Water Bath Research Articles

A novel system with robust compatibility and stability for detecting Sugarcane yellow leaf virus based on CRISPR-Cas12a.

Sugarcane yellow leaf virus (SCYLV) can reduce sugarcane productivity. A novel detection system based on reverse transcription-multienzyme isothermal rapid amplification (RT-MIRA) combined with CRISPR-Cas12a, named RT-MIRA-CRISPR-Cas12a, was developed. This innovative approach employs crude leaf extract directly as the reaction template, streamlining the extraction process for simplicity and speed. Combining RT-MIRA and CRISPR-Cas12a in one reaction tube increases the ease of operation while reducing the risk of aerosol contamination. In addition, it exhibits sensitivity equivalent to qPCR, boasting a lower detection limit of 25 copies. Remarkably, the entire process, from sample extraction to reaction completion, requires only 52-57 minutes, just a thermostat water bath. The result can be observed and judged by the naked eye.IMPORTANCESugarcane yellow leaf disease (SCYLD) is an important viral disease that affects sugarcane yield. There is an urgent need for rapid, sensitive, and stable detection methods. The reverse transcription-multienzyme isothermal rapid amplification combined with CRISPR-Cas12a (RT-MIRA-CRISPR-Cas12a) method established in this study has good specificity and high sensitivity. In addition, the system showed good compatibility and stability with the crude leaf extract, as shown by the fact that the crude extract of the positive sample could still be stably detected after 1 week when placed at 4°C. RT-MIRA-CRISPR-Cas12a, reverse transcription polymerase chain reaction (RT-PCR), and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) were used to detect SCYLV on 33 sugarcane leaf samples collected from the field, and it was found that the three methods reached consistent conclusions. This Cas12a-based detection method proves highly suitable for the rapid on-site detection of the SCYLV.

Formulasi Sabun Cair Berbahan Baku Virgin Coconut Oil VCO dengan Penambahan Ekstrak Daun Pepaya Carica Papaya L. untuk Peningkatan Aktivitas Anti Bakteri Staphylococcus Epidermidis ATCC 12238

Acne is a chronic inflammatory disease caused by Staphylococcus epidermidis. Papaya leaves (Carica papaya L.) contain antibacterial substances, namely alkaloids carpaine, flavonoids, tocophenols. Of this study was to determine the physical quality of liquid soap, increasing antibacterial activity. Papaya leaves were extracted with 96% ethanol solvent, then evaporated with a water bath. Antibacterial activity test using the disc diffusion method in concentrations of 5%, 15% and 25% and liquid soap will be made F0 0%, F1 5%, F2 10%, and F3 25% against Staphylococcus epidermidis ATCC 12238. Amoxicillin tab as a positive control and distilled water as a negative control. Bacterial activity appears a clear zone around the disc paper. And see the physical quality test according to the requirements. The results of the bacterial test data are made in table form, and analyzed using the SPSS method Formula with the addition of papaya leaf extract can be made and according to the physical quality conditions of liquid soap. The results of the 25% concentration extract test are an inhibition zone of 20.8 mm, the highest liquid soap formula is an inhibition zone of 23.8 mm in inhibiting the growth of Staphylococcus epidermidis ATCC 12238. Liquid soap preparations and papaya leaf extract have antibacterial activity against Staphylococcus epidermidis ATCC 12238 bacteria and according to the physical quality requirements of liquid soap.

In Vitro Kinetic and Thermodynamic Staining Study of Three Energy Drinks on the Restorative Charisma Composite

Abstract Aim: The research utilized charisma composite resin, a dental restorative material known for its esthetic properties, with the chemical formula 2,2-bis[p(2-hydroxy-3-methacryloxy propoxyphenyl)] propane. The primary objective of the study was to examine the overall color change (ΔE*ab) of the charisma resin after it was subjected to three distinct energy drink solutions: Monster Energy Ultra-Sunrise, Wild Tiger, and Red Bull. Further we examined the effects of temperature and time on the staining interaction and overall color difference ΔE*ab on the surface of charisma restorative resin generated by three energy drink. Materials and Methods: A microfilled Charisma Classic composite resin was used in presented comparative study selected. The prepared 144 composite disk samples (subdivided n=48) with random allocation method were used for the three energy drinks to determine the effect of temperature and time over the course of 90 days. A metallic circulator mold was prepared. Every set of 48 specimens in the energy drink was heated in a water bath for 1, 7, 30, 60, and 90 days at four different temperatures This investigation spanned various time intervals, ranging from 1 to 90 days, and encompassed four temperature levels between 283 and 310 K. One-way ANOVA was used to evaluate the mean data for color change between the energy drinks, and Tukey’s post hoc test was used for multiple comparisons, with a significance level of P < 0.05. Results: The study revealed that the rate of staining was most pronounced in the case of Wild Tiger, while it was comparatively lower for Monster Energy Ultra-Sunrise. These findings were established through an examination of the kinetic and thermodynamic behavior of surface color changes in micro-hybrid composites treated with the three energy drink solutions. Conclusions: The observed patterns aligned with the pseudo second-order model. It was observed that the Red Bull drink displayed a negative activation energy, resulting in a slower color change rate with increase in temperature. In contrast, endothermic, spontaneous, and regular staining activity was demonstrated throughout time by Monster Energy Ultra-Sunrise and Wild Tiger.

Evaluation of Cleaning Methods for Lithium Disilicate Ceramic Post Try-In Paste Application: An SEM Analysis.

This in vitro study assessed the efficacy of three cleaning methods on lithium disilicate ceramic after the application of different try-in pastes through SEM analysis. Ten rectangular specimens of IPS e.max CAD were prepared using a diamond disc, crystallized, etched with 5% hydrofluoric acid, and subjected to three try-in pastes-Calibra ©, Variolink (V), RelyX Veneer®-and three cleaning techniques-air-water spray (RD), ultrasonic bath in distilled water for five minutes (ULT/W), and ultrasonic bath in distilled alcohol for five minutes (ULT/A). A control specimen was also included. After one-minute paste application and subsequent cleaning method application, SEM evaluation was conducted. The results indicate that RD was as effective as CTRL in removing remnants from R-RD, V-ULT/W and V-ULT/A samples, but ineffective for all Calibra paste-contaminated specimens. In conclusion, the optimal removal of try-in paste residues from lithium disilicate restorations is paste-dependent; however, ultrasonic baths with distilled water or alcohol proved effective for most pastes tested.

Formation mechanism and stability of egg white fluid gels under ultrahigh-pressure homogenization pretreatment and synergistic heating effect

The thermal sterilization process of protein beverages inevitably leads to the formation of insoluble thermal aggregates, greatly reducing the texture and taste of protein beverages. In this study, homogenized egg white (HEW) was obtained by ultrahigh-high-pressure (UHP) homogenization pretreatment of egg white (EW), and then a special egg white fluid gel (EWFG) was prepared by water bath heating. The results showed that the optimal conditions for preparing EWFG were three cycles at 20 MPa homogenizing pressure and heating in a water bath at 72℃ for 10 min. Under these conditions, the EWFG was a milky-white homogeneous liquid with an average particle size of about 560 nm. Measurements of the physicochemical properties of HEW and EWFG showed that the UHP homogenization treatment reduced the viscosity of HEW, decreased the particle size of protein aggregates, and increased the zeta potential, which altered the interactions of proteins during the subsequent heating process and facilitated the formation of homogeneous and dispersed EWFG. EWFG showed good stability at pH 6–10 and in low-concentration saline and medium-concentration sucrose solutions. The EWFG obtained by the present treatment is more suitable for factory-scale production and has great potential for protein beverage applications.

Assessing fecal contamination from human and environmental sources using Escherichia coli as an indicator in rural eastern Ethiopian households - a study from the EXCAM project.

Enteric pathogens are a leading causes of diarrheal deaths in low- and middle-income countries. The Exposure Assessment of Campylobacter Infections in Rural Ethiopia (EXCAM) project, aims to identify potential sources of bacteria in the genus Campylobacter and, more generally, fecal contamination of infants during the first 1.5 years of life using Escherichia coli as indicator. A total of 1,310 samples (i.e., hand rinses from the infant, sibling and mother, drinking and bathing water, food and fomite provided to or touched by the infants, areola swabs, breast milk and soil) were collected from 76 households between May 2021 and June 2022. Samples were assigned to two groups by infant age: TP1 (time point 1), infants between 4 and 8 months of age, and TP2, infants between 11 and 15 months of age. Fluorometric and semi-selective colorimetric approaches were used to quantify E. coli in the field samples. Overall, E. coli was ubiquitous within selected households (56.8% across the study). E. coli was more frequently detected than average (>53%) with high concentration (>2-log CFU) in soil (g) and per pair of hand, while the opposite trend (<33%; <1.5-log CFU) was observed in food provided to the infants (g or ml), per areola, and breast milk (ml; P<0.01). E. coli was frequently detected in fomites touched by the infants, drinking and bathing water (>51%), but at low concentration (<1.5-log CFU). Correlation analysis between E. coli concentration in different sample types suggested that the mother's hands might play a key role in the transmission of E. coli between the environment (i.e., soil, bathing water and fomites) and other family members (i.e., infant and sibling; P<0.04; r2>0.3). Using E. coli as surrogate, our study identified mother (hands and areola) as reservoirs likely to be involved in frequent transmission of fecal contaminants to infants within rural Ethiopian households.

Total arsenic and arsenic species in selected seafoods: Analysis using high-performance liquid chromatography- inductively coupled plasma mass spectrometry and health risk assessment

The objective of the present study was to determine the inorganic and organic arsenic species and assess the actual risk of selected seafood produced in Sri Lanka in terms of inorganic arsenic species. A total of 80 samples, comprising 75 seafood samples and 5 freshwater samples, including mollusks, echinoderms, and crustaceans, were analyzed for their total arsenic (TA) levels and arsenic species. TA levels in seafood were determined using Inductively Coupled Plasma Mass Spectrometry (ICP-MS) after microwave digestion. Arsenic species, including arsenobetaine (AsB), arsenite (AsIII), arsenate (AsV), monomethylarsonic acid (MMA), and dimethylarsinic acid (DMA), were selectively separated and quantified using High-Performance Liquid Chromatography (HPLC) coupled to ICP-MS, following water bath extraction with 30 mmol/L nitric acid. The TA levels in seafood ranged from 0.036 to 11.686 mg kg-1. The spike recovery values for the five arsenic species lay within 80 % to 120 %, and the chromatographic recovery percentages ranged from 85.5 % to 123.6 %. In most samples, 100 % AsB was detected. Sea cucumber and both marine and freshwater giant river prawns showed AsB percentages of the total arsenicals at 83.2 %, 39.7 %, and 21.3 %, respectively. DMA was found in sea cucumber and marine giant river prawn at levels of 0.065 mg kg-1 and 0.048 mg kg-1, respectively, while MMA was not detected in any samples. AsIII was found in marine water giant river prawn and freshwater giant river prawn at levels of 0.010 mg kg-1 and 0.044 mg kg-1, respectively. AsV was only found in marine giant river prawn at a concentration of 0.193 mg kg-1. No other samples contained detectable levels of inorganic arsenicals. The Target Hazard Quotient (THQ) values for marine and freshwater giant river prawn were 0.90 and 0.2, respectively, indicating no significant health risks since the THQ values were <1. Therefore, our study suggests that consuming the studied seafood does not pose any health risks in terms of arsenic.

Efficient photocatalytic hydrogen evolution of Z-scheme BiVO4/ZnIn2S4 heterostructure driven by visible light

Solar hydrogen production photocatalysis technology faces a significant challenge in designing and synthesizing efficient photocatalysts to achieve rapid migration of photo-induced charge carriers. This study introduces a method for preparing a BiVO4/ZnIn2S4 (BZS) Z-scheme photocatalyst through cation-exchange and low-temperature water bath synthesis. This system accelerates the separation and migration of photo-induced charges to precisely adjust the redox centers at the BiVO4/ZnIn2S4 heterogeneous interface, enhancing the redox capabilities of both holes and electrons. Consequently, the BiVO4/ZnIn2S4 heterojunction demonstrates outstanding photocatalytic performance. It reveals that BZS-0.3 (with 0.3 mmol BiVO4) achieves a photocurrent density of 0.42 mA cm−2 at 1.70 V vs. NHE (8.4 times that of BiVO4) and exhibits superior hydrogen evolution performance under visible light, reaching 2243 mmol g-1h−1 (19 times that of BiVO4). Cyclability tests confirm the good stability of BZS-0.3, offering a promising photocatalyst for efficient hydrogen production and presenting a new strategy for developing exceptional photocatalytic materials.

Effects of Hyperbaric (Non-Thermal) Sanitization and the Method of Extracting Pomegranate Juice on Its Antioxidant and Antihypertensive Properties.

Pomegranate (Punica granatum L.) is considered a functional food due to its polyphenol content that benefits the body. The type of processing the fruit undergoes is important, as this also influences the concentrations of these compounds. The pomegranate juice was extracted by two methods: manual extraction using a manual juicer through heat treatment in a water bath (Man-P), and extraction through mechanical pressing using Good Nature X-1 equipment and hyperbaric sanitization (Mech-Hyp). Bromatological analyses showed significant differences (p ≤ 0.05) between the two treatments. When subjected to hyperbaric sanitization, the juice showed higher concentrations of moisture, soluble solids, protein, and carbohydrates. In an antioxidant analysis, the ABTS radical showed no significant difference in the treatments, with 96.99% inhibition. For the DPPH radical, the sample with the highest inhibition was Man-P with 98.48%. The determination of phenols showed that there was a higher concentration in juice that underwent pasteurization (104.566 mg GAE/mL). However, the Mech-Hyp treatment exhibited a minor concentration of phenols with 85.70 mg GAE/mL. FTIR spectra revealed that the functional groups were mainly associated with carbohydrates. Regarding ACE inhibition, it was observed that the Man-P and Mech-Hyp juices showed greater inhibition of enzyme in hypertensive patients compared to normotensive patients. This activity can be attributed to the mechanisms of action of antioxidant compounds. Both extraction methods manual and mechanical pressing resulted in increased antioxidant and antihypertensive activity. The antioxidant compounds accompanied by adequate sanitation were decisive in an antimicrobial analysis, since no pathogenic microorganisms were observed in the juices.

Characterization of a fly ash-based hybrid well cement under different temperature curing conditions for natural gas hydrate drilling

This paper presents a study on the temperature effect (3°C, 10°C, and 50°C, in water bath) and activator effect on the characterization of a fly ash-based hybrid well cement, with a large fraction of fly ash (approximately 70 %), which has potential for natural gas hydrate drilling due to its low heat release. The mechanical strength, micro/nanostructure characteristics were investigated through various tests, including x-ray diffraction (XRD), thermogravimetry analysis (TGA-DTG), nuclear magnetic resonance (MAS NMR), and scanning electron microscope (SEM), to reveal the hydration mechanism of the fly ash-based hybrid well cement. The results indicate that the low curing temperatures (3°C, 10°C) inhibited both the hydration of the well cement and the pozzolanic reaction of fly ash in the cement system. However, the incorporation of solid Na2SO4 significantly contributed to the pozzolanic reaction during the hydration process by consuming more calcium hydroxide, promoting more ettringite formation and reducing the apparent activation energy of the blend cement (from 53.06 kJ·mol−1 to 41.23 kJ·mol−1), resulting in better compressive strength. MAS NMR results showed that the addition of solid Na2SO4 facilitated the substitution of Al with Si atoms, leading to the formation of more C-A-S-H and C-(N)-A-S-H gels in the system with a denser microstructure. Overall, the fly ash-based hybrid well cement can be developed as a cement system for natural gas hydrate drilling in the ocean. This cement not only exhibits low heat release but also shows improved strength development, with a 72-hour compressive strength of 8.05 MPa when cured at 10 °C.

Design and synthesis of in situ self-assembled PDI/Bi2MoO6 complexes: Promoting singlet oxygen selective generation and visible light photocatalytic purification performance

Solar-driven photocatalytic degradation of antibiotics has emerged as an eco-friendly approach to treating wastewater. And singlet oxygen is a selective and highly active reactive oxygen species. Bi2MoO6 microsphere photocatalyst modified by narrow band gap PDI organic supramolecular was prepared by water bath heating method to achieve the selective generation of non-free radical. It focused on the effects of different acid-induced assembled and different light sources on the photocatalytic performance of PDI/Bi2MoO6 (PDI/BMO) photocatalysts and explored their ability to remove various pollutants. The results showed that PDI/BMO composites self-assembled with nitric acid (1% PMN) could effectively purify a variety of pollutants under visible light irradiation, including CIP, TC, LVF and AO7. In addition, 1% PMN showed excellent photocatalytic performance in actual water, and even have fair degradation efficiency under red light irradiation. Holes and singlet oxygen were confirmed to be the main reactive oxygen species during CIP degradation by capture experiments and EPR, and the steady-state concentration of singlet oxygen was quantified. Three degradation pathways of CIP under the action of non-radicals were clarified, and the biotoxicity of CIP treated products was evaluated by bacteriostatic experiments. This work could provide an ecologically friendly method for wastewater treatment.

Acute care of cyclic vomiting syndrome and cannabinoid hyperemesis syndrome in the home and emergency department for: Special supplement/proceedings of 3rd international symposium.

Cyclic vomiting syndrome (CVS) and cannabinoid hyperemesis syndrome (CHS) are both characterized by episodic, acute transitions from asymptomatic states to highly symptomatic states of nausea, repetitive vomiting, and often severe abdominal pain. Patients with CVS and CHS face significant challenges to abort or mitigate episodes at home and often require emergency department (ED)-based care. This paper reviews the current treatment approach to abort acute CVS and CHS episodes at home and in ED settings. Multiple pharmacologic and nonpharmacologic interventions have been demonstrated to potentially abort CVS or CHS episodes. Systemic pharmacologic agents often used as abortive therapy include triptans, antiemetics, anxiolytics, NK-1 receptor antagonists, antipsychotics, sedatives in general, and various analgesic / anti-inflammatory medications. Nonsystemic, nonpharmacologic approaches include reducing external stimuli (quiet room, dim lights, etc.), and hot water bathing or the application of topical capsaicin cream. More research is needed to develop evidence-based, individualized abortive treatment plans, as well as to determine whether the abortive treatment for CVS requires a fundamentally different approach than for CHS. When home-based approaches fail, all patients with CVS or CHS should receive nonjudgmental, informed, and compassionate care in the ED to abort their episode. Patients with more severe forms of CVS/CHS who require more frequent ED utilization should develop care plans with their ED to assure predictable and effective treatment.

Effects of Pectinase on Bacterial Succession during Hemp Retting

Pectinase accelerates hemp retting. An elevated temperature during the enzymatic action is favorable for bacterial colonization. Industrial hemp (Cannabis sativa L.) bast fiber was retted in a 40 °C water bath under four different conditions: water retting, pectinase retting, bacterial retting, and bacterial retting with the presence of pectinase. Bacterial communities were sampled from the retting liquid of each condition at the beginning of retting and on days 1, 3, and 5. The bacterial successions were identified by 16S rRNA gene metagenomic sequencing. The results showed that Bacillaceae dominated the hemp retting conditions containing 1% (m/v) pectinase, suggesting that pectinase can manipulate the bacterial community succession by changing the nutrients available to bacteria through the breakdown of pectin. Micromorphological analysis also observed the degradation of a gum-like substance and the aggregation of bacteria with the addition of pectinase.

Standardized Sperm Cryofreezing Technique for Caspian Sea Sturgeons (Huso huso, Acipenser persicus, Acipenser stellatus, Acipenser nudiventris)

Sturgeons, considered `living fossils,` face extinction due to various factors, according to the International Union for Conservation of Nature's Red List. The present study describes a uniform sperm cryopreservation method for the endangered Caspian Sea sturgeon species, which are beluga (Huso huso), Persian (Acipenser persicus), stellate (Acipenser stellatus), and ship (Acipenser nudiventris) sturgeons for both artificial reproduction and ex-situ conservation attempts. For this aim, sperm samples were diluted 1:1 in extender composed of 118 mM Tris buffer (pH=8) and 23.4 mM sucrose-based extender containing 15% DMSO and equilibrated for one hour at 4°C. Then, sperm samples were placed into 0.50-ml straws and multi-stage cooling was applied to freeze with the help of a programmable freezer with an accuracy of ±0.1C/min, and finally stored in liquid nitrogen (-196C) tank. Thawing was performed at 40°C water baths for 25 s. The results indicated that storage for two years resulted in a 24.7% fertilization rate for the stellate sturgeon, while the Persian sturgeon had a remarkable rate of 58.2% after five years storage.

Sustainable route for silica preparation from silica fume

ABSTRACT Silica fume (SF) is a major voluminous and bulky by-product of the ferrosilicon industry, and its disposal poses a significant environmental concern. To address this issue, a sustainable approach was employed to transform SF into silica powder using a precipitation method. The process involved calcination, acid precipitation, aging, and drying, utilising industrial by-products such as silica fume and calcium oxide. Various parameters, including hydrochloric acid concentration, water bath temperature, aging pH, aging temperature, and aging time, were systematically investigated to optimise the properties of the resulting silica product. The physical and chemical attributes of the processed silica were thoroughly examined using techniques such as X-ray diffraction (XRD), X-ray fluorescence (XRF), scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), laser particle size analysis, and dibutyl phthalate (DBP) absorption tests. Under optimal conditions (hydrochloric acid concentration of 20%, water bath temperature of 90℃, aging pH 3-4, aging temperature of 90℃, and aging time of 8 hours), the resulting silica product achieved a purity of 98.5866%, a DBP absorption value of 2.85 mL/g, and a particle size of 6.07 µm, meeting national industry standards. This environmentally benign and cost-efficient synthesis route offers a practical solution for large-scale production.

Performing a physiologically relevant test for cladribine tablets

Introduction. The introduction of devices – analogues of GIS (hereinafter–- Gastro-intestinal simulator) is one of the current ways to develop in-vitro assessment of the quality of solid dosage forms. Testing on a physiologically relevant test device (hereinafter referred to as PRT) makes it possible to predict pharmacokinetic profiles due to more relevant conditions, including the use of biorelevant dissolution media, physiological volumes of the gastrointestinal tract, as well as transit between them.Aim. Conduct a study of cladribine tablets on a physiologically relevant tester in order to predict the behavior of the drug in the human gastrointestinal tract.Materials and methods. The objects of the study are "Mavenclad®, tablets, 10 mg" (series 2200754, expiration date until 04.2025, NERPHARMA, S.r.L., Italy) and "Cladribine, tablets, 10 mg" of domestic production with valid expiration date. During the study, the reagents necessary for the preparation of biorelevant dissolution media and quantitative determination by HPLC. Physiologically relevant test were carried out using an apparatus of our own production, consisting of a DT-6 dissolution tester (ERWEKA GmbH, Germany), a water bath equipped with a Thermomix WB-4 heating element (B. Braun, Germany), and peristaltic pumps (Kamoer, China). The quantitative content of released cladribine was assessed using a highly efficient liquid chromatograph "Khromatek-Kristall HPLC 2014" (ZAO SKB "Khromatek", Russia) using a validated method at a wavelength of 252 nm, analysis time – 7 min, column – Grace HPLC Column Platinum C18-EPS, 250 × 4.6 mm, 5 mm (Grace, USA), temperature – 35 °C, elution mode – isocratic (A : B 80 : 20), mobile phase A – 0.1 % H3PO4 solution, phase B – acetonitrile.Results and discussion. Profiles were obtained to assess the dynamics and degree of release of the studied drugs in various parts of the human gastrointestinal tract. Despite the expected degradation of cladribine in an acidic environment (pH1.2), under physiologically relevant conditions, the drug reached the third section (small intestine model) without degradation. Complete release of cladribine from the test and reference dosage forms was observed. Also, in the future, based on the data obtained, it is possible to predict pharmacokinetic profiles using physiologically based pharmacokinetic modeling approaches.Conclusion. A PSF study was conducted for the drugs "Mavenclad®, tablets, 10 mg" and "Cladribine, tablets, 10 mg". Quantitative determination was carried out by HPLC-UV method. The test results showed complete release of both drugs and reaching the intestinal tract, indicating the absence of degradation of cladribine in the region simulating the stomach.

Inhibiting methanogenesis by targeting thermodynamics and enzymatic reactions in mixed cultures of rumen microbes in vitro.

Mitigation of enteric methane (CH4) emissions from ruminant livestock represents an opportunity to improve the sustainability, productivity, and profitability of beef and dairy production. Ruminal methanogenesis can be mitigated via two primary strategies: (1) alternative electron acceptors and (2) enzymatic inhibition of methanogenic pathways. The former utilizes the thermodynamic favorability of certain reactions such as nitrate/nitrite reduction to ammonia (NH3) while the latter targets specific enzymes using structural analogs of CH4 and methanogenic cofactors such as bromochloromethane (BCM). In this study, we investigated the effects of four additives and their combinations on CH4 production by rumen microbes in batch culture. Sodium nitrate (NaNO3), sodium sulfate (Na2SO4), and 3-nitro-1-propionate (3NPA) were included as thermodynamic inhibitors, whereas BCM was included as a enzymatic inhibitor. Individual additives were evaluated at three levels of inclusion in experiments 1 and 2. Highest level of each additive was used to determine the combined effect of NaNO3 + Na2SO4 (NS), NS + 3NPA (NSP), and NSP + BCM (NSPB) in experiments 3 and 4. Experimental diets were high, medium, and low forage diets (HF, MF, and LF, respectively) and consisted of alfalfa hay and a concentrate mix formulated to obtain the following forage to concentrate ratios: 70:30, 50:50, and 30:70, respectively. Diets with additives were placed in fermentation culture bottles and incubated in a water bath (39°C) for 6, 12, or 24h. Microbial DNA was extracted for 16S rRNA and ITS gene amplicon sequencing. In experiments 1 and 2, CH4 concentrations in control cultures decreased in the order of LF, MF, and HF diets, whereas in experiments 3 and 4, CH4 was highest in MF diet followed by HF and LF diets. Culture pH and NH3 in the control decreased in the order of HF, MF, to LF as expected. NaNO3 decreased (p < 0.001) CH4 and butyrate and increased acetate and propionate (p < 0.03 and 0.003, respectively). Cultures receiving NaNO3 had an enrichment of microorganisms capable of nitrate and nitrite reduction. 3NPA also decreased CH4 at 6h with no further decrease at 24 h (p < 0.001). BCM significantly inhibited methanogenesis regardless of inclusion levels as well as in the presence of the thermodynamic inhibitors (p < 0.001) while enriching succinate producers and assimilators as well as propionate producers (p adj < 0.05). However, individual inclusion of BCM decreased total short chain fatty acid (SCFA) concentrations (p < 0.002). Inhibition of methanogenesis with BCM individually and in combination with the other additives increased gaseous H2 concentrations (p < 0.001 individually and 0.028 in combination) while decreasing acetate to propionate ratio (p < 0.001). Only the cultures treated with BCM in combination with other additives significantly (padj < 0.05) decreased the abundance of Methanobrevibacter expressed as log fold change. Overall, the combination of thermodynamic and enzymatic inhibitors presented a promising effect on ruminal fermentation in-vitro, inhibiting methanogenesis while optimizing the other fermentation parameters such as pH, NH3, and SCFAs. Here, we provide a proof of concept that the combination of an electron acceptor and a methane analog may be exploited to improve microbial efficiency via methanogenesis inhibition.

Fe(Ⅲ)-doped ZnIn2S4-modified graphene oxide in-situ for the degradation of tetracycline hydrochloride in simulated sunlight

To improve the efficiency of photocatalytic reactions, it is important to understand how to achieve fast directional migration and separation of photogenerated carriers at the interface. Here, Fe(III)-doped ZnIn2S4 (ZIS)-coated graphene oxide (GO) composite (GZF) was in-situ prepared via the low-temperature water bath method. Ultrathin Fe(III)-doped ZIS nanosheets were uniformly grown upright on the GO surface. The prepared samples were studied via, such as, X-ray diffraction, scanning electron microscopy combined with energy-dispersive spectroscopy, transmission electron microscopy, ultraviolet spectroscopy, N2 adsorption–desorption measurements, X-ray photoelectron spectroscopy, and electron spin resonance. Compared with ZnIn2S4 (ZIS)-coated graphene oxide (GZ), the GZF exhibit a 2.26-fold increase in the photocatalytic degradation performance of tetracycline hydrochloride (TCH) in simulated sunlight. Through density functional theory calculations, found that Fe(III) doping introduces new energy levels into ZIS, which effectively narrow its bandgap and thus enhance its light absorption efficiency. It was found that h+, ·O2−, and 1O2 were the major reactive oxygen species involved in the photocatalysis, and the effect of pH on the catalytic degradation of TCH was examined. Electron spin resonance and density functional theory results indicate that Fe(III) acts as an electron trap in the ZIS structure to promote the separation of photogenerated electron-hole pairs, which in turn enhances the photocatalytic efficiency of GZF-25.

A Paper-based Loop-Mediated Isothermal Amplification (LAMP) Assay for Highly Pathogenic Avian Influenza.

Avian influenza outbreaks have had significant economic and public health consequences worldwide. Therefore, prompt, reliable, and cost-effective diagnostic devices are crucial for scrutinizing and confining highly pathogenic avian influenza viruses (HPAIVs). Our study introduced and evaluated a novel paper-based loop-mediated isothermal amplification (LAMP) test for diagnosing the H5 subtype of the avian influenza virus (AIV). We meticulously designed and screened LAMP primers targeting the H5-haemagglutinin (H5-HA) gene of AIV and fine-tuned the paper-based detection assay for best performance. The paper-based LAMP assay demonstrated a detection limit of 500 copies per reaction (25 copies/µL). This inexpensive, user-friendly point-of-need diagnostic tool holds great promise, especially in resource-limited settings. It only requires a water bath for incubation and enables visual detection of results without special equipment. Overall, the paper-based LAMP assay provides a promising method for rapidly and reliably detecting the H5 subtype of AIV, contributing to improved surveillance and early intervention strategies.

Can hydrogen prevent stress fractures in athletes?

As a result of the ability of hydrogen rich water to delay muscle fatigue, and considering that tired striated muscles do not sufficiently protect the bone from mechanical stress, the hypothetical mechanism according to which the consumption of the respective drink by athletes could reduce the incidence of stress fractures is discussed. The application of patches with transdermal release of molecular hydrogen in the areas where stress fractures occur frequently could strengthen this preventive action, due to the local anti-inflammatory and regenerative action established experimentally on the senescent bone, to which could be added the stimulation of osteogenesis, the inhibition of bone resorption and the prevention of fractures due to the antioxidant action. The hypothesis can be tested experimentally using the rabbit tibia as a model, or statistical studies can be done on the incidence of stress fractures in athletes who consume hydrogen rich water. Even baths in water saturated with molecular hydrogen can prevent stress fractures, because they reduce the perception of muscle fever, and the plasters could also prevent the complications of this type of fractures.