Enzyme Responsive Vaginal Microbicide Gels Containing Maraviroc and Tenofovir Microspheres Designed for Acid Phosphatase-Triggered Release for Pre-Exposure Prophylaxis of HIV-1: A Comparative Analysis of a Bigel and Thermosensitive Gel

The challenges encountered with conventional microbicide gels has necessitated the quest for alternative options. This study aimed to formulate and evaluate a bigel and thermosensitive gel, designed to combat the challenges of leakage and short-residence time in the vagina. Ionic-gelation technique was used to formulate maraviroc and tenofovir microspheres. The microspheres were incorporated into a thermosensitive gel and bigel, then evaluated. Enzyme degradation assay was used to assess the effect of the acid phosphatase enzyme on the release profile of maraviroc and tenofovir microspheres. HIV efficacy and cytotoxicity of the microspheres were assessed using HIV-1-BaL virus strain and HeLa cell lines, respectively. Maraviroc and tenofovir release kinetics followed zero-order and Higuchi model kinetics. However, under the influence of the enzyme, maraviroc release was governed by first-order model, while tenofovir followed a super case II transport-mechanism. The altered mode of release and drug transport mechanism suggests a triggered release. The assay of the microspheres suspension on the HeLa cells did not show signs of cytotoxicity. The thermosensitive gel and bigel elicited a progressive decline in HIV infectivity, until at concentrations of 1 μg/mL and 0.1 μg/mL, respectively. The candidate vaginal gels have the potential for a triggered release by the acid phosphatase enzyme present in the seminal fluid, thus, serving as a strategic point to prevent HIV transmission.

Soil microbial dynamics and enzyme activities as influenced by biofertilizers and split application of vermicompost in rhizosphere of wheat (Triticum aestivum. L.)

Aim: A field experiment was conducted during rabi season to determine the effect of biofertilizers and split application of vermicompost on biological properties (microbial biomass carbon and nitrogen, microbial populations and enzyme activities) in rhizosphere of wheat. Methodology: The experiment was laid out in factorial randomized block design with three replications consisting of twenty treatment combinations. Soil samples were collected from the plots at 0-15 cm depth after harvest of wheat crop and soil biological properties analyzed using standard analytical procedure. Results: The experiment results indicated that among biofertilizers treatments, seed inoculation with Azotobacter + PSB + KMB + ZnSB (B5) resulted in a significant higher soil microbial biomass carbon, microbial biomass nitrogen, population of bacteria, fungi and actinomycetes, dehydrogenase activity and acid phosphatase enzyme activity in comparison to control. Similarly, application of vermicompost as 50 % VC at sowing + 50 % VC at tillering (V3) were obtained improved microbial biomass carbon and nitrogen, microbial population, dehydrogenase activity and acid phosphatase enzyme activity while remaining at par with 75 % VC at sowing + 25 % VC at tillering (V4) proved superior in comparison to rest of the treatments due to continuous supply of nutrients throughout the crop cycle. Grain and straw yield of wheat also increased due to the application of biofertilizers and vermicompost over the control. Interpretation: Biofertilizers (Azotobacter, PSB, KMB and ZnSB) and split application of vermicompost enhanced the soil microbial population and enzymatic activities which sustained the soil health for better wheat production.

Malathion Biodegradation by L. casei (NRRL1922) and L. acidophilus (NRRL 23431) in Fermented Skimmed Milk

The aim of this study was to investigate and trace the biodegradation products of the pesticide malathion in a comparative manner by two different lactobacilli strains; L. casei (NRRL1922) and L. acidophilus (NRRL 23431). The two strains were cultivated separately into skimmed milk supplemented with 5 ng/ml malathion. After incubation under the appropriate conditions, randomized samples were taken at intervals 24, 48, 72 and 120 hours along with control samples and analyzed for the presence of malathion and its degradation products by the GC-MS spectrometry; As well as, analyzed to record the level of phosphatase enzyme which suggested to be involved in the biodegradation process. The results showed a high ability of the two tested strains to degrade malathion with a superiority of L. acidophilus (NRRL 23431) over L. casei (NRRL 1922). The level of phosphatase enzyme was elevated in both strains in the presence of malathion and decreased gradually upon the depletion of malathion from the sample, which reflects the role of the phosphatase enzyme in the biodegradation process.

Integrated carbon-nitrogen-phosphorus cycling for sustainable agriculture – a knowledge gap and investigation of the role of phosphatase enzymes

<p>Phosphorus is closely linked to other nutrient cycles, notably carbon and nitrogen, therefore, to understand potential risks to food production models are required that simulate integrated nutrient cycling over long timescales. The soil-plant system model N14CP meets these requirements and simulates both semi-natural and agricultural environments. N14CP has been validated both spatially and temporally across a range of long-term agricultural experimental sites comparing soil C, N and P, and crop yields, and in most instances performs well. However, under experimental conditions where N is applied in the absence of P, the model indicates exhaustion of P reserves and a decline in yields that is not observed at these sites, highlighting a gap in the model process representation. Potential sources of this ‘missing P’ such as enhanced atmospheric deposition, weathering and flexible plant stoichiometries were explored yet cannot account for this deficit. We hypothesise that access of organic P through other mechanisms not fully represented within the model, such as phosphatase enzymes, could be part of this explanation.</p><p>In order to test this, we conducted a meta-analysis of phosphatase enzyme activity in agricultural settings, comparing response to P sufficient and deficient conditions. Results suggest phosphatase enzyme activity is higher in P deficient conditions compared to inorganic P addition, yet lower compared to organic P addition. Meta-regression analysis indicates magnitude of P addition and pH of substrate are significant factors influencing enzyme response. However, due to numerous additional processes and adaption strategies in response to P deficiency and the difficulty isolating the role of phosphatase enzymes it is not possible to determine the degree to which this mechanism alone accounts for the missing P. We discuss the continuing need for additional empirical evidence to understand the cycling of organic P, and the development of models to include these processes to inform sustainable land management and ensure long-term food security.</p>

Acid Phosphatase and Zinc Tests Are Effective for Semen Examination and Identification to Prove Intercourse

The presence of spermatozoa in vagina is a definite sign of sexual intercourse. However, sometimes microscopic examination does not find spermatozoa or reveals a false negative result. This is influenced by many factors, including the absence of ejaculate in the vaginal canal. In addition, there are other factors such as oligo/azoospermia, vasectomy, degeneration of sperm due to time, incorrect sampling, and improper storage. Therefore, examination of the other components of the ejaculate, ie. the enzyme acid phosphatase, choline and spermin, is important. Compared with spermatozoa, the enzyme phosphatase, choline and spermin have lower evidentiary value because these three components are less specific. However, the level of phosphatase enzyme found in the vagina is much lower than phosphatase enzyme that comes from prostate gland. In this study, as many as 192 samples in the form of patches with sperm/semen stains were tested with acid phosphatase test and zinc test through direct and indirect examination. In the first method, washing was carried out on day 1, day 7, and month 3, and testing was carried out after each washing. In the second method, washing was carried out simultaneously and testing was carried out on day 1, day 7 and month 3. Washing was carried out using 7 types of detergent for each 4 patch samples, and washing using water as control. The results showed very low sensitivity (0.186) and very high specificity (100%). This showed that both tests had high specificity values. Acid phosphatase test specifically showed the presence of the enzyme phosphatase, while zinc test specifically showed the presence of zinc in semen. This phosphohydrolase-phosphatase enzyme is easily degraded due to external factors, including temperature, humidity, and chemicals, ie. the element SDS (Sodium Dodecyl Sulfate) that has the ability to cut enzymes. The weakness of the acid phosphatase test is that this enzyme is easily degraded, either partially or completely, due to external factors, such as temperature, humidity, heat, and the presence of chemicals.

Smartphone Based Highly Sensitive Visualized Detection of Acid Phosphatase Enzyme Activity

With the rapid development of point-of-care (POC) technologies, the improvement of sensitive method featured with fast analysis and affordable devices has become an emerging requirement for the practical application. In...