Measuring fecal metabolites of endogenous steroids using ESI-MS/MS spectra in Taiwanese pangolin, (order Pholidota, family Manidae, Genus: Manis): A non-invasive method for endangered species.

Pangolins are 'keystone species' driven towards extinction due to a lack of profound awareness and illegal trade. The drivers urge for immediate development in the understanding of demographics and reproductive dynamics of this species. In this study, we developed and validated a quantitative method to measure pangolin fecal extracts using the electrospray (ESI-MS/MS) interface in positive ionization mode. The method aids in the measurement of hormones from the hypothalamic-pituitary-adrenal (HPA) and hypothalamic-pituitary-gonadal (HPG) axis, making it a possibly appropriate technique to understand the cross-talk between the axes. The study aims to measure the relative abundance of adrenal and gonadal hormones such as corticosterone, cortisol, estrone, estradiol-17ß, progesterone, testosterone, and a number of its metabolites. From the dried fecal extract, the principal metabolite identified from the estrogen family was estradiol-17ß, whereas the gestagen family revealed that the pregnane series is predominated in 5α-configuration. On the other hand, epiandrosterone was seen as the dominant form in the male fecal extracts. Additionally, the glucocorticoids are excreted majorly as corticosterone, but traces of cortisol are also present in both the male and female fecal samples. The physiological validation confirmed that the ESI-MS/MS technique is suitable to determine physiologically caused differences in the fecal steroid concentrations. Physiologically, the age structure in pangolin is not responsible for causing differences within gender. However, the results revealed that glucocorticoids might vary between the sexes, i.e., males have a higher relative abundance of glucocorticoids over females. Therefore, our studies show that some of the main adrenal and gonadal metabolites can be predicted by exploiting MS/MS, which can steer research to potentially assess the reproductive status of captive and free-ranging pangolin species.

Women in surgery: trends in nine surgical specialties.

BACKGROUND: Gender disparity in surgery remains an issue worldwide. We analysed the trends in gender distribution in surgical subspecialties across Australia and New Zealand over the last 8 years. METHODS: Workforce reports from the Royal Australasian College of Surgeons were reviewed from 2014 to 2021. Data relating to 4802 applicants, 1554 trainees and 6839 active surgeons across nine surgical subspecialties was analysed. Statistical analysis was performed using a difference of proportions hypothesis test. Predictions regarding time to parity were performed using a linear regression model. RESULTS: All nine surgical subspecialties' surgeon numbers saw a steady increase in the representation of women between 2014 and 2021. The most significant rise seen in general surgery from 14% to 21% (P ≤ 0.001). Proportions of women trainees were variable, the only significant rise was seen in orthopaedics from 9% to 19% (P ≤ 0.001). Proportions of women applicants and successful applicants have also been variable. General surgery saw the only consistent increase in women applicants over the last 6 years, from 35% in 2016 to 43% in 2021. Linear regression predictions estimate that paediatric surgery will be the first to reach gender parity in 23 years, and orthopaedics the last, in 186 years. CONCLUSION: A steady increase in women has been observed across all surgical subspecialities over the last 8 years. However, estimates based on current trends suggest that gender parity may be out of reach for the next eight generations in subspecialties such as cardiothoracic and orthopaedic surgery.

Does histological prostatic inflammation during transurethral resection of the prostate for bladder outlet obstruction affect post-operative urinary outcomes?

OBJECTIVE: Benign prostate hyperplasia (BPH) is a common cause for bladder outlet obstruction (BOO) and lower urinary tract symptoms (LUTS) in men. The pathophysiology of BPH is multifactorial and inflammation has been linked with progression of BPH and LUTS. The association between histological prostatitis found at transurethral resection of the prostate (TURP) and adverse post-operative urinary outcomes is not clearly defined. Our aim was to evaluate the association between histological prostatitis and adverse post-operative urinary outcomes following TURP procedure. METHODS: Patients who had undergone TURP for BPH at a single institution between 2014 and 2018 were included. The study population was divided into three cohorts: those with no histological inflammation, those with any form of inflammation and those specifically with prostatic stromal inflammation. Functional outcomes were assessed by defining a series of measurable post-operative "LUTS events" and comparing these to time-to-event profile using a Kaplan-Meier estimator. RESULTS: A total 198 patients were included (no inflammation n = 101; any inflammation n = 97, prostatic stromal inflammation n = 81). All three groups were comparable in terms of baseline characteristics. The any inflammation group had significantly more adverse post-operative outcomes after TURP compared to the no inflammation group, P = 0.0065. The stromal inflammation group had more LUTS events after surgery compared to the no inflammation groups in the first year of follow-up n = 0.011; over a 5-year follow-up period the results were not statistically significant, P = 0.244. CONCLUSION: Histological prostatitis is associated with worse urinary outcomes after TURP compared to no inflammation. These results are useful in improving prognostic discussions with patients after TURP.

New horizons in the reproductive biology of Chinese pangolin (Manis pentadactyla) using the gonadal hormonal profile.

The reproductive uniqueness of pangolins has been documented through diverse biological reports with discernible data discrepancies in gestation, copulation, and pregnancy. These mechanistic reproductive differences have yet to be endocrinologically quantified, which could assist in optimizing natural breeding in zoos to recover endangered species. The present research characterizes the Chinese pangolin's annual seasonal reproductive pattern by measuring immunoreactive estrogens and progestagens in 34 captive females and testosterone in 29 captive males. Our results showed that Chinese pangolins are seasonal breeders, with most births witnessed during Sept-Dec, overlapping with the field records. Females exhibited spontaneous ovulation and post-partum ovulation. Pregnant females exhibited a higher P4 level for ~ 9 months (Jan-Sept) and decreased before parturition (Oct-Dec). The circulating E2 is maintained at the baseline in pregnant females year-round. Contrastingly, in non-pregnant females, P4 is maintained at the baseline, apart from a slight elevation in January, and E2 demonstrates a sudden hike from November and remains elevated until February, suggesting the onset of ovulation. The serum testosterone concentration in males peaked during October, which is in sync with the female ovulation period. As a result that their major reproductive events, ovulation, mating, and parturition, all transpire in November-March. Evidence also supports that Chinese pangolins exhibit signs of postimplantation (pregnancy) ranging only from 5 to 6 months (May-Oct), preceded by possible facultative delay implantation triggered by lactation. The provided data not only fill in the knowledge gap for this critically endangered species but can also assist in making informed decisions, which can directly affect the successful breeding of this species in captivity.

Teaching brief motivational interviewing to medical students using a pedagogical framework.

OBJECTIVE: Medical schools are charged with assisting medical students to acquire the confidence, knowledge and skills for behavior change conversations in primary healthcare. The present study evaluated teaching brief motivational interviewing (MI) to pre-clinical medical students. METHODS: Forty-six students participated in an educational intervention premised on the Learn, See, Practice, Prove, Do, Maintain pedagogical framework, comprising 2 × 2-h lectures, a 2-h role-play triad session, and 3 × 2-h small group simulated patient encounters supported by scaffolding strategies. Measures of brief MI knowledge (MI Knowledge and Attitudes Test & Multiple-Choice Knowledge Test) and confidence (MI Confidence Scale) were taken at baseline, post-training, and 3-month follow-up, and skills (Behavior Change Counseling Index) were assessed at three intervals during simulated patient encounters. RESULTS: Students who received brief MI training improved in knowledge and confidence from baseline to post-training and gains remained at 3-months. Brief MI skills improved across the simulation sessions. CONCLUSION: Pre-clinical medical students can attain knowledge, confidence and skills in brief MI after participation in a short intervention and improvements are sustainable. PRACTICE IMPLICATIONS: Our results support the use of an evidence-based pedagogical framework for teaching brief MI in pre-clinical years of medical curricula and our scaffolding strategy affords promise.

Bioinformatic and immunological investigation of Per a 5 (delta class GST) allergen from Periplaneta americana.

INTRODUCTION: GSTs are multifunctional enzymes involved in cellular detoxification and present as potent allergens in several sources. Present study investigates allergenic relevance of GST from P. americana and determine its cross reactive potential with other indoor allergen sources. METHODS: Computational analysis with FASTA and ConSurf webserver was performed to determine potentially cross reactive allergens. Further, Per a 5 gene was cloned in pET 22b+ vector and expressed in E.coli BL21 cells and the rPer a 5 protein was purified using Ni-NTA affinity chromatography. Enzymatic activity of rPer a 5 was assessed using CDNB and cumene hydroperoxide. ELISA and immunoblot were performed using cockroach hypersensitive patient's sera. Functional activity of rPer a 5 was evaluated by basophil activation test. Inhibition studies were carried out with D. pteronyssinus, A. alternata and C. lunata extracts. RESULTS: Per a 5 demonstrates highest sequence similarity with delta class GST of Blattella germanica (94.9%). It also exhibits significant sequence similarity (50-58%) with mite, fungal and helminth allergenic GSTs. ConSurf analysis reveals high degree of evolutionary similarity in N terminal region of Per a 5, especially at GST dimerization interface. The purified rPer a 5 protein resolved at 27 kDa on SDS-PAGE. The rPer a 5 protein exhibits GST activity and possess upto 65% immunoreactivity with cockroach hypersensitive patient's sera in ELISA and immunoblot. It upregulates expression of CD203c on basophils signifying its biological ability to activate effector cells. rPer a 5 significantly inhibits corresponding GSTs in P. americana, D. pteronyssinus, A. alternata and C. lunata with EC50 values of 15.5 ng. 38.38 ng, 41.4 ng and 61.66 ng, respectively. CONCLUSION: Recombinant delta class GST of P. americana is a clinically relevant allergen showing upto 65% immunoreactivity with hypersensitive patient's sera. Per a 5 GST allergen showed phylogenetic similarity with dust mite, fungal and birch allergens thereby demonstrating allergen cross reactivity.

Identification of a vicilin-like major allergen from Prosopis juliflora exhibiting cross- reactivity with legume food allergens.

BACKGROUND: Prosopis juliflora is a clinically relevant allergic sensitizer worldwide and shares cross-reactivity with allergens from several tree pollen and food. The present study aims to purify and immunobiochemically characterize a major allergen from Prosopis pollen. The allergen was further investigated for its cross-reactivity with legume allergens. METHODS: Prosopis extract was fractionated by Q Sepharose and Superdex 75 gel filtration column to purify the allergen. Specific IgE against purified protein was estimated via ELISA and immunoblot. The protein was subjected to mass spectrometric analysis. Glycan characterization was performed by Schiff staining and lectin binding assay followed by deglycosylation studies. The functional activity of the purified protein was evaluated by the basophil activation test. Cross-reactivity was assessed by inhibition studies with legume extracts. RESULTS: A 35 kDa protein was purified and showed 75% IgE reactivity with the patients' sera by ELISA and immunoblot. Glycan characterization of protein demonstrated the presence of terminal glucose and mannose residues. A reduction of 40% and 27% in IgE binding was observed upon chemical and enzymatic deglycosylation of the protein, respectively. The glycoprotein allergen upregulates the expression of CD203c on basophils which was significantly reduced upon deglycosylation, signifying its biological ability to activate the effector cells. The identified protein shared significant homology with Lup an 1 from the lupine bean. Immunoblot inhibition studies of the purified allergen with legume extracts underlined high cross-reactive potential. Complete inhibition was observed with peanut and common bean, while up to 70% inhibition was demonstrated with soy, black gram, chickpea, and lima bean. CONCLUSION: A 35 kDa vicilin-like major allergen was isolated from P. juliflora. The protein possesses glycan moieties crucial for IgE binding and basophil activation. Furthermore, the purified protein shows homology with Lup an 1 and exhibits cross-reactivity with common edible legume proteins.

Mortality and morbidity in wild Taiwanese pangolin (Manis pentadactyla pentadactyla).

Globally, pangolins are threatened by poaching and illegal trade. Taiwan presents a contrary situation, where the wild pangolin population has stabilized and even begun to increase in the last two decades. This paper illustrates the factors responsible for causing mortality and morbidity in the wild Taiwanese pangolin (Manis pentadactyla pentadactyla) based on radio-tracking data of wild pangolins and records of sick or injured pangolins admitted to a Taiwanese wildlife rehabilitation center. Despite being proficient burrowers, results from radio-tracking show that Taiwanese pangolins are highly susceptible to getting trapped in tree hollows or ground burrows. Data from Pingtung Rescue Center for Endangered Wild Animals showed that trauma (73.0%) was the major reason for morbidity in the Taiwanese pangolin with trauma from gin traps being the leading cause (77.8%), especially during the dry season, followed by tail injuries caused by dog attacks (20.4%). Despite these threats, Taiwan has had substantial success in rehabilitating and releasing injured pangolins, primarily due to the close collaboration of Taiwanese wildlife rehabilitation centers over the last twenty years.

Chemical Crosslinking: Role in Protein and Peptide Science.

BACKGROUND: Chemical crosslinking refers to intermolecular or intramolecular joining of two or more molecules by a covalent bond. The reagents that are used for the purpose are referred to as 'crosslinking reagents' or 'crosslinkers'. Based on factors like reactivity and spacer length these are classified into different types, each having its own specific function and application. In recent times, chemical crosslinking has emerged as an efficient tool for the study of biomolecules like proteins. It finds its application in various studies including the attachment of proteins to a solid support for the study of membrane receptors, protein-protein complexes, protein-DNA complexes, and others. When coupled with techniques like mass spectroscopy, it has been used not only for the determination of three dimensional structures of proteins but also for the study of protein-protein interactions and determination of interesting sites. This combination of mass spectrometry techniques and bioinformatics, added yet another dimension to our present day understanding of protein chemistry. Thus, chemical crosslinking has multitude uses that it can be put to. METHODS: We undertook a systematic search of bibliographic databases and search engine such as Google Scholar, Scifinder, Scopus, Mendeley etc for review of research literature. We excluded research paper which only reported synthesis of crosslinker molecules and did not involve any mass spectrometry studies. RESULTS: Sixty-four papers were included in the review. The majority of references were taken from last ten years as there has been an immense progress in this area in the recent years. Eleven classical papers in this field were included which talk about basic of this methodology. Thirty-two papers discussed about various types of organic groups used for designing chemical cross-linkers and various methodologies which were used to enhance the crosslinking efficiency. These papers also highlight various strategies used to enhance detection of cross-linked proteins and various computer software used to detect cross-linking sites from mass data. Twenty-one papers showed the proof concept application of this methodology to detect protein crosslinking in-vivo and in-vitro. CONCLUSION: The findings of this review confirm the importance chemical crosslinking combined with mass spectroscopy as a low cost alternative to understand protein-protein interaction. The information generated by this methodology can help in better understating of various diseases and for the development of better drugs for them.

Pulsatile Clavicular Swelling as the Sole Presentation of Asymptomatic Renal Cell Carcinoma with an Isolated Metastasis to the Clavicle: A Case Report.

Renal Cell Carcinoma (RCC) is notorious for its unpredictable dissemination patterns involving both lymphatic and haematogenous route without a clear-cut preponderance for any. Unlike other intra-abdominal malignancies like colorectal carcinoma, renal cell carcinoma is often known to metastasize to distant sites even after a confident and unsuspecting curative resection or often as the primary presentation constantly adding and agitating our presentation patterns for this sneaky tumour. Presented below is a case of such an unsuspecting female patient. She was referred from the orthopaedic OPD to surgery OPD with a right clavicular swelling which was histopathologically examined and diagnosed as a metastatic renal cell carcinoma following which she was diagnosed with right renal cell carcinoma.

Uso de nosódios na prática clínica: resultados de uma enquete / Use of nosodes in homeopathic clinical practice: a survey

Os nosódios são preparações homeopáticas de materiais orgânicos derivados de produtos inativos de doenças, culturas de microrganismos (bactérias, vírus e fungos, por exemplo) ou parasitas, material infectado ou com alterações patológicas ou produtos de decomposição animal ou humana feitos seguros através do procedimento farmacotécnico homeopático. Os nosódios têm papel importante na prática clínica. São frequentemente indicados como medicamentos usuais, intercorrentes, antimiasmáticos, agudos, etc., de acordo com a percepção individual de cada profissional. Assim, há variações no uso clínico desses medicamentos, que pode estar influenciado pelo conhecimento, experiência, proficiência e noções preconcebidas dos profissionais. A presente pesquisa objetivou investigar o uso real dos nosódios na prática clínica homeopática.

Nosodes are homeopathic preparations of organic materials derived from inactivated disease products, cultures of microorganisms (e.g., bacteria, fungi and viruses) or parasites, infected or pathologically changed material or decomposition products from humans or animals, rendered safe during the homeopathic manufacturing process. In homeopathic clinical practice, nosodes have an important and indispensable part. They are frequently indicated as common, intercurrent, anti-miasmatic, or acute remedies, etc., depending on the physician’s perception. But there are variations in their clinical use, which might be influenced by the background knowledge, experience, expertise and preconceived notions of practitioners. The present survey sought to find the actual use made of nosodes in homeopathic clinical practice.

Seroprevalence of HIV, HBV, HCV and syphilis in blood donors in Southern Haryana.

Blood transfusion is an important mode of transmission of infections to recipients. The aim of the study was to assess the prevalence of transfusion-transmissible infections among blood donors. For this, a 3.5-year retrospective study, from October 2002 to April 2006 was conducted at the blood transfusion centre of Maharaja Agrasen Medical College, Agroha (Hisar) Haryana. Donors were screened for seroprevalence of HIV, HBV, HCV and syphilis. A total of 5849 donors were tested, out of which 4010 (68.6%) were replacement donors and 1839 (31.4%) were voluntary donors. The seroprevalence of HIV was 0.3% in the donors. No voluntary donor was found to be positive for HIV. The low sero-positivity among donors is attributed to pre-donation counseling in donor selection. The seroprevalence of HBV, HCV and syphilis was 1.7%, 1.0% and 0.9% respectively in total donors. The seroprevalence of hepatitis and syphilis was more in replacement donors as compared to voluntary donors.

Human trichinosis in remotes of Uttarakhand, India.

Trichinosis is a parasitic zoonosis caused by ingestion of infected meat containing larvae of Trichinella, more prevalent in developing countries. Although infection with Trichinella is globally distributed, it has been documented only rarely in India. The reports are available where Trichinella larvae were found from animals in India but, to our knowledge, only one human case has been reported from India (Punjab), so far. This is the first report of small multiple outbreaks of human trichinosis in India (2009-2011). Epidemiological, clinical, and laboratory findings of trichinosis patients belonging to remote areas of Uttarakhand were analyzed retrospectively and prospectively. Patients belonged to remote areas of Garhwal, Uttarakhand, 77.78% were male, and 22.22% were female. The age of patients ranged from 9-55 yrs. History of eating meat of wild boar was given by all (100%). The signs and symptoms of the patients varied even after intake of same diet, and included generalized weakness/malaise, myalgia, fever, gastrointestinal symptoms, facial/periorbital edema, subconjunctival hemorrhages, retinal hemorrhages, muscle atrophy, and dyspnea. Laboratory investigations revealed eosinophilia, leukocytosis, creatine phosphokinase (CPK) and serum glutamic oxaloacetic transaminase (SGOT) elevation in 100%, 88.89%, 50% and 16.67%, respectively. Muscle biopsies revealed larvae in 27.78%. One patient expired while others improved. The prevalence of trichinosis is likely to be underestimated. The aim of this study is to emphasize on the magnitude of the problem, to educate people, especially in the affected areas about this health hazard and help implementation of epidemiological studies and preventive measures.