The Pharmacology of Botulinum Toxin Type A

The aim of this chapter is to structure current information clarifying the most disputable issues of botulinum neurotoxin type A (BoNT/A) pharmacology after systemic (botulism) impact and local medical application. Botulinum neurotoxin (BoNT) pharmacological features evaluated open ways to study factors affecting its biological activity: to extend/shorten its effect duration, to increase/decrease BoNT sensitivity in specific patient populations. The chapter presents unique molecular mechanisms underlying BoNT/A pharmacokinetics and pharmacodynamics: entering the body, distribution, receptor binding, translocation, mediator release suppression, zinc metabolism as well as factors affecting body sensitivity to BoNT at each of those stages. The specific biological effects of BoNT/A, which may underlie its analgesic, anticancer and anti-inflammatory effects, are described. Botulinum neurotoxin pharmacokinetics and pharmacodynamics features discussed herein represent significant clinical relevance since they determine botulinum treatment safety and effectiveness. And also they open ways to develop both BoNT-based therapies and anti-botulinic agents.

Stage specific comparative transcriptomic analysis to reveal gene networks regulating iron and zinc content in pearl millet [Pennisetum glaucum (L.) R. Br.]

Pearl millet is an important staple food crop of poor people and excels all other cereals due to its unique features of resilience to adverse climatic conditions. It is rich in micronutrients like iron and zinc and amenable for focused breeding for these micronutrients along with high yield. Hence, this is a key to alleviate malnutrition and ensure nutritional security. This study was conducted to identify and validate candidate genes governing grain iron and zinc content enabling the desired modifications in the genotypes. Transcriptome sequencing using ION S5 Next Generation Sequencer generated 43.5 million sequence reads resulting in 83,721 transcripts with N50 of 597 bp and 84.35% of transcripts matched with the pearl millet genome assembly. The genotypes having high iron and zinc showed differential gene expression during different stages. Of which, 155 were up-regulated and 251 were down-regulated while during flowering stage and milking stage 349 and 378 transcripts were differentially expressed, respectively. Gene annotation and GO term showed the presence of transcripts involved in metabolic activities associated with uptake and transport of iron and zinc. Information generated will help in gaining insights into iron and zinc metabolism and develop genotypes with high yield, grain iron and zinc content.

Vitamin D nemesis of COVID-19

The surge in the spread of the corona virus disease (COVID-19) pandemic alerted us to opt for the preventive medicine, as prevention is always better than cure. Apart from wearing mask, frequent hand washing and social distancing, strengthening our immune response plays a pivotal role in preventing infections. Vitamin D not only aids in calcium and phosphate homeostasis but also acts as an immunomodulator; the deficiency of which is linked with various respiratory and systemic infections. Hence we took up this review to study the effect of vitamin D in corona illness. Vitamin D exerts the expression of pro-inflammatory cytokines, hinders zinc metabolism, lowers Interleukin 6 levels and thereby inhibits cytokine storm in covid patients. Studies have proved that the covid patients have vitamin D deficiency and its supplementation improves the disease severity as well as the length of hospital stay. To conclude, Vitamin D supplementation can protect as well as halt the progression of corona virus disease. Further trials are needed to set the therapeutic levels in various stages of corona illness.

PSX-B-25 Milk productivity and zinc species analysis in cattle depending on the level of lead accumulation

The aim was to study the milk productivity and zinc species in dairy Holstein cow blood serum depending on the level of lead accumulation. Fat and protein content in milk was assessed by the FIL-IDF procedure. The major zinc species in dairy cow blood serum were tested using a hybrid high-performance liquid chromatography-inductively coupled plasma-mass spectrometry technique. The lead content in the blood serum of all animals was in the intervals of the physiological standard. Animals with low-normal values (Pb concentration up to the 25th percentile) and high-normal values (Pb concentration above the 75th percentile) were taken to form comparison groups. Animals with high-normal values of lead had a statistically significant decrease of milk yield by 21.8% and daily production of fat by 16.4%. There was a tendency towards a decrease in the yield of protein and its percentage. It was found that the zinc content in all studied fractions (α 2-macro-globulin tetramer, α 2-macro-globulin dimer, albumin, citrates, total content) was statistically significantly lower (more than 4.8 times) in the group with high-normal values of lead. The change in the redistribution of zinc-binding ligands was found depending on the level of lead in the blood. The relative content of the α2-macro-globulin tetramer and α2-macro-globulin dimer was significantly higher by 7% and by 13% in the group with high-normal values of lead. There was the decrease in the level of animal productivity, the metal-ligand forms of zinc underwent changes despite the absence of a significant difference in the total level of lead in the blood serum. This indicates a high potential for the use of speciation analysis in the laboratory diagnosis and allows predicting and correcting zinc metabolism. The studies were performed in accordance with the research plan for 2019–2021 of the Federal Research Center of Biological Systems and Agrotechnologies RAS No. 0526-2019-0001.

Comparative Transcriptome Analysis in Monocyte-Derived Macrophages of Asymptomatic GBA Mutation Carriers and Patients with GBA-Associated Parkinson’s Disease

Mutations of the GBA gene, encoding for lysosomal enzyme glucocerebrosidase (GCase), are the greatest genetic risk factor for Parkinson’s disease (PD) with frequency between 5% and 20% across the world. N370S and L444P are the two most common mutations in the GBA gene. PD carriers of severe mutation L444P in the GBA gene is characterized by the earlier age at onset compared to N370S. Not every carrier of GBA mutations develop PD during one’s lifetime. In the current study we aimed to find common gene expression signatures in PD associated with mutation in the GBA gene (GBA-PD) using RNA-seq. We compared transcriptome of monocyte-derived macrophages of 5 patients with GBA-PD (4 L444P/N, 1 N370S/N) and 4 asymptomatic GBA mutation carriers (GBA-carriers) (3 L444P/N, 1 N370S/N) and 4 controls. We also conducted comparative transcriptome analysis for L444P/N only GBA-PD patients and GBA-carriers. Revealed deregulated genes in GBA-PD independently of GBA mutations (L444P or N370S) were involved in immune response, neuronal function. We found upregulated pathway associated with zinc metabolism in L444P/N GBA-PD patients. The potential important role of DUSP1 in the pathogenesis of GBA-PD was suggested.

ANALYSIS OF VITAMIN D WITH INTRACTABLE DISEASES- A REVIEW.

Vitamin D is a fat-soluble, steroid hormone responsible for the phospho calcium mechanism. Mainly vitamin D encompasses a group of steroid compounds, namely the VitD2 (ergocalciferol) and the VitD3(cholecalciferol). It is an important requirement for normal functioning of the skeletal muscles, organs and normal metabolism. Providing Vitamin D supplementation to people with Diabetes (Type 1 and 2), Covid-19 positive and people with Thyroid problems showed improved glycaemic controls and regularized blood sugar levels, increase in immune response and regulating the Thyroid levels in the body. Vitamin D also adds to the innate immune system and also plays a major role in zinc metabolism which becomes a major reason for decreasing the symptoms of COVID-19 in the human body. Keywords: Vitamin D, Type 2 Diabetes Mellitus, Type 1 Diabetes Mellitus, Covid-19, Diabetic Neuropathy, Diabetic Nephropathy, Thyroid, Hypovitaminosis.

Stage Specific Comparative Transcriptomic Analysis To Reveal Gene Networks Regulating Iron And Zinc Content In Pearl Millet [Pennisetum glaucum (L.) R. Br.]

Pearl millet is an important staple food crop of poor people which is rich in micronutrients like iron and zinc and amenable for focused breeding for these micronutrients along with high yield. Transcriptome sequencing using ION S5 Next Generation Sequencer generated 43.5 million sequence reads resulting in 83,721 transcripts with N50 of 597 bp and 84.35% of transcripts matched with the pearl millet genome assembly. The genotypes having high Fe and Zn showed differential gene expression during different stages. Of which, 155 were up-regulated and 251were down-regulated while during flowering stage and milking stage 349 and 378 transcripts were differentially expressed, respectively. Gene annotation and GO term showed the presence of transcripts involved in metabolic activities associated with uptake and transport of iron and zinc. In the present study, the 83,721 transcripts were also examined for identification of SSRs. A total of 4,327 SSRs were identified with dominance of tri-nucleotide SSRs in comparison to di-nucleotide SSRs. These EST-SSRs can be used in molecular breeding, genetic diversity analysis and determination of heterozygosity of the allelic loci. Information generated will help in gaining insights into iron and zinc metabolism and develop genotypes with high yield, grain iron and zinc content.

Effect of bis-glycinate bound zinc or zinc sulfate on zinc metabolism in growing lambs

To assess the efficacy of bis-glycinate bound Zn, 36 crossbred wethers (34 ± 2 kg) were sorted by body weight into three groups and stagger started on a Zn-deficient diet (18 mg Zn/kg dry matter [DM]; 22.5% neutral detergent fiber [NDF]) for 45 d prior to a 15-d metabolism period (10 d adaptation and 5 d collection). On day 46, lambs were randomly assigned to dietary treatments (four lambs treatment−1group−1): no supplemental Zn (CON) or 15 mg supplemental Zn/kg DM (ZINC) as Zn sulfate (ZS) or bis-glycinate (GLY; Plexomin Zn, Phytobiotics). Blood was collected from all lambs on days 1, 44, 56, and 61. Liver, jejunum, and longissimus dorsi samples were collected after euthanasia on day 61. Gene expression was determined via quantitative real-time polymerase chain reaction. Data were analyzed using ProcMixed of SAS (experimental unit = lamb; fixed effects = treatment, group, and breed) and contrast statements assessed the effects of supplemental Zn concentration (ZINC vs. CON) and source (GLY vs. ZS). After 15 d of Zn supplementation, plasma Zn concentrations were greater for ZINC vs. CON and GLY vs. ZS (P ≤ 0.01); tissue Zn concentrations were unaffected (P ≥ 0.27). Liver Cu concentrations were lesser for ZINC vs. CON (P = 0.03). Longissimus dorsi Mn concentrations were greater for ZINC vs. CON (P = 0.05) and tended to be lesser for GLY vs. ZS (P = 0.09). Digestibility of DM, organic matter (OM), and NDF was lesser for ZINC vs. CON (P ≤ 0.05); acid detergent fiber digestibility tended to be greater for GLY vs. ZS (P = 0.06). Nitrogen retention (g/d) tended to be greater for GLY vs. ZS (P = 0.10), and N apparent absorption was lesser for ZINC vs. CON (P = 0.02). Zinc intake, fecal output, retention, and apparent absorption were greater for ZINC vs. CON (P ≤ 0.01). Apparent absorption of Zn was −5.1%, 12.8%, and 15.0% for CON, ZS, and GLY, respectively. Nitrogen and Zn retention and apparent absorption were not correlated for CON (P ≥ 0.14) but were positively correlated for ZINC (retention: P = 0.02, r = 0.52; apparent absorption: P < 0.01, r = 0.73). Intestinal expression of Zn transporter ZIP4 was lesser for ZINC vs. CON (P = 0.02). Liver expression of metallothionein-1 (MT1) tended to be greater for GLY vs. ZS (P = 0.07). Although Zn apparent absorption did not differ between sources (P = 0.71), differences in post-absorptive metabolism may be responsible for greater plasma Zn concentrations and liver MT1 expression for GLY-supplemented lambs, suggesting improved bioavailability of GLY relative to ZS.

Effect of Zincum metallicum and LPS treatments on femoral bone density in pregnant mice

Background: The simulated bacterial infection is a classical experimental model, by the injection of purified gram negative bacteria wall lipopolysaccharide (LPS) which produces effects similar to real infection, but without presence of the microorganism. The LPS is a stress factor as during the pregnancy and delivery, and pilot experiments have shown that ultra-high dilutions (UHD) may have a beneficial role in reducing the LPS stress. Zincum metallicum (Zm) has a potential to influence several physiological aspects of a living organism. It was chosen by a team of researchers from GIRI, to explore the effect of Zm on various experimental model (Zincum project). The optical density is a radiographic method for the measurement of the amount of hard bone tissue, and is an accessible, simple and non-invasive method. It is hypothesized that UHD Zm may influence the Zinc metabolism, therefore, bone tissues were collected to evaluate the zinc concentration.

Nitrogen, Iron and Zinc Acquisition: Key Nutrients to Aspergillus fumigatus Virulence

Aspergillus fumigatus is a ubiquitous soil decomposer and an opportunistic pathogen that is characterized by its large metabolic machinery for acquiring nutrients from media. Lately, an ever-increasing number of genes involved in fungal nutrition has been associated with its virulence. Of these, nitrogen, iron, and zinc metabolism-related genes are particularly noteworthy, since 78% of them have a direct implication in virulence. In this review, we describe the sensing, uptake and regulation process of the acquisition of these nutrients, the connections between pathways and the virulence-implicated genes. Nevertheless, only 40% of the genes mentioned in this review have been assayed for roles in virulence, leaving a wide field of knowledge that remains uncertain and might offer new therapeutic and diagnostic targets.