Achiral internucleoside linkages: CH2-CH2-NH and nH-CH2-CH2 linkages

The synthesis of CH₂-CH₂-NH and NH-CH₂-CH₂ internucleoside linkages are described. Antisense oligonucleosides containing these dimer modifications hybridized to the sense sequence. Furthermore incorporation of these backbone modifications enhanced the nuclease resistance of the antisense strand.     

Passage number of cancer cell lines: Importance,intricacies, and way-forward

Cancer cell lines play a crucial role as invaluable models in cancer research, facilitating the examination of cancer progression as well as the advancement of diagnostics and treatments. While they may not perfectly replicate the original tumor, they generally exhibit similar characteristics. Low-passage cancer cell lines are generally preferred due to their closer resemblance to the original tumor, as long-term culturing can alter the genetic and molecular profiles of a cell line thereby highlighting the importance of monitoring the passage number (PN). Variations in proliferation, migration, gene expression, and drug sensitivity can be linked to PN differences. PN can also influence DNA methylation levels, metabolic profiles, and the expression of genes/or proteins in cancer cell lines. When conducting research on cancer cell lines, it is crucial for researchers to carefully select the appropriate PN to maintain consistency and reliability of results. Moreover, to ensure dependability and replicability, scientists ought to actively track the growth, migration, and gene/or protein profiles of cancer cell lines at specific PNs. This approach enables the identification of the most suitable range of PNs for experiments, guaranteeing consistent and precise results. Additionally, such efforts serve to minimize disparities and uphold the integrity of research. In this review, we have laid out recommendations for laboratories to overcome these PN discrepancies when working with cancer cell lines.     

mtDNA diversity in rhesus monkeys reveals overestimates of divergence time and paraphyly with neighboring species

Reconstructions of the human-African great ape phylogeny by using mitochondrial DNA (mtDNA) have been subject to considerable debate. One confounding factor may be the lack of data on intraspecific variation. To test this hypothesis, we examined the effect of intraspecific mtDNA diversity on the phylogenetic reconstruction of another Plio- Pleistocene radiation of higher primates, the fascicularis group of macaque (Macaca) monkey species. Fifteen endonucleases were used to identify 10 haplotypes of 40-47 restriction sites in M. mulatta, which were compared with similar data for the other members of this species group. Interpopulational, intraspecific mtDNA diversity was large (0.5%- 4.5%), and estimates of divergence time and branching order incorporating this variation were substantially different from those based on single representatives of each species. We conclude that intraspecific mtDNA diversity is substantial in at least some primate species. Consequently, without prior information on the extent of genetic diversity within a particular species, intraspecific variation must be assessed and accounted for when reconstructing primate phylogenies. Further, we question the reliability of hominoid mtDNA phylogenies, based as they are on one or a few representatives of each species, in an already depauperate superfamily of primates.      

Growth and analysis of crystal forms of toxic shock syndrome toxin 1

Native toxic shock syndrom toxin 1 (TSST-1) purified from Staphylococcus aurius has been crystallized in four different forms. The highest resolution data (2.05 Å) was collected from orthorhombic crystals belonging to the space group C222₁. The unit cell dimension are a = 108.7 Å, b = 177.5 Å, c = 97.6 Å. Rotation function analysis of this from indicates that there is trimer of toxin molecules in the asymmetric unit with a local 3-fold axis parallel to the crystallographic c axis. Crystals of a double mutant of TSST-1 have been grown which has a single molecule in the asymmetric unit and diffract to 1.9 Å. The space group is P2₁ with unit cell parameters of a = 44.4 Å, b = 34.0 Å, c = 55.2 Å, β = 93.0°. © 1993 Wiley-Liss, Inc.     

Proteomics evaluation of five economical commercial abundant protein depletion kits for enrichment of diseases-specific biomarkers from blood serum

Blood serum is arguably the most analyzed biofluid for disease prediction and diagnosis. Herein, we benchmarked five different serum abundant protein depletion (SAPD) kits with regard to the identification of disease-specific biomarkers in human serum using bottom-up proteomics. As expected, the IgG removal efficiency among the SAPD kits is highly variable, ranging from 70% to 93%. A pairwise comparison of database search results showed a 10%–19% variation in protein identification among the kits. Immunocapturing-based SAPD kits against IgG and albumin outperformed the others in the removal of these two abundant proteins. Conversely, non-antibody-based methods (i.e., kits using ion exchange resins) and kits leveraging a multi-antibody approach were proven to be less efficient in depleting IgG/albumin from samples but led to the highest number of identified peptides. Notably, our results indicate that different cancer biomarkers could be enriched up to 10% depending on the utilized SAPD kit compared with the undepleted sample. Additionally, functional analysis of the bottom-up proteomic results revealed that different SAPD kits enrich distinct disease- and pathway-specific protein sets. Overall, our study emphasizes that a careful selection of the appropriate commercial SAPD kit is crucial for the analysis of disease biomarkers in serum by shotgun proteomics.     

A comparative account of the microbiological characteristics of soils under natural forest,grassland and cropfield from Eastern India

Microbiological and physico-chemical characteristics of tropical forest, grassland and cropfield soils from India were investigated. The study revealed that the conversion of natural forest led to a reduction of soil organic C (26–36%), total N (26–35%), total P (33–44%), microfungal biomass (44–66%) and total microbial biomass C, N and P (25–60%) over a period of 30–50 years. Comparative analysis of microbial activity in terms of basal soil respiration revealed maximum activity in the forest and minimum in the cropfield soil. Analysis of microbial metabolic respiratory activity (qCO₂) indicated relatively greater respiratory loss of CO₂-C per unit microbial biomass in cropfield and grassland than in forest soil. Considering the importance of the microbial component in soil, we conclude that the conversion of the tropical forest to different land uses leads to the loss of biological stability of the soil.     

Efficacy of foliar extracts against pre- and post-harvest diseases of sponge-gourd fruits

S.K. AHMAD AND J.S. PRASAD. 1995. The inhibitory potential of aqueous foliar extracts of Adhatoda vasica, Azadirachta indica, Catharanthes roseus, Datura fistulosa, Lantana camara, Muraya exotica, Ocimum sanctum, Ricinus communis and Strychnos nux-vomica were evaluated against soft rot diseases of sponge-gourd fruits caused by Helminthosporium spiciferum and Fusarium scirpi. Conidial germination of F. scirpi and H. spiciferum were reduced to about 75% when their spores were treated with A. indica, C. roseus, D. fistulosa, L. camara, M. exotica and O. sanctum. Considerable reductions (50%) in mycelial dry weights and colony diameters were recorded in liquid media containing (20%) aqueous extracts of C. roseus, D. fistulosa and M. exotica . Other extracts reduced the mycelial growth but to a lesser extent. Post-infection treatment of sponge-gourd fruits with the extracts of A. indica, L. camara, M. exotica, O. sanctum, D. fistulosa and C. roseus almost fully inhibited the spread of disease. Treatment of fruits with A. indica and C. roseus before fungal infection reduced the spread of disease caused by both the pathogens, whereas M. exotica, O. sanctum and D. fistulosa reduced the spread of soft-rot caused by F. scirpi but not by H. spiciferum .     

Free and total thyroid hormones in humans at extreme altitude

Alterations in circulatory levels of total T₄ (TT₄), total T₃ (TT₃), free T₄ (FT₄), free T₃ (FT₃), thyrotropin (TSH) and T₃ uptake (T₃U) were studied in male and female sea-level residents (SLR) at sea level, in Armed forces personnel staying at high altitude (3750 m) for prolonged duration (acclimatized lowlanders, ALL) and in high-altitude natives (HAN). Identical studies were also performed on male ALL who trekked to an extreme altitude of 5080 m and stayed at an altitude of more than 6300 m for about 6 months. The total as well as free thyroid hormones were found to be significantly higher in ALL and HAN as compared to SLR values. Both male as well as female HAN had higher levels of thyroid hormones. The rise in hormone levels in different ALL ethnic groups drawn from amongst the southern and northern parts of the country was more or less identical. In both HAN and ALL a decline in FT₃ and FT₄ occurred when these subjects trekked at subzero temperatures to extreme altitude of 5080 m but the levels were found to be higher in ALL who stayed at 6300 m for a prolonged duration. Plasma TSH did not show any appreciable change at lower altitudes but was found to be decreased at extreme altitude. The increase in thyroid hormones at high altitude was not due to an increase in hormone binding proteins, since T₃U was found to be higher at high altitudes. A decline in TSH and hormone binding proteins and an increase in the free moiety of the hormones is indicative of a subtle degree of tissue hyperthyroidism which may be playing an important role in combating the extreme cold and hypoxic environment of high altitudes.     

Post-coital agents and menses inducing drugs

The importance of developing of drugs which could be taken post-coitally or used once-a-month in the case of a delay in the onset of the menses is well recognized. The availability of such technology would limit exposure to fertility regulating agents to such occasions where there is coital exposure or possibility of pregnancy.Methods of post-coital contraception used so far include IUD’s inserted post-coitally, estrogens, and combinations of estrogens and gestagens. These are reserved primarily for emergency situations to protect women from unwanted pregnancy resulting from rape or unprotected coitus. Levonorgestrel has shown satisfactory results in terms of contraceptive efficacy and is being further evaluated clinically. A number of problems inherent in the development of post-coital contraception are discussed.Menstrual regulation could be achieved by a number of approaches: (a) block progesterone receptors and interfere with the preparation of the endometrium for implantation; (b) luteolysis leading to decreased progesterone levels and interruption of implantation; and (c) termination of early pregnancy by prostaglandins. A number of progesterone antagonists have been evaluated. One of the compounds, RU38486 is being evaluated clinically for termination of very early pregnancy.Deglycosylated derivatives of human chorionic gonadotropin have been shown to antagonize the action of human chorionic gonadotropin and interfere with established pregnancy in rats. Appropriate methods of delivery, immunogenicity and alternate methods for production of human chorionic gonadotropin need to be considered before evaluation of the derivatives for clinical use. In vitro and invivo models need to be developed for evaluation of the teratogenicity and embryotoxicity of post-coital and menses inducing agents.There are a number of gaps in the knowledge of the processes regulating implantation which should be investigated in rodents and in different non-human primate species.