The normative values for resting length of pectoralis minor muscle among individuals with asymptomatic shoulder in South Bengaluru: a cross-sectional pilot study.

OBJECTIVE: The aim of the study was to determine the normative values for resting length of pectoralis minor muscle among males and females with an asymptomatic shoulder in South Bengaluru. PATIENTS AND METHODS: Two hundred and forty-six subjects with asymptomatic shoulders were taken by convenience sampling. The subjects were divided into two groups: Group A (123 males) and Group B (123 females). Pectoralis minor muscle resting length was measured on their dominant side in all subjects. Post measurement, the PMI was calculated. The normative values for both groups were determined. The mean PMI was compared between Group A and Group B and was analyzed using statistical tools. RESULTS: In Group A, the mean average Pectoralis minor length (PML) was 14.59 ± 1.61 cm and in Group B, the mean average PML was 12.95 ± 1.42 cm which was statistically significant (p-value <0.00001). In Group A, the mean Pectoralis Minor Index (PMI) was 8.54 ± 0.88 and in Group B, the mean PMI was 8.22 ± 0.90 which was statistically significant (p-value <0.005). CONCLUSIONS: The normative values for resting length of pectoralis minor muscle for males are 8.54 ± 0.88 and for females 8.22 ± 0.90 with an asymptomatic shoulder. There is a difference in the normative values for the resting PML in the asymptomatic shoulder by gender.

Avidin-Biotin recombinant nucleoprotein competitive ELISA for the detection of peste des petits ruminants virus antibodies in sheep and goats.

The present study describes the development of a truncated recombinant peste des petits ruminants virus (PPRV) nucleoprotein (rPPRV-NPN) and its polyclonal antibodies-based immuno-diagnostic assay, Avidin-Biotin (AB) recombinant nucleoprotein competitive ELISA (ABrC-ELISA) for the detection of PPRV antibodies in the sheep and goats. The PPRV N-terminal immunogenic region (1-266 aa) of nucleoprotein (NPN) coding sequence was amplified and cloned into the pETite vector. The rPPRV-NPN with a molecular weight of ∼ 30 kDa was expressed in E. coli, purified, and characterized by SDS-PAGE and immunoblot using standard PPRV specific sera. The Ni-NTA affinity-purified rPPRV-NPN as coating antigen and its hyperimmune serum as competitive antibodies raised in guinea pigs were evaluated as diagnostic reagents in ABrC-ELISA using the known standard panel of sera. The threshold (cut-off) Percentage Inhibition (PI) value was determined as 45 (mean ± 3 SD) based on the reactivity of the known sheep and goats sera to PPRV antibodies [negative (n = 140) and positive (n = 98)] and the assay had a sensitivity of 97 % (95 % Confidence Interval (CI): 91.3-99.4 %) and specificity of 100 % (95 % CI: 97.4-100 %) with an excellent Area under curve (AUC) of 0.997 (95 % CI: 0.99-1.0). On evaluation of diagnostic performance of the assay using the sheep and goats sera (n = 391) from vaccinated, infected, and non-vaccinated animals, the ABrC-ELISA showed the relative diagnostic sensitivity of 95.88 % (95 % CI: 92.56-98.01 %) & 98.77 % (95 % CI: 96.43-99.74 %) and diagnostic specificity of 97.97 % (95 % CI: 94.19-99.58 %) & 90.54 % (95 % CI: 84.64-94.73 %) against indigenous PPR competitive ELISA kit & IDvet Screen® PPR Competition kit, respectively. The study showed that ABrC-ELISA is rapid, sensitive, and specific and can be a better alternative assay for the detection of the PPRV antibodies in the sera of small ruminants for serosurveillance / seromonitoring of PPR not only at the eradication and post-eradication phases in the disease-controlled endemic countries but also in the PPR non-endemic countries.

Evaluation of recombinant leptospiral surface antigen (Lsa27) lipoprotein for serodiagnosis of human leptospirosis by latex agglutination test.

PURPOSE: Leptospirosis has wide clinical presentations often mimicking other illnesses, thus rapid and simple diagnostics will have facilitated the initial patient management and therapy compared to other inaccessible and laborious tests/assays. METHOD: In this study, the sensitized latex beads coated with purified recombinant outer membrane (OM)-leptospiral surface antigen (Lsa27) lipoprotein of pathogenic Leptospira was evaluated as a diagnostic antigen in latex agglutination test (LAT) for the detection of anti-leptospiral antibodies in the human sera. The prepared rLsa27 latex beads were evaluated with the confirmed microscopic agglutination test (MAT) reactive (at 1:50) Leptospira-specific positive (n = 42) and non-reactive negative (n = 80) sera from human cases suspected of leptospirosis with the history of pyrexia of unknown origin. RESULT: The results revealed the relative diagnostic sensitivity of 90.48 % (confidence interval (CI) at 95 % : 77.4-97.3 %) and diagnostic specificity of 91.35 % (CI at 95 %: 82.8-96.4 %), with an accuracy of 90.98 % (CI at 95 %: 84.44-95.41 %), and the kappa value of 0.8036 ±â€¯0.056 SE (CI at 95 %: 0.69-0.91) with a substantial agreement against gold standard serological MAT. CONCLUSION: The findings suggest that the rLsa27 protein-based LAT can be useful as a simple rapid screening diagnostic test for the detection of anti-leptospiral antibodies in the sera of humans. This rapid test can be complemented by other confirmatory diagnostics for the early detection of Leptospira antibodies which may in turn help in the prompt treatment and mitigates the public health problem at primary health care level.

Evaluation of the diagnostic potential of recombinant leptospiral OMP A-like protein (Loa22) and transmembrane (OmpL37) protein in latex agglutination test for serodiagnosis of leptospirosis in animals.

Leptospirosis is a re-emerging zoonotic disease of animals and humans caused by pathogenic Leptospira, which has major public health concerns. The study is aimed to express the recombinant outer membrane protein (OMP) A-like protein (rLoa22) and transmembrane (rOmpL37) protein of Leptospira interrogans serovar Hardjo in the Escherichia coli and their evaluation as a diagnostic antigen in the latex agglutination test (LAT) to detect anti-leptospiral antibodies in the sera of animals. The Loa22 and OmpL37 genes lacking signal peptide coding sequences were individually amplified (522 and 963 bp), by polymerase chain reaction, and directionally cloned into a pETite N-His Kan vector for expression. The expressed purified proteins were characterized by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and immunoblot, which confirmed leptospiral specific reactive protein with a molecular weight of ~19 and 36 kDa, respectively. The sensitized latex beads coated with these OM proteins separately were evaluated in LAT using cattle sera of microscopic agglutination test (MAT) confirmed positive (n = 53) and negative (n = 52) cases of leptospirosis. The rLoa22 LAT and rOmpL37 LAT revealed the relative diagnostic sensitivity of 94·34 and 96·23%, diagnostic specificity of 92·31 and 96·15% and accuracy of 93·33 and 96·19%, with the excellent agreement of Cohen's kappa value of 0·87 and 0·92, respectively. After extensive evaluation, this rapid recombinant protein-based field diagnostic test can be applied as a screening test for the detection of anti-leptospiral antibodies in the sera of animals in the field conditions.

Avidin-Biotin recombinant antigen capture ELISA for the detection of peste des petits ruminants virus in the clinical specimens of sheep and goats.

This study describes the development of Avidin-Biotin recombinant Antigen Capture ELISA (ABrAC ELISA) for the detection of the peste des petits ruminants virus (PPRV) antigens in the clinical specimens of sheep and goats. The assay uses the truncated recombinant PPRV N-terminal immunogenic region of nucleoprotein (rPPRV-NPN) as a reference positive antigen and its polyclonal antibodies as capture/detective antibodies and the rabbit PPRV polyclonal antibodies as coating antibodies. The cut-off value was determined as double times the mean reactivity of blank control based on the reactivity of the PPR confirmed negative and positive control panel samples. On assessing the specificity with the related differential diagnosis of the disease-causing viruses and bacteria, the assay showed specific detective reactivity to PPRV. Further, on evaluation using clinical specimens (n-274) of sheep and goats, the assay showed that the relative diagnostic sensitivity of 86.49 % (95 % confidence interval (CI): 71.23-95.46 %) and diagnostic specificity of 96.20 % (95 % CI: 92.91-98.25 %) against PPRV nucleoprotein-specific monoclonal antibody-based sandwich-ELISA (PPR s-ELISA) kit, with an accuracy of 94.89 % (95 % CI: 91.58-97.18 %) and Cohen's Kappa value of 0.791 + 0.055 SE (95 % CI: 0.68-0.90) with substantial agreements. The ABrAC-ELISA is an alternative method of an immunoassay for the rapid, sensitive, and specific detection of the PPRV antigens m the clinical specimens of sheep and goats for surveillance or diagnosis of PPR. This study also shows that the rPPRV-NPN and its specific polyclonal antibodies could be the sustainable source of safe diagnostic reagents without the need to handle the infectious virus during the eradication and post-eradication phases in endemic countries like India or PPR non-endemic countries.

Seroprevalence of Peste des petits ruminants in small ruminants in the North Eastern Region of India.

A seroprevalence study of the peste des petits ruminants (PPR) in small ruminants was carried out in the different states (Assam, Manipur, Meghalaya, Mizoram, Nagaland, and Tripura) in the North Eastern Region (NER) of India using serum samples collected from April 2017 to March 2018. A total number of 4,163 sera [sheep (n = 508) and goats (n = 3,655)] collected from 345 epi­units/villages covering 176 municipalities in NER were screened by competitive ELISA kit for the detection of PPR virus antibodies. The results revealed that the seroprevalence of PPR in small ruminants in Assam, Manipur, Meghalaya, Mizoram, Nagaland, and Tripura was 34.3%, 10.3%, 4.7%, 15.7%, 14.7%, and 5.5%, respectively with an overall 14.5% prevalence.Association between the presence of antibodies and goats has been showed to be significant (p < 0.01) at the NER level level and within every single state. This manuscript highlights the need for continuous monitoring of this important disease as for the severe economic impact PPR may have in the affected countries.

Robotic-assisted renal transplantation with total extraperitonealization of the graft: experience of 34 cases.

The aim of the study is to elucidate the method of complete extraperitonealization of the graft while performing transperitoneal robotic renal transplant recipient operation. This is a retrospective study of 34 robotic-assisted kidney transplant (RAKT) utilizing our technique from July 2015 to June 2017. The study is performed in a quaternary private hospital setting. The surgery was performed using our novel peritoneal flap technique allowing complete extraperitonealization of the kidney. Total of 34 patients underwent RAKT in our hospital. Mean age was 40.6 ± 8.1 years, there were 25 males and 9 females. 30 had single vessel and four had double vessels. 27 patients received living donor graft while seven had deceased donor. The mean blood loss was 88 ± 51 mL, mean anastomotic time was 32 ± 3.3 min, mean total operative time was 145 ± 15 min, mean hospital stay was 5.8 ± 2.3 days, time to nadir creatinine was 4.3 ± 1.9 days, median creatinine level at the end of 6 months was 1.3 mg/dL. There were three open conversions in our series, one of which had delayed graft function requiring hemodialytic support. Total extraperitonealization of the graft reproduces closely the technique of open kidney transplant with good graft function. This would be a step toward the ultimate goal of performing a complete extraperitoneal robotic transplant. Further refinements in robotic instruments would make this a reality in near future.

Molecular characterization of a proteolytic bacterium in Panchagavya: An organic fertilizer mixture.

Fermented product of combination of five major substances obtained from cow, viz., urine, milk, ghee, curd, and dung, is known as Panchagavya. Its pro-agricultural and medicinal value has been traditionally known to Indian farmers from Vedic period. In this study, the proteolytic properties of Panchagavya were investigated using Skim Milk Agar (SMA) form, a commercially available Panchagavya product. Proteolytic bacteria, SNCK-3, was successfully isolated. Further identification using 16s rDNA sequencing revealed that SNCK-3 belonged to Acinetobacter spp., which is a species of biofertilizer group. This observation justified the pro-agricultural role of Panchagavya. The present study represents primary data and it is essential to develop a new area of research for exposing the invisible or dormant Vedic biotechnological concepts, like Panchagavya.

Overexpression of heat shock GroEL stress protein in leptospiral biofilm.

Leptospira is the causative agent of leptospirosis, which is an emerging zoonotic disease. Recent studies on Leptospira have demonstrated biofilm formation on abiotic surfaces. The protein expressed in the biofilm was investigated by using SDS-PAGE and immunoblotting in combination with MALDI-TOF mass spectrometry. The proteins expressed in Leptospira biofilm and planktonic cells was analyzed and compared. Among these proteins, one (60 kDa) was found to overexpress in biofilm as compared to the planktonic cells. MALDI-TOF analysis identified this protein as stress and heat shock chaperone GroEL. Our findings demonstrate that GroEL is associated with Leptospira biofilm. GroEL is conserved, highly immunogenic and a prominent stress response protein in pathogenic Leptospira spp., which may have clinical relevance.

Molecular detection of pathogenic leptospiral protein encoding gene (lipL32) in environmental aquatic biofilms.

UNLABELLED: Leptospirosis is a zoonotic disease often encountered during post-monsoon season due to exposure with contaminated water. Leptospires have long been regarded as solitary organisms that persist in soil and aquatic environments. Here, the presence of leptospires in the aquatic biofilm exposed in the paddy field, sewers and stagnant rain water was demonstrated. Biofilm samples from paddy field water, submerged paddy leaves, sewers and stagnant rain waters from urban and rural areas were collected. Total genomic DNA was extracted and pathogenic leptospiral specific gene amplification was carried out to determine the spatial distribution of the bacteria. The degree of pathogenic Leptospira in biofilms from paddy field surface water, submerged leaf, were 33·3% and 27·2% respectively, whereas in rural and urban area, the sampling sites such as stagnant rain water, domestic sewer and collective sewers showed 11·1%, 13% and 16·6% with leptospires respectively. Higher proportion of pathogenic Leptospira in aquatic ecosystems, such as paddy field, could be one of the main factors for the occurrence of disease, more among the agricultural workers. This study would help to identify various survival strategies of leptospires in the environment and thus disease transmission. SIGNIFICANCE AND IMPACT OF THE STUDY: Little is known regarding the mechanisms by which pathogenic leptospires persist in aqueous environment, outside the mammalian host. In this view this is the first report of the distribution of Leptospira in environmental biofilm such as sewers and paddy leaf surfaces. This ability of pathogenic Leptospira to survive in aquatic ecosystems especially in biofilms could be one of the main factors which facilitate its survival in the environment, and thus disease transmission among the risk groups, such as sewage and agriculture worker. This study will encourage researchers in the field to consider biofilm as an important factor, when detecting leptospires in environment.

GC/MS profiling, in vitro anti-leptospiral and haemolytic activities of Boesenbergia rotunda (L.) Mansf. used as a medicinal plant by Nicobarese of Andaman and Nicobar Islands.

Leaves of the plant Boesenbergia rotunda are used by the Nicobarese tribe of Andaman and Nicobar Islands, India, to prepare traditional medicine for treating fever, headache and body ache. In the present investigation, methanol fraction of these leaves were analysed by GC/MS that revealed the presence of 25 compounds. The anti-leptospiral activity of methanol crude extract was determined by both microdilution and macrodilution methods. The MICs of the extract were tested against 24 pathogenic leptospiral strains and ranged between 62.5-125 µg/mL in both microdilution and macrodilution. The range of MBCs was 250 and 500 µg/mL in macrodilution and microdilution respectively. The crude extract was subjected to cytotoxic studies and found to have negligible or no haemolytic activity, exhibiting IC50 values of greater than 4 mg/mL. Further in vivo studies are needed to investigate the pharmacological and toxicological properties of Boesenbergia rotunda, before it can be considered as a new anti-leptospiral agent.

Use of Tetra-ammonium Tetrakis(4-Sulphonato)Phenyl Porphyrin for Pseudomonas and Bacillus Cell Imaging.

The use of tetraammonium tetrakis(4-sulphonato)phenyl porphyrin (TPPS), a water-soluble anionic compound, as a stain to analyse bacterial cells using fluorescent microscopy was investigated. TPPS was effectively used to analyse two different bacteria: Pseudomonas aeruginosa and Bacillus cereus. The variation in brightness with varying concentrations of TPPS was studied. The patterns of variations for these bacteria were found to be the same, but with consistently higher brightness for Bacillus cereus.