Effect of Copper on the Function of Isolated Porcine Kidneys Stored Using Simple Hypothermia.

Renal ischemia in the peri-transplant period causes a number of changes that adversely affect the initiation of normal vital functions in grafts after transplantation. To minimise the extent of ischemic damage, organs are stored in preservation fluid. The components of the fluid are supposed to ensure stabilisation of the cell cytoskeleton, protect against oxygen free radicals, reduce cell swelling, and ensure endothelial cell integrity. The aim of this study was to analyse the protective effect of Cu2+, as a component of Biolasol preservative fluid, in the prevention of nephron damage occurring during the graft storage period. Analyses of the effectiveness of copper in the presence of prolactin added to Biolasol fluid were also carried out. Forty isolated pig slaughter kidneys were used in the study, avoiding the use of laboratory animals. The kidneys were stored using simple hypothermia. After 2 h and 48 h of graft storage, selected biochemical indicators of renal function were determined in the collected perfusates. The addition of Cu2+ at a dose of 1 µg/L to the composition of Biolasol fluid was found to affect the generation of ischemic damage in the isolated pig kidney. The intensity of the occurrence of these processes is exacerbated by the presence of prolactin at a dose of 0.1 µg/L.

Effectiveness Assessment of a Modified Preservation Solution Containing Thyrotropin or Follitropin Based on Biochemical Analysis in Perfundates and Homogenates of Isolated Porcine Kidneys after Static Cold Storage.

In this paper, we assess the nephroprotective effects of thyrotropin and follitropin during ischaemia. The studies were performed in vitro in a model of isolated porcine kidneys stored in Biolasol (FZNP, Biochefa, Sosnowiec, Poland) and modified Biolasol (TSH: 1 µg/L; FSH 1 µg/L). We used the static cold storage method. The study was carried out based on 30 kidneys. The kidneys were placed in 500 mL of preservation solution chilled to 4 °C. The samples for biochemical tests were collected during the first kidney perfusion (after 2 h of storage) and during the second perfusion (after 48 h of storage). The results of ALT, AST, and LDH activities confirm the effectiveness of Biolasol + p-TSH in maintaining the structural integrity of renal cell membranes. Significantly reduced biochemical parameters of kidney function, i.e., creatinine and protein concentrations were also observed after 48 h storage. The protective effect of Biasol + p-TSH is most pronounced after 2 h of storage, suggesting a mild course of damage thereafter. A mild deterioration of renal function was observed after 48 h. The results of our analyses did not show any protective effect of Biolasol + p-FSH on the kidneys during ischaemia.

Biochemical Studies in Perfundates and Homogenates of Isolated Porcine Kidneys after Flushing with Zinc or Zinc-Prolactin Modified Preservation Solution Using a Static Cold Storage Technique.

Zinc is an effective anti-inflammatory and antioxidant trace element. The aim of this study was to analyse the protective effect of zinc and zinc-prolactin systems as additives of preservation solutions in the prevention of nephron damage caused during ischemia. The study used a model for storing isolated porcine kidneys in Biolasol®. The solution was modified with the addition of Zn at a dose of 1 µg/L and Zn: 1 µg/L with prolactin (PRL): 0.1 µg/L. After 2 h and 48 h of storage, the levels of alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, sodium, potassium, creatinine and total protein were determined. Zinc added to the Biolasol® composition at a dose of 1 µg/L showed minor effectiveness in the protection of nephrons. In turn, Zn2+ added to Biolasol + PRL (PRL: 0.1 µg/L) acted as a prolactin inhibitor. We do not recommend the addition of Zn(II) (1 µg/L) and Zn(II) (1 µg/L) + PRL (0.1 µg/L) to the Biolasol solution.

The Role of Hormones and Trophic Factors as Components of Preservation Solutions in Protection of Renal Function before Transplantation: A Review of the Literature.

Transplantation is currently a routine method for treating end-stage organ failure. In recent years, there has been some progress in the development of an optimal composition of organ preservation solutions, improving the vital functions of the organ and allowing to extend its storage period until implantation into the recipient. Optimizations are mostly based on commercial solutions, routinely used to store grafts intended for transplantation. The paper reviews hormones with a potential nephroprotective effect, which were used to modify the composition of renal perfusion and preservation solutions. Their effectiveness as ingredients of preservation solutions was analysed based on a literature review. Hormones and trophic factors are innovative preservation solution supplements. They have a pleiotropic effect and affect normal renal function. The expression of receptors for melatonin, prolactin, thyrotropin, corticotropin, prostaglandin E1 and trophic factors was confirmed in the kidneys, which suggests that they are a promising therapeutic target for renal IR (ischemia-reperfusion) injury. They can have anti-inflammatory, antioxidant and anti-apoptotic effects, limiting IR injury.

Therapeutic Potential of Selenium as a Component of Preservation Solutions for Kidney Transplantation.

Selenium has strong antioxidant properties and diverse effects on the immune system. The aim of the study was to analyse the protective effect of selenium as a component of a kidney preservation solution on the prevention of ischemia-reperfusion injury of nephrons. The solution was modified by the addition of Se (1 µg/L), prolactin (0.1 µg/L) and Se with prolactin (1 µg/L Se + 0.1 µg/L PRL). The study used a model for storing isolated porcine kidneys in Biolasol® (modified Biolasol®), which minimizes ischemia-reperfusion injury of grafts. The introduction of Se4+ ions at a dose of 1 µg/L into the Biolasol® preservation solution in the form of Na2SeO3 caused an increase in the activity/concentration of the analysed biochemical parameters: aspartate transaminase, alanine transaminase, urea and protein. This suggests an adverse effect of Se4+ on nephron function during ischemia-reperfusion. The best graft protection was obtained by using Biolasol® modified with the addition of selenium (IV) at a dose of 1 µg/L and prolactin at a concentration of 0.1 µg/L. We proposed the mechanism of prolactin action in the metabolic reduction of selenite (SO32-) during ischemia/reperfusion.

Development and Study of Semi-Solid Preparations Containing the Model Substance Corticotropin (ACTH): Convenience Application in Neurodegenerative Diseases.

Corticotropin (ACTH, previously an adrenocorticotropic hormone) is used in the diagnosis and treatment of pituitary gland disorders, adrenal cortex disorders, and other diseases, including autoimmune polymyositis, systemic lupus erythematosus, rheumatoid arthritis, Crohn's disease, and ulcerative colitis. So far, the ointment dosage form containing ACTH for use on the skin is unknown. Therefore, it seems appropriate to develop a semi-solid formulation with corticotropin. Emulsion ointments were prepared using an Unguator based on the cream base Lekobaza® containing corticotropin in different concentrations, and then the physical and chemical parameters of the ointment formulations, such as pH, spreadability, rheological properties, and texture analysis, were evaluated. In addition, a USP apparatus 2 with enhancer cells was utilized to study the in vitro drug release characteristics of the selected formulations. All the ointments obtained were characterized by good spreadability and viscosity. An analysis of the ointment texture was performed and the dependence of the tested parameters on the ACTH content in the ointment was demonstrated. Examination of the structure of the ointment showed that a high concentration of ACTH increases the hardness and adhesiveness of the ointment. In turn, it adversely affects the cohesiveness and elasticity of the ointments tested. The results of the release study showed that ACTH is released the fastest from the formulation with the lowest concentration, while the slowest from the ointment with the highest concentration of ACTH.

Influence of Concentration on Release and Permeation Process of Model Peptide Substance-Corticotropin-From Semisolid Formulations.

The transdermal route of administration of drug substances allows clinicians to obtain a therapeutic effect bypassing the gastrointestinal tract, where the active substance could be inactivated. The hormonal substance used in the study-corticotropin (ACTH)-shows systemic effects. Therefore, the study of the effect of the type of ointment base and drug concentration on the release rate and also permeation rate in in vivo simulated conditions may be a valuable source of information for clinical trials to effectively optimize corticotropin treatment. This goal was achieved by preparation ointment formulation selecting the appropriate ointment base and determining the effect of ACTH concentration on the release and permeation studies of the ACTH. Semi-solid preparations containing ACTH were prepared using Unguator CITO e/s. The release study of ACTH was tested using a modified USP apparatus 2 with Enhancer cells. The permeation study was conducted with vertical Franz cells. Rheograms of hydrogels were made with the use of a universal rotational rheometer. The dependence of the amount of released and permeated hormone on the ointment concentration was found. Based on the test of ACTH release from semi-solid formulations and evaluation of rheological parameters, it was found that glycerol ointment is the most favourable base for ACTH. The ACTH release and permeation process depends on both viscosity and ACTH concentration. The higher the hormone concentration, the higher the amount of released ACTH but it reduces the amount of ACTH penetrating through porcine skin.

Mood, cognitive structuring and medication adherence.

A study with a placebo was conducted. Healthy university students were given a placebo and were told to make one pill every day for a week. Participants were informed that the medicine improved mood. The extent to which they conformed to this instruction was treated as an index of compliance. Our results show that for women, but not for men, positive mood and cognitive structuring or negative mood and lack of cognitive structuring significantly predicted participants' compliance. A new model of medication adherence, based on the role of the patient's mood and cognitive structuring processes in decision making is presented in the paper.

Insulin-Loaded Chitosan-Cellulose-Derivative Hydrogels: In Vitro Permeation of Hormone through Strat-M® Membrane and Rheological and Textural Analysis.

This work is part of the current research trend to develop a hydrogel carrier of insulin to promote wound healing. Topically applied insulin promotes keratinocyte proliferation and migration, increases collagen synthesis, reduces inflammation and oxidative stress, and exhibits antimicrobial activity. The aim of this study was to design an insulin hydrogel matrix based on selected cellulose derivatives (methylcellulose, hydroxyethylcellulose, and hydroxypropylmethylcellulose) and chitosan. Rheological parameters of the formulations were evaluated using rotational rheometry and an oscillation test. Textural tests were performed. In vitro pharmaceutical insulin availability studies were carried out using the innovative Strat-M® membrane to imitate the skin barrier. It was found that the pharmaceutical formulation of insulin based on chitosan and methylcellulose showed an acceptable balance between rheological and textural parameters and ease of application. The API was released from the carrier in a prolonged manner, eliminating the need to apply the formulation several times per day. The developed hydrogel shows potential for use in clinical practice.

Study of the Effect of Phosvitin as a Potential Carrier on the Permeation Process of Somatotropin (STH) and Corticotropin (ACTH) from Biodegradable Polymers Used as Vehicles for STH and ACTH in Semi-Solid Formulations for Skin Application.

Phosvitin shows chelating abilities, an affinity for ACTH (corticotropin), growth factors, antioxidant properties, and acidic nature. An attempt was made to use this protein in hydrogels as a transporter of other protein substances: somatotropin (STH) and (ACTH). The aim of the study was to evaluate the effect of phosvitin on the permeation of ACTH and STH from semi-solid forms of the drug applied to the skin. Four hydrogel substrates were prepared using natural polymers: sodium alginate, methylcellulose, and starch. Based on the evaluation of physicochemical parameters, the hydrogel with the most favorable properties was selected and loaded with the active substances STH and ACTH, followed by the addition of phosvitin. A study of the permeation of STH and ACTH through the artificial cellulose membrane and through porcine skin was carried out without and with the addition of phosvitin. The effect of protein substances on rheological and textural parameters was studied. The evaluation of physicochemical parameters showed a favorable effect of STH and Phosvitin on the stability of the hydrogel with 4% methylcellulose and no effect of ACTH. All prepared formulations showed a reaction close to the natural pH of human skin. In the porcine skin permeation study, the addition of Phosvitin to the hydrogel with STH caused a slight increase in the amount of STH permeated and an increase in the time for STH to permeate porcine skin by 30 min. Phosvitin caused an increase in the amount of ACTH permeated through porcine skin almost twofold. Phosvitin may prove to be a promising permeation promoter for model protein-peptide substances when applied to the skin surface.

Resemblance and investment in children.

According to evolutionary explanations men hardly ever are absolutely certain about their biological fatherhood therefore they must seek various sources of information to subjectively establish whether they are the genetic fathers of the children they raise. Apicella and Marlowe (2004) showed that fathers who perceived greater similarity between their children and themselves were willing to invest more resources (e.g., time, money, care) in their offspring presumably because the perceived resemblance indicated to the fathers their genetic relatedness with their children. The present study extended the design of Apicella and Marlowe's original study and included both fathers and mothers as participants. Parents were recruited by a female confederate at the airport and at the railway station in Wroclaw (Poland). Multiple regression analyses showed that perceived resemblance predicted parental investment in the child for both men and women. The fact that mothers' declarations of investment in their children also depended on the perceived resemblance factor is not consistent with evolutionary formulations delineated by Apicella and Marlowe (2004; 2007). Future studies must resolve the issue of whether the resemblance-investment relation in fathers results from men relaying on child's resemblance to themselves as an indicator of their own biological paternity, or whether it results from the more parsimonious phenomenon that people in general are attracted more to other people who are similar to them.

Hydrogel Formulations for Topical Insulin Application: Preparation, Characterization and In Vitro Permeation across the Strat-M® Membrane.

Insulin has shown efficacy in the treatment of hard-to-heal wounds, which is mainly due to its role in regulating oxidative stress and inflammatory reactions. The aim of this study was to develop an insulin-hydrogel carrier based on Sepineo™ P 600 and Sepineo™ PHD 100 for application to lesional skin. Preformulation studies of the developed formulations were performed in terms of analysis of the pharmaceutical availability of insulin from the hydrogels through the Strat-M® membrane, and rheological and texture measurements. Insulin is released in a prolonged manner; after a time of 6.5 h, 4.01 IU/cm2 (53.36%) and 3.69 IU/cm2 (47.4%) of the hormone were released from the hydrogel based on Sepineo™ P 600 and Sepineo™ PHD 100, respectively. Rheological analysis showed that the hydrogels tested belong to non-Newtonian, shear-thinning systems with yield stress. The insulin-hydrogel based on Sepineo™ P 600 and Sepineo™ PHD 100 shows optimal application properties. The results obtained provide a basis for further preclinical and clinical studies.

Permeation of albumin through the skin depending on its concentration and the substrate used in simulated conditions in vivo.

OBJECTIVE: Many drugs applied to the skin with a systemic effect do not have a therapeutic effect, due to the barrier posed by the complex structure of the skin. To counteract this, absorption promoters are often added to the drug formulation. The use of albumin as an effective drug carrier is increasingly being addressed. Albumin, a natural, non-toxic polymer, can target drugs to specific cells and extend their biological half-life. This study was designed to trace the permeation of albumin after topical administration to the skin as a potential carrier of therapeutic substances. MATERIALS AND METHODS: Four dermal formulations based on different polymers were prepared: methyl cellulose, sodium alginate, hypromellose and chitosan with methyl cellulose, obtaining final concentrations of albumin of 2%, 1.5% and 1%. The permeation of albumin through the skin was examined under simulated in vivo conditions. RESULTS: Most albumin permeated from the methylcellulose-based hydrogel. Depending on the concentration of albumin, permeation profiles were plotted and permeation rate constant and AUC(0-24 h) were calculated. CONCLUSION: Methylcellulose was the optimal polymer for albumin release, whereas hypromellose was the least favorable. The concentration of albumin influences the amount and rate of permeation of this protein. The optimal concentration was 10 mg/g, from which the most albumin penetrated and the fastest. Human skin appeared to be more permeable to albumin than pig skin. However, the similar permeation profile through both membranes successfully allows the use of pig skin to track and evaluate the permeation of therapeutic substances with systemic effects.

Effect of Manganese on the Efficacy of Preservative Solution in Protecting Isolated Porcine Kidneys.

BACKGROUND: Manganese is a bioelement essential for the proper functioning of the body. Mn2+ is a cofactor for many enzymes, including manganese superoxide dismutase. Together with the enzymes, it participates in cell defense against reactive oxygen species, immune response, and energy production in mitochondria. The aim of this study was to evaluate the effect of manganese added to Biolasol solution on the storage efficiency of isolated pig kidneys. METHODS: Twenty kidneys from Polish Great White pigs were used in the study. Biolasol solution was modified with the addition of Mn2+ at a concentration of 1 µg/L. Grafts were assigned to 2 groups: A/Biolasol (n = 10) and B/Biolasol + Mn2+ (n = 10). The kidneys were washed with the developed solutions, then stored using the static method for 48h under hypothermia (4°C) and washed again. Markers of normal renal function were analyzed in the collected perfusates-aspartate aminotransferase (AST), alanine aminotransferase (ALT), protein, creatinine, urea, and electrolytes. RESULTS: After 48 hours and 30 minutes of renal preservation, activities/concentrations of biochemical parameters in the Biolasol + Mn2+ group increased (ALT: by 33%; AST: by 86%; urea: by 18%; creatinine: 3.25-fold [P < .05]) vs Biolasol. K+ ion concentration was 33% lower in the Biolasol group after 48 hours and 30 minutes (P < .05) vs group B. The presence of manganese in the composition of Biolasol solution influenced the maintenance of optimal Na+ ion concentration. CONCLUSIONS: Manganese added to the Biolasol solution formulation at a dose of 1 µg/L showed low efficacy in protecting nephrons. We do not recommend adding manganese to the preservative solution formulation.

Media intervention program for reducing unrealistic optimism bias: The link between unrealistic optimism, well-being, and health.

Unrealistic optimism is the tendency to perceive oneself as safer than others in situations that equally threaten everybody. By reducing fear, this bias boosts one's well-being; however, it is also a deterrent to one's health. Three experiments were run in a mixed-design on 1831 participants to eliminate unrealistic optimism (measured by two items-probability of COVID-19 infection for oneself and for others; within-subjects) toward the probability of COVID-19 infection via articles/videos. A between-subject factor was created by manipulation. Ostensibly, daily newspaper articles describing other people diligently following medical recommendations (experiment 1) and videos showing people who did not follow these recommendations (experiment 2) reduced unrealistic optimism. The third experiment, which included both articles and videos, replicated these results. These results can be applied to strategies for written and video communications that can be used by governments and public health agencies as best practices concerning not only COVID-19 but also any subsequent public health threat while promoting proactive, optimal, and healthy functioning of the individual.

Preparation of Sterile Raw Material - Chicken Eggshells in the Process of their Transformation into Selected Calcium Salts.

BACKGROUND: The chicken eggshells and their subcrustal membranes are a valuable source of calcium, but they are not further processed but disposed of as waste from the food industry. Chicken eggshells have high content (>95%) of calcium carbonate. Some properties suggest that eggshells may be a promising alternative to the present calcium sources used in the pharmaceutical industry. METHODS: The effect of roasting chicken eggshells with a selected organic acid (citric or fumaric or lactic acid) on microbiological purity, including the presence of fungi and bacteria Salmonella spp., Staphylococcus aureus, Escherichia coli of obtained calcium salts, was investigated. In this study, chicken eggshells were subjected to chemical reactions with organic acids (citric, fumaric or lactic acid) at two different calcium-acid molar ratios (1:1 or 1:3) and the mixture was heat-treated for 1 or 3 hours at a temperature of 100°C or 120°C. RESULTS AND DISCUSSION: It was found that lactic acid was 100% effective against fungi, and the remaining citric and fumaric acids were -50% (regardless of the other examined conditions). The type of acid used has a significant effect on fungal growth inhibition (p<0.05). Fumaric acid and lactic acid will be nearly 100% effective against bacteria (100% fumaric acid and 97% lactic acid effectiveness), regardless of other factors. CONCLUSION: Lactic acid is the most effective against pathogenic flora - fungi and bacteria. The transformation of chicken eggshells into calcium lactate can provide us with sterile calcium salt, free of 100% fungi and 97% of all bacteria.

Pre-Formulation Studies: Physicochemical Characteristics and In Vitro Release Kinetics of Insulin from Selected Hydrogels.

Insulin loaded to the polymer network of hydrogels may affect the speed and the quality of wound healing in diabetic patients. The aim of our research was to develop a formulation of insulin that could be applied to the skin. We chose hydrogels commonly used for pharmaceutical compounding, which can provide a form of therapy available to every patient. We prepared different gel formulations using Carbopol® UltrezTM 10, Carbopol® UltrezTM 30, methyl cellulose, and glycerin ointment. The hormone concentration was 1 mg/g of the hydrogel. We assessed the influence of model hydrogels on the pharmaceutical availability of insulin in vitro, and we examined the rheological and the texture parameters of the prepared formulations. Based on spectroscopic methods, we evaluated the influence of model hydrogels on secondary and tertiary structures of insulin. The analysis of rheograms showed that hydrogels are typical of shear-thinning non-Newtonian thixotropic fluids. Insulin release from the formulations occurs in a prolonged manner, providing a longer duration of action of the hormone. The stability of insulin in hydrogels was confirmed. The presence of model hydrogel carriers affects the secondary and the tertiary structures of insulin. The obtained results indicate that hydrogels are promising carriers in the treatment of diabetic foot ulcers. The most effective treatment can be achieved with a methyl cellulose-based insulin preparation.

Penetration of model hormones through the pericardium in simulated conditions in vivo.

The process of penetration of selected protein-peptide substances including insulin (INS), corticotropin (ACTH), prolactin (PRL) and albumin (reference protein) through the model membrane - pig pericardium was traced. These substances show a wide spectrum of therapeutic effects and diverse physicochemical properties (molecular weight, pI). The model substances penetrated the pericardium in simulated in vivo conditions from 1.0 mg / ml solutions. Based on the results obtained, pharmacokinetic parameters of the permeation process were determined - permeation rate (k), half-life (t50%) and their pharmaceutical availability (AUC [0-24 h]). All tested model substances penetrate the pericardium to different degrees. Within 24 h, they penetrate from 16.8% of albumin to 98.9% of insulin. Corticotropin penetrates 43.8% and PRL 34.2%. The highest availability is achieved with insulin, followed by ACTH, PRL and the lowest content of albumin. The results obtained suggest that the higher molecular weight of model protein-peptide substances, the lower the pericardial penetration (R2 = - 0.700) and availability (R2 = - 0.600), and the longer the half-life (R2 = 0.948).

Effect of Lutropin Concentration on the Efficiency of Isolated Porcine Kidney Storage in Modified Biolasol Solution.

BACKGROUND: Lutropin, a luteinizing hormone (LH), is a glycoprotein hormone secreted by gonadotropic cells of the anterior pituitary gland. LH receptors are a promising therapeutic target. The aim of the study was to analyze the protective effect of LH as a component of preservation solutions on the prevention of ischemia-reperfusion injury in isolated porcine kidneys. METHODS: The study used Biolasol solution (Biochefa, Sosnowiec, Poland) modified with the addition of ascorbic acid and LH at a concentration of 0.01 µg/L, 0.1 µg/L, and 1 µg/L. The study was carried out on 40 kidneys from 20 Polish large white pigs. Kidneys were divided into 4 groups, then washed out of the blood, and cooled by complete immersion in a 500-mL preservation solution at 4°C. The total storage time for grafts in the preservation solutions was 2 hours. After this time, the kidneys were perfused. Perfundates intended for biochemical analyses were collected from the renal vein after 0 minutes and after 30 minutes of perfusion. Another perfusion was performed after 24-hour renal storage. The activity of aspartate aminotransferase and alanine aminotransferase, as well as sodium, potassium, total protein, and urea concentrations was determined. RESULTS: LH as a component of preservation solutions affects the degree of ischemia-reperfusion injury of isolated porcine kidneys. CONCLUSIONS: The results obtained suggest that the efficacy of LH in improving ischemic kidney function is dependent on the dose used. The best graft protection was obtained using the lowest concentration of LH: 0.01 µg/L.

The Effectiveness of Rinsing Organs With Selected Preservation Solutions Based on the Analysis of Their Physicochemical Parameters.

BACKGROUND: One of the key elements of successful transplantation is effectively rinsing off the blood and preserving organs under controlled hypothermia. The aim of the study was to analyze the physicochemical parameters of Biolasol (Biochefa, Sosnowiec, Poland) and histidine-tryptophan-ketoglutarate (HTK) (Custodiol) solutions, which are recommended for perfusion and preservation of abdominal parenchymal organs. METHODS: Biolasol and HTK solution were used for the study. The solutions were subjected to physicochemical analysis involving pH, density, osmolarity, viscosity, refractive index, zeta potential, hydrodynamic diameter, and rheological properties. Rheological parameters were associated with morphologic features of fluids. RESULTS: HTK and Biolasol are non-Newtonian systems with pseudoplastic properties and yield stress. The solutions begin to flow under shear stress greater than the ultimate stress. In addition, a nonlinear relationship of their viscosity as a function of velocity gradient (shear rate) was observed. CONCLUSIONS: The solutions reproduce blood properties, which leads to the conclusion about their effective filling of the vascular bed and high efficiency in organ rinsing.