A PRELIMINARY INVESTIGATION OF THE SHODHANA (DETOXIFICATION) OF ROOTS OF PLUMBAGO INDICA L. IN AYURVEDA

Plumbago indica L. (Plumbaginaceae) is a medicinal herb, credited with a number of therapeutic properties. It is widely used in Sri Lankan traditional medicine and in Ayurveda. In Sri Lanka, Ayurveda formulations which incorporate the roots of P. indica L. are prepared using commercially available air dried material after subjecting it to a detoxification with limewater prepared from commercially available milk of lime. The detoxification process is referred to as “Shodhanaâ€. According to the Ayurveda, this process is done to remove toxicity associated with the roots and, it can be surmised that it is done to remove toxicity associated with plumbagin, the predominant toxic naphthoquinone in P. indica L.. Shodhana of roots results in a deep maroon coloured extract arising from the calcium salt of plumbagin. Here, we report a qualitative and quantitative study of the Shodhana of roots of P. indica L. using Ultra Violet-Visible spectrophotometric and chromatographic methods to give a scientific basis for this process. A method for the quantitative extraction of plumbagin from root samples was optimized. A calibration curve for plumbagin in hexane was developed using the absorption values at 258 nm. The plumbagin content of 8.7 ± 0.1 mg/g in a fresh root sample was lowered by 19.4% to 7.0 ± 0.1 mg/g upon subjecting to Shodhana. It was found that a commercial dried root sample of P. indica L. root contained a much lower level (0.55 ± 0.05 mg/g) of plumbagin. To better understand the changes in the level of plumbagin in roots of P. indica L. during drying, a study was carried out by drying the root samples for five weeks subjecting them to analysis periodically by TLC and Ultra Violet-Visible spectrophotometric methods. The amount of plumbagin (8.6 ± 0.1 mg/g) present in fresh untreated roots at the beginning of the study was reduced by 62.7% to 3.2 ± 0.1 mg/g after one week. There was no measurable change in the plumbagin level thereafter up to week 5. The amount of plumbagin in dried roots can be further reduced by Shodhana. The total reduction of plumbagin by drying and Shodhana was 87.7%. Our results show that Shodhana of the fresh undried roots does not reduce the plumbagin content substantially, and that air drying followed by Shodhana is the most effective method to reduce the plumbagin content to a non-toxic level, supporting the currently used processing method

Feasibility and acceptability of a primary care liver fibrosis testing pathway centred on the diabetes annual review: PRELUDE1 prospective cohort study protocol.

INTRODUCTION: Non-alcoholic fatty liver disease is the most common chronic liver disease worldwide affecting 20%-25% in the USA and Europe with a 60%-80% lifetime prevalence for people with type 2 diabetes (T2D). Fibrosis has repeatedly been demonstrated to be the major determinant of liver disease morbidity and mortality and there is currently no routine screening for liver fibrosis in at-risk T2D population. METHODS AND ANALYSIS: This 12-month prospective cohort study of automated fibrosis testing uses the fibrosis-4 score (FIB-4) in patients with T2D linked to the investigation of hospital-based versus community-based second-tier transient elastography (TE) testing. We plan to include >5000 participants across 10 General Practitioner (GP) practices in East London and Bristol. This will determine the rate of undiagnosed significant liver fibrosis in a T2D population, the feasibility of two-tier liver fibrosis screening using FIB-4 at the diabetes annual review and subsequent TE delivered either in the community or secondary care settings. This will include an intention-to-treat analysis for all those invited to attend for diabetes annual review. A qualitative substudy regarding the acceptability of the fibrosis screening pathway will comprise semistructured interviews/focus groups with primary care staff (GPs and practice nurses), and patients taking part in the wider study. ETHICS AND DISSEMINATION: This study received a favourable opinion from the Cambridge East research ethics committee. The results of this study will be disseminated in peer-reviewed scientific journals, conference presentations and local diabetes lay panel meetings. TRIAL REGISTRATION NUMBER: ISRCTN14585543

Maximal Associated Regression: A nonlinear extension to Least Angle Regression

This paper proposes Maximal Associated Regression (MAR), a novel algorithm that performs forward stage-wise regression by applying nonlinear transformations to fit predictor covariates. For each predictor, MAR selects between a linear or additive fit as determined by the dataset. The proposed algorithm is an adaptation of Least Angle Regression (LARS) and retains its efficiency in building sparse models. Constrained penalized splines are used to generate smooth nonlinear transformations for the additive fits. A monotonically constrained extension of MAR (MARm) is also introduced in this paper to fit isotonic regression problems. The proposed algorithms are validated on both synthetic and real datasets. The performances of MAR and MARm are compared against LARS, Generalized Linear Models (GLM), and Generalized Additive Models (GAM) under the Gaussian assumption with a unity link function. Results indicate that MAR-type algorithms achieve a superior subset selection accuracy, generating sparser models that generalize well to new data. MAR is also able to generate models for sample deficient datasets. Thus, MAR is proposed as a valuable tool for subset selection and data exploration, especially when a priori knowledge of the dataset is unavailable

A comparison of Pentecostals in Hong Kong, Singapore and Kuala Lumpur: Culture and Belief

Using empirical and quantitative methods Pentecostal ministers are compared in the three locations of Hong Kong, Singapore and Kuala Lumpur. After providing an account of the historical backgrounds of Pentecostal churches in these locations, similarities and differences in the samples may be attributed to environmental or cultural effects. The paper concludes that there is evidence that cultural differences affect the views of respondents in a variety of measurable ways including in their opinion of ecumenical cooperation and in their attitudes to the poor or disadvantaged

Dual roles for LUBAC signaling in thymic epithelial cell development and survival

Thymic epithelial cells (TECs) form a unique microenvironment that orchestrates T cell differentiation and immunological tolerance. Despite the importance of TECs for adaptive immunity, there is an incomplete understanding of the signalling networks that support their differentiation and survival. We report that the linear ubiquitin chain assembly complex (LUBAC) is essential for medullary TEC (mTEC) differentiation, cortical TEC survival and prevention of premature thymic atrophy. TEC-specific loss of LUBAC proteins, HOIL-1 or HOIP, severely impaired expansion of the thymic medulla and AIRE-expressing cells. Furthermore, HOIL-1-deficiency caused early thymic atrophy due to Caspase-8/MLKL-dependent apoptosis/necroptosis of cortical TECs. By contrast, deficiency in the LUBAC component, SHARPIN, caused relatively mild defects only in mTECs. These distinct roles for LUBAC components in TECs correlate with their function in linear ubiquitination, NFκB activation and cell survival. Thus, our findings reveal dual roles for LUBAC signaling in TEC differentiation and survival

Non-Alcoholic Fatty Liver Disease and Vitamin D in the UK Biobank: A Two-Sample Bidirectional Mendelian Randomisation Study

Evidence for a role for vitamin D in non-alcoholic fatty liver disease (NAFLD) pathogenesis is conflicting. As Mendelian randomisation (MR) avoids many limitations of conventional observational studies, this two-sample bidirectional MR analysis was conducted to determine the following: (i) whether genetically predicted 25-hydroxyvitamin D [25(OH)D] levels are a risk factor for NAFLD, and (ii) whether genetic risk for NAFLD influences 25(OH)D levels. Single-nucleotide polymorphisms (SNPs) associated with serum 25(OH)D levels were obtained from the European ancestry-derived SUNLIGHT consortium. SNPs associated with NAFLD or NASH (p-value < 1 × 10−5) were extracted from previous studies and supplemented by genome-wide association studies (GWASs) performed in the UK Biobank. These GWASs were done both without (primary analysis) and with (sensitivity analysis) the population-level exclusion of other liver diseases (e.g., alcoholic liver diseases, toxic liver diseases, viral hepatitis, etc.). Subsequently, MR analyses were performed to obtain effect estimates using inverse variance weighted (IVW) random effect models. Cochran’s Q statistic, MR-Egger regression intercept, MR pleiotropy residual sum and outlier (MR-PRESSO) analyses were used to assess pleiotropy. No causal association of genetically predicted serum 25(OH)D (per standard deviation increase) with risk of NAFLD was identified in either the primary analysis: n = 2757 cases, n = 460,161 controls, odds ratio (95% confidence interval): 0.95 (0.76, −1.18), p = 0.614; or the sensitivity analysis. Reciprocally, no causal association was identified between the genetic risk of NAFLD and serum 25(OH)D levels, OR = 1.00 (0.99, 1.02, p = 0.665). In conclusion, this MR analysis found no evidence of an association between serum 25(OH)D levels and NAFLD in a large European cohort

Thermal separation of 99mTc from Molybdenum targets

Thermal separation is defined as a mass transfer process driven by molecular forces. The process involves the heat transfer between two phases with different composition. In general, thermal separation occurs when heat is generated in the system additionally to the already existing phases. In a second phase the mass is transferred in the system (adsorption) and at the end of this step the separation is completed. The thermal separation can be achieved in temperature or concentration gradient function of system configuration [1]. Thermo-chromatography is a process in which the separation occurs in gase-ous phase. By passing a heated gas through a column a thermal gradient is created with a continuously decreasing temperature along the column. The separation occurs based on the different volatilization temperatures, the less volatile species will condense on the column walls at the higher temperatures and the highly volatile compounds will condense at lower temperatures. Parameters like temperature, carrier flow rate, column geometry and length have impact on the absorption of the compound on the column material affecting the separation efficiency. The thermal separation has been used for separation of Molybdenum (Mo) and Technetium (Tc) by either sublimation in the case of 94mTc {2,3,4] or dry distillation in the case of 99mTc from neutron irradiated MoO3 [5]. The thermal separation process has been used in the development of a new type of Mo/Tc generators starting from the MoO3 as target material for production of 99mTc in linear accelerators [6]. Dry distillation has become a standard procedure for separation of radioiodine from tellurium targets [7]. The present paper describes the thermal separation of a three component system (Cu/Mo/Tc) used as a target in the production of 99mTc through the 100Mo(p,2n) reaction. Material and Methods The separation method involves the use of oxygen as a carrier gas and oxidation agent. The method is based on the different volatilization temperatures of Tc formed oxides and the MoO3 formed in the system during the oxidation. In the presence of oxygen the existing Tc is oxidized to its anhydride as Tc2O7 (b.p. 319 ⁰C; m.p. 110.9 ⁰C) following the reaction: 4Tc + 7O2 →2Tc2O7 The T2O7 has a saturated vapor pressure of 310 ⁰C whilst Mo is completely oxidized to MoO3 having a sublimation temperature at 750 ⁰C. The initial experimental setup comprised a quartz tube (6 mm internal diameter, 40 cm long) which is introduced into a horizontal tube furnace (model 55035A, Lindberg). The left end of the quartz tube is connected to a pure oxygen supply which flows through the separation tube at a rate of 10 mL/min. The other end of the tube is opened to the atmosphere and protected with quartz wool. The quartz tube is heated over a length of 23 cm at a temperature of 850 ⁰C. The heated carrier gas is flowing on the tube length and the temperature gradient is created along the tube from 850 ⁰C to room temperature. During the process, the oxygen carries out the Tc oxides to a lower temperature and Tc2O7 is deposited in the cooler region of the tube in a similar manner as described by Tachimory [5]. The temperature gradient is calibrated by meas-uring the temperature inside the tube at each centimeter along its length (FIG. 1). The radioactivity counting is performed by scan-ning the tube along its length every 2 centimeters by using a detection system shown in figure 2. The system comprises a GM tube coupled to a computer controlled linear actuator (Velmex Unislide). The tube is placed at a distance of approximately 25 mm from the collimator of GM. Preliminary testing using Mo powder Prior to testing the three component separation, a reference test was performed by using 120 mg of natural Mo powder (Alpha Aesar, 99.9 %) soaked with 50 MBq NaTcO4 (Cardinal Health, radiochemical purity >95 %). After evaporation the dried powder was introduced into a quartz tube (6 mm ID, 40 mm long) and heated up to 850 ⁰C in the presence of oxygen flowing at a rate of 10 mL/min. Three component separation The targets prepared for the production of 99mTc by a cyclotron were comprised of copper (Cu) (C101, oxygen free) support having a Mo layer deposited on the surface in an elliptical form as described in literature [8,9]. About 60 to 250 mg of Mo (99.9%, Alpha Aesar) was deposited on the target surface. 70 MBq of Tc (Cardinal Health) as NaTcO4 (> 99 % radiochemical purity) was deposited on the Mo insert to mimic the conditions created during proton irradiation. The Tc spike was evaporated to dryness and the Cu/Mo/Tc target was then introduced into the experimental setup. The process was allowed to continue for 20 min. The experiment was carried out by inserting the target plates in a quartz tube (CanSci, Canada) of similar design to those described by Fonslet for the separation of radio-iodine from TeO2 targets [7]. The quartz tube can be seen in FIG. 2 and illustrated with dimensions indicated in FIG. 3. Separation of in-situ cyclotron produced Tc by irradiation of Mo targets with a proton beam. A third set of experiments have been performed for in-situ generated Tc by irradiation of circular targets containing approximately 60 mg Mo deposited on a copper support. The targets were irradiated for 30 min with a proton beam with the energy of 15 MeV and a current of 50 µA. The separation was performed using similar experimental conditions as previously described. The quartz tube was scanned in length by using a RadioTLC scanning system calibrated for 99mTc and 99Mo isotopes. After the thermal separation was completed 99mTc was recovered as NaTcO4 by selectively washing the quartz tube with 1 M NaOH (Fisher) solution. The presence of Mo in the NaTcO4 solution was verified by a colorimetric strip test (EM-Quant Mo test kit, Millipore). The presence of copper was qualitatively analyzed by adding a few drops of concentrated NH4OH (Fisher) solution and checking the formation of Schweitzer reagent. Results Thermal separation of Tc-Mo powder After 20 min the deposition of MoO3 was ob-served as yellow crystals in the region of tem-perature of 770 ⁰C, which is in accordance with the results reported in the literature [5]. The activity of 99mTc was detected at about 5 cm from the exit of the tube furnace in a temperature range starting with 310 ⁰C and ending at 46 ⁰C (FIG. 4)

Skeletal adaptations in young male mice after 4 weeks aboard the International Space Station

Gravity has an important role in both the development and maintenance of bone mass. This is most evident in the rapid and intense bone loss observed in both humans and animals exposed to extended periods of microgravity in spaceflight. Here, cohabitating 9-week-old male C57BL/6 mice resided in spaceflight for ~4 weeks. A skeletal survey of these mice was compared to both habitat matched ground controls to determine the effects of microgravity and baseline samples in order to determine the effects of skeletal maturation on the resulting phenotype. We hypothesized that weight-bearing bones would experience an accelerated loss of bone mass compared to non-weight-bearing bones, and that spaceflight would also inhibit skeletal maturation in male mice. As expected, spaceflight had major negative effects on trabecular bone mass of the following weight-bearing bones: femur, tibia, and vertebrae. Interestingly, as opposed to the bone loss traditionally characterized for most weight-bearing skeletal compartments, the effects of spaceflight on the ribs and sternum resembled a failure to accumulate bone mass. Our study further adds to the insight that gravity has site-specific influences on the skeleton