Effect of 4-methoxy 1-methyl 2-oxopyridine 3-carbamide on Staphylococcus aureus by inhibiting UDP-MurNAc-pentapeptide, peptidyl deformylase and uridine monophosphate kinase.

AIMS: The present study aimed to investigate the anti-Staphylococcus aureus and anti-biofilm properties of 4-methoxy-1-methyl-2-oxopyridine-3-carbamide (MMOXC) on S. aureus UDP-MurNAc-pentapeptide (MurF), peptidyl deformylase (PDF) and uridine monophosphate kinase (UMPK). METHODS AND RESULTS: The in vitro efficacy of MMOXC was evaluated using quantitative polymerase chain reaction, in vitro assays and broth microdilution methods. Further, the minimum inhibitory concentration (MIC), IC50 and zone of inhibition were recorded in addition to the anti-biofilm property. MMOXC inhibited pure recombinant UMPK and PDF enzymes with a Ki of 0·37 and 0·49 µmol l-1 . However Ki was altered for MurF with varying substrates. The MurF Ki for UMT, d-Ala-d-Ala and ATP as substrates was 0·3, 0·25 and 1·4 µmol l-1 , respectively. Real-time PCR analysis showed a significant reduction in PDF and MurF expression which correlated with the MIC90 at 100 µmol l-1 and IC50 in the range 42 ± 1·5 to 50 ± 1 µmol l-1 against all strains tested. At 5 µmol l-1 MMOXC was able completely to remove preformed biofilms of S. aureus and other drug resistant strains. CONCLUSIONS: MMOXC was able to kill S. aureus and drug resistant strains tested by inhibiting MurF, UMPK and PDF enzymes and completely obliterated preformed biofilms. SIGNIFICANCE AND IMPACT OF THE STUDY: Growth reduction and biofilm removal are prerequisites for controlling S. aureus infections. In this study MMOXC exhibited prominent anti-S. aureus and anti-biofilm properties by blocking cell wall formation, RNA biosynthesis and protein maturation.

Phenotyping at hot spots and tagging of QTLs conferring spot blotch resistance in bread wheat.

Spot blotch is a major foliar disease of wheat caused by Bipolaris sorokiniana in warm and humid environments of the world including South Asian countries. In India, it has a larger impact in Indo-Gangetic plains of the country. Therefore, the present study was undertaken to phenotype a mapping population at different hot spots of India and to detect quantitative trait loci (QTL) for resistance to spot blotch in wheat. For this study, 209 single seed descent (SSD) derived F8, F9, F10 recombinant inbred lines (RILs) of the cross 'Sonalika' (an Indian susceptible cultivar)/'BH 1146' (a Brazilian resistant cultivar) were assessed for spot blotch resistance at two hot spot locations (Coochbehar and Kalyani) for three years and for two years under controlled conditions in the polyhouse (Karnal). The population showed large variation in spot blotch reaction for disease severity in all the environments indicating polygenic nature of the disease. Microsatellite markers were used to create the linkage maps. Joint and/or individual year analysis by composite interval mapping (CIM) and likelihood of odds ratio (LOD) >2.1, detected two consistent QTLs mapped on chromosome 7BL and 7DL and these explained phenotypic variation of 11.4 percent and 9.5 percent over the years and locations, respectively. The resistance at these loci was contributed by the parent 'BH 1146' and shown to be independent of plant height and earliness. Besides, association of some agro-morphological traits has also been observed with percent disease severity. These identified genomic regions may be used in future wheat breeding programs through marker assisted selection for developing spot blotch resistant cultivars.

Molecular characterization of Orientia tsutsugamushi serotypes causing scrub typhus outbreak in southern region of Andhra Pradesh, India.

BACKGROUND & OBJECTIVES: Scrub typhus is a vector-borne zoonotic infection caused by Orientiatsutsugamushi. Local epidemiology of the circulating serotypes of scrub typhus is not available from most parts of India. We conducted this study for the diagnosis of scrub typhus using IgM ELISA and to detect O. tsutsugamushi serotypes circulating in southern Andhra Pradesh, India. METHODS: Samples were collected from patients clinically suspected to have scrub typhus and were subjected to IgM ELISA to measure IgM antibodies against O. tsutsugamushi. Nested polymerase chain reaction (PCR) was performed targeting strain-specific regions in ELISA-positive samples. RESULTS: Of a total of 663 samples, 258 (38.91%) were found to be positive by IgM ELISA. Serotypes could be detected in 230 (34.69%) samples only. Only two serotypes, Karp and Kawasaki, were found in the serum samples, with the former being predominant. The dual infection of Karp and Kawasaki serotypes was found in seven patients. Other serotypes such as Gilliam, Kuroki and Kato were not detected in the samples. INTERPRETATION & CONCLUSION: The nested PCR products proved useful in presumptively identifying the endemic O. tsutsugamushi serotypes. The present study could be significant in understanding scrub typhus epidemiology in this region.

A phase I trial of LHC165 single agent and in combination with spartalizumab in patients with advanced solid malignancies.

BACKGROUND: LHC165 is a Toll-like receptor (TLR)-7 agonist that generates an effective tumor antigen-specific T-cell adaptive immune response as well as durable antitumor responses. We aimed to evaluate the safety, tolerability, efficacy, dose-limiting toxicities, and pharmacokinetics (PK) of LHC165 single agent (SA) ± spartalizumab [PDR001; anti-programmed cell death protein 1 (PD-1)] in adult patients with advanced solid tumors. MATERIALS AND METHODS: In this phase I/Ib, open-label, dose-escalation/expansion study, patients received LHC165 SA 100-600 µg biweekly through intratumoral (IT) injection and LHC165 600 µg biweekly + spartalizumab 400 mg Q4W through intravenous (IV) infusion. RESULTS: Forty-five patients were enrolled: 21 patients received LHC165 SA, and 24 patients received LHC165 + spartalizumab. The median duration of exposure was 8 weeks (range 2-129 weeks). No maximum tolerated dose was reached. Recommended dose expansion was established as LHC165 600 µg biweekly as SA and in combination with spartalizumab 400 mg Q4W. The most common drug-related adverse events (AEs) were pyrexia (22.2%), pruritus (13.3%), chills (11.1%), and asthenia (4.4%). The only serious AE (SAE) suspected to be related to the study drug was grade 3 pancreatitis (n = 1). Across all tumor types, overall response rate and disease control were 6.7% and 17.8%, respectively. Overall median progression-free survival (PFS) and immune-related PFS was 1.7 months. LHC165 serum PK demonstrated an initial rapid release followed by a slower release due to continued release of LHC165 from the injection site. CONCLUSIONS: LHC165 demonstrated acceptable safety and tolerability both as SA and in combination with spartalizumab, and evidence of limited antitumor activity was seen in adult patients with relapsed/refractory or metastatic solid tumors.

Opportunistic invasive fungal pathogen Macrophomina phaseolina prognosis from immunocompromised humans to potential mitogenic RBL with an exceptional and novel antitumor and cytotoxic effect.

With the ever-increasing risk for fungal infections, one can no longer ignore fungi. It is imperative that clinical manifestations "presume fungus" with their epidemiologic and pathogenic features when evaluating a potentially infected patient. In the high-risk patient groups, fungi with intrinsic resistance to antifungal agents already exist, with a tendency to emerge as opportunistic pathogens. One of the smart pathogens is Macrophomina phaseolina, with the potential to disarm plant, animal, and human immunity. The response prophylaxis may vary from antifungal therapy and surgical measures to biochemical (Rhizoctonia bataticola lectin [RBL] with antitumor and cytotoxic nature) and gene therapeutics.

Isolation of Legionella pneumophila from clinical & environmental sources in a tertiary care hospital.

BACKGROUND & OBJECTIVES: Legionella pneumophila and other members of this genus are important respiratory pathogens but legionellosis often remains a neglected and under reported condition. Hence, this study was done to find out the presence of this organism in patients admitted to a tertiary care hospital with community-acquired pneumonia (CAP). METHODS: A total of 470 lower respiratory tract samples and 24 water samples from hospital outlets were examined. Culture was done on buffered charcoal yeast extract agar with supplements and identification of the isolates was done by microscopy and biochemical tests. RESULTS: L. pneumophila could be isolated from 12 (2.55%) patients suffering from community-acquired lower respiratory tract infection, unassociated with other aetiological agents of bacterial pneumonia. Of the 24 water samples tested, 8 (33.3%) grew the same organism. INTERPRETATION & CONCLUSION: Our study has shown that Legionella is present in the hospital environment and was the aetiological agent of lower respiratory tract infection in 2.55 per cent of patients. A larger study and reports from other parts of the country may help in determining the true significance of legionellosis in India.

An unusual case of obstructive jaundice.

Tuberculosis of the pancreas is a rarity, reported in a handful of literature. We enumerate the case of a young girl with high fever, jaundice, and right hypochondrial pain, whose investigations revealed a mass at the head of the pancreas. FNAC from the mass astoundingly proclaimed tuberculosis.

Aggressive papillary glioneuronal tumor: case report and literature review.

Papillary glioneuronal tumors (PGNT) are a rare, recently described form of mixed neoplasm composed of glial and neuronal components. PGNT usually occur in children and young adults, and typically demonstrate low-grade pathology, with a low proliferative index of 1-3%. Here we describe a newly diagnosed case of PGNT with a more aggressive phenotype that required irradiation and chemotherapy. The patient was a 19-year-old female who developed progressive headaches and visual seizures. An MRI revealed a heterogeneously enhancing solid mass in the left temporo-occipital region, with significant surrounding edema and mass effect. The mass was resected under stealth guidance without complication. Postoperative MRI scans showed patchy enhancement and residual T2 and FLAIR abnormality. Pathology revealed a highly cellular neoplasm with papillary-like structures, containing cells with glial and neuronal differentiation. Regions of mitoses and focal necrosis were noted, along with a Ki-67 labeling index of 26%. The diagnosis was aggressive PGNT, and treatment consisted of conformal irradiation and concomitant temozolomide over 6 weeks. Postirradiation follow-up MRI scans demonstrated a reduction of residual enhancement and FLAIR abnormality. The patient continues standard-dose adjuvant temozolomide on a monthly basis, with further improvement on subsequent MRI scans and a stable neurologic exam. This patient demonstrates that PGNT may, in rare cases, display an aggressive clinicopathologic phenotype that requires a therapeutic approach more consistent with a high-grade glioma.

Polycomb group genes control pattern formation in plant seed.

Transcriptional activators of the Trithorax group (TRX-G) and repressors of the Polycomb group (Pc-G) are involved in multiple aspects of embryogenesis in Drosophila and the mouse [1, 2] and appear to have a conserved role in the zygotic control of the development of the anterior-posterior axis [3, 4, 5]. In the model plant Arabidopsis, three Pc-G genes have been isolated and characterized to date. CURLY LEAF (CLF) represses the expression of a floral homeotic gene in vegetative tissues but does not appear to have a role in plant embryogenesis [6]. Two other Pc-G genes, FIS1/MEDEA [7, 8, 9], and FIS3/FIE [8, 10] have been characterized in studies of mutants that produce seeds in the absence of fertilization. Seeds resulting from autonomous development in fis mutants do not contain an embryo but only endosperm, the second product of double fertilization in flowering plants [11, 12]. Thus, FIS genes are considered to be repressors of endosperm development before fertilization. We report that when fis ovules are fertilized, the endosperm patterning along the major polar axis is perturbed. Posterior structures develop in more anterior domains of the endosperm. This correlates with the ectopic expression of a posterior molecular marker. FIS genes appear to be potent regulators of the establishment of the anterior-posterior polar axis in the endosperm.

Cerebral cortical lesions in multiple sclerosis detected by MR imaging at 8 Tesla.

BACKGROUND AND PURPOSE: Conventional imaging of ex-vivo brain at 1.5T in multiple sclerosis (MS) detects only a small fraction of the gray matter cerebral cortical lesions that can be detected by pathology. Our purpose was to examine if imaging at 8T can detect plaques in cortical gray matter (CGM) not evident at 1.5T. METHODS: An ex-vivo brain obtained at autopsy from a patient with MS was formalin fixed and 1 cm coronal slices were examined using MR imaging at 8T. RESULTS: Numerous cerebral cortical lesions not evident at 1.5T were seen at 8T. Lesions were easily identified using gradient-echo and spin-echo (SE) as well as diffusion images. MR imaging at 8T identified many of the types of plaques previously evident only by pathology. The magnitude of the cortical involvement in this 1 patient was severe. Lesions in the gray matter readily visible by high-field MR imaging were sometimes barely visible by pathology. MR imaging at 8T often facilitated the detection of such plaques by pathology. CONCLUSION: This study establishes the utility of high-field imaging at 8T in the delineation of plaques in the cerebral CGM in MS.

Ovule and embryo development, apomixis and fertilization.

Genetic analyses, particularly in Arabidopsis, have led to the identification of mutants that define different steps of ovule ontogeny, pollen stigma interaction, pollen tube growth, and fertilization. Isolation of the genes defined by these mutations promises to lead to a molecular understanding of these processes. Mutants have also been obtained in which processes that are normally triggered by fertilization, such as endosperm formation and initiation of seed development, occur without fertilization. These mutants may illuminate apomixis, a process of seed development without fertilization extant in many plants.

Comparative evaluation of recombinant HSP70 (N & C-terminal) fragments in the detection of equine trypanosomosis.

Trypanosomosis (Surra) is an economically important disease caused by Trypanosoma evansi which is an extracellular parasite present in the plasma, tissues and other body fluids of a wide range of hosts including domesticated animals. Currently, serological reports are based on detection of antibodies by ELISA using whole cell lysate (WCL) antigen, which has a limitation of persistence of anti-trypanosomal antibodies after successful treatment of the disease. Moreover, it has some ethical issues also like requirement of mice for in vivo maintenance of parasite for preparing the antigen. Therefore, in the present study, an attempt was made to evaluate the in vitro production of recombinant heat shock protein 70 (HSP70) for detection of antibodies in experimentally infected ponies. The amino acid sequence analysis of HSP70 revealed that N-terminal region of the protein was highly conserved while the C-terminal region was most divergent. The four different regions of HSP70 protein viz. HSP-1, HSP-2, HSP-3 and HSP-4 were cloned and expressed, among which HSP-1 (N-terminal region) & HSP-2 (C-terminal region) were truncated while HSP-3 & HSP-4 were complete C-terminal proteins. The recombinant fragments were probed with sequentially pooled experimental serum samples where antibodies were detected in these fragments from 10(th) day post infection till the termination of the experiment. Further, these recombinant fragments were also comparatively evaluated with WCL antigen in ELISA using experimental as well as field serum samples. It was observed that after successful treatment of infected ponies, there was a sharp fall in antibodies (within 90 days) when tested with recombinant HSP's fragments, while antibodies persisted even after 469 days when tested against WCL antigen. The sensitivity and specificity of all HSP70 fragments were also estimated from field serum samples with reference to WCL antigen ELISA. The HSP-1 showed minimum sensitivity (41.03%) among all the recombinant fragments. Among the C-terminal fragments, maximum sensitivity was observed with the HSP-2 (61.54%) while minimum was observed with HSP-4 (48.72%). The specificity increases for recombinant fragments from N-terminal to C-terminal region of protein and maximum specificity was observed with HSP-4 fragment (91.3%).

Lupus podocytopathy: An important differential diagnosis of nephrotic syndrome in systemic lupus erythematosus.

Some patients with systemic lupus erythematosus (SLE) present with sudden onset of nephrotic syndrome and biopsy findings may be of minimal change disease or focal segmental glomerulosclerosis with diffuse foot process effacement on electron microscopy but without significant immune deposits. This entity is termed lupus podocytopathy. Clinicians and renal pathologists need to be aware of this condition. Though steroid sensitive, it needs follow-up to recognize flare and class change, thereby optimizing therapy.

Seroprevalence of Scrub typhus at a tertiary care hospital in Andhra Pradesh.

INTRODUCTION: Scrub typhus is a rickettsial infection which is caused by Orientia tsutsugamushi and transmitted by the bite of the chigger of a mite. Delay in diagnosis can be fatal otherwise the treatment is simple, doxycycline being the drug of choice. Indirect immunoflurescence is considered gold standard but it is not used in India as it is costly and also not available. There is need for rapid, economic and simple test for the diagnosis of scrub typhus. This study was taken up to study the seroprevalence of scrub typhus in Andhra Pradesh and to compare two commonly used serological methods; rapid test and IgM ELISA. MATERIALS AND METHODS: This is a prospective study in which 100 serum samples from clinically suspected cases collected over a period of 3 months were processed for the detection of IgM antibodies for scrub typhus by ELISA and Rapid test. Samples were also tested for leptospirosis and dengue fever which the other common causes of fever prevalent in this region. RESULTS: Total number of samples processed was 100 of which 52 were males and 48 females. Among the hundred samples 39 were seropositive. Positivity was higher in the age group of patients between 16 and 30 yrs of age. There was 97% correlation between ELISA and rapid method. Of the 100 samples only three samples positive by ELISA were negative by rapid method. Fever was the most common manifestation and there was no eschar and no mortality reported. CONCLUSION: Scrub typhus should be included in the differential diagnosis of fever of unknown origin along with dengue, malaria and leptospirosis which are the other common endemic infections in this part of the country.

Cutaneous fungal infection in a renal transplantation patient due to a rare fungus belonging to order Pleosporales.

Fungal infections are being increasingly reported from immuno-compromised as well as immuno-competent patients. Transplant patients are on long term immunosuppressive therapy which makes them highly vulnerable to opportunistic fungal infections .These infections can be cutaneous or systemic. Several fungi have been reported to be the culprits such as Candida spp., Aspergillus spp., C. neoformans, P. carinii, and zygomycetes group of fungi. Cutaneous infections are most commonly caused by Pityriasis (tinea) versicolor, dermatophytes, and candida sp but these days the demtiaceous fungi are becoming more frequently reported .Here we report a case of post renal transplant cutaneous infection caused by dematiaceous fungus belonging to the order Pleosporales.

Bulimia nervosa in pregnancy: a case report.

Bulimia nervosa is an eating disorder characterized by secretive binge eating and purging with induced vomiting, laxatives, and diuretics. The disorder primarily afflicts young white women between 18-35 years of age. We report the case of a 30-year-old pregnant woman with a 17-year history of bulimia that involved up to six episodes of binging and purging daily. A multidisciplinary approach enabled outpatient management throughout gestation. The pregnancy resulted in the delivery of a normal 3000-g female infant at term. The ramifications for maternal and fetal well-being and the goals of therapeutic management are discussed.

Effect of bacterial growth on the bursting pressure of fetal membranes in vitro.

By mounting a layer of chorioamniotic membrane on a specially designed reaction vessel, we studied the effect of Escherichia coli and/or group B streptococcus growing on the decidual surface of the membranes in tissue culture or bacteriologic medium. The organisms grew equally well in either medium. When growing in tissue culture medium, either organism significantly weakened the membranes as compared with controls (membranes incubated in the absence of either organism). Membranes derived from pregnancies delivered vaginally or abdominally responded similarly. When organisms were grown in bacteriologic medium, bursting pressures did not decrease. Addition of bacteriologic medium (20-60%) to tissue culture medium did not affect bacterial growth, but inhibited significantly the lowering of bursting pressures. Bacteriologic medium also inhibited the peroxidase-H2O2-halide system in vitro. Heat-killed bacteria and/or supernatants of culture medium previously inoculated with bacteria were not effective in weakening membranes. The results suggest that live bacteria in conjunction with active membrane metabolism lead to a weakening and eventual rupture of the membranes.