Discovery of a C-S lyase inhibitor for the prevention of human body malodor formation: tannic acid inhibits the thioalcohol production in Staphylococcus hominis.

Human body odor is a result of the bacterial biotransformation of odorless precursor molecules secreted by the underarm sweat glands. In the human axilla, Staphylococcus hominis is the predominant bacterial species responsible for the biotransformation process of the odorless precursor molecule into the malodorous 3M3SH by two enzymes, a dipeptidase and a specific C-S lyase. The current solutions for malodor, such as deodorants and antiperspirants are known to block the apocrine glands or disrupt the skin microbiota. Additionally, these chemicals endanger both the environment and human health, and their long-term use can influence the function of sweat glands. Therefore, there is a need for the development of alternative, environmentally friendly, and natural solutions for the prevention of human body malodor. In this study, a library of secondary metabolites from various plants was screened to inhibit the C-S lyase, which metabolizes the odorless precursor sweat molecules, through molecular docking and molecular dynamics (MD) simulation. In silico studies revealed that tannic acid had the strongest affinity towards C-S lyase and was stably maintained in the binding pocket of the enzyme during 100-ns MD simulation. We found in the in vitro biotransformation assays that 1 mM tannic acid not only exhibited a significant reduction in malodor formation but also had quite low growth inhibition in S. hominis, indicating the minimum inhibitory effect of tannic acid on the skin microflora. This study paved the way for the development of a promising natural C-S lyase inhibitor to eliminate human body odor and can be used as a natural deodorizing molecule after further in vivo analysis.

Determination of temporal changes in hepatic drug-oxidizing capacity using plasma metabolite/caffeine ratios in non-pregnant and pregnant goats.

Caffeine (CF) is a metabolic probe drug used in the determination of the hepatic drug-oxidizing capacity. The aim of this study was to investigate temporal changes in the hepatic drug-oxidizing capacity using plasma metabolite/CF ratios in non-pregnant goats (n = 11) and pregnant goats (n = 23). CF (5 mg/kg, intravenous) was administered in six periods (Period 1-6) with 45 days between two periods. The plasma levels of CF and its metabolites, theophylline (TP), theobromine (TB) and paraxanthine (PX), were determined by HPLC-UV. To evaluate hepatic drug-oxidizing capacity in terms of enzymes that play a role in CF metabolism, the plasma metabolic ratios including TB/CF, PX/CF, TP/CF and TB + PX + TP/CF were determined at 10 h following CF administration. Plasma metabolite/CF ratios were similar between non-pregnant and pregnant goats. However, plasma metabolite/CF ratios in Period 3 (45 days in pregnant goats) were significantly higher than those other periods in both pregnant and non-pregnant goats. The effect of pregnancy may not be observed on drugs that are substrates of enzymes involved in CF metabolism in goats.

The potential of microalgal sources as coating materials: A case study for the development of biocompatible surgical sutures.

Sutures are considered as surgical materials that form excellent surfaces to integrate the postoperative parts of the body. These materials present suitable platforms for potential bacterial penetrations. Therefore, coating these biomedical materials with biocompatible compounds is seen as a potential approach to improve their properties while avoiding adverse effects. The aim of this study was to evaluate Arthrospira platensis, Haematacoccus pluvialis, Chlorella minutissima, Botyrococcus braunii, and Nostoc muscorum as potential surgical suture coating materials. Their crude extracts were absorbed into two different sutures as poly glycolic (90%)-co-lactic acid (10%) (PGLA) and poly dioxanone (PDO); then, their cytotoxic effects and antibacterial activities were examined. Both N. muscorum-coated sutures (PGLA and PDO) and A. platensis-coated (PGLA and PDO) sutures did not induce any toxic effect on L929 mouse fibroblast cells (>70% cell viability). The highest antibacterial activity against Staphylococcus aureus was achieved with N. muscorum-coated PGLA and A. platensis-coated PGLA at 11.18 ± 0.54 mm and 9.52 ± 1.15 mm, respectively. These sutures were examined by mechanical analysis, and found suitable according to ISO 10993-5. In comparison with the commercial antibacterial agent (chlorohexidine), the results proved that N. muscorum extract can be considered as the most promising suture coating material for the human applications.

The heat is on: a simple method to increase genome editing efficiency in plants.

BACKGROUND: Precision genome mutagenesis using CRISPR/Cas has become the standard method to generate mutant plant lines. Several improvements have been made to increase mutagenesis efficiency, either through vector optimisation or the application of heat stress. RESULTS: Here, we present a simplified heat stress assay that can be completed in six days using commonly-available laboratory equipment. We show that three heat shocks (3xHS) efficiently increases indel efficiency of LbCas12a and Cas9, irrespective of the target sequence or the promoter used to express the nuclease. The generated indels are primarily somatic, but for three out of five targets we demonstrate that up to 25% more biallelic mutations are transmitted to the progeny when heat is applied compared to non-heat controls. We also applied our heat treatment to lines containing CRISPR base editors and observed a 22-27% increase in the percentage of C-to-T base editing. Furthermore, we test the effect of 3xHS on generating large deletions and a homologous recombination reporter. Interestingly, we observed no positive effect of 3xHS treatment on either approach using our conditions. CONCLUSIONS: Together, our experiments show that heat treatment is consistently effective at increasing the number of somatic mutations using many CRISPR approaches in plants and in some cases can increase the recovery of mutant progeny.

The rate of oocytes with granular cytoplasm is higher in women with endometrioma in ICSI cycles.

The purpose of this study was to investigate the impact of endometrioma on oocyte morphology and fertility outcome in intracytoplasmic sperm injection (ICSI)cycles. The study material was obtained from 114 ICSI cycles of infertile women aged between 20 and 38 years with ovarian endometriomas and unexplained infertility. In total, 644 mature oocytes were included in the analysis. The rates of specific oocyte morphological abnormalities were similar between the two groups however the central granulation rate was significantly higher in the group with endometrioma (p < .05). Fertilisation rate were not significantly different between the groups (p ≥ .05) however the numbers of metaphase 2 (MII) oocytes and embryos were lower in the endometrioma group (p ≤ .05). Endometrioma was associated with a higher rate of oocytes with granular cytoplasm, despite the fertilisation rate the numbers of the MII oocytes and embryo were affected.IMPACT STATEMENTWhat is already known on this subject? The association between endometrioma and infertility is a well-known condition, but the possible mechanisms of the effects of endometrioma on women's fertility is still debated and controversial. There is limited data on the effect of endometrioma on oocyte morphology. Low oocyte quality and lower fertilisation rates might be the main cause of adverse pregnancy outcomes during in vitro fertilisation/intracytoplasmic sperm injection cycles.What do the results of this study add? Endometrioma was associated with a higher rate of oocytes with granular cytoplasm, and lower metaphase 2 oocytes and embryos.What are the implications of these findings for clinical practice and/or further research? Future studies using further oocyte quality assessment methods and prospective observational studies including live-birth rate should be designed to better understand how endometrioma affects fertility outcomes.

Impaired semen parameters in patients with confirmed SARS-CoV-2 infection: A prospective cohort study.

In this prospective study, we investigated the impact of SARS-CoV-2 infection on semen parameters in a cohort of men who had recently recovered from COVID-19. A total of 24 men who had recently recovered from mild COVID-19 were included in the study. Their semen parameters were normal before COVID-19 according to the World Health Organization 2010 reference values. Semen samples were collected from these participants in the recovery phases of COVID-19. To determine the effect of SARS-CoV-2 infection on semen parameters, the patients' pre-COVID-19 and post-COVID-19 semen analyses were compared. The mean age of the participants was 34.7 ± 6.4 years. The median interval between the positive nasopharyngeal swab test and obtaining semen samples was 111.5 (158) days. There was no significant difference in semen parameters before and after COVID-19 in terms of semen volume (p = .56), sperm concentration (p = .06), and progressive motility (p = .14). Total motility (p = .01) and total motile sperm count (p = .02) decreased significantly after SARS-CoV-2 infection compared to the pre-infection values. This study demonstrated that sperm motility and total motile sperm count were the semen parameters which showed a significant reduction in cases with a history of mild COVID-19.

Hyperbaric oxygen treatment in bilateral orchiopexy and post-circumcision haematoma in a thrombocytopenic patient with Noonan syndrome.

Hyperbaric oxygen treatment (HBOT) can be utilised for necrotising soft tissue infections, clostridial myonecrosis (gas gangrene), crush injuries, acute traumatic ischaemia, delayed wound healing, and compromised skin grafts. Our case was a 17-month-old male patient with Noonan syndrome, idiopathic thrombocytopenic purpura, and bilateral undescended testicles. Haematoma and oedema developed in the scrotum and penis the day after bilateral orchiopexy and circumcision. Ischaemic appearances were observed on the penile and scrotal skin on the second postoperative day. Enoxaparin sodium and fresh frozen plasma were started on the recommendation of haematology. Hyperbaric oxygen treatment was initiated considering the possibility of tissue necrosis. We observed rapid healing within five days. We present this case to emphasise that HBOT may be considered as an additional treatment option in patients with similar conditions. To our knowledge, no similar cases have been reported in the literature.

Chemical Composition and Tyrosinase Inhibitory Activities of Fatty Acids Obtained from Heterotrophic Microalgae, S. limacinum and C. cohnii.

Tyrosinase is the rate-limiting enzyme for melanin production in plant and mammalian cells. Upregulation of this enzyme results in hyperpigmentation disorders. In order to treat pigmentation problems, novel skin whitening compounds are extremely screened. It is found that fatty acids based on their saturation levels either increase or decrease tyrosinase enzyme activity. Thus, fatty acids and their compositions are promising candidates for the treatment of hyperpigmentation or hypopigmentation disorders. Microalgae are rich in both saturated and unsaturated fatty acids, as well. In this study, C. cohnii and S. limacinum fatty acids were evaluated as tyrosinase inhibitor candidates. Mushroom tyrosinase activity studies displayed that both extracts increase tyrosinase enzyme activity dose-dependently. On the other hand, S. limacinum at 200 µg ml-1 concentration almost decreased half of tyrosinase enzyme activity in B16-F10 cells. Besides, it was 3 times more efficient for tyrosinase enzyme activity inhibition and 2 times more effective to decrease melanin synthesis compared to C. cohnii. Considering low toxicity to B16-F10 melanoma and healthy keratinocyte cells (HaCaT), S. limacinum fatty acids could be a suitable source for lipid-based tyrosinase inhibitory functional cosmetics products.

Label-free cellular manipulation and sorting via biocompatible ferrofluids.

We present a simple microfluidic platform that uses biocompatible ferrofluids for the controlled manipulation and rapid separation of both microparticles and live cells. This low-cost platform exploits differences in particle size, shape, and elasticity to achieve rapid and efficient separation. Using microspheres, we demonstrate size-based separation with 99% separation efficiency and sub-10-microm resolution in <45 s. We also show continuous manipulation and shape-based separation of live red blood cells from sickle cells and bacteria. These initial demonstrations reveal the potential of ferromicrofluidics in significantly reducing incubation times and increasing diagnostic sensitivity in cellular assays through rapid separation and delivery of target cells to sensor arrays.

Design of melanogenesis regulatory peptides derived from phycocyanin of the microalgae Spirulina platensis.

Pigmentation issues are common conditions associated with excessive or insufficient production of melanin. Recently peptides are investigated to discover novel melanogenesis regulators as low molecular weight compounds to regulate skin pigmentation. In this study, an internal library of peptides obtained through in silico enzymatic digestion of phycocyanin from microalgae S. platensis was tested to apprehend their anti-melanogenic effects. Seven peptides were investigated for their inhibitory potential against mushroom and B16-F10 murine tyrosinase enzymes. According to the results, P5 (SPSWY) and P7 (AADQRGKDKCARDIGY) were effective in lowering the activity of mushroom and B16-F10 tyrosinases. P5 was the most potent (IC50 value, 12.1 µM) in mushroom which was followed by P2 (MAACLR, 86.9 µM). Although the peptides were particularly powerful in inhibiting monophenolase activity, only moderate inhibition was observed for diphenolase activity in mushroom tyrosinase assay. Apart from tyrosinase inhibition, P2 and P3 (RCLNGRL) were efficient DPPH radical scavengers at low concentrations (IC50 < 200 µM). In the mammalian assay system, P5 and P7 were noticeably effective to decrease tyrosinase enzyme activity with IC50 values of 48.9 and 34.2 µM, respectively. However, although P4 (RYVTYAVF) was a potent mushroom tyrosinase inhibitor, it increased melanin synthesis up to 3-fold in B16-F10 cells. The results indicate that C-terminal tyrosine residue is important for tyrosinase inhibition. This study shows, for the first time, that microalgae proteins can be regarded as sources for melanogenesis regulation.

The effect of medium and light wavelength towards Stichococcus bacillaris fatty acid production and composition.

Introduction of novel species will highlight technical feasibility of microalgae-based biofuels for commercial applications. This paper reports the effect of culture medium and light wavelength on biomass and fatty acid production of S. bacillaris which holds some advantages as short life cycle, easy cultivation, high lipid content, diversity of fatty acids and stability under harsh environmental conditions. The results displayed that, soil extract (SE) greatly enhance growth rate of cultures. Maximum biomass and lipid productivity were achieved in TAP medium as 81 mg/L·day, 19.44 mg/L·day; respectively. Light wavelength didn't significantly change growth kinetics but played a critical role on chlorophyll-a accumulation. C14:0, C16:0 and C18:0 fatty acids were abundant which are suitable for biodiesel conversion. Interestingly, blue and red light increased longer chain fatty acids content. These results indicated that; S. bacillaris holds potential for further development of biodiesel production and feasibility of algal biodiesel for fundamental and applied sciences.

Preparation of electrospun polycaprolactone nanofiber mats loaded with microalgal extracts.

Sustainable, ecological, and biocompatible materials are emerging for the development of novel components for tissue engineering. Microalgae being one of the unique organisms on Earth to provide various novel compounds with certain bioactivities are also a good source for the development of novel tissue scaffold materials. In this study, electrospinning technique was utilized to fabricate nanofibers from polycaprolactone loaded with microalgal extracts obtained from Haematococcus pluvialis (vegetative and carotenoid producing form) and Chlorella vulgaris. The FTIR results showed that, blending microalgae with polycaprolactone give unique bands rooted from microalgae and polycaprolactone structure. The samples were not diversified from each other, however stable bands were observed. SEM analysis revealed a uniform fiber fabrication with an average diameter of 810 ± 55 nm independent from microalgal extracts. MTT assay was done on HUVEC cell lines and results showed that nanofiber mats helped cell proliferation with extended time. Biodegradation resulted with mineral accumulation on the surface of same samples however the fiber degradation was uniform. With slow but stable biodegradation characteristics, microalgal extract loaded nanofiber mats holds great potential to be novel tissue scaffold material.

Photobiological hydrogen production via immobilization: understanding the nature of the immobilization and investigation on various conventional photobioreactors.

Hydrogen photoproduction from microalgae has been an emerging topic for biofuel development. However, low yield for large-scale cultivations seems to be the main challenge. Immobilization seems to be an alternative method for sustainable hydrogen generation. In this study we examined the bead stability, bead diameter and immobilization method in accordance with photobioreactors (PBR). 2.1 mm diameter beads were selected for PBR experiments. CSTR, tubular and panel type PBRs give important results to develop suitable immobilization matrixes and techniques for mass production in scalable PBR systems. In conclusion, we suggest to develop techniques specific for the design and operation characteristic of the PBR for a yield efficient hydrogen generation.

Investigation of in vitro digestibility of dietary microalga Chlorella vulgaris and cyanobacterium Spirulina platensis as a nutritional supplement.

Microalgal proteins are promising sources for functional nutrition and a sustainable candidate for nutraceutical formulations. They also gain importance due to emerging focus on a healthy nutrition and increase in the number of chronic diseases. In this study, dried dietary species of microalga, Chlorella vulgaris, and cyanobacterium Spirulina platensis were hydrolyzed with pancreatin enzyme to obtain protein hydrolysates. The hydrolysis yield of biomass was 55.1 ± 0.1 and 64.8 ± 3.6% for C. vulgaris and S. platensis; respectively. Digestibility, as an indicator for dietary utilization, was also investigated. In vitro protein digestibility (IVPD) values depicted that cell wall structure due to the taxonomical differences affected both hydrolysis and digestibility yield of the crude biomass (p < 0.05). Epithelial cells (Vero) maintained their viability around 70%, even in relatively higher concentrations of hydrolysates in the culture. The protein hydrolysates showed no any antimicrobial activities. This study clearly shows that the conventional protein sources in nutraceutical formulations such as soy, whey, and fish proteins can be replaced by enzymatic hydrolysates of microalgae, which shows elevated digestibility values as a sustainable and reliable source.

Ferrofluid mediated nanocytometry.

We present a low-cost, flow-through nanocytometer that utilizes a colloidal suspension of non-functionalized magnetic nanoparticles for label-free manipulation and separation of microparticles. Our size-based separation is mediated by angular momentum transfer from magnetically excited ferrofluid particles to microparticles. The nanocytometer is capable of rapidly sorting and focusing two or more species, with up to 99% separation efficiency and a throughput of 3 × 10(4) particles/s per mm(2) of channel cross-section. The device is readily scalable and applicable to live cell sorting with biocompatible ferrofluids, offering competitive cytometer performance in a simple and inexpensive package.

The effects of combined application of autogenous fibroblast cell culture and full-tissue skin graft (FTSG) on wound healing and contraction in full-thickness tissue defects.

INTRODUCTION: This study investigates the effects of culture grown fibroblasts on contraction and dermal regeneration when used concurrently with full-thickness skin graft (FTSG) in full-thickness wounds. MATERIALS AND METHODS: Fourteen Sprague-Dawley male rats were divided into two groups. In the first group, wound contraction was evaluated visually. Two full thickness tissue defects were produced on the back of the seven rats. The skin harvested from these areas was prepared as a full-thickness graft and sutured back to their original beds. Just before the last suture, autogenous fibroblast suspension was applied between the graft and the bed in area 1, and area 2 served as control. The surface area of grafts were calculated and compared with "Image J" program. In the second group, contraction and dermal regeneration were evaluated histologically. Three full-thickness tissue defects were produced on the back of seven rats. Area 1 and 2 were prepared as described above and area 3 was left to secondary healing. On the 14th and 30th days, punch biopsies were harvested from the center of the areas 1-3. Preparations were examined under light microscopy. RESULTS: Wound contraction was significantly less in area 1 on day 14 (p<0.01). Histologically neovascularization, fibroblast density and collagen synthesis were more evident in cultured fibroblast applied areas on day 14. However epithelialization did not show any difference between areas both on days 14 and 30. On day 30, area 1 still a higher degree of fibroblast intensity than the other areas but neovascularization and collagen synthesis were not different than the other areas. CONCLUSION: According to the data obtained from the study, cultured fibroblasts, particularly with a dermal support, do not regress when transplanted to a living tissue. They contribute to the wound healing process; reduce the contraction of the wound; and support collagen synthesis and neovascularization.