mL Of Volume Research Articles

The Efficacy and Safety of Dr. SKS Hair Booster Serum (a Cocktail of Micronutrients and Multivitamins) in Adult Males and Females With Androgenetic Alopecia: An Open-Label, Non-randomized, Prospective Study.

Background Androgenetic alopecia (AGA) is frequently encountered in dermatological practice; however, there is a lack of approved treatment. At present, only three therapies have been approved for on-label use in androgenetic alopecia: minoxidil, finasteride, and lower-level laser therapy. Micronutrients are primary elements in the normal hair follicle cycle, and their role in androgenetic alopecia is a growing matter of research nowadays. This study aims to investigate the clinical efficacy and safety of Dr. SKS Hair Booster Serum, a cocktail of micronutrients and multivitamins (copper, niacinamide, hyaluronic acid, thiamine, riboflavin, and biotin), in male and female patients with androgenetic alopecia. Methods We did an open-label, non-randomized, multicenter, prospective study across five hair clinic chains in India (Mumbai, Hyderabad, Jabalpur, Balaghat, and Nagpur). Eligible participants were patients with a confirmed diagnosis of androgenetic alopecia based on clinical examination and trichoscopic findings, age of 18 years or older, and any gender. Each patient received Dr. SKS Hair Booster Serum, 1 ml in volume, once a month by mesotherapy or derma roller/derma pen for up to six months. All patients were subjected to a 60-second hair counttest (comb test), hair pulltest, global photographic assessment (GPA), trichoscopy assessment, patient self-assessment questionnaire, and safety assessment at baseline and six months after the treatment. Results One thousand patients (500 males and females each) with androgenetic alopecia were analyzed. There was a significant reduction in hair fall with bulb (<0.0001) and without bulb (<0.0001) six months after the treatment versus baseline. There was a significant improvement in the number of hairs removed per pull (<0.0001), global photographic assessment score (<0.0001), hair growth rate (<0.0001), follicular hair density (<0.0001), vellus hair density (<0.0001), and terminal hair density (<0.0001) six months after the treatment versus baseline. The majority of patients (95%) were satisfied with six-monthtreatment of Dr. SKS Hair Booster Serum. No major adverse events were reported during the study. Conclusion Dr. SKS Hair Booster Serum was found to be a safe and effective treatment for androgenetic alopecia, with 95% patient self-assessment score.

Effects of the In ovo Administration of L-Ascorbic Acid on Tissue L-Ascorbic Acid Concentrations, Systemic Inflammation, and Tracheal Histomorphology of Ross 708 Broilers Subjected to Elevated Levels of Atmospheric Ammonia

The effects of in ovo injection of L-ascorbic acid (L-AA) on tissue L-AA concentrations, systemic inflammation, plasma mineral concentrations, and tracheal histomorphology of Ross 708 broilers subjected to elevated atmospheric ammonia (NH3) levels after hatch were investigated. The four in ovo treatments included non-injected (control), saline-injected (control), or saline containing 12 or 25 mg of L-AA. The in ovo treatments were applied at 17 days of incubation by injecting a 100 μL volume of each pre-specified treatment into the amnion. At hatch, 12 male chicks were randomly allocated to each of the 12 replicate battery cages belonging to each treatment group. The cages were arranged in a randomized complete block design within a common room. All birds were exposed to 50 ppm of NH3 at 35 days of posthatch age (doa), and the concentration of NH3 in the room was recorded every 20 s. At 0, 7, 14, 21, and 28 doa, one bird from each cage was arbitrarily selected and euthanized for determinations of liver and eye L-AA concentrations at 0, 7, 14, 21, 28 doa; plasma nitric oxide concentrations at 0, 14, 21, and 28 doa; and plasma calcium and trace mineral concentrations at 0 and 21 doa. Tracheal histomorphology evaluations were performed at 0, 21, and 28 doa. There were no significant treatment differences for plasma nitric oxide and mineral concentrations, and for liver and eye L-AA concentrations at each sampling timepoint. In ovo injection of either 12 or 25 mg of L-AA decreased tracheal attenuation incidence at 0 doa compared to the non-injected or saline-injected control groups. Furthermore, the percentage of mild tracheal inflammation scores was lower at 28 doa in response to the in ovo injection of 12 mg of L-AA compared to the non-injected or saline-injected control groups. These results indicate that in ovo injection of 12 mg of L-AA reduces tracheal inflammation in broilers subjected to elevated atmospheric NH3.

In Vitro Antibody Quantification with Hyperspectral Imaging in a Large Field of View for Clinical Applications.

Hyperspectral imaging (HSI) is a non-invasive, contrast-free optical-based tool that has recently been applied in medical and basic research fields. The opportunity to use HSI to identify exogenous tumor markers in a large field of view (LFOV) could increase precision in oncological diagnosis and surgical treatment. In this study, the anti-high mobility group B1 (HMGB1) labeled with Alexa fluorophore (647 nm) was used as the target molecule. This is the proof-of-concept of HSI's ability to quantify antibodies via an in vitro setting. A first test was performed to understand whether the relative absorbance provided by the HSI camera was dependent on volume at a 1:1 concentration. A serial dilution of 1:1, 10, 100, 1000, and 10,000 with phosphatase-buffered saline (PBS) was then used to test the sensitivity of the camera at the minimum and maximum volumes. For the analysis, images at 640 nm were extracted from the hypercubes according to peak signals matching the specificities of the antibody manufacturer. The results showed a positive correlation between relative absorbance and volume (r = 0.9709, p = 0.0013). The correlation between concentration and relative absorbance at min (1 µL) and max (20 µL) volume showed r = 0.9925, p < 0.0001, and r = 0.9992, p < 0.0001, respectively. These results demonstrate the HSI potential in quantifying HMGB1, hence deserving further studies in ex vivo and in vivo settings.

New Method for Counting and Picking Out Single Circulating Tumor Cells from Microliter-Volume Samples for Tumor Progression Surveillance and Single-Cell Heterogeneity Analysis

Circulating tumor cells (CTCs) are crucial in tumor progression and metastasis, but the knowledge of their roles grows slowly at single-cell levels. Characterizing the rarity and fragility of CTCs by nature, highly stable and efficient single-CTC sampling methods are still lacking, which impedes the development of single-CTC analysis. Herein, an improved, capillary-based single-cell sampling (SiCS) method, the so-called bubble-glue single-cell sampling (bubble-glue SiCS), is introduced. Benefiting from the characteristic that the cells tend to adhere to air bubbles in the solution, single cells can be sampled with bubbles as low as 20 pL with a self-designed microbubble-volume-controlled system. Benefiting from the excellent maneuverability, single CTCs are sampled directly from 10 μL volume of real blood samples after fluorescent labeling. Meanwhile, over 90% of the CTCs obtained survived and well proliferated after the bubble-glue SiCS process, which showed considerable superiority for downstream single-CTC profiling. Furthermore, a highly metastatic breast cancer model of the 4T1 cell line in vivo was employed for the real blood sample analysis. Increases in CTC numbers were observed during the tumor progression process, and significant heterogeneities among individual CTCs were discovered. In all, we propose a novel avenue for target SiCS and provide an alternative technique route for CTC separation and analysis.

Fluorescent single particle tracking enables high sensitivity viscometry measurements of micro-volume samples.

Measuring the viscosities of high-value materials at the microscale with minimal waste is essential for increasing turnaround times during drug development in the pharmaceutical industry. Currently, viscometers on the market fail to meet these needs as they can only process a limited number of samples at a time and still require large (>100 μL) loading volumes for accurate readings. Previously, researchers have parameterized fluorescence correlation spectroscopy (FCS) at low volumes to predict antibody behaviors at higher concentrations using the generalized Stokes-Einstein equation. However, these methods lack the ability to produce measurements at a fast turnaround rate. Here, we use a wide field camera-based fluorescence microscope modality and particle tracking system in parallel with a Python PT software to measure the viscosities of multiple solutions using a 10-μL working volume. We applied the generalized Stokes-Einstein equation to the coefficient output of the PT Python software to obtain our final viscosity values. We obtained values of viscosity with an overall error of +-4cP, and the sample preparation method resulted in consistent readings for up to 5 hours after initial preparation. Viscosity readouts obtained using this method will allow for multi-sample imaging on a single chip using minuscule (10 μL) volumes, revolutionizing the efficiency of sample selection during drug development.

Constriction of H2 biofuel cell voltammeter based on Escherichia coli microbial cells.

Hydrogen (H2) is the lightest and most abundant chemical element in the universe, with potential to replace fossil fuels as a carbon-free source of energy. Sensitive and faster H2 gas sensors are/will be required in many industries and hydrogen-based infrastructures. Microbial hydrogenases (Hyds) reversibly catalyze the formation and oxidation of H2. Isolation and characterization of O2-tolerant [NiFe]-Hyds have stimulated new concepts in hydrogen biofuel cells (BFCs). Platinum-independent H2-driven bio-electrochemical devices are suitable for environmentally friendly energy generation and application in biosensors. Escherichia coli employs four [NiFe]-Hyds, which can be applied as anode biocatalysts in BFCs. In this study, the H2 BFC voltammeter working with graphite based strips (sensors) were constructed and the efficiency of applying the E. coli BW25113 whole cells or crude extracts as anode biocatalysts were evaluated. E. coli BW25113 was grown under fermentation of 0.2% glucose or 1% glycerol in a peptone medium, pH 7.5. Upon glucose fermentation, H2-oxidizing Hyd activity of E. coli whole cells was 1.03±0.01 U (mg CDW)−1 and 1.39±0.01 U (mg CDW)−1 was observed in cell extracts. During glycerol fermentation, Hyd activity reached up to 2.15±0.02 U (mg CDW)−1 and 3.58±0.02 U (mg CDW)−1, respectively in whole cells and cell extracts. 3 μL volumes of bacterial whole cells or cell extracts were immobilized on the sensors with the aid of 3 mM KCl and 0.8% bacteriological agar support. Whole immobilized cells(grown on glucose) produced an electrical potential of 0.650±0.05 V, and with the cell extracts, the electrical potential was slightly lower. Meanwhile, under glycerol fermentation, the electrical potential was 0.550±0.05 V in cell extracts and whole cells. Despite, the highest electrical potential was achieved with bacterial whole cells grown on glucose, H2-oxidizing Hyd activity was stimulated upon bacterial glycerol fermentation.

Porous PDMS‐Based Microsystem (ExoSponge) for Rapid Cost‐Effective Tumor Extracellular Vesicle Isolation and Mass Spectrometry‐Based Metabolic Biomarker Screening

AbstractPolydimethylsiloxane (PDMS) is an inexpensive robust polymer that is commonly used as the fundamental fabrication material for soft‐lithography‐based microfluidic devices. Owing to its versatile material properties, there are some attempts to use PDMS as a porous 3D structure for sensing. However, reliable and easy fabrication has been challenging along with the inherent hydrophobic nature of PDMS hindering its use in biomedical sensing applications. Herein, a cleanroom‐free inexpensive method to create 3D porous PDMS structures, “ExoSponge” and the effective surface modification to functionalize its 3D porous structure is reported. The ability of ExoSponge to recover cancer‐associated extracellular vesicles (EVs) from complex biological samples of up to 10 mL in volume is demonstrated. When compared to ultracentrifugation, the ExoSponge showes a significant increase in cancer EV isolation of more than 210%. Targeted ultra‐high pressure liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) is further employed to profile 70 metabolites in the EVs recovered from the lung cancer patient's plasma. The resulting profiles reveal the potential intraexosomal metabolite biomarker, phenylacetylglutamine (PAG), in non‐small cell lung cancer. The high sensitivity, simple usage, and cost‐effectiveness of the ExoSponge platform creates huge potential for rapid, economical and yet specific isolation of exosomes enabling future diagnostic applications of EVs in cancers.

Ultraminiaturized Microfluidic Electrochemical Surface‐Enhanced Raman Scattering Chip for Analysis of Neurotransmitters Fabricated by Ship‐in‐a‐Bottle Integration

Electrochemical surface‐enhanced Raman scattering (EC‐SERS) is a promising technique for the diagnosis of trace amounts of neurotransmitters, because it can elucidate neurotransmitters’ behavior on electrodes to deduce their functions in the human body. However, the current EC‐SERS devices need several tens of milliliters of analyte solution to collect enough signal for analysis. Miniaturization of EC‐SERS devices is crucial for the early diagnosis of disease and point‐of‐care testing. Herein, a new type of EC‐SERS sensor based on 3D microfluidic chips for the analysis of neurotransmitters in ultrasmall volumes is proposed. The microfluidic EC‐SERS chip is fabricated by a ship‐in‐a‐bottle technique based on hybrid laser processing. The working electrode is modified using silver/zinc oxide materials, enabling the formation of a unique “candy apple” structure. To assess the fabricated microfluidic EC‐SERS chips, ascorbic acid is analyzed using the ingenious microfluidic EC‐SERS chips to elucidate its redox reaction by EC‐SERS spectroscopy. Significantly, a sub‐10 μL volume of analyte solution is sufficient for EC‐SERS analysis, which is several orders smaller in volume than the requirements of current EC‐SERS devices. The unprecedented microfluidic EC‐SERS chips fabricated by the ship‐in‐a‐bottle integration technique can be used in portable and smart analyzers for next‐generation biomedicines and catalysts.

Determination of lithium concentration in capillary blood using volumetric dried blood spots

BackgroundLithium is a cornerstone in the treatment of bipolar disorder and is considered one of the most effective treatments in psychiatry at large. Lithium treatment requires individual dosing with frequent serum concentration measurements due to the narrow therapeutic window and risk of toxicity. There is need for patient-centric methods for lithium monitoring and the use of dried blood spots has recently been proposed for determination of lithium concentration. The purpose of the current study was to assess feasibility of this method by introducing a volumetric technique developed for home-sampling. Materials and methodsLaboratory: Capillary blood was sampled by finger-prick using a volumetric device that collects 10 µL volumes as a dried blood spot. Lithium was measured in the dried blood spots using a validated atomic absorption spectroscopy method. Clinical: Thirty-nine lithium-treated patients were recruited, and dried blood spots and venous blood samples were collected. Routine serum analysis was performed for comparison. ResultsThe range of serum lithium concentrations was 0.41–1.22 mmol/L, and the dried blood spot/serum ratio was 0.78. A strong linear correlation between the two specimens was shown with Pearson’s R = 0.95 (r2 = 0.90). Adding hematocrit as a variable only minimally improved prediction. ConclusionVolumetric dried blood spots is a promising technique for measurement of lithium concentrations. This will enable home-sampling and could potentially save resources, improve compliance, and make treatment safer. This may facilitate the use of lithium treatment in regions where monitoring via venous blood sampling remains difficult. However, the usability of dried blood spots for monitoring lithium treatment longitudinally remains to be examined.

ANALYTICAL METHOD DEVELOPMENT FOR SIMULTANEOUS ESTIMATION OF CIPROFLOXACIN HCL AND OLOPATADINE HCL IN BULK DRUG AND FORMULATION BY RP-HPLC METHOD

A reverse phase chromatography technique to simultaneously estimate ciprofloxacin HCl and olopatadine HCl as bulk drugs and in formulation has been developed. The chromatographic separation of drugs and formulation was accomplished on C18 Agilent Zorbax column (250 x 4.6 mm, 5 µm) using acetonitrile: TFA water (40:60, V/V). A 10 µL volume was injected with flow rate 1 mL min-1, and peaks were detected. The retention periods for ciprofloxacin HCl and olopatadine HCl were 2.92 and 5.10 minutes respectively. The method was linear in the concentration ranges 24-36 µg mL-1 and 56- 84 µg mL-1 for ciprofloxacin HCl and olopatadine HCl, with regression coefficient correlation values of 0.9999 and 0.9996, respectively. Relative standard deviation of precision, accuracy, and ruggedness was below 2 %. Forced degradation studies were performed under different conditions which were in an admissible range. The established method was simple, accurate, and precise. Therefore, it can be applied for the routine quantification and degradation of these drugs and formulation.

A Validated UHPLC–MS/MS Method to Quantify Eight Antibiotics in Quantitative Dried Blood Spots in Support of Pharmacokinetic Studies in Neonates

Conduction of pharmacokinetic (PK) study in pediatric patients is challenging due to blood sampling limits. The dried blood spots (DBS) method represents a potential matrix for microsampling in support of PK studies in children. Herein, we used the Capitainer® qDBS device to develop a DBS method that can collect an exact 10 µL volume of blood on a paper card. This DBS method was developed to simultaneously quantify the concentrations of eight antibiotics, including sulbactam, tazobactam, ampicillin, meropenem, cefotaxime, cefoperazone, piperacillin, and metronidazole using ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). The prepared DBS samples were extracted in methanol containing acetaminophen as the internal standard at 20 °C on a block bath shaker at 500 rpm for 30 min. The extracted antibiotics were eluted on an Acquity UPLC HSS T3 column (2.1 × 50 mm, 1.8 µm) using gradient elution with a total chromatographic run time of 6.5 min. The precursor and product ions of the analytes were detected by use of the multiple reaction monitoring (MRM) mode. No interfering peaks at the respective retention times of the analytes were observed in DBS samples. The lower limits of quantification (LLOQ) for the antibiotics were between 0.25 and 2.0 μg/mL, and satisfactory accuracies (intra/inter-assay bias -16.7 to +13.6%) and precisions (intra/inter-assay coefficient of variations 1.5-15.6%) were obtained for the analytes. As a proof of concept, the method was applied to DBS samples obtained from neonatal patients treated with ampicillin and piperacillin/sulbactam. The DBS method is simple and robust, and it can be used in children with limited blood sampling.

Methodology for the use of Equiverm-2.0% against equine parasite infections

The purpose of the research is to develop a methodology for Equiverm-2.0% paste used against equine parasite infections.The methodology provides a brief description of Equiverm-2.0%, mechanism of its action, pharmaco-toxicological properties, application, slaughter terms, and personal safety measures. Equiverm-2.0% antiparasitic paste consists of the active ingredient, ivermectin, and excipients; it is a light cream-coloured paste with a slight pine odor and sweetish taste. The drug is administered orally at a therapeutic dose of 0.2 mg/kg for the active substance. One gram of paste is equal to 1 mL in volume with 20 mg of ivermectin. Ivermectin in Equiverm-2.0% paste is in a dissolved form, and forms an intermolecular complex. The antiparasitic paste is packaged in disposable polyethylene syringe dispensers of 5 or 10 mL, respectively, 4 to 8 g each, per treated horse weighing 400-800 kg. When administered orally, the paste spreads in the oral cavity and the animal eats it with pleasure due to its sweetish taste. The drug is recommended for registration in the Russian Federation.

Biohydrogen Production in Microbial Electrolysis Cell Operating on Designed Consortium of Denitrifying Bacteria.

This study provides insight into the use of a designed microbial community to produce biohydrogen in simple, single-chamber microbial electrolysis cells (MECs). The ability of MECs to stably produce biohydrogen relies heavily on the setup and microorganisms working inside the system. Despite having the most straightforward configuration and effectively avoiding costly membranes, single-chamber MECs are prone to competing metabolic pathways. We present in this study one possible way of avoiding this problem using characteristically defined, designed microbial consortium. Here, we compare the performance of MECs inoculated with a designed consortium to MECs operating with a naturally occurring soil consortium. We adapted a cost-effective and simple single-chamber MEC design. The MEC was gastight, 100 mL in volume, and equipped with continuous monitoring for electrical output using a digital multimeter. Microorganisms were sourced from Indonesian environmental samples, either as denitrifying bacterial isolates grouped as a designed consortium or natural soil microbiome used in its entirety. The designed consortium consisted of five species from the Pseudomonas and Acinetobacter genera. The headspace gas profile was monitored periodically with a gas chromatograph. At the end of the culture, the composition of the natural soil consortium was characterized by next generation sequencing and the growth of the bacteria on the surface of the anodes by field emission scanning electron microscopy. We found that MEC using a designed consortium presented a better H2 production profile, with the ability of the system to maintain headspace H2 concentration relatively stable for a long time after reaching stationary growth period. In contrast, MECs inoculated with soil microbiome exhibited a strong decline in headspace H2 profile within the same time frame. This work utilizes a designed, denitrifying bacterial consortium isolated from Indonesian environmental samples that can survive in a nitrate-rich environment. Here we propose using a designed consortium as a biological approach to avoid methanogenesis in MECs, as a simple and environmentally friendly alternative to current chemical/physical methods. Our findings offer an alternative solution to avoid the problem of H2 loss in single-chamber MECs along with optimizing biohydrogen production through bioelectrochemical routes.

Comparative pathology and immunohistochemistry of Newcastle disease in domestic chicken (Gallus-gallus domesticus) and Alabio duck (Anas platyrhynchos Borneo).

Newcastle disease is very pathogenic in chickens, whereas in ducks, the disease does not show any clinical symptoms. To compare the clinical symptoms features, pathological lesions, viral distribution, and apoptosis response caused by the Newcastle disease virus (NDV) in domestic chicken and Alabio duck. Forty domestic chickens and 40 Alabio ducks were separated into four groups: domestic chicken and Alabio duck treatment groups, where each was infected by NDV velogenic ducks/Aceh Besar_IND/2013/eoAC080721 in 106ELD50 dosage. The domestic chicken and Alabio duck control groups were each inoculated with Phosphate Buffer Saline. The infection route was intraorbital, 0.1 ml in volume. Symptoms were observed from day 1 until day 7 post-infection (PI). Necropsy was performed on days 1, 2, 3, 5, and 7 PI for organ collection. Symptoms observed were disorders in the respiratory, gastrointestinal, and nervous systems, followed by 100% mortality in domestic chickens. In Alabio ducks, there were only depression and slight lethargy. The lesion in domestic chicken day 1 appeared by the lungs, thymus, Fabricius bursa, spleen, and kidney. On day 3 PI, lesions were also found in the heart, proventriculus, duodenum, and cecal tonsil. On days 5 and 7 PI, the trachea and brain lesions were found. In Alabio ducks, lesions were found in the lungs, thymus, spleen, and proventriculus on day 1. Afterward, on day 3, light lesions were found in the heart. On day 5, lesions were found in the trachea and brain; finally, on day 7, light lesions were only found in the thymus, spleen, and brain. Immunopositive reaction NDV in domestic chicken was highest in the proventriculus, duodenum, cecal tonsil, and lymphoreticular organs. In the Alabio duck, it was highest in the duodenum and cecal tonsil. The caspase-3 percentage in domestic chicken increased on day 3 PI; in Alabio ducks, on day 2 PI. Clinical symptoms and pathological lesions were faster and more severe in domestic chickens. The immunopositive reaction NDV in domestic chicken continued to increase, while in Alabio ducks, it decreased until the last observation day. Apoptosis percentage increased earlier in the Alabio duck than in domestic chicken.

Effect of non-genetic factors on semen characteristics of Murrah bulls under tropical condition

The objective of this study was to assess the effect of non-genetic factors on semen characteristics of 22479ejaculates from 139 Murrah bulls during the period of 2017-2020analyzed under tropical conditions, maintained and recorded at Frozen Semen Bank (FSB), Bassi, Rajasthan Cooperative Dairy Federation, Jaipur, Rajasthan, India as well as in Information Network for Animal Productivity &amp; Health (INAPH) applicationmaintained at National Dairy Development Board (NDDB). Results were recorded as 2.94±0.03 mL for volume, 1234.72±13.51 million/mL for sperm concentration and 68.44±0.25 percentage for motility. Statistical analysis presented a significant difference for all semen parameters with studied non-genetic factors except semen collector for motility traits. Also, seasonal dynamics presented winter as the most suitable season for semen collection under tropical conditions. Season of ejaculates was observed as most influencing factors for all semen characteristics in Murrah bulls. Age of bull showed significantly increased values for all semen traits with the increasing age of bull except motility traits. Data analysis in this regard may be utilized to enhance the fertility rateby increasing the semen quality.

Sample preconcentration through airjet-induced liquid phase enrichment.

Sample preparation is essential for nucleic acid assays, affecting their sensitivity and reliability. However, this process often results in a significant loss or dilution of the analyte, which becomes a bottleneck that limits downstream assay performance, particularly for assays that accept a limited input sample volume. To overcome this challenge, we present an evaporative-based sample enrichment method that uses an airjet to concentrate analytes within a small, defined volume by reversing the coffee-ring effect. A small, concentrated sample can then be collected for analysis to increase the initial sample load. The effectiveness of the reported airjet enrichment was quantified using qPCR of λ-DNA, HeLa-S3 RNA, and heat-inactivated SARS-CoV-2 samples. Comparisons between airjet enrichment and conventional evaporative concentration methods demonstrated significant advantages of airjet enrichment, including the ability to concentrate a high percentage of analyte within a 1 μL volume. The enrichment method was then integrated and adapted for various fluid volumes commonly found in nucleic acid sample preparation procedures. Here, airjet enrichment reduced the overall Cq by an average of 9.27 cycles for each analyte, resulting in a 600-fold enrichment from the initial concentration. To perform selective enrichment and prevent salt-based interference in downstream analysis, PEG was added to reduce the co-enrichment of salt. In addition, a preliminary study was conducted to explore the integration of airjet enrichment into ELISA using rabbit IgG as a model antigen. These findings demonstrate how airjet enrichment can be easily integrated into existing laboratory protocols with minimal modification and significantly improve the performance of biosensors.

Method for hepatitis B virus DNA detecting in biological material at low viral load based on nested PCR with detection on three viral targets in real-time mode.

A method has been developed for HBV DNA finding in biological material at low viral load based on nested PCR with real-time detection of three viral targets. When developing the method, blood plasma samples were used from 128 CHB patients living in the regions of the Russian Federation and countries of Central Asia and 173 hemodialysis center patients living in the North-West Federal District. Analytical sensitivity was tested using the stepwise dilution method. HBV was detected by nested PCR. According to the method developed by us, at the first stage, the HBV DNA is amplified using at the first stage oligonucleotides complementary to the greatest similarity regions of the various HBV isolates genomes flanking the entire virus genome. At the second stage, when using the amplification product of the first stage as a template, PCR was performed using three pairs of oligonucleotides and the corresponding oligonucleotide fluorescently labeled probes to three virus genome regions (Core gene, S gene and X gene), as well as one pair of primers and the corresponding probe complementary to a human HPRT gene region. The method sensitivity for DNA extraction from plasma with a 100 μl volume was 10 IU/ml. Obtaining a threshold Ct cycle for only one fluorophore may indicate the presence of HBV DNA in the sample at a load of less than 10 IU/ml, HBV detection in this case is possible with a repeated PCR study of the corresponding sample with HBV DNA extraction from an increased plasma volume (200-1000 μl). The developed method makes it possible to identify the disease in various HBV subgenotypes and can be used to diagnose CHB in the population and risk groups, including those with the HBsAg-negative form of the disease.

Decline of Sperm Quality over the Last Two Decades in the South of Europe: A Retrospective Study in Infertile Patients.

Semen quality has a direct relation to male fertility. Whether sperm variables in humans have decreased over the last years is still uncertain, with some studies showing a decline and others reporting no changes. In this regard, previous research has suggested that lifestyle and environmental conditions may contribute to this variability, calling for regional studies. The present work is a retrospective, unicentric study that includes semen samples analyzed between 1997 and 2017 at the Parc Taulí Hospital (Barcelona metropolitan area). First, a multivariate analysis including the age as a confounding factor showed a statistically significant decrease in semen volume, pH, progressive motility, morphology and total motile sperm over time. Contrarily, no significant variation in sperm count or concentration was observed. Mean reductions per year were -0.02 mL for volume, -0.57% for progressively motile sperm and -0.72% for sperm with normal morphology. Interestingly, the average annual temperature registered by the Spanish Meteorology Agency negatively correlated to sperm morphology and sperm count (Rs = -0.642; p = 0.002 and Rs = -0.435; p = 0.049, respectively). In conclusion, the present study based on infertile patients from the Barcelona area found a decline in sperm motility and morphology, without effects on sperm count. Changes in temperature appeared to be associated to this decline, but further studies are needed to address the mechanisms linked to the observed variations.

Development of the Polyphenol Compounds from Plant Raw Materials by the Cell Micropropagation IN VITRO Method

One of the ways to design polyfunctional ingredients that determine the functional properties of food products are complex food additives consisting of valuable plant raw materials. The additives quality depends on the used raw materials adequacy, time and collection place, and the plant cultivation conditions. The traditional plantation method requires significant costs to obtain secondary metabolites determining physiological plant materials value. The research aim is to obtain polyphenolic compounds from phyto-raw materials by microclonal cell propagation under sterile laboratory conditions. The leaves and berries cells of cowberries and cranberries containing a significant number of polyphenols are the research objects. For example, cranberries growing in the south of the Tyumen region contain anthocyanins 97.8 mg/100 g and leukoanthocyanins 459.6 mg/100 g, and berries growing in the Arctic territories of the Yamalo-Nenets Autonomous Okrug contain 224.7 and 480.2 mg/100 g, respectively. At the initial stage, the researchers sterilized objects, instruments, and equipment. They determined the sterilization duration experimentally. The study revealed that with a longer sterilization duration, there was a change in color and an increase in cell toxicity, with a minimum duration, pathogenic microorganisms remained in the medium. After sterilization, a man cultured the cells in the light under fluorescent lamps with illumination of 100 mmol quanta/m²; photoperiod of 20 h per day; in nutrient media with an acidity of pH 5.2–5.4; 25 ml in volume; enriched with the auxins and cytokinins hormones. The researchers used Kundu phytohormones to regulate the secondary metabolites synthesis. They utilized Murashige Skoog and Anderson agar media to find the optimal nutrient medium in order to obtain the maximum increase in polyphenolic compounds in plant cells. A man developed the third agar media independently, considering the prospects of an individual approach to the more efficient producers growth.

Transaxillary hemithyroidectomy and parathyroidectomy: mastering the technique

Background. The relevance of endoscopic interventions on the thyroid and parathyroid glands is beyond doubt, and this explains the interest in this promising choice of surgical approach both among surgeons and patients who want to avoid a scar on the neck.Aim. To evaluate the first results of own endoscopic interventions in patients with diseases of the thyroid and parathyroid glands.Materials and methods. In the Research Institute of KCH N 1 named after prof. S.V. Ochapovsky from December 2020 to April 2022, 23 hemithyroidectomies and 8 paraadenomectomies were performed using endoscopic techniques for benign neoplasms of the thyroid and parathyroid glands. All patients in the preoperative period underwent ultrasound, TAPB + CI (for thyroid pathology), and the hormonal background was studied. Indications for endoscopic hemithyroidectomy in 18 cases were colloid goiter, in 5 cases — follicular adenoma. The indication for endoscopic paraadenomectomy in all cases was primary hyperparathyroidism. We used endoscopic three-port axillary-mammary gas access. Under the ETN, an incision was made along the outer edge of the pectoralis major muscle (m. pectoralis major). Carbon dioxide was injected into the pancreas using a Verish needle. A 5 mm port was inserted along the edge of the areola. Another 10 mm trocar was installed at the attachment points (m. pectoralis major).Results. The average operation time was 190.4 minutes for endoscopic hemithyroidectomy and 78.6 minutes for endoscopic paraadenomectomy. With the increase in the number of performed operations, there was a decrease in the duration of surgical interventions. In two cases in patients after hemithyroidectomy, seromas up to 20 ml in volume were noted in the postoperative period, which required puncture interventions (once).Conclusion. Endoscopic interventions on the thyroid and parathyroid glands from axillary access are a safe method of surgical intervention, and the cosmetic result is an indisputable advantage of this method.