Heterogenous pattern of the inflammasome-related markers in the critically ill COVID-19 patients at the ICU admission.

Purpose: Development of targeted biological therapies to COVID-19 requires reliable biomarkers that could help to indicate the responding patients. Hyperactivation of the inflammasome by SARS-CoV2 virus is hypothesized to contribute to severe course of the COVID-19 disease. Therefore, we aimed to evaluate the prognostic value of several inflammasome-related cytokines and proteins at the admission to the intensive care unit. Patients and methods: Plasma samples were obtained from 45 critically-COVID-19 patients and from 10 patients with severe craniocerebral traumatic brain injury (TBI) at the admission to the ICU. The concentration of IL-1𝛼, IL-1𝛽, IL-18, IL-1RA, galectin-1, ASC, LDH, ferritin and gasdermin D were analyzed. A novel cell-free caspase-1 plasma assay was developed by inhibitor-based immunoprecipitation followed by Western Blot. Demographic and clinical characteristics were recorded. Results: The inflammasome-related biomarkers were in similar concentration in COVID-19 and TBI patients except for galectin-1 being lower in the former. None of the tested markers was related to the outcome, length of stay or development of secondary infections. Patients with SOFA score of >9 at admission who were at high risk of death had significantly higher galectin-1 but lower IL-1RA in comparison to low-risk patients. Weak but significant correlations were observed for IL-1𝛼 and IL-1𝛽 and platelets and also for ferritin and INR. Activated caspase-1 p35 was detectable in 12/22 COVID-19 patients and was related with higher fibrinogen and lower D-dimers. It was also significantly higher in patients with SOFA>9. Conclusion: Our results indicate that the activation of the inflammasome in critically ill COVID-19 patients is a heterogenous process and is not directly related with outcome. Therefore, potential interventions aimed at this pathway in this group of patients can be limited and should be biomarker-guided.

Effect of cryoprotectant on the motility, viability, fertilization, and DNA integrity of naleh fish Barbonymus sp. (Cyprinidae) sperm

Naleh fish Barbonymus sp. is a commercial freshwater fish, which is indigenous to Aceh, Indonesia. The population of this species has declined over the years as a result of habitat perturbations and overfishing. Hence, the crucial need to develop a cryopreservation method to support breeding programs. This involved the use of a cryoprotectant as an important component. The objective of this study, therefore, was to explore the best cryoprotectant for naleh fish spermatozoa, and a total of five types were tested. These include the DMSO, Methanol, Ethanol, Glycerol, and Ethylene Glycol at a similar concentration of 10%, which were individually combined with 15% egg yolk, and every treatment was performed in three replications. Conversely, Ringer’s solution was adopted as an extender, and the sperm was cryopreserved in liquid nitrogen for 15 days. The results showed significant influence on sperm motility and viability, as well as egg fertility of naleh fish (P <0.05), although the DMSO provided the best outcome, compared to others at 47.17%, 50.13%, and 45.67%, respectively. Furthermore, DNA fragmentation had not occurred in the fresh and cryopreserved sperm samples, indicating the protective effect of tested cryoprotectants. It is concluded that the 10% DMSO and 15% egg yolk is the best cryoprotectant for naleh fish spermatozoa.

Native α-Synuclein, 3-Nitrotyrosine Proteins, and Patterns of Nitro-α-Synuclein-Immunoreactive Inclusions in Saliva and Submandibulary Gland in Parkinson’s Disease

Background. Salivary α-synuclein (aSyn) and its nitrated form, or 3-nitrotyrosine-α-synuclein (3-NT-αSyn), hold promise as biomarkers for idiopathic Parkinson’s disease (IPD). Nitrative stress that is characterized by an excess of 3-nitrotyrosine proteins (3-NT-proteins) has been proposed as a pathogenic mechanism in IPD. The objective is to study the pathological role of native αSyn, 3-NT-αSyn, and 3-NT-proteins in the saliva and submandibulary glands of patients with IPD. Methods. The salivary and serum αSyn and 3-NT-proteins concentration is evaluated with ELISA in patients and controls. Correlations of αSyn and 3-NT-proteins content with clinical features of the disease are examined. Immunohistochemical 3-NT-αSyn expression in submandibulary gland sections is analyzed. Results. (a) Salivary concentration and saliva/serum ratios of native αSyn and 3-NT-proteins are similar in patients and controls; (b) salivary αSyn and 3-NT-proteins do not correlate with any clinical feature; and (c) three patterns of 3-NT-αSyn-positive inclusions are observed on histological sections: rounded “Lewy-type” aggregates of 10–25 µm in diameter, coarse deposits with varied morphology, and spheroid inclusions or bodies of 3–5 µm in diameter. “Lewy-type” and coarse inclusions are observed in the interlobular connective tissue of the gland, and small-sized bodies are located within the cytoplasm of duct cells. “Lewy-type” inclusions are only observed in patients, and the remaining patterns of inclusions are observed in both the patients and controls. Conclusions. The patients’ saliva presents a similar concentration of native αSyn and 3-nitrotyrosine-proteins than that of the controls, and no correlations with clinical features are found. These findings preclude the utility of native αSyn in the saliva as a biomarker, and they indicate the absence of nitrative stress in the saliva and serum of patients. As regards nitrated αSyn, “Lewy-type” inclusions expressing 3-NT-αSyn are observed in the patients, not the controls—a novel finding that suggests that a biopsy of the submandibulary gland, if proven safe, could be a useful technique for diagnosing IPD. Finally, to our knowledge, this is also the first description of 3-NT-αSyn-immunoreactive intracytoplasmic bodies in cells that are located outside the nervous system. These intracytoplasmic bodies are present in duct cells of submandibulary gland sections from all subjects regardless of their pathology, and they can represent an aging or involutional change. Further immunostaining studies with different antibodies and larger samples are needed to validate the data.

Stimulus Control of Odorant Concentration: Pilot Study of Generalization and Discrimination of Odor Concentration in Canines

Despite dogs’ widespread use as detection systems, little is known about how dogs generalize to variations of an odorant’s concentration. Further, it is unclear whether dogs can be trained to discriminate between similar concentration variations of an odorant. Four dogs were trained to an odorant (0.01 air dilution of isoamyl acetate) in an air-dilution olfactometer, and we assessed spontaneous generalization to a range of concentrations lower than the training stimulus (Generalization Test 1). Dogs generalized to odors within a 10-fold range of the training odorant. Next, we conducted discrimination training to suppress responses to concentrations lower than a concentration dogs showed initial responding towards in Generalization Test 1 (0.0025 air dilution). Dogs successfully discriminated between 0.0025 and 0.01, exceeding 90% accuracy. However, when a second generalization test was conducted (Generalization Test 2), responding at the 0.0025 concentration immediately recovered and was no different than in Generalization Test 1. Dogs were then tested in another generalization test (Compound Discrimination and Generalization) in which generalization probes were embedded within discrimination trials, and dogs showed suppression of responding to the 0.0025 concentration and lower concentrations in this preparation. These data suggest dogs show limited spontaneous generalization across odor concentration and that dogs can be trained to discriminate between similar concentrations of the same odorant. Stimulus control, however, may depend on the negative stimulus, suggesting olfactory concentration generalization may depend on relative stimulus control. These results highlight the importance of considering odor concentration as a dimension for generalization in canine olfactory research.

Correction of energometabolic disorders atsignificant iodine deficiency in calves

The paper is devoted to the development and use of theiodomethaboliccomplex based on iodine-iodide with polyvinyl alcohol (iodinol) and succinic acid.It was found that the inclusion of succinic acid or sodium succinate in a similar concentration in the composition of iodinol allowed not only qualitatively improving the pharmacological properties of iodinol, but also obtaining an energometabolic composition suitable for parenteral administration.The results of using the iodomethabolic compositionrevealed that with the parenteral administration to calves with clinically pronounced iodine deficiency, it provides effective normalization of pathological biochemical processes in the body of calves (metabolic effect) and pronounced activation of energy metabolism (energy effect), and in general, an energometabolic effect that ensures clinical recovery of animals from iodine deficiency.

Serological Cytokine and chemokine profile in pregnant women with COVID19 in Mexico City

On January 30th, 2020, the WHO declared the outbreak of COVID19, a disease due to the new coronavirus called SARS-CoV-2. Certain comorbidities, symptoms and signs are characteristic of COVID19 in the general population and in pregnant women. However, pregnant women are considered as a high-risk group for COVID19. To know about the frequency of comorbidities, signs and symptoms, the presence of lymphopenia, antibodies response to SARS-CoV2 and cytokine and chemokine serum concentration, six pregnant women with COVID19 were studied at the moment of admission. The lower concentration of CCL17 was detected in the Pregnant COVID19 group, similar concentration of IL-6 was also detected in non-pregnant and pregnant COVID19 patients. Our result show that pregnant and non-pregnant women with COVID19 has similar cytokine profile.

Development of Head Space Sorptive Extraction Method for the Determination of Volatile Compounds in Beer and Comparison with Stir Bar Sorptive Extraction

A headspace sorptive extraction method coupled with gas chromatography–mass spectrometry (HSSE–GC–MS) was developed for the determination of 37 volatile compounds in beer. After optimization of the extraction conditions, the best conditions for the analysis were stirring at 1000 rpm for 180 min, using an 8-mL sample with 25% NaCl. The analytical method provided excellent linearity values (R2 > 0.99) for the calibration of all the compounds studied, with the detection and quantification limits obtained being low enough for the determination of the compounds in the beers studied. When studying the repeatability of the method, it proved to be quite accurate, since RSD% values lower than 20% were obtained for all the compounds. On the other hand, the recovery study was successfully concluded, resulting in acceptable values for most of the compounds (80–120%). The optimised method was successfully applied to real beer samples of different types (ale, lager, stout and wheat). Finally, an analytical comparison of the optimised HSSE method, with a previously developed and validated stir bar sorptive extraction (SBSE) method was performed, obtaining similar concentration values by both methods for most compounds.

On the Analogy between Electrolytes and Ion-Generating Nanomaterials in Liquid Crystals

Nanomaterials in liquid crystals are a hot topic of contemporary liquid crystal research. An understanding of the possible effects of nanodopants on the properties of liquid crystals is critical for the development of novel mesogenic materials with improved functionalities. This paper focuses on the electrical behavior of contaminated nanoparticles in liquid crystals. More specifically, an analogy between electrolytes and ion-generating nanomaterials in liquid crystals is established. The physical consequences of this analogy are analyzed. Under comparable conditions, the number of ions generated by nanomaterials in liquid crystals can be substantially greater than the number of ions generated by electrolytes of similar concentration.

Research in the field of analysis of the soft dosage form «Metroterbinazol» used in medical practice

Aim. Development of a method of analysis of metronidazole and terbinafine in ointment «Metroterbinazol» using spectrophotometry. Materials and methods. A soft dosage form containing 0.1 g of metronidazole, terbinafine and 9.8 g of Tizol gel was used as the object of research. Pharmacopoeia reagents and the method of spectrophotometry were used for experimental work. Measurements were carried out on the SF-2000 spectrophotometer in the near ultraviolet region (λ = 210-340 nm). Results. To achieve this goal, we studied the electronic absorption spectra and optical characteristics of metronidazole and terbinafine in the substance, ethanol solution and in the presence of Tizol gel. The curve of ε (terb) – ε (met) to the wavelength observed a maximum at a wavelength of 223 nm, which corresponds to the maximum of absorption of terbinafine and is close to a less pronounced first absorption maximum of metronidazole (226 nm), also pronounced minimum at a wavelength of 312 nm, which is similar to the second maximum of absorption of metronidazole. The results obtained suggest that the optimal wavelengths are 223 nm and 312 nm. On an artificial mixture with a similar concentration of ingredients according to the prescription, testing was conducted for the analysis of drugs in the joint presence. As a result of experiments, it was found that the mass of metronidazole, found by the Firordt method and the simplified system of equations, is in the limit of 0.0959-0.1053 g, terbinafine – 0.1072-0.1110 g, which corresponds to the permissible deviations. Based on the research, a method for the spectrophotometric analysis of terbinafine and metronidazole in «Metroterbinazole» ointment has been developed that allows quantifying them within the permissible deviations provided for in the regulatory documentation. Conclusion. A method of spectrophotometric quantitative determination of terbinafine and metronidazole is proposed, which allows analyzing medicinal products with a relative error of ± 1.49-1.50% and establishing the quality of manufacturing of «Metroterbinazole» ointment within acceptable standards of deviations.