Zahid Shakoor

Cytokine gene polymorphisms and serum cytokine levels in patients with idiopathic pulmonary fibrosis

BackgroundStudies have demonstrated associations between cytokine gene polymorphisms and the risk of idiopathic pulmonary fibrosis (IPF). We therefore examined polymorphisms in the genes encoding interleukin (IL)-6, IL-10, interferon gamma (IFN-γ), tumor necrosis factor alpha (TNF-α), and transforming growth factor-beta 1 (TGF-β1), and compared the serum levels of these cytokines in IPF patients and healthy controls. Furthermore, we examined the association of the studied genotypes and serum cytokine levels with physiological parameters and the extent of parenchymal involvement determined by high-resolution computed tomography (HRCT).MethodsSixty patients with IPF and 150 healthy controls were included. Cytokine genotyping was performed using the polymerase chain reaction sequence specific primer (PCR-SSP) method. In a subset of patients and controls, serum cytokine levels were determined by enzyme-linked immunosorbent assay.ResultsThere was no difference between IPF patients and controls in the genotype and allele distributions of polymorphisms in TNF-α, IFN-γ, IL-6, IL-10, and TGF-β1 (all pu2009>u20090.05). The TNF-α (−308) GG, IL-6 (−174) GG and CG, and IL-10 (−1082, -819, -592) ACC ATA genotypes were significantly associated with HRCT scores (all pu2009<u20090.05). IL-10 (−1082, -819, -592) ACC haplotype was associated with the diffusion capacity of the lung for carbon monoxide, and ATA haplotype was associated with the partial pressure of oxygen (PaO2) (all pu2009<u20090.05). The TGF-β1 (codons 10 and 25) TC GG, TC GC, CC GG and CC GC genotypes were significantly associated with the PaO2 and HRCT scores (pu2009<u20090.05). The TGF-β1 (codons 10 and 25) CC GG genotype (5 patients) was significantly associated with higher PaO2 value and less parenchymal involvement (i.e., a lower total extent score) compared to the other TGF-β1 genotypes (81.5u2009±u200911.8xa0mm Hg vs. 67.4u2009±u200911.1xa0mm Hg, pu2009=u20090.009 and 5.60u2009±u20091.3 vs. 8.51u2009±u20092.9, pu2009=u20090.037, respectively). Significant differences were noted between patients (nu2009=u200938) and controls (nu2009=u200936) in the serum levels of IL-6 and IL-10 (both, pu2009<u20090.0001), but not in the levels of TNF-α and TGF-β1 (both, pu2009>u20090.05).ConclusionThe studied genotypes and alleles do not predispose to the development of IPF but appear to play an important role in disease severity. Our results suggest that the TGF-β1 (codons 10 and 25) CC GG genotype could be a useful genetic marker for identifying a subset of IPF patients with a favorable prognosis; however, validation in a larger sample is required.

ESBL-producing Escherichia coli and Klebsiella pneumoniae at a tertiary care hospital in Saudi Arabia

INTRODUCTIONnThe increasing frequency and antibiotic resistance among extended-spectrum β-lactamases (ESBLs)-producing bacteria are posing a serious threat. This study sought to investigate the frequency and antibiotic susceptibility of ESBL-producing E. coli and K. pneumoniae at a tertiary care hospital.nnnMETHODOLOGYnData were collected from samples sent to the microbiology laboratory between 2006 and 2010 at King Khalid University Hospital, Riyadh. ESBLs were confirmed using Etest strips of cefotaxime/cefotaxime + clavulanic acid, ceftazidime/ceftazidime + clavulanic acid, and cefepime/cefepime + clavulanate.nnnRESULTSnOut of 17,105 samples, 1,076 (6.3%) ESBL-producing isolates of E. coli (808) and K. pneumoniae (268) were confirmed. Among these, 680 (63.2%) isolates were found in urine samples, followed by 287 (26.7%) in superficial swabs, deep wounds swabs, tissues and sterile body fluids, 71 (6.6%) in respiratory, and 38 (3.5%) in blood samples. The overall frequency rates of ESBL E. coli and K. pneumoniae were 6.6% and 5.5%, respectively. The frequency of ESBL-producing E. coli and K. pneumoniae increased significantly during the study period. E. coli resistance against cotrimoxazole was 71.1%, followed by ciprofloxacin (68.2%) and gentamicin (47%). Similarly, 62.7% of K. pneumoniae isolates were resistant to gentamicin, 59.5% to cotrimoxazole, and 49.8% to ciprofloxacin. There was no statistically significant change in antimicrobial resistance over the study period.nnnCONCLUSIONSnAlthough the frequency rates of ESBL-producing E. coli and K. pneumoniae increased, no change in the anti-microbial susceptibility was observed over the study period.

Combined effect of a mixture of tetracycline, acid, and detergent, and Nisin against Enterococcus faecalis and Actinomyces viscosus biofilms

Objectives: To evaluate the combined effect of a mixture of tetracycline, acid, and detergent (MTAD) and Nisin against Enterococcus faecalis (E. faecalis) and Actinomyces viscosus (A. viscosus) biofilms. Methods: This study was conducted between June and December 2013 in collaboration with Dental Caries Research Chair, College of Dentistry, King Saud University, Riyadh, Saudi Arabia. Single-species biofilms (n=9/species/observation period) were generated on membrane filter discs and subjected to 5, 10, or 15 minute incubation with MTADN (MTAD with 3% Nisin), 5.25% sodium hypochlorite (NaOCl), or normal saline. The colony forming units were counted using the Dark field colony counter. Results: A 100% bactericidal effect of 5.25% NaOCl was noted during the 3 observation periods; a significant reduction (p=0.000) in mean survival rates of E. faecalis (77.3+13.6) and A. viscosus (39.6+12.6) was noted after 5 minutes exposure to MTADN compared with normal saline (78000000+5291503) declining to almost no growth after 10 and 15 minutes. The survival rates of the E. faecalis and A. viscosus biofilm were no different after treatment with MTADN and 5.25% NaOCl at the 3 observation periods (p=1.000). Conclusion: A combination of MTAD and Nisin was as effective as NaOCl against E. faecalis and A. viscosus biofilms.

Performance of the BD ProbeTec ET direct detection assay for the analysis of Mycobacterium tuberculosis in respiratory and non-respiratory clinical specimens

Objectives Early detection of Mycobacterial tuberculosis infection (MTB) is pivotal for the treatment of tuberculosis (TB). Background This study was performed to evaluate the performance of BD ProbeTec ET direct detection assay (DTB) against the gold standard culture technique for confirmation of MTB infection. Methods A total of 266 consecutive and non-duplicate clinical specimens for detection of MTB were included in this study. There were 118 respiratory and 148 non-respiratory samples. All samples were tested by microscopy for acid-fast bacillus (AFB), MTB culture and biochemical identification with simultaneous testing by DTB. Results A total of 88 samples (33%) were culture-positive for MTB including 39/118 respiratory, 29/99 fluid and 20/49 tissue samples. DTB sensitivity for respiratory samples was 97% and specificity was 96% with a positive predictive value (PPV) of 93% and negative predictive value (NPV) of 99%. Sensitivity of DTB in fluid samples was 80%, specificity 88%, PPV 69% and NPV 93% whereas sensitivity of DTB for tissue samples was 25%, specificity 90%, PPV 63% and NPV 63%. Of the 50 (56.8%) smear-positive samples, DTB sensitivity was 100% for respiratory, 85% for fluid and 100% for tissue samples. Conclusion DTB performed within acceptable limits for the rapid detection of MTB in respiratory samples compared to fluid and tissue specimens.

IRAK2 is associated with susceptibility to rheumatoid arthritis

This study was performed to investigate the association of the single nucleotide polymorphisms of interleukin-1 receptor-associated kinase 2 (IRAK2) rs3844283 and rs708035 with rheumatoid arthritis (RA). IRAK2 rs3844283 and rs708035genotyping was determined by mutagenically separated PCR with specifically designed primers in a cohort of 222 (30 men, 192 women, mean age 49xa0years) adult RA patients and 224 matched controls. IRAK2 rs3844283 C allele was detected in 66% of RA patients and 74% of controls. The CC genotype was the most frequent genotype in both RA patients (45.5%) and the controls (56.3%). The G allele was found to be associated with RA susceptibility (ORxa0=xa01.47, 95% CIxa0=xa01.10–1.96, pxa0=xa00.008). The GG genotype was found to be associated with RA in the co-dominant and the dominant models (ORxa0=xa02.03, 95% CIxa0=xa01.08–3.81, pxa0=xa00.042 and ORxa0=xa01.54, 95% CIxa0=xa01.06–2.23, pxa0=xa00.023, respectively). IRAK2 rs708035 was found not to be in the Hardy-Weinberg equilibrium. The hyperfunctional IRAK2 rs708035 A allele was more frequent in RA patients than in controls (69.9 versus 62.2%, respectively, pxa0=xa00.015). Moreover, IRAK2 rs708035 and IRAK2 rs3844283 were in linkage disequilibrium and the GA haplotype was significantly more frequent in RA patients than in controls (pxa0=xa00.034). This study for the first time ever reports the association of IRAK2 rs3844283, IRAK2 rs708035, and the corresponding haplotypes with RA. Functional studies are recommended to elucidate the risk posed by the GA haplotype for the development of RA.

Screening for skin-sensitizing allergens among patients with clinically suspected allergic contact dermatitis

Objectives: To detect common skin-sensitizing agents among patients experiencing allergic contact dermatitis (ACD). Methods: This was a retrospective study of 152 patients with clinically suspected ACD who underwent patch testing in an allergy clinic at King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia between January 2012 and February 2015. Of these patients, only 74 (48%) patients reacted to one or more contact allergens. This group of patients included 58 (78.4%) women and 16 (21.6%) men (mean age: 37.8±13.8 years). Patch testing was performed using the thin-layer rapid-use epicutaneous patch test panels. Results: Nickel sulfate was the most common sensitizing agent, with 26 (35.1%) patients yielding a positive result; followed by p-phenylenediamine in 17 (22.9%), butyl-tetra-phenol formaldehyde in 12 (16.2%), gold sodium thiosulfate in 10 (13.5%), and thimerosal in 6 (8.1%) patients. Nickel reactivity was significantly higher among women (41.4%) than among men (12.5%) (p<0.0001). Similarly, gold reactivity among women (15.5%) was also higher than among men (6.2%) (p≤0.02). Conclusion: The high level of skin sensitization due to nickel, PPD, and gold in patients with ACD emphasizes the need for measures to decrease exposure to these sensitizing agents.

Neonatal brucellosis: A case report

Although brucellosis is not uncommon in Saudi Arabia, neonatal brucellosis has been infrequently reported. In this case of neonatal brucellosis, Brucella abortus was isolated by blood culture from both the mother and the neonate. Serology was positive only in the mother.

Acute back pain: A survey of primary health care physicians' awareness and knowledge of "red flag" signs

Background: The failure to detect red flag signs in patients presenting with acute low back pain can adversely affect the outcome of management. This can seriously affect the quality of life and productivity of the patient. Objective: The present questionnaire-based study was performed to assess the knowledge and awareness of red flag signs among primary health care physicians managing patients with acute back pain in Riyadh, Saudi Arabia. Materials and Methods: The study sample size was comprised of 80 subjects. The level of knowledge was assessed by means of a new structured self-administered questionnaire. The design of this questionnaire was based on the Agency for Health Care Research and Policy (AHCRP) guidelines for detection of red flag signs. Physicians were asked about red flag signs that indicate the presence of tumor, infection, spinal fracture, or cauda equina syndrome. Results: Sixty-eight (85% of total) physicians were aware of red flag signs. Of the 68 physicians who were aware of the red flag signs, 58 (72%) were aware of neurological deficit, 36 (45% of total) were aware of extremes of age (<10 years and >50 years), and 33 (41% of total) were aware of and routinely inquired about the history of spinal trauma, whereas only 24 (30% of total) were aware of and inquired about constitutional symptoms in their patients with acute back pain. Conclusion: Although low back pain is extremely common, knowledge and awareness of red flag signs of primary health care physicians managing patients with acute back pain in Riyadh appear to be inadequate. This indicates a lack of adherence to the international guidelines. Specific educational programs should target these deficiencies and increase awareness.

Assessment of antigen presenting cell infiltration in lung tissues of patients with bronchiectasis

Background: Bronchiectasis is a chronic disease characterized by permanent dilatation of the conducting airways accompanied by sustained inflammation. Aims: To assess whether chronic inflammation of lungs in bronchiectasis is associated with alterations in the numbers of infiltrating antigen presenting cell (APC). Setting and Design: Lobectomy specimens from 12 nonsmoker, nonasthmatic patients with acquired (noncongenital) bronchiectasis and six control patients were included in the study. Histopathology slides were reviewed, and immunohistochemical markers for dendritic cells (DCs) macrophages and Langerhans cells have been applied and analyzed. Materials and Methods: Tissue specimens were stained by immunohistochemistry using markers for DCs (CD83 and CD23), macrophages (CD68 and CD163), and Langerhans cells (CD1A and S-100 protein). The mean cell counts of stained cells in five high power microscopic fields were recorded. Statistical Analysis Used: Descriptive statistics, mean, standard deviation, median, and interquartile range were used. A nonparametric Mann-Whitney U-test was used to compare cell counts between bronchiectasis and control patients. P <0.05 was considered significant. Results: The mean age of patients with bronchiectasis and controls was 36.7 ± 16.6 and 31.8 ± 22.6 years, respectively. The predominant cell type among the patients was macrophage (median 50.5) followed by DCs (median 44.85), histiocytes (median 32), and Langerhans cells (median 5%). Compared to the controls a significantly higher number of macrophages (P = 0.01), DCs (P = 0.001), and Langerhans cells (P = 0.014) were present. Conclusion: Chronic inflammatory response in acquired (noncongenital) bronchiectasis is most probably mediated by increased infiltration of APCs in lung tissues.