Coated Microneedle System for Delivery of Clotrimazole in Deep-Skin Mycoses.

Mycoses of the skin are infectious diseases caused by fungal microorganisms that are generally treated with topical agents. However, such therapy is often ineffective and has to be supported by oral use of active substances, which, in turn, can cause many side effects. A good alternative for the treatment of deep-skin mycoses seems to be microneedles (MNs). The aim of this research was to fabricate and evaluate the properties of innovative MNs coated with a hydrogel as potential carriers for clotrimazole (CLO) in the treatment of deep fungal skin infections. A 3D printing technique using a photo-curable resin was employed to produce MNs, which were coated with hydrogels using a dip-coating method. Hydrogels were prepared with carbopol EZ-3 Polymer (Lubrizol) in addition to glycerol and triisopropanolamine. Clotrimazole was introduced into the gel as the solution in ethanol or was suspended. In the first step of the investigation, a texture analysis of hydrogels was prepared with a texture analyzer, and the drug release studies were conducted with the use of automatic Franz diffusion cells. Next, the release profiles of CLO for coated MNs were checked. The last part of the investigation was the evaluation of the antifungal activity of the prepared systems, and the inhibition of the growth of Candida albicans was checked with the diffusion and suspended-plate methods. The texture profile analysis (TPA) for the tested hydrogels showed that the addition of ethanol significantly affects the following studied parameters: hardness, adhesiveness and gumminess, causing a decrease in their values. On the other hand, for the gels with suspended CLO, better spreadability was seen compared to gels with dissolved CLO. The presence of the active substance did not significantly affect the values of the tested parameters. In the dissolution study, the results showed that higher amounts of CLO were released for MNs coated with a hydrogel containing dissolved CLO. Also, microbiological tests proved its efficacy against fungal cultures. Qualitative tests carried out using the diffusion method showed that circular zones of inhibition of fungal growth on the plate were obtained, confirming the hypothesis of effectiveness. The suspension-plate technique confirmed the inhibitory effect of applied CLO on the growth of Candida albicans. From the analysis of the data, the MNs coated with CLO dissolved in hydrogel showed better antifungal activity. All received results seem to be helpful in developing further studies for MNs as carriers of antifungal substances.

A continuous epistasis model for predicting growth rate given combinatorial variation in gene expression and environment.

Quantifying and predicting growth rate phenotype given variation in gene expression and environment is complicated by epistatic interactions and the vast combinatorial space of possible perturbations. We developed an approach for mapping expression-growth rate landscapes that integrates sparsely sampled experimental measurements with an interpretable machine learning model. We used mismatch CRISPRi across pairs and triples of genes to create over 8,000 titrated changes in E. coli gene expression under varied environmental contexts, exploring epistasis in up to 22 distinct environments. Our results show that a pairwise model previously used to describe drug interactions well-described these data. The model yielded interpretable parameters related to pathway architecture and generalized to predict the combined effect of up to four perturbations when trained solely on pairwise perturbation data. We anticipate this approach will be broadly applicable in optimizing bacterial growth conditions, generating pharmacogenomic models, and understanding the fundamental constraints on bacterial gene expression. A record of this paper's transparent peer review process is included in the supplemental information.

Actual status and future directions of cardiac telerehabilitation.

Telerehabilitation (TR) was developed to achieve the same results as would be achieved by the standard rehabilitation process and to overcome potential geographical barriers and staff deficiencies. This is especially relevant in periodic crisis situations, including the recent COVID-19 pandemic. Proper execution of TR strategy requires both well-educated staff and dedicated equipment. Various studies have shown that TR may have similar effects to traditional rehabilitation in terms of clinical outcomes and may also reduce total healthcare costs per participant, including rehospitalization costs. However, as with any method, TR has its advantages and disadvantages, including a lack of direct contact or prerequisite, rudimentary ability of the patients to handle mobile devices, among other competencies. Herein, is a discussion of the current status of TR, focusing primarily on cardiac TR, describing some technical/organizational and legal aspects, highlighting the indications, examining cost-effectiveness, as well as outlining possible future directions.

Rheological and textural analysis as tools for investigation of drug-polymer and polymer-polymer interactions on the example of low-acyl gellan gum and mesalazine.

PURPOSE: In the performed study, the rheological and textural parameters of gellan-based hydrogels were investigated and their dependence on three factors was taken into consideration: (i) The presence of the model drug, (ii) The presence and type of the ionic crosslinking agent, and (iii) the composition of the polymer network. The objective was to compare two analytical methods, regarded as complementary, and define to what extent the obtained results correlate with each other. METHODS: The hydrogels contained low-acyl gellan gum or its mixtures with hydroxyethyl cellulose or κ-carrageenan. CaCl2 and MgCl2 were used as gelling agents. Mesalazine was used as a model drug. The rheological analysis included oscillatory stress and frequency sweeping. The texture profile analysis was performed to calculate texture parameters. RESULTS: Placebo gels without the addition of gelling agents had the weakest structure. The drug had the strongest ability to increase the stiffness of the polymer network. The weakest structure revealed the placebo samples without the addition of gelling agents. Texture analysis revealed no significant influence of the drug on the strength of the gels, while rheological measurements indicated clear differences. CONCLUSIONS: It can be concluded that in the case of some parameters methods correlate, that is, the effect related to gelling ions. However, the rheological analysis seems to be more precise and sensitive to some changes in the mechanical properties of the gels.

Iris melanoma with ciliary body infiltration presenting as a presumed chronic unilateral inflammatory glaucoma. The challenges of iris tumor biopsy assessment.

Uveal melanoma is the most common primary malignancy of the eye in adults. It may involve the choroid and ciliary body, and in only 2-3% of cases it involves the iris. We present a case of a 56-year-old patient with a 6-year history of unilateral, inflammatory, refractory glaucoma of the right eye. Due to acquired heterochromia and heterogeneous thickness of the iris, iris melanoma was suspected, but the incisional biopsy did not confirm the diagnosis. In the next months, the lesion enlarged and the eye globe was enucleated. Histopathological examination revealed an iridociliary melanoma with annular growth pattern.

Effectiveness of Resistance Training with the Use of a Suspension System in Patients after Myocardial Infarction.

The aim of the study was to assess the effects of resistance training with the use of a suspension system on exercise tolerance, evaluated through an exercise test, and the changes in selected echocardiographic parameters of patients after myocardial infarction. The study involved 44 males. The subjects were divided into two groups: Standard (20) and Suspension system (24). All the subjects had undergone an angioplasty with stent implantation. The standard and suspension system groups carried out a 24-day improvement program comprising 22 training units. Each session consisted of endurance, general stamina and resistance training. Instead of resistance training, the experimental group made multijoint exercises with a suspension system. Statistically significant changes in both groups were observed in the parameters of the echocardiographic exercise test, such as test duration (p = 0.000), distance covered (p = 0.000), MET (p = 0.000), VO2max (p = 0.000) and SBPrest (p = 0.013). Additionally, SBPmax in the suspension system group improved (p = 0.035). The echocardiographic test revealed significant improvement of Left Ventricular Ejection Fraction in both groups (SP group p = 0.001, standard group p = 0.005). The lipid profile test in the SP group revealed statistically significant improvement of TC (p = 0.003), HDL (p = 0.000) and LDL (p = 0.005). Training with the suspension system had a positive effect on the change of exercise tolerance level, left ventricular function and blood lipid profile.

Effects of Hatha Yoga on Cardiac Hemodynamic Parameters and Physical Capacity in Cardiac Rehabilitation Patients.

PURPOSE: The purpose of the present study was to assess the effect of hatha yoga training that was added to the standard cardiac rehabilitation (CR) program on the cardiac hemodynamic parameters and physical capacity of patients with ST-elevation myocardial infarction (STEMI). METHODS: The study included 70 male patients aged 45-65 yr with STEMI who were treated by angioplasty. Patients were randomized to standard CR (control group) versus standard CR plus hatha yoga (experimental group). The training program lasted for a total of 24 d for each patient, with day 1 and day 24 used for medical examinations (electrocardiogram, spiroergometric submaximal treadmill test, and echocardiography). The remaining 22 d consisted of the actual training. RESULTS: After the CR program the spiroergometric stress test parameters and left ventricular ejection fraction (LVEF) improved in both the experimental and control groups. The most notable changes in echocardiography parameters and physical capacity were in the experimental group. The results showed significant main effect over time, a time-versus-group interaction in LVEF, the duration of the test, and peak oxygen uptake, and a time-versus-group interaction in metabolic equivalents (METs). We also noted the improvement of left ventricular end-diastolic diameter, left ventricular end-systolic diameter, and heart rate over time. CONCLUSION: The results revealed better effectiveness in the CR program with a modified hatha yoga training program. Hatha yoga training could be recommended as an adjunct to standard CR.

E-Beam Irradiation and Ozonation as an Alternative to the Sulphuric Method of Wine Preservation.

Potassium metabisulphite is usually used for microbial stabilization in the process of vinification and wine preservation, but it is considered to be allergenic. The objective of the present study was to assess the efficiency of ozonation and ionizing radiation as alternatives to wine sulphurization. The efficiency of yeast removal and the retention of the chemical quality of wine were evaluated. Wine was subjected to 60 min of ozonation, and radiation doses were set at 1-10 kGy. Moreover, a combination of ozonation and ionizing radiation treatment was used. The ozonation of wine did not produce the expected results. That is, it did not limit the number of yeast cells. From the sixth minute, a significant deterioration in the taste and the color of the wine was found. Ionizing radiation at a dose of 1 kGy reduced the yeast count by 95.5%, and a reduction of 99.9% was seen after the application of 2.5 kGy. Moreover, these doses did not have a significant effect on the organoleptic properties or the chemical composition of wine. The total amount of polyphenols reduced from the maximum of 1127.15 to 1023.73 mg at the dose of 5 kGy. Radiation is widely used to preserve food products. Its use for finished wine preservation may be an alternative to sulphurization.

Stimulation of the activity of a novel tannase produced in white-rot fungi Phellinus pini, Fomes fomentarius, and Tyromyces pubescens by medium supplementation.

In recent years, tannase has gained increasing interest mainly because of its potential applications. One of the most important functions of tannic acid (TA) hydrolase is the release of gallic acid (GA) from complex tannins. The aim of the study was to determine the dynamic changes in tannase activity depending on the carbon source in the culture medium. An extracellular and intracellular tannase activity analysis was carried out with the use of spectrophotometric analysis and confirmed by capillary electrophoresis in cultures of white-rot fungi: Phellinus pini, Fomes fomentarius, and Tyromyces pubescens. The inducible potential of TA and rapeseed meal on the activity of tannin acyl hydrolase was confirmed during 14 days of culturing. Different effects of the tested compounds on stimulation of tannase activity in selected fungal strains have been demonstrated. We concluded that rapeseed meal was the best inducer of tannase activity in the case of P. pini. However, the highest concentrations of GA were observed after stimulation by the TA in the cultures of F. fomentarius and T. pubescens.

[Quality of life and physical activity of patients with inflammatory bowel diseases]. / Aktywnosc fizyczna i jakosc zycia pacjentów z nieswoistymi zapaleniami jelit.

AIM OF THE STUDY: Estimation of the quality of life of patients with inflammatory bowel diseases and comparison these results with control group. MATERIAL AND METHODS: A group of 16 patients with inflammatory bowel diseases and 13 healthy persons as a control group. In orderto estimate the quality of life, polish version of Inflammatory Bowel Disease Questionnaire (IBDQ) was used. The Second questionnaire that was used is WHOQOL-BREF (The World Health Organization Quality of Life). To assess the level of physical activity was applied the Minnesota Leisure Time Physical Activity Questionnaire (MLTPAQ). All the results have been correlated to answer the question if there is any relationship between the quality of life and physical activity and if level of these parameters is different in control group. RESULTS: In the group of patients the lowest level of functioning were stated on bowel ailments field and emotional field. The most important roles in patients life are social relationship and sanity. The higher level of the caloric consumption was stated in the group of patients with inflammatory bowel diseases in comparison to control group. There was no significant difference in caloric consumption related with the recreation and house works. Because of the fact that all of the patients were in the remission period, it was impossible to verify if there is any relationship between quality of life and the stage of disease. CONCLUSIONS: The results of researches have not proved relationship between the level of physical activity and the quality of life among the patients as well as in the control group.

Mapping the N-glycome of human von Willebrand factor.

vWF (von Willebrand factor) is a key component for maintenance of normal haemostasis, acting as the carrier protein of the coagulant Factor VIII and mediating platelet adhesion at sites of vascular injury. There is ample evidence that vWF glycan moieties are crucial determinants of its expression and function. Of particular clinical interest, ABH antigens influence vWF plasma levels according to the blood group of individuals, although the molecular mechanism underlying this phenomenon remains incompletely understood. The present paper reports analyses of the human plasma vWF N-glycan population using advanced MS. Glycomics analyses revealed approximately 100 distinct N-glycan compositions and identified a variety of structural features, including lactosaminic extensions, ABH antigens and sulfated antennae, as well as bisecting and terminal GlcNAc residues. We estimate that some 300 N-glycan structures are carried by human vWF. Glycoproteomics analyses mapped ten of the consensus sites known to carry N-glycans. Glycan populations were found to be distinct, although many structural features were shared across all sites. Notably, the H antigen is not restricted to particular N-glycosylation sites. Also, the Asn(2635) site, previously designated as unoccupied, was found to be highly glycosylated. The delineation of such varied glycan populations in conjunction with current models explaining vWF activity will facilitate research aimed at providing a better understanding of the influence of glycosylation on vWF function.

O-linked glycosylation of von Willebrand factor modulates the interaction with platelet receptor glycoprotein Ib under static and shear stress conditions.

We have examined the effect of the O-linked glycan (OLG) structures of VWF on its interaction with the platelet receptor glycoprotein Ibα. The 10 OLGs were mutated individually and as clusters (Clus) on either and both sides of the A1 domain: Clus1 (N-terminal side), Clus2 (C-terminal side), and double cluster (DC), in both full-length-VWF and in a VWF construct spanning D' to A3 domains. Mutations did not alter VWF secretion by HEK293T cells, multimeric structure, or static collagen binding. The T1255A, Clus1, and DC variants caused increased ristocetin-mediated GPIbα binding to VWF. Platelet translocation rate on OLG mutants was increased because of reduced numbers of GPIbα binding sites but without effect on bond lifetime. In contrast, OLG mutants mediated increased platelet capture on collagen under high shear stress that was associated with increased adhesion of these variants to the collagen under flow. These findings suggest that removal of OLGs increases the flexibility of the hinge linker region between the D3 and A1 domain, facilitating VWF unfolding by shear stress, thereby enhancing its ability to bind collagen and capture platelets. These data demonstrate an important functional role of VWF OLGs under shear stress conditions.

Specific N-linked glycosylation sites modulate synthesis and secretion of von Willebrand factor.

We examined the role that N-linked glycans play in the synthesis and expression of von Willebrand Factor (VWF). Blocking the addition of N-linked glycans (NLGs) or inhibiting initial glycan processing prevented secretion of VWF. To determine whether specific glycosylation sites were important, the 16 VWF N-linked glycosylation sites were mutated followed by expression in HEK293T cells. Four NLG mutants affected VWF expression: N99Q (D1 domain), N857Q (D' domain), N2400Q (B1 domain), and N2790Q (CK domain) either abolished or reduced secretion of VWF and this was confirmed by metabolic labeling. Multimer analysis of mutant N2790Q cell lysate revealed an increase in VWF monomers, which was also observed when the isolated CK domain was expressed with N2790 mutated. Immunofluorescence microscopy showed that mutants N99Q, N857Q, and N2790Q were primarily retained within the ER, producing only few pseudo Weibel-Palade bodies over longer time periods compared with wtVWF. All the variants also showed an increase in free thiol reactivity. This was greatest with N857Q and D4-C2 NLG mutants, which had approximately 6-fold and 3- to 4-fold more free thiol reactivity than wtVWF. These data provide further evidence of the critical role that individual N-linked glycans play in determining VWF synthesis and expression.

The obstetrical history in patients with Pfannenstiel scar endometriomas--an analysis of 81 patients.

INTRODUCTION: The participation of immune tolerance during pregnancy was suggested to be an important factor predisposing to the implantation of decidual cells after cesarean section in Pfannenstiel scar. Delivery at term is related to the termination of immune tolerance to fetal antigens that is maintained throughout pregnancy. Substantial proportion of cesarean section deliveries is performed before the onset of true term labor. The aim of this study was to analyze the clinical symptoms of spontaneous beginning of labor in pregnant women in whom cesarean sections were performed and in whom Pfannenstiel scar endometriomas were observed during follow-up. MATERIALS AND METHODS: We have retrospectively analyzed 81 patients following the surgical removal of scar endometrioma after cesarean section. Obstetrical histories of cesarean sections in the number of 5,370 preceding the occurrence of the scar endometrioma were analyzed. These data were collected in six different Gynecological and Obstetrical wards in Malopolska Province in Poland. Analysis of data was started by the retrospective evaluation of regular uterine contractions, uterine cervix ripening before cesarean section and the indications for surgery. RESULTS: In 67 women from the group of 81 patients cesarean sections were performed with unripe uterine cervix and without the presence of regular uterine contractions. Elective indications for cesarean sections were predominant in this group of women. The relative risk of scar endometriomas occurrence following cesarean sections performed before onset of labor in comparison to cesarean sections following spontaneous onset of labor was statistically significantly higher [RR = 2.16, 95% CI = 1.21-3.83; OR = 2.18, 95% CI = 1.22-3.89]. CONCLUSIONS: Cesarean section performed before spontaneous onset of labor may increase substantially the risk of occurrence of scar endometriomas.