4-Methylumbeliferone Treatment at a Dose of 1.2 g/kg/Day Is Safe for Long-Term Usage in Rats.

4-methylumbelliferone (4MU) has been suggested as a potential therapeutic agent for a wide range of neurological diseases. The current study aimed to evaluate the physiological changes and potential side effects after 10 weeks of 4MU treatment at a dose of 1.2 g/kg/day in healthy rats, and after 2 months of a wash-out period. Our findings revealed downregulation of hyaluronan (HA) and chondroitin sulphate proteoglycans throughout the body, significantly increased bile acids in blood samples in weeks 4 and 7 of the 4MU treatment, as well as increased blood sugars and proteins a few weeks after 4MU administration, and significantly increased interleukins IL10, IL12p70 and IFN gamma after 10 weeks of 4MU treatment. These effects, however, were reversed and no significant difference was observed between control treated and 4MU-treated animals after a 9-week wash-out period.

Initial screening of the rs104893657 variant of the PAX8 gene in women with hypothyroidism from Northeastern Slovakia.

OBJECTIVE: Thyroid diseases are among the most common endocrinopathies and metabolic disorders. Hypothyroidism is caused by insufficient production of thyroid hormones with a higher prevalence in women. Causes for the development of endocrine diseases may be mutations in genes that encode peptide hormones. The aim of this scientific study was to determine the genotype and allele frequencies of the rs104893657 variant of the PAX8 gene and to determine the genotype versus phenotype association. METHODS: The study population consisted of 135 women from northeastern Slovakia who were divided on the basis of screening into two groups: a control group without diagnosed hypothyroidism (CG = 67) and a group of women with hypothyroidism (HY = 68). Biochemical markers - thyroid-stimulating hormone (TSH), prealbumin (PREA), calcium (Ca), phosphorus (P), and alkaline phosphatase (ALP) were determined using Cobas Integra 400 plus, Cobas e411 analysers (Roche). Genotyping was performed using TaqMan® SNP Genotyping Assay instrument 7500 Fast Real-Time PCR Systems (Applied Biosystem). RESULTS: Student's t-test revealed a statistically significant difference between CG and HY in biochemical parameters: TSH (p < 0.001), P (p = 0.008). By Chi-square test we found no statistically significant difference in the representation of genotypes (p = 0.788) in the rs104893657 polymorphism of PAX8 gene. The T allele was not associated with hypothyroidism in Slovak women (p = 0.548). In CC genotype we found statistically significant difference between CG and HY in parameters TSH (p < 0.001) and P (p = 0.006). CONCLUSION: The mutant T allele was detected at low frequency in both groups of women studied. The association of the T allele with the development of hypothyroidism in Slovak women was not confirmed. The results of this work provide initial information on the distribution of genotypes and alleles in the studied variant of PAX8 gene in the Slovak female population.

Histopathology and indicators of borderline ovarian tumours with microinvasion in bitches.

The authors present two cases of borderline ovarian tumours with microinvasion in bitches with variable clinical significance. The first case documents a four-year-old female Weimaraner diagnosed with a tumour on the right ovary during a veterinary check-up, using ultrasound (USG) examination, which was then surgically removed. Histological examination revealed a clear cell borderline tumour of the ovary with microinvasion. The second case is represented by a necropsy sample from a 52-month-old female German Shepherd who died a day before the planned hysterectomy due to undertreated pyometra. During necropsy, a developed bilateral ovarian tumour was found. An additional histological examination revealed a serous borderline tumour with microinvasion of both ovaries. This paper discusses the histopathological and clinical aspects involved in the prognosis of borderline ovarian tumours in bitches. This concerns the possibility of a change for a more aggressive behaviour of these tumours and their immunohistochemical profile, then the risk of implant metastases and, finally, the time point of diagnosis, intervention, and therapy. Even histologically verified well-differentiated forms of borderline ovarian tumours with microinvasion in bitches can show the variable clinical significance and, therefore, in similar cases, only a good or only a bad prognosis of the disease should not be expected.

Suicides and their preventive interventions possibilities: are there some relationships between mechanisms and different mental disorders?

Suicides are one of the topics discussed around the world. This problem receives large space in scientific and professional literature, in order to eliminate its occurrence. Mechanisms of suicides are determined by whole spectrum of reasons determined by/depending on physical and psychological health. The objective of this work is to document the differences in mechanisms and realization of suicides by mentally sick people. Ten cases of suicides are reported in the article: three of which in people with a history of depression of the victim noted by family members, one with treated depression, three with anxiety-depressive disorder and three schizophrenic patients cases. There are five men and five women. Four of these women overdosed themselves by medicaments and one jumped out of a window. Two men shot themselves, two hanged themselves and one jumped out of a window. Persons without anamnestically proven psychiatric diseases end their lives mostly because of inconclusive situation or as a result of balancing their whole life (usually with good plan and preparation of the act). Persons with treatment of depression or anxiety-depressive disorder kill themselves mostly after several "unsuccessful" attempts. In case of victims with schizophrenia suicides follow a hardly predictable sequence of actions which sometimes do not seem to have any logic. Differences have been found between victims with and without mental disorders in ways of realization of suicides. Psychological predispositions in mood changes, long-term sadness and threatening suicide should be recognized by family members. Prevention of suicides in cases of people with a history of mental disorders is based on medical treatment and cooperation between the client and family members and a psychiatrist (Ref. 30). Keywords: forensic medicine, mental disorders, prevention, psychiatry, risk factors, suicides.

Oxidative Damage in Sporadic Colorectal Cancer: Molecular Mapping of Base Excision Repair Glycosylases MUTYH and hOGG1 in Colorectal Cancer Patients.

Oxidative stress, oxidative DNA damage and resulting mutations play a role in colorectal carcinogenesis. Impaired equilibrium between DNA damage formation, antioxidant status, and DNA repair capacity is responsible for the accumulation of genetic mutations and genomic instability. The lesion-specific DNA glycosylases, e.g., hOGG1 and MUTYH, initiate the repair of oxidative DNA damage. Hereditary syndromes (MUTYH-associated polyposis, NTHL1-associated tumor syndrome) with germline mutations causing a loss-of-function in base excision repair glycosylases, serve as straight forward evidence on the role of oxidative DNA damage and its repair. Altered or inhibited function of above glycosylases result in an accumulation of oxidative DNA damage and contribute to the adenoma-adenocarcinoma transition. Oxidative DNA damage, unless repaired, often gives rise G:C > T:A mutations in tumor suppressor genes and proto-oncogenes with subsequent occurrence of chromosomal copy-neutral loss of heterozygosity. For instance, G>T transversions in position c.34 of a KRAS gene serves as a pre-screening tool for MUTYH-associated polyposis diagnosis. Since sporadic colorectal cancer represents more complex and heterogenous disease, the situation is more complicated. In the present study we focused on the roles of base excision repair glycosylases (hOGG1, MUTYH) in colorectal cancer patients by investigating tumor and adjacent mucosa tissues. Although we found downregulation of both glycosylases and significantly lower expression of hOGG1 in tumor tissues, accompanied with G>T mutations in KRAS gene, oxidative DNA damage and its repair cannot solely explain the onset of sporadic colorectal cancer. In this respect, other factors (especially microenvironment) per se or in combination with oxidative DNA damage warrant further attention. Base excision repair characteristics determined in colorectal cancer tissues and their association with disease prognosis have been discussed as well.

Genetically modified rodent models and celiac, non-celiac gluten sensitivity: a minireview.

Celiac disease (CD) is a disorder that affects both children and adults. Over the few last decades, several new atypical cases have been identified through improved diagnostic tools. On the other hand, the onset of CD at a later age, including atypical CD forms whose clinical picture overlaps with other autoimmune diseases, shows that currently there are several unknown gene mutations, which could be responsible for the disease development. Non-celiac gluten sensitivity (NCGS) is entity included by the ingestion of gluten leading to intestinal, or extraintestinal symptoms that improve once the gluten is removed from the nutrition. In this article relationships between genetically modified rodent animals with previously unknown multiple organ changes and CD, respectively NCGS are reviewed. Relationships between the small bowel histological changes and other organs pathology are discussed. Results of research document that changes have similar genetic background and can develop to serious autoimmune systematic diseases, including small bowel inflammation resembling atypical CD or NCGS. These may have extra-intestinal symptomatology but without a clear explanation of causes and differences in their manifestations. Research on animal models helps to discover links between several disorders associated with gastrointestinal damage. New methods based on individual gene mutations can help in atypical adult CD and NCGS recognitions in the future.

Semi-Lethal Primary Ciliary Dyskinesia in Rats Lacking the Nme7 Gene.

NME7 (non-metastatic cells 7, nucleoside diphosphate kinase 7) is a member of a gene family with a profound effect on health/disease status. NME7 is an established member of the ciliome and contributes to the regulation of the microtubule-organizing center. We aimed to create a rat model to further investigate the phenotypic consequences of Nme7 gene deletion. The CRISPR/Cas9 nuclease system was used for the generation of Sprague Dawley Nme7 knock-out rats targeting the exon 4 of the Nme7 gene. We found the homozygous Nme7 gene deletion to be semi-lethal, as the majority of SDNme7-/- pups died prior to weaning. The most prominent phenotypes in surviving SDNme7-/- animals were hydrocephalus, situs inversus totalis, postnatal growth retardation, and sterility of both sexes. Thinning of the neocortex was histologically evident at 13.5 day of gestation, dilation of all ventricles was detected at birth, and an external sign of hydrocephalus, i.e., doming of the skull, was usually apparent at 2 weeks of age. Heterozygous SDNme7+/- rats developed normally; we did not detect any symptoms of primary ciliary dyskinesia. The transcriptomic profile of liver and lungs corroborated the histological findings, revealing defects in cell function and viability. In summary, the knock-out of the rat Nme7 gene resulted in a range of conditions consistent with the presentation of primary ciliary dyskinesia, supporting the previously implicated role of the centrosomally located Nme7 gene in ciliogenesis and control of ciliary transport.

Lethal alcohol intoxications cases: thinking about the causes.

Alcohol is a well-known, socially tolerated drug. Its harmfulness to human health, including its associated negative impact on the family of alcoholics, is well known. It is an interdisciplinary problem, whose solution requires financial contributions, society support, the involvement of a wider group of experts and finally also an active approach on young people and adults to this problem. The work is based on six lethal alcohol intoxication cases. The first case is devoted to a woman who dies in alcohol intoxication in the casino. The second case involves a man who died near the pond in alcohol intoxication. The third describes the course of the trip associated with drinking to the alcohol poisoning stage and the subsequent death of a man in a car. The fourth case is based on continual full-day drinking to the alcohol poisoning stage and subsequent death of a man. The fifth case describes the course of a riotous celebration, which ends with the death of an alcohol poisoned man. The last sixth case documents an alcohol poisoned man, which in combination with the ingestion of cannabinoids died on a bus station. The causes and reasons leading to such tragic events are discussed here. Attention is drawn to the dangers arising from the sudden drinking of occasional consumers and also to the dangers of the sudden drinking of persons who consume alcohol in excessive doses for a long time. Furthermore, the prevention of the use of alcoholic beverages by children and young people, including the prevention of alcoholism are discussed. Educational programs are proposed to create a remedial measure for de-tabooing this issue and format the attitudes of children, adolescents and adults to drinking alcohol, with the expectation of a reduction in deaths from alcohol intoxication in the future.

Oxidative Damage in Sporadic Colorectal Cancer: Molecular Mapping of Base Excision Repair Glycosylases in Colorectal Cancer Patients.

Oxidative stress with subsequent premutagenic oxidative DNA damage has been implicated in colorectal carcinogenesis. The repair of oxidative DNA damage is initiated by lesion-specific DNA glycosylases (hOGG1, NTH1, MUTYH). The direct evidence of the role of oxidative DNA damage and its repair is proven by hereditary syndromes (MUTYH-associated polyposis, NTHL1-associated tumor syndrome), where germline mutations cause loss-of-function in glycosylases of base excision repair, thus enabling the accumulation of oxidative DNA damage and leading to the adenoma-colorectal cancer transition. Unrepaired oxidative DNA damage often results in G:C>T:A mutations in tumor suppressor genes and proto-oncogenes and widespread occurrence of chromosomal copy-neutral loss of heterozygosity. However, the situation is more complicated in complex and heterogeneous disease, such as sporadic colorectal cancer. Here we summarized our current knowledge of the role of oxidative DNA damage and its repair on the onset, prognosis and treatment of sporadic colorectal cancer. Molecular and histological tumor heterogeneity was considered. Our study has also suggested an additional important source of oxidative DNA damage due to intestinal dysbiosis. The roles of base excision repair glycosylases (hOGG1, MUTYH) in tumor and adjacent mucosa tissues of colorectal cancer patients, particularly in the interplay with other factors (especially microenvironment), deserve further attention. Base excision repair characteristics determined in colorectal cancer tissues reflect, rather, a disease prognosis. Finally, we discuss the role of DNA repair in the treatment of colon cancer, since acquired or inherited defects in DNA repair pathways can be effectively used in therapy.

Anthropometric predictors of systolic and diastolic blood pressure considering intersexual differences in a group of selected schoolchildren.

OBJECTIVES: Although the association between anthropometric parameters and blood pressure has been established for adults, the relationship for children has not been thoroughly studied in Slovakia. Present study investigates the association between anthropometric parameters and systolic and diastolic blood pressure in the group of randomly selected schoolchildren. METHODS: Examinations were conducted as a cross-sectional study with 760 schoolchildren from Eastern Slovakia, 381 boys and 379 girls. The blood pressure evaluation included sphygmomanometer technique (seated, 3 times repeated) measurement of systolic and diastolic blood pressure (SBP; DBP). Anthropometric measurements included: body weight, height, circumference of waist, hip and chest, BMI, WHR (waist-hip ratio), fat skinfolds measurement, triceps skinfold, and derivation of body fat percentage. The parents' questionnaires incorporated basic demographic and socioeconomic characteristics of the family, reported BMI of father and mother, and child's birth-weight and birth-length. Statistical analysis included Student's t-test gender differences in measured parameters, partial Pearson's correlations and linear regression model of the impact of body parameters Z-scores on SBP and DPB. RESULTS: Statistical analysis confirmed gender difference in basic anthropometric parameters. Pearson's correlations revealed highly significant relationship of anthropometric indices to SBP compared to DBP. Correlations of anthropometric parameters with SBP and DPB were more significant for boys compared to girls. Linear regression analysis showed that the highest impact on SBP and DBP had Z-score of BMI, followed by weight and height. Z-score of body weight has statistical impact on SBP and DBP in all group and group of boys (p < 0.001), lower significance was in DBP of girls (p < 0.01). Similar results were obtained for Z-score of height and BMI for SBP and DBP in both total group and group of boys on level p < 0.001. Z-score for BMI and height in DBP has lower statistical significance. CONCLUSIONS: Linear regression model confirmed higher statistical relationship of SBP and DBP in the group of boys compared to the group of girls. SBP correlations and linear regression model of anthropometric parameters revealed more significant outputs compared to DPB. The results confirmed the fact that we have to consider anthropometric indices in paediatric blood pressure evaluation.

Compliance with gluten-free diet in a selected group of celiac children in the Slovak Republic.

OBJECTIVE: Celiac disease (CD) is a chronic long-term disease with autoimmune disorder connected to an allergic reaction to the molecule of gluten. CD is manifested in persons who are genetically predisposed. The only therapy for CD is a strong gluten-free (GF) diet. The aim of our study was to follow the adherence to the gluten-free diet in the selected group of parents and caregivers of children with CD, and to compare factors influencing the adherence to GF diet and therefore influencing also clinical symptoms of CD. We compared and examined important food-related activities as well, by using a personal questionnaire in the group of CD children's parents or caregivers. METHODS: We designed a cross-sectional study; 325 parents or caregivers of children with CD were recruited in the sample. CD was confirmed by the European Society for Paediatric Gastroenterology Hepatology and Nutrition (ESPGHAN) criteria. The anonymous questionnaires were collected at summer camps within period data 2012-2016 and followed factors which might have an influence on compliance with GF diet, according to parents' opinions. RESULTS: Adherence to GF diet was statistically significantly higher among girls compared to boys. Parents' questionnaires confirmed that younger group of children and children with the family history of CD had statistically higher compliance with GF diet. Parents with higher education confirmed better adherence to GF diet, statistical analyses confirmed higher adherence in mother´s education compared to father's education. Duration of CD, socio-demographic factors of parents - age of parent, gender of parent, and residency were not statistically significant in terms of compliance with the diet. CONCLUSIONS: Our study confirmed important factors influencing adherence to the GF diet, which are connected to the symptoms and consequences of CD. The assessment of parents' opinions and burden on the families of children with celiac disease should be also reflected in the evaluation of patient's status. Important is also the role of parents in order to facilitate and support clinical interventions and prevention of CD.

[Lethal cases of bleeding to the upper gastrointestinal tract]. / Letální prípady krvácení do horní cásti gastrointestinálního traktu.

We report three lethal cases of bleeding to the upper gastrointestinal tract as immediate cause of death. The first two cases are in connection with homelessness. First is a male who was found dead in a railway station, seated on a bench with dried blood beneath him. The second is a male who was found lifeless in a wooden shelter, where upon the good will of the landowners, he lived. At autopsy, gastroduodenal ulcer disease with a damaged blood vessel at the ulcer base was found in both. The cause of death was stated as hypovolemic shock, arising from loss of blood from bleeding gastroduodenal ulcer disease. The third case shows a man with liver cirrhosis, who was transported to hospital when his health deteriorated, where he died with blood vomiting. The case was closed as hypovolemic shock from loss of blood and was ascribed to haemorrhagic diathesis and chronic liver insufficiency, and excluded haemoptysis as the cause of the bleeding. The paper draws attention to the risk of mortality in patients with gastroduodenal ulcer disease, with irregular food intake, including alcohol abuse. The cases demonstrate the importance of medical care for patients who are prone to gastric ulcers, or to patients who have been diagnosed and/or treated for gastroduodenal ulcer disease, as the severe risk of ulcer bleeding, leading eventually to ulcer perforation. Our case reports may inform ambulance crews responding to at-risk individuals, such as homeless people, to the immediate threat of possible gastroduodenal ulcer disease with ulcer bleeding, or ulcer perforation.Key words: bleeding - death - gastric ulcer - gastroduodenal ulcer disease - nutrition - preventive medicine.

[Diabetic Kidney Disease 3rd stage - laboratory markers of mineral bone disorder]. / Diabetické ochorenie obliciek 3. stádia - laboratórne markery minerálovej a kostnej poruchy.

BACKGROUND: Diabetes mellitus is the most common cause of end stage kidney disease in the developed countries. Chronic kidney disease-mineral and bone disorder (CKD-MBD) develops with deteriorating of the renal functions. Diabetic patients on hemodialysis are characterized by low bone turnover, higher prevalence of severe and progressive vascular calcification with increased cardiovascular morbidity and mortality. The main factor which causes vascular calcification in patients with diabetic kidney disease (DKD) is poor glycemic control. The recent trial findings describe an inverse correlation between intact parathyroid hormone (iPTH) serum levels and glycemic control in a group of diabetic patients on hemodialysis. AIM: The objective of the proposed project is to access the difference of the laboratory markers MBD in the group of patients with 3rd stage DKD depending on glycemic control. We focused on the relationship between the glycemic compensation of diabetes (HbA1c) and iPTH serum level. PATIENTS AND METHOD: Ninety one patients with 3rd stage DKD were investigated. There were 46 women (50.5 %) and 45 men (49.5 %), average age of patients was 71.2 ± 7.0 years, with creatinine level 128 ± 30 µmol/l and estimated glomerular filtration (eGF, MDRD) 0.82 ± 0.16 ml/s. There were 60 patients with better glycemic control of diabetes (HbA1c < 7 %) vs 29 patients with poorly controlled diabetes (HbA1c > 7 %). MBD markers were compared in both groups. Patients were further stratified into subgroups based on the serum level of iPTH (iPTH < 35 pg/ml vs iPTH > 35 pg/ml) and MBD markers compared. Statistical analysis was performed using and Mann-Whitney test. RESULTS: We have found the statistical significance in the serum phosphate and proteinuria levels in between groups with HbA1c < 7 % vs patients with HbA1c > 7 %. Diabetics with better glycemic control had significant reduction in serum phosphate level (1.14 ± 0.20 vs 1.23 ± 0.18 mmol/l, p = 0.038) and in 24 hrs proteinuria level (0.56 ± 1.35 vs 1.30 ± 1.61 g/day, p = 0.007). In the group of presumed low bone turnover (iPTH < 35 pg/ml) we have found the trend towards increased serum calcium level (2.49 ± 0.12 vs 2.43 ± 0.10 mmol/l, p = 0.063) and increased HbA1c value (7.5 ± 1.8 vs 6.4 ± 1.6 %, p = 0.023). CONCLUSION: Our results suggest the closer relationship between glycemic control of diabetes and mineral-bone disorder in earlier stages of DKD. KEY WORDS: diabetes mellitus type 2 (DM2T) - chronic kidney disease (CKD) - mineral and bone disorder (MBD).

[Bloodstain pattern analysis on examples from practice: Are calculations with application parabolic trajectory usable?]. / Analýza krevních stríkancu na príkladech z praxe: Jsou výpocty po aplikaci parabolické trajektorie vyuzitelné?

The bloodstain pattern analysis (BPA) is useful in the forensic medicine. In Czechoslovakian criminology is this method not commonly used. The objective of this work is to calculate the impact length, height and distance splashing of blood drops. The results are compared with the real values for specific cases. It is also compared to calculate the angle of incidence of blood drops, using sinα with a form using tgα. For this purposes we used two different character cases from practice with well-preserved condition and readable blood stains. Selected blood stains were documented in order to calculate the angle of incidence of blood drops and to calculateorigin splashes. For this drop of blood, the distance of impact of the drops of blood (x), the height of the sprayed blood drops (y) and the length of the flight path the drop of blood (l). The obtained data was retrospectively analysed for the two models. The first straight line is represented by the triangle (M1) and the other is the parabolic model (M2). The formulae were derived using the Euler substitution. The results show that the angle of incidence of the drop of blood can be calculated as sinα and the tgα. When applying, the triangle is appropriate to consider the application and sinα parabolic requires the calculation of the angle of incidence drops of blood tgα. Parabola is useful for the BPA. In Czechoslovakian should be providing workplace training seminars BPA primarily intended for forensic investigators.We recommend the use of this method during investigations, verification of acts in forensic practice.

Effect of Green Tea on Weight Gain and Semen Quality of Rabbit Males.

The goal of the current study was to evaluate the action of the green tea plant (Camellia sinensis, L) on male rabbit reproduction and some non-reproductive indexes. Male rabbits were fed either a standard diet (control group) or a diet enriched with green tea powder (experimental groups; E): 5 g (E1) or 20 g (E2) per 100 kg of the milled complete feed mixture. Weight gain, sperm concentration, total and progressive motility, as well as haematological, and biochemical parameters and changes in testicular tissue histomorphology were evaluated. Feeding with green tea, at both tested concentrations, decreased weight gain per week and the total average weight gain compared to the control group (p < 0.05). Furthermore, green tea decreased sperm concentration, motility and progressive motility in the group fed with a lower dose (5 g) of green tea powder (p < 0.05), whilst a higher dose (20 g) was neutral. Some haematological and biochemical indexes, like medium-size cell count (MID), mean corpuscular haemoglobin concentration (MCHC), platelet percentage (PCT), levels of phosphorus (P) and total proteins (TP) were decreased in one or both experimental groups (p < 0.05), whilst the triglyceride level (TG) was increased in the E2 group (p < 0.05). The thicknesses of the testicular seminiferous tubules and epithelial layer were not affected by any concentration of green tea powder (p > 0.05). These observations suggest that green tea in the diet may have an adverse effect on rabbit growth and sperm quality, but their effect may be potentially dose-dependent.

Oral treatment of 4-methylumbelliferone reduced perineuronal nets and improved recognition memory in mice.

Hyaluronan (HA) is a core constituent of perineuronal nets (PNNs) that surround subpopulations of neurones. The PNNs control synaptic stabilization in both the developing and adult central nervous system, and disruption of PNNs has shown to reactivate neuroplasticity. We investigated the possibility of memory prolongation by attenuating PNN formation using 4-methylumbelliferone (4-MU), an inhibitor of HA synthesis. Adult C57BL/6 mice were fed with chow containing 5% (w/w) 4-MU for 6 months, at a dose ~6.7 mg/g/day. The oral administration of 4-MU reduced the glycosaminoglycan level in the brain to 72% and the spinal cord to 50% when compared to the controls. Spontaneous object recognition test (SOR) performed at 2, 3, 6 and 7 months showed a significant increase in SOR score in the 6-months treatment group 24 h after object presentation. The effect however did not persist in the washout group (1-month post treatment). Immunohistochemistry confirmed a reduction of PNNs, with shorter and less arborization of aggrecan staining around dendrites in hippocampus after 6 months of 4-MU treatment. Histopathological examination revealed mild atrophy in articular cartilage but it did not affect the motor performance as demonstrated in rotarod test. In conclusion, systemic oral administration of 4-MU for 6 months reduced PNN formation around neurons and enhanced memory retention in mice. However, the memory enhancement was not sustained despite the reduction of PNNs, possibly due to the lack of memory enhancement training during the washout period. Our results suggest that 4-MU treatment might offer a strategy for PNN modulation in memory enhancement.

Nematode-induced pathological lesions and alterations of mucin pattern identified in abomasa of wild ruminants.

Pathological lesions as well as mucin alterations in abomasa infected by nematodes have been thoroughly studied in livestock, but such data from wild ruminants are limited or completely lacking. Pathological data for Ashworthius sidemi, an invasive nematode are particularly rare. We necropsied the abomasa of 21 wild ruminants belonging to five cervid species and detected mixed nematode infections, dominated by A. sidemi. Samples from both gross lesions and mucous membranes without macroscopically apparent pathological alterations were subjected to standard histological procedures and histochemical staining. Histological examination found chronic abomasitis, manifested by edema, and hyperemia. Various degrees of lymphoplasmacytic infiltration were observed in all samples. Initial fibrosis (8/20, 40%) was detected in samples from both gross lesions and areas without macroscopically visible changes. Tissue from hemorrhagic lesions was superficially eroded. Generalized loss of surface polysaccharides was apparent in all samples. Only residual periodic acid-Schiff and Alcian blue (pH2.5) positivity was detected in the upper abomasal pits and in mucosal neck. This study found that nematode infections, mostly by A. sidemi, caused chronic inflammation and negatively affected abomasal mucin formation in wild ruminants.

The toxic influence of silver and titanium dioxide nanoparticles on cultured ovarian granulosa cells.

The application of metal nanoparticles in modern society is growing, but there is insufficient data concerning their influence on reproductive processes and comparison of their biological activity. The present experiments aimed to compare the effects of silver and titanium dioxide nanoparticles (AgNPs and TiO2NPs) on ovarian granulosa cell functions. AgNPs and TiO2NPs were added to culture of porcine granulosa cells at doses 0, 0.01, 0.1, 1 or 10 µg/mL. The mRNAs for proliferating cell nuclear antigen (PCNA), cyclin B1, bax and caspase 3 were quantified by RT-PCR; release of progesterone was analyzed by ELISA. It was shown that both AgNPs and TiO2NPs significantly reduced all the measured parameters. ED50 of the inhibitory influence of AgNPs on the main ovarian cell parameters was higher than ED50 of TiO2NPs. The ability of AgNPs and TiO2NPs to suppress ovarian granulosa cell functions should be taken into account by their application.

Immune activation by microbiome shapes the colon mucosa: Comparison between healthy rat mucosa under conventional and germ-free conditions.

Germ-free animals (GF) are those without a microbiome since birth. This particular biological model has become one of special interest with the growing evidence of importance of the microbiome in the life, development, adaptation, and immunity of humans and animals in the environments in which they live. Anatomical differences observed in GF compared with conventionally-reared animals (CV) has given rise to the question of the influence of commensal microflora on the development of structure and function (even immunological) of the bowel. Only recently, thanks to achievements in microscopy and associated methods, structural differences can be better evaluated and put in perspective with the immunological characteristics of GF vs. CV animals. This study, using a GF rat model, describes for the first time the possible influence that the presence of commensal microflora, continuously stimulating mucosal immunity, has on the collagen scaffold organization of the colon mucosa. Significant differences were found between CV and GF mucosa structure with higher complexity in the CV rats associated to a more activated immune environment. The immunological data suggest that, in response to the presence of a microbiome, an effective homeostatic regulation in developed by the CV rats in healthy conditions to avoid inflammation and maintain cytokine levels near the spontaneous production found in the GF animals. The results indicated that collagen scaffold adapted to the immune microenvironment; therefore, it is apparent that the microbiome was able to condition the structure of the colon mucosa.

Local Immune Changes in Early Stages of Inflammation and Carcinogenesis Correlate with the Collagen Scaffold Changes of the Colon Mucosa.

Continuous activation of the immune system inside a tissue can lead to remodelling of the tissue structure and creation of a specific microenvironment, such as during the tumour development. Chronic inflammation is a central player in stimulating changes that alter the tissue stroma and can lead to fibrotic evolution. In the colon mucosa, regulatory mechanisms, including TGF-ß1, avoid damaging inflammation in front of the continuous challenge by the intestinal microbiome. Inducing either DSS colitis or AOM colorectal carcinogenesis in AVN-Wistar rats, we evaluated at one month after the end of each treatment whether immunological changes and remodelling of the collagen scaffold were already in development. At this time point, we found in both models a general downregulation of pro-inflammatory cytokines and even of TGF-ß1, but not of IL-6. Moreover, we demonstrated by multi-photon microscopy the simultaneously presence of pro-fibrotic remodelling of the collagen scaffold, with measurable changes in comparison to the control mucosa. The scaffold was significantly modified depending on the type of induced stimulation. These results suggest that at one month after the end of the DSS or AOM inductions, a smouldering inflammation is present in both induced conditions, since the pro-inflammatory cytokines still exceed, in proportion, the local homeostatic regulation of which TGF-ß1 is a part (inflammatory threshold). Such an inflammation appears sufficient to sustain remodelling of the collagen scaffold that may be taken as a possible pathological marker for revealing pre-neoplastic inflammation.