Tailor-made recombinant prokaryotic lectins for characterisation of glycoproteins.

Development of biosimilars is costly, where glycan analysis is a significant constraint on time and money. This paper provides an in-depth characterisation of several novel recombinant prokaryotic lectins (RPLs), developed through directed evolution, displaying specific binding activities to α-mannose, ß-galactose, fucose and sialic acid residues, tested against major biosimilar targets. The binding characterisation of all lectins was performed employing the principles of bio-layer interferometry (BLI), with help of the streptavidin-coated sensor with the biotinylated lectins. The binding activity of the RPLs and the specificity to a broad range of glycoproteins and glycoconjugates were evaluated and compared to those of equivalent plant-derived lectins. While exhibiting better or similar specificity, RPLs displayed significantly better binding in all cases. The binding mechanisms are explained with particular focus on the role hydrogen bonding plays in the change of specificity for a galactose specific lectin. Furthermore, different sets of RPLs and their plant equivalents were assayed against the different glycoprotein targets to evaluate the analytical parameters of the lectin-glycoprotein interaction. The obtained LoDs reached by the RPLs were lower than those of their plant counterparts apart from one, exhibiting RPL:PL LoD ratios of 0.8, 2.5, 14.2 and 380 for the sets of lectins specific to fucose, α-mannose, ß-galactose and sialic acid, respectively. Such enhancement in analytical parameters of RPLs shows their applicability in protein purification and as bioanalytical tools for glycan analysis and biosensor development.

Total chemical synthesis of lassomycin and lassomycin-amide.

Herein we report a practical synthetic route to the lasso peptide lassomycin () and C-terminal variant lassomycin-amide (). The biological evaluation of peptides and against Mycobacterium tuberculosis revealed that neither had any activity against this bacterium. This lack of biological activity has led us to propose that naturally occurring lassomycin may actually exhibit a standard lasso peptide threaded conformation rather than the previously reported unthreaded structure.

High through-put and highly sensitive liquid chromatography-tandem mass spectrometry separations of essential amino acids using active flow technology chromatography columns.

A preliminary investigation was undertaken to assess the performance of a new chromatography column technology in applications involving liquid chromatography coupled to mass spectrometry. The new column design allows mobile phase and solute to be extracted from the radial central region of the column, which reduces the solvent load to the mass spectrometer and improves separation efficiency. Effectively the column functions as a 'wall-less' column. The advantages of this design is that the analysis through-put can be increased by a factor of five, while at the same time there is a reduction in baseline noise, which results in an increase in the signal to noise response by up to 10-fold in comparison to standard columns with the same internal diameter and approaching 66-fold in comparison to standard columns with the same virtual internal diameter.

Condom knowledge, history of use, and attitudes among chemically addicted populations.

The need for behavioral change of risky sexual practices has been of the highest priority since the onset of the AIDS epidemic. The major focus of education for safe sex has been emphasis on condom use. We surveyed 124 individuals applying to treatment for various chemical dependencies and 60 individuals applying for non-chemical-dependency medical treatment on various aspects of condom knowledge, history of use, and attitudes. Respondents reported that AIDS has motivated them to increase their use of condoms, however, only 13.9% always use them. Education is needed in the areas of increasing protection. Along with the use of a condom, the need for a reservoir tip and the risks associated with multiple sex partners should be stressed.

PIP: This report contains the findings of a survey designed to investigate the knowledge of condoms, history of use, and attitudes among chemically attitudes among chemically addicted populations. The study took place in the Division of Alcohol and Drug Dependence of the SUNY Health Science Center at Brooklyn, Department of Psychiatry, Kings Country Addictive Disease Hospitals. Researchers interviewed 124 individuals seeking treatment for chemical dependency, as well as 60 individuals applying for non-chemical dependency treatment to serve as controls. 25% of those interviewed reported intravenous injection as their primary route of drug administration, 25% reported smoking crack of sniffing cocaine, 17.4% cited alcohol abuse. The remaining 32.6% made up the control group. In a few instances, drug users scored better than the controls on condom knowledge, but overall, the survey found no significant differences in the level of knowledge about AIDS and condoms use among the drug, alcohol, and control groups. As expected, the survey found that those individuals who have a history of condom use scored higher on the knowledge quiz than those with less experience. Very few individuals in any of the groups mentioned monogamy as a strategy for risk reduction, or mentioned multiple sex partners as high-risk activity. While sensitivity and embarrassment did not play a significant role in condom use frequency, 26.1% of those interviewed agreed with the statement "If my partner doesn't mention using a condom neither will I." 32% of males and 57.1% of females reported having has a sex partner refuse to use a condom. These findings, the report explains, suggest the need to address sexually risky behavior within the chemically addicted populations.

AIDS-related risk behaviors among substance abusers.

Ninety-two substance abusers were surveyed about AIDS-related risk behaviors. The questionnaire administered elicited information on demographics, sexual practices, substance abuse, and risk behaviors involving the use of needles. Results indicated an appropriate base of prevention knowledge and patients' efforts to modify vulnerability to HIV exposure. Nevertheless, patients continue to engage in behaviors resulting in HIV exposure and transmission.

A behavioral measure of AIDS information seeking by drug and alcohol inpatients.

A nonreactive, observational research method was used to measure AIDS information-seeking behavior of patients on four drug and alcohol inpatient units at a large public hospital in New York City. Results showed only 23 inquiries from 271 male and female patients over a six-week interval. Possible explanations and implications of these results are discussed.