Dry Bath Research Articles

Ultra-low temperature flexible supercapacitor based on hierarchically structured pristine polypyrrole membranes

• A self-standing PPy electrode (PPy-N) with a hierarchical structure was constructed. • PPy-N possessed superior electrochemical properties and low-temperature flexibility. • PPy-N assembled supercapacitors stably powered electronics in a dry ice bath (<-50 ℃) even cyclic bending. Low-temperature flexible supercapacitors (LFSCs) are urgently needed because most supercapacitors become rigid and prone to damage at extremely low temperatures, such as in the winter of the northern atmosphere, at high altitude, and in space. Although extensive investigations on anti-freezing electrolytes have significantly promoted the development of low-temperature resistant supercapacitors, a usable LFSC remains a major challenge because of the inferior capacitance and flexibility of the existing electrodes at low-temperature. Herein, a supercapacitor prototype that remained flexible and energy storage functional at −50 ℃ was demonstrated, thanks to a hierarchically-structured self-standing pristine polypyrrole membrane (PPy-N) that showed high flexibility and electrochemical performances at ultra-low temperature. With PPy-N as the electrodes, the assembled supercapacitor possessed a high cyclic stability (88.9% capacitance retention after 1000 cycles) as well as an outstanding flexibility (only 9.6% capacitance decay when bent to 180°) at −50 ℃. Furthermore, the PPy-N supercapacitors in series stably powered electronic devices even when repeatedly bent in a dry ice bath. Such remarkable performances make this flexible PPy-N electrode a promising candidate for the LFSCs.

Assessment of hemodynamics in patients with hypertension using dry carbon dioxide baths and ozone therapy

BACKGROUND: Despite the wide prevalence of arterial hypertension in modern society, including in the Russian population, patients sometimes do not suspect the presence of the disease, and among patients with an established diagnosis, drug therapy has low effectiveness.&#x0D; AIMS: Of the study was to study the effect of dry carbon dioxide baths and ozone therapy on heart rate variability and reocardiography in patients with hypertension.&#x0D; MATERIALS AND METHODS: We observed 71 patients with stage I hypertension and grade 12 arterial hypertension. Patients were randomly selected into three groups: in the control group (n=18), patients received complex spa treatment, including diet, physical therapy, massage of the collar area and aromatherapy; in the comparison group (n=20) basic spa therapy and dry carbon dioxide baths; in the main group (n=33) basic spa therapy, ozone therapy and dry carbon dioxide baths.&#x0D; RESULTS: The combined use of dry carbon dioxide baths and ozone therapy in the complex treatment of patients with hypertension in sanatorium conditions is accompanied by a significant improvement in heart rate variability and central hemodynamics.&#x0D; CONCLUSION: Complex spa treatment of patients with hypertension with the additional appointment of dry carbon dioxide baths and ozone therapy helps to reduce the tension of the central mechanisms of regulation of the cardiovascular system, restore vegetative balance and vascular tone.

The Effect of Dry Carbon Dioxide Bathing on Peripheral Blood Circulation Measured by Thermal Imaging among Patients with Risk Factors of PAD.

Peripheral artery disease (PAD) is becoming a serious health problem of present times. It appears crucial to explore therapies that might help to restore blood flow or increase tissue oxygenation. The most effective methods of detecting early-stage changes in blood circulation in the extremities need to be identified. The aim of this study was to identify the effect of carbon dioxide (CO2) bathing on peripheral blood circulation measured by thermal imaging among patients with risk factors of PAD and ankle–brachial index (ABI) in the normal range or ABI indicating some or moderate arterial disease (ABI > 0.5). The correlation between surface temperature change and PAD-relevant characteristics was also examined. Forty-six patients who were over 65 years old who had a minimum of two additional PAD risk factors were recruited. A series of ten dry CO2 baths was performed. Thermal images were taken before and after the intervention. The CO2 therapy caused a significant change in the body surface temperature of many body areas. Numerous moderate correlations between temperature change and health-related characteristics were identified. Therefore, patients with PAD risk factors could benefit from CO2 therapy. Improvements in blood flow change the body surface temperature, and these changes could be successfully detected by thermal imaging.

Changes in body temperature of critically ill patients submitted to bed bathing: a crossover clinical trial.

to compare tympanic and axillary body temperature values of critical patients before and after the traditional and dry bed bath. this is a randomized, open crossover clinical trial conducted with 50 adult critical patients. All patients received both types of bed bathing. The tympanic and axillary temperature values were measured at the beginning and end of the baths. The Wilcoxon test or paired Student's t test was used. elderly and male patients predominated. There was no significant difference between tympanic temperature medians measured during the traditional bed bath (p=0.707) and dry bath (p=0.101). Axillary temperature means reduced at the end of the baths (p=0.001), being 36.12ºC in the traditional bath and 35.92ºC in dry bath. bed bath, regardless of the method used, caused a reduction in critical patients' axillary temperature.

Comparative analysis of the influence of dry sauna bathing and whole-body cryostimulation on motor nerve excitability

Comparative analysis of the influence of dry sauna bathing and whole-body cryostimulation on motor nerve excitability

Sweating as a Preventive Care and Treatment Strategy in Traditional Persian Medicine.

Proper and regular sweating plays a significant thermoregulatory role. It is a common perception that, sweating has other important homeostatic functions such as clearance of excessive micronutrients, waste products of metabolic processes, and toxins from the body, which helps to maintain human good health. In addition, sweating, thermotherapy, and sauna are commonly used to treat various diseases such as cardiovascular, respiratory and joint diseases. In traditional Persian medicine (PM) textbooks, sweating is considered a preventive care and treatment strategy as well. In this study, we aim to explain the beneficial effects of sweating in human health and its role in the management of various diseases, as well as introducing the therapeutic applications of some diaphoretic plants from the viewpoint of PM. We reviewed the most famous PM textbooks such as Kamil al-Sinaa al-Tibbiya, Al-Qānūn fī al-Tibb, Zakhireye Kharazmshahi, Kholasat al-Hikmat, Exir-e-Azam, and Hifzos-sihhat-e Naseri. Also, current evidence was searched in PubMed, Web of Science, Scopus, and other relevant databases related to the topic. The results of this study revealed that PM scientists believed proper sweating removes waste products and maintains the body’s health, thus, any disturbances in the excretion of these waste products can cause diseases. They recommended the induction of sweating through hot and dry baths, sun bath, sand bath and also the use of diaphoretic herbs for the management of various diseases. Therefore, further researches are recommended to evaluate the effectiveness of these diaphoretic plants.

Modeling Burn Progression Using Comb Burns: The Impact of Thermal Contact Duration on Model Outcomes

BackgroundBurn progression is a phenomenon that remains poorly characterized. The mechanisms of burn conversion are not completely understood, and consequently, both predictive diagnostic tools and interventions are limited. The rat comb burn model is a commonly used approach to study horizontal burn conversion. However, there is significant variability in how the model is performed. Skin contact duration, comb device heating method, comb heating duration, amount of pressure applied, the weight of the comb, and associated depth of burn are all variables that are heterogeneous in studies utilizing the model. Materials and methodsHere, contact duration was examined to determine the impact the duration of burn delivery has on the conversion of interspaces in this model. Data from multiple experiments consisting of 10, 15, 20, 30, 40, and 45 s comb burns were compiled and assessed. Burns were made using combs heated in a 100°C dry bath and then monitored for 2 d. Interspace viability was assessed by digital and laser doppler imaging and biopsy procurement. ResultsLaser Doppler Imaging and viable interspace measurements showed that as burn duration increased, the percentage of the viable interspace and interspace perfusion decreased. Additionally, a contact time of 30 s or greater was required to result in 100% interspace conversion. ConclusionsThese results demonstrate a need to better characterize and potentially standardize the rat comb burn model to reduce variation and maintain it as a valuable tool for controlled studies of the pathophysiology of burn wound progression.

Comparison of Methods of Pollen Selection for Heat Tolerance and Their Effect in Segregating Population of Maize (Zea mays)

The reproductive stage in many crops including Maize (Zea mays L.) is most sensitive to heat stress. Due to limitations in sporophytic selection, an alternative pollen selection approach opens a way for breeders to develop heat stress resilient lines. In present study, the efficiency of different methods of pollen selection for heat stress tolerance was evaluated. The effect of selection in F1 generation in terms of the frequency of heat-tolerant plants in F2 generation was assessed. The heat susceptible line BTM4 was crossed to heat-tolerant BTM6, and the true F1 plants were identified using two SSR markers. Three different methods of pollen selection for heat tolerance, viz. incubating the freshly dehisced pollen grains of F1 plants at 36 °C for 3 h in growth chamber (GRC), dry bath (DB) before selfing and growing F1 plants in summer at ARS, Bheemarayanagudi (BGD). Thus, selected F2 from different treatments was produced. The control F2 was also produced without heat treatment of pollen grains. The control and selected F2 population were compared for heat tolerance by selfing the F2 plants with heat-stressed pollen grains. The selected F2 (GRC) showed superiority over other methods for seed yield traits and its component traits and also positive effect of pollen selection for heat tolerance.

The Use of Dry Carbon Dioxide Baths in the Medical Rehabilitation of Children with Various Diseases

The purpose of this article is systematization and analysis of the results of studies on the effectiveness of using dry carbon dioxide baths in the medical rehabilitation of children with various diseases. Dry carbon dioxide baths allow to dose carbon dioxide during the procedure. The method eliminates the hydrostatic component and excludes the penetration of carbon dioxide into the respiratory tract during the procedure. The article presents the main aspects of the formation of the mechanism of the therapeutic action of dry carbon dioxide baths in bronchial asthma, atopic dermatitis, vegetative dystonia syndrome, health improvement of children who often suffer from acute respiratory diseases. The use of dry carbon dioxide bath in medical rehabilitation of children with bronchial asthma helps to reduce the intensity of allergic inflammation, bronchial obstruction, the frequency of exacerbations of the disease. The use of SUV in atopic dermatitis in children allows you to stop or reduce the manifestations of the main clinical symptoms of the disease, improves the psycho-emotional state of the child. The effectiveness of dry carbonic bath in the vegetative dystonia is shown (mainly in the sympathicotonic type). The improvement of autonomic regulation and the reduction of tension in the work of the sympathetic department of the autonomic nervous system have been established. The application of dry carbon dioxide bath in sickly children helps to reduce clinical symptoms of acute respiratory infections, improves mucosal immunity, and reduces the frequency of recurrent infections throughout the year.

820 Filling Injection of Platelet-rich Plasma Gel as A New Method to Treat En Coup de Sabre Scleroderma

Platelet-rich Plasma (PRP) plays an important role on the tissue regeneration process. A new preparatioin method of PRP provides an filling injection product which was named Platelet-rich Plasma Gel (PRPG) and could provied synergic effects on skin elasticity, dermal remodeling and re-vascularization. The aim of this pilot study was to assess the clinical benefit of PRPG filling injection on the treatment of en coup de sabre scleroderma. Five patients who were diagnosed under the confirmation of histopathology, and assessed by ultrasonography. Treatment was administered with PRP at a concentration 3.5 times above baseline. And under sterile condition, pre-seperated PRP was aspirated into several needle tubes, gradually heated in a dry bath machine, and prepare PRPG under the terminal temperature of 70 centigrade for 14 minutes. Under swelling anesthesia, subcutaneous tissue gaps were formed. Then PRPG was safely filled in these gaps without the risk of injection into the blood vessels. Therefore, a kind of structure mimic subcutaneous soft tissue was performed to slowly release cell and growth factors which enhance the regeneration effects on the remodeling of dermal tissue and reconstruction of vascular network. The subjective symptoms of pain and tightness were significantly relieved after at least one time of injecton. And after completion of 3 cycles of treatments at monthly intervals, objective lesions showed the improvement of skin elatisity, dermal thickness, subcutaneous tissue thickness and subcutaneous vascular network density. Even hair growth can be observed in those atrophy scalp filed. The main limitations of this study were the low number of cases, lack of a comparison group and the difficulty of identifying the exact role of each factor which would be studied in next clinical trial. However, PRPG is a good alternative of tissue filler substitution instead of hyaluronic acid and collagen, and represents a new and potentially regenerative treatment option for en coup de sabre scleroderma with minimal associated side effects.

Dry carbon dioxide baths in complex spa treatment of children and adolescents with diseases of the cardiovascular system

These guidelines provide a scientifi c justifi cation for the use of dry carbon dioxide baths in the General complex of Spa treatment of children and adolescents with diseases of the cardiovascular system. The guidelines are intended for doctors of medical rehabilitation, pediatricians, therapists, cardiologists, physiotherapists of specialized medical and preventive institutions, rehabilitation departments and centers, sanatoriums, and dispensaries.

Does repeated dry sauna bathing change thermoregulation process in elite cross-country skiers?

The aim of this study was to evaluate the influence of a series of ten finnish dry sauna bathing on skin temperature following exercise test for elite cross-country skiers and test, if the regular sauna baths induce any changes in physiological or haematological parameters in elite athletes. Ten elite cross-country skiers (21.7 ± 3.1 years, height: 1.79 ± 0.025 m, body mass: 71.06 ± 4.77 kg) participated in this study. They performed two continuous exercise tests, before and after a series of ten sauna baths. During experiment, the superficial temperature distributions, physiological and haematological parameters were measured. The marked temperature drop was visible in upper body and smaller in lower part of body after exercise test. 10 min after experimental exercise test slow return of the skin temperature to pre-workout values was observed in lower limbs unlike upper part of body. Results of this study indicated that passive induced heat acclimation have a moderate, positive effect (above 2%) on plasma volume and a small impact on physiological and temperature responses to exercise in elite athletes. We conclude that ten finnish dry sauna baths can induce weak changes in the superficial temperature distributions following exercise test, a certain decrease in resting heart rate and small increase in plasma volume in the elite cross-country skiers.

Thawing cryopreserved apheresis products: comparison of barkey plasmatherm dry heating device and 37oC water bath

Background & Aim The Barkey Plasmatherm (Barkey GmbH & Co. KG) dry heating device (BPDHD) is designed to thaw frozen products intended for infusion. This study compared cryopreserved apheresis hematopoietic stem/progenitor cell (HS/PC) product CD34 + cell dose recovery, viability and sterility using the BPDHD and a traditional 37°C water bath (Isotemp; Fisher Scientific). Methods, Results & Conclusion METHODS Cryopreserved HS/PC apheresis products were used to assess CD34 recovery, viability and sterility. We selected two products from a single processing and freezing from three independent collections. A paired bag from each of the three collections and freezing procedures was thawed using either the BPDRD or Isotemp water bath. Each thawed product was tested for CD34+ cell dose recovery, CD34+ cell 7AAD viability, sterility by bacterial culture, and colony forming unit (CFU) growth (Stem Cell Technologies). We also performed an independent 3-month post-transplant follow up to compare the time to neutrophil and platelet engraftment in patients who received products thawed using the BPDHD vs products thawed using the Isotemp water bath RESULTS Three products were thawed using BPDHD and 3 products were thawed using a water bath for a total of 6 products thawed and tested. Table 1 summarizes the post-thaw testing performed on each product. All post-thaw testing met our established release criteria (Table 2). We compared the engraftment outcomes for the first 15 patients who received products thawed with the BPDHD to transplant recipients who received products thawed in the Isotemp water bath. We evaluated days to > 500 neutrophils/uL for three consecutive days, and days to platelet count > 20 × 109/L without platelet transfusion support for 7 days. We found no difference between the two thawing methods (Table 3.) CONCLUSION A 37oC water bath is the most commonly used device to thaw cellular therapy products in the United States. This method requires manually submerging the product directly into temperature-controlled water and massaging the bag to ensure a complete thaw, and the water must be changed after each use. The BPDHD is a dry heating system with water-filled cushions and the water is changed annually. Moreover, the water never touches the product, which in principle will reduce the possibility of contamination. The BPDHD takes away the manual manipulation of the product making thawing more standardized, which may translate into improved laboratory efficiency. The Barkey Plasmatherm (Barkey GmbH & Co. KG) dry heating device (BPDHD) is designed to thaw frozen products intended for infusion. This study compared cryopreserved apheresis hematopoietic stem/progenitor cell (HS/PC) product CD34 + cell dose recovery, viability and sterility using the BPDHD and a traditional 37°C water bath (Isotemp; Fisher Scientific). Cryopreserved HS/PC apheresis products were used to assess CD34 recovery, viability and sterility. We selected two products from a single processing and freezing from three independent collections. A paired bag from each of the three collections and freezing procedures was thawed using either the BPDRD or Isotemp water bath. Each thawed product was tested for CD34+ cell dose recovery, CD34+ cell 7AAD viability, sterility by bacterial culture, and colony forming unit (CFU) growth (Stem Cell Technologies). We also performed an independent 3-month post-transplant follow up to compare the time to neutrophil and platelet engraftment in patients who received products thawed using the BPDHD vs products thawed using the Isotemp water bath

Effect on Skin Microflora in Hygienic Bathing of Patients with Chlorhexidine Gluconate Disinfectant and Non-Rinse Product

Objective: The skin is one of the largest reservoirs of microorganisms (MO) in the human body, some of which are potential agents of Health care-associated infections (HAIs). The purpose of the present study is to provide a comparative microbiological analysis of the skin surface in patients before and after the application of the dry-bath method with an antiseptic agent-chlorhexidine gluconate (CHG) and with non-rinsing detergents. The method is innovative for the Republic of Bulgaria and has not been imposed and has not yet been implemented as a routine practice in hospitals. The purpose of this study is to demonstrate the bacterial contamination of the skin after a second day of hospital stay and the need to use agents containing sanitary disinfectant for patients. Methods: Methods based on bacterial culture using swabs were used. These are one of the most commonly used methods for isolating and identifying aerobic and optional anaerobic MO that are part of the skin flora. Results: The following bacteria were isolated in the microbiological examination of the skin surface: Coagulasenegative methicillin sensitive staphylococci (CNMSS), Coagulasenegative methicillin resistant staphylococci (CNMRS), Enterococcus, Methicillin-sensitive S. aureus (MSSA), Bacillus spp, Gram/-/bacteria, Streptococcus α-hem, Corynebacterium. Conclusion: The present studies demonstrate a significant decrease in microbial flora after CHG administration and a slight to no effect after applying non-rinsing detergents. The research team's recommendations are to apply products with CHG from nurse for hygiene of patients by the method of dry bathing at risk of HAIs.

Influential Factors in the Preparation of 68Ga-DOTATATE.

Acceptable and reproducible radiochemical purity (RP) for 68Ga-DOTATATE was difficult to obtain with the NETSPOT kit because the manufacturer instructions lacked details on the heater or needles used. Methods: The drug was prepared in an International Organization for Standardization (ISO) 5 environment. Multiple dry baths and needle types were used to investigate the effects of reaction temperature and metal contamination, respectively. Temperature curves were obtained with a calibrated thermocouple. The influence of the accuracy of the NETSPOT reagent volume and its effect on outcome were investigated. Results: The AccuBlock dry bath required recalibration for the ISO 5 environment; after calibration, the temperature was stable (only ±0.1°C from the set point). When we followed package insert recommendations (dry bath temperature set to 98°C, reaction time of 8 min), the reaction temperature was 90.6°C. When Becton Dickinson needles were used for reconstitution, 15 of 18 runs (83%) did not meet the RP specification. However, B. Braun Medical needles achieved satisfactory and stable RP. When the 68Ga generator was eluted with 5.0 mL of 0.1 M hydrochloric acid (HCl), only 3.8-3.9 mL of eluate reached the reaction vial; this volume did not impact labeling (final pH was 3.8). The labeling success rate increased markedly if the 68Ga eluate was passed through a conditioned silica gel cartridge or if no cartridge was used; then, RP was more than 99%. HCl contact with the septum of the labeling vial reduced RP. Conclusion: The needle type and the temperature setting of the dry bath have critical roles in 68Ga-DOTATATE preparation. The AccuBlock dry bath has excellent stability and accuracy and can be used for reliable preparation. By using a conditioned silica gel cartridge or by eliminating the cartridge altogether, the RP is reliably high and stable.

Effect of virus inactivation on weak positive results of nucleic acid test for 2019 novel coronavirus

Objective To investigate the effect of virus inactivation on weak positive result of 2019 Novel Coronavirus (2019-nCoV) nucleic acid test. Method A retrospective study was conducted on the nasopharyngeal swabs of three patients with positive PCR nucleic acid test for Novel Coronavirus at different concentrations in the Second affiliated Hospital of Zhejiang University Medical College from January to February 2020.The virus in nasopharyngeal swab specimens were inactivated by water bath at 56 ℃ for 30 min, dry bath at 56 ℃ for 60 min and dry bath at 60 ℃ for 30 min respectively. After treatment, the sample RNA were extracted and then detected by three new commercial quantitative real-time polymerase chain reaction reagent kits for 2019-nCoV.Cycle threshold(Ct)value was used to evaluate the effect of virus inactivation on nucleic acid detection of 2019-nCoV. Results There was no significant difference between the groups before and after inactivation. Ct values of ORF1ab gene before inactivation were 23.28±0.28, 25.25±0.25, 28.93±0.44, 32.06±0.47, 35.20±0.38, 32.89±0.38, 36.24±0.23, 33.30±0.46, and those after inactivation were, group 1:23.60±0.20, 27.29±0.30, 31.83±0.51, 37.41±0.46, group 2:24.25±0.34, 27.18±0.42, 31.84±0.61, 34.99±1.01, 34.89±0.45,group 3:23.37±0.17, 26.89±0.52, 32.05±0.50.Ct value of N gene before inactivation were24.38±0.09, 26.64±0.11, 30.35±0.12, 33.29±0.33, 36.93±0.11, 34.50±0.12, 35.63±0.12, those after inactivation were, group 1:24.66 ±0.11, 28.52±0.14, 32.71±0.14, 37.00±0.13.group 2:25.41 ±0.10, 28.79±0.15, 33.29±0.28. group 3:23.37±0.11, 28.68±0.11, 33.54±0.13, 37.18±0.23(ORF1ab gene: t =-1.416; N gene: t=-1.379, P> 0.05). There was no significant difference amongthe three inactivation groups, the specific Ct values are shown above(ORF1ab gene: t =-0.460; N gene: t =-0.132, P>0.05). However, the Ct values of the inactivated groups (1,2,3) and the non-inactivated group at different dilution times were different (10 ×:Ct value of ORF1abwas 25.25±0.25 in the non-inactivated group, and 27.29±0.30, 27.18±0.42 and 26.89±0.52 in the inactivated group1,2 and 3.t (ORF1ab) = -7.327, P 0.05) and among the three reagents(reagent 1:7/11, 4/11, 3/11, 3/11, reagent 2:8/11, 4/11, 3/11, 3/11, reagent 3:5/11, 3/11, 3/11, 2/11)(χ2=1.199, P>0.05). Conclusion The virus inactivation can degrade the nucleic acid of the 2019nCoV, resulting in the decrease of the Ct value and the false negative results of the low-concentration specimens. Key words: Coronavirus; Virus inactivation; Nucleic acids; False negative reactions; Real-time polymerase chain reaction

Passport and biological age in the choice of metabolic geroprophylactic therapy

We compared the effectiveness of geroprophylactic means of gas and metabolic therapy (hypoxytherapy, hyperbaric oxygenation, ozone therapy, dry carbonic baths, amino acid arginine, regulatory tripeptides H- Glu-Asp-Arg-OH u H-Lys-Glu-Asp-OH, interleukin-2 in medium therapeutic dosages by their ability to reduce biological age when tested on practically healthy people or patients with mild subclinical forms of polymorbid pathology at the age of 35 – 74 years. It has been established that the effectiveness of agents and effects in terms of the degree of decrease in biological age depends on the type of geroprophylactic agent, as well as in the part of research – the passport age of patients. In this case, the biological age of the subjects after treatment decreased in the range from 12.8 to 1.3 years, depending on the means used and, in some cases, on the passport age. The greatest efficiency was achieved after the use of a combination of peptide bioregulators in the age group with a passport age of 35 – 59 years. When prescribing metabolic geroprophylaxis, it is necessary to take into account, in addition to the indications and contraindications of the means used or the correction method, the state of the body, also the calendar age and the degree of decrease in biological age.

Immunohistochemistry (IHC): Chromogenic Detection of 3-Phosphohistidine Proteins in Formaldehyde-Fixed, Frozen Mouse Liver Tissue Sections.

The development of antibodies that specifically detect histidine-phosphorylated proteins is a recent achievement and allows potential roles of histidine phosphorylated proteins in pathological and physiological conditions to be characterized. Immunohistochemical analyses enable the detection of proteins in tissues and can reveal alterations to the quantity and/or localization of these proteins through comparisons of normal and diseased specimens. However, the sensitivity of phosphohistidine modifications to phosphatases, acidic pH, and elevated temperatures poses unique challenges to the detection process and requires a protocol that bypasses traditional procedures utilizing paraffin-embedding and antigen-retrieval methods. Here, we detail a method for a brief fixation by 4% (v/v) paraformaldehyde on freshly collected tissues in the presence of PhosSTOP to block phosphatase activity, followed by a float on sucrose to protect the tissue prior to freezing. Specimens are then embedded in a cryopreservation medium in molds and frozen using an isoflurane, dry ice bath to best preserve the tissue morphology and phosphohistidine signal. We validate this technique in normal mouse liver using SC44-1, a monoclonal anti-3-pHis antibody used to uncover a role for a protein histidine phosphatase as a tumor suppressor in the liver. Furthermore, we demonstrate that the antibody signal can be eliminated by preincubating SC44-1 with a peptide treated with phosphoramidate to phosphorylate histidine residues. Thus, we present an IHC protocol suitable for specific detection of 3-phosphohistidine proteins in mouse liver tissue, and suggest that this can be used as a starting point for optimization of IHC using other phosphohistidine antibodies or in other tissue types, generating information that will enhance our understanding of phosphohistidine in models of disease.

Rapid authentication of mutton products by recombinase polymerase amplification coupled with lateral flow dipsticks

Shish kebabs and mutton slices are two of the most favorite foods in China, which are often adulterated with low-cost animal meat such as chicken or duck meat. Currently, many methods have been applied to mutton adulteration detection. However, existing methods based on protein or polymerase chain reaction (PCR) are time-consuming and tedious, making them unfit for rapid and on-site detection of counterfeit mutton products in restaurants. In this study, a visual lateral flow dipstick (LFD) method in combination with recombinase polymerase amplification (RPA) was developed for authentication of mutton slices or heat-processed shish kebabs. In this method, sheep-specific RPA primer was designed against the cytochrome b (Cyt b) gene. RPA reaction could be performed under 37–42 °C and finished within 15 min. Then, the products were added onto the LFD sample pad until the final results could be observed by naked eyes within 5 min. The total time for authenticating mutton products was only about 30 min, including 5 min for DNA extraction. In addition, only a portable and inexpensive dry bath incubator was required for the authentication. The proposed method was therefore a simple and fast approach, which was also specific and sensitive, and it could be used for on-site mutton product fraud detection by food safety inspection authorities.

Rapid and Sensitive Detection of Citrus tristeza virus Using Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) Assay.

Loop-mediated isothermal amplification (LAMP) is one recently developed gene amplification technique that emerges as a simple and quick diagnostic tool for early detection of nucleic acid targets. The LAMP technique works on the principle of strand displacement activity of Bst polymerase. It contains a set of four specially designed primers, which recognizes six different regions on the target nucleotide sequence. In the LAMP reaction, amplification is carried out in an isothermal conditions (60-65°C) using simple and inexpensive device like water bath or dry bath. Additional benefits of LAMP technique are that final results can be seen directly with naked eyes by adding intercalating dye SYBR Green I in the reaction tube. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) is one of the novel techniques used for detection of RNA targets. The technology has been successfully applied for rapid and sensitive detection of Citrus tristeza virus (CTV) by using four oligo-primers, targeting a conserved coat protein gene (CPG) of an Indian CTV isolate. The result of assay is visible in naked eyes easily in the presence of SYBR Green I (100×) or on 1.5% agarose gel electrophoresis. CTV-RT-LAMP could be used away from plant pathology laboratories even in remote location.