the HNF1A gene, and three in the HNF4A gene. These included seven novel mutations (GCK c.494T>C, GCK c.563C>G, HNF1A c.1623G>A, HNF1A c.1729C>G, HNF4A c.68delG, HNF4A c.422G>C, HNF4A c.602A>C). Mutation-positive patients were younger at the time of diagnosis when compared to mutation-negative patients (14.3 vs. 23.0 years, p = 0.011). This study further expands the spectrum of known mutations associated with MODY, and may contribute to a better understanding of this type of diabetes and a more personalized clinical management of affected individuals. Full article

(This article belongs to the Section Endocrinology & Metabolism)

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(a–d) Families with identified maturity-onset diabetes of the young (MODY) mutations. Filled symbols represent patients with diabetes, open symbols represent unaffected individuals. Squares, circles, and diamond symbols denote males, females, and unspecified, respectively. Numbers within symbols indicate additional siblings with the same phenotype. Oblique lines through symbols represent deceased individuals. Arrows indicate the index cases. The age of diagnosis of diabetes (y, years), when known, is presented. The presence (+) or absence (–) of the mutation, when known, is presented. The chromatograms of the DNA sequence for normal individuals and for patients with mutations (asterisks) are presented below each pedigree. Full article ">Figure 1 Cont.
(a–d) Families with identified maturity-onset diabetes of the young (MODY) mutations. Filled symbols represent patients with diabetes, open symbols represent unaffected individuals. Squares, circles, and diamond symbols denote males, females, and unspecified, respectively. Numbers within symbols indicate additional siblings with the same phenotype. Oblique lines through symbols represent deceased individuals. Arrows indicate the index cases. The age of diagnosis of diabetes (y, years), when known, is presented. The presence (+) or absence (–) of the mutation, when known, is presented. The chromatograms of the DNA sequence for normal individuals and for patients with mutations (asterisks) are presented below each pedigree. Full article ">Figure 1 Cont.
(a–d) Families with identified maturity-onset diabetes of the young (MODY) mutations. Filled symbols represent patients with diabetes, open symbols represent unaffected individuals. Squares, circles, and diamond symbols denote males, females, and unspecified, respectively. Numbers within symbols indicate additional siblings with the same phenotype. Oblique lines through symbols represent deceased individuals. Arrows indicate the index cases. The age of diagnosis of diabetes (y, years), when known, is presented. The presence (+) or absence (–) of the mutation, when known, is presented. The chromatograms of the DNA sequence for normal individuals and for patients with mutations (asterisks) are presented below each pedigree. Full article ">Figure 1 Cont.
(a–d) Families with identified maturity-onset diabetes of the young (MODY) mutations. Filled symbols represent patients with diabetes, open symbols represent unaffected individuals. Squares, circles, and diamond symbols denote males, females, and unspecified, respectively. Numbers within symbols indicate additional siblings with the same phenotype. Oblique lines through symbols represent deceased individuals. Arrows indicate the index cases. The age of diagnosis of diabetes (y, years), when known, is presented. The presence (+) or absence (–) of the mutation, when known, is presented. The chromatograms of the DNA sequence for normal individuals and for patients with mutations (asterisks) are presented below each pedigree. Full article ">Figure 2
Schematic representation of the GCK, HNF1A, and HNF4A genes and positions of the identified mutations. Numbered boxes represent exons and lines represent introns (not to scale). UTR, untranslated regions. A 250 base-pair (bp) scale is shown. Full article ">

60 pages, 428 KiB

Open AccessEditorial

Acknowledgement to Reviewers of JCM in 2019

by JCM Editorial Office

J. Clin. Med. 2020, 9(1), 287; https://doi.org/10.3390/jcm9010287 - 20 Jan 2020

Viewed by 9475

Abstract

Rigorous peer-review is the corner-stone of high-quality academic publishing [...] Full article

34 pages, 887 KiB

Open AccessReview

Genetic and Epigenetic Biomarkers of Immune Checkpoint Blockade Response

by Qingyang Xiao, André Nobre, Pilar Piñeiro, Miguel-Ángel Berciano-Guerrero, Emilio Alba, Manuel Cobo, Volker M. Lauschke and Isabel Barragán

J. Clin. Med. 2020, 9(1), 286; https://doi.org/10.3390/jcm9010286 - 20 Jan 2020

Cited by 45 | Viewed by 8924

Abstract

Checkpoint inhibitor therapy constitutes a promising cancer treatment strategy that targets the immune checkpoints to re-activate silenced T cell cytotoxicity. In recent pivotal trials, immune checkpoint blockade (ICB) demonstrated durable responses and acceptable toxicity, resulting in the regulatory approval of 8 checkpoint inhibitors [...] Read more.

Checkpoint inhibitor therapy constitutes a promising cancer treatment strategy that targets the immune checkpoints to re-activate silenced T cell cytotoxicity. In recent pivotal trials, immune checkpoint blockade (ICB) demonstrated durable responses and acceptable toxicity, resulting in the regulatory approval of 8 checkpoint inhibitors to date for 15 cancer indications. However, up to ~85% of patients present with innate or acquired resistance to ICB, limiting its clinical utility. Current response biomarker candidates, including DNA mutation and neoantigen load, immune profiles, as well as programmed death-ligand 1 (PD-L1) expression, are only weak predictors of ICB response. Thus, identification of novel, more predictive biomarkers that could identify patients who would benefit from ICB constitutes one of the most important areas of immunotherapy research. Aberrant DNA methylation (5mC) and hydroxymethylation (5hmC) were discovered in multiple cancers, and dynamic changes of the epigenomic landscape have been identified during T cell differentiation and activation. While their role in cancer immunosuppression remains to be elucidated, recent evidence suggests that 5mC and 5hmC may serve as prognostic and predictive biomarkers of ICB-sensitive cancers. In this review, we describe the role of epigenetic phenomena in tumor immunoediting and other immune evasion related processes, provide a comprehensive update of the current status of ICB-response biomarkers, and highlight promising epigenomic biomarker candidates. Full article

(This article belongs to the Special Issue Advances and Challenges in Pharmacogenomics)

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Representative mechanisms of resistance to anti-PD-1 immune checkpoint blockade. (A) A low epitope load in the tumor cells normally drives to a minimal immune reinvigoration due to a lower capacity of antigen presenting cells (APC) to present antigen to T cells (low priming) and a lower cytotoxic T cell recognition of the tumor cell antigen. (B) Loss-of-function mutations of Janus kinase (JAKs) desensitize the T cells to the IFN-γ exposure and dramatically decrease the expression level of PD-L1 through lack of activation of the transcription factor STAT. This decrease in PD-L1 leads to both primary and acquired resistance of PD-1 blockade therapy, given that the reinvigoration capacity of T cell through reactivation of the PD-1/PD-L1 axis is abrogated. (C) Deleterious mutations in the gene encoding β2 microglobulin (an MHC class I subunit) lead to loss of antigen presentation, producing resistance to anti-PD-1 drugs. (D) The propensity of the tumor-related PD-1 macrophages to take up anti-PD-1 monoclonal antibodies causes the capture of the anti-PD-1 antibody even from the surface of the PD-1+CD8+ T cells that already bound the drug. This impedes or reverts the anti-PD-1/PD-1 interaction at the cytotoxic T cell provoking resistance to the treatment. (E) In the “escape” phase of the tumor immunoediting, when the tumor is clinically manifested, tolerogenic dendritic cells, myeloid-derived suppressor cells (MDSCs), and tumor-associated macrophages secrete indoleamine-2,3-dioxygenase (IDO), which decreases tryptophan and increases kynurenine. These molecules inhibit effector T cells and NK functions and stimulate regulatory T cells, provoking immunosuppression and enhancing the tolerogenicity of macrophages and dendritic cells. IDO1 also enhances the expansion and activation of MDSCs. All previous alterations suppress the activity of anti-tumor effector T cells. Full article ">

15 pages, 2417 KiB

Open AccessArticle

Performance of FRAX in Predicting Fractures in US Postmenopausal Women with Varied Race and Genetic Profiles

by Qing Wu, Xiangxue Xiao and Yingke Xu

J. Clin. Med. 2020, 9(1), 285; https://doi.org/10.3390/jcm9010285 - 20 Jan 2020

Cited by 12 | Viewed by 4771

Abstract

Background: Whether the Fracture Risk Assessment Tool (FRAX) performed differently in estimating the 10-year fracture probability in women of different genetic profiling and race remained unclear. Methods: The genomic data in the Women’s Health Initiative (WHI) study was analyzed (n = 23,981). [...] Read more.

Background: Whether the Fracture Risk Assessment Tool (FRAX) performed differently in estimating the 10-year fracture probability in women of different genetic profiling and race remained unclear. Methods: The genomic data in the Women’s Health Initiative (WHI) study was analyzed (n = 23,981). The genetic risk score (GRS) was calculated from 14 fracture-associated single nucleotide polymorphisms (SNPs) for each participant. FRAX without bone mineral density (BMD) was used to estimate fracture probability. Results: FRAX significantly overestimated the risk of major osteoporotic fracture (MOF) in the WHI study. The most significant overestimation was observed in women with low GRS (predicted/observed ratio (POR): 1.61, 95% CI: 1.45–1.79) specifically Asian women (POR: 3.5, 95% CI 2.48–4.81) and in African American women (POR: 2.59, 95% CI: 2.33–2.87). Compared to the low GRS group, the 10-year probability of MOF adjusted for the FRAX score was 21% and 30% higher in the median GRS group and high GRS group, respectively. Asian, African American, and Hispanic women respectively had a 78%, 76%, and 56% lower hazard than Caucasian women after the FRAX score was adjusted. The results were similar for hip fractures. Conclusions: Our study suggested the FRAX performance varies significantly by both genetic profile and race in postmenopausal women. Full article

(This article belongs to the Section Orthopedics)

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16 pages, 3837 KiB

Open AccessArticle

Porphyromonas Gingivalis Load is Balanced by 0.20% Chlorhexidine Gel. A Randomized, Double-Blind, Controlled, Microbiological and Immunohistochemical Human Study

by Simonetta D’Ercole, Gianmaria D’Addazio, Silvia Di Lodovico, Tonino Traini, Mara Di Giulio and Bruna Sinjari

J. Clin. Med. 2020, 9(1), 284; https://doi.org/10.3390/jcm9010284 - 20 Jan 2020

Cited by 29 | Viewed by 3786

Abstract

Microbial contamination could compromise the stability of dental implants increasing the risk of inflammatory reactions in the surrounding soft tissues. In this human, randomized, double-blind, clinical study, the presence of Porphyromonas gingivalis on the healing abutment and the inflammatory infiltrate surrounding peri-implant soft [...] Read more.

Microbial contamination could compromise the stability of dental implants increasing the risk of inflammatory reactions in the surrounding soft tissues. In this human, randomized, double-blind, clinical study, the presence of Porphyromonas gingivalis on the healing abutment and the inflammatory infiltrate surrounding peri-implant soft tissues were investigated. Experiments were done in order to clarify the effect of 0.20% chlorhexidine (CHX) versus placebo, applied during each rehabilitation stage. Thirty patients (15 per group) were included. The load of adhering P. gingivalis on the healing screw were quantified by quantitative Polymerase Chain Reaction (qPCR) Taq-Man. Immunohistochemical analysis was carried out on the gingival biopsy. Moreover, clinical data were recorded. Analysis of variance and the Holm–Sidak test was used to evaluate differences between groups. The results showed a significant low presence of P. gingivalis load in healing abutments belonging to the 0.20% CHX group. Overall, the differences in terms of P. gingivalis DNA copy number between two groups were statistically significant (p < 0.01). All implants showed very low plaque and bleeding scores, but the placebo group appeared to have the highest expression of inflammation markers for T Lymphocytes, B Lymphocytes and macrophages Cluster definitions (CD3, CD20 and CD68). The use of 0.20% CHX could be recommended in all clinical procedures as it reduces significantly P. gingivalis load and host inflammatory response around implants. Full article

(This article belongs to the Special Issue Biomaterials and Implants in Development Osteointegration)

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CONSORT 2010 flow diagram. Flow diagram of the progress through the phases of a parallel randomized trial of two groups. Full article ">Figure 2
Explanatory images of the treatment performed (A–D). (A) residual crest. (B) gel inserted into the fixture (T0). (C) soft tissue biopsy (T1). (D) healing abutment placed (T1). Full article ">Figure 3
Detail of a complete case performed in this study: from implant placement to the delivery of permanent restoration. (A–P). (A) Residual crest before incision; (B) bone explosion with periodontal probe to underline the residual bone; (C) implant just positioned; (D) gel inserted into the fixture (T0) ; (E) insertion of cover screw; (F) suture; (G) soft tissue biopsy; (H) gel inserted into the fixture after the removal of cover screw; (I) healing abutment placed; (J) soft tissue before the impression; during this stage the healing abutment was collected for the microbiological analysis (T2) ; (K) impression; (L) gel inserted before the positioning of abutment; (M) definitive abutment location; (N) provisional crown; (O) probing around implant; (P) definitive ceramic restoration delivery. Full article ">Figure 4
Radiographs from the two groups. In the upper images patient from Group A (control group) (clinical images are shown in <a href="#jcm-09-00284-f003" class="html-fig">Figure 3</a>). In the lower images patient from Group B (test group). Pre (Image before the implant placement); T0 (after implant insertion); T1 (second surgical stage at 8 weeks); T2 (before the implant impression at 10 weeks); T3 (restoration’s delivery); T4 (1 year of follow-up). Full article ">Figure 5
Quantification of P. gingivalis on healing abutment by TaqMan Polymerase Chain Reaction (PCR). The differences in terms of colony forming units (CFU) between two groups were statistically significant as reported by * in the picture. Full article ">Figure 6
Immunohistochemical expression at T0, and T1 in group A and B (second surgical stage). Each of these images are in duplicates as one represent the immunohistochemical stain (strep-ABC 20× magnification) meanwhile the duplicate represents the image software analysis (and software analysis 20× magnification). In the first column CD3 expression (strep-ABC 20× magnification and software analysis 20× magnification). In the second column CD20 expression (strep-ABC 20× magnification and software analysis 20× magnification) are shown. In the third column CD68 expression (strep-ABC 20× magnification and software analysis 20× magnification) are shown. In the fourth column CD31 expression (strep-ABC 20× magnification and software analysis 20× magnification). Full article ">

13 pages, 3248 KiB

Open AccessArticle

Differential Diagnosis of Malignant Lymphadenopathy Using Flow Cytometry on Fine Needle Aspirate: Report on 269 Cases

by Carla Griesel, Minodora Desmirean, Tonya Esterhuizen, Sergiu Pasca, Bobe Petrushev, Cristina Selicean, Andrei Roman, Bogdan Fetica, Patric Teodorescu, Carmen Swanepoel, Ciprian Tomuleasa and Ravnit Grewal

J. Clin. Med. 2020, 9(1), 283; https://doi.org/10.3390/jcm9010283 - 20 Jan 2020

Cited by 5 | Viewed by 4189

Abstract

Introduction: Fine needle aspiration (FNA) is frequently the first noninvasive test used for the diagnostic workup of lymphadenopathy. There have been many studies showing its usefulness, especially in conjunction with other techniques for the diagnosis of lymphoma, but it remains inferior to histological [...] Read more.

Introduction: Fine needle aspiration (FNA) is frequently the first noninvasive test used for the diagnostic workup of lymphadenopathy. There have been many studies showing its usefulness, especially in conjunction with other techniques for the diagnosis of lymphoma, but it remains inferior to histological examination. The data regarding this subject have mostly been reported mostly from first-world countries, but are scarce for emerging economies. Thus, the current study assesses the agreement between fine needle aspiration flow cytometry (FNA FC) and histology in the aforementioned region. Material and Methods: We conducted a retrospective study including the FNA FC adenopathy diagnoses made between January 2011 and December 2016 at the Tygerberg Hospital, Cape Town, South Africa. Additional variables included were the histological diagnosis, sex and age of the included patients. Results: In the descriptive part of the current study, 269 FNA FC samples were included. The most frequent diagnoses made on these were represented by B-cell lymphoma, reactive adenopathy, no abnormality detected (NAD), and non-hematological malignancy. In the analytical part of the current study, there were 115 cases included that had both valid FNA FC and histological diagnoses. It could be observed that FNA FC can correctly diagnose B-cell lymphoma in most cases, but it is a poor diagnostic tool especially for Hodgkin lymphoma in this setting as only a four-color flow cytometer was available for diagnosis. Moreover, FNA FC diagnosis of reactive adenopathy and of no abnormalities detected was shown to frequently hide a malignant disease. Conclusion: In countries with scarce resources, FNA FC represents a useful diagnostic tool in the case of B-cell lymphoma, but may misdiagnose reactive adenopathy. Thus, FNA FC should be used in a case-specific manner, in addition to as a screening tool, with the knowledge that in cases with a high clinical suspicion of lymphoma, histological diagnosis is a necessity. Full article

(This article belongs to the Section Hematology)

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Bar plot representing the absolute number of patients with each fine needle aspiration flow cytometry (FNA FC) diagnosis (grey fill) and how many of these cases subsequently had a histological evaluation (red outline). NAD: no abnormalities detected; CLL: chronic lymphocytic leukemia; PCD: plasma cell dyscrasia; NK cell lymphoma: natural killer cell lymphoma; TND: test not done; Non-hematological: non-hematological malignancy; Reactive: reactive inflammatory infiltrate. Full article ">Figure 2
Chord diagram representing the agreement between FNA FC diagnoses (represented with FNA FC diagnoses with FC before their name) and histological diagnoses (represented with histological diagnoses). NAD: no abnormalities detected; CLL: chronic lymphocytic leukemia; PCD: plasma cell dyscrasia; NK cell lymphoma: natural killer cell lymphoma. Full article ">Figure 3
ROC curves and AUC assessing the agreement between FNA FC and histology. NAD: no abnormalities detected; PCD: plasma cell dyscrasia; NK cell lymphoma: natural killer cell lymphoma; Non-hematological: non-hematological malignancy. ROC: receiver operating characteristics. AUC: Area under the Curve. Full article ">Figure 4
Patient’s age and sex distribution between different histological diagnoses. PCD: plasma cell dyscrasia; NK cell lymphoma: natural killer cell lymphoma; Non-hematological: non-hematological malignancy. CLL – chronic lymphocytic leukemia. NAD – No abnormality detected. Full article ">Figure 5
(A–L). Dot blot-based gating strategy for a proof-of-concept study of small lymphocytic leukemia. Full article ">Figure 5 Cont.
(A–L). Dot blot-based gating strategy for a proof-of-concept study of small lymphocytic leukemia. Full article ">

30 pages, 1588 KiB

Open AccessReview

Genetics of Degenerative Cervical Myelopathy: A Systematic Review and Meta-Analysis of Candidate Gene Studies

by Daniel H. Pope, Benjamin M. Davies, Oliver D. Mowforth, A. Ramsay Bowden and Mark R. N. Kotter

J. Clin. Med. 2020, 9(1), 282; https://doi.org/10.3390/jcm9010282 - 20 Jan 2020

Cited by 27 | Viewed by 3751

Abstract

Degenerative cervical myelopathy (DCM) is estimated to be the most common cause of adult spinal cord impairment. Evidence that is suggestive of a genetic basis to DCM has been increasing over the last decade. A systematic search was conducted in MEDLINE, EMBASE, Cochrane, [...] Read more.

Degenerative cervical myelopathy (DCM) is estimated to be the most common cause of adult spinal cord impairment. Evidence that is suggestive of a genetic basis to DCM has been increasing over the last decade. A systematic search was conducted in MEDLINE, EMBASE, Cochrane, and HuGENet databases from their origin up to 14th December 2019 to evaluate the role of single genes in DCM in its onset, clinical phenotype, and response to surgical intervention. The initial search yielded 914 articles, with 39 articles being identified as eligible after screening. We distinguish between those contributing to spinal column deterioration and those contributing to spinal cord deterioration in assessing the evidence of genetic contributions to DCM. Evidence regarding a total of 28 candidate genes was identified. Of these, 22 were found to have an effect on the radiological onset of spinal column disease, while 12 genes had an effect on clinical onset of spinal cord disease. Polymorphisms of eight genes were found to have an effect on the radiological severity of DCM, while three genes had an effect on clinical severity. Polymorphisms of six genes were found to have an effect on clinical response to surgery in spinal cord disease. There are clear genetic effects on the development of spinal pathology, the central nervous system (CNS) response to bony pathology, the severity of both bony and cord pathology, and the subsequent response to surgical intervention. Work to disentangle the mechanisms by which the genes that are reviewed here exert their effects, as well as improved quality of evidence across diverse populations is required for further investigating the genetic contribution to DCM. Full article

(This article belongs to the Special Issue Degenerative Cervical Myelopathy and the Aging Spine)

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16 pages, 3261 KiB

Open AccessArticle

Serum Exosomal MicroRNA, miR-10b-5p, as a Potential Diagnostic Biomarker for Early-Stage Hepatocellular Carcinoma

by Hyo Jung Cho, Jung Woo Eun, Geum Ok Baek, Chul Won Seo, Hye Ri Ahn, Soon Sun Kim, Sung Won Cho and Jae Youn Cheong

J. Clin. Med. 2020, 9(1), 281; https://doi.org/10.3390/jcm9010281 - 20 Jan 2020

Cited by 88 | Viewed by 5771

Abstract

Exosomal microRNAs (exo-miRs) have been promising cancer biomarkers. MiRs in hepatocellular carcinoma (HCC) cell-derived exosomes (HEX) were analyzed to identify reliable serum biomarkers for HCC. To detect overexpressed miRs in HEX, extracted exosomal small RNAs from human HCC cell lines and normal hepatocytes [...] Read more.

Exosomal microRNAs (exo-miRs) have been promising cancer biomarkers. MiRs in hepatocellular carcinoma (HCC) cell-derived exosomes (HEX) were analyzed to identify reliable serum biomarkers for HCC. To detect overexpressed miRs in HEX, extracted exosomal small RNAs from human HCC cell lines and normal hepatocytes were sequenced and analyzed. Clinical significance of the overexpressed miRs in HEX was evaluated using quantitative real-time PCR (qRT-PCR) on serum samples of a validation cohort consisting of 28 healthy individuals, 60 with chronic liver disease, and 90 with HCC. We found 49 significantly overexpressed miRs in HEX compared to a normal hepatocyte. Among them, miR-10b-5p, miR-18a-5p, miR-215-5p, and miR-940 were overexpressed in HCC tissues and also associated with prognosis of HCC in the analysis of a public omics database. qRT-PCR analysis of the four serum exo-miRs in the validation cohort revealed serum exo-miR-10b-5p as a promising biomarker for early-stage HCC with 0.934 area under the curve (AUC) (sensitivity, 90.7%; specificity, 75.0%; cutoff value, 1.8-fold). Overexpression of serum exo-miR-215-5p was found to be significantly associated with poor disease-free survival in patients with HCC. Serum exo-miR-10b-5p is a potential biomarker for early-stage HCC, while serum exo-miR-215-5p can be used as prognostic biomarker for HCC. Full article

(This article belongs to the Section Gastroenterology & Hepatopancreatobiliary Medicine)

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Exosomal microRNAs (exo-miR). Sequencing and detecting specifically overexpressed exo-miRs in hepatocellular carcinoma (HCC). (a) Transmission electron microscopy of the exosomes with 10 nm gold-conjugated anti-CD63 antibody. (b) Pipeline showing analysis of exo-miRs. (c) Venn diagram showing predominantly overexpressed exo-miRs in HCC cell lines compared to expression in THLE-2 cells. (d) Heat map depicting 49 predominantly overexpressed exo-miRs in HCC cell lines. Forty-nine miRs seen to be significantly overexpressed in exosomes from Hep3B as well as Huh-7 cell lines; however, not in those of THLE-2 cells. (e) Ingenuity pathway analysis of 49 overexpressed exo-miRs of HCC cell lines. Full article ">Figure 2
The Cancer Genomic Atlas (TCGA) database analysis, sequencing data of 363 HCC and 50 nontumorous tissues, and corresponding clinical data. (a) Venn diagram showing predominantly overexpressed miRs in HCC cell-derived exosomes and HCC tissues of TCGA dataset. (b) Comparative analysis of substantial expression of the four miRs including miR-10b-5p, miR-18a-5p, miR-215-5p, and miR-940 between nontumorous tissues and HCC tissues. (c) Kaplan–Meier analysis of overall survival based on tissue expressing the four miRs in TCGA database. Full article ">Figure 3
Diagnostic efficiency of serum exo-miRs (miR-10b-5p, miR-18a-5p, miR-215-5p, and miR-940) in diagnosing HCC in test cohort comprising 18 healthy individuals and 19 patients with mUICC stage IV HCC. (a) Comparison of expression of the four serum exo-miRs between normal healthy individuals and patients with HCC. (b) Area under the curve (AUC) and receiver operating characteristics (ROC) of the four serum exo-miRs in diagnosing HCC. Full article ">Figure 4
Diagnostic significance of serum alpha-fetoprotein (AFP), serum exo-miR-10b-5p, and miR-215-5p in diagnosing HCC in the validation cohort comprising 28 normal individuals, 27 patients with chronic hepatitis B, 33 patients with liver cirrhosis, and 90 patients with HCC. (a) Comparison of serum AFP level and expression of two serum exo-miRs based on stage of liver disease. (b) AUC of serum AFP and two serum exo-miRs in diagnosing HCC. (c) AUC of serum AFP and two serum exo-miRs in diagnosing mUICC stage I or II HCC. (d) AUC of serum AFP and two serum exo-miRs in diagnosing mUICC stage I HCC. Full article ">Figure 5
Comparison of diagnostic efficiency of serum AFP, serum exo-miR-10b-5p, and the combination of the serum exo-miR-10b-5p and serum AFP. (a) AUC of the serum markers in diagnosing HCC. From right to left: Diagnosing all stages of HCC, mUICC stage I or II, and mUICC stage I HCC, respectively. (b) AUC of the serum markers in diagnosing HCC in the subgroup composed of CHB, LC, and HCC. From right to left: Diagnosing all stages of HCC, mUICC stage I or II, and mUICC stage I HCC, respectively. Full article ">Figure 6
Prognostic significance of serum exo-miR-10b-5p and exo-miR-215-5p in patients with HCC. (a) Disease-free survival (DFS) and overall survival analysis based on the expression of the two serum exo-miRs in the validation cohort. (b) Expression of the two serum exo-miRs according to the modified Union for International Cancer Control (mUICC) stage guidelines. (c) Expression of the two serum exo-miRs based on vascular invasion. Full article ">

15 pages, 248 KiB

Open AccessArticle

Pain Catastrophising Predicts Alcohol Hangover Severity and Symptoms

by Sam Royle, Lauren Owen, David Roberts and Lynne Marrow

J. Clin. Med. 2020, 9(1), 280; https://doi.org/10.3390/jcm9010280 - 20 Jan 2020

Cited by 9 | Viewed by 3686

Abstract

Alcohol hangover is a cause of considerable social and economic burden. Identification of predictors of alcohol hangover severity have the potential to contribute to reductions in costs associated with both absenteeism/presenteeism and health care. Pain catastrophising (PC) is the tendency to ruminate and [...] Read more.

Alcohol hangover is a cause of considerable social and economic burden. Identification of predictors of alcohol hangover severity have the potential to contribute to reductions in costs associated with both absenteeism/presenteeism and health care. Pain catastrophising (PC) is the tendency to ruminate and describe a pain experience in more exaggerated terms. The current study examines the possibility that this cognitive coping strategy may influence experience of alcohol hangover. The aims of the current study were to (1) examine the relationship between hangover severity and PC, (2) explore and identify discreet factors within the Acute Hangover Scale (AHS) and (3) explore whether independent factors/dimensions of acute hangover are differentially predicted by PC. A retrospective survey (n = 86) was conducted in which participants completed the Acute Hangover Scale (AHS); the Pain Catastrophising Scale (PCS); a questionnaire pertaining to the amount of alcohol consumed; and a demographic information questionnaire. Regression analyses showed a significant relationship between PC and hangover severity scores and demonstrated that PC was, in fact, a stronger predictor of perceived hangover severity than estimated peak blood alcohol concentrations (eBACs). Factor analysis of the AHS scale, resulted in the identification of two distinct symptom dimensions; ‘Headache and thirst’, and ‘Gastric and cardiovascular’ symptoms. Regression analyses showed that both eBAC and PCS score were significantly associated with ‘Headache and thirst’. However, only PCS score was associated with ‘Gastric and cardiovascular’ symptoms. These novel findings implicate a role for cognitive coping strategies in self-reports of alcohol hangover severity, and may have implications for understanding behavioural response to hangover, as well as suggesting that hangover and PC may be important factors mediating the motivation to drink and/or abuse alcohol, with potential implications in addiction research. Furthermore, these findings suggest that distinct alcohol hangover symptoms may be associated with different mechanisms underlying the experience of alcohol hangover. Full article

(This article belongs to the Special Issue The Alcohol Hangover: Causes, Consequences, and Treatment)

15 pages, 2131 KiB

Open AccessArticle

Bone Marrow-Derived Mesenchymal Stromal Cells (MSCs) Modulate the Inflammatory Character of Alveolar Macrophages from Sarcoidosis Patients

by Ian McClain Caldwell, Christopher Hogden, Krisztian Nemeth, Michael Boyajian, Miklos Krepuska, Gergely Szombath, Sandra MacDonald, Mehrnoosh Abshari, Joel Moss, Lynn Vitale-Cross, Joseph R Fontana and Eva Mezey

J. Clin. Med. 2020, 9(1), 278; https://doi.org/10.3390/jcm9010278 - 19 Jan 2020

Cited by 14 | Viewed by 3850

Abstract

Sarcoidosis is a devastating inflammatory disease affecting many organs, especially the lungs and lymph nodes. Bone marrow-derived mesenchymal stromal cells (MSCs) can “reprogram” various types of macrophages towards an anti-inflammatory phenotype. We wanted to determine whether alveolar macrophages from sarcoidosis subjects behave similarly [...] Read more.

Sarcoidosis is a devastating inflammatory disease affecting many organs, especially the lungs and lymph nodes. Bone marrow-derived mesenchymal stromal cells (MSCs) can “reprogram” various types of macrophages towards an anti-inflammatory phenotype. We wanted to determine whether alveolar macrophages from sarcoidosis subjects behave similarly by mounting an anti-inflammatory response when co-cultured with MSCs. Fifteen sarcoidosis and eight control subjects underwent bronchoscopy and bronchoalveolar lavage (BAL). Unselected BAL cells (70–94% macrophages) were isolated and cultured with and without MSCs from healthy adults. Following stimulation of the cultured cells with lipopolysaccharide, the medium was removed to measure interleukin 10 and tumor necrosis factor alpha (IL-10 and TNF-α). In two additional sarcoidosis subjects, flow cytometry was used to study intracellular cytokines and surface markers associated with alveolar macrophages to confirm the results. Unselected BAL cells from sarcoidosis subjects co-cultured with MSCs showed a reduction in TNF-α (pro-inflammatory M1) and an increase in IL-10 (anti-inflammatory M2) in 9 of 11 samples studied. Control subject samples showed few, if any, differences in cytokine production. Unselected BAL cells from two additional patients analyzed by flow cytometry confirmed a switch towards an anti-inflammatory state (i.e., M1 to M2) after co-culture with MSCs. These results suggest that, similarly to other macrophages, alveolar macrophages also respond to MSC contacts by changing towards an anti-inflammatory phenotype. Based on our results, we hypothesize that mesenchymal stromal cells applied to the airways might alleviate lung inflammation and decrease steroid need in patients with sarcoidosis. Full article

(This article belongs to the Special Issue Management of Sarcoidosis: Challenges and Solutions)

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Figure 1
Percent change in cytokine production of bone marrow stromal cell (MSC) and bronchoalveolar lavage (BAL) cell co-cultures stimulated with lipopolysaccharide (LPS). (A) Cytokine (IL-10—left half and TNF-α—right half) concentrations were measured from tissue culture medium of LPS-stimulated co-cultures of MSCs and BAL cells from sarcoidosis (red) and control (blue) subjects. Of the 11 cases presented, 2 sarcoidosis samples (19 and 25) diverged from the trend in one cytokine or the other (indicated by arrows and asterisks). Control subject samples showed few, if any, differences in cytokine production with no obvious trend. (B) Average percent changes in anti-inflammatory (IL-10) and the pro-inflammatory (TNF-α) cytokines in the co-culture medium are shown for both groups. The graph shows that all of the samples from the sarcoidosis subjects shift towards an anti-inflammatory state (increase in IL-10 and decrease in TNF-α), while samples from the control subjects do not (n = 9 for sarcoidosis subjects, n = 7 for control subjects, unpaired student’s t test; data are shown as mean ± SEM; p = 0.011 for IL-10 and 0.029 for TNF-α). Full article ">Figure 2
Flow cytometry analysis of mononuclear cells (90% of which were macrophages) freshly prepared from a sarcoidosis subject (sarcoidosis subject 16). Bronchoalveolar lavage (BAL) cells were plated and cultured for 16 h with (B,D,F) and without (A,C,E) the presence of bone marrow stromal cells (MSCs). Surface marker CD206 was used to identify alveolar macrophages (AMs), and intracellular flow cytometry was performed to determine changes in their cytokine production. Co-cultured BAL cells increased their IL-10 (A,B) and decreased their TNF-α (C,D) production. CD163 (a surface marker thought to indicate an anti-inflammatory state) also increased (E,F). These changes suggest a shift from a pro-inflammatory towards an anti-inflammatory state of AMs following contact with MSCs. Full article ">Figure 3
Intracellular cytokine ratios of alveolar macrophages (AMs) suggest a change from M1 (pro-inflammatory) to M2 (anti-inflammatory) status after co-culture with bone marrow stromal cells (MSCs). (A) Bronchoalveolar lavage (BAL) cells are depicted in red, BAL cells from co-cultures are pictured in blue. The co-cultured BAL cells demonstrate a clear shift towards the right, indicating an increase in the mean fluorescent intensity (MFI) of the IL-10+ population, and upwards, indicating an increase in the MFI of CD206 in these cells. Due to the lower percentage of cells producing TNF-α, a leftward shift is not as evident, although an upward shift indicating increased MFI of CD206 in these cells is present. (B) The fold change of the ratio of IL-10 to TNF-α for the BAL–MSC co-culture is shown for both subjects (sarcoidosis subjects 16 and 17). BAL cells cultured alone are depicted by the red bars (the ratio was converted to a value of 1), and co-cultured BAL cells are depicted by the blue bars (representing the fold increase of the ratio). The bar graph shows that—although not to the same extent—the BAL cells shifted towards a more anti-inflammatory state in both subjects. Full article ">Figure 4
Flow cytometry analysis of mononuclear cells (73% of which were macrophages) freshly prepared from a sarcoidosis subject (sarcoidosis subject 17). Bronchoalveolar lavage (BAL) cells were plated and cultured for 16 hours with (B,D,F) and without (A,C,E) the presence of bone marrow stromal cells (MSCs). Surface marker CD206 was used to identify alveolar macrophages (AMs), and intracellular flow cytometry was performed to determine changes in their cytokine production. Co-cultured BAL cells almost tripled their IL-10 (A,B) while their TNF-α (C,D) production barely changed. Interestingly, CD163 (a surface marker thought to indicate an anti-inflammatory state) decreased (E,F). Overall, these changes may suggest a shift from a pro-inflammatory towards an anti-inflammatory state of AMs following contact with MSCs. Full article ">

10 pages, 647 KiB

Open AccessArticle

Safety of Abatacept in Italian Patients with Rheumatoid Arthritis and Interstitial Lung Disease: A Multicenter Retrospective Study

by Giulia Cassone, Andreina Manfredi, Fabiola Atzeni, Vincenzo Venerito, Caterina Vacchi, Valentina Picerno, Federica Furini, Gian Luca Erre, Paola Tomietto, Anna Laura Fedele, Giovanni Della Casa, Valeria Nucera, Chiara Giannitti, Carlo Salvarani and Marco Sebastiani

J. Clin. Med. 2020, 9(1), 277; https://doi.org/10.3390/jcm9010277 - 19 Jan 2020

Cited by 50 | Viewed by 4995

Abstract

Background: Treatment of rheumatoid arthritis (RA)-related interstitial lung disease (ILD) is challenging, and many conventional and biologic disease-modifying anti-rheumatic drugs (DMARDs) have been associated with ILD development or progression. The aim of this multicentric retrospective study was to analyze the evolution of ILD [...] Read more.

Background: Treatment of rheumatoid arthritis (RA)-related interstitial lung disease (ILD) is challenging, and many conventional and biologic disease-modifying anti-rheumatic drugs (DMARDs) have been associated with ILD development or progression. The aim of this multicentric retrospective study was to analyze the evolution of ILD in Italian RA-ILD patients treated with abatacept (ABA). Methods: All RA-ILD patients treated with ABA for at least six months were retrospectively evaluated. Serology, previous and concurrent therapies, chest high-resolution computer tomography (HRCT), forced vital capacity (FVC), and lung diffusion of carbon monoxide (CO, DLCO) were collected. Results: Forty-four patients were included; HRCT, FVC, and DLCO were analyzed at baseline, at one year, and at the end of follow-up. A remission or a low disease activity of RA was reached in 41/44 patients. Overall, FVC and DLCO remained stable or increased in 86.1% and 91.7% of patients, respectively, while HRCT was stable or improved in 81.4% of them. Previous and concurrent treatments, in particular, methotrexate, serology, age, sex, joint and lung disease duration were not associated with the outcome at univariate analysis. Conclusion: The management of RA-ILD patients remains a critical unmet medical need. Waiting for prospective controlled studies, ABA has shown a good safety profile in our cohort of Italian RA-ILD patients. Full article

(This article belongs to the Special Issue Diagnosis, Clinical Features and Management of Interstitial Lung Diseases in Rheumatic Disorders)

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10 pages, 565 KiB

Open AccessArticle

Expression Patterns of Circulating MicroRNAs in the Risk Stratification of Bicuspid Aortopathy

by Evaldas Girdauskas, Niklas Neumann, Johannes Petersen, Tatiana Sequeira-Gross, Shiho Naito, Maria von Stumm, Yskert von Kodolitsch, Hermann Reichenspurner and Tanja Zeller

J. Clin. Med. 2020, 9(1), 276; https://doi.org/10.3390/jcm9010276 - 19 Jan 2020

Cited by 11 | Viewed by 2649

Abstract

Objective: Aortic size-based criteria are of limited value in the prediction of aortic events, while most aortic events occur in patients with proximal aortic diameters < 50 mm. Serological biomarkers and especially circulating microRNAs (miRNAs) have been proposed as an elegant tool to [...] Read more.

Objective: Aortic size-based criteria are of limited value in the prediction of aortic events, while most aortic events occur in patients with proximal aortic diameters < 50 mm. Serological biomarkers and especially circulating microRNAs (miRNAs) have been proposed as an elegant tool to improve risk stratification in patients with different aortopathies. Therefore, we aimed to evaluate the levels of circulating miRNAs in a surgical cohort of patients presenting with bicuspid aortic valve disease and distinct valvulo-aortic phenotypes. Methods: We prospectively examined a consecutive cohort of 145 patients referred for aortic valve surgery: (1) Sixty three patients (mean age 47 ± 11 years, 92% male) with bicuspid aortic valve regurgitation and root dilatation (BAV-AR), (2) thirty two patients (mean age 59 ± 11 years, 73% male) with bicuspid aortic valve stenosis (BAV-AS), and (3) fifty patients (mean age 56 ± 14 years, 55% male) with tricuspid aortic valve stenosis and normal aortic root diameters (TAV-AS) who underwent aortic valve+/-proximal aortic surgery at a single institution. MicroRNAs analysis included 11 miRNAs, all published previously in association with aortopathies. Endpoints of our study were (1) correlation between circulating miRNAs and aortic diameter and (2) comparison of circulating miRNAs in distinct valvulo-aortic phenotypes. Results: We found a significant inverse linear correlation between circulating miRNAs levels and proximal aortic diameter in the whole study cohort. The strongest correlation was found for miR-17 (r = −0.42, p < 0.001), miR-20a (r = −0.37, p < 0.001), and miR-106a (r = −0.32, p < 0.001). All miRNAs were significantly downregulated in BAV vs. TAV with normal aortic root dimensions Conclusions: Our data demonstrate a significant inverse correlation between circulating miRNAs levels and the maximal aortic diameter in BAV aortopathy. When comparing miRNAs expression patterns in BAV vs. TAV patients with normal aortic root dimensions, BAV patients showed significant downregulation of analyzed miRNAs as compared to their TAV counterparts. Further multicenter studies in larger cohorts are needed to further validate these results. Full article

(This article belongs to the Special Issue Current Challenges in the Diagnosis and Treatment of Hereditary and Congenital Aortopathies)

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21 pages, 795 KiB

Open AccessReview

Epidemiology, Treatment, and Prevention of Nosocomial Bacterial Pneumonia

by Shio-Shin Jean, Yin-Chun Chang, Wei-Cheng Lin, Wen-Sen Lee, Po-Ren Hsueh and Chin-Wan Hsu

J. Clin. Med. 2020, 9(1), 275; https://doi.org/10.3390/jcm9010275 - 19 Jan 2020

Cited by 79 | Viewed by 17207

Abstract

Septicaemia likely results in high case-fatality rates in the present multidrug-resistant (MDR) era. Amongst them are hospital-acquired pneumonia (HAP) and ventilator-associated pneumonia (VAP), two frequent fatal septicaemic entities amongst hospitalised patients. We reviewed the PubMed database to identify the common organisms implicated in [...] Read more.

Septicaemia likely results in high case-fatality rates in the present multidrug-resistant (MDR) era. Amongst them are hospital-acquired pneumonia (HAP) and ventilator-associated pneumonia (VAP), two frequent fatal septicaemic entities amongst hospitalised patients. We reviewed the PubMed database to identify the common organisms implicated in HAP/VAP, to explore the respective risk factors, and to find the appropriate antibiotic choice. Apart from methicillin-resistant Staphylococcus aureus and Pseudomonas aeruginosa, extended-spectrum β-lactamase-producing Enterobacteriaceae spp., MDR or extensively drug-resistant (XDR)-Acinetobacter baumannii complex spp., followed by Stenotrophomonas maltophilia, Chryseobacterium indologenes, and Elizabethkingia meningoseptica are ranked as the top Gram-negative bacteria (GNB) implicated in HAP/VAP. Carbapenem-resistant Enterobacteriaceae notably emerged as an important concern in HAP/VAP. The above-mentioned pathogens have respective risk factors involved in their acquisition. In the present XDR era, tigecycline, colistin, and ceftazidime-avibactam are antibiotics effective against the Klebsiella pneumoniae carbapenemase and oxacillinase producers amongst the Enterobacteriaceae isolates implicated in HAP/VAP. Antibiotic combination regimens are recommended in the treatment of MDR/XDR-P. aeruginosa or A. baumannii complex isolates. Some special patient populations need prolonged courses (>7-day) and/or a combination regimen of antibiotic therapy. Implementation of an antibiotic stewardship policy and the measures recommended by the United States (US) Institute for Healthcare were shown to decrease the incidence rates of HAP/VAP substantially. Full article

(This article belongs to the Special Issue Diagnosis and Treatment of Pneumonia)

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20 pages, 2287 KiB

Open AccessArticle

Comparative Genomic Mapping Implicates LRRK2 for Intellectual Disability and Autism at 12q12, and HDHD1, as Well as PNPLA4, for X-Linked Intellectual Disability at Xp22.31

by Jonathan D. J. Labonne, Terri M. Driessen, Marvin E. Harris, Il-Keun Kong, Soumia Brakta, John Theisen, Modibo Sangare, Lawrence C. Layman, Cheol-Hee Kim, Janghoo Lim and Hyung-Goo Kim

J. Clin. Med. 2020, 9(1), 274; https://doi.org/10.3390/jcm9010274 - 19 Jan 2020

Cited by 14 | Viewed by 6507

Abstract

We report a genomic and phenotypic delineation for two chromosome regions with candidate genes for syndromic intellectual disability at 12q12 and Xp22.31, segregating independently in one family with four affected members. Fine mapping of three affected members, along with six unreported small informative [...] Read more.

We report a genomic and phenotypic delineation for two chromosome regions with candidate genes for syndromic intellectual disability at 12q12 and Xp22.31, segregating independently in one family with four affected members. Fine mapping of three affected members, along with six unreported small informative CNVs, narrowed down the candidate chromosomal interval to one gene LRRK2 at 12q12. Expression studies revealed high levels of LRRK2 transcripts in the whole human brain, cerebral cortex and hippocampus. RT-qPCR assays revealed that LRRK2 transcripts were dramatically reduced in our microdeletion patient DGDP289A compared to his healthy grandfather with no deletion. The decreased expression of LRRK2 may affect protein–protein interactions between LRRK2 and its binding partners, of which eight have previously been linked to intellectual disability. These findings corroborate with a role for LRRK2 in cognitive development, and, thus, we propose that intellectual disability and autism, displayed in the 12q12 microdeletions, are likely caused by LRRK2. Using another affected member, DGDP289B, with a microdeletion at Xp22.31, in this family, we performed the genomic and clinical delineation with six published and nine unreported cases. We propose HDHD1 and PNPLA4 for X-linked intellectual disability in this region, since their high transcript levels in the human brain substantiate their role in intellectual functioning. Full article

(This article belongs to the Special Issue Genetics of Autism Spectrum Disorders)

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23 pages, 733 KiB

Open AccessReview

Mixed Neuroendocrine Non-Neuroendocrine Neoplasms: A Systematic Review of a Controversial and Underestimated Diagnosis

by Melissa Frizziero, Bipasha Chakrabarty, Bence Nagy, Angela Lamarca, Richard A. Hubner, Juan W. Valle and Mairéad G. McNamara

J. Clin. Med. 2020, 9(1), 273; https://doi.org/10.3390/jcm9010273 - 19 Jan 2020

Cited by 82 | Viewed by 5968

Abstract

Mixed neuroendocrine non-neuroendocrine neoplasms (MiNENs) represent a rare diagnosis of the gastro-entero-pancreatic tract. Evidence from the current literature regarding their epidemiology, biology, and management is of variable quality and conflicting. Based on available data, the MiNEN has an aggressive biological behaviour, mostly driven [...] Read more.

Mixed neuroendocrine non-neuroendocrine neoplasms (MiNENs) represent a rare diagnosis of the gastro-entero-pancreatic tract. Evidence from the current literature regarding their epidemiology, biology, and management is of variable quality and conflicting. Based on available data, the MiNEN has an aggressive biological behaviour, mostly driven by its (often high-grade) neuroendocrine component, and a dismal prognosis. In most cases, the non-neuroendocrine component is of adenocarcinoma histology. Due to limitations in diagnostic methods and poor awareness within the scientific community, the incidence of MiNENs may be underestimated. In the absence of data from clinical trials, MiNENs are commonly treated according to the standard of care for pure neuroendocrine carcinomas or adenocarcinomas from the same sites of origin, based on the assumption of a biological similarity to their pure counterparts. However, little is known about the molecular aberrations of MiNENs, and their pathogenesis remains controversial; molecular/genetic studies conducted so far point towards a common monoclonal origin of the two components. In addition, mutations in tumour-associated genes, including TP53, BRAF, and KRAS, and microsatellite instability have emerged as potential drivers of MiNENs. This systematic review (91 full manuscripts or abstracts in English language) summarises the current reported literature on clinical, pathological, survival, and molecular/genetic data on MiNENs. Full article

(This article belongs to the Special Issue Advances in Diagnosis and Therapy of Neuroendocrine Neoplasms)

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17 pages, 1210 KiB

Open AccessReview

Could IL-17A Be a Novel Therapeutic Target in Diabetic Nephropathy?

by Carolina Lavoz, Sandra Rayego-Mateos, Macarena Orejudo, Lucas Opazo-Ríos, Vanessa Marchant, Laura Marquez-Exposito, Antonio Tejera-Muñoz, Juan F. Navarro-González, Alejandra Droguett, Alberto Ortiz, Jesús Egido, Sergio Mezzano, Raúl R. Rodrigues-Diez and Marta Ruiz-Ortega

J. Clin. Med. 2020, 9(1), 272; https://doi.org/10.3390/jcm9010272 - 19 Jan 2020

Cited by 31 | Viewed by 6364

Abstract

Chronic kidney disease has become a major medical issue in recent years due to its high prevalence worldwide, its association with premature mortality, and its social and economic implications. A number of patients gradually progress to end-stage renal disease (ESRD), requiring then dialysis [...] Read more.

Chronic kidney disease has become a major medical issue in recent years due to its high prevalence worldwide, its association with premature mortality, and its social and economic implications. A number of patients gradually progress to end-stage renal disease (ESRD), requiring then dialysis and kidney transplantation. Currently, approximately 40% of patients with diabetes develop kidney disease, making it the most prevalent cause of ESRD. Thus, more effective therapies for diabetic nephropathy are needed. In preclinical studies of diabetes, anti-inflammatory therapeutic strategies have been used to protect the kidneys. Recent evidence supports that immune cells play an active role in the pathogenesis of diabetic nephropathy. Th17 immune cells and their effector cytokine IL-17A have recently emerged as promising targets in several clinical conditions, including renal diseases. Here, we review current knowledge regarding the involvement of Th17/IL-17A in the genesis of diabetic renal injury, as well as the rationale behind targeting IL-17A as an additional therapy in patients with diabetic nephropathy. Full article

(This article belongs to the Special Issue Diabetic Nephropathy: Diagnosis, Prevention and Treatment)

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Th17 differentiation. Naïve CD4+ T lymphocytes can be differentiated into different T cell subtypes, including Treg or Th17 immune cells. This process is regulated by particular cytokines and activation of specific transcription factors, as indicated. Moreover, mixed phenotypes have also been described. Full article ">Figure 2
Proposed mechanism of IL-17A-induced renal damage in diabetic nephropathy. Under diabetic conditions, renal resident cells are activated and can produce different mediators that could contribute to recruit immune cells into the kidney. Infiltrating Th17 cells can locally produce IL-17A in the diabetic kidney. Then, IL-17A acting on IL-17R on resident renal cells can produce additional proinflammatory mediators, contributing to sustained inflammation. Moreover, IL-17A acting on tubular epithelial cells can induce phenotype changes, such as partial epithelial-to-mesenchymal transition (EMT) and secretome changes. By these mechanisms, IL-17A participates in the amplification of the inflammatory response and the progression of renal damage, finally leading to tubulointerstitial fibrosis. Full article ">Figure 3
Intracellular mechanisms involved in inflammatory responses of IL-17A in the kidney. IL-17A can binds to its receptors and activates several intracellular mechanisms. The activation of NF-κB pathway and the upregulation of proinflammatory factors, such as MCP-1 can contribute to renal inflammation, as proposed under diabetic conditions. IL-17A can also activate other mechanisms, such as protein kinases and redox processes, but their role in renal damage have not been fully demonstrated. Full article ">

11 pages, 2096 KiB

Open AccessArticle

Highly Sensitive Detection of IDH2 Mutations in Acute Myeloid Leukemia

by Jessica Petiti, Valentina Rosso, Eleonora Croce, Vanessa Franceschi, Giacomo Andreani, Matteo Dragani, Marco De Gobbi, Monia Lunghi, Giuseppe Saglio, Carmen Fava, Marco Lo Iacono and Daniela Cilloni

J. Clin. Med. 2020, 9(1), 271; https://doi.org/10.3390/jcm9010271 - 19 Jan 2020

Cited by 11 | Viewed by 3904

Abstract

Background: Acute myeloid leukemia is a heterogeneous hematological disease, characterized by karyotypic and molecular alterations. Mutations in IDH2 have a role in diagnosis and as a minimal residue disease marker. Often the variant allele frequency during follow up is less than 20%, which [...] Read more.

Background: Acute myeloid leukemia is a heterogeneous hematological disease, characterized by karyotypic and molecular alterations. Mutations in IDH2 have a role in diagnosis and as a minimal residue disease marker. Often the variant allele frequency during follow up is less than 20%, which represents the limit of detection of Sanger sequencing. Therefore, the development of sensitive methodologies to identify IDH2 mutations might help to monitor patients’ response to therapy. We compared three different methods to identify and monitor IDH2 mutations in patients’ specimens. Methods: Performances of PNA-PCR clamping, droplet digital PCR and Sanger for IDH2 status identification were evaluated and compared in 96 DNA patients’ specimens. Results: In contrast with Sanger sequencing, our results highlighted the concordance between PNA clamping and digital PCR. Furthermore, PNA-PCR clamping was able to detect more mutated DNA with respect to Sanger sequencing that showed several false negatives independently from the allelic frequency. Conclusions: We found that PNA-PCR clamping and digital PCR identified IDH2 mutations in DNA samples with comparable results in a percentage significantly higher compared to Sanger sequencing. PNA-PCR clamping can be used even in laboratories not equipped for sophisticated analyses, decreasing cost and time for IDH2 characterization. Full article

(This article belongs to the Special Issue Advances in Acute Myeloid Leukemia)

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PNA-PCR clamping experimental design. Amplification of IDH2 was performed in presence of PNA probe (light blue), designed on the WT sequence, and Primer competitor (purple), designed on the mutated sequence. In these conditions, the PCR of the WT sequence is inhibited by the inability of the primer competitor to detach the perfect hybridization PNA/DNA. In contrast, in the presence of IDH2mut, PNA/DNA duplex is highly destabilized by the Primer competitor, allowing a strong amplification of the target sequence. Full article ">Figure 2
Electrophoretic runs of PCR reactions: 10 μL of each amplicon were loaded on 2% Agarose-TBE 1× gel with 5 μg/mL EtBr and run at 120 V for 30 min. PNA-PCR clamping for IDH2 R140Q and R172K carried out in absence (PNA−) or in presence (PNA+) of the PNA probe. (A) PNA-PCR clamping in DNA from AML samples and control plasmid. The results, read in duplicate, were: IDH2 WT, if there was no amplification in (PNA+) and amplification in (PNA−) in both the PCR (for R140Q and R172K reaction); IDH2 R140Q, if there was amplification in both (PNA+) and (PNA−) in R140Q reaction and there was no amplification in (PNA+) and amplification in (PNA−) in R172K reaction; IDH2 R172K, if there was amplification in both (PNA+) and (PNA−) in R172K reaction and there was no amplification in (PNA+) and amplification in (PNA−) in R140Q reaction. (B) PNA-PCR clamping sensitivity assessed mixing at different ratio IDH2mut and IDH2wt template in the same PCR reaction. Dilutions were as follows: 100, 50, 30, 20, 10, 5, 1 and 0.5% IDH2mut (R140Q or R172K) all brought to 100% with the respective amount of IDH2wt template. The percentage of the mutated template is indicated. Red boxes highlight the limit of detection of the PNA-PCR clamping method for each mutation in the study. Full article ">Figure 3
Agreement charts for comparing IDH2 status evaluation techniques. In agreement plots, the black blocks indicate the accordance of results for each method with respect to the ddPCR, selected as a calibrator for the analysis. ddPCR and PNA-PCR clamping (B) had a greater agreement with respect to ddPCR and Sanger sequencing (A), as also suggested by the shorter segment of the red 45° diagonal line not included in the black blocks. The 2 × 2 tables alongside underline the differences of each methodology with respect to ddPCR. Full article ">Figure 4
Comparison of techniques for IDH2 status evaluation in three patients affected by AML. (A). Electrophoretic runs of PNA-PCR clamping carried out in absence (PNA−) or in presence (PNA+) of the PNA probe. The results, read in duplicate, are: IDH2wt (#1), no amplification in (+) and amplification in (−); IDH2mut (#2 and #3), amplification in both (+) and (−). (B). Sequencing chromatograms: red boxes indicate nucleotide affected by IDH2 R140Q mutation. Black peak indicates WT nucleotide (G) in #1 and #3; while double peak black and green (G/A) indicate IDH2 R140Q heterozygous mutation in #2. (C). 2D plots of ddPCR analysis. For each plot, the amplitude in channel 1 (IDH2 R140Q) is represented on the y-axis with the amplitude in channel 2 (IDH2 WT) represented on the x-axis. Four clusters are identified as single-positive for IDH2 mutated (blue) and IDH2 wild type (green), double-positive (orange) and double-negative (grey). The percentages of IDH2 R140Q/IDH2 WT are indicated in each 2D plot. Full article ">Figure 5
IDH2 status evaluation techniques detection limits and results agreement. The graphs indicate all the variant allelic frequency (VAF) of IDH2 mutations identified in AML patients with ddPCR and sorted from 0% (WT) to 50% mutated (heterozygous). ddPCR identified IDH2 mutant already by 0.6% of VAF, while PNA-PCR clamping failed to identify positive patients under the 1% of VAF (zoom of this region is showed in the dotted circle). Contrariwise, the discriminatory ability of Sanger sequence was randomly distributed in all ranges of IDH2 mutation percentage, decreasing the robustness of the analysis. Negative (−), positive (+) and false negative (*). Full article ">

13 pages, 2330 KiB

Open AccessArticle

The Radical Scavenger NZ-419 Suppresses Intestinal Polyp Development in Apc-Mutant Mice

by Yurie Kurokawa, Gen Fujii, Susumu Tomono, Shingo Miyamoto, Takahiro Hamoya, Maiko Takahashi, Takumi Narita, Masami Komiya, Masaki Kobayashi, Yoshikazu Higami and Michihiro Mutoh

J. Clin. Med. 2020, 9(1), 270; https://doi.org/10.3390/jcm9010270 - 18 Jan 2020

Cited by 1 | Viewed by 2891

Abstract

Colorectal cancer is the fourth leading cause of cancer death worldwide, and it is important to establish effective methods for preventing colorectal cancer. One effective prevention strategy could be the use of antioxidants. However, the role of the direct antioxidative function of antioxidants [...] Read more.

Colorectal cancer is the fourth leading cause of cancer death worldwide, and it is important to establish effective methods for preventing colorectal cancer. One effective prevention strategy could be the use of antioxidants. However, the role of the direct antioxidative function of antioxidants against carcinogenesis has not been clarified. Thus, we aimed to determine whether the direct removal of reactive oxygen species by a hydroxyl radical scavenger, NZ-419, could inhibit colorectal carcinogenesis. NZ-419 is a creatinine metabolite that has been shown to be safe and to inhibit the progression of chronic kidney disease in rats, and it is now under clinical development. In the present study, we demonstrated that NZ-419 eliminated reactive oxygen species production in HCT116 cells after H₂O₂ stimulation and suppressed H₂O₂-induced Nrf2 promoter transcriptional activity. The administration of 500 ppm NZ-419 to Apc-mutant Min mice for 8 weeks resulted in a decrease in the number of polyps in the middle segment of the small intestine to 62.4% of the value in the untreated control (p < 0.05 vs. control group). As expected, NZ-419 treatment affected the levels of reactive carbonyl species, which are oxidative stress markers in the serum of Min mice. Suppression of the mRNA levels of the proliferation-associated factor c-Myc was observed in intestinal polyps of Min mice after NZ-419 treatment, with a weak suppression of epithelial cell proliferation assessed by proliferation cell nuclear antigen (PCNA) staining in the intestinal polyps. This study demonstrated that NZ-419 suppress the development of intestinal polyps in Min mice, suggesting the utility of radical scavenger/antioxidants as a cancer chemopreventive agent. Full article

(This article belongs to the Special Issue Molecular Mechanisms of Cancer Chemoprevention)

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NZ-419 eliminates reactive oxygen species (ROS) production in HCT116 cells. (A) Chemical structure of NZ-419. (B) HCT116 cells were treated with 1 mM NZ-419 (NZ), 5 mM N-Acetyl-l-cysteine (NAC), 100 µM 5-aminosalicylic acid (5-ASA) for 30 min and then treated with 0.5 mM H2O2 and further incubated for 10 min to evaluate the amount of ROS production. NAC and 5-ASA are positive controls. Data are presented as the means ± SD (n = 3). Asterisks indicate significant difference from the untreated control group at * p < 0.05, ** p < 0.01, and *** p < 0.005. The data are representative data obtained from more than three independent experiments. Full article ">Figure 2
NZ-419 suppresses H2O2-induced Nrf2 promoter transcriptional activity. HCT116 cells were treated with NZ-419 (10,000 µM) and incubated for 30 min at 37 °C. After incubation, cells were treated with 40 μM H2O2 for 24 h (A), or for 24 h without H2O2. (B) The basal luciferase activity level of the control was set as 1.0. The data are presented as the means ± SD (n = 3). ** p < 0.01, *** p < 0.005 vs. 0 control. The data are representative data obtained from more than three independent experiments. Full article ">Figure 3
Amount of food intake and body weight changed throughout the experimental period. Amount of food intake /mouse/day (A) and body weight of each mouse (B) were measured weekly. Full article ">Figure 4
NZ-419 inhibited the size of intestinal polyps. The size distributions of the intestinal polyps in the basal control diet and NZ-419-treated groups are shown. Full article ">Figure 5
Reactive carbonyl species (RCs) maps plotting free RCs detected in the serum samples of Min mice. All free RCs detected in the serum samples taken from 0 (A) or 1000 (B) ppm NZ-419-treated Min mice are shown. The RCs are plotted as circles as a function of their retention times (horizontal axis) and m/z values (vertical axis). The areas of the circles represent the intensities of the peaks of the RCs relative to that of the internal standard (IS). The RCs abbreviations are listed in the Materials and Methods section. (C) The comparative RCs profiles of the serum samples from 1000 ppm NZ-419-treated Min mice. The closed diamonds indicate that the RCs levels were significantly different between non-treated and NZ-419-treated mice (p < 0.05). Full article ">Figure 6
Suppression of c-Myc and Cdk4 in non-polyp intestinal mucosa or/and polyp parts of Min mice treated or not with 1000 ppm NZ-419. The mice received diets containing NZ-419 at the indicated doses for 8 weeks. Quantitative real-time PCR analyses were performed to determine the c-Myc (A) and Cdk4 (B) mRNA expression levels in the polyps and non-polyp intestinal mucosa of Min mice. The data are normalized to GAPDH and presented as the means ± SD (n = 3). The basal mRNA expression level in the control was set as 1.0. * p < 0.05 vs. 0 ppm. Full article ">Figure 7
NZ-419 inhibited epithelial cell proliferation. Change in the ratio of proliferation cell nuclear antigen (PCNA)-positive cells in the intestinal polyps with NZ-419 treatment. Immunohistochemical staining for PCNA in the intestinal polyps of Min mice was performed in the basal diet group and 1000 ppm NZ-419-containing diet group. Calculations of the ratio of PCNA-positive cells are described in the Materials and Methods section. Bars indicate the SD (n = 7~11). Full article ">

11 pages, 2420 KiB

Open AccessArticle

Split-Liver Ex Situ Machine Perfusion: A Novel Technique for Studying Organ Preservation and Therapeutic Interventions

by Viola Huang, Negin Karimian, Danielle Detelich, Siavash Raigani, Sharon Geerts, Irene Beijert, Fermin M. Fontan, Mohamed M. Aburawi, Sinan Ozer, Peony Banik, Florence Lin, Murat Karabacak, Ehab O.A. Hafiz, Robert J. Porte, Korkut Uygun, James F. Markmann and Heidi Yeh

J. Clin. Med. 2020, 9(1), 269; https://doi.org/10.3390/jcm9010269 - 18 Jan 2020

Cited by 17 | Viewed by 3959

Abstract

Ex situ machine perfusion is a promising technology to help improve organ viability prior to transplantation. However, preclinical studies using discarded human livers to evaluate therapeutic interventions and optimize perfusion conditions are limited by significant graft heterogeneity. In order to improve the efficacy [...] Read more.

Ex situ machine perfusion is a promising technology to help improve organ viability prior to transplantation. However, preclinical studies using discarded human livers to evaluate therapeutic interventions and optimize perfusion conditions are limited by significant graft heterogeneity. In order to improve the efficacy and reproducibility of future studies, a split-liver perfusion model was developed to allow simultaneous perfusion of left and right lobes, allowing one lobe to serve as a control for the other. Eleven discarded livers were surgically split, and both lobes perfused simultaneously on separate perfusion devices for 3 h at subnormothermic temperatures. Lobar perfusion parameters were also compared with whole livers undergoing perfusion. Similar to whole-liver perfusions, each lobe in the split-liver model exhibited a progressive decrease in arterial resistance and lactate levels throughout perfusion, which were not significantly different between right and left lobes. Split liver lobes also demonstrated comparable energy charge ratios. Ex situ split-liver perfusion is a novel experimental model that allows each graft to act as its own control. This model is particularly well suited for preclinical studies by avoiding the need for large numbers of enrolled livers necessary due to the heterogenous nature of discarded human liver research. Full article

(This article belongs to the Special Issue Machine Perfusion and Its Effects on Ischemia-Reperfusion Injury in the Era of Organ Shortage in Solid Organ Transplantation)

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Figure 1
Splitting of discarded donor liver. (A) The whole graft is thoroughly explored, and dissection is performed to the level of the bifurcation of the hepatic artery (HA), portal vein (PV), and bile duct (BD). (B) An anatomical split is performed, with assurance of adequate inflow and outflow. (C) The right lobar and (D) left lobar HA, PV, and BD are cannulated. Full article ">Figure 2
Median vascular resistance of splits lobes during perfusion is statistically the same. (A) Hepatic arterial resistance was qualitatively higher in the right lobes compared with the left lobes, though did not reach statistical significance at any time point. Interestingly, whole-graft arterial resistance was approximately the average of the individual lobes. (B) Split right lobes tended to have higher portal venous resistance during perfusion compared with left lobes (not significant throughout), while whole grafts had lower overall portal venous resistance than individual split lobes. Full article ">Figure 3
Hepatocyte injury and functional recovery are similar between split lobes during perfusion. (A) ALT (alanine aminotransferase) levels were comparable during perfusion between split right and left lobes (no significant difference). Whole grafts had lower ALT levels throughout perfusion compared with individual lobes, indicative of the cellular injury induces during surgical splitting. (B). Lactate levels tended to be higher in split left lobes but were not statistically significant from right lobes. Lactate levels decreased slowly during perfusion in both lobes. Whole grafts had lower lactate levels compared with individual lobes. Full article ">Figure 4
Energy status is reflective of metabolism during subnormothermic perfusion despite splitting. During whole- and split-liver perfusions, (A) ATP:ADP, (B) ATP: AMP, and (C) energy charge ratios demonstrated comparable patterns over time, indicating mitochondrial function and restoration of ATP stores. Full article ">Figure 5
Lobar cellular architecture indicates adequate parenchymal perfusion after splitting. Liver lobules were well preserved during perfusion, with mild sinusoidal congestion and cell swelling, but no increase in ischemic necrosis compared with the preperfusion. No differences were detected between right or left lobes of each individual liver. CV, central vein. * indicates areas of sinusoidal congestion. Full article ">

16 pages, 761 KiB

Open AccessArticle

Serum Cartilage Oligomeric Matrix Protein in Late-Stage Osteoarthritis: Association with Clinical Features, Renal Function, and Cardiovascular Biomarkers

by Jana Riegger, Martin Rehm, Gisela Büchele, Hermann Brenner, Klaus-Peter Günther, Dietrich Rothenbacher and Rolf E. Brenner

J. Clin. Med. 2020, 9(1), 268; https://doi.org/10.3390/jcm9010268 - 18 Jan 2020

Cited by 9 | Viewed by 3731

Abstract

This study aimed to assess associations between serum cartilage oligomeric matrix protein (sCOMP) and phenotypic characteristics in late-stage hip and knee Osteoarthritis (OA) as well as its correlation with further serum markers of possible comorbidities in the Ulm Osteoarthritis Study. Moreover, the prognostic [...] Read more.

This study aimed to assess associations between serum cartilage oligomeric matrix protein (sCOMP) and phenotypic characteristics in late-stage hip and knee Osteoarthritis (OA) as well as its correlation with further serum markers of possible comorbidities in the Ulm Osteoarthritis Study. Moreover, the prognostic relevance of preoperative sCOMP concentrations for short-term functionality and pain outcomes after hip or knee joint replacement was explored. Preoperative serum samples and detailed information about the health status (i.e., WOMAC scores, Hannover Functionality Status (FFbH)) of 754 OA patients undergoing total joint replacement were included. Spearman rank-correlation coefficients and multiple linear regression models were used to evaluate the relationships between sCOMP, other serum markers, and health outcomes. There was a significant positive association between sCOMP and markers of renal (cystatin C, creatinine, and eGFR) and cardiac (e.g., NT-proBNP) impairment. Since renal failure might cause accumulation of sCOMP, additional adjustment with eGFR was performed. Preoperative sCOMP levels in knee OA but not hip OA patients were positively associated with FFbH, WOMAC function sub-scale and total WOMAC scale as well as the post-operative WOMAC stiffness sub-scale six months after surgery. Our data clearly demonstrate an association between sCOMP and renal function as well as other confounding factors, which should be considered in future biomarker studies. Full article

(This article belongs to the Special Issue Orthopaedics: Medicine and Mechanisms)

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12 pages, 767 KiB

Open AccessReview

The Vasoactive Mas Receptor in Essential Hypertension

by Amalie L. Povlsen, Daniela Grimm, Markus Wehland, Manfred Infanger and Marcus Krüger

J. Clin. Med. 2020, 9(1), 267; https://doi.org/10.3390/jcm9010267 - 18 Jan 2020

Cited by 55 | Viewed by 9281

Abstract

The renin–angiotensin–aldosterone system (RAAS) has been studied extensively, and with the inclusion of novel components, it has become evident that the system is much more complex than originally anticipated. According to current knowledge, there are two main axes of the RAAS, which counteract [...] Read more.

The renin–angiotensin–aldosterone system (RAAS) has been studied extensively, and with the inclusion of novel components, it has become evident that the system is much more complex than originally anticipated. According to current knowledge, there are two main axes of the RAAS, which counteract each other in terms of vascular control: The classical vasoconstrictive axis, renin/angiotensin-converting enzyme/angiotensin II/angiotensin II receptor type 1 (AT₁R), and the opposing vasorelaxant axis, angiotensin-converting enzyme 2/angiotensin-(1-7)/Mas receptor (MasR). An abnormal activity within the system constitutes a hallmark in hypertension, which is a global health problem that predisposes cardiovascular and renal morbidities. In particular, essential hypertension predominates in the hypertensive population of more than 1.3 billion humans worldwide, and yet, the pathophysiology behind this multifactorial condition needs clarification. While commonly applied pharmacological strategies target the classical axis of the RAAS, discovery of the vasoprotective effects of the opposing, vasorelaxant axis has presented encouraging experimental evidence for a new potential direction in RAAS-targeted therapy based on the G protein-coupled MasR. In addition, the endogenous MasR agonist angiotensin-(1-7), peptide analogues, and related molecules have become the subject of recent studies within this field. Nevertheless, the clinical potential of MasR remains unclear due to indications of physiological-biased activities of the RAAS and interacting signaling pathways. Full article

(This article belongs to the Special Issue Cardiovascular Disease: From Molecular Mechanisms to Clinical Therapies)

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19 pages, 11621 KiB

Open AccessArticle

Exogenous Gonadotrophin Stimulation Induces Partial Maturation of Human Sertoli Cells in a Testicular Xenotransplantation Model for Fertility Preservation

by Marsida Hutka, Lee B. Smith, Ellen Goossens, W. Hamish B. Wallace, Jan-Bernd Stukenborg and Rod T. Mitchell

J. Clin. Med. 2020, 9(1), 266; https://doi.org/10.3390/jcm9010266 - 18 Jan 2020

Cited by 12 | Viewed by 5082

Abstract

The future fertility of prepubertal boys with cancer may be irreversibly compromised by chemotherapy and/or radiotherapy. Successful spermatogenesis has not been achieved following the xenotransplantation of prepubertal human testis tissue, which is likely due to the failure of somatic cell maturation and function. [...] Read more.

The future fertility of prepubertal boys with cancer may be irreversibly compromised by chemotherapy and/or radiotherapy. Successful spermatogenesis has not been achieved following the xenotransplantation of prepubertal human testis tissue, which is likely due to the failure of somatic cell maturation and function. We used a validated xenograft model to identify the factors required for Leydig and Sertoli cell development and function in immature human testis. Importantly, we compared the maturation status of Sertoli cells in xenografts with that of human testis tissues (n = 9, 1 year-adult). Human fetal testis (n = 6; 14–21 gestational weeks) tissue, which models many aspects of prepubertal testicular development, was transplanted subcutaneously into castrated immunocompromised mice for ~12 months. The mice received exogenous human chorionic gonadotropin (hCG; 20IU, 3×/week). In xenografts exposed continuously to hCG, we demonstrate the maintenance of Leydig cell steroidogenesis, the acquisition of features of Sertoli cell maturation (androgen receptor, lumen development), and the formation of the blood–testis barrier (connexin 43), none of which were present prior to the transplantation or in xenografts in which hCG was withdrawn after 7 months. These studies provide evidence that hCG plays a role in Sertoli cell maturation, which is relevant for future investigations, helping them generate functional gametes from immature testis tissue for clinical application. Full article

(This article belongs to the Special Issue Approach to Male Infertility and Induction of Spermatogenesis)

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11 pages, 240 KiB

Open AccessArticle

Pregnancy Associated Plasma Protein-A as a Cardiovascular Risk Marker in Patients with Stable Coronary Heart Disease During 10 Years Follow-Up—A CLARICOR Trial Sub-Study

by Erik Nilsson, Jens Kastrup, Ahmad Sajadieh, Gorm Boje Jensen, Erik Kjøller, Hans Jørn Kolmos, Jonas Wuopio, Christoph Nowak, Anders Larsson, Janus Christian Jakobsen, Per Winkel, Christian Gluud, Kasper K Iversen, Johan Ärnlöv and Axel C. Carlsson

J. Clin. Med. 2020, 9(1), 265; https://doi.org/10.3390/jcm9010265 - 18 Jan 2020

Cited by 6 | Viewed by 3274

Abstract

Elevated pregnancy-associated plasma protein A (PAPP-A) is associated with mortality in acute coronary syndromes. Few studies have assessed PAPP-A in stable coronary artery disease (CAD) and results are conflicting. We assessed the 10-year prognostic relevance of PAPP-A levels in stable CAD. The CLARICOR [...] Read more.

Elevated pregnancy-associated plasma protein A (PAPP-A) is associated with mortality in acute coronary syndromes. Few studies have assessed PAPP-A in stable coronary artery disease (CAD) and results are conflicting. We assessed the 10-year prognostic relevance of PAPP-A levels in stable CAD. The CLARICOR trial was a randomized controlled clinical trial including outpatients with stable CAD, randomized to clarithromycin versus placebo. The placebo group constituted our discovery cohort (n = 1.996) and the clarithromycin group the replication cohort (n = 1.975). The composite primary outcome was first occurrence of cardiovascular event or death. In the discovery cohort, incidence rates (IR) for the composite outcome were higher in those with elevated PAPP-A (IR 12.72, 95% Confidence Interval (CI) 11.0–14.7 events/100 years) compared to lower PAPP-A (IR 8.78, 8.25–9.34), with comparable results in the replication cohort. Elevated PAPP-A was associated with increased risk of the composite outcome in both cohorts (discovery Hazard Ratio (HR) 1.45, 95% CI 1.24–1.70; replication HR 1.29, 95% CI 1.10–1.52). In models adjusted for established risk factors, these trends were attenuated. Elevated PAPP-A was associated with higher all-cause mortality in both cohorts. We conclude that elevated PAPP-A levels are associated with increased long-term mortality in stable CAD, but do not improve long-term prediction of death or cardiovascular events when added to established predictors. Full article

(This article belongs to the Special Issue Novel Biomarkers for Heart Disease)

17 pages, 1759 KiB

Open AccessArticle

Expression of Taste Receptor 2 Subtypes in Human Testis and Sperm

by Laura Governini, Bianca Semplici, Valentina Pavone, Laura Crifasi, Camilla Marrocco, Vincenzo De Leo, Elisabeth Arlt, Thomas Gudermann, Ingrid Boekhoff, Alice Luddi and Paola Piomboni

J. Clin. Med. 2020, 9(1), 264; https://doi.org/10.3390/jcm9010264 - 18 Jan 2020

Cited by 35 | Viewed by 6286

Abstract

Taste receptors (TASRs) are expressed not only in the oral cavity but also throughout the body, thus suggesting that they may play different roles in organ systems beyond the tongue. Recent studies showed the expression of several TASRs in mammalian testis and sperm, [...] Read more.

Taste receptors (TASRs) are expressed not only in the oral cavity but also throughout the body, thus suggesting that they may play different roles in organ systems beyond the tongue. Recent studies showed the expression of several TASRs in mammalian testis and sperm, indicating an involvement of these receptors in male gametogenesis and fertility. This notion is supported by an impaired reproductive phenotype of mouse carrying targeted deletion of taste receptor genes, as well as by a significant correlation between human semen parameters and specific polymorphisms of taste receptor genes. To better understand the biological and thus clinical significance of these receptors for human reproduction, we analyzed the expression of several members of the TAS2Rs family of bitter receptors in human testis and in ejaculated sperm before and after in vitro selection and capacitation. Our results provide evidence for the expression of TAS2R genes, with TAS2R14 being the most expressed bitter receptor subtype in both testis tissue and sperm cells, respectively. In addition, it was observed that in vitro capacitation significantly affects both the expression and the subcellular localization of these receptors in isolated spermatozoa. Interestingly, α-gustducin and α-transducin, two Gα subunits expressed in taste buds on the tongue, are also expressed in human spermatozoa; moreover, a subcellular redistribution of both G protein α-subunits to different sub-compartments of sperm was registered upon in vitro capacitation. Finally, we shed light on the possible downstream transduction pathway initiated upon taste receptor activation in the male reproductive system. Performing ultrasensitive droplets digital PCR assays to quantify RNA copy numbers of a distinct gene, we found a significant correlation between the expression of TAS2Rs and TRPM5 (r = 0.87), the cation channel involved in bitter but also sweet and umami taste transduction in taste buds on the tongue. Even if further studies are needed to clarify the precise functional role of taste receptors for successful reproduction, the presented findings significantly extend our knowledge of the biological role of TAS2Rs for human male fertility. Full article

(This article belongs to the Special Issue Approach to Male Infertility and Induction of Spermatogenesis)

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18 pages, 3773 KiB

Open AccessArticle

Ringer’s Lactate Prevents Early Organ Failure by Providing Extracellular Calcium

by Biswajit Khatua, Jordan R. Yaron, Bara El-Kurdi, Sergiy Kostenko, Georgios I. Papachristou and Vijay P. Singh

J. Clin. Med. 2020, 9(1), 263; https://doi.org/10.3390/jcm9010263 - 18 Jan 2020

Cited by 34 | Viewed by 8105

Abstract

Objective: Ringer’s lactate may improve early systemic inflammation during critical illnesses like severe acute pancreatitis, which are associated with hypocalcemia. Ringer’s lactate is buffered and contains lactate and calcium. We, thus analyzed extracellular calcium or lactate’s effects on the mechanisms, intermediary markers, and [...] Read more.

Objective: Ringer’s lactate may improve early systemic inflammation during critical illnesses like severe acute pancreatitis, which are associated with hypocalcemia. Ringer’s lactate is buffered and contains lactate and calcium. We, thus analyzed extracellular calcium or lactate’s effects on the mechanisms, intermediary markers, and organ failure in models mimicking human disease with nonesterified fatty acid (NEFA) elevation. Methods: Meta-analyses and experimental studies were performed. Experimentally, extracellular calcium and lactate were compared in their interaction with linoleic acid (LA; a NEFA increased in human severe pancreatitis), and its subsequent effects on mitochondrial depolarization and cytosolic calcium signaling resulting in cell injury. In vivo, the effect of LA was studied on organ failure, along with the effect of calcium or lactate (pH 7.4) on severe acute pancreatitis-associated organ failure. A meta-analysis of human randomized control trials comparing Ringer’s lactate to normal saline was done, focusing on necrosis and organ failure. Results: Calcium reacted ionically with LA and reduced lipotoxic necrosis. In vivo, LA induced organ failure and hypocalcemia. During severe pancreatitis, calcium supplementation in saline pH 7.4, unlike lactate, prevented hypocalcemia, increased NEFA saponification, reduced circulating NEFA and C-reactive protein, reduced pancreatic necrosis adjacent to fat necrosis, and normalized shock (carotid pulse distension) and blood urea nitrogen elevation on day 1. This, however, did not prevent the later increase in serum NEFA which caused delayed organ failure. Meta-analysis showed Ringer’s lactate reduced necrosis, but not organ failure, compared with normal saline. Conclusion: Hypocalcemia occurs due to excess NEFA binding calcium during a critical illness. Ringer’s lactate’s early benefits in systemic inflammation are by the calcium it provides reacting ionically with NEFA. This, however, does not prevent later organ failure from sustained NEFA generation. Future studies comparing calcium supplemented saline resuscitation to Ringer’s lactate may provide insights to this pathophysiology. Full article

(This article belongs to the Section Emergency Medicine)

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(A–C): Isothermal titration calorimetry showing Linoleic acid (LA) reacts with calcium but not lactate. (A) Thermogram showing heat rate of injection of calcium chloride (10 μM/injection) into 1 mM Linoleic acid (LA). Both were dissolved in 10 mM HEPES (pH 7.4) (B) Enthalpograms of injection of sodium lactate (Na-lactate, red line on top) or calcium chloride (CaCl2; green line below) into linoleic acid. The various thermodynamic variables (Mean ± SD (standard deviation)) from three different experiments are mentioned in the table adjacent to the thermogram. (C) Thermogram showing heat rate of injection of sodium lactate (top, red line; 10 μM/injection) and other control experimental conditions below this (black lines). (D–G): Effects of intraperitoneal administration of linoleic acid (LA, 0.1% body weight) to CD-1 mice on their serum calcium (D), carotid pulse distension (pulse dist.) (E), rectal temperature (F), and blood urea nitrogen (Sr. BUN) (G). The vitals were measured at 40 h, and blood parameters at necropsy.* indicates a p-value of <0.05 on Mann-Whitney test. Full article ">Figure 2
Extracellular calcium reduces LA-induced acinar cell injury in vitro. (A) Time course of the effect of different durations of preincubating LA with calcium at 37 °C prior to exposure to primary pancreatic acini, showing its impact on LDH release into the medium of the acini. (B) Extracellular calcium dose-dependently reduces 100 μM LA-induced acinar LDH release over 4 h (red bars) without affecting control cell viability (blue bars). (C) Supplementation with 10 mM sodium lactate had no effect on either baseline cell viability or 100 μM LA-induced acinar cell injury. Effect of different doses of extracellular calcium (0 mM red line, 1 mM green line, 2 mM purple line) on 100 μM LA-induced intracellular calcium elevation (D) and mitochondrial depolarization (E). (F) Chelation of extracellular calcium (1 mM) with EGTA (1 mM) to nominally absent levels increased LA-induced acinar cell injury (dotted red line) at 2 and 4 h. (G): Extracellular calcium dose-dependently reduces 100 μM LA-induced LDH release from HEK293 cells over 4 h (red bars) without affecting control cell viability (blue bars). Full article ">Figure 3
Effect of altering calcium on caerulein-induced (A–D), and changing Cai on LA-induced (E–H) acinar cell injury. (A) Extracellular calcium (1, 2 mM) had no effect on intracellular calcium peak and plateau levels induced by 100 nM caerulein and (B) did not modulate the lack of mitochondrial depolarization with caerulein treatment. (C) Extracellular calcium did not reduce caerulein-induced acinar cell injury, while (D) intracellular calcium chelation with BAPTA-AM (50 μM) or ryanodine receptor antagonization with dantrolene (100 μM) caused caerulein-induced injury to be nonsignificant versus control. (E) BAPTA-AM (50 μM), (F) Dantrolene (100 μM), and (G) thapsigargin (1 μM) all reduced intracellular calcium elevation after 100 μM LA treatment, while BAPTA-AM (E’) partially limiting mitochondrial depolarization Dantrolene (F’) and thapsigargin (G’) had no effect on mitochondrial depolarization. (H): Despite effects on calcium elevation, BAPTA-AM, Dantrolene, and Thapsigargin did not significantly affect LA-induced injury in acinar cells, as measured by LDH release. Full article ">Figure 4
Comparison of the therapeutic calcium and lactate on fat necrosis during severe experimental pancreatitis: (A) serum lipase levels as measured under basal (Bas) conditions before induction of pancreatitis and after 6 h of pancreatitis (6 H), following which either Sodium lactate or Calcium chloride were given intraperitoneally, as described in the methods section. (B) Gross appearance of the peritoneal cavity at necropsy of mice with caerulein pancreatitis alone (CER), caerulein pancreatitis treated with sodium lactate (CER + lactate), or caerulein pancreatitis treated with calcium chloride (CER + calcium). Note the much larger extent of white- colored saponification of fat necrosis in the calcium-treated group (black arrows). (C) Box plots comparing the extent of peri-fat acinar necrosis (% PFAN) measured as a percentage of total pancreatic parenchymal area in the CER + lactate (CER + Lac)- and CER + calclium (CER + Ca)-treated groups. * indicates a p < 0.05. (D) Histologic images of the pancreas and adjacent fat necrosis stained with hematoxylin and eosin (H&E), and von-Kossa for calcium, in the CER + lactate and CER + calcium groups. To the right are zoomed-in images of the inset boxes highlighting the peri-fat acinar necrosis adjacent to the fat necrosis. The scale bar is 200 μm. Fat necrosis resulting in binding of calcium to the NEFA generated is seen as the pinkish stain in adipose tissue on H&E. Note the higher intensity of this in the CER + calcium group using both types of staining, especially von-Kossa, which extends into the parenchyma as a sharply demarcated line (black arrows). The peri-fat acinar necrosis is seen as loss of acinar cell outline and diffuse pale pinkish appearance of acini replacing the normal intensely pink zymogen and blue-colored basal cytoplasm. In the CER + calcium group, which survive longer, the prolonged caerulein stimulation results in acinar ductal metaplasia-like appearance with large central lumens replacing the zymogen (shown as *). Full article ">Figure 5
Effect of no treatment (black), calcium (green), or lactate treatment (red) on parameters of systemic injury and inflammation: (A) Kaplan–Meyer survival curve of the various groups with pancreatitis * indicates a p < 0.05 in the CER + calcium (CER + Ca2+)-treated group vs. other groups. (B) serum calcium and (C) Serum NEFA, as measured at each day of pancreatitis. * indicates a p < 0.05 in the CER + calcium treated groups vs. other groups on ANOVA. # indicates a significant difference from baseline (day 0). (D) Blood C-reactive protein (CRP) after 24 h of pancreatitis. * indicates a significant increase in the group on ANOVA vs. control mice without pancreatitis. (E) Blood urea nitrogen (BUN) and (F) Carotid artery pulse distention (pulse dist.), as measured on each day of pancreatitis. * indicates a p < 0.05 in the cer + calcium-treated groups vs. other groups on ANOVA. Interleukin 6 (IL-6) (G) and tumor necrosis factor (TNF-α) (H) blood levels, one day after pancreatitis initiation (each title followed by 24), and on the post-mortem blood samples. * indicates a significant increase on ANOVA for that time point vs. control. Full article ">Figure 6
Meta-analysis of three randomized controlled trials comparing Ringer’s lactate to normal saline. (A) Study characteristics of those included in the meta-analysis. RL: Ringer’s lactate. NS: Normal Saline. AP: Acute pancreatitis. AVG: Average. SD: Standard deviation. NS: Not significant. (B) Outcomes of acute pancreatitis after hydration with Ringer’s lactate vs. normal saline. AP: Acute pancreatitis. RL: Ringer’s lactate NS: Normal Saline. Full article ">Figure 6 Cont.
Meta-analysis of three randomized controlled trials comparing Ringer’s lactate to normal saline. (A) Study characteristics of those included in the meta-analysis. RL: Ringer’s lactate. NS: Normal Saline. AP: Acute pancreatitis. AVG: Average. SD: Standard deviation. NS: Not significant. (B) Outcomes of acute pancreatitis after hydration with Ringer’s lactate vs. normal saline. AP: Acute pancreatitis. RL: Ringer’s lactate NS: Normal Saline. Full article ">

16 pages, 4140 KiB

Open AccessArticle

Effect of Le Fort I Maxillary Advancement and Clockwise Rotation on the Anteromedial Cheek Soft Tissue Change in Patients with Skeletal Class III Pattern and Midface Deficiency: A 3D Imaging-Based Prediction Study

by Hsin-Chih Lai, Rafael Denadai, Cheng-Ting Ho, Hsiu-Hsia Lin and Lun-Jou Lo

J. Clin. Med. 2020, 9(1), 262; https://doi.org/10.3390/jcm9010262 - 18 Jan 2020

Cited by 19 | Viewed by 8247

Abstract

Patients with a skeletal Class III deformity may present with a concave contour of the anteromedial cheek region. Le Fort I maxillary advancement and rotational movements correct the problem but information on the impact on the anteromedial cheek soft tissue change has been [...] Read more.

Patients with a skeletal Class III deformity may present with a concave contour of the anteromedial cheek region. Le Fort I maxillary advancement and rotational movements correct the problem but information on the impact on the anteromedial cheek soft tissue change has been insufficient to date. This three-dimensional (3D) imaging-assisted study assessed the effect of surgical maxillary advancement and clockwise rotational movements on the anteromedial cheek soft tissue change. Two-week preoperative and 6-month postoperative cone-beam computed tomography scans were obtained from 48 consecutive patients who received 3D-guided two-jaw orthognathic surgery for the correction of Class III malocclusion associated with a midface deficiency and concave facial profile. Postoperative 3D facial bone and soft tissue models were superimposed on the corresponding preoperative models. The region of interest at the anteromedial cheek area was defined. The 3D cheek volumetric change (mm³; postoperative minus preoperative models) and the preoperative surface area (mm²) were computed to estimate the average sagittal movement (mm). The 3D cheek mass position from orthognathic surgery-treated patients was compared with published 3D normative data. Surgical maxillary advancement (all p < 0.001) and maxillary rotation (all p < 0.006) had a significant effect on the 3D anteromedial cheek soft tissue change. In total, 78.9%, 78.8%, and 78.8% of the variation in the cheek soft tissue sagittal movement was explained by the variation in the maxillary advancement and rotation movements for the right, left, and total cheek regions, respectively. The multiple linear regression models defined ratio values (relationship) between the 3D cheek soft tissue sagittal movement and maxillary bone advancement and rotational movements of 0.627 and 0.070, respectively. Maxillary advancements of 3–4 mm and >4 mm resulted in a 3D cheek mass position (1.91 ± 0.53 mm and 2.36 ± 0.72 mm, respectively) similar (all p > 0.05) to the 3D norm value (2.15 ± 1.2 mm). This study showed that both Le Fort I maxillary advancement and rotational movements affect the anteromedial cheek soft tissue change, with the maxillary advancement movement presenting a larger effect on the cheek soft tissue movement than the maxillary rotational movement. These findings can be applied in future multidisciplinary-based decision-making processes for planning and executing orthognathic surgery. Full article

(This article belongs to the Section Dentistry, Oral Surgery and Oral Medicine)

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47 pages, 1510 KiB

Open AccessReview

Glial Cells—The Strategic Targets in Amyotrophic Lateral Sclerosis Treatment

by Tereza Filipi, Zuzana Hermanova, Jana Tureckova, Ondrej Vanatko and Miroslava Anderova

J. Clin. Med. 2020, 9(1), 261; https://doi.org/10.3390/jcm9010261 - 18 Jan 2020

Cited by 52 | Viewed by 9650

Abstract

Amyotrophic lateral sclerosis (ALS) is a fatal neurological disease, which is characterized by the degeneration of motor neurons in the motor cortex and the spinal cord and subsequently by muscle atrophy. To date, numerous gene mutations have been linked to both sporadic and [...] Read more.

Amyotrophic lateral sclerosis (ALS) is a fatal neurological disease, which is characterized by the degeneration of motor neurons in the motor cortex and the spinal cord and subsequently by muscle atrophy. To date, numerous gene mutations have been linked to both sporadic and familial ALS, but the effort of many experimental groups to develop a suitable therapy has not, as of yet, proven successful. The original focus was on the degenerating motor neurons, when researchers tried to understand the pathological mechanisms that cause their slow death. However, it was soon discovered that ALS is a complicated and diverse pathology, where not only neurons, but also other cell types, play a crucial role via the so-called non-cell autonomous effect, which strongly deteriorates neuronal conditions. Subsequently, variable glia-based in vitro and in vivo models of ALS were established and used for brand-new experimental and clinical approaches. Such a shift towards glia soon bore its fruit in the form of several clinical studies, which more or less successfully tried to ward the unfavourable prognosis of ALS progression off. In this review, we aimed to summarize current knowledge regarding the involvement of each glial cell type in the progression of ALS, currently available treatments, and to provide an overview of diverse clinical trials covering pharmacological approaches, gene, and cell therapies. Full article

(This article belongs to the Special Issue Glial Cells in Central Nervous System (CNS) Pathology and Repair)

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18 pages, 507 KiB

Open AccessArticle

Clinical Factors Associated with Adherence to the Follow-Up Examination after Positive Fecal Occult Blood Test in National Colorectal Cancer Screening

by Byung Chang Kim, Minjoo Kang, Eunjung Park, Jeong-Im Shim, Shinhee Kang, Jessie Lee, Ha Jin Tchoe, Kyeong Ae Kong, Duk Hwan Kim, Yu Jin Kim, Kui Son Choi and Chang Mo Moon

J. Clin. Med. 2020, 9(1), 260; https://doi.org/10.3390/jcm9010260 - 18 Jan 2020

Cited by 11 | Viewed by 3253

Abstract

Background: The compliance with the follow-up examination after a positive fecal occult blood test (FOBT) is lower than expected. We aimed to evaluate the adherence rate to the follow-up examination in patients with a positive FOBT and to identify the clinical factors associated [...] Read more.

Background: The compliance with the follow-up examination after a positive fecal occult blood test (FOBT) is lower than expected. We aimed to evaluate the adherence rate to the follow-up examination in patients with a positive FOBT and to identify the clinical factors associated with this adherence. Methods: The study population comprised adults aged ≥50 years who participated in the National Cancer Screening Program for colorectal cancer (CRC) in 2013. Compliance was defined as undergoing follow-up examination within 1 year of a positive FOBT. Results: From 214,131 individuals with a positive FOBT, 120,911 (56.5%) were in the compliance group and 93,220 (43.5%) were in the non-compliance group. On multivariate analysis, good compliance was associated with men (odds ratio (OR) = 1.12, 95% confidence interval (CI) (1.09–1.15)), younger ages (70–79 years, OR = 2.19 (2.09–2.31); 60–69 years, OR = 3.29 (3.13–3.46); 50–59 years, OR = 3.57 (3.39–3.75) vs. >80 years), previous experience of CRC screening (a negative FOBT, OR = 1.18 (1.15–1.21); a positive FOBT, OR = 2.42 (2.31–2.54)), absent previous experience of colonoscopy or barium enema (OR = 2.06 (1.99–2.13)), higher economic income (quartile, 75%, OR = 1.14 (1.11–1.17); 100%, OR = 1.22 (1.19–1.25)), current smokers (OR = 1.12 (1.09–1.15)), alcohol intake (OR = 1.03 (1.01–1.05)), active physical activity (≥3 times/week, OR = 1.13 (1.11–1.15)), depression (OR = 1.11 (1.08–1.14)), and present comorbidities (Charlson Comorbidity Index, ≥1). Conclusion: This study identified clinical factors, namely, male, younger ages, prior experience of fecal test, absent history of colonoscopy or double-contrast barium enema (DCBE) within 5 years, and high socioeconomic status to be associated with good adherence to the follow-up examination after a positive FOBT. Full article

(This article belongs to the Section Oncology)

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14 pages, 1228 KiB

Open AccessArticle

Ipragliflozin Additively Ameliorates Non-Alcoholic Fatty Liver Disease in Patients with Type 2 Diabetes Controlled with Metformin and Pioglitazone: A 24-Week Randomized Controlled Trial

by Eugene Han, Yong-ho Lee, Byung-Wan Lee, Eun Seok Kang and Bong-Soo Cha

J. Clin. Med. 2020, 9(1), 259; https://doi.org/10.3390/jcm9010259 - 18 Jan 2020

Cited by 52 | Viewed by 4595

Abstract

Despite the benefits of pioglitazone in the treatment of non-alcoholic fatty liver disease (NAFLD), many treated patients continue to experience disease progression. We aimed to investigate the additive effect of ipragliflozin on NAFLD in patients with type 2 diabetes treated with metformin and [...] Read more.

Despite the benefits of pioglitazone in the treatment of non-alcoholic fatty liver disease (NAFLD), many treated patients continue to experience disease progression. We aimed to investigate the additive effect of ipragliflozin on NAFLD in patients with type 2 diabetes treated with metformin and pioglitazone. In this 24-week randomized controlled trial, 44 patients with type 2 diabetes and comorbid NAFLD were either randomized to receive 50 mg/day of ipragliflozin as an add-on treatment (n = 29) or maintained on metformin and pioglitazone (n = 15). The fatty burden was assessed using the fatty liver index, NAFLD liver fat score, and controlled attenuation parameter (CAP). Changes in fat and muscle depots were measured by dual-energy x-ray absorptiometry and abdominal computed tomography scans. The enrolled patients were relatively controlled (mean baseline glycated hemoglobin of 6.6% ± 0.6%) and centrally obese (mean waist circumference of 101.6 ± 10.9 cm). At week 24, patients in the ipragliflozin add-on group exhibited reduced hepatic fat content (fatty liver index: −9.8 ± 1.9, p = 0.002; NAFLD liver fat score: −0.5 ± 0.2, p = 0.049; CAP: −8.2 ± 7.8 dB/m², p = 0.133). Ipragliflozin add-on therapy also reduced whole-body visceral fat and the ratio of visceral to subcutaneous fat (change in whole-body visceral fat: −69.6 ± 21.5 g; change in abdominal visceral fat: −26.2 ± 3.7 cm²; abdominal visceral to subcutaneous fat ratio: −0.15 ± 0.04; all p < 0.05). In conclusion, ipragliflozin treatment significantly ameliorates liver steatosis and reduces excessive fat in euglycemic patients with type 2 diabetes and NAFLD taking metformin and pioglitazone. Full article

(This article belongs to the Special Issue Pathophysiology and Complications of Type 2 Diabetes Mellitus)

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Changes in body weight, body mass index, waist circumference, abdominal adipose tissue, body fat and muscle from baseline to week 24. (A) Body weight. (B) Body mass index. (C) Waist circumference. (D) Abdominal visceral adipose tissue assessed by CT. (E) Abdominal subcutaneous adipose tissue assessed by CT. (F) Abdominal visceral to subcutaneous adipose tissue ratio assessed by CT. Red square: metformin + pioglitazone + ipragliflozin. Black circle: metformin + pioglitazone. * p < 0.05. Full article ">Figure 2
Changes in body fat, muscle and hepatic fat contents from baseline to week 24. (A) Total body fat. (B) Visceral body fat. (C) Total muscle mass by dual-energy x-ray absorptiometry. (D) Controlled attenuation parameter assessed by transient liver elastography. (E) Fatty liver index. (F) Non-alcoholic fatty liver disease liver fat score. Red square: metformin + pioglitazone + ipragliflozin. Black circle: metformin + pioglitazone. * p < 0.05. Full article ">Figure 3
Correlation between changes in body weight, adipose tissue, liver fat and muscle mass. (A) Correlation between changes in visceral adipose tissue and body weight. (B) Correlation between changes in controlled attenuation parameter and body weight. (C) Correlation between changes in visceral adipose tissue and muscle mass. (D) Correlation between changes in visceral adipose tissue and subcutaneous adipose tissue. Full article ">

13 pages, 1161 KiB

Open AccessArticle

The Effect of Proton Pump Inhibitor Use on Renal Function in Kidney Transplanted Patients

by Dominik J. G. Flothow, Barbara Suwelack, Hermann Pavenstädt, Katharina Schütte-Nütgen and Stefan Reuter

J. Clin. Med. 2020, 9(1), 258; https://doi.org/10.3390/jcm9010258 - 18 Jan 2020

Cited by 8 | Viewed by 3888

Abstract

Recently, proton pump inhibitor (PPI) intake has been linked to acute kidney injury and chronic kidney disease. The objective of this study was to assess the effect of PPIs on renal function and rejection rate in kidney transplant patients. We performed a single [...] Read more.

Recently, proton pump inhibitor (PPI) intake has been linked to acute kidney injury and chronic kidney disease. The objective of this study was to assess the effect of PPIs on renal function and rejection rate in kidney transplant patients. We performed a single center, retrospective analysis of 455 patients who received a kidney transplant between May 2010 and July 2015. Median follow-up time was 3.3 years. PPI prescription was assessed in half-year intervals. Primary outcome parameters were the estimated glomerular filtration rate (eGFR), change in the eGFR, and >30% and >50% eGFR decline for different time periods (up to four years post-transplantation). Our secondary outcome parameter was occurrence of biopsy proven acute rejection (BPAR) in the first two years after transplantation. Except for >30% eGFR decline from half a year to two years post-transplantation (p = 0.044) and change in the eGFR, >30% and >50% eGFR decline showed no association with PPI intake in our patient cohort (p > 0.05). Similarly, by analyzing 158 rejection episodes, BPAR showed no correspondence with mean daily PPI intake. We conclude that prolonged PPI intake has no relevant adverse effect on kidney transplant function or rejection rates. Polypharmacy, however, remains a problem in renal transplant recipients and it is thus advisable to question the necessity of PPI prescriptions when clear indications are missing. Full article

(This article belongs to the Special Issue Recent Advances and Clinical Outcomes of Kidney Transplantation)

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Trend in mean estimated glomerular filtration rate (eGFR). Mean eGFR is plotted against time in the two patient groups. Grouping is according to PPI intake or non-intake at half a year post-transplantation. Full article ">Figure 2
Results of the biopsy proven rejection analyses are shown. Grouping is according to PPI intake (pantoprazole equivalent) in the relevant time period. Rejection prevalence is shown as absolute number and as percentage of patients in the group. The dark part of each bar represents the number of patients with rejections. Abbreviations: n, total number of patients in the group; Rej, number of patients who experienced a graft rejection; pTx, post transplantation. (A) Analysis of rejections in months 1–6 pTx. (B) Analysis of rejections in months 7–12 pTx. (C) Analysis of rejections months 13–24 pTx. Full article ">Figure 2 Cont.
Results of the biopsy proven rejection analyses are shown. Grouping is according to PPI intake (pantoprazole equivalent) in the relevant time period. Rejection prevalence is shown as absolute number and as percentage of patients in the group. The dark part of each bar represents the number of patients with rejections. Abbreviations: n, total number of patients in the group; Rej, number of patients who experienced a graft rejection; pTx, post transplantation. (A) Analysis of rejections in months 1–6 pTx. (B) Analysis of rejections in months 7–12 pTx. (C) Analysis of rejections months 13–24 pTx. Full article ">

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J. Clin. Med., Volume 9, Issue 1 (January 2020) – 287 articles

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