Plasmid-mediated antibiotic resistance among uropathogens in primigravid women-Hyderabad, India.

With the growing threat of antimicrobial resistance worldwide, uncovering the molecular epidemiology is critical for understanding what is driving this crisis. We aimed to evaluate the prevalence of plasmid-mediated-quinolone-resistance (PMQR) and extended-spectrum beta-lactamase- (ESBL) producing gram-negative organisms among primigravid women with bacteriuria. We collected urine specimens from primigravid women attending their first antenatal visit at Gandhi Hospital during October 1, 2015 to September 30, 2016. We determined antimicrobial susceptibility and ESBL and quinolone resistance using VITEK-2. We performed polymerase chain reaction amplification on resistant isolates for detection of ESBL-encoding genes (TEM, SHV, CTX-M) and PMQR genes (qnrA, qnrB, qnrD, qnrS, aac (6')-Ib-cr). Of 1,841 urine samples, 133 demonstrated significant bacterial growth with gram-negative bacilli accounting for 85% of isolates, including Escherichia coli (n = 79), Klebsiella pneumoniae (n = 29), Sphingomonas (n = 3), Enterobacter (n = 1), and Citrobacter (n = 1). We found 65% of E. coli isolates and 41% of K. pneumoniae isolates were ESBL positive. Of ESBL-positive isolates, the most common genes conferring resistance were TEM-1 (66.7%) followed by CTX-M-15 (33.3%). Fifty-seven percent of ESBL-positive E. coli also demonstrated resistance to quinolones with the most common PMQR genes being qnr-S (62.5%) and aac (6')-Ib-cr (37.5%). We did not find any resistance to quinolones among ESBL-positive K. pneumoniae isolates. Across different classes of antibiotics we found a strong clustering of multi-drug resistance in E. coli with over 45% of ESBL-positive isolates demonstrating resistance to at least three classes of antibiotics. This study emphasizes the high prevalence of plasmid-mediated ESBL and quinolone resistance in community-acquired urinary tract infections of primigravid women. The overall abundance of multi-drug-resistant isolates in this population is alarming and may present therapeutic challenges.

Risk factors for endophthalmitis following cataract surgery-our experience at a tertiary eye care centre in India.

AIM: To determine the risk factors for acute endophthalmitis after cataract extraction in a tertiary care centre in India. METHODS: We performed a nested case control study within a retrospective cohort. The surgical records of all patients with clinically diagnosed endophthalmitis within one month after cataract surgery, performed between January 2006 and December 2009, were reviewed. These were compared with randomly selected age and gender-matched controls, from patients having routine cataract surgery within ±1wk of the endophthalmitis case. Univariable and multivariable analysis were performed to identify risk factors for endophthalmitis. RESULTS: Of the total 33 856 cataract surgeries performed during this period, there were 57 cases of postoperative acute endophthalmitis that met our study criteria. Thus, the overall incidence of endophthalmitis in our cohort was 1.6 per 1000 cataract extractions performed. Mean age of cases was 55.9y (SD: 10.9y) and for controls was 55.6y (SD: 9.8y). Thirty-five cases (61.4%) and 133 controls (59.6%) were males. Median time of onset of endophthalmitis was 4d (IQR 2-9d; range: 1-30d). Thirty-nine cases (68.4%) presented within 7d and 27 cases (47.4%) were culture positive. Two hundred and twenty-three age and gender matched controls were selected. In multivariate analysis, endophthalmitis was associated with posterior capsular rupture (PCR) during surgery (OR 6.98, 95%CI: 2.22-21.98), phacoemulsification via scleral incision with a foldable intraocular lens (IOL) implantation (OR 3.02, 95%CI: 1.13-8.04) and ocular co-morbidity (OR 2.32, 95%CI: 1.11-4.87). CONCLUSION: PCR, presence of ocular co-morbidity, and phacoemulsification via scleral incision with foldable-IOL were found to be independent risk factors for acute endophthalmitis.

Role of Human papilloma virus infection and altered methylation of specific genes in esophageal cancer.

BACKGROUND: Evaluation of Human papilloma virus (HPV) and its association with promoter methylation of candidate genes, p53 and Aurora A in esophageal cancer. MATERIALS AND METHODS: One hundred forty-one esophageal tissue samples from different pathologies were evaluated for HPV infection by PCR, while the promoter methylation status of p53 and Aurora A was assessed by methylation-specific restriction based PCR assay. Statistical analyses were performed with MedCalc and MDR software. RESULTS: Based on endoscopy and histopathology, samples were categorized: cancers (n=56), precancers (n=7), esophagitis (n=19) and normals (n=59). HPV infection was found to be less common in cancers (19.6%), whereas its prevalence was relatively high in precancers (71.4%), esophagitis (57.8%) and normals (45.7%). p53 promoter methylation did not show any significant difference between cancer and normal tissues, whereas Aurora A promoter methylation demonstrated significant association with disease (p=0.00016, OR:5.6452, 95%CI:2.18 to 14.6) when compared to normals. Aurora A methylation and HPV infection was found in a higher percentages of precancer (66.6%), esophagitis (54.5%) and normal (45.2%) when compared to cancers (14.2%). CONCLUSIONS: Aurora A promoter methylation is significantly associated with esophageal cancer, but the effect of HPV infection on this epigenetic alteration is not significant. However MDR analysis showed that the hypostatic effect of HPV was nullified when the cases had Aurora methylation and tobacco exposure. Further HPV sub-typing may give an insight into its reduced prevalence in esophageal cancer verses normal tissue. However, with the present data it is difficult to assign any significant role to HPV in the etiopathology of esophageal cancer.

Phototherapeutic keratectomy versus alcohol epitheliectomy with mechanical debridement for superficial variant of granular dystrophy: a paired eye comparison.

PURPOSE: To compare outcome of phototherapeutic keratectomy (PTK) and alcohol epitheliectomy with mechanical debridement for superficial variant of granular dystrophy. METHOD: Case report. A 7 year old girl with superficial variant of granular dystrophy with an visual acuity of 20/70 both eyes, underwent a simultaneous PTK in right eye (OD) and alcohol epitheliectomy with mechanical debridement in left eye (OS). Post operatively the visual acuity, refraction and corneal clarity were assessed. RESULTS: On the first post-operative day uncorrected visual acuity was 20/50 in both eyes. At 2 weeks, vision improved to 20/40 with +2.50 -2.00×180 in OD and +1.50 -1.50×180 in OS. At 2 years, vision and refraction were 20/25 with +2.50 -2.00×180 in OD and 20/25 with +1.25 -1.50×170 OS respectively. There was early recurrence of the granular dystrophy. CONCLUSION: The visual outcomes of both PTK and alcohol epitheliectomy are similar for superficial variant of granular dystrophy. The hyperopic shift with PTK is a matter of concern. Alcohol epitheliectomy can be used in treatment of superficial corneal dystrophies where PTK is unavailable.

No evidence for the role of somatic mutations and promoter hypermethylation of FH gene in the tumorigenesis of nonsyndromic uterine leiomyomas.

Fumarate hydratase (FH) gene is reported to have specific involvement in syndromic uterine tumors, but its role in nonsyndromic forms is still unclear. Hence, the present study has aimed to screen the role of promoter methylation status and mutations in exon 2 and 7 regions of FH gene in the genesis of nonsyndromic uterine leiomyomas. Leiomyoma and myometrium tissues were collected from 85 hysterectomized uterine specimens. DNA from each of the biopsy was subjected to PCR, methylation-specific restriction assay, and DNA sequencing. In silico analysis was carried out to identify the impact of sequence variants on the protein structure. Chi-square (χ (2)) test was used to compare the promoter methylation proportions of leiomyoma and myometrium tissues. No sequence variants were observed in exon 2 region, but three novel heterozygous germ line sequence variants, i.e., c.1010A > C, c.1021 G > A, and c.1066 T > C in exon 7 region of the FH gene were detected in 14/85 (16.5 %) of the cases examined. In silico analysis results showed that c.1010A > C and c.1021 G > A mutations damage the structure and function of FH, whereas c.1066 T > C mutation is mostly tolerant or neutral. No significant difference of FH promoter methylation status between the leiomyoma (11.76 %) and myometrium (5.88 %) tissues was observed (P = 0.176). Therefore, it is concluded that somatic mutations in FH do not show pronounced effect in nonsyndromic uterine leiomyomas compared to that of their syndromic counterparts. However, higher frequency of FH mutations in leiomyoma cases raises the need to conduct larger number of prospective case-control and family-based studies to assess them as risk markers to nonsyndromic leiomyomas.

Molecular pathogenesis of endometriosis; Toll-like receptor-4 A896G (D299G) polymorphism: a novel explanation.

BACKGROUND: Endometriosis is a distressing gynecological disorder. Toll-like receptor 4 (TLR4) is specific for recognition of the molecular pattern of gram-negative bacteria. TLR4 is present on the surface of endometrial cells. Their role in the molecular pathogenesis of endometriosis is postulated through a novel explanation. OBJECTIVE: To study the TLR4 A896G polymorphism in cases of endometriosis and age- and sex-matched healthy controls and evaluate its role in the molecular pathogenesis of endometriosis. DESIGN: Case-control study, involving patients from four gynecological centers from Hyderabad Samples: The study was carried out on 400 women who include 200 surgically confirmed cases of endometriosis and 200 healthy women volunteers in whom endometriosis was excluded by the standard diagnostic criteria. RESULTS: An association of G allele, GG, and AG genotype of TLR4 A896G polymorphism was seen in cases of endometriosis. "G" allele was found to be significantly associated with endometriosis (odds ratio=4.4827; 95% confidence interval: 2.2829-8.8021; χ(2) p<0.0001). An eightfold increase of endometriosis risk was seen in women who carry GG genotype, whereas AA genotype can be considered as protective in our population. CONCLUSION: TLR4 A896G polymorphism (rs4986790) is a functional polymorphism resulting in hypo-responsiveness of the receptor, thus resulting in peritoneal inflammation. The molecular microenvironment becomes favorable for the endometrial cells to adhere to the peritonium, thereby resulting in the initiation of endometriosis.