Histopathological Aspects of the Influence of Babesia microti on the Placentas of Infected Female Rats.

Babesiosis is perceived mainly an animal disease; however, awareness that Babesia spp. parasites that can cause diseases in humans is increasing significantly. Babesiosis is spread by the bite of an infected tick (Ixodes spp.), but it can also be transmitted by transfusion of infected blood and from an infected mother to her child during pregnancy or childbirth. The parasites multiply in the bloodstream and destroy red blood cells. This study aimed to assess the influence of Babesia microti on the histological structure of the placenta. Histopathological material collected from pregnant rats infected with Babesia microti was used in the experiment. Microscopic images of the placentas were assessed by Mallory staining and by using methylene blue-stained semi-thin sections. In addition, FISH was used to detect parasite DNA. The presence of piroplasms in both maternal and fetal vessels was demonstrated. Babesia microti infection caused vacuolization of syncytioblasts and cytotrophoblasts, accumulation of collagen fibers in placental villi, and increased adhesion of erythrocytes to the vascular walls. These results indicate that Babesia may influence the course of pregnancy and invite further research on the mechanism of piroplasm penetration into cells.

The Effect of Natural Substances Contained in Bee Products on Prostate Cancer in In Vitro Studies.

Prostate cancer is a common cancer in men in older age groups. The WHO forecasts an increase in the incidence of prostate cancer in the coming years. Patients may not respond to treatment, and may not tolerate the side effects of chemotherapy. Compounds of natural origin have long been used in the prevention and treatment of cancer. Flavonoids obtained from natural products, e.g., propolis, are compounds with proven antibacterial and antiviral efficacy which modulate the immune response and may be useful as adjuvants in chemotherapy. The main aim of the present study was to evaluate the cytotoxic and pro-apoptotic properties of selected flavonoids on prostate cancer cells of the LNCaP line. The compounds used in this study were CAPE, curcumin (CUR), and quercetin (QUE). Mitochondrial and lysosome metabolism was assessed by the XTT-NR-SRB triple assay as well as by the fluorescent staining techniques. Staining for reactive oxygen species was performed as well. The experiment showed that each of the tested compounds has a cytotoxic effect on the LNCaP cell line. Different types of cell death were induced by the tested compounds. Apoptosis was induced by quercetin, while autophagy-specific changes were observed after using CAPE. Compounds obtained from other bee products have antiproliferative and cytotoxic activity against LNCaP prostate cancer cells.

NiTi Shape Memory Clamps with Modified Surface for Bone Fracture Treatment.

Using NiTi alloys with shape memory for long-term medical implants requires modification of their surface due to the possible occurrence of corrosion. Hence, the surface of the staples used to join fractured bone within the craniofacial region was modified by applying a titanium oxy-nitrogen layer and a hydroxyapatite coating. Surface-modified clamps were tested in vivo using New Zealand white rabbits. After determining the mechanical characteristics of the bone and considering the initial state and surface modification, the diameter of the wire (used to make the clamps with the appropriate compression force) was selected. Implantation was performed on two groups of rabbits: experimental and control. In the experimental group, an intentionally induced bone fracture was treated in one tibia. On the second tibia, two additional clamps were applied to increase the possibility of a negative impact of the NiTi alloy on a living organism. After 6 weeks of application, a proper joining of the broken bone fragments was stated. Whereas after twelve weeks, no negative impact of the clamp material on a living organism, i.e., a rabbit, was found. Hence, the clamp with the modified surface can connect bone fragments in humans as well as small and medium-sized animals, with an extended range of use up to 12 weeks.

Histopathological Analysis of Selected Organs of Rats with Congenital Babesiosis Caused by Babesia microti.

Babesiosis is a tick-borne disease with an increasing number of cases each year. Due to the non-specific symptoms of babesiosis, insightful analyses of the pathogenesis of babesiosis are still very important. Transmission of the disease occurs in a few ways, which makes laboratory diagnosis of piroplasmosis important. Complications associated with the infection can be tragic, especially in patients with immunological disorders. The aim of this study was the histopathological analysis of the spleen and kidney of young Wistar rats infected transplacentally with Babesia microti. Female rats were infected with a reference strain of B. microti (ATCC 30221), and then, birth 3-week-old males were euthanized with isoflurane. Subsequently, the material was collected at autopsy for microscopic and ultrastructural examination. Microscopic and ultrastructural analysis of the spleen and kidney showed degenerative changes within the organ parenchyma and the capsules surrounding the organ. Regenerative and reparative changes through mitotic divisions of parenchymal cells were also evident. Merozoites of B. microti were visible in the section of erythrocytes and the cells building the organ stroma. The results presented in this study proved the negative effects of B. microti on cells and tissues in rats with congenital babesiosis.

Interactions between Babesia microti merozoites and rat kidney cells in a short-term in vitro culture and animal model.

Babesiosis is one of the most common infections in free-living animals and is rapidly becoming significant among human zoonoses. Cases of acute renal failure in humans caused by Babesia spp. have been described in the literature. The kidneys are characterised by intense blood flow through the blood vessels, which increases the likelihood of contact with the intra-erythrocyte parasite. The aim of this study was to observe the influence of B. microti (ATCC 30221) on renal epithelial cells in vitro cultured (NRK-52E line) and Wistar rats' kidney. Both NRK-52E cells and rats' kidney sections were analysed by light microscopy, transmission electron microscopy (TEM) and fluorescence in situ hybridization (FISH). Necrotic changes in renal epithelial cells have been observed in vitro and in vivo. In many cross-sections through the rats' kidney, adhesion of blood cells to the vascular endothelium, accumulation of erythrocytes and emboli were demonstrated. In NRK-52E culture, elements with a distinctly doubled cell membrane resembling B. microti were found inside the cytoplasm and adjacent to the cell layer. The study indicates a chemotactic tendency for B. microti to adhere to the renal tubules' epithelium, a possibility of piroplasms entering the renal epithelial cells, their proliferation within the cytoplasm and emboli formation.

Functional Severe Acute Respiratory Syndrome Coronavirus 2 Virus-Like Particles From Insect Cells.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) remains a major epidemic threat since the beginning of 2020. Efforts to combat the virus and the associated coronavirus disease 2019 (COVID-19) disease are being undertaken worldwide. To facilitate the research on the virus itself, a number of surrogate systems have been developed. Here, we report the efficient production of SARS-CoV-2 virus-like particles (VLPs) in insect cells. Contrary to widely used pseudovirus particles, where only one coronaviral protein is displayed within a heterologous scaffold, developed VLPs are structurally similar to the native virus and allow for more throughput studies on the biology of the infection. On the other hand, being devoid of the viral genome, VLPs are unable to replicate and thus safe to work with. Importantly, this is the first report showing that SARS-CoV-2 VLPs can be efficiently produced in insect cells and purified using scalable affinity chromatography.

Novel Antibacterial Modification of Polycarbonate for Increment Prototyping in Medicine.

In the era of modern medicine, the number of invasive treatments increases. Artificial devices used in medicine are associated with an increased risk of secondary infections. Bacterial biofilm development observed on the implanted surface is challenging to treat, primarily due to low antibiotics penetration. In our study, the preparation of a new polycarbonate composite, filled with nanosilver, nanosilica and rhodamine B derivative, suitable for three-dimensional printing, is described. Polymer materials with antimicrobial properties are known. However, in most cases, protection is limited to the outer layers only. The newly developed materials are protected in their entire volume. Moreover, the antibacterial properties are retained after multiple high-temperature processing were performed, allowing them to be used in 3D printing. Bacterial population reduction was observed, which gives an assumption for those materials to be clinically tested in the production of various medical devices and for the reduction of morbidity and mortality caused by multidrug-resistant bacteria.

Analysis of molecular and clinical parameters of 4-year adalimumab therapy in psoriatic patients.

INTRODCUTION: Through interaction with receptors TNFR1 and TNFR2, TNF-α activates a signal path, which exacerbates an inflammatory process, constituting an inseparable element of psoriasis. AIM: To evaluate changes in the expression of TNF-α, TNFR1, TNFR2 during the 4-year-long adalimumab therapy in psoriatic patients, searching for the correlation between molecular and clinical markers. In addition, the role of miRNAs was analysed. MATERIAL AND METHODS: Whole blood and serum samples of psoriatic patients treated with adalimumab constituted material for the study. Changes in the expression of TNF-α and its receptors were evaluated with the use of the RTqPCR method and MALDI ToF mass spectroscopy, PASI, BSA, DAS28 indexes were used for the clinical analysis of the patients, while the role of miRNA molecules was determined basing on microrna.org database. RESULTS: Different TNF-α expression patterns were determined in patients with observed resistance to the medicine. We found that there is a correlation between the molecular markers of an inflammatory process and the clinical indexes. The bioinformatic analysis indicates the potential role of miRNAs in the regulation of expression of the analysed genes. Changes in the profile of TNF-α during adalimumab therapy are significantly determined by the individual variability and susceptibility to the biological medicine or its loss. CONCLUSIONS: TNF-α seems to be a useful marker to evaluate the efficacy of therapy and occurring resistance to the medicine. A complex mechanism for the regulation of the analysed gene expression was underlined, which involved the potential role of miRNAs.

Caffeic Acid Phenethyl Ester (CAPE) Induced Apoptosis in Serous Ovarian Cancer OV7 Cells by Deregulation of BCL2/BAX Genes.

Ovarian cancer has the worst prognosis among all gynecological cancers. Therefore, it seems reasonable to seek new drugs that may improve the effectiveness of treatment or mitigate the adverse effects of chemotherapy. Caffeic acid phenethyl ester (CAPE) has many beneficial biological properties. The aim of the study was to assess the anticancer properties of CAPE against serum ovarian carcinoma cells. The morphology of the cells was evaluated in H-E staining and in transmission electron microscopy. The cytotoxic and proapoptotic activity of CAPE was investigated by using the XTT-NR-SRB assay, qRT-PCR analysis of BAX/BCL2 expression, and by cytometric evaluation. CAPE causes constriction in OV7 cells, numerous granulomas were observed in the cytoplasm, the cell nuclei were pyknotic. Autophagosomal vacuoles could suggest the occurrence of aponecrosis. CAPE significantly decreased the lysosomal activity and the total synthesis of cellular proteins. CAPE exhibited, dose and time dependent, cytotoxic activity against OV7 serum ovarian cancer cells. In OV7 cells CAPE induced apoptosis via dysregulation of BAX/BCL2 balance, while activated proapoptotic BAX gene expression level was 10 times higher than BCL2.

Membrane Protein of Human Coronavirus NL63 Is Responsible for Interaction with the Adhesion Receptor.

Human coronavirus NL63 (HCoV-NL63) is a common respiratory virus that causes moderately severe infections. We have previously shown that the virus uses heparan sulfate proteoglycans (HSPGs) as the initial attachment factors, facilitating viral entry into the cell. In the present study, we show that the membrane protein (M) of HCoV-NL63 mediates this attachment. Using viruslike particles lacking the spike (S) protein, we demonstrate that binding to the cell is not S protein dependent. Furthermore, we mapped the M protein site responsible for the interaction with HSPG and confirmed its relevance using a viable virus. Importantly, in silico analysis of the region responsible for HSPG binding in different clinical isolates and the Amsterdam I strain did not exhibit any signs of cell culture adaptation.IMPORTANCE It is generally accepted that the coronaviral S protein is responsible for viral interaction with a cellular receptor. Here we show that the M protein is also an important player during early stages of HCoV-NL63 infection and that the concerted action of the two proteins (M and S) is a prerequisite for effective infection. We believe that this study broadens the understanding of HCoV-NL63 biology and may also alter the way in which we perceive the first steps of cell infection with the virus. The data presented here may also be important for future research into vaccine or drug development.

Novel coronavirus-like particles targeting cells lining the respiratory tract.

Virus like particles (VLPs) produced by the expression of viral structural proteins can serve as versatile nanovectors or potential vaccine candidates. In this study we describe for the first time the generation of HCoV-NL63 VLPs using baculovirus system. Major structural proteins of HCoV-NL63 have been expressed in tagged or native form, and their assembly to form VLPs was evaluated. Additionally, a novel procedure for chromatography purification of HCoV-NL63 VLPs was developed. Interestingly, we show that these nanoparticles may deliver cargo and selectively transduce cells expressing the ACE2 protein such as ciliated cells of the respiratory tract. Production of a specific delivery vector is a major challenge for research concerning targeting molecules. The obtained results show that HCoV-NL63 VLPs may be efficiently produced, purified, modified and serve as a delivery platform. This study constitutes an important basis for further development of a promising viral vector displaying narrow tissue tropism.

Protective Effect of Polyphenol-Rich Extract from Bee Pollen in a High-Fat Diet.

We have studied a preventive effect of polyphenol-rich bee pollen ethanol extract (EEP) against histological changes in the liver and cardiac blood vessels, abnormalities of lipid profile, and the levels of oxidized low density lipoproteins (ox-LDL), asymmetric dimethylarginine (ADMA), angiotensin-converting enzyme (ACE), and angiotensin II (ANG II) caused by a high-fat diet in C57BL6 mice. Supplementing the diet with EEP in the doses of 0.1 g/kg body mass (BM) and 1 g/kg BM resulted in a decrease of total cholesterol by 31% and 35%, respectively. It also decreased the level of low density lipoproteins by 67% and 90%, respectively. No differences in the levels of high density lipoprotein and triacylglycerols were observed. EEP reduced the level of ox-LDL by 33% and 47%, ADMA by 13% and 51%, ACE by 17% and 30%, as well as ANG II by 11% and 15% in a dose-dependent manner, which proves a protective effect of EEP in a high-fat diet. EEP reduces and/or prevents hepatic steatosis and degenerative changes caused by a high-fat diet in C57BL6 mice, which indicates its hepatoprotective effect. EEP used with standard feed does not disturb a normal concentration of the assayed parameters.

Anti-Atherogenic Activity of Polyphenol-Rich Extract from Bee Pollen.

The aim of this study was to determine the effect of polyphenol-rich ethanol extract of bee pollen (EEP) on atherosclerosis induced by a high-fat diet in ApoE-knockout mice. EEP was given with feed in two doses of 0.1 and 1 g/kg body mass (BM). The studies have been conducted in a period of 16 weeks. The following factors were estimated: total cholesterol (TC), oxidized low density lipoproteins (ox-LDL), asymmetric dimethylarginine (ADMA), angiotensin-converting enzyme (ACE) and angiotensin II (ANG II) in the 5th, 10th, 12th, 14th, and 16th week of the experiment. In the last, i.e., 16th week of the studies the development of coronary artery disease (CAD) was also estimated histopathologically. Supplementing diet with EEP resulted in decreasing TC level. EEP reduced oxidative stress by lowering the levels of ox-LDL, ADMA, ANG II and ACE. EEP protected coronary arteries by significantly limiting the development of atherosclerosis (the dose of 0.1 g/kg BM) or completely preventing its occurrence (the dose of 1 g/kg BM). The obtained results demonstrate that EEP may be useful as a potential anti-atherogenic agent.

Induction of Apoptosis in Normal Human Dermal Fibroblasts Infected with Borrelia burgdorferi Sensu Lato.

The spirochete Borrelia burgdorferi s.l. can enter into different eukaryotic cells. Intracellular localization of bacteria may cause many changes in different cell pathways like apoptosis-mediated caspase cascade. The present studies focused on gene expression associated with caspase cascade after normal human dermal fibroblasts (NHDF) infection with Borrelia garinii, Borrelia afzelii, and B. burgdorferi s.s. The use of oligonucleotide microarray technique enabled an expression level comparison of genes associated with caspase cascade in NHDF infected with spirochetes. The increased expression of genes associated with caspase cascade was observed in case of CASP5, CASP2, CARD10, CASP10, MALT1, and NLRP1. The decreased expression was observed in case of CASP4, CASP6, and CASP1. The mRNA expression for CASP3 was inhibited in cells infected with three genospecies of Borrelia. However, the intensity of fluorescence was not statistically significant. In addition, cell cultures were fixed and procedure of caspase-3 detection and the TUNEL assay were performed. The in situ caspase-3 detection procedure confirmed the results obtained from microarray analyses. Only several fluorescent signals were observed. Many apoptotic cells were detected in NHDF-infected cultures with all spirochete genospecies found using the TUNEL reaction.

The epidemiological situation of chickenpox in the Silesian voivodeship in the years 2011-2015 in view of data of the situation in Poland / Sytuacja epidemiologiczna ospy wietrznej na terenie województwa slaskiego w latach 2011-2015 w swietle danych dotyczacych sytuacji w Polsce

INTRODUCTION: Despite the availability of preventive vaccination against chickenpox, this form of prevention is rarely used and is not available to the entire population of children. In order to evaluate an acquired immunity against the virus Varicella-Zoster examining of the presence of specific IgG antibodies to VZV in serum or plasma is required. The aim of this study was to assess the epidemiological situation of chickenpox in Slaskie Voivodeship in 2011-2015. MATERIAL AND METHODS: The evaluation of the epidemiological situation of chickenpox in the past five years was based on analysis of data from the bulletin "Infectious diseases and poisonings in Poland" for the period 2011-2014 and 2015 data, received from the Provincial Sanitary and Epidemiological Stations. Analysis of the vaccinated population was made on the basis of data available in the bulletin "Vaccinations in Poland" for the period 2011-2014 and 2015 data obtained from the NIPH-PZH. Samples of patients from Slaskie were tested with the use of Novalisa Varicella-Zoster Virus (VZV) IgG - ELISA (Novatec Immunodiagnostic GMBH, Germany). Samples were delivered to the Laboratory of the Provincial Sanitary and Epidemiological Station in the course of 2011-2015. RESULTS: Between 2011 and 2015, in Slaskie, 136 094 chickenpox cases were registered (14% of all occurring in Poland). Based on the number of cases, Slaskie is ranked second place, just after Mazowieckie, in which during the same period of time 143 392 illness were registered. The average annual incidence in Slaskie was 591 per 100 thousand residents. Between 2011 and 2015, a total of 360 serum samples were examined. The percentage of positive IgG in each year ranged between 59.6 and 75.7%. CONCLUSIONS: Promoting vaccination and preventing the sick children to contact the healthy ones as well as the protection of adults susceptible to infection can improve the epidemiological situation regarding incidences of chickenpox. Thanks to vaccines the risk of incidence of chickenpox can be reduced or even the incidences can be prevented. Information about acquired immunity, acquired before the pregnancy, allows to take the action in order to protect the mother from getting chickenpox in form of a preventive vaccination.

Congenital Tick Borne Diseases: Is This An Alternative Route of Transmission of Tick-Borne Pathogens In Mammals?

Tick-borne diseases (TBDs) have become a popular topic in many medical journals. Besides the obvious participation of ticks in the transmission of pathogens that cause TBD, little is written about alternative methods of their spread. An important role is played in this process by mammals, which serve as reservoirs. Transplacental transfer also plays important role in the spread of some TBD etiological agents. Reservoir species take part in the spread of pathogens, a phenomenon that has extreme importance in synanthropic environments. Animals that accompany humans and animals migrating from wild lands to urban areas increase the probability of pathogen infections by ticks This article provides an overview of TBDs, such as tick-borne encephalitis virus (TBEV), and TBDs caused by spirochetes, α-proteobacteria, γ-proteobacteria, and Apicomplexa, with particular attention to reports about their potential to cross the maternal placenta. For each disease, the method of propagation, symptoms of acute and chronic phase, and complications of their course in adults, children, and animals are described in detail. Additional information about transplacental transfer of these pathogens, effects of congenital diseases caused by them, and the possible effects of maternal infection to the fetus are also discussed. The problem of vertical transmission of pathogens presents a new challenge for medicine. Transfer of pathogens through the placenta may lead not only to propagation of diseases in the population, but also constitute a direct threat to health and fetal development. For this reason, the problem of vertical transmission requires more attention and an estimation of the impact of placental transfer for each of listed pathogens.

Hepatic tissue changes in rats due to chronic invasion of Babesia microti.

The etiological agents of babesiosis are intraerythrocytic parasites of the genus Babesia, which are transmitted by ticks. The course of disease is characterized by variable severity. The risk of a complicated course of babesiosis occurs in premature infants, the elderly, splenectomized patients and other immunocompromised patients. Severe cases of this disease can lead to multiple organ dysfunction. The study focuses on the impact assessment of chronic Babesia microti invasion on the morphology and ultrastructure of rat liver. The analyzed material was comprised of liver samples collected from Wistar rats infected with a reference strain of B. microti (ATCC 30221). None of the livers collected from rats with babesiosis was enlarged. The histopathological analyses showed signs of intensive inflammatory processes, especially in the perivascular areas. The hepatic mononuclear phagocyte system was characterized by increased activity. The ultrastructral analyses confirmed disintegration of hepatocytes with vacuolization in the perivascular areas. In addition, the perisinusoidal space (space of Disse) had irregular structure. In some areas, the space of Disse was enlarged or compressed. The morphological and ultrastructural analyses of rat liver with chronic babesiosis caused by B. microti showed significant pathological changes in perivascular areas which may be the cause of hepatic dysfunction.

Application of a real-time PCR method for Salmonella spp., Escherichia coli, Staphylococcus aureus and Clostridium perfringens detection in water samples.

The diagnostic assessment of water sanitary state is based mainly on the cultivation of bacteria retained on membrane filters. However classical microbiology methods have a lot of disadvantages. More and more frequently, rapid detection and identification of pathogens present in water is based on molecular biology techniques. The aim of this study was to determine the effectiveness and usefulness of a real-time PCR method, when compared to the recommended bacteria culture method, in diagnostics of pathogens in water samples. The research concerned the detection and identification of main sanitary indicators of water such as: Salmonella spp., Escherichia coli, Staphylococcus aureus and Clostridium perfringens. The analyses were conducted in water samples contaminated with the reference material (the aforementioned bacteria) and real environmental samples, which were examined for the presence of nucleic acid of: Salmonella spp., E. coli, S. aureus and C. perfringens using a real-time PCR method.

Nonspecific Bacterial Flora Isolated from the Body Surface and Inside Ixodes ricinus Ticks.

Ixodes ricinus and other representatives of the order Ixodida are vectors of typical pathogens: Borrelia burgdorferi sensu lato, Anaplasma phagocytophilium, Babesia spp., a tick-borne encephalitis virus, and other microorganisms which are important from a medical and veterinary point of view. The presented study focuses on the verification of nonspecific bacterial flora of I. ricinus. We analyzed ticks collected in a forest region in Silesia, an industrial district in Poland. Methods of classical microbiology and biochemical assays (API 20 NE test, API Staph test and MICRONAUT System) were used for isolation and identification of microorganisms living on the body surface of I. ricinus and inside ticks. The results show the presence of various bacteria on the surface and inside ticks' bodies. During the study, we isolated Acinetobacter lwoffi, Pseudomonas fluorescens, Aeromonas hydrophila, Achromobacter denitrificans, Alcaligenes faecalis, Stenotrophomonas maltophilia, Pseudomonas oryzihabitans, Micrococcus spp., Kocuria varians, Staphylococcus lentus, Kocuria kristinae, Streptococcus pneumoniae, Rhizobium radiobacter, Staphylococcus xylosus. Majority of the isolated species are non-pathogenic environmental microorganisms, but some of the isolated bacterial strains could cause severe infections.

The expression patterns of heat shock genes and proteins and their role during vertebrate's development.

Highly evolutionary conserved heat shock proteins (HSPs) act as molecular chaperones in regulation of cellular homeostasis and promoting survival. Generally they are induced by a variety of stressors whose effect could be disastrous on the organism, but they are also widely constitutively expressed in the absence of stress. Varied HSP expressions seem to be very essential in the critical steps of embryonic and extra-embryonic structures formation and may correspond to cell movements, proliferation, morphogenesis and apoptosis, which occur during embryonic development. While our knowledge of detailed HSP expression patterns is in constant progress, their functions during embryonic development are not yet fully understood. In the paper, we review available data on HSP expression and discuss their role during vertebrate development.