Phase variable colony variants are conserved across Gardnerella spp. and exhibit different virulence-associated phenotypes.

The Gardnerella genus, comprising at least 13 species, is associated with the polymicrobial disorder bacterial vaginosis (BV). However, the details of BV pathogenesis are poorly defined, and the contributions made by individual species, including Gardnerella spp., are largely unknown. We report here that colony phenotypes characterized by size (large and small) and opacity (opaque and translucent) are phase variable and are conserved among all tested Gardnerella strains, representing at least 10 different species. With the hypothesis that these different variants could be an important missing piece to the enigma of how BV develops in vivo, we characterized their phenotypic, proteomic, and genomic differences. Beyond increased colony size, large colony variants showed reduced vaginolysin secretion and faster growth rate relative to small colony variants. The ability to inhibit the growth of Neisseria gonorrhoeae and commensal Lactobacillus species varied by strain and, in some instances, differed between variants. Proteomics analyses indicated that 127-173 proteins were differentially expressed between variants. Proteins with increased expression in large variants of both strains were associated with amino acid and protein synthesis and protein folding, whereas those increased in small variants were related to nucleotide synthesis, phosphate transport, ABC transport, and glycogen breakdown. Furthermore, whole genome sequencing analyses revealed an abundance of genes associated with variable homopolymer tracts, implicating slipped strand mispairing in Gardnerella phase variation and illuminating the potential for previously unrecognized heterogeneity within clonal populations. Collectively, these results suggest that phase variants may be primed to serve different roles in BV pathogenesis.IMPORTANCEBacterial vaginosis is the most common gynecological disorder in women of childbearing age. Gardnerella species are crucial to the development of this dysbiosis, but the mechanisms involved in the infection are not understood. We discovered that Gardnerella species vary between two different forms, reflected in bacterial colony size. A slow-growing form makes large amounts of the toxin vaginolysin and is better able to survive in human cervix tissue. A fast-growing form is likely the one that proliferates to high numbers just prior to symptom onset and forms the biofilm that serves as a scaffold for multiple BV-associated anaerobic bacteria. Identification of the proteins that vary between different forms of the bacteria as well as those that vary randomly provides insight into the factors important for Gardnerella infection and immune avoidance.

Association of periodontal disease with oral lichen planus: A systematic review and meta analysis.

Background: Gingival lesions in oral lichen planus can indirectly increase the risk of plaque-induced periodontal disease when symptoms associated with such lesions hamper the proper oral hygiene maintenance by the patients and can increase the risk of periodontal tissue destruction. This systematic review analyses the existing evidence on the association between oral lichen planus and periodontal disease. Aim: This systematic review of case-control studies aimed to analyse the association between periodontal disease and oral lichen planus. Material and Methods: An electronic database search for randomised controlled trials, experimental studies, case-control studies, and cohort studies published in peer-reviewed Journals in the English language was conducted from the following databases: PubMed, EBSCOHost, Science Open, EMBASE, and Google Scholar. Results: A total of 12,507 were identified on an electronic database search. Only eight studies fulfilled the eligibility criteria and were included for quantitative analysis. A data extraction sheet was prepared, and studies were analysed. Conclusion: Bleeding on Probing and Probing depth were seen to be significantly associated with Oral Lichen Planus. The symptoms in Oral Lichen Planus impede efficient oral hygiene maintenance by a patient and predispose them to the occurrence of long-term Periodontal Disease.

Endogenous fungal endophthalmitis following intensive corticosteroid therapy in severe COVID-19 disease.

Purpose: To report endogenous fungal endophthalmitis, postrecovery from severe COVID-19 infection in otherwise immunocompetent individuals, treated with prolonged systemic steroids. Methods: Retrospective chart review of cases with confirmed and presumed fungal endogenous endophthalmitis, following severe COVID-19 disease, treated at two tertiary care referral eye institutes in North India. Results: Seven eyes of five cases of endogenous fungal endophthalmitis were studied. All cases had been hospitalized for severe COVID-19 pneumonia and had received systemic steroid therapy for an average duration of 42 ± 25.1 days (range 18-80 days). All the cases initially complained of floaters with blurred vision after an average of 6 days (range 1-14 days) following discharge from hospital. They had all been misdiagnosed as noninfectious uveitis by their primary ophthalmologists. All eyes underwent pars plana vitrectomy (PPV) with intravitreal antifungal therapy. Five of the seven eyes grew fungus as the causative organism (Candida sp. in four eyes, Aspergillus sp. in one eye). Postoperatively, all eyes showed control of the infection with a marked reduction in vitreous exudates and improvement in vision. Conclusion: Floaters and blurred vision developed in patients after they recovered from severe COVID-19 infection. They had received prolonged corticosteroid treatment for COVID-19 as well as for suspected noninfectious uveitis. We diagnosed and treated them for endogenous fungal endophthalmitis. All eyes showed anatomical and functional improvement after PPV with antifungal therapy. It is important for ophthalmologists and physicians to be aware of this as prompt treatment could control the infection and salvage vision.

Lacosamide reduces HDAC levels in the brain and improves memory: Potential for treatment of Alzheimer's disease.

Lacosamide, a histone deacetylase (HDAC) inhibitor, has been approved for the treatment of epilepsy. Some HDAC inhibitors have been proven effective for the treatment of memory disorders. The present investigation was designed to evaluate the effect of lacosamide on memory and brain HDAC levels. The effect on memory was evaluated in animals with scopolamine-induced amnesia using the elevated plus maze, object recognition test, and radial arm maze. The levels of acetylcholinesterase and HDAC in the cerebral cortex were evaluated. Lacosamide at doses of 10 and 30mg/kg significantly reduced the transfer latency in the elevated plus maze. Lacosamide at a dose of 30mg/kg significantly increased the time spent with a familiar object in the object recognition test at the 24h interval and decreased the time spent in the baited arm. Moreover, at this dose, the number of errors in the radial arm maze at 3 and 24h intervals was minimized and a reduction in the level of HDAC1, but not acetylcholinesterase, was observed in the cerebral cortex. These effects of lacosamide are equivalent to those of piracetam at a dose of 300mg/kg. These results suggest that lacosamide at a 30mg/kg dose improves disrupted memory, possibly by inhibiting HDAC, and could be used to treat amnesic symptoms of Alzheimer's disease.

Tumor mechanics and metabolic dysfunction.

Desmosplasia is a characteristic of most solid tumors and leads to fibrosis through abnormal extracellular matrix (ECM) deposition, remodeling, and posttranslational modifications. The resulting stiff tumor stroma not only compromises vascular integrity to induce hypoxia and impede drug delivery, but also promotes aggressiveness by potentiating the activity of key growth, invasion, and survival pathways. Intriguingly, many of the protumorigenic signaling pathways that are mechanically activated by ECM stiffness also promote glucose uptake and aerobic glycolysis, and an altered metabolism is a recognized hallmark of cancer. Indeed, emerging evidence suggests that metabolic alterations and an abnormal ECM may cooperatively drive cancer cell aggression and treatment resistance. Accordingly, improved methods to monitor tissue mechanics and metabolism promise to improve diagnostics and treatments to ameliorate ECM stiffening and elevated mechanosignaling may improve patient outcome. Here we discuss the interplay between ECM mechanics and metabolism in tumor biology and suggest that monitoring these processes and targeting their regulatory pathways may improve diagnostics, therapy, and the prevention of malignant transformation.

Role of taxanes in the treatment of advanced NHL patients: A randomized study of 87 cases.

UNLABELLED: NHL is a highly chemo-sensitive as well as radiosensitive disease. From May 2005 to June 2010, 87 patients were randomised into 2 arms. The control arm received the standard CHOP regimen + IFRT, whereas the study arm received Paclitaxel, 135/m2 additionally. The results showed a better Overall Response (87% vs 78%) in the study arm. The 3yr and 5yr overall survival were significantly better in the study arm (89% vs 77%, p- value <0.05; 83% vs 67%, p-value <0.05). However, the incidence and severity of the side effects, haematological and non- haematological were enhanced but manageable in the study arm. KEYWORDS: Taxanes, Paclitaxel, Non Hodgkin's Lymphoma.

Regulation of Cdk7 activity through a phosphatidylinositol (3)-kinase/PKC-ι-mediated signaling cascade in glioblastoma.

The objective of this research was to study the potential function of protein kinase C (PKC)-ι in cell cycle progression and proliferation in glioblastoma. PKC-ι is highly overexpressed in human glioma and benign and malignant meningioma; however, little is understood about its role in regulating cell proliferation of glioblastoma. Several upstream molecular aberrations and/or loss of PTEN have been implicated to constitutively activate the phosphatidylinositol (PI) (3)-kinase pathway. PKC-ι is a targeted mediator in the PI (3)-kinase signal transduction repertoire. Results showed that PKC-ι was highly activated and overexpressed in glioma cells. PKC-ι directly associated and phosphorylated Cdk7 at T170 in a cell cycle-dependent manner, phosphorylating its downstream target, cdk2 at T160. Cdk2 has a major role in inducing G(1)-S phase progression of cells. Purified PKC-ι phosphorylated both endogenous and exogenous Cdk7. PKC-ι downregulation reduced Cdk7 and cdk2 phosphorylation following PI (3)-kinase inhibition, phosphotidylinositol-dependent kinase 1 knockdown as well as PKC-ι silencing (by siRNA treatment). It also diminished cdk2 activity. PKC-ι knockdown inhibited overall proliferation rates and induced apoptosis in glioma cells. These findings suggest that glioma cells may be proliferating through a novel PI (3)-kinase-/PKC-ι/Cdk7/cdk2-mediated pathway.

Development and validation of a liquid chromatography/tandem mass spectrometry assay for the simultaneous determination of nateglinide, cilostazol and its active metabolite 3,4-dehydro-cilostazol in Wistar rat plasma and its application to pharmacokinetic study.

Nateglinide (NTG), an insulin secretogogue, has been studied in rats for drug-drug interaction with cilostazol (CLZ), an antiplatelet agent commonly used in diabetics. We developed a liquid chromatography tandem mass spectrometry (LC-MS/MS) based method that is capable of simultaneous monitoring plasma levels of nateglinide, cilostazol, and its active metabolite 3,4-dehydro-cilostazol (DCLZ). All analytes including the internal standard (Repaglinide) were chromatographed on reverse phase C(18) column (50 mm x 4.6mm i.d., 5 microm) using acetonitrile: 2mM ammonium acetate buffer, pH 3.4 (90:10, v/v) as mobile phase at a flow rate 0.4 ml/min in an isocratic mode. The detection of analyte was performed on LC-MS/MS system in the multiple reaction monitoring (MRM) mode. The quantitations for analytes were based on relative concentration. The method was validated over the concentration range of 20-2000 ng/ml and the lower limit of quantitation was 20 ng/ml. The recoveries from spiked control samples were >79% for all analytes and internal standard. Intra- and inter-day accuracy and precision of validated method were with in the acceptable limits of <15% at all concentration. The quantitation method was successfully applied for simultaneous estimation of NTG, CLZ and DCLZ in a pharmacokinetic drug-drug interaction study in Wistar rats.

Endoscopic CSF rhinorrhea closure: our experience in 267 cases.

OBJECTIVES: The advances in endoscopic sinus surgery have made it the procedure of choice for treatment of cerebrospinal fluid rhinorrhea (CSFR). To analyze the efficacy of endoscopic closure of CSFR was the objective of the present study. METHODS: We treated 267 patients with CSFR endoscopically. Diagnosis was achieved with the help of CT with or without cisternography, MRI, beta-2 transferrin levels. Fascia lata and fat were used to plug the defects. The patients were followed up for a minimum period of 6 months. RESULTS: CSFR was successfully plugged in 258 patients. Nine patients required revision surgery. S ix could be plugged successfully endoscopically and 2 patients by a neurosurgical approach. CONCLUSIONS: In our experience with 267 patients of endoscopically treated CSFR, the results of achieving the closure was 96.63% in the first instance and 98.88% after revision surgery. SIGNIFICANCE: The transnasal endoscopic approach has excellent results in the treatment of CSFR. We recommend it as the optimum surgical approach for both primary and revisional surgical management of CSFR.

Expression of GSTM1 in angiofibromas.

The aetiology of juvenile nasopharyngeal angiofibroma (JNA) is largely unknown. In this study, we have investigated the expression of glutathione S transferase M1 (GSTM1) gene in angiofibroma. The GSTM1 allele gene locus is normally found in all human beings. When this is not expressed there is an increased risk of developing a malignancy of the upper aerodigestive tract. In this study, we have assessed eight samples of JNA for the expression of GSTM1, by the polymerase chain reaction (PCR) followed by polyacrylamide gel electrophoresis (PAGE), three of the eight patients studied failed to express this gene. Further investigation in this area is warranted.