Study of Neutrophil Gelatinase Associated Lipocalin and Kidney Injury Molecule-1 in Patients with Type 2 Diabetes Mellitus Nephropathy.

BACKGROUND: There is a need for accurate and rapid biomarkers for the early diagnosis of diabetic nephropathy (DN). We aimed to study the accuracy of urinary neutrophil gelatinase-associated lipocalin (uNGAL), urinary kidney injury molecule-1 (uKIM-1), and blood NGAL (bNGAL) in type 2 diabetics as biomarkers for diagnosis of DN. METHODS: The study was a retrospective case-control study that included 30 control subjects, 40 diabetics with normo-albuminuria < 30 mg/g and eGFR > 60 mL/minute/1.73 m2, and 30 diabetics with albuminuria > 30mg/g and eGFR < 60mL/minute/1.73 m2. Blood and urine samples were obtained to determine levels of bNGAL, uNAGAL, and uKIM1. RESULTS: There was a significant increase in bNGAL, uNGAL, uKIM 1, uNGAL/creatinine and uKIM 1/creatinine among diabetics with albuminuria compared to diabetics with normoalbuminuria and normal control (p < 0.001 for all markers). For diagnosis of early DN, both bNGAL and uKIM 1 had sensitivity and specificity of 100% for each at cutoff values of 322.5 pg/mL and 74.25 ng/mL, respectively. uNGAL had a sensitivity of 97.5% and a spec-ificity of 100% at a cutoff point of 565 ng/mL. uKIM1/creatinine at a cutoff of 51.2 had a sensitivity of 100% and specificity of 100%. CONCLUSIONS: The present study highlights the accuracy of urinary KIM1 and NGAL and blood NGAL as biomarkers for the diagnosis of nephropathy in the early stage of diabetic nephropathy. There were positive correlations with kidney function tests creatinine, blood urea nitrogen, and the presence of albuminuria.

Serum interleukin 33 levels and single nucleotide polymorphism rs1929992 in Egyptian patients with chronic asthma.

Bronchial asthma is a common chronic inflammatory disease affecting the airway. Cytokines have a pivotal role in regulation of the immune response, and in development of asthma. Interleukin 33 is a newly discovered member of cytokines, belongs to interleukin 1 family. Previous studies have reported that expression of IL33 is associated with bronchial asthma. This study aimed to evaluate the prevalence of interleukin 33 (IL33) single nucleotide polymorphism (SNP) rs1929992 in asthmatic patients and determine the relation of IL33SNP to IL33 serum level. The Results of RFLP were validated by using sterile distilled water. This study included 100 patients from Egypt, Beni Suef governorate (Upper Egypt) and Mansoura governorate (Delta region), complaining of chronic asthma and 100 control subjects with matched sex and residence. Blood samples from study subjects were used for determination of serum IL33by ELISA and IL33 SNP rs1929992 by PCR-RFLP. There was no significant difference between the proportions of IL33 SNP rs1929992 genotypes in asthma patients and the control group. Allele 'A' predominates in asthmatics though this did not achieve statistical significance (P=0.071). IL33 level was compared in the three IL33 SNP rs1929992 genotypes; G/G, G/A, and A/A, and it revealed no significant difference (P = 0.958). The association between IL33 with asthma showed that the log-additive model is the best inheritance model which marks allele 'A' as the risk allele. In contrast, IL33 serum level was significantly higher in severe asthma than the moderate asthma and the mild type (P<0.0005). Spearman's correlation test showed that IL33 level rises as asthma severity increases (rs=0.880, P<0.0005). In conclusion our data revealed no evidence that SNP of IL33 rs1929992 may contribute to the development of asthma in Egyptian population. However, there is a strong positive correlation between IL33 serum level and asthma severity.

Can Triggering Receptors Expressed on Myeloid Cells be used for Diagnosis of Salmonella Typhi Infection?

BACKGROUND: There is a need for rapid and accurate diagnostic biomarker for diagnosis of Salmonella fever. AIMS: The aim of the present study was to assess the importance of procalcitonin (PCT), Soluble Triggering Receptors expressed on Myeloid Cells 1 (sTREM1) and C- reactive protein (CRP) in the diagnosis of enteric fever with positive blood culture for S.typhi. METHODS: Blood samples were withdrawn from 200 patients with suspected enteric fever and subjected for the determination of CRP, PCT and sTREM-1. RESULTS: The sensitivity and specificity for PCT cut off were 97.7% & 82.5%, for CRP the sensitivity and specificity were 95.3% and 77% and for s-TREM-1 the sensitivity and specificity were 95.3% & 77%. CONCLUSION: S-TREM-1 may be considered as a novel biomarker for the diagnosis of enteric fever with good sensitivity and specificity.

Molecular Characterization of ß-Lactam Resistant Streptococcus pneumoniae.

BACKGROUND: Streptococcus pneumoniae (S. pneumoniae) is a commensal bacterium that normally colonizes the human nasopharyngeal cavity. Once disseminated, it can cause several diseases, ranging from non-invasive infections such as acute otitis media and sinusitis through to invasive infections with higher mortality. Antibiotic resistance among S. pneumoniae has increased dramatically and penicillin-resistant strains have spread worldwide with pneumococcus also being resistant to other types of antibiotics like erythromycin, tetracycline, and chloram-phenicol. The aim of the present study was to study the susceptibility of the isolated strains to ß-lactam and other antibiotics from different classes and to determine the prevalence of ß-lactam resistance genes among S. pneumoniae clinical isolates. METHODS: From a total of 178 sputum samples, isolates identified by standard microbiological method as S. pneu-moniae were subjected to antibiotic susceptibility tests to ß-lactam and non ß-lactam antimicrobial agents by disk diffusion method. Biofilm formation was detected by microtitration plate and the resistance genotype was also determined using multiplex PCR technique with primers designed for PBP genes. RESULTS: Out of 178 sputum samples, sixty isolates were recovered as Streptococcus pneumoniae. Most of isolates were multidrug-resistant (MDR) possessing a high (> 0.2) multiple antibiotic resistance index (MAR) value. Biofilm formation ability of isolates were strong, moderate, weak, and none, accounting for 21.67%, 45%, 25%, and 8.33% biofilm formers, respectively, and it was found that pbp1a, pbp2b, and pbp2x were present in 33 (55%), 25 (41.7%), and 45 (75%) of isolates, respectively. CONCLUSIONS: Streptococcus pneumoniae clinical isolates have an alteration in PBP resistance genes in response to ß-lactam therapy which subsequently lead to increased MDR phenomena among these clinically important pathogens. These findings necessitate continuous monitoring of antimicrobial resistance to guide the empirical treatment of pneumococcal disease, as well as to encourage reflections to support public immunizations strategies.

Clinicopathological study of occult hepatitis B virus infection in hepatitis C virus-associated hepatocellular carcinoma.

BACKGROUND: Occult hepatitis B virus infection (OBI) frequently occurs in patients with chronic hepatitis C (CHC) infection, but the influence of OBI on CHC outcome is still uncertain. The aim of the present study was to clarify the clinical and pathological characteristics of OBI in CHC-related hepatocellular carcinoma (HCC). PATIENTS AND METHODS: DNA was obtained from serum and tumor tissue of patients with hepatitis C virus (HCV)-related HCC with negative HBsAg and from patients with HCV-related liver cirrhosis. HBV-DNA was detected using qPCR. Clinicopathological features were compared between patients with HCC with and without OBI. RESULTS: On the basis of positive serum and tissue HBV-DNA typing, the overall frequency of OBI was 50% in patients with HCV-related HCC. HBV genotype D was the most dominant, constituting 35.3% of HCC cases. Almost 80% of patients with OBI had anti-HBc, whereas 20% of patients had no serological markers. Tissue HBV-DNA showed significant association with positive serum HBV-DNA, anti-HBc, and genotype D. There were no clinical differences between patients with HCC with and without OBI; however, patients with OBI tended to be younger. HCC cases with positive OBI were significantly associated with positive anti-HBc antibodies and late histological grades (3-4). Multivariate logistic regression analysis revealed that the presence of OBI was a predictor of more advanced HCC histological grades in patients with HCV infection. CONCLUSION: OBI was detected in 50% of HCV-infected patients with HCC. OBI was strongly associated with the presence of anti-HBc antibodies. Patients with HCC with positive OBI were younger and had more advanced HCC histological grades.

Study of MicroRNA-122 as a Diagnostic Biomarker of Sepsis.

Sepsis in intensive care units (ICUs) represent a threat with need for rapid and accurate diagnosis. We aimed to assess miR-122 as an early biomarker for diagnosis and outcome prediction in patients with hospital acquired sepsis in ICU. This case control study included 25 adults' patients with sepsis and 25 patients with local wound infections as a control group. C-reactive protein (CRP), total leucocytes count (TLC), liver function, and molecular determination of miR-122 levels were assessed. miR-122 had significant higher area under curve (AUC) when compared with CRP and TLC for differentiation of sepsis from wound infections. The cut off value for miR-122 was 0.16 folds expression with sensitivity, specificity and accuracy 100%. The TLC cut off value was 14.00 x103/cmm with 100% sensitivity, 84% specificity and 92% accuracy. While CRP cut off value was 41 mg/l with 76.0% sensitivity, 100% specificity, and 88.0% accuracy. Multivariate logistic analysis revealed non statistically significant difference between survivors and non survivors regarding sepsis biomarkers. Receiver operation curve (ROC) for different biomarkers, CRP, TLC and miR-122 to differentiate patients with poor outcome of sepsis compared to patients with recovery, revealed that AUC was 0.61, 0.6, and 0.45 respectively. miR-122 as a prognostic biomarker for sepsis had 66.6% sensitivity, 50% specificity, and 56.0% accuracy. The present study highlights important points in the use of biomarkers in diagnosis of sepsis in adults' patients above 50 years old. miR-122 is an accurate and specific biomarker for diagnosis of sepsis. miR-122 has limited predictive value for determination of the outcome of patients with sepsis even when used in combination with another biomarker such as CRP and TLC.

Epstein Barr Virus in Patients with Nephropathy Associated with Systemic Lupus Erythematous, Pilot Study in Egyptian Patients.

Systemic lupus erythematosus (SLE) is an autoimmune disease affecting young age adults especially females. Infection with Epstein Barr virus (EBV) represents a common pathogen associated with SLE activity. This study investigates the occurrence of EBV in SLE patients with renal complications by serological markers and molecular detection of EBV genome in renal biopsies and examine the association of EBV with the pathological grades in renal diseases. The study included nineteen patients with systemic lupus nephropathy and thirteen patients with non-lupus nephropathy. Renal biopsies were subjected to detection of EBV by PCR. Serum autoantibodies (anti- dsDNA, anti-Sm and anti-RNP) and EBV-IgM and IgG antibodies were detected by ELISA. The commonest autoantibody was anti- dsDNA (73.7%) followed by anti-Sm (57.8%) and anti-RNP (31.6%). The EBV-PCR revealed that 31.6% of patients with lupus nephropathy showed positive LMP1 gene expression in renal biopsies On the other hand, serological markers for EBV showed no significant difference between both groups; IgM for EBV was positive in 26.3% of patients with lupus nephropathy and 7.7% in non-lupus nephropathy, while IgG was positive in 26.3% and 15.4 % respectively. Positive LMPI-PCR was demonstrated in all (3/3) patients with severe degree of nephropathy as compared to 23.1% of patients with moderate degree of nephropathy. A significant association was found between EBV-PCR and anti-Sm, (P=0.01), anti- dsDNA (P=0.001), and IgG for EBV and anti- dsDNA (P=0.03). In conclusion, Molecular detection of EBV DNA in renal biopsies can be applied for laboratory diagnosis in SLE nephropathy. The severity of nephropathy associated with SLE seems to be aggravated by the presence of EBV. Further extended studies are required to elucidate this association.

Kidney injury in infants and children with iron-deficiency anemia before and after iron treatment.

BACKGROUND: Our study aimed to investigate the effects of iron-deficiency anemia (IDA) on renal tubular functions before and after iron treatment for infants and children with IDA. We measured urinary levels of two kidney injury markers: neutrophil gelatinase-associated lipocalin (NGAL) and liver-type fatty acid-binding protein (L-FABP). MATERIAL AND METHODS: Thirty-six infants and children with IDA and 20 matched healthy controls were included. We assessed different laboratory parameters, estimated glomerular filtration rate, urinary levels of NGAL, and L-FABP. Urinary kidney injury markers were measured in IDA patients before and after 3 months of oral iron therapy. RESULTS: IDA patients had significantly higher urinary NGAL and L-FABP levels compared to their healthy controls. After 3 months of oral iron treatment, there was a significant improvement (decrease) in urinary NGAL and L-FABP in infants and children with IDA. Urinary markers returned to normal levels (healthy control levels) in children with IDA, but not for infants with IDA compared to their healthy controls. CONCLUSION: Subclinical kidney injury was found in infants and children with IDA. This injury was completely reversible in older children with IDA and partially reversible in infants with IDA after iron therapy. Higher urinary levels of kidney injury molecules in IDA infants after iron treatment are suggestive of more sensitivity of these infants to oxidative stress caused by iron therapy or may be due to the immaturity of the kidney and more damage caused by IDA which may require more time to recover.

Protective role of humoral immune responses during an outbreak of hepatitis E in Egypt.

Although the seroprevalence of hepatitis E virus (HEV) is approximately 80% in adult Egyptians living in rural areas, symptomatic HEV-caused acute viral hepatitis (AVH) is sporadic and relatively uncommon. To investigate the dichotomy between HEV infection and clinical AVH, HEV-specific immune responses in patients with symptomatic and asymptomatic HEV infection during a waterborne outbreak in Egypt were examined. Of 235 acute hepatitis patients in Assiut hospitals screened for HEV infection, 42 (17.9%) were acute hepatitis patients confirmed as HEV-caused AVH; 37 (88%) of the 42 patients were residents of rural areas, and 14 (33%) were from one village (Kom El-Mansoura). Another 200 contacts of AVH cases in this village were screened for HEV and 14 (7.0%), all of whom were family members of AVH cases, were asymptomatic HEV IgM-positive. HEV infections in this village peaked during the summer. Asymptomatic HEV seroconverters had significantly higher levels of epitope-specific neutralising (p=0.006) and high avidity (p=0.04) anti-HEV antibodies than the corresponding AVH cases. In conclusion, naturally acquired humoral immune responses appear to protect HEV-exposed subjects from AVH during an HEV outbreak in Egypt.

Polymerase chain reaction of pleural biopsy is a rapid and sensitive method for the diagnosis of tuberculous pleural effusion.

BACKGROUND: Tuberculous pleural effusion occurs in 30% of patients with tuberculosis (TB). Rapid diagnosis of a tuberculous pleural effusion would greatly facilitate the management of many patients. Polymerase chain reaction (PCR) has been used to detect Mycobacterium tuberculosis in pleural fluid with highly variable sensitivity. OBJECTIVE: To improve our laboratory diagnosis of tuberculous pleural effusion. METHODS: We applied PCR to detect DNA specific for M tuberculosis in 33 of the studied pleural biopsy specimens using an IS986-based primer that was specific for mycobacterium complex, and compared it to the results of pleural fluid and biopsy cultures performed on either Lowenstein-Jensen (LJ) medium or BACTEC 12B liquid medium (Becton Dickinson Microbiology Systems; Cockeysville, MD), Ziehl-Neelsen (ZN) staining, and histopathology in 45 patients with pleural effusion. RESULTS: Of the 45 patients with pleural effusion who were studied, 26 patients received diagnoses of tuberculous pleural effusion that had been confirmed by either culture and or histopathology, 10 patients received diagnoses of exudative effusion due to causes other than TB, and 9 patients received diagnoses of transudative effusion. Histopathology of the pleural biopsy specimen had a sensitivity of 53.8%. The sensitivity of the ZN staining of pleural fluid and biopsy specimens was 0.0% and 3.8%, respectively. The sensitivity of the culture on both BACTEC 12B liquid medium and LJ medium was higher in pleural biopsy specimens (92.3%) than in pleural fluid specimens (15.4%; p > 0.001). The improvements of the BACTEC culture system improved and shortened the detection time of M tuberculosis in pleural biopsy specimens. PCR of pleural biopsy specimens had 90% sensitivity and 100% specificity. The positive predictive value and the negative predictive value for pleural biopsy specimen cultures were 100% and 90.5% vs 100% and 86.7% for pleural biopsy specimen PCRs. CONCLUSION: The overall accuracy of PCR of pleural biopsy was similar to the results of pleural biopsy culture, however, PCR of the pleural biopsy was much faster in reaching diagnosis. PCR of pleural biopsy is a useful method when used in combination with the BACTEC culture system and histopathologic examination of pleural biopsy to reach a rapid diagnosis of tuberculous pleural effusion.