Among the main challenges in further advancing therapeutic strategies for Huntington’s disease (HD) is the development of biomarkers which must be applied to assess the efficiency of the treatment. HD is a dreadful neurodegenerative disorder which has its source of pathogenesis in the central nervous system (CNS) but is reflected by symptoms in the periphery. Visible symptoms include motor deficits and slight changes in peripheral tissues, which can be used as hallmarks for prognosis of the course of HD, e.g., the onset of the disease symptoms. Knowing how the pathology develops in the context of whole organisms is crucial for the development of therapy which would be the most beneficial for patients, as well as for proposing appropriate biomarkers to monitor disease progression and/or efficiency of treatment. We focus here on molecular peripheral biomarkers which could be used as a measurable outcome of potential therapy. We present and discuss a list of wet biomarkers which have been proposed in recent years to measure pre- and postsymptomatic HD. Interestingly, investigation of peripheral biomarkers in HD can unravel new aspects of the disease pathogenesis. This especially refers to inflammatory proteins or specific immune cells which attract scientific attention in neurodegenerative disorders.

Polyglutamine (polyQ) diseases are incurable neurological disorders caused by CAG repeat expansion in the open reading frames (ORFs) of specific genes. This type of mutation in the HTT gene is responsible for Huntington’s disease (HD). CAG repeat-targeting artificial miRNAs (art-miRNAs) were shown as attractive therapeutic approach for polyQ disorders as they caused allele-selective decrease in the level of mutant proteins. Here, using polyQ disease models, we aimed to demonstrate how miRNA-based gene expression regulation is dependent on target sequence features. We show that the silencing efficiency and selectivity of art-miRNAs is influenced by the localization of the CAG repeat tract within transcript and the specific sequence context. Furthermore, we aimed to reveal the events leading to downregulation of mutant polyQ proteins and found very rapid activation of translational repression and HTT transcript deadenylation. Slicer-activity of AGO2 was dispensable in this process, as determined in AGO2 knockout cells generated with CRISPR-Cas9 technology. We also showed highly allele-selective downregulation of huntingtin in human HD neural progenitors (NPs). Taken together, art-miRNA activity may serve as a model of the cooperative activity and targeting of ORF regions by endogenous miRNAs.

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Background Cystic endometrial hyperplasia-pyometra complex (CEH-P) is one of the most common uteropathies in bitches. In diseases with mild or obscure clinical signs and normal uterine size, a diagnosis based on a clinical assessment might be incorrect. The main aim of the research was to determine the morphological variables accompanying uterine diseases in bitches in microscopic evaluation. Consequently, the obtained results can be used to create a new classification system for uterine pathological changes during the development of the CEH-P, diagnosed by microscopic examination in bitches. Material for the study consisted of the uteri of 120 female dogs, aged 1–16 years, obtained during routine ovariohysterectomies. Macroscopic observation after a longitudinal incision of the uterine horns, allowed a preliminary classification of the uteri into research groups: control group (physiological uteri), and groups GI-III uteri collected form bitches with varying degrees of endometrial pathology. These preliminary classifications were then verified by histological analysis (H&E stain). Results The obtained results made it possible to determine and describe the prevalence (%) of pathological changes characteristic of the analyzed uterine diseases in the examined bitches. Histopathological analyses that were conducted have confirmed preliminary macroscopic evaluation for the control group, group GII (CEH), and group GIII (pyometra). In the uteri of the GI group, a severe congestion of the endometrium has been observed – this is typical of inflammation – which was not confirmed during histopathological examinations. However, these examinations revealed acute endometrial haemorrhage of varying severity. Conclusions Early reproduction disorders in bitches are, in general, not confirmed by clinical signs in the examined animals. The results show that during classification of typical morphological changes in the endometrium over the development of the CEH-P complex in bitches microscopic examinations are required. The obtained results indicate a frequent lack of consistency in the macroscopic assessment and histological analysis of the endometrium, observed in the analyzed uterine diseases, which in most cases is not followed by clinical symptoms. The presented classification of uterine diseases may be useful as a diagnostic tool in reproductive disorders in bitches and in examination in the field of basic research.

Cystic endometrial hyperplasia (CEH) and pyometra are the most frequently diagnosed uterine diseases affecting bitches of different ages. Transforming growth factor beta (TGF-β) has been classified in females as a potential regulator of many endometrial changes during the estrous cycle or may be involved in pathological disorders. The aim of this study was to determine the expression of TGF-β1, -β2 and -β3 in the endometrium of bitches suffering from CEH or a CEH–pyometra complex compared to clinically healthy females (control group; CG). A significantly increased level of TGF-β1 mRNA expression was observed in the endometrium with CEH–pyometra compared to CEH and CG. Protein production of TGF-β1 was identified only in the endometrium of bitches with CEH–pyometra. An increase in TGF-β3 mRNA expression was observed in all the studied groups compared to CG. The expression of TGF-β2 mRNA was significantly higher in CEH and lower in CEH–pyometra uteri. The results indicate the presence of TGF-β cytokines in canine endometrial tissues affected by proliferative and degenerative changes. However, among all TGF-β isoforms, TGF-β1 could potentially be a key factor involved in the regulation of the endometrium in bitches with CEH–pyometra complex.

Background The aim of the analysis was to designate the morphological symptoms that appear during the Cystic Endometrial Hyperplasia - Pyometra complex (CEH-P), diagnosed by microscopic examination. The investigation were conducted on the uteri of 120 bitches in age between 1–16. The microscopic examinations were based on histological stainings. The aim of examinations was to find the differences in morphology of endometrium in the specimens of the uteri wall (H&E stain). All of the uteri were divided into three pathological groups (GI – GIII), created on the basis of clinically symptoms of analyzed diseases. Uteri without symptoms were classified as a control group (C).Results Histopathological analysis that were conducted have confirmed preliminary macroscopic evaluation for control group with unchanged uteri, group GII with cystic endometrial hyperplasia of uteri (CEH), and group GIII with uteri with pyometra. The confirmation of compatibility of both macroscopic and microscopic evaluation of the uteri were observed in groups GII and GIII. In the uteri of the group GI a severe congestion of endometrium have been observed – it is typical for the inflammation – which was not confirmed during histopathological examinations. Those examinations revealed only endometrial haemorrhage.Conclusions The results are showing that during classification of research material microscopic examinations are required. The diagnose based on the macroscopic changes in typical for CEH-P symptoms might be incorrect, if it is not supported by detailed research. More than that, in all uteri with the closed pyometra the CEH was also observed. It suggest that pyometra may occur as a consequence of cystic endometrial hyperplasia and bacterial infections ended with inflammation. The results obtained can be used to create a basis for pathologic classification of endometrial hyperplasia, including CEH-P complex in bitches.

There are at least several ways of detecting bovine heat, which vary in accuracy, as well as in the effort and cost associated with the implementation and operation of equipment designed for this purpose. Heat in cows can be most easily detected by thorough and systematic observation of animals in the herd. Other methods include the measurements of vaginal mucosal resistance, the fitting of balloons with paint at the base of the tail or on the back, temperature measurements in the vagina, rumen and milk, and determination of progesterone levels in milk and blood. Currently, controlled breeding programmes, including timed artificial insemination (TAI) and automated estrus detection (AED) or activity monitoring system (AMS), are routinely introduced in high-performance dairy herds. Most of the commercially available heat detection devices are based on accelerometers or pedometers. These devices are capable of detecting heat in 81.4% to 91.3% of cows. The sensitivity of heat detection was 58.9% for accelerometers (Heatime), 63.3% for pedometers (SAE Afikim, Kibbutz Afikim, Israel), 56.7% for KaMaR markers and 35.9% for heat detectors (Scratchcard). When two of the three systems were combined (different combinations), the sensitivity increased to 75.9%. Similarly, after adding visual detection, sensitivity increased to 96% and specificity to 90% (for visual observation combined with AMS). Activity monitoring systems are a proven tool for improving the management of cows. They are an alternative to reproductive management based solely on traditional methods of detecting ovulation or hormone programmes (TAI) and they respect the well-being of animals. It seems, however, that further improvement in cows’ fertility on industrial cattle farms will be achieved by combining several different methods of heat detection and synchronization, rather than by using a single modern heat detection system..

Background: The cystic endometrial hyperplasia-pyometra complex (CEH-P) is one of the most common uteropathies in bitches. In diseases with mild or obscure clinical signs and normal uterine size, a diagnosis based on a clinical assessment might be incorrect. The main aim of the research was to determine the morphological variables accompanying uterine diseases in bitches in microscopic evaluation. Consequently, the obtained results can be used to create a new classification system for uterine pathological changes during the development of the CEH-P, diagnosed by microscopic examination in bitches. Material for the study consisted of the uteri of 120 female dogs, aged 1-16 years, obtained during routine ovariohysterectomies. Macroscopic observation after a longitudinal incision of the uterine horns, allowed a preliminary classification of the uteri into research groups: control group (physiological uteri), and groups GI-III uteri collected form bitches with varying degrees of endometrial pathology. These preliminary classifications were then verified by histological analysis (H&E stain).Results: The obtained results made it possible to determine and describe the prevalence (%) of pathological changes characteristic of the analyzed uterine diseases in the examined bitches. Histopathological analyses that were conducted have confirmed preliminary macroscopic evaluation for the control group, group GII (CEH), and group GIII (pyometra). In the uteri of the GI group, a severe congestion of endometrium has been observed – this is typical of inflammation – which was not confirmed during histopathological examinations. However, these examinations revealed acute endometrial haemorrhage of varying severity.Conclusions: Early reproduction disorders in bitches are, in general, not confirmed by clinical signs in the examined animals. The results show that during classification of typical morphological changes in endometrium over the development of the CEH-P complex in bitches microscopic examinations are required. The obtained results indicate a frequent lack of consistency in the macroscopic assessment and histological analysis of the endometrium, observed in the analyzed uterine diseases, which in most cases is not followed by clinical symptoms. The presented classification of uterine diseases may be useful as a diagnostic tool in reproductive disorders in bitches and in examination in the field of basic research.