Isoform Specific Reductions in Na + ,K + -ATPase Catalytic (Α) Subunits in the Nerve of Rats with Streptozotocin-Induced Diabetes

Na + ,K + -ATPase activity in nerve is reduced in rats with streptozotocin-induced diabetes; three different isoforms of the Α (catalytic) subunit of the enzyme are present in nerve. Using western blot to determine subunit isoform polypeptide levels in sciatic nerve, we found a substantial reduction in Α1-isoform polypeptide (88% at 3 weeks, 94% at 8 weeks) after induction of diabetes by streptozotocin. Reductions in Α2 and Α3 polypeptide were smaller and not statistically significant. The reduction in amount of all three isoform polypeptides in the nerve of 3-week diabetic animals was corrected by administration of insulin. Accumulation of Α1 polypeptide at a nerve ligature indicated that rapid transport of that polypeptide in nerve occurs with normal kinetics. The results implicate a specific marked deficit in Α1, much more than Α2 or Α3, catalytic subunit isoform of Na + ,K + -ATPase in the pathogenesis of diabetic neuropathy.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/66384/1/j.1471-4159.1994.63051782.x.pd

Stimulation of liver growth and DNA synthesis by glucosylceramide

The nature of the growth‐stimulating effect of glucosylceramide was studied. Mice were injected intraperitoneally with emulsified glucosylceramide and conduritol B epoxide, an inhibitor of cerebroside glucosidase. Within one or two days, the liver grew 18–24%, as reported. Two enzymes involved in DNA synthesis also increased more than the weight. The total liver activity of thymidine kinase increased 46–73%, and the total activity of ornithine decarboxylase increased as much as 101%. It is suggested that elevated liver levels of glucocerebroside stimulate cell proliferation through a relatively direct mechanism.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/141613/1/lipd0508.pd

Normalization of liver glucosylceramide levels in the "Gaucher" mouse by phosphatidylserine injection

A model of the human genetic disorder, Gaucher disease, can be rapidly generated in mice by the injection of emulsified glucosylceramide and an inhibitor of the lipid's hydrolase, conduritol B epoxide. The liver grows rapidly as it absorbs the load of lipid but the effect disappears as new glucosidase is formed and the load is hydrolyzed. This normalization process is accelerated by treatment with phosphatidylserine, which is a known stimulator of the enzyme. It is possible that injecting the phospholipid into Gaucher patients would have a therapeutic effect since it might help them utilize their residual glucosidase to destroy stored glycolipid.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/27332/1/0000357.pd

Uptake by neuroblastoma cells of glucosylceramide, glucosylceramide glucosidase, its stimulator protein, and phosphatidylserine

Serum-free cultured neuroblastoma cells (clone NIE-115) have been shown to absorb emulsified glucosylceramide, glucosylceramide glucosidase, an activator protein for the enzyme, and phosphatidylserine from a synthetic medium. Uptake of the enzyme was augmented by phosphatidylserine, and vice versa. Uptake of the enzyme-lipid complex was further augmented by the activator protein. It appears likely that the activator forms a complex only with the enzyme-lipid complex, not with the individual components. Two uptake mechanisms for the enzyme seem to be involved, one of which (the complex with activator proteins and acidic lipid) is sensitive to mannosyl phosphate groups. Hydrolysis of absorbed glucosylceramide was slow unless the medium was supplemented with the acidic phospholipid or glucosidase. The most rapid disappearance of stored glycolipid took place when the ternary mixture was added to the cell medium, enzyme + activator protein + phosphatidylserine. These findings may be relevant to enzyme replacement therapy for Gaucher disease.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/26097/1/0000173.pd

Biodegradable PEG-PCL Nanoparticles for Co-delivery of MUC1 Inhibitor and Doxorubicin for the Confinement of Triple-Negative Breast Cancer

Combating triple-negative breast cancer (TNBC) is still a problem, despite the development of numerous drug delivery approaches. Mucin1 (MUC1), a glycoprotein linked to chemo-resistance and progressive malignancy, is unregulated in TNBC. GO-201, a MUC1 peptide inhibitor that impairs MUC1 activity, promotes necrotic cell death by binding to the MUC1-C unit. The current study deals with the synthesis and development of a novel nano-formulation (DM-PEG-PCL NPs) comprising of polyethylene glycol-polycaprolactone (PEG-PCL) polymer loaded with MUC1 inhibitor and an effective anticancer drug, doxorubicin (DOX). The DOX and MUC1 loaded nanoparticles were fully characterized, and their different physicochemical properties, viz. size, shape, surface charge, entrapment efficiencies, release behavior, etc., were determined. With IC(50) values of 5.8 and 2.4 nm on breast cancer cell lines, accordingly, and a combination index (CI) of < 1.0, DM-PEG-PCL NPs displayed enhanced toxicity towards breast cancer cells (MCF-7 and MDA-MB-231) than DOX-PEG-PCL and MUC1i-PEG-PCL nanoparticles. Fluorescence microscopy analysis revealed DOX localization in the nucleus and MUC1 inhibitor in the mitochondria. Further, DM-PEG-PCL NPs treated breast cancer cells showed increased mitochondrial damage with enhancement in caspase-3 expression and reduction in Bcl-2 expression.In vivo evaluation using Ehrlich Ascites Carcinoma bearing mice explicitly stated that DM-PEG-PCL NPs therapy minimized tumor growth relative to control treatment. Further, acute toxicity studies did not reveal any adverse effects on organs and their functions, as no mortalities were observed. The current research reports for the first time the synergistic approach of combination entrapment of a clinical chemotherapeutic (DOX) and an anticancer peptide (MUC1 inhibitor) encased in a diblock PEG-PCL copolymer. Incorporating both DOX and MUC1 inhibitors in PEG-PCL NPs in the designed nanoformulation has provided chances and insights for treating triple-negative breast tumors. Our controlled delivery technology is biodegradable, non-toxic, and anti-multidrug-resistant. In addition, this tailored smart nanoformulation has been particularly effective in the therapy of triple-negative breast cancer. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10924-022-02654-4

Effect of zero tillage basin planting and N nutrition on growth, yield, water productivity and nitrogen use efficiency of late planted broccoli (Brassica oleracea var italica) in North East Hilly Region of India

Our study showed, the zero tillage basin planting with 180 kg N/ha enhanced the marketable yield, water productivity in broccoli (Brassica oleracea L. var italica) and save 71.2% water over flat bed planting with conventional tillage. Study revealed, it is an alternative system for small holder farmers of North East India for growing late planted broccoli under limited water availability

Early Events Associated with Infection of Epstein-Barr Virus Infection of Primary B-Cells

Epstein Barr virus (EBV) is closely associated with the development of a vast number of human cancers. To develop a system for monitoring early cellular and viral events associated with EBV infection a self-recombining BAC containing 172-kb of the Epstein Barr virus genome BAC-EBV designated as MD1 BAC (Chen et al., 2005, J.Virology) was used to introduce an expression cassette of green fluorescent protein (GFP) by homologous recombination, and the resultant BAC clone, BAC-GFP-EBV was transfected into the HEK 293T epithelial cell line. The resulting recombinant GFP EBV was induced to produce progeny virus by chemical inducer from the stable HEK 293T BAC GFP EBV cell line and the virus was used to immortalize human primary B-cell as monitored by green fluorescence and outgrowth of the primary B cells. The infection, B-cell activation and cell proliferation due to GFP EBV was monitored by the expression of the B-cell surface antigens CD5, CD10, CD19, CD23, CD39, CD40 , CD44 and the intercellular proliferation marker Ki-67 using Flow cytometry. The results show a dramatic increase in Ki-67 which continues to increase by 6–7 days post-infection. Likewise, CD40 signals showed a gradual increase, whereas CD23 signals were increased by 6–12 hours, maximally by 3 days and then decreased. Monitoring the viral gene expression pattern showed an early burst of lytic gene expression. This up-regulation of lytic gene expression prior to latent genes during early infection strongly suggests that EBV infects primary B-cell with an initial burst of lytic gene expression and the resulting progeny virus is competent for infecting new primary B-cells. This process may be critical for establishment of latency prior to cellular transformation. The newly infected primary B-cells can be further analyzed for investigating B cell activation due to EBV infection

Multi-criteria multi-facility location in Niwai block, Rajasthan

Rural regions often suffer disproportionately when compared to urban areas in the access to basic healthcare and educational opportunities. The provision of these facilities to the populace has been identified as one of the means of stimulating development in a region. A problem-structuring method with multi-criteria decision analysis was used for the selection of different facilities based on the needs of the rural area under consideration. A facility location model was created and algorithms were developed in order to provide a solution for locating facilities in 45 villages of Niwai block in Tonk district, Rajasthan. Sixteen different facilities were chosen for consideration, each falling into one of five broad groups: healthcare, education, connectivity, agriculture and drinking water. Alternative scenarios for locating facilities were generated and explored, providing a base for the micro-level planning process at the block level in a district