Clinical Pathology

BACKGROUND: Hospital-acquired (nosocomial) infection is a common serious health problem worldwide, especially in pediatric intensive care units and is associated with high mortality and morbidity, prolonged hospital stays and high cost.AIM: To determine the types of organisms involved in hospital-acquired an infection in two pediatric intensive care units during the one-year study and its anti-microbial susceptibility.MATERIAL AND METHODS: This study was carried out in the pediatric intensive care units (PICU) of Ain Shams & Cairo Universities, where 86 pediatric patients were recruited. Their age ranged from 1 month to 156 months with mean 20.7 ± 25.8 months. Male to female ratio was 37:29. Four samples were collected from each child for culture and sensitivity: blood, endotracheal aspirate, urine and skin swab.RESULTS: The most common microorganism was staphylococcus while Gram-negative bacteria were the commonest group. Amikacin and imipenem are the most sensitive antibiotics. Ri...

Immunoregulatory cytokines may influence the hepatitis C virus (HCV) infection outcome. This study aimed to determine the genotypic
and allelic frequencies of the interleukin (IL)-10 (-1082) G/A polymorphism, and its association with chronicity or resolution of HCV genotype 4 infection
in Egypt. The frequencies of different dimorphic polymorphisms based on single nucleotide substitution in chronic HCV patients (50) and resolved HCV
patients (50) were: IL-10 (-1082) G/G 22 (44%) and 18 (36%), G/A 19 (38%) and 24 (48%), and A/A 9 (18%), and 8 (16%), respectively. In the sustained
virologic response (SVR) (36) and spontaneously resolved subjects (14) groups, the frequencies were: IL-10 (-1082) G/G 11 (30.6%) and 7 (50%) G/A 18
(50%) and 6 (42.9%), A/A 7 (19.4%) and 1 (7.1%), respectively. An association between male gender and chronic hepatitis C outcome (P value 0.041) was
found. However, no significant gender difference was found when we compared females versus males with elevated alanine transaminase (ALT) levels in
the chronic HCV patient group (P value = 1).
CONCLUSION: No significant difference in the frequency of IL-10 single nucleotide polymorphism (SNP) at position 1082 was found between chronic
and resolved HCV subjects.
KEYWORDS: interleukin-10, single nucleotide polymorphism, chronic hepatitis C, resolved hepatitis C, HCV genotype 4

Introduction: The killer cell immunoglobulin-like receptors (KIR) control the activation or
inhibition of Natural killer (NK) cells by recognition of specific HLA class I ligands, such that target
cell cytolysis occur only when activating signals overcome inhibitory signals. The aim of the study is
to demonstrate the effect of HLA class I and KIR gene interaction on the outcome of HCV infection in
Egypt. Material and methods: The study included 137 individuals with resolved HCV infection; 17
cases spontaneous resolution and 120 cases with sustained virologic response (SVR) and 125
individuals with chronic HCV infection. Individuals with resolved and others with chronic HCV
infection are subjected to genotyping for (HLA class I), (KIR) gene of killer (NK) cells. Results
:(KIR2DL3-HLA-C1) was higher in the group with resolved infection (19.7%) relative to that with
persistent infection (7.2%) P-value=0.003, whereas the inhibitory receptor ligand (KIR2DL1-HLAC2)
was higher in the group with persistent infection (19.2%) compared to that with resolved infection
(9.4%) P-value =0.02. No significant difference was found between the two groups with the following
inhibitory KIR-HLA combinations : KIR2DL2-HLA-C1, KIR2DL2/KIR2DL3-HLA-C1 , KIR2DL3-
HLA-C1/C2. The activating receptor ligand KIR2DS2-HLA-C1, KIR2DS1-HLA-C2 and KIR3DS1-
HLA-Bw4 were not associated with HCV resolution. Conclusion: HLA/KIR genotype plays a role in
determining the outcome of HCV infection. In our population, viral resolution was associated with
KIR2DL3/HLA-C1 homozygosity, while virus persistence was associated with KIR2DL1/HLAC2/
C2.
Key words: HCV, HLA class I, KIR genes

Methicillin-resistant Staphylococci; both methicillin resistant S. aureus (MRSA) and
methicillin resistant coagulase negative Staphylococci (MRCNS) have a worldwide distribution and are important
causes of clinical and epidemiological problems. The aim of this study was to evaluate the usefulness of some
phenotypic methods for detection of methicillin resistant Staphylococci in clinical laboratories; cefoxitin disc
diffusion (CDD), oxacillin resistance screening agar base (ORSAB) and oxacillin E test, in reference to mecA gene
based real-time PCR. Also to study the antibiotic resistance pattern of the methicillin resistant Staphylococci
isolates. Materials and methods: A total of 95 clinical isolates of Staphylococci were tested for methicillin
resistance by CDD test, ORSAB and oxacillin E-test and were compared to mecA based real time PCR as reference
method. Results: MecA gene was detected by PCR in 48/95 (51%) of all Staphylococci isolates; 28/57 (49%) in S.
aureus and 20/38 (53%) in CNS isolates. CCD test showed 100% sensitivity and 98% specificity for detection of
MRSA and MRCNS. ORSAB and Oxacillin E test had 94% and 90% sensitivity and 96% and 98% specificity
respectively. In our study, MRSA isolates were resistant to ciprofloxacin-78%, erythromycin-74%, clindamycin-
71%, Gentamicin-70%, amikacin- 64%, azithromycin-63%, doxycyclin-60%, levofloxacin and linezolid-48%,
trimethoprim-sulfamethoxazole-29% and vancomycin-0%. Conclusion: Cefoxitin disc diffusion can be used as a
reliable conventional, simple and cheap alternative to PCR for detection of MRSA and MRCNS in minimal resources
circumstances. It can also detect other mechanisms of resistance other than mecA. Additional confirmatory test is
needed with oxacillin screening agar and oxacillin E-test for detection of hetero-resistant strains to methicillin.

This study aimed to establish an accurate and sensitive polymerase chain reaction (PCR) technique for the diagnosis of active human brucellosis in Egypt. We failed to extract Brucella DNA with a commercial kit, but an extraction kit designed in-house using 2 sets of primers [B4/B5 (223 bp) and JPF/JPR (193 bp)] was successful and more economical. The technique showed high sensitivity, specificity and accuracy. The PCR positivity increased significantly with increasing seropositivity titres by the standard tube agglutination test and showed 100% positivity in patients with positive blood cultures. We recommend using PCR as an alternative to culture for diagnosis of brucellosis.

Chronic obstructive pulmonary disease (COPD) is a leading cause
of disability and death. The most common cause of COPD is smoking. There is
evidence suggesting that genetic factors influence COPD susceptibility and variants
in several candidate genes have been significantly associated with COPD.
In this study, we aimed to investigate the possible association of the TNF-α
–308, SPB+1580, IL-13 –1055 gene polymorphisms and latent adenovirus C infection
with COPD in an Egyptian population.
Material and methods: Our study included 115 subjects (75 smokers with COPD,
25 resistant smokers and 15 non-smokers) who were subjected to spirometric
measurements, identification of adenovirus C and genotyping of TNF-α –308G/A,
SP-B+1580 C/T and IL-13 –1055 C/T polymorphisms by real-time PCR.
Results: The adenovirus C gene was identified in all subjects. The distribution
of TNF-α genotypes showed no significant differences between different groups.
However, homozygous A genotype was associated with a significant decrease
in FEV1, FEV1/FVC and FEF25/75% of predicted in COPD (p < 0.05). As regards
SP-B genotypes, resistant smokers had a significantly higher homozygous T
genotype frequency compared to COPD and non smokers (p = 0.005). Interleukin
13 genotypes showed no significant difference between different groups.
There was a significant decrease in FEF25/75% of predicted in T allele carriers
in COPD patients (p = 0.001).
Conclusions: The COPD is a disease caused by the interaction of combined genes
and environmental influences, in the presence of smoking and latent adenovirus
C infection, TNF-α –308A, SPB +1580 T and IL-13 –1055 T polymorphisms predispose
to the development of COPD.

Ｏｂｊｅｃｔｉｖｅ：Ｄｉａｇｎｏｓｉｓｏｆａｔｙｐｉｃａｌｐａｔｈｏｇｅｎｓａｓａｎａｅｔｉｏｌｏｇｙｆｏｒｃｏｍｍｕｎｉｔｙａｃｑｕｉｒｅｄｐｎｅｕｍｏｎｉａ（ＣＡＰ）ｉｎｃｈｉｌ
ｄｒｅｎｉｓａｃｈａｌｌｅｎｇｅｗｏｒｌｄｗｉｄｅ．Ｔｈｅａｉｍｏｆｔｈｉｓｓｔｕｄｙｗａｓｔｏｄｅｔｅｃｔｔｈｅｆｒｅｑｕｅｎｃｙｏｆａｔｙｐｉｃａｌｐａｔｈｏｇｅｎｓａｓａ
ｃａｕｓｅｏｆｃｏｍｍｕｎｉｔｙａｃｑｕｉｒｅｄｐｎｅｕｍｏｎｉａ（ＣＡＰ）ｉｎＥｇｙｐｔｉａｎｃｈｉｌｄｒｅｎ．Ｍｅｔｈｏｄｓ：Ｆｒｏｍ５０ｃｈｉｌｄｒｅｎ（ｗｉｔｈａｇｅ
ｒａｎｇｅｄｆｒｏｍ２ｍｏｎｔｈｓｔｏ１２ｙｅａｒｓ）ｈｏｓｐｉｔａｌｉｚｅｄｆｏｒｃｏｍｍｕｎｉｔｙａｃｑｕｉｒｅｄｐｎｅｕｍｏｎｉａ；ｒｅｓｐｉｒａｔｏｒｙｓｐｕｔｕｍｓａｍｐｌｅｓ
ｗｅｒｅｃｏｌｌｅｃｔｅｄｂｙｉｎｄｕｃｔｉｏｎｏｒｓｐｏｎｔａｎｅｏｕｓｌｙ．ＡｌｌｓａｍｐｌｅｓｗｅｒｅｓｕｂｊｅｃｔｅｄｔｏｃｏｎｖｅｎｔｉｏｎａｌｃｕｌｔｕｒｅｓａｎｄＰｏｌｙｍｅｒ
ａｓｅＣｈａｉｎＲｅａｃｔｉｏｎ（ＰＣＲ）ｔｅｃｈｎｉｑｕｅＤＮＡｅｘｔｒａｃｔｉｏｎｆｏｒｉｄｅｎｔｉｆｉｃａｔｉｏｎｏｆＭｙｃｏｐｌａｓｍａ，Ｃｈｌａｍｙｄｉａｐｎｅｕｍｏｎｉａｅ
ａｎｄＬｅｇｉｏｎｅｌｌａｐｎｅｕｍｏｐｈｉｌａ．Ｒｅｓｕｌｔｓ：Ａｄｅｆｉｎｉｔｅｐａｔｈｏｇｅｎｗａｓｉｄｅｎｔｉｆｉｅｄｉｎ７８％ ｏｆｔｈｅｓｔｕｄｉｅｄｃｈｉｌｄｒｅｎ；３０％
ｔｙｐｉｃａｌｂａｃｔｅｒｉａ，８％ ｃａｎｄｉｄａａｌｂｉｃａｎｓａｎｄａｔｙｐｉｃａｌｂａｃｔｅｒｉａｉｎ４０％ ｏｆｔｈｅｐｎｅｕｍｏｎｉｃｃｈｉｌｄｒｅｎ．Ｃｈｌａｍｙｄｉａ
ｐｎｅｕｍｏｎｉａｅｗａｓｉｓｏｌａｔｅｄｆｒｏｍ２６％ ｏｆｔｈｅｃｈｉｌｄｒｅｎｗｈｉｌｅＭｙｃｏｐｌａｓｍａｐｎｅｕｍｏｎｉａｅｗａｓｉｓｏｌａｔｅｄｆｒｏｍ１４％， ｗｈｅｒｅａｓＬｅｇｉｏｎｅｌｌａｐｎｅｕｍｏｐｈｉｌｌａｗａｓｎｏｔｉｓｏｌａｔｅｄａｔａｌｌ．Ｃｏｎｃｌｕｓｉｏｎ：Ａｔｙｐｉｃａｌｐａｔｈｏｇｅｎｓａｒｅｅｖｉｄｅｎｔａｓａｐｏ
ｔｅｎｔｉａｌａｅｔｉｏｌｏｇｙｆｏｒｃｏｍｍｕｎｉｔｙａｃｑｕｉｒｅｄｐｎｅｕｍｏｎｉａｉｎ（１３．３％）ｏｆｙｏｕｎｇａｎｄ（８０％）ｏｆｏｌｄｅｒＥｇｙｐｔｉａｎｃｈｉｌ
ｄｒｅｎ．
Ｋｅｙｗｏｒｄｓ：Ａｔｙｐｉｃａｌｐａｔｈｏｇｅｎｓ；Ｃｏｍｍｕｎｉｔｙａｃｑｕｉｒｅｄｐｎｅｕｍｏｎｉａ；Ｃｈｉｌｄｒｅｎ；Ｅｇｙｐ

Aggressive peridontal disease
characterized by loss of bone around special teeth in
adolescents and young adults.
Objective: The objective of this study was to assess the
clinical and antimicrobial effect of ozonized water in
management of aggressive periodontitis.
Subjects and Methods: Twenty patients suffering from
localized and generalized aggressive periodontitis were
selected, age range from 13 to 25 years. They were clinically
and radiographically examined. Scaling, root planning, oral
hygiene measures with or without ozone application were
followed.
Results: Different treatment plans were evaluated
regarding plaque and gingival indices, pocket depth,
attachment loss and mobility scores. Antibacterial effect of
ozone was investigated.
Conclusion and Recommendation: We recommend the use
of oral hygiene instructions, scaling and root planning together
with subgingival irrigation with ozonized water in the
management of aggressive periodontitis. Up to our knowledge
this is the first Egyptian study using subgingival irrigation in
combination with the routine treatment plans for Aggressive
periodontitis.
Keywords: Aggressive periodontitis - Ozone

We examined the etiology of nosocomial infection in a neonatal intensive care unit and determined
antibiotic susceptibility of the isolated organisms to detect changes in infection patterns. The study included
56 neonates who developed nosocomial infection (10.93±11.2 days) from neonatal intensive care unit, Cairo
University hospital. Four samples were collected from each neonate before treatment: blood, tracheal
aspirate, urine and skin swab. Blood agar, Mac Conkey, CLED agar media and Thioglycolate broth bottles were
used. For rapid identification of enterobactericae family we used API for enteric and non- enteric nonfermenter
organism identification. Antimicrobial susceptibility tests were done. Nosocomial infections
presented as neonatal septicemia (46.4%), pneumonia (14.3%), skin infection (10.7%) and mixed infections
(28.6%). Gram-negative organisms were isolated from 42 neonates (75%), while 20 neonates showed Grampositive
septicemia (35.7%). Major isolated pathogens were Klebsiella species (39.3%), staphylococcus aureus
(21.4%), coagulase negative staphylococci (14.3%), Acinetobacter species (14.3%), and pseudomonas
aeroginosa (7.14%). The most encountered risk factors associated with nosocomial sepsis was prematurity
(adjusted OR=9.444, 95CI=1.433-62.238). The Gram negative and positive organisms showed a high degree of
resistance to commonly used antibiotics: third generation cephalosporines, amoxicillin/clavulanic acid,
oxacillin, ciprofloxacin, gentamycin and trimethoprim/sulphamethaxzole. Both types of organisms were
sensitive to amikacin and imipenem.
Keywords: neonates, nosocomial infection, bacteria, antibiotic resistance

An imbalance between the Th1 and Th2 cytokines indicates the
development of the fibrotic response. This study focus on the mediators that
influence the imbalance in pulmonary fibrosis by assessing the variation in the
serum level of IL-2 (type-1 cytokine) and IL-8 (type-2 cytokine).
Material and methods: This study included 25 patients with lung fibrosis and ten
healthy children as a control group. The lung fibrosis group was subdivided into diffuse
and localized according to the pattern of fibrosis. Causes of diffuse lung fibrosis were
SLE or post chemotherapy course due to malignancies; those of localized fibrosis
were unresolved pneumonia or old pulmonary tuberculosis. There was a significant
increase in serum level of IL-8 and an insignificant increase in serum level of IL-2 in
pulmonary fibrosis group compared with the control (p=0.02, p=0.06) respectively.
Results: There was a significant increase in the serum level of IL-2 in diffuse lung
fibrosis group compared with that of control (p=0.004) and localized lung fibrosis
(p=0.01) groups. There was no increase in the serum level of IL-8 in diffuse lung
fibrosis group compared with that of control group (p=0.9). The localized lung
fibrosis group shown significant increase in the serum level of IL-8 compared with
the control (p=0.002) and the diffuse lung fibrosis groups (p=0.02) respectively.
Conclusions: In conclusion, the imbalance between IL-8 (Th2) and IL-2 (Th1)
cytokines in pulmonary fibrosis with the predominance of Th2, as compared to Th1
cytokines is important in progression of fibrosis.
Key words: cytokines, interleukin, pulmonary fibrosis, T-helper

CRP is a commonly used marker of inflam-mation to select serious bacterial infections (SBI). Difficult accessibility in segregated districts clinics lowers of its value for rapid prediction and start of antibiotics. A bed site rapid test is supposed to solve this problem provided having at least the same accuracy in documenting bacterial infection.
Patients and Methods: 96 patients of pediatric age group presenting with acute infections were subjected to CRP laboratory and radioimmunoassay Actim CRP rapid test. The definitive diagnosis was confirmed without both tests inter-ference.
Results: Of the 96 cases, 49 had simple infections and 47 SBI. Of them, 26 patients had streptococcal pharyngo-tonsillitis, 10 patients had bacterial pneumonia and 11 patients had sepsis. The sensitivity, specificity, negative predictive value, positive predictive value of CRP to serious bacterial infection were 91.5%, 59.2%, 68.3% and 87.9% respectively. While that of Actim rapid test were 89.4%, 63.3%, 70.0% and 86.1% respectively. The receiver-operating characteristic (ROC) area under the curve for CRP laboratory was 75.3% and that for Actim 76.3%.
Conclusion: CRP laboratory and Actim are quite similar in predicting SBI. However, Actim is slightly more accurate and definitively more practical as regard time and space. Serial evaluation of rapid test increases of its sensitivity and negative predictive value to SBI. We can accept a low speci-ficity and positive predictive value as the test is affected by non serious bacterial infection.

Acute streptococcal pharyngitis represents a high prevalence in developing countries responsible of morbidity and mortality. Difficulty of prompt diagnosis is responsible for either inadequate therapy and subsequent rheumatic heart disease and renal failure, or abuse of antibi-otics. Group AB hemolytic streptococcus (GABHS) detection by rapid tests is useful due to its rapidity and relative low cost. However sensitivity and specificity remain debatable. In Egypt, suspicion about its accuracy remains an important obstacle for mass utilization. The objective of this study was to evaluate the accuracy of a GABHS rapid antigen detection test kit, in comparison with oropharynx swab culture.
Methods: Children aged 3 to 15 years with clinical diag-noses of non treated acute pharyngitis were recruited at public emergency health centers in a rural region eastern Cairo, with 95 patients, subjected to oropharyngeal swabs for rapid GABHS testing and conventional culture.
Results: We observed sensitivity of 80.65%, specificity of 96.88%, a positive predictive value of 92.59%, a negative predictive value of 91.18% and a positive likelihood ratio for the rapid test used here, compared with throat culture. It was found a group of any 3 of fever, sore throat, dysphagia, cervical lymphadenopathy always present in false negative cases.
Conclusions: The rapid test studied exhibited a good correlation with culture and is, therefore, of great use in clinical practice for detection of GABHS. Clinical picture has to be taken in consideration to increase the sensitivity of the test.
Key Words: Streptococcal – Pharyngo-tonsillitis

Oxidative stress may play a pathogenic role in chronic hepatitis C (CHC). The present study
examined the oxidative status in plasma of patients with CHC who received pegylated interferon
and ribavirin therapy. The following groups were included: (1) sustained virological response
(28 patients), (2) null response (26 patients), (3) breakthrough (24 patients), (4) relapse
(24 patients), (5) spontaneous cure (23 patients) and (6) twenty five normal subjects as a control
group. Markers of oxidative stress including plasma malondialdehyde, nitric oxide, reduced
glutathione, total antioxidant capacity and uric acid as well as serum ALT, AST, alkaline
phosphatase, total bilirubin, albumin, prothrombin time were studied. The study indicated
significant decline in reduced glutathione and total antioxidant capacity and markedly elevated
levels of malondialdehyde and nitric oxide in all groups compared with the controls. Null
response group had the highest levels of malondialdehyde and nitric oxide. Nitric oxide was
significantly higher in those with null response compared with all other groups and with control
subjects. Uric acid was significantly higher in spontaneous cure group compared with all
other groups and with the controls. We concluded that CHC patients had increased oxidative
stress. The oxidative status in plasma of these patients was not changed by antiviral therapy.
The study also showed an important contribution of nitric oxide in null response patients.
High serum uric acid did not interfere with the response and/or did not predict the response to
antiviral therapy.
Keywords: chronic hepatitis C, interferon, oxidative stress

TGF-β1 is a cytokine with many different effects on cell proliferation,
differentiation and inflammation and can protect against the development of
COPD. This work aims to study the association between COPD and the TGF-β1
gene genotypes.
Material and methods: The study included 70 males: 25 smokers with COPD,
25 resistant smokers, and 20 normal non-smokers as the control. They were
subjected to spirometry pre- and post-bronchodilator (FEV1, FEV1/FVC), estimation
of serum level of TGF-β1 gene by PCR and RFLP.
Results: The percent of Pro-Leu was 28% in the COPD group, 84% in the resistant
smokers group and 85% in the control group. There was a highly significant
statistical difference in FEV1% of predicted associated with the distribution
of TGF-β1 gene genotypes: 56.9 ±8.4% with Pro-Leu genotype and 35.5 ±8.8%
with Leu-Leu genotype in COPD patients, 93.2 ±6.2% with Pro-Leu genotype
and 86.7 ±0.9% with Leu-Leu genotype in the resistant smokers group.
Conclusions: The Pro-allele genotype is associated with increased production
of TGF-β1, which has a protective role against the development of COPD and is
important in preserving the decline of FEV1 in COPD patients.
Key words: chronic obstructive pulmonary disease, leucine, proline domains.