Production and partial purification of β-galactosidase enzyme from probiotic Bacillus subtilis SK09

139-144In the present study, lactose hydrolyzing enzyme β-galactosidase was produced from a novel probiotic Bacillus subtilis SK09 and it was compared with commercially obtained, B. subtilis MTCC 2413. The probiotic Bacilli was initially screened for its ability to hydrolyze the X-gal followed by production of blue colonies. The production medium was formulated using varied nutrients including carbon, nitrogen, mineral salts and amino acid sources. The crude enzyme with an activity of 76.2 U/ml was achieved from our novel strain B. subtilis SK09, and subjected for partial purification by solvent precipitation, dialysis and ion exchange chromatography using DEAE cellulose (DEAE-C) column. Partially purified β-galactosidase enzyme showed 11.6 folds increase in specific activity of 137.02 U/ml. This partially purified enzyme was further characterized and it was observed that it is homogeneous in nature, with molecular weight of 43 kDa. When compared, the β-galactosidase activity of our novel strain was found five times higher than that of commercially obtained strain (27.6 U/ml). Hence, this characteristic of B. subtilis SK09 of increase enzymatic activity can make it a promising candidate for various industrial as well as biotechnological applications

Identification of Patala (Stereospermum colais and Stereospermum suaveolens roots) by pharmacognostic parameters - A plant drug in Dasamula

Stereospermum colais and Stereospermum suaveolens are known as "Patala" in Ayurveda and also a constituent of Dasamula. It is difficult to distinguish between the two species and hence it is difficult to identify genuine Patala. Both Stereospermum species looks alike in morphology with the exception of flower-color. So collection of the plant material from wild source as well as differentiating from the marketed sample is a challenge for the herbal industries to identify the genuine Patala. Hence, an attempt was made to compare the pharmacognostical and phytochemical parameters of the roots of S. colais and S. suaveolens. Macroscopy, microscopy, physico-chemical analysis and elemental analysis were carried out to standardize the roots. The salient diagnostic features identified to distinguish the plant species are heterocellular periderm and calcium oxalate druses in S. colais and multitype (Rhytidome) periderm and calcium oxalate raphides in S. suaveolens. Qualitative and quantitative phytochemical analysis and comparative HPTLC fingerprint analysis of various extracts of roots revealed their phytochemical composition. The standardization parameters developed here can be used as reference standard for correct identification of the plant. Further, it will act as a tool to detect adulterants and substituents consequently maintains the quality, reproducibility and efficacy of the plant material.

Cortical Gyrification in Velo-Cardio-Facial (22q11.2 Deletion) Syndrome: A Longitudinal Study

Introduction: Velo-cardio-facial syndrome (VCFS) has been identified as an important risk factor for psychoses, with up to 32% of individuals with VCFS developing a psychotic illness. Individuals with VCFS thus form a unique group to identify and explore early symptoms and biological correlates of psychosis. In this study, we examined if cortical gyrification pattern, i.e. gyrification index (GI) can be a potential neurobiological marker for psychosis. Method: GIs of 91 individuals with VCFS were compared with 29 siblings and 54 controls. Further, 58 participants with VCFS, 21 siblings and 18 normal controls were followed up after 3 years and longitudinal changes in GI were compared. Additionally, we also correlated longitudinal changes in GI in individuals with VCFS with prodromal symptoms of psychosis on the Scale of Prodromal Symptoms (SOPS). Result: Individuals with VCFS had significantly lower GIs as compared to their siblings and normal controls. Longitudinal examination of GI did not reveal any significant group–time interactions between the three groups. Further, longitudinal change in GI scores in the VCFS group was negatively correlated with positive prodromal symptoms, with the left occipital region reaching statistical significance. Conclusion: The study confirms previous reports that individuals with VCFS have reduced cortical folding as compared to normal controls. However over a period of three years, there is no difference in the rate of change of GI among both individuals with VCFS and normal controls. Finally, our results suggest that neuroanatomical alterations in areas underlying visual processing may be an early marker for psychosis

First-in-Human Study of MANP: A Novel ANP (Atrial Natriuretic Peptide) Analog in Human Hypertension

[Figure: see text]

MANP Activation Of The cGMP Inhibits Aldosterone Via PDE2 And CYP11B2 In H295R Cells And In Mice

Background: Aldosterone is a critical pathological driver for cardiac and renal diseases. We recently discovered that mutant atrial natriuretic peptide (MANP), a novel atrial natriuretic peptide (ANP) analog, possessed more potent aldosterone inhibitory action than ANP in vivo. MANP and natriuretic peptide (NP)-augmenting therapy sacubitril/valsartan are under investigations for human hypertension treatment. Understanding the elusive mechanism of aldosterone inhibition by NPs remains to be a priority. Conflicting results were reported on the roles of the pGC-A (particulate guanylyl cyclase A receptor) and NP clearance receptor in aldosterone inhibition. Furthermore, the function of PKG (protein kinase G) and PDEs (phosphodiesterases) on aldosterone regulation are not clear. Methods: In the present study, we investigated the molecular mechanism of aldosterone regulation in a human adrenocortical cell line H295R and in mice. Results: We first provided evidence to show that pGC-A, not NP clearance receptor, mediates aldosterone inhibition. Next, we confirmed that MANP inhibits aldosterone via PDE2 (phosphodiesterase 2) not PKG, with specific agonists, antagonists, siRNA silencing, and fluorescence resonance energy transfer experiments. Further, the inhibitory effect is mediated by a reduction of intracellular Ca2+ levels. We then illustrated that MANP directly reduces aldosterone synthase CYP11B2 (cytochrome p450 family 11 subfamily b member 2) expression via PDE2. Last, in PDE2 knockout mice, consistent with in vitro findings, embryonic adrenal CYP11B2 is markedly increased. Conclusions: Our results innovatively explore and expand the NP/pGC-A/3',5', cyclic guanosine monophosphate (cGMP)/PDE2 pathway for aldosterone inhibition by MANP in vitro and in vivo. In addition, our data also support the development of MANP as a novel ANP analog drug for aldosterone excess treatment

EFFECT OF GEMCITABINE ON FEMALE FERTILTY – AN ANIMAL MODEL STUDY

Background and objectives: Advances in cancer chemotherapy have led to improved survival rates and poses greater emphasis on preserving quality of life post-treatment. Gonadotoxicity is a well-recognized side effect of many cancer chemotherapeutic agents. This study was designed to evaluate the effects of gemcitabine, an antimetabolite anticancer drug, on oogenesis in Swiss albino mice and its reversibility. Methods: Thirty six inbred female Swiss albino mice in diestrous phase were selected and divided into three groups of twelve each. Groups were labelled as A, B and Control. Groups A and B received 40 mg/kg and 80 mg/kg of gemcitabine intraperitoneally. The control group received saline intraperitoneally. At the end of two weeks 6 mice were sacrificed from each group and the rest at the end of 2 months.  Ovaries were studied histologically. Results: After 2 weeks, the ovaries of experimental group mice showed more number of atretic follicles as compared to control group (p<0.01). The diameter of corpus lutea was more, though a reduction in number was recorded in experimental group (p<0.05).  Whereas after 2 months, both the experimental groups showed no difference in terms of atretic follicles, diameter and number of corpus lutea (p>0.05). Conclusions: These findings suggest that administration of gemcitabine may have profound effects on oogenesis and hence female fertility. This study also suggests that the effects are reversible. Keywords: Gemcitabine; Oogenesis; Swiss albino mice; atretic follicles; corpus luteum

The Rsr1/Bud1 GTPase Interacts with Itself and the Cdc42 GTPase during Bud-Site Selection and Polarity Establishment in Budding Yeast

Bimolecular fluorescence complementation assays allow the visualization of the homotypic and heterotypic GTPase interactions in vivo. The Rsr1 homotypic interaction involves its polybasic region and depends on its GDP-GTP exchange factor. Dimerization of GTPases may be an efficient mechanism to set up cellular asymmetry

Mortality and pulmonary complications in patients undergoing surgery with perioperative SARS-CoV-2 infection: an international cohort study

Background: The impact of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) on postoperative recovery needs to be understood to inform clinical decision making during and after the COVID-19 pandemic. This study reports 30-day mortality and pulmonary complication rates in patients with perioperative SARS-CoV-2 infection. Methods: This international, multicentre, cohort study at 235 hospitals in 24 countries included all patients undergoing surgery who had SARS-CoV-2 infection confirmed within 7 days before or 30 days after surgery. The primary outcome measure was 30-day postoperative mortality and was assessed in all enrolled patients. The main secondary outcome measure was pulmonary complications, defined as pneumonia, acute respiratory distress syndrome, or unexpected postoperative ventilation. Findings: This analysis includes 1128 patients who had surgery between Jan 1 and March 31, 2020, of whom 835 (74·0%) had emergency surgery and 280 (24·8%) had elective surgery. SARS-CoV-2 infection was confirmed preoperatively in 294 (26·1%) patients. 30-day mortality was 23·8% (268 of 1128). Pulmonary complications occurred in 577 (51·2%) of 1128 patients; 30-day mortality in these patients was 38·0% (219 of 577), accounting for 81·7% (219 of 268) of all deaths. In adjusted analyses, 30-day mortality was associated with male sex (odds ratio 1·75 [95% CI 1·28–2·40], p\textless0·0001), age 70 years or older versus younger than 70 years (2·30 [1·65–3·22], p\textless0·0001), American Society of Anesthesiologists grades 3–5 versus grades 1–2 (2·35 [1·57–3·53], p\textless0·0001), malignant versus benign or obstetric diagnosis (1·55 [1·01–2·39], p=0·046), emergency versus elective surgery (1·67 [1·06–2·63], p=0·026), and major versus minor surgery (1·52 [1·01–2·31], p=0·047). Interpretation: Postoperative pulmonary complications occur in half of patients with perioperative SARS-CoV-2 infection and are associated with high mortality. Thresholds for surgery during the COVID-19 pandemic should be higher than during normal practice, particularly in men aged 70 years and older. Consideration should be given for postponing non-urgent procedures and promoting non-operative treatment to delay or avoid the need for surgery. Funding: National Institute for Health Research (NIHR), Association of Coloproctology of Great Britain and Ireland, Bowel and Cancer Research, Bowel Disease Research Foundation, Association of Upper Gastrointestinal Surgeons, British Association of Surgical Oncology, British Gynaecological Cancer Society, European Society of Coloproctology, NIHR Academy, Sarcoma UK, Vascular Society for Great Britain and Ireland, and Yorkshire Cancer Research