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Background: COVID-19 pandemic resulted in a death of 419 patients among total admissions of 10682 with a death rate of 3.92% in the tertiary care COVID-19 hospital. We studied the biochemical and hematological parameters among 241 patients who died of the disease. Results: CRP values were raised above 12mg/L in 58% of patients. 83% of patients had elevated LDH levels of >250 IU/L. Procalcitonin levels were above 0.5 microgram/L in nearly 66% of patients. Serum ferritin was more than 500 micrograms/L in 51% of patients. Elevated IL-6 were found in 83% of patients making it a significant inflammatory parameters. D-dimer levels were more than 500ng/ml in 74% of patients. HS Troponin I was raised in 83% of patients. Leukocytosis of more than 11000/Cu mm was seen in 38%. Leukopenia was seen in 35%. Thrombocytopenia was seen in 27% and normal platelet count was seen in 62%. Conclusions: Biochemical parameters help in assessing the severity of inflammation in COVID-19 disease. They aid in the process of treatment particularly anticoagulants and corticosteroid. Specific parameters like IL-6 can help in decision making by treating physician regarding the use of anti IL-6 drugs like Tocilizumab. Elevated HS troponin I in our study showed myocardial injury played a significant role in mortality of COVID 19 patients at our centre. Keywords: Creactive protein (CRP), Ferritin, Ddimer, Procalcitonin (PCT), Lactic dehydrogenase (LDH), IL6, HS Tropanin I.

Background: Induction of labour is the artificial initiation of labour before the spontaneous onset, and it is the common obstetric intervention. Aim: To compare the potency of oral misoprostol solution with vaginal misoprostol to induce labour at term. Settings and Design: This randomized control study was carried out from July 2020 to September 2020 at Government General Hospital, Kakinada. Materials and Methods: 80 women requiring induction were recruited in the study. The women were randomized to receive 20 micrograms of oral misoprostol solution every 2nd hourly or vaginally 25 micrograms of misoprostol every 6th hourly until the labour sets in the active phase of labour. Delivery within 24 hours was the outcome in the study. Statistical Analysis: Data were documented in Microsoft excel and analyzed using SPSS software. Results: 82.5% of women in the oral group delivered by spontaneous vaginal delivery, whereas 75% in the vaginal group. In the oral group, 37.5% of delivery occurred within 6-12 hours, 40% delivered within 12-18 hours duration, whereas in the vaginal group, 12.5% delivered within 6-12hours duration, 50% delivered within 12-18 hours duration. Oxytocin augmentation was required in 25% of the oral group, whereas 42.5% in the vaginal group. Meconium stained liquor was 12.5% in the oral group, whereas 20% in the vaginal group.12.5% underwent cesarean section in the oral group, 10% in the vaginal group. Conclusion: Compared with vaginal misoprostol, oral misoprostol solution results in shorter induction delivery interval and less oxytocin augmentation required. Keywords: Induction of labour, Oral misoprostol solution, Per vaginal misoprostol.

The presence of antibodies against endemic coronaviruses has been linked to disease severity after SARS-CoV-2 infection. Assays capable of concomitantly detecting antibodies against endemic coronaviridae such as OC43, 229E, NL63, and SARS-CoV-2 may help to elucidate this question. We developed a serum screening platform using a bead-based Western blot system called DigiWest, capable of running hundreds of assays using microgram amounts of protein prepared directly from different viruses. Characterization of the immunoassay for detection of SARS-CoV-2 specific antibodies revealed a sensitivity of 90.3% and a diagnostic specificity of 98.1%. Concordance analysis with the SARS-CoV-2 immunoassays available by Roche, Siemens, and Euroimmun indicates comparable assay performances (Cohen's κ ranging from 0.8874 to 0.9508). Analogous assays for OC43, 229E, and NL63 were established and combined into one multiplex with the SARS-CoV-2 assay. Seroreactivity for different coronaviruses was detected with high incidence, and the multiplex assay was adapted for serum screening.

Background The prognostic significance of serum biomarkers in patients with intracerebral hemorrhage (ICH) is not well investigated concerning inhospital mortality (IHM) and cardiopulmonary events within the first 24 hours of intensive care unit (ICU) treatment. The influence of troponin I (TNI) value and cortisol value (CV) on cardiopulmonary events within the first 24 hours of ICU treatment was reported in subarachnoid hemorrhage patients, but not in ICH patients up to now. The aim of this study was to investigate the role of early serum biomarkers on IHM and TNI value and CV on cardiopulmonary events within the first 24 hours of ICU treatment. Patients and Methods A total of 329 patients with spontaneous ICH were retrospectively analyzed. Blood samples were taken on admission to measure serum biomarkers. The TNI value and CV were defined as biomarkers for cardiopulmonary stress. Demographic data, cardiopulmonary parameters, including norepinephrine application rate (NAR) in microgram per kilogram per minute and inspiratory oxygen fraction (FiO2) within the first 24 hours, and treatment regime were analyzed concerning their impact on ICU treatment and in hospital outcome. Binary logistic analysis was used to identify independent prognostic factors for IHM. Results Patients with initially nonelevated CVs required higher NAR (p = 0.01) and FiO2 (p = 0.046) within the first 24 hours of ICU treatment. Lower cholinesterase level (p = 0.004), higher NAR (p = 0.002), advanced age (p Conclusion Higher levels of C-reactive protein, serum lactate, blood glucose, and lower cholinesterase level on admission were significantly associated with IHM. Patients with initially nonelevated CVs required higher NAR and FiO2 within the first 24 hours of ICU treatment. Furthermore, requiring an NAR > 0.5 µg/kg/min or an FiO2 > 0.21 were identified as additional independent predictors for IHM. These results could be helpful to improve ICU treatment in ICH patients.

Thyroid stimulating hormone is the main hormone and 80 microgram is secreted per day. Ageing changes occur in all body systems including endocrine system. In elderly individuals the non-specific clinical manifestations of hypo and hyperthyroidism has confusion in the clinical setup. In females, up to the age of 30, there is a decrease in TSH levels thereafter there is a slight but definite increase in TSH levels. In males, there is a slight increase in TSH with age up to 90 years. Thyroid treatment in the elderly should take into consideration symptoms and TSH values. Key words: TSH, Thyroid changes in elderly, Effect of ageing on TSH, Thyroid hormone.

Multiple consensus statements have called for preclinical randomized controlled trials (pRCT) to improve translation in stroke research. Here, we investigated the efficacy of IL-17A neutralizing antibodies in a multicentric pRCT using a murine stroke model. C57/Bl.6 mice were subjected to transient middle cerebral artery occlusion (tMCAO). Mice were randomly allocated (1:1). Either anti-IL-17A (500 µg) or isotype antibody (500 µg) were administered 1 h after tMCAO. Primary analysis of infarct volumes was done by MRI after three days. Secondary analysis included mortality, neurological score, neutrophil infiltration and the impact of the gut microbiome on treatment effects. Out of 136 mice, 109 mice were included in the analysis. Mixed model analysis revealed that the IL-17A neutralization significantly reduced infarct sizes (anti IL-17A: 61.77 ± 31.04 mm3; IgG control: 75.66 ± 34.79 mm3; p=0.01). Secondary outcome measures showed a decrease in mortality (Hazard Ratio=3.43, 95% CI = 1.157 - 10.18; p=0.04) and neutrophil invasion into ischemic cortices. There was no difference in the neurological score. The analysis of the gut microbiome showed significant differences between centers. Taken together, this is the first positive pRCT in an ischemia reperfusion model. It suggests IL-17A neutralization as a potential target in stroke.

International audience; This study explores the potential of profiling a microgram-scale soft tissue biopsy by NMR spectroscopy. The important elements of high resolution and high sensitivity for the spectral data are achieved through a unique probe, HR-μMAS, which allowed comprehensive profiling to be performed on microgram tissue for the first time under MAS conditions. Thorough spatially resolved metabolic maps were acquired across a coronal brain slice of rat C6 gliomas, which rendered the delineation of the tumor lesion. The results present a unique ex vivo NMR possibility to analyze tissue pathology that cannot be fully explored by the conventional approach, HR-MAS and in vivo MRS. Aside from the capability of analyzing a small localized region to track its specific metabolism, it could also offer the possibility to carry out longitudinal investigations on live animals due to the feasibility of minimally invasive tissue excision.

Autoimmune hypophysitis is an autoimmune inflammation of the pituitary gland. It may result in hypopituitarism depending upon the part of pituitary affected. Here, we report the case of autoimmune hypophysitis in a 30-year-old male who was admitted with complaints of generalised swelling for one month and fever and cough for one week. The patient’s Thyroid-stimulating hormone (TSH) was more than 139, anti-thyroid peroxidase antibody (anti-TPO) was more than 1000 and serum sodium was 107 and a tiny streak of tissue along the periphery with an empty sella as seen on MRI. The patient was given tablet Levothyroxine 100 microgram, Telmisartan 40 mg and Hydrocortisone 10mg twice daily. With medications, the patient improved and was discharged.

Introduction: Granulocyte colony stimulating factor improves short-term survival and clinical outcomes in alcoholic hepatitis, acute-on-chronic liver failure and decompensated chronic liver disease. Our study aimed to assess survival benefit and change in Child-Turcotte-Pugh and Model For End-Stage Liver Disease scores 30 days after Granulocyte colony stimulating factor therapy in chronic liver disease patients, irrespective of their mode of presentation. Methods: This was a prospective observational study conducted in a university teaching hospital, where 25 patients with chronic liver disease were given 300 micrograms of Granulocyte colony stimulating factor subcutaneously 12 hourly plus standard medical therapy. We assessed survival until day 30. Child-Turcotte- Pugh and Model For End-Stage Liver Disease scores at enrolment and 30 days after treatment were compared. Results: 21 of 25 patients treated with Granulocyte colony stimulating factor survived at day 30. Treatment with Granulocyte colony stimulating factor reduced Child-Turcotte-Pugh score from 10.33 ± 1.24 to 8.76 ± 1.79 (p< 0.001) at day 30 and Model For End-Stage Liver Disease score from 22.10 ± 4.67 to 16.38 ± 5.52 (p < 0.001) at day 30. Conclusions: Granulocyte colony stimulating factor improves clinical outcome, Child-Turcotte-Pugh and Model For End-Stage Liver Disease scores in patients admitted with chronic liver disease for any cause. Further studies are needed to explore whether lower doses (total six doses) of Granulocyte colony stimulating factor are as effective as higher doses (total 10 doses).

Background: Quantitation of Helicobacter pylori (Hp) in the gastric tissue is essential for assessment of vaccination/therapeutic regimens. Methods & Results: Here, the inhibitory effect of mouse gastric DNA (MgDNA) on amplification of Hp genomic DNA (HpDNA) was evaluated by spiking HpDNA with serial dilutions of MgDNA, which yielded concentrations of >10 ng/µl and >0.63-10 ng/µl of MgDNA, as inhibition and interference zones, respectively. Mice were then inoculated with varying doses of Hp and assessed at the inhibition-free concentration of 0.63 ng/µl. The average Hp copy numbers per microgram of gastric tissue discriminated mice having received high vs. low dose inoculums (p < 0.001). Secondly, Hp copy numbers were quantitated in immunized mice, which demonstrated significantly lower numbeZrs, in reference to controls (p = 0.006). Conclusion: Our method, bypassing the inhibition and/or interference imposed by MgDNA, was able to quantitate gastric tissue-colonizing Hp, segregating mice inoculated with low vs. high doses of Hp, as well as those immunized from controls.

Aim Compare the effectiveness of administration of different doses of rectal misoprostol before cesarean section to reduce intra- and postoperative blood loss. Methods A double-blind randomized clinical trial including 453 term pregnant woman scheduled for elective cesarean section where participants received either 200-, 400- or 600-μg misoprostol rectally before cesarean section. Study medications were administered after catheter insertion and shortly before skin incision. Primary outcome measures were intraoperative blood loss. Results The intraoperative blood loss was higher in patients who received 200-μg misoprostol (464.6 ± 143.1 mL) than those who received 400 or 600 μg, yet, no statistical difference was found between the 400- (359.3 ± 120.9 mL) and 600-μg groups (330.8 ± 133.8 mL). The incidence of side effects as fever and chills increases with increasing the dose of misoprostol. Conclusion Rectal administration of misoprostol for the prevention of post-partum hemorrhage and decreasing intraoperative blood loss during caesarian section is a good alternative to other uterotonics. Yet, the best dose to be used needs further research to be agreed upon.

International audience

Developing solid oral drug products with good content uniformity (CU) at low doses is challenging; this challenge further aggravates when the tablet size decreases from a conventional tablet to a micro/mini-tablet (1.2–3 mm diameter). To alleviate the CU issues, we present a novel use of nanocrystalline suspension combined with high shear wet granulation for the first time. In this approach, nanomilled drug in the form of nanocrystalline suspension is sprayed onto the powder bed to ensure uniform distribution. The resulting granules had adequate particle size distribution and flow characteristics to enable manufacturing of micro-tablets with good weight uniformity and tensile strength. Nanomilled drug resulted in excellent content uniformity among individual micro-tablets even at a dose strength as low as 0.16 mcg, whereas micronized drug resulted in unacceptable CU even at 5x higher dose strength (0.8 mcg). Besides, the use of nanomilled drug has enhanced the dosing flexibility of micro-tablets and showed superior dissolution performance in comparison with micronized drug with no impact of storage conditions (40 °C/75%RH for six months) on their dissolution performance. The proposed approach is simple and can be easily incorporated into traditional high shear wet granulation process to develop sub-microgram dose solid oral drug products.

Background In the absence of a specific drug for COVID 19, treatment with plant extracts could be an option worthy of further investigation and has motivated to evaluate the safety and anti-SARS-CoV-2 activity of plant extracts. Purpose To screen the phytochemicals for anti-SARS-CoV-2 in silico and evaluate their safety and efficacy in vitro and in vivo. Method The phytochemicals for anti-SARS-CoV-2 were screened in silico using molecular docking. The hits generated from in silico screening were subjected for extraction, isolation and purification. The anti-SARS-CoV-2 activity of Zanthoxylum piperitum (E1), Withania somnifera (E2), Calophyllum inophyllum (E3), Andrographis paniculata (E4), Centella asiatica (E5) ethanol extracts. The aerial parts were used for E1, E3, E4, E5 and root was used for E2. The in vitro safety and anti-SARS-CoV-2 activity of plant methanol extracts were performed in VeroE6 cells using Remdesivir as positive control. The acute and sub-acute toxicity study was performed in Wistar male and female rats. Results The percentage of cell viability for E4, E5 and E2 treated VeroE6 cells were remarkably good on the 24th and 48th hour of treatment. The in vitro anti-SARS-CoV-2 activity of E4, E5 and E2 were significant for both E gene and N gene. The percentage of SARS-CoV-2 inhibition for E4 was better than Remdesivir. For E gene and N gene, Remdesivir showed IC50 of 0.15 µM and 0.11 µM respectively, For E gene and N gene, E4 showed IC50 of 1.18 µg and 1.16 µg respectively. Taking the clue from in vitro findings, the E4, E5 and E2 were combined (E 4.5.2) and evaluated for acute and sub-acute toxicity in Wistar male and female rats. No statistically significant difference in haematological, biochemical and histopathological parameters were noticed. Conclusion The study demonstrated the anti-SARS-CoV-2 activity in vitro and safety of plant extracts in both in vitro and in vivo experimental conditions., Graphical abstract Image, graphical abstract

BackgroundSeroreactivity against human endemic coronaviruses has been linked to disease severity after SARS-CoV-2 infection. Assays that are capable of concomitantly detecting antibodies against endemic coronaviridae such as OC43, 229E, NL63, and SARS-CoV-2 may help to elucidate this question. We set up a platform for serum-screening and developed a bead-based Western blot system, namely DigiWest, capable of running hundreds of assays using microgram amounts of protein prepared directly from different viruses.MethodsThe parallelized and miniaturised DigiWest assay was adapted for detecting antibodies using whole protein extract prepared from isolated SARS-CoV-2 virus particles. After characterisation and optimization of the newly established test, whole virus lysates of OC43, 229E, and NL63 were integrated into the system.ResultsThe DigiWest-based immunoassay system for detection of SARS-CoV-2 specific antibodies shows a sensitivity of 87.2 % and diagnostic specificity of 100 %. Concordance analysis with the SARS-CoV-2 immunoassays available by Roche, Siemens, and Euroimmun indicates a comparable assay performance (Cohen’s Kappa ranging from 0.8799-0.9429). In the multiplexed assay, antibodies against the endemic coronaviruses OC43, 229E, and NL63 were detected, displaying a high incidence of seroreactivity against these coronaviruses.ConclusionThe DigiWest-based immunoassay, which uses authentic antigens from isolated virus particles, is capable of detecting individual serum responses against SARS-CoV-2 with high specificity and sensitivity in one multiplexed assay. It shows high concordance with other commercially available serologic assays. The DigiWest approach enables a concomitant detection of antibodies against different endemic coronaviruses and will help to elucidate the role of these possibly cross-reactive antibodies.

The aim of this research is to reduce the number of peroxide in two frying oil samples (Aldura and altusan) by using one of the natural antioxidants which is vitamin E. one of the chemical specifications that assess the oil is the peroxide number, which is the amount of free radicals formed in the oil and the number of peroxides increases during exposure to heat, moisture, oxygen, bad storage conditions, light or other conditions. These peroxides have a harmful effect on the consumer health (initiation cancer). The vitamin E had a clear impact on the peroxide value, as when conducting the experiment and using oil to fry potato slices, the number of peroxide was high before adding vitamin E which score (Peroxide number is 5.6 for 15 minutes, 9.8 for 30 minutes, and 11.6 for 45 minutes). After using vitamin E in amount of 10 microgram per 100 ml of oil, the effect was clear of vitamin E on the number of peroxide as it decreased significantly. Which scored (5 for 15 minutes, 8.1 for 30 minutes, 8.9 for 45 minutes). When vitamin E added by 20 microgram the result was (4.8 for 15 minutes, 5.5 for 30 minutes, 6.5 for 45 minutes). 50 microgram per 100 oil shown to have no significant effect on the peroxide value. it was found that the best concentration is 20 microgram for a sample of 100 ml of oil.

Objectives: To the best of our knowledge, there is no published study regarding use of IFN beta-1a in the treatment of severe COVID-19. In this randomized clinical trial efficacy and safety of IFN β-1a has been evaluated in patients with severe COVID-19. Methods: Forty-two patients in the interferon group received IFN beta-1a in addition to the standard of care. Each 44 micrograms/ml (12 million IU/ml) of interferon beta-1a was subcutaneously injected three times weekly for two consecutive weeks. The control group received only the standard of care. Primary outcome of study was time to reach clinical response. Secondary outcomes duration of hospital stay, length of ICU stay, 28-day mortality, effect of early or late administration of IFN on mortality, adverse effects and complications during the hospitalization. Results: As primary outcome, time to the clinical response was not significantly different between the IFN and the control groups (9.7 +/- 5.8 vs. 8.3 +/- 4.9 days respectively, P=0.95). On day 14, 66.7% vs. 43.6% of patients in the IFN group and the control group were discharged, respectively (OR= 2.5; 95% CI: 1.05- 6.37). The 28-day overall mortality was significantly lower in the IFN then the control group (19% vs. 43.6% respectively, p= 0.015). Early administration significantly reduced mortality (OR=13.5; 95% CI: 1.5-118). Conclusion: Although did not change time to reach the clinical response, adding to the standard of care significantly increased discharge rate on day 14 and decreased 28-day mortality. Clinical Trial Registration ID #IRCT20100228003449N28.

Leishmaniasis is a widespread tropical infection; cutaneous leishmaniasis (CL) is the most common form of this disease known to cause significant morbidity. Trace metals, including selenium, zinc, iron, and copper, are required for the activity of several enzymes involved in immune system responses. The aim of this research was to measure the serum levels of selenium (Se), zinc (Zn), copper (Cu), iron (Fe), and Zn/Cu ratio in patients with CL. In this case-control study, 80 patients with CL and 80 healthy volunteers (not exposed to CL) from a CL endemic region in southwest Iran agreed to participate. Both clinical and parasitological verifications were made to include each subject as a CL-positive case. A questionnaire was completed for each participant which included the following criteria: age (year), height (cm), weight (kg), body mass index (kg/m2), and duration of disease (day). The biochemical assays were performed according to the standard protocols, and the values of Zn, Cu, Se, and Fe were expressed in micrograms per deciliter (μg/dl). All results were expressed as mean ± standard deviation (SD), and the statistical significance level was defined to be less than 0.05 (P 0.05). The mean ± SD concentrations of Zn, Fe, and Se in the control group were found to be 118.87 ± 6.35 μg/dl, 123.00 ± 8.40 μg/dl, and 11.26 ± 1.88 μg/dl, respectively. These trace elements (TEs) were statistically lower (P < 0.001) in patients with CL (case group) with values of 83.05 ± 7.32 μg/dl for Zn, 86.51 ± 10.09 μg/dl for Fe, and 3.83 ± 1.20 μg/dl for Se. We have also observed that serum levels of Cu in CL-positive group were significantly higher than in the controls (P < 0.001). Furthermore, CL patients had significantly lower Zn/Cu ratio than controls (0.63 ± 0.05 μg/dl vs. 1.11 ± 0.10; P < 0.001). The alternation in serum levels of TEs may be a part of the defense strategy of the organism. Based on these results, it can be suggested that serum levels of these TEs can be a useful marker to estimate the prognosis of CL infection.

Background Anaemia is a common complication of advanced disease with associated symptoms having a negative impact on quality of life. A recent national hospice audit suggested a need for improvement in the management of anaemia in hospices, with a particular focus on investigation and more restrictive use of blood. As a result, we have updated local hospice guidelines and reviewed their use to assess the impact on patient care. Method Alongside the introduction of updated guidelines, prospective data were collected from hospice inpatients with symptoms related to anaemia, who were being investigated for potential treatment. The data, collected over three months, included blood parameters to assess cause of anaemia and guide treatment options. Results Of the seven patients investigated, four were folate deficient and five had either iron deficiency or functional iron deficiency. Two were treated with folate replacement; one subsequently deteriorated rapidly. One was too unwell to be treated and the folate level improved in one without treatment. Four out of six patients (one not measured) had reduced serum iron yet ALL ferritin results were >30 microgram/L and five had a ferritin >1000 microgram/L (range 186–3383). CRP results ranged from 95 to 206. Conclusion This case series highlights the complexity of interpreting results to guide management of anaemia in the hospice inpatient population. When a deficiency is identified, treatment with folic acid or iron carries less risk than treatment with blood. However, with ferritin levels this high, treatment with oral or intravenous iron is not usually recommended. Iron may not represent a frequently useful treatment option in this population. We are not aware of evidence confirming a positive clinical response to these treatments in hospice patients with complex underlying aetiologies contributing to anaemia. We believe this highlights a need for research to demonstrate clinical benefit to support these treatments.

Stable isotope analysis of biogenic carbonates has remained one of the most important tools in paleoceanography since Emiliani (1955) first described the fluctuation of oxygen isotopes in planktic foraminifers over the Pleistocene. Many laboratories now possess equipment with the capability to analyse foraminifer specimens singularly, at least for larger planktic forms.Being able to run single specimens of planktic foraminifers is significant, because it yields entirely different information than when one would analyse multiple specimens from the same species. Planktic foraminifers have an average life span of about one month, so analysing single specimens, makes paleoceanographic data at seasonal resolution available (e.g. Ganssen et al., 2011; Metcalfe et al 2019, and references therein).Most modern equipment for stable isotope analysis of CaCO3 samples yields good precision down to 10 microgram sample size. The smallest samples are generally measured with a dual inlet technique, because that quantitatively collects the CO2 gas sample in a cold trap before analysis, leading to a more efficient use of the sample gas. Modern dual inlet equipment has a sample size limit somewhere between 10 and 6 microgram CaCO3 sample weight, and in that range usually operates at increased analytical uncertainty when compared to larger samples (e.g. Ganssen et al., 2011). Smaller samples are problematic, because at small amounts of sample gas, the dual inlet system is not able to maintain viscous flow conditions required for precise isotope analysis. To circumvent that barrier, one can use continuous-flow (CF) mass spectrometry because in CF systems the carrier gas ensures proper flow conditions even if there is (virtually) no sample gas produced. Doing so has previously allowed for the isotope analysis of CaCO3 samples in the 10 – 6 microgram range at an external precision (1SD) of ~0.12‰ for both δ18O and δ13C (e.g. Metcalfe et al 2019).To further improve the performance of CF mass spectrometry for small CaCO3 samples, we ran experiments on a Thermo GASBENCH system, equipped with a cold trap (cf. Fiebig et al 2005) and interfaced with a Delta-V mass spectrometer. The experiments consisted of replicate analysis of CaCO3 standards between 10 and 3 micrograms in weight, which is the weight range of many of the smaller specimens of planktic foraminifers.Several hardware modifications were implemented to improve system stability and remove observed effects of contribution of blank CO2 building up in the sample vials. With these modifications, external reproducibility of the set-up for carbonate standard aliquots between 10 and 4 microgram reached a precision of ~0.10 ‰ for both δ18O and δ13C (1SD). This is similar to precisions typically attained for routine analysis of much larger samples in standard operation on the same equipment, and demonstrates that precise stable isotope analysis of smaller single-specimen planktic foraminifers than we could achieve so far is within reach of CF mass spectrometry.References:Emiliani, C. 1955, DOI: 10.1086/626295Fiebig, J., et al. 2005, DOI: 10.1002/rcm.2060Ganssen, G.M., et al., 2011, DOI:10.5194/cp-7-1337-2011Metcalfe, B., et al., 2019, DOI: 10.1029/2018PA003475

Recombinant adeno-associated virus (rAAV)rh74.MCK.GALGT2 is a muscle-specific gene therapy that is being developed to treat forms of muscular dystrophy. Here we report on an isolated limb infusion technique in a non-human primate model, where hindlimb blood flow is transiently isolated using balloon catheters to concentrate vector in targeted leg muscles. A bilateral dose of 2.5 × 1013 vector genomes (vg)/kg/limb was sufficient to induce GALGT2-induced glycosylation in 10%–60% of skeletal myofibers in all leg muscles examined. There was a 19-fold ± 6-fold average limb-wide increase in vector genomes per microgram genomic DNA at a bilateral dose of 2.5 × 1013 vg/kg/limb compared with a bilateral dose of 6 × 1012 vg/kg/limb. A unilateral dose of 6 × 1013 vg/kg/limb showed a 12- ± 3-fold increase in treated limb muscles compared to contralateral untreated limb muscles, which received vector only after release into the systemic circulation from the treated limb. Variability in AAV biodistribution between different segments of the same muscle was 125% ± 18% for any given dose, while variability between the same muscle for any given treatment dose was 45% ± 7%. These experiments demonstrate that treatment of muscles throughout the leg with rAAVrh74.MCK.GALGT2 can be accomplished safely using an isolated limb infusion technique, where balloon catheters transiently isolate the limb vasculature, but that intra- and inter-muscle transduction variability is a significant issue. Keywords: limb perfusion, gene therapy, Galgt2, muscular dystrophy

BackgroundA novel murine bi-functional molecule, G04-trap, comprised of an anti-CD73 antibody fused to the extracellular domain of TGFβ receptor II, is designed to potently antagonize two prominent immunosuppressive and pro-tumorigenic pathways present across a variety of cancer types. Inhibition of both CD73-adenosine and TGFβ pathways is expected to create favorable conditions within the tumor microenvironment and restore antitumor immune responses.MethodsG04-trap was evaluated in Detroit562, MC38, and Hepa1-6 efficacy tumor models. Tumor growth inhibition (TGI) was determined when >/=9 animals were alive in each group. Tumor-bearing mice received isotype control (200 microgram), G04-trap (246 microgram), anti-PD-(L)1 (200 microgram) or G04-trap + anti-PD-(L)1 twice per week for 3 weeks. Pharmacokinetic (PK) and pharmacodynamic (PD) assessment was performed on MC38 tumor-bearing mice dosed with 3 mg/kg, 10 mg/kg, or 30 mg/kg G04-trap. Plasma and tumor PK, CD73 target occupancy on T cells, plasma TGFβ, plasma free-sCD73, and tumor CD73 activity were measured after a single dose administration of G04-trapResultsAdministration of G04-trap to mice harboring TGFβ-dependent human pharyngeal Detroit562 xenograft tumors led to a dose-dependent anti-tumor response (83% TGI, at 246 microgram vs. isotype control on day 21). In addition, treatment with G04-trap in combination with immune checkpoint inhibition showed anti-tumor activity in MC38 and Hepa1-6 syngeneic mouse models. In MC38 on day 18, there was a statistically significant TGI with G04-trap + anti-PD-L1 (99% TGI vs. isotype control or 98% TGI vs. anti-PD-L1 alone). A more modest effect was observed in Hepa1-6, with 47% TGI in mice receiving G04-trap + anti-PD-1 vs. isotype control on day 27. To further interpret the efficacy observed in the MC38 tumor model, we performed in-depth PK/PD analysis. Intravenous administration of G04-trap at 3-30 mg/kg resulted in 10% tumor-to-plasma exposure ratio. Full TGFβ target coverage and full CD73 target occupancy on blood T cells was sustained for >3 days, supporting a BIW dosing schedule in non-clinical studies. Treatment also resulted in a dose-dependent inhibition of CD73 activity in tumors. In contrast to cellular CD73, a dose-dependent increase in free sCD73 concentration above baseline was measured in the plasma, consistent with previous reports evaluating anti-CD73 antibodies [1].ConclusionsDual inhibition of CD73 and TGFβ in combination with immune checkpoint blockade resulted in enhanced anti-tumor activity in xenograft and syngeneic tumor models. These results suggest that further exploration of this approach is warranted.ReferencesZhao Y, Gu H, Postelnek J, DeMichele M, Yuan L, Zhang YJ, et al. Fit-for-purpose protein biomarker assay validation strategies using hybrid immunocapture-liquid chromatography-tandem-mass spectrometry platform: Quantitative analysis of total soluble cluster of differentiation 73. Anal Chim Acta. 2020;1126:144–53.

The present study was aimed at investigating the modulatory effects of folic acid (FA) on early stages of chemically induced skin cancer. For this, a two-stage model of skin tumorigenesis was employed. 7,12-Dimethylbenz(a)anthracene (DMBA, 500 nmol/100 ul of acetone) was applied topically for two weeks (twice weekly), followed by phorbol-12-myristate-13-acetate (TPA, 1.7 nmol/100 ul) twice weekly for six weeks on the depilated skin of mice, and FA was administered orally at a dose of 40 microgram/animal for 10 weeks daily. Balb/c mice were divided into four groups depending upon the treatment they received (control, DMBA/TPA, FA, and FA+DMBA/TPA). DMBA/TPA treatment led to the formation of papillomas in DMBA/TPA and FA+DMBA/TPA groups. Ornithine decarboxylase (ODC), proliferating cell nuclear antigen (PCNA), epidermal thickness, and cell count were evaluated to assess the beneficial effects in the early stages. FA exhibited its ameliorative potential as indicated by decreased epidermal thickness and cell count in FA+DMBA/TPA group when compared to DMBA/TPA group. Concomitantly, FA decreased the expression of ODC and PCNA in skin and activity of serum lactate dehydrogenase, suggesting inhibitory effects on cell proliferation and cell damage. Differential modulation in lipid peroxidation and reduced glutathione was observed in response to DMBA/TPA treatment and its intervention with FA. Although these findings suggest the inhibitory potential of FA during initial stages of murine skin cancer, detailed studies are warranted considering the ambiguous reports available in literature regarding the association of FA and cancer.

Background: High preoperative BNP levels can be a marker of higher risk for coronary artery bypass grafting. We investigated the predictive value of serum NT-proBNP levels undergoing coronary artery bypass grafting for the need for inotropic support. Methods: In this prospective study, preoperative serum NT-proBNP levels were obtained in 51 patients [80.4% (n=42) were males and 19.6% (n=9) were females] undergoing isolated coronary artery bypass grafting. The study patients were divided into three groups depending on NT-proBNP levels as low NT-proBNP (100 pg/mL, Group-2, 29.4%, n=15) group, and high NT-proBNP (>500 pg/mL, Group-3, 41.2%, n=21) group.Results: At postoperative day 0, the mean adrenalin, dopamine, dobutamine, and noradrenalin consumptions were 0.1±0.7 microgram/kg/min, 0.08±0.56 microgram/kg/min, 2.1±3.01 microgram/ kilogram/min, and 1.35±3.45 microgram/kg/min, respectively. There were no statistically significant differences between the groups in terms of the use of adrenalin (p=0.50, p>0.05), dopamine (p=0.31 p>0.05), dobutamine (p=0.59 p>0.05), and noradrenalin (p=0.24 p>0.05) at postoperative day 0. The doses of inotropic agents used at postoperative days 1 and 2 did not show significant differences between the groups for the three inotropic agents.Conclusion: Preoperative serum NT-proBNP levels in patients undergoing coronary artery bypass grafting are not associated with the need for inotropic support in the postoperative period.Cardiovasc. j. 2017; 9(2): 90-96

Quantitative assessment of key proteins that control the tumor-immune interface is one of the most formidable analytical challenges in immunotherapeutics. We developed a targeted MS platform to quantify programmed cell death-1 (PD-1), programmed cell death 1 ligand 1 (PD-L1), and programmed cell death 1 ligand 2 (PD-L2) at fmol/microgram protein levels in formalin fixed, paraffin-embedded sections from 22 human melanomas. PD-L1 abundance ranged 50-fold, from ∼0.03 to 1.5 fmol/microgram protein and the parallel reaction monitoring (PRM) data were largely concordant with total PD-L1-positive cell content, as analyzed by immunohistochemistry (IHC) with the E1L3N antibody. PD-1 was measured at levels up to 20-fold lower than PD-L1, but the abundances were not significantly correlated (r2 = 0.062, p = 0.264). PD-1 abundance was weakly correlated (r2 = 0.3057, p = 0.009) with the fraction of lymphocytes and histiocytes in sections. PD-L2 was measured from 0.03 to 1.90 fmol/microgram protein and the ratio of PD-L2 to PD-L1 abundance ranged from 0.03 to 2.58. In 10 samples, PD-L2 was present at more than half the level of PD-L1, which suggests that PD-L2, a higher affinity PD-1 ligand, is sufficiently abundant to contribute to T-cell downregulation. We also identified five branched mannose and N-acetylglucosamine glycans at PD-L1 position N192 in all 22 samples. Extent of PD-L1 glycan modification varied by ∼10-fold and the melanoma with the highest PD-L1 protein abundance and most abundant glycan modification yielded a very low PD-L1 IHC estimate, thus suggesting that N-glycosylation may affect IHC measurement and PD-L1 function. Additional PRM analyses quantified immune checkpoint/co-regulator proteins LAG3, IDO1, TIM-3, VISTA, and CD40, which all displayed distinct expression independent of PD-1, PD-L1, and PD-L2. Targeted MS can provide a next-generation analysis platform to advance cancer immuno-therapeutic research and diagnostics.

Colorectal cancer (CRC) is among the most common causes of cancer-related mortality.1 For the purpose of population-based CRC screening, faecal immunochemical tests (FIT) have been widely accepted.2 FIT can both be used in a qualitative manner, leading to either a positive or negative result, or a quantitative manner resulting in the reporting of microgram faecal haemoglobin (Hb) per gram faeces. Screenees with a FIT result above a prespecified threshold are referred for colonoscopy. A higher faecal Hb concentration is associated with a higher risk of advanced neoplasia.3 4 Most screening programmes use quantitative FIT. The results are however habitually reported in a dichotomised manner (ie, below or above a prespecified threshold). Such a dichotomised strategy subsequently misses out on countless possibilities in which the exact faecal Hb concentration could guide clinical decision-making. One of these possibilities is the use of negative FIT results, in other words faecal Hb concentrations below the accepted cut-off, as a predictor for the risk of advanced neoplasia in following screening rounds. The beautiful paper by Senore and colleagues in Gut provides additional support for this concept in exploring the predictive value of faecal Hb …

A microgram-scale X-ray structure analysis of small molecules was developed by applying a modern high-throughput crystallization technique to classical co-crystallization via acid-base salt formation. The use of an automatic dispensing machine for the crystallization screening reduces the required amount of sample to the microgram scale. Absolute configurations of a chiral alkaloid and three chiral pharmaceutical acids were obtained by X-ray crystallography from 210–530 µg of samples by this renovated method.

Background and Objectives: Microbial superantigens have been reported in the blood and synovial fluid of rheumatoid arthritis patients, raising the question of whether the presence of these superantigens could provoke the induction of inflam- matory biomarkers expression or not. The purpose of this study was to examine the Staphylococcus aureus superantigen C on CD18 expression. Materials and Methods: The superantigen C was purified by ultrafiltration. Immunoblotting was performed using a specific antibody. Also, 50 micrograms of superantigens (toxin) were injected intraperitoneally and intra-articularly into separate rat groups. Blood was collected and RNA extracted. Then, the cDNA was synthesized. The expression of CD18 marker was evaluated using RT-real-time PCR, and the results were descriptively analyzed. Results: The results of this study revealed that 50 μg of toxin, injected intra-articularly and intraperitoneally, showed the surplus expression of the marker CD18 in the blood of rats after 20 days. By this method, the expression of the marker CD18 was significantly different between rats that received the superantigen intra-articularly and intraperitoneally (2.10; 2.3 and 3.3 folds) and the controls (P≤ 0.05). Conclusion: The results indicated that the presence of Staphylococcal of superantigen C in the body of rats has enhanced the expression of the CD18 inflammatory marker more than 3 times. This valuable finding is an introduction to further research and could provide new methods to prevent and control inflammatory diseases, including rheumatoid arthritis.

Background Haemophagocytic lymphohistiocytosis (HLH)/macrophage activation syndrome (MAS) is a hyperinflammatory syndrome potentially leading to critical illness. Early treatment reduces mortality but diagnosis requires a high index of suspicion. Highly elevated ferritin levels (HEF) >10,000 μg/L are highly specific for HLH/MAS [1] and should prompt consideration of hyperinflammation. Diagnostic guidelines for HLH, requiring the presence of ≥5/8 criteria [2], and classification criteria for MAS complicating systemic juvenile idiopathic arthritis (sJIA) have been published [3]. Objectives To assess recognition of HLH/MAS in a paediatric population with HEF. Methods This retrospective study was conducted at 11 centres under local service evaluation permissions. Biochemistry databases identified patients ≤16 years with serum ferritin >10,000 μg/L during a 3-year period. Each case was assessed against the 2004 HLH criteria and, for patients with sJIA, the 2016 MAS criteria. Due to limited access to some of the laboratory tests, previously-published modified HLH criteria using a threshold of ≥4/5 (excluding tissue haemophagocytosis, decreased natural killer cell function, increased soluble interleukin-2 receptor) were also applied to all patients [4]. Results 153 patients (55.6% male) were identified. Patient diagnoses included: infections (29.4%), rheumatological (17.0%) and malignancies (17.0%). A diagnosis of HLH/MAS was made by the treating clinical team in 39.9% and considered in a further 16.3%. Using all available data, 30/153 (19.6%) met ≥5/8 criteria and 93.3% of these patients were diagnosed with HLH/MAS by the treating team. 56 (36.6%) met ≥4/5 criteria and 33 (58.9%) of these were diagnosed with HLH/MAS by clinicians. HLH/MAS was not documented as being considered in the differential in 23.2%. Of 23 patients with sJIA, 82.6% met MAS classification criteria and 89.5% of these were diagnosed with MAS by the treating clinicians. Overall mortality was 32.7% (50/153) and was 27.9% (17/61) in patients diagnosed with HLH/MAS during their admission. Conclusion Although HEF is highly specific for HLH/MAS, the diagnosis was only made or considered in just over half of paediatric patients with this laboratory result. Increased awareness of this potentially-lethal condition is likely to lead to earlier treatment and reduced mortality. References [1] Allen CE, et al. 2008;50:1227-35. [2] Henter JI, et al. 2007;48:124-31. [3] Ravelli A, et al. 2016;75:481-9. [4] Davi S, et al. 2014;66:2871-80. Disclosure of Interests Ethan Sen: None declared, Beverley Almeida: None declared, Louise Moran: None declared, Charlene Foley: None declared, Nagla Abdelrahman: None declared, Rosie Close: None declared, Ema-Louise Long: None declared, Joshua Bennett: None declared, Jason Palman: None declared, Catriona Anderson: None declared, Kirsty McLellan: None declared, Samundeeswari Deepak: None declared, Kathy Gallagher: None declared, Peter Bale: None declared, Kamran Mahmood: None declared, Clare Pain: None declared, Flora McErlane: None declared, Athimalaipet Ramanan Consultant for: AbbVie, UCB, Sobi, Eli Lilly, Speakers bureau: Speaker fees/honoraria from Abbvie, SOBI, Eli Lilly and UCB, Speakers bureau: AbbVie, UCB, Sobi, Eli Lilly, Rachel Tattersall: None declared

Background There is variability in the fecal calprotectin (FCP) cut-off level for the prediction of inflammatory bowel disease (IBD) activity and differentiation from irritable bowel disease (IBS). We aimed to assess the status of the optimal FCP cut-off value for the evaluation of IBD. Methods We reviewed the existing literature regarding the optimal FCP cut-off level in patients with IBD for the prediction of disease activity, remission, relapse and differentiation from IBS patients. Results The study reveals the large quantitative differences in FCP cut-off levels in different study populations (from 50 to918 microgram/gram) (table 1). FCP cut-off value for the initial diagnosis of IBD or active disease status ranged from 50 to800 microgram/gram. A cut-off level of 50–250 microgram/gram differentiated patients of IBD from IBS. Cut-off level of FCP for the prediction of remission ranged from 250 to 918 microgram/gram. Cut-off value for the prediction of relapse ranged from 50 to200 microgram/gram. Variability in the cut-off level is due to the use of different test kits and different study populations. Gastrointestinal infections and chronic subclinical intestinal inflammation, may explain the higher cut-off FCP levels in underdeveloped populations. Studies have demonstrated high levels of FCP in patients with intestinal tuberculosis and chronic giardia infection. Conclusions There is a wide variation in FCP cut-off levels in the initial diagnosis of IBD as well as in follow-up post-treatment. The FCP cut-off levels vary from country to country. The FCP test requires validation of the available test kits and finding of appropriate cut-off levels for different study populations.

Background: Normal skeletal development and adult bone maintenance requires normal level of thyroid hormones. Growth arrest, delayed bone maturation, and epiphyseal dysgenesis are fundamental result of hypothyroidism in children. Aim of the work: To detect the effects of hypothyroidism and thyroxin treatment on the postnatal development of lumbar vertebrae in albino rats. Materials and Methods: Sixty adult female rats were randomly subdivided into three equal groups; control group (received distilled water), carbimazole-treated group (received carbimazole in a dose of 6 microgram/gram body weight daily orally from 10th gestational day till the day 21 postnatally then carbimazole administration was continued to pups until the end of 8th week postnatally) and carbimazole+thyroxin treated group (received carbimazole on 10th gestational day in a dose of 6 microgram /gram body weight orally and thyroxin at birth in a dose of 10 microgram/kg/day orally and the administration of both drugs was continued until the day 21 postnatally then the administration was further continued to pups until the end of 8th week postnatally). The offsprings at ages (newborn, two weeks and eight weeks) anesthetized, sacrificed, their lumbar vertebrae were manibulated for histological examination. Morphometric and statistical studies were performed. Results: The lumbar vertebrae markedly affected by hypothyroidism especially the proliferating zone and the collagen component of the matrix. Morphometric results revealed highly-significant decrease in the thickness of the epiphysis in hypothyroid rats. Most of these changes reverted by giving thyroxin. Conclusion: Vertebral column growth is markedly affected by hypothyroidism and thyroxin replacement has a protective role.

Listeria monocytogenes ATCC 19115, Scott A, and UAL500 were evaluated for sensitivity to nisin (0 to 50 microgram/ml) using a direct plating method. Nisin (10 microgram/ml) decreased an initial population of L. monocytogenes (10(9) CFU per ml) by 6- to 7-log cycles. Sensitivity to nisin was enhanced by addition of 2% NaCl or by reduction of the medium pH from 6.5 to 5.5 with either hydrochloric or lactic acid. Mutants resistant to 50 microgram/ml nisin were detected at frequencies of 10-6 to 10-8. Nisin-resistant L. monocytogenes mutants should be expected to arise when nisin is used as an antimicrobial in food systems

International audience; A novel double labelling of glycosaminoglycans (GAG) oligosaccharides by thia-Michael addition and deuterium incorporation at the non-reducing and reducing ends, respectively, was introduced. This was demonstrated to be both compatible with the heparin microgram scale and amenable for mass spectrometry analysis, without impairing enzymatic activities such as heparinase I and sulfatase HSulf-2.

AIM To evaluate the effect of FSLLRY-NH2, a protease-activated receptor 2 (PAR2) inhibitor, on neurocognitive impairment and hippocampal neuronal degeneration in the setting of asphyxial cardiac arrest (ACA)-induced global cerebral ischemia (GCI) in rats. MATERIAL AND METHODS A total of 43 Sprague-Dawley male rats were used. Shams and rats resuscitated from 9 minutes of ACA were randomized to two separate experiments including time course and short-term neurological outcomes. FSLLRY-NH2 (50 microgram [μg] per rat) was administered intranasally at 1 hour postresuscitation. Neurological function and hippocampal neuronal degeneration were evaluated after ACA. RESULTS Significant neurological function decline and hippocampal neuron degeneration were observed in ACA animals as compared with the shams. Treatment with FSLLRY-NH2 significantly improved neurological outcome and reduced the number of degenerating hippocampal neurons after ACA. CONCLUSION Targeting PAR2 may be a novel therapeutic approach in the management of neurological dysfunction after cardiac arrest-associated ischemic injury.

Objectives This study aimed to compare cellular expression of lysyl oxidase-like 1 (LOXL1), a key enzyme in elastin metabolism, of premenopausal women with pelvic organ prolapse (POP) compared with premenopausal controls without POP and postmenopausal women with POP. In addition, we examined whether variation of LOXL1 expression was dependent on biopsy site. Methods A standardized protocol was utilized to obtain vaginal biopsies from 30 women (10 premenopausal POP, 10 postmenopausal POP, and 10 premenopausal non-POP). Expression levels of messenger RNA (mRNA) and protein of LOXL1 were determined using real-time quantitative polymerase chain reactions and enzyme-linked immunosorbant assays. Analysis was performed to determine if there were differences between group or biopsy site. Results Significant differences in LOXL1 mRNA expression were found between patient groups (P = 0.0033). LOXL1 mRNA expression (relative to 18S) was upregulated in the postmenopausal POP group (54.5 ± 14.7) compared with the premenopausal POP group (5.2 ± 14.7, P = 0.0034) and the premenopausal non-POP group (23 ± 18, P = 0.0359). No significant differences in LOXL1 protein expression (nanogram/milliliter per microgram total protein) were seen between groups (premenopausal POP, 3.2 × 10 ± 6.3 × 10; postmenopausal POP, 4.3 × 10 ± 6.3 × 10; premenopausal non-POP, 5.0 × 10 ± 7.7 × 10; P = 0.15). No differences in mRNA expression were seen between sites (P = 0.74), but significant variation was noted in protein expression (P = 0.001). Conclusions Premenopausal and postmenopausal women with POP exhibit differential expression of LOXL1 suggesting different pathways in the pathogenesis of POP. The role of biopsy location on LOXL1 expression requires further investigation.

Background: In the management of first trimester miscarriage, the use of oral misoprostol is beneficial for patients as it offers a more discrete and less invasive route for those women who find vaginal administration unacceptable. In spite of high incidence of side-effects from use of oral misoprostol women still found oral route satisfactory. Methods: This study was a prospective cohort study done at El-Mukala maternal and child hospital and Hadhramout maternal and child university hospital in the period between 1st October 2014 and 30th September 2015. All pregnant women (less than 14 weeks) who were diagnosed as an embryonic pregnancy or missed miscarriage were included in the study. Every patient received single dose of oral misoprostol 600 µg in half full stomach at home. The primary outcome measure was complete miscarriage rate. Results: One-hundred women were included in the study. The mean age of study participants was 26.25±4.08 years, the mean BMI was 27.35±3.6 while the mean parity was 2.6±1.5.Ten cases needed emergency surgical evacuation within the period of first 48 hours. Complete miscarriage had occurred in 75 cases, 65 of them in the first 48 hours. Fifteen cases presented by incomplete miscarriage after waiting for one week. They needed surgical evacuation at the end of 7 days due to still considerable intrauterine contents. Conclusions: In our closed community in El-Mukala, Yemen, the use of oral misoprostol in single dose of 600 µg at home as a method for termination of first-trimester miscarriage was effective (75%, success rate), tolerable regarding side effects, has the advantage of high confidentiality and privacy resulting in good satisfaction.

A banding scheme theory has been proposed to assess the potency/toxicity of biologics and assist with decisions regarding the introduction of new biologic products into existing manufacturing facilities. The current work was conducted to provide a practical example of how this scheme could be applied. Information was identified for representatives from the following four proposed bands: Band A (lethal toxins); Band B (toxins and apoptosis signals); Band C (cytokines and growth factors); and Band D (antibodies, antibody fragments, scaffold molecules, and insulins). The potency/toxicity of the representative substances was confirmed as follows: Band A, low nanogram quantities exert lethal effects; Band B, repeated administration of microgram quantities is tolerated in humans; Band C, endogenous substances and recombinant versions administered to patients in low (interferons), intermediate (growth factors), and high (interleukins) microgram doses, often on a chronic basis; and Band D, endogenous substances present or produced in the body in milligram quantities per day (insulin, collagen) or protein therapeutics administered in milligram quantities per dose (mAbs). This work confirms that substances in Bands A, B, C, and D represent very high, high, medium, and low concern with regard to risk of cross-contamination in manufacturing facilities, thus supporting the proposed banding scheme.

Background: Paravertebral block(PVB),for inguinal hernia repair has been found to be effective than conventional spinal anaesthesia in terms of anaesthetic and analgesic technique , early ambulation and better postoperative pain score. Bupivacaine, an amide local anaesthetic, having a slow onset of action, provides early onset , prolonged duration of analgesia and adequate sedation in case of PVB with an adjuvant dexmedetomidine ,a highly selective alpha2 adrenergic agonist. Aims: To compare the efficacy as well as onset and duration of sensory and motor block ,duration of post operative analgesia ,any complication of the drugs bupivacaine and a combination of bupivacaine with dexmedetomidine in paravertebral block for inguinal hernia repair. Material and Method: 60 patients aged between 18-60 yrs of ASA physical status I and II , randomized in two groups of 30 to receive bupivacaine (0.5%) 1mgkg—1 in control group A and bupivacaine (0.5%) 1mgkg —1combined with dexmedetomidine (1 μgkg—1) in study group B for PVB. Results: Gr A vs Gr B was significant (P