The research investigates the virulence factors of Pseudomonas aeruginosa (P. aeruginosa), a pathogen known for its ability to cause human infections by releasing various exoenzymes and virulence factors. Particularly relevant in ocular infections, where tissue degeneration can occur, even after bacterial growth has ceased due to the potential role of secreted proteins/enzymes. Clinical isolates of P. aeruginosa, both ocular (146) and non-ocular (54), were examined to determine the frequency and mechanism of virulence factors. Phenotypic characterization revealed the production of alginate, biofilm, phospholipase C, and alkaline protease, while genotypic testing using internal uniplex PCR identified the presence of Exo U, S, T, Y, and LasB genes. Results showed a significant prevalence of Exo U and Y genes in ocular isolates, a finding unique to Indian studies. Additionally, the study noted that ocular isolates often contained all four secretomes, suggesting a potential link between these factors and ocular infections. These findings contribute to understanding the pathogenesis of P. aeruginosa infections, particularly in ocular contexts, and highlights the importance of comprehensive virulence factor analysis in clinical settings.

According to the World Health Organization, infectious diseases, particularly those caused by multidrug-resistant bacteria (MDR), are projected to claim the lives of 15 million people by 2050. Septicemia carries a higher morbidity and mortality rate than infections caused by susceptible Pseudomonas aeruginosa, and MDR-mediated ocular infections can lead to impaired vision and blindness. To identify and develop a potential drug against MDR P. aeruginosa, we employed in silico reverse genetics-based target mining, drug prioritization, and evaluation. Rare Lipoprotein A (RlpA) was selected as the target protein, and its crystal structure was geometrically optimized. Molecular docking and virtual screening analyses revealed that RlpA exhibits strong binding affinity with 11 compounds. Among these, 3-chlorophthalic acid was evaluated, and subsequent in vitro assays demonstrated significant anti-Pseudomonas activity with negligible cytotoxicity. The compound was further evaluated against both drug-susceptible and MDR P. aeruginosa strains in vitro, with cytotoxicity assessed using an MTT assay. The study demonstrated that 3-chlorophthalic acid exhibits potent anti-Pseudomonas activity with minimal toxicity to host cells. Consequently, this compound emerges as a promising candidate against MDR P. aeruginosa, warranting further investigation.

Stenotrophomonas maltophilia (S. maltophilia) is an emerging Gram-negative bacteria causing wide range of infections among immunocompromised patients. The current study elucidates the draft genome sequence of this underrated bacterial pathogen causing life-threatening diseases for the immunocompromised patients.

Purpose To evaluate the efficacy of CXL in treating fungal keratitis as an adjuvant therapy. Methods Detailed clinical examination microbiological investigation was performed. Twenty fungal keratitis patients were recruited and randomized into two groups: group 1 (n= 11, standard antifungal), group 2 (n=9, corneal collagen crosslinking with standard antifungal). Corneal scraping and tear samples collected were subjected to real-time PCR targeting ITS, TLR analysis and cytokine analysis. Results The mean time for complete resolution of ulcer for group 2 was significantly shorter compared to group 1 and the final mean BCVA was better for group 2. Expression of IL-1 beta, IL-8, IFN-gamma significantly decreased immediately post CXL in group 2 patients. Significant downregulation of TLR 6, TLR-3, TLR-4 was observed 3-days post CXL compared to group 1 patients. Conclusion Adjuvant effect of CXL was significant in treating fungal keratitis compared to standalone antifungal treatment.

BACKGROUND:Toll-like receptors (TLRs) play a significant role based on innate immune mechanism during viral infection. TLR signaling mechanism designates to protect the cells from invading viruses. The expression of TLRs during dengue virus (DENV) infection not yet well explained. This study evaluates the TLR gene expression from DENV-infected patient's cornea.METHODS:Reverse transcriptase PCR was performed for the detection and genotyping of viral nucleic acid from corneal grafts and DENV-infected cell suspension. TLR expression studies were done on DENV-infected cornea by real-time RT2 Profiler PCR Array.RESULTS:The reverse transcriptase PCR and genotyping confirmed the presence of DENV-3. TLR expression studies revealed the upregulated expression of TLR4, TLR7, TLR9 and TLR10.CONCLUSION:Molecular testing of DENV reveals that serological positivity induces transmission of the virus through cornea and stimulates the expression of TLR4, TLR7, TLR9 and TLR10, which may lead to up-regulation of innate pro-inflammatory response in the cornea.

Frosted branch angiitis (FBA), a rare form of retinal vasculitis presenting as bilateral perivascular sheathing, resembling the appearance of frosted tree branches in winter, was first reported by Ito et al.1 in 1976, in a young immunocompetent boy. FBA predominantly affects healthy young patients, the youngest reported in an 11-month-old infant2 and oldest in a 42-year-old patient.3 Classical symptoms include sudden onset of blurred vision with floaters and photopsiae. Fundus examination shows widespread perivascular translucent sheathing affecting both arterioles and venules, more commonly latter. Fluorescein angiography shows late staining of vessels with no obstruction of blood flow. Electroretinogram shows reduced amplitude and visual fields show generalized constriction. Medline search did not show any case of frosted branch angiitis in a patient with typhoid fever.

The original publication of this article [1] was missing the below author that made contributions to the research and the published article:

Purpose To describe the incidence, risk factors, clinical presentation, causative organisms, and outcomes in patients with endophthalmitis following pars plana vitrectomy (20G and minimally invasive vitrectomy surgery (MIVS). Methods Of 111,876 vitrectomies (70,585 20-G 41,291 MIVS) performed, 45 cases developed acute-onset, postoperative endophthalmitis. Results The rate of culture positive and culture negative endophthalmitis was 0.021% (2.1/10,000 surgeries) and 0.019% (1.9/10,000 surgeries) overall, 0.031% (3.1/10,000 surgeries) and 0.025% (2.5/10,000 surgeries) in 20G, and 0.005% (0.5/10,000 surgeries) and 0.007% (0.7/10,000 surgeries) in the MIVS group respectively. Potential predisposing factors were as follows: diabetes, 46.7%; vitrectomy for vascular retinopathies, 44.4%; and vitrectomy combined with anterior segment surgeries, 35.5%. The culture proven rates were 53.3% overall, 55.0% for 20G and 40.0% for MIVS. The most common organism was Pseudomonas aeruginosa for 20G. Klebsiella and Staphylococcus aureus were isolated in the two culture positive cases in MIVS group. The follow-up period for the patients with endophthalmitis was 586.14 ± 825.15 days. Seven were lost to follow up beyond one week. Of the remaining 38, 13 (34.2%) cases had a favorable visual outcome (i.e., best-corrected visual acuity [BCVA] > 5/200) and 24 (63.2%) had unfavorable visual outcome (BCVA < 5/200). Group with culture test results negative had significantly better outcomes (P < 0.05) as compared to those with positive. Conclusions MIVS does not increase the risk of endophthalmitis. Outcomes are poor despite appropriate treatment, particularly in cases with culture results positive.

Background: Dengue fever is the most prevalent form of flavivirus infection in humans. We have investigated whether corneoscleral tissue of the donor affected by dengue virus (DENV) harbors the virus. Purpose: To identify the risk for viral transmission through corneal transplants in areas where DENV circulates. Methods: Excised corneoscleral tissue from a cadaver with a history of viral hemorrhagic fever was analyzed using reverse transcriptase-polymerase chain reaction for the presence of DENV and chikungunya virus (CHIV). Results: DENV was detected in RNA extracted from the donor corneoscleral rim. Further genotyping of the viral isolate from the virus-infected cell harvest revealed DENV type 3 as the causative agent. CHIV was not detected. Conclusions: The data presented in this study recommend the implementation of polymerase chain reaction for detection of DENV and CHIV to analyze excised corneoscleral tissue of a donor with viral hemorrhagic fever.

Purpose: To identify the risk factors associated with endophthalmitis after pars plana vitrectomy (PPV). Design: Retrospective case-control study. Participants: All eyes that presented with endophthalmitis after PPV within 6 weeks in a tertiary eye care center were evaluated. There were 36 cases with endophthalmitis and 93 controls without endophthalmitis. Methods: Other patients undergoing operation by the same surgeon on the same date and in the same operating room were included as controls. Univariate and multivariate regression analyses were performed to evaluate the risk factors. Main Outcome Measures: Incidence and risk factors for endophthalmitis after PPV. Results: In this study, 36 cases and 93 controls met the inclusion criteria. For endophthalmitis, 3 independent risk factors were identified: systemic immunosuppression (odds ratio [OR], 10.673; 95% confidence interval [CI], 1.114-102.292; P = 0.04), balanced salt solution (BSS) or Ringer's lactate (RL) as vitreous substitute (OR, 5.288; 95% CI, 1.769-15.813; P = 0.003), and surgery performed in the second half of the day (OR, 0.016; 95% CI, 1.266-10.398; P = 0.016). Operating on phakic patients compared with pseudophakic or aphakic patients (OR, 0.962; 95% CI, 5.049-57.644; P < 0.001) and the use of endotamponade (OR, 5.288; 95% CI, 1.769-15.813; P = 0.003) were associated with a reduced risk for endophthalmitis. In culture-positive endophthalmitis, the presence of diabetes (OR, 4.61; 95% CI, 1.15-18.39; P = 0.03), vitreous substitute (BSS or RL) (OR, 6.08, 95% CI, 1.47-25.10, P = 0.012), and pseudophakia (OR, 5.68; 95% CI, 1.37-23.47; P = 0.016) were significant risk factors. Conclusions: Patients who are immunocompromised, pseudophakic, or aphakic are at a higher risk of endophthalmitis after PPV. Endotamponade significantly mitigates the risk of infection after vitrectomy surgery. (C) 2018 by the American Academy of Ophthalmology

Human conjunctival flora plays an important role in protecting the eye. Contact lens wearers are prone to have changes in conjunctiva flora. This study aims at assessing the variability of conjunctiva flora among contact lens wearers. We performed Ion Torrent based 16s metagenomic sequencing from 4 contact lens wearers conjunctiva swab samples. Proteobacteria found to be the most predominant phylum present in all samples. On genus level we have detected 250 bacterial strains and we found upto 455 species bacterial strains at class level, the sample was found to be most enriched with Alpha proteobacteria (52%), Gamma proteobacteria (18%) and Actinobacteria (9%) and Bacilli (6%). Order level, analysis showed presence of 52% of Alpha proteobacteria with Rhodobacterales (29%), Sphingomanadales (20%), and 1% Caulobacterales order followed by 18% of Gamma proteobacteria with Pseudomonadales (15%), Xanthomanadales (1%), 13% of Beta proteobacteria occupies Burkholderiales and 5% of Bacillales. Wearing contact lens may increase the risk of eye infections and also from the data we found that opportunistic infections may not only be caused by predominant species like cultivable Pseudomanas but also be due to uncultivable bacteria found in conjunctival swab samples the detection of which was made possible by next generation sequencing.

Purpose: To evaluate the effect of lignocaine on growth and apoptosis indication of primary human Tenon’s capsule fibroblast (HTFs) in an in vitro model. Patients and Methods: Tenon’s capsule tissue obtained from patients undergoing trabeculectomy were grown in cell culture medium. The effect of different concentrations of lignocaine (0.5, 1.0, 1.5, and 2%) on the morphology and growth of the fibroblasts was studied using microscopy, cell viability, and proliferation assay, and apoptosis was detected using the FITC Annexin V Apoptosis Kit. Results: Morphological changes similar to those of apoptotic cells, including cytoplasmic vacuolation, shrinkage, and rounding were visualized in the cells treated with concentrations greater than 1.0% (i.e., 1.5, 2.0%). Though proliferation inhibition was found with all four concentrations (0.5–2.0%), the viability of cells decreased from 1.0% lignocaine. Conclusion: 0.5% lignocaine prevents proliferation of fibroblasts without causing apoptosis in vitro.

Human adenoviruses (HAdVs) are uniquely important "model organisms" as they have been used to elucidate fundamental biological processes, are recognized as complex pathogens, and are used as remedies for human health. As pathogens, HAdVs may effect asymptomatic or mild and severe symptomatic disease upon their infection of respiratory, ocular, gastrointestinal, and genitourinary systems. High-resolution genomic data have enhanced the understanding of HAdV epidemiology, with recombination recognized as an important and major pathway in the molecular evolution and genesis of emergent HAdV pathogens. To support this view and to actualize an algorithm for identifying, characterizing, and typing novel HAdVs, we determined the DNA sequence of 95 isolates from archives containing historically important pathogens and collections housing currently circulating strains to be sequenced. Of the 85 samples that were completely sequenced, 18 novel recombinants within species HAdV-B and D were identified. Two HAdV-D genomes were found to contain novel penton base and fiber genes with significant divergence from known molecular types. In this data set, we found additional isolates of HAdV-D53 and HAdV-D58, two novel genotypes recognized recently using genomics. This supports the thesis that novel HAdV genotypes are not limited to "one-time" appearances of the prototype but are of importance in HAdV epidemiology. These data underscore the significance of lateral genomic transfer in HAdV evolution and reinforce the potential public health impact of novel genotypes of HAdVs emerging in the population.

Infectious keratitis is a major cause of blindness, next to cataract and majority of cases are mainly caused by gram negative bacterium Pseudomonas aeruginosa (P. aeruginosa). In this study, we investigated a P. aeruginosa VRFPA10 genome which exhibited susceptibility to commonly used drugs in vitro but the patient had poor prognosis due to its hyper virulent nature. Genomic analysis of VRFPA10 deciphered multiple virulence factors and P.aeruginosa Genomic Islands (PAGIs) VRFPA10 genome which correlated with hyper virulence nature of the organism. The genome sequence has been deposited in DDBJ/EMBL/GenBank under the accession numbers LFMZ01000001-LFMZ01000044.

Laryngeal carcinoma is the second commonest and most aggressive cancer originating in the squamous cells of the laryngeal epithelium. Undesirable complications and resistance to chemotherapy are the common problems which necessitate the development of alternative anticancer agents for the management of laryngocarcinoma. We herein report the biosynthesis of gold nanoparticles using Muntingia calabura (Mc-AuNps) and their anticancer efficacy against Hep2 cells line. The biologically synthesized gold nanoparticles were characterized by UV-visible spectroscopy, dynamic light scattering (DLS), Fourier transform infrared spectroscopy (FTIR) and transmission electron microscopy (TEM) study. The Mc-AuNps exhibited excellent monodispersity with an average diameter of 27 nm, with the negative zeta potential of about -18 mV as well as long-term stability. Our results showed that Mc-AuNps induced cytotoxicity in Hep2 cells and inhibited the cell proliferation by inducing apoptosis. Further study revealed that Mc-AuNps-induced apoptosis in Hep2 cells was associated with disruption of cell membrane, alteration of nuclear morphology and cell cycle arrest at G2 phase. These results clearly suggested that the green synthesis of Mc-AuNps could be used to treat laryngeal cancer cells.

Pseudomonas aeruginosa (P. aeruginosa) is a menacing opportunistic, nosocomial pathogen; become a growing concern as conventional antimicrobial therapy is now futile against it. Multi-drug resistant P. aeruginosa (MDRPA) has distinctive resistance mechanisms such as production of β-lactamases, repression of porin genes and over-expression of efflux pumps. The focus of this study is to standardize and application of multiplex PCR (mPCR) to detect the presence of betalactamase genes encoding blaTem, blaOXA, blaCTX-M-15, blaVim, blaGes, blaVeb, blaDIM, AmpC and Efflux pump genes encoding Mex A,B-oprM, Mex C,D-oprJ, Mex X,Y-oprN, oprD, nfxB, MexR. A total of 200 clinical isolates of P. aeruginosa were tested for the presence of the above mentioned genes genotypically through mPCR and characterized by phenotypic methods for ESBL and MBL production. Out of 200 isolates, 163 (81.5%) nfxB regulator gene, 102 (51%) MexA, 96 (48%) MexC, 93 (46.5%) MexB, 86 (43%) MexD, 81 (40.5%) OprM, 74 (37%) OprJ, 72 (36%) OprD and MexR, 53 (26.5%) Mex X and OprN, 49 (24.5%) MexY gene. Betalactamase genes 145 (72.5%) blaTem, 67 (33.5%) blaOXA, 35 (17.5%) blaVim, 25(12.50%), 23 (11.50%) blaVeb, 21 (11.5%) blaGes, 14 (7%) Ctx-m and 10 (5%) AmpC and 5 (2.5%) blaDim-1 gene were tested positive by mPCR. Phenotypically 38 (19%) and 29 (14.5%) out of 200 tested positive for ESBL and MBL production. Application of this mPCR on clinical specimens is fast, accurate, specific and low-cost reliable tool for the screening, where culture negative Eubacterial PCR positive cases for an early molecular detection of drug resistance mechanism assisting the clinician to treat the disease with appropriate antibiotic selection.

Although intravitreal injections are considered a great advancement in the management of retinal diseases because they have potential to improve vision, endophthalmitis is a concern to a treating ophthalmologist. 1 Dossarps D. Bron A.M. Koehrer P. et al. FRCR net (FRenCh Retina specialists net). Endophthalmitis after intravitreal injections: incidence, presentation, management, and visual outcome. Am J Ophthalmol. 2015; 160: 17-25 Abstract Full Text Full Text PDF PubMed Scopus (102) Google Scholar , 2 Mithal K. Mathai A. Pathengay A. et al. Endophthalmitis following intravitreal anti-VEGF injections in ambulatory surgical centre facility: incidence, management and outcome. Br J Ophthalmol. 2013; 97: 1609-1612 Crossref PubMed Scopus (17) Google Scholar Previous studies have found a prevalence of postinjection endophthalmitis of 0% to 0.87%. The purpose of this study is to report the incidence, microbiological profile, antimicrobial susceptibility, clinical findings, management, and visual outcomes in patients with acute postinjection endophthalmitis (within 6 weeks) after anti-vascular endothelial growth factor and intravitreal injection of triamcinolone acetonide (IVTA). Re: Raman et al.: Five-year incidence and visual outcomes in postintravitreal injection endophthalmitis (Ophthalmology 2016;123:1162-4)OphthalmologyVol. 123Issue 11PreviewWe read with great interest the article by Raman et al1 on the 5-year incidence and visual outcomes in postintravitreal injection endophthalmitis. We commend the authors for their comprehensive study.1 Full-Text PDF

We read with great interest the article by Raman et al1Raman R. Singh H.S. Appanraj R. et al.Five-year incidence and visual outcomes in postintravitreal injection endophthalmitis.Ophthalmology. 2016; 123: 1162-1164Abstract Full Text Full Text PDF PubMed Scopus (4) Google Scholar on the 5-year incidence and visual outcomes in postintravitreal injection endophthalmitis. We commend the authors for their comprehensive study.1Raman R. Singh H.S. Appanraj R. et al.Five-year incidence and visual outcomes in postintravitreal injection endophthalmitis.Ophthalmology. 2016; 123: 1162-1164Abstract Full Text Full Text PDF PubMed Scopus (4) Google Scholar Infections with Burkholderia species are difficult to treat owing to the multidrug resistance of the bacteria. Proper antibiotics must be chosen based on antibiotic susceptibility patterns of the isolated strains.2Ornek K. Ozdemir M. Ergin A. Burkholderia cepacia keratitis with endophthalmitis.J Med Microbiol. 2009; 58: 1517-1518Crossref PubMed Scopus (17) Google Scholar However, the details of antibiotic susceptibility of Burkholderia species isolated from bevacizumab vial were not provided. The authors found that the bevacizumab vial was contaminated with Burkholderia species and accounted that as the cause of the outbreak. However, the diagnosis of Burkholderia species endophthalmitis was not confirmed in the study. Because of the poor sensitivity of conventional microbiological tests like microscopy and culture,3Melo G.B. Bispo P.J. Yu M.C. et al.Microbial profile and antibiotic susceptibility of culture-positive bacterial endophthalmitis.Eye (Lond). 2011; 25: 382-387Crossref PubMed Scopus (47) Google Scholar it was not surprising that the clinical samples from cluster endophthalmitis patients tested for microscopy and cultures were negative. Although one of the cluster endophthalmitis samples was reported to be positive for eubacterial polymerase chain reaction, the polymerase chain reaction–amplified product was not sequenced to identify the bacteria.4Loh J. Lee J.Y. Kaur D.O.G.S.R. et al.Abiotropia defectiva-related endophthalmitis in an immunocompromised patient and application of 16s rRNA in microbiologic diagnosis.Ocul Immunol Inflamm. 2016; : 1-6Crossref Scopus (1) Google Scholar For the confirmation of the source and to prove that it was the cause of the outbreak, ideally DNA sequencing could have been applied on clinical sample and bacterial culture isolated from bevacizumab vial to study the clonal relationship between bacterial strains. It would also be interesting to know about the concomitant use of contaminated bevacizumab vial for any other patients and if any other vials of the same batch tested for microbial colonization. Details about the management of cluster endophthalmitis, including what antibiotics were used and if it was different from treatment of other sporadic cases of postintravitreal injection endophthalmitis cases, were not provided. Use of improperly fractionated bevacizumab is the major risk factor for postintravitreal injection endophthalmitis outbreaks. The United States Pharmacopeia formulated specific guidelines to prevent such catastrophic incidences. These guidelines highly recommend the use of a device called a minispike dispensing pin with security clips for preparing the compounding drugs.5Gonzalez S. Rosenfeld P.J. Stewart M.W. et al.Avastin doesn't blind people, people blind people.Am J Ophthalmol. 2012; 153: 196-203.e1Abstract Full Text Full Text PDF PubMed Scopus (76) Google Scholar We would, therefore, like to know if authors practice the use of such dispensing pins for fractionation and what were the other sterile precautions followed by the authors to fractionate the bevacizumab in their study. In addition, what further actions were taken to prevent similar outbreaks of cluster endophthalmitis and if this incidence was reported to the manufacturer? Five-Year Incidence and Visual Outcomes in Postintravitreal Injection EndophthalmitisOphthalmologyVol. 123Issue 5PreviewAlthough intravitreal injections are considered a great advancement in the management of retinal diseases because they have potential to improve vision, endophthalmitis is a concern to a treating ophthalmologist.1,2 Previous studies have found a prevalence of postinjection endophthalmitis of 0% to 0.87%. The purpose of this study is to report the incidence, microbiological profile, antimicrobial susceptibility, clinical findings, management, and visual outcomes in patients with acute postinjection endophthalmitis (within 6 weeks) after anti-vascular endothelial growth factor and intravitreal injection of triamcinolone acetonide (IVTA). Full-Text PDF ReplyOphthalmologyVol. 123Issue 11PreviewWe thank Agrawal et al for the critical appraisal of our study on the 5-year incidence and visual outcomes in postintravitreal injection endophthalmitis. Full-Text PDF