Abnormal Somatosensory Behaviors Associated With a Gain-of-Function Mutation in TRPV3 Channels

Thermosensitive transient receptor potential V3 (TRPV3) is a polymodal receptor implicated in nociceptive, thermoceptive, pruritoceptive, and inflammatory pathways. Reports focused on understanding the role of TRPV3 in thermoception or nociception are not conclusive. Previous studies also show that aberrant hyperactivity of TRPV3 channels results in spontaneous itch and dermatitis-like symptoms, but the resultant behavior is highly dependent on the background of the animal and the skin microbiome. To determine the function of hyperactive TRPV3 channels in somatosensory sensations, we tested different somatosensory behaviors using a genetic mouse model that carries a gain-of-function point mutation G573S in the Trpv3 gene (Trpv3G573S). Here we report that Trpv3G573S mutants show reduced perception of cold, acetone-induced cooling, punctate, and sharp mechanical pain. By contrast, locomotion, noxious heat, touch, and mechanical itch are unaffected in Trpv3G573S mice. We fail to observe any spontaneous itch responses and/or dermatitis in Trpv3G573S mutants under specific pathogen (Staphylococcus aureus)-free conditions. However, we find that the scratching events in response to various pruritogens are dramatically decreased in Trpv3G573S mice in comparison to wild-type littermates. Interestingly, we observe sensory hypoinnervation of the epidermis in Trpv3G573S mutants, which might contribute to the deficits in acute mechanical pain, cool, cold, and itch sensations.

Identification and partial purification of thermally stable peroxidase isoenzymes from seedlings of Vigna sp. (V) landrace Vn

Several soluble peroxidase isoenzymes are expressed in a landrace of Vigna sp. cultivated in the north of Cameroon (landrace called Vn in previous study) during seed germination. There are at least two cathodic peroxidases and eight major anodic peroxidases as shown by their electrophoretic migration at pH 7.4 under native conditions. These isoperoxidases are more expressed in roots than in shoots. They have different thermal stability, so that heat inactivation kinetics of crude peroxidase extracts from roots do not fit the first-order model. The slow and intermediate migrating groups of anodic isoperoxidases retains a substantial activity after ten minutes of incubation at 80°C and 85°C. An anodic isoperoxidase (named A6 in this study) shows in addition to this great thermal stability, a high activity in seedlings and is expressed both in roots and shoots. The combination of those characteristics makes this isoperoxidase a potential candidate for biotechnological applications. Three major anodic isoperoxidases, of which A6 and another thermostable isoperoxidase, were successfully separated from each other by ion exchange chromatography on DEAE-cellulose, after precipitation of total proteins by ice-cold acetone. This offers the prospect of being able to characterize these isoperoxidases individually in future studies.

Polymers Sorption Properties towards Photosynthetic Pigments and Fungicides

In the present work, extraction with a solvent (cold acetone) was used to extract the assimilation pigments from spinach leaves. Then, the sorption capacity of selected plastics granules (polyvinyl chloride—PVC, polypropylene—PP, polyethylene—PE of different densities) was tested for the selective isolation of chlorophylls. Quantification of chlorophylls by HPLC (Zorbax Eclipse XDB-C18 column, the mobile phase: Acetonitrile/methanol/ethyl acetate 6:2:2, v/v) was based on chlorophyll-a content as the most common chlorophyll. The performed experiments prove that PVC containing electronegative chlorine exhibits favorable interactions toward chlorophyll by creating stable molecular complexes. The Fourier Transform Raman Spectroscopy (FT-Raman) and the molecular modeling were used to elucidate the structure of the created complexes. The optimal extraction requirements, the mass of sorbent, water-acetone ratio, time, and the composition of the elution solvent were all established. The optimized extraction conditions ensured a maximum extraction yield of chlorophylls of 98%. The chlorophyll-rich sorbent was re-extracted by acetone, leading to the recovery of 91% of chlorophylls in one step, adding the possibility of its re-use. The proposed effective and ecological method of obtaining the green dye from plants is cheap, simple, and efficient, avoiding organic solvents, utilizing the most widely used synthetic polymers in the world, being products difficult for utilization. The possibility to remove chosen fungicides cyprodinil, chlorothalonil, and thiabendazone from plant extract by PVC was also examined. The described method proposes a new application of synthetic polymers, which meets the criteria of sustainable green chemistry, simultaneously reaching the growing demand for pure natural compounds in the pharmaceutical and food industries.

The antimicrobial and antioxidant property, GC–MS analysis of non-edible oil-seed cakes of neem, madhuca, and simarouba

Background The antimicrobial activity against clinically and agriculturally important microorganisms, antioxidant property and chemical profiling of acetone crude extracts of non-edible oil-seed cakes of neem (Azadirachta indica A. Juss), madhuca (Madhuca longifolia syn. Madhuca indica Gmelin) and simarouba (Simarouba glauca DC) obtained by hot and cold extraction methods were tested in-vitro. Results The hot neem and cold acetone extracts of madhuca and simarouba were inhibitory to Staphylococcus aureus. The enteric pathogens-Enterococcus faecalis and Salmonella enterica were inhibited by both hot and cold extracts of simarouba cake. Fusarium oxysporum and Colletotrichum capsici were sensitive to oil-seed cake extracts of madhuca and simarouba cake extracts followed by neem cake extract. The Aspergillus flavus was highly sensitive to neem followed by madhuca and simarouba extracts. The cyclic voltammetry of all extracts indicated the presence of oxidation peaks at different Epa values suggesting the presence of antioxidant ability. The GC–MS profile revealed the presence of pharmacologically important essential compounds. Conclusion The study revealed the presence of bioactive compounds in non-edible oil-seed cakes that could be exploited for human welfare.

Characterization of Collagen from Sakhalin Taimen Skin as Useful Biomass

Research background. Animal collagen has been widely utilized in foods, cosmetics, and biomedical fields. The non-edible portion, such as fish skins and bones, are generated during cooking processes. Most of them are currently discarded as wastes, although the nutritional values of the skins and bones are high. It needs to utilize the non-edible portion for the reduction of environmental impact, as it may be one of source of environmental pollution. Experimental approach. Collagen was prepared from Sakhalin taimen skins as wastes generated during cooking processes. Next, the color, SDS-polyacrylamide gel electrophoresis, ultraviolet absorption, subunit composition, amino acid composition, denaturation temperature, and attenuated total reflectance-Fourier transform infrared spectroscopy analysis were conducted to explore the properties of the collagen. Lastly, it tried to improve the functional properties of the collagen using chemical modification technique for future applications. Results and conclusions. Cold acetone treatment made it possible to easily remove the fats and pigments from skins. The odorless and pure-white collagen was obtained with high-yield. The α3 chain did not exist in the collagen. Sakhalin taimen skin collagen had rich α-helix and low β-sheet structures. Succinylation caused the secondary structural changes of the collagen molecule. Moreover, succinylation made it possible not only to increase the viscosity of collagen solution and but also to improve the solubility of collagen in the physiological conditions around pH=6. Novelty and scientific contribution. This finding was the first report on the absence of the α3 chain in Salmonid fish skin collagens. The succinylated collagen from Sakhalin taimen skins as useful biomass has potential to utilize in foods, cosmetics, and its related industries.

The effects of crude C. inophyllum seed oil to silica gel mass ratio and number of stages on the isolation of wax

Calophyllum inophyllum is one of the most notable mangrove species that grows a lot in the coastal areas of the Indonesian archipelago. Despite its long lifespan (50 years) and abundant seed oil production, this plant has not been utilized optimally. Wax is reported as a constituent of C. inophyllum seed oil, but the quantity has not been discovered yet. Wax has been commonly used as ingredient in coating, cosmetic, food, and pharmaceutical industries. The aims of this work were to separate wax from C. inophyllum seed oil in high purity and investigate the effects of crude C. inophyllum to silica gel mass ratio and the stages number on wax isolation. Silica gel was employed to adsorb crude C. inophyllum seed oil. Mass ratios of C. inophyllum seed oil to silica gel used in this work were 1:4, 1:2, and 1:1 (g/g). After that, the seed oil adsorbed onto silica gel was extracted by soxhlet extraction with hexane as the solvent. Wax was separated by putting the hexane extract in cold storage at 4 ºC for 24 h. Furthermore, wax obtained was analyzed by High-Temperature Gas Chromatography (HT-GC) and confirmed by Thin-Layer Chromatography (TLC). It was found that wax (purity 93.2 % and yield 0.4 %) was best isolated by employing two-stage of adsorption-extraction, with crude C. inophyllum seed oil to silica gel mass ratio of 1:1 (g/g), followed by crystallization in cold acetone for 24 h. Moreover, wax content in crude C. inophyllum seed oil was 0.43%.

Purification and Characterization of a Protease Inhibitor with Anticancer Potential from Bacillus endophyticus JUPR15

Introduction:Introduction: Protease Inhibitors (PIs) constitute a group of proteins widely distributed among all organisms and their main function includes their ability to inhibit the proteolytic activity. PIs represent an important role in the regulation of various cellular physiological and biological processes, including cell cycle, cell death, differentiation and immune response.Material and Methods:Hence, in our search for novel anticancer compounds, we isolated microorganisms from various environmental sources and screened them for the production of protease inhibitors. Promising isolates were further checked for their protease inhibitory activity by their ability to inhibit the activity of trypsin and chymotrypsin, which were measured spectrophotometrically.Results:The isolate identified as Bacillus endophyticus JUPR15 was found to be promising with higher inhibitory activity than the other isolates. The inhibitor was purified by cold acetone precipitation and column chromatography and further subjected to characterization studies by performing 12 % SDS-PAGE to determine the molecular weight and gelatin-PAGE assay to confirm its inhibitory activity.Conclusion:The isolate exhibited promising anticancer activity on in-vitro Hela and HepG2 cancer cell lines, showing its application potentials.

The capability of the red seaweed Gracilaria vermiculophylla in producing prostaglandins

The red seaweed G. vermiculophylla is rich in polyunsaturated fatty acids with 20 carbon atoms, mainly arachidonic acid (AA) and eicosapentaenoic acid, which are precursors of prostaglandins (PGs). The present study aimed to elucidate the capability of the seaweed in releasing PGs using acetone powder as the crude enzyme. Crude enzyme was prepared using cold acetone. The crude enzyme was incubated with AA at different concentrations (0.1– 4 mg). For determination of PG contents, 5 µL of sample as the test solution corresponding to 0.2 g wet mass of the seaweed was injected into the HPLC. For mass spectrometer analysis, an HPLC system connected with mass spectrometer was used. Results of the study showed that t he released PGs from incubation of acetone powder and AA analyzed by HPLC consisted of PGE 2 , 15-keto-PGE 2 , 15-hydroperoxy-PGE 2 , PGA 2 , and AA while PGs detected by LC-MS were PGF 2α , PGE 2 , 15-keto-PGE 2 , 15-hydroperoxy-PGE 2 , and PGA 2 . The capability of the red algae in producing PGs was affected by available oxygen, aspirin, a cyclooxygenase inhibitor, and AA concentration. The crude enzyme of the red alga (250 mg) was capable to produce 1.63 µg and 1.32 µg of PG 2 and 15-keto-PGE 2 from incubation with 0.25 mg of AA. This method could be the one way to provide PGs in vitro to fulfill demands of PGs in the pharmaceutical industry.