Histological and morphometrical evaluation of the urethral wall after bioresorbable stent implantation in male New Zealand White Rabbits: A preliminary study.

The aim of the study was the histological and morphometrical evaluation of the urethral wall at three time points after bioresorbable stent implantation in male New Zealand White Rabbits. The research was performed on 26 male New Zealand White rabbits aged 3-4 months and weighing 2.1-3.0 kg. Two models of bioresorbable sodium alginate-based stents were developed and implanted into the urethral lumen for one (T1), three (T3), and six weeks (T6). Sections of 5 µm thickness were cut from the urethra at intervals of 2 mm. The sliced sections were stained with hematoxylin-eosin (H&E), Van Gieson's (VG), Von Kossa, and Movat-Russell modified pentachrome (MOVAT) staining methods. The study provided valuable information for future models of urethral stents. The first model of the stent failed to fit the requirements due to inadequate mechanical properties. It curled up on itself losing the ability to adhere to the animals' urethra and was bioresorbed three weeks after implantation. The more rigid no. 2 stent was effective in widening the urethral lumen but did not biodegrade during the experiment. A comprehensive assessment of the second model's properties of biosorption and biointegration requires an extended observation of at least 12 months for an in depth morphological analysis. Stent migration is not likely to be caused solely by the mechanical properties of the urethra or urinary flow but mainly by muscle contraction of the organ wall.

Impact of early posthatch feeding on the immune system and selected hematological, biochemical, and hormonal parameters in broiler chickens.

Under commercial conditions, chicks hatch within a 24 to 48 h window, a period known as the hatching window. Subsequently, they undergo various treatments before finally being transported to the broiler farm. These procedures may delay the chicks' access to food and water, sometimes receiving them as late as 72 h after hatching. Previous studies have indicated that fasting during this initial period is detrimental, leading to impaired body growth, compromised immune system response, and hindered muscle development. The objective of this study was to assess the impact of early posthatch feeding on immune system organs and selected hematological, biochemical, and hormonal parameters. The experiment utilized Ross 308 broiler eggs incubated under typical commercial hatchery conditions. The experimental group's eggs were hatched in HatchCare hatchers (HC) with immediate access to feed and water, while the control group's eggs were hatched under standard conditions (ST). Thirty chickens from each group were assessed on the 1st (D1), 7th (D7), 21st (D21), and 35th (D35) day after hatching. On D1, the HC group exhibited lower hemoglobin, hematocrit, and total serum protein values, suggesting that early access to water prevents initial dehydration in newborn chicks. Conversely, the ST group showed a stress reaction on D1 due to feed deprivation, leading to an almost 2-fold higher serum corticosterone concentration compared to the HC group. However, this increase did not result in a significant change in the heterophil/lymphocyte ratio. Furthermore, the HC group displayed an increase in triglyceride concentration and a decrease in HDL concentration on D1. On D7, the HC group exhibited an increased relative weight of the bursa and a higher CD4+ cell number in the cecal tonsil (CT), indicating a more rapid development of these organs resulting from early stimulation of the gastrointestinal tract. However, early feeding did not influence the numbers of Bu-1+, CD4+, and CD8+ cells or the germinal center (GC) areas in the spleen. In conclusion, early feeding contributes to the welfare of newborn chicks by reducing dehydration and stress levels and stimulating the development of gut-associated lymphoid tissue.

The Effects of Honeysuckle (Lonicera caerulea L.) Berry Iridoid-Anthocyanin Extract on the Intestinal and Muscle Histopathology in Mice during Experimental Trichinellosis.

The aim of the study was to determine the effect of iridoid-anthocyanin extract from honeysuckle (Lonicera caerulea L.) (LC) berries on histopathological changes in the intestines and muscles during experimental trichinellosis in mice. The LC extract was administered to uninfected mice (LC group) and Trichinella-spiralis-infected mice (T+LC) orally at a dose of 2 g/kg bw, six times at 24 h intervals, from day 3 prior to infection to day 3 post-infection (dpi). Jejunum samples were collected on 5, 7, 14, and 21 dpi, and their histological assessment involved the villus height to crypt depth ratio (VH/CD), goblet cell (GC) number, and morphological changes. In the T. spiralis-infected muscles, the extent of inflammatory infiltration on the 14th and 21st dpi was assessed. LC in the infected mice restored the VH/CD ratio to control values on 14 dpi. A beneficial effect of the LC extract on the villus height was also observed 14 dpi in the LC and T+LC groups. No differences in the extent of inflammatory infiltration in the muscles between the T+LC and T groups were observed. In conclusion, the iridoid-anthocyanin extract from honeysuckle berry contributed to alleviating the symptoms of the intestinal phase of T. spiralis infection.

The cannabinoid receptors system in horses: Tissue distribution and cellular identification in skin.

BACKGROUND: The endocannabinoid system (ECS) is composed of cannabinoid receptors type 1 (CBR1) and type 2 (CBR2), cannabinoid-based ligands (endogenous chemically synthesized phytocannabinoids), and endogenous enzymes controlling their concentrations. Cannabinoid receptors (CBRs) have been identified in invertebrates and in almost all vertebrate species in the central and peripheral nervous system as well as in immune cells, where they control neuroimmune homeostasis. In humans, rodents, dogs, and cats, CBRs expression has been confirmed in the skin, and their expression and tissue distribution become disordered in pathological conditions. Cannabinoid receptors may be a possible therapeutic target in skin diseases. OBJECTIVES: To characterize the distribution and cellular expression of CBRs in the skin of horses under normal conditions. ANIMALS: Fifteen healthy horses. METHODS: Using full-thickness skin punch biopsy samples, skin-derived primary epidermal keratinocytes and dermal-derived cells, we performed analysis of Cnr1 and Cnr2 genes using real-time PCR and CBR1 and CBR2 protein expression by confocal microscopy and Western blotting. RESULTS: Normal equine skin, including equine epidermal keratinocytes and dermal fibroblast-like cells, all exhibited constant gene and protein expression of CBRs. CONCLUSIONS AND CLINICAL IMPORTANCE: Our results represent a starting point for developing and translating new veterinary medicine-based pharmacotherapies using ECS as a possible target.

The Differences in Histoarchitecture of Hoof Lamellae between Obese and Lean Draft Horses.

Obesity is a common problem in horses. The associations between obesity and equine metabolic syndrome (EMS) and between EMS and laminitis are known. However, there is a lack of data on whether obesity itself can affect hoof lamellae. Forelimbs and blood from 12 draft horses (six obese and six lean) from a slaughterhouse were acquired. To exclude laminitis and EMS horses, insulin concentration was measured, and hooves were radiographed. Histological evaluation was performed. The shape of the primary and secondary epidermal lamellae (PEL and SEL) was evaluated, and the length of the keratinized and total primary epidermal lamellae was measured (KPEL and TEL). All horses showed pathological changes in lamellae. In the lean group, the changes were longer SELs, more proliferated and separated PDLs, and less standard PDLs. In the obese group, the changes were a lower number of club-shaped and standard SELs and significantly more tapered SELs. No difference in the shape of PELs and the length of KPELs was noticed. The research did not confirm the effects of obesity on lamellar failure. The measurements taken indicate that the lamellae are much longer compared to other research studies; this could indicate that the length of the PEL depends on the hoof size.

The Improvement of the Adaptation Process of Tocopherol and Acetylsalicylic Acid in Offspring of Mothers Exposed to TCDD.

Dioxins are chemical compounds that may cause an inflammatory reaction. During dioxin-induced inflammation, generated reactive oxygen species lead to morphological changes in various tissues and in biochemical parameters. The aim of this study was to demonstrate the changes in the livers of rats whose mothers were exposed to dioxins and the protective role of α-tocopherol and acetylsalicylic acid in liver inflammation. The study material consisted of Buffalo rats who were the offspring of females treated with dioxin, dioxin + α-tocopherol, or dioxin + acetylsalicylic acid. Livers and blood samples were taken from the rats' offspring, and then histopathological and biochemical analyses were performed. The histopathological analysis showed that the changes observed in the livers of neonates were the result of the dioxins derived from their mother. The biochemical analysis showed that the morphological changes in the liver affected its function, which manifested in a higher total protein concentration in the dioxin-treated group, and that the creatinine level in this group was significantly higher than that in the other groups. This effect was reduced by the protective role of α-tocopherol and acetylsalicylic acid. Based on these results, we came to the conclusion that dioxins significantly affect the structure of the liver, which negatively affects its function, mainly in the scope of the metabolism of plasma proteins and hepatic enzymes.

Modification of a Haematoxylin, Eosin, and Natural Saffron Staining Method for The Detection of Connective Tissue.

INTRODUCTION: The aim of our study was to optimise an existing staining procedure: haematoxylin-eosin saffron (HES). The method follows the classical haematoxylin and eosin protocol with the addition of a staining step using natural saffron to better identify the collagen fibres. MATERIAL AND METHODS: The saffron solution was obtained by dissolving ground saffron stigmas in absolute alcohol. In order to test the HES method for its staining ability on four main types of collagen (I, II, III, and IV), specific tissues (skin, tooth, cartilage, aorta, spleen, and penis) were chosen. RESULTS: The procedure showed a sharp differentiation between muscle, stained red or pink, and connective tissue, stained bright yellow or orange. CONCLUSION: HES allows the diagnosis of reticulin fibrosis undetected in HE and in previous saffron staining procedures. HES represents an advantageous alternative to HE staining giving highly reproducible results with high diagnostic value.

Histological and morphometric evaluation of the urethra and penis in male New Zealand White rabbits.

This stereudy aimed at performing a histological and morphometric evaluation of the urethra and penis of male rabbits. Seven male New Zealand White rabbits weighing 2.1-3 kg were used in the study. The whole urethra, from the urinary bladder to the external urethral orifice, was dissected from the rabbits, and the tissue was sliced into sections at an interval of 2 mm. The sections were stained with haematoxylin-eosin, Masson-Goldner trichrome stain, Van Gieson's stain and Movat-Russell modified pentachrome stain. A detailed evaluation of the morphology and morphometry was performed. The parameters assessed were the type and height of epithelium, thickness and composition of connective tissue, and thickness and structure of muscularis. The histological structure of the rabbit urethra was found to be similar to humans. However, although the rabbits were found to have the same type of penis as the humans, the internal structure of the corpora cavernosa, the relative thickness of the tunica albuginea and the rudimentary glands of the penis were found to differ in these animals. The results of the present study may be useful in the designing of implants, drug testing or surgical procedures on the physiological and pathological urethra.

Impact of Prebiotics and Synbiotics Administered in ovo on the Immune Response against Experimental Antigens in Chicken Broilers.

The effect of the in ovo application of selected prebiotics and synbiotics on the humoral immune response against T-dependent (SRBC) and T-independent (dextran) antigens and delayed-type hypersensitivity (DTH) to phytohemagglutinin was studied. On the 12th day of incubation, 800 eggs (Ross 308) were divided into five groups and injected into the egg air chamber with prebiotic inulin (Pre1), Bi2tos (Pre2), a synbiotic composed of inulin and Lactococcus lactis subsp. lactis IBB SL1 (Syn1), a synbiotic composed of Bi2tos and L. lactis subsp. cremoris IBB SC1 (Syn2), and physiological saline (control group; C). The chickens were immunized twice at the 7th and 21st day of life with SRBC and dextran. A DTH test was performed on the 7th, 21st, and 35th day. The application of prebiotics and synbiotics had no significant effect on the humoral immune response. SRBC-immunized in ovo Pre1- and Pre2-treated chickens showed significantly higher serum IgG levels than the control. A significant effect on the DTH reaction was detected on the 7th (Pre1 < C) and 21st (Pre2 > Syn2) day. However; Bi2tos may transiently stimulate the cellular immune response on the 21st day. It may be concluded that the application of inulin in an egg air chamber on the 12th day of incubation may stimulate the secondary immune response. The inulin-treated group exhibited a lower mortality rate than the control group.

Therapeutic effects of an azaphenothiazine derivative in mouse experimental colitis.

Phenothiazines represent a class of compounds of potential therapeutic utility. In this report we evaluated therapeutic value of an azaphenothiazine derivative, 6-acetylaminobutyl-9-chloroquino[3,2-b]benzo[1,4]thiazine (QBT), given intragastrically, in the model of dextran sodium sulfate-induced colitis in C57BL/6 mice using 5-aminosalicylic acid (5-ASA) as a reference drug. Colitis symptoms such as body weight loss, diarrhea and hematochezia (blood in stool) were observed and registered and disease activity index (DAI) was calculated. In addition, weight and cell numbers in the lymphatic organs and histological parameters of the colon wall were analyzed. The effects of QBT on viability of colon epithelial cell lines were also determined. We showed that weight and cell number of draining mesenteric lymph nodes were lower in mice treated with QBT in comparison to their control counterparts. The number of thymocytes, drastically reduced in control mice, was elevated in mice treated with the compounds with a significant effect of 5-ASA. In addition, an abnormal composition of blood cell types was partially corrected in these groups. Histological analysis of the colon revealed that the pathological changes were partially normalized by QBT and even to a higher degree by 5-ASA. In conclusion we demonstrated a therapeutic efficacy of the compound in amelioration of local and systemic pathological changes associated with chemically-induced colitis in mice. A possible mechanism of action of the compound is discussed.

Accessory Genital Glands in the New Zealand White Rabbit: A Morphometrical and Histological Study.

INTRODUCTION: The aim of this research was to provide a detailed description of the morphology, topography, and histometry of rabbit accessory genital glands. MATERIAL AND METHODS: Seven male New Zealand White rabbits, 3-4 months of age and weighing 2.1-3 kg were used for the study. The whole urethra from the urinary bladder to the external urethral orifice accompanied by accessory genital glands was sliced at intervals of 1 mm. The serial sections were prepared with haematoxylin-eosin (H&E) and Movat-Russell modified pentachrome stain. RESULTS: A detailed description of the morphology and morphometry was provided. The topography of the organs was explained on the basis of characteristic cross-sections on histological slides. The inconsistent nomenclature and descriptions of these glands by different authors were also discussed. CONCLUSION: The morphometric analysis indicated that some of the glands described have similar dimensions in different individuals, while others like paraprostates revealed high diversity in the number of lobes, their size, and their structure. The accessory glands are also good topographic markers which precisely define the segment of the urethra. The terms "proprostate", "prostate", and "paraprostates" as the nomenclature of the prostate complex reflect the location of these glands well and indicate their common origin and function.

Selected prebiotics and synbiotics administered in ovo can modify innate immunity in chicken broilers.

BACKGROUND: A previous study showed that prebiotics and synbiotics administered in ovo into the egg air cell on the 12th day of incubation enhance the growth and development of chickens. However, the influence of this procedure on the development and efficiency of the innate immune system of broiler chickens is unclear. Therefore, the aim of this study was to evaluate whether the early (on the 12th day of embryo development) in ovo administration of selected prebiotics (inulin - Pre1 and Bi2tos - Pre2) and synbiotics (inulin + Lactococcus lactis subsp. lactis IBB SL1 - Syn1 and Bi2tos + L. lactis subsp. cremoris IBB SC1 - Syn2) influences the innate immune system. RESULTS: Chickens (broiler, Ross 308) that were treated with Pre1 exhibited a decreased H/L ratio on D7, but an increased H/L ratio was observed on D21 and D35. In the remaining experimental groups, an increase in the H/L ratio was observed on D21 and D35. The oxidative potential of leukocytes measured using the NBT test increased on D21 in Pre2 and Syn1 groups. The rate of the phagocytic ability of leukocytes increased in Pre1 and Syn1 groups on D21. The phagocytic index decreased in Pre1 and Syn2 groups on D21 and D35. Concurrently, the count of WBC in circulating blood decreased on D21 in Pre1, Pre2, and Syn1 groups. The hematocrit value was increased in Syn1 chickens on D21, in Pre1 chickens on D35, and in Syn2 chickens on both time points. CONCLUSIONS: Early in ovo treatment of chicken embryos with prebiotics and synbiotics may temporarily modulate not only the production/maturation of leukocytes but also their reactivity.

Topically applied azaphenothiazines inhibit experimental psoriasis in mice.

The therapeutic efficacy of topically applied azaphenothiazine derivatives: 9-chloro-6-acetylaminobutylquinobenzo[3,2-b][1,4]thiazine (compound 4) and 6-chloroethylureidoethyldiquino[3,2-b;2';3'-e][1,4]thiazine (compound 5) in the amelioration of inflammatory symptoms of imiquimod-induced psoriasis in mice was investigated. Clobederm®, containing clobetasol propioniate, served as a reference drug. The application of the compounds led to thinning of the epidermis and reduction of the cell layers. The suppressive actions of the compounds were even stronger with regard to pathological changes of the dermis. The compounds also exerted generalized, anti-inflammatory effects by decreasing the number of circulating leukocytes, lowering subiliac lymph node weight and partially normalizing an altered blood cell composition. The changes in the composition of main cell types in the epidermis and dermis were less affected by the compounds. In addition, both compounds inhibited to a similar degree production of tumor necrosis factor α (TNF α) in human whole blood cell culture. Whereas compound 5 strongly inhibited IL-8 and CXCL10 chemokines in human keratinocytes - KERTr cell line, transfected with poly(I:C), the suppressive action of compound 4 in this model was weak. In addition, compound 5, but not compound 4, exhibited at low doses proapoptotic properties with regard to colonic cell lines. In summary, we demonstrated the therapeutic potential of two selected azaphenotiazines in the amelioration of the skin pathology elicited in a mouse experimental model of psoriasis.

Topically applied azaphenothiazines inhibit contact sensitivity to oxazolone in mice.

In this work we investigated the efficacy of two topically applied azaphenothiazine derivatives, 9-chloro-6-acetylaminobutylquinobenzo[3,2-b][1,4]thiazine (compound 4) and 6-chloroethylureidoethyldiquino[3,2-b;2';3'-e][1,4]thiazine (compound 5), in the amelioration of inflammatory symptoms of contact sensitivity (CS) to oxazolone in mice, in relation to the commercial ointment Protopic® (tacrolimus), the reference drug. The compounds were administered 24 h following elicitation of CS and, 24 h later, the parameters of inflammation, such as ear edema, blood composition, leukocyte level, numbers of cells in the draining lymph nodes, histological picture of the inflamed tissue, and the morphometric analysis, were analyzed. The study showed that the effectiveness of the studied azaphenothiazines applied as a 0.1% ointment was comparable to the reference drug regarding suppression of the inflammatory process, when all the investigated histological parameters are taken into account.

Expression of hypoxia-inducible factor-1α and vascular density in mammary adenomas and adenocarcinomas in bitches.

BACKGROUND: The study aimed at examining hypoxia-inducible factor (HIF)1α expression in adenocarcinomas and adenomas in bitches in regard to tumour malignancy grade, proliferation, apoptosis and vascularisation. Therefore, paraffin sections of 15 adenomas and 64 adenocarcinomas sampled from 79 dogs aged 6 to 16 years were analysed. RESULTS: A significantly higher HIF-1α expression was noted in adenocarcinomas in comparison to adenomas (P < 0.0004). Moreover, HIF-1α expression in adenocarcinomas correlated positively with tumour malignancy grade (r = 0.59, P < 0.05), Ki-67 antigen expression (r = 0.43; P < 0.0005), TUNEL-positive cells (r = 0.62, P < 0001) and tumour vascularity measured by quantification of vessels characterized by the expression of von Willebrand Factor (r = 0.57, P < 0.05). CONCLUSION: Results of this study indicate a similar biological role of HIF-1α in dogs and in humans, which may confirm suitability of the animal model in investigations on progression of tumours in humans.

Serum and peritoneal evaluation of vitamin D-binding protein in women with endometriosis.

INTRODUCTION: Vitamin D-binding protein (also called DBP or Gc-globulin) is recognized as a multifunctional protein involved in the action scavenger system, the transport of vitamin D sterols, and the modulation of immune and inflammatory responses. This study evaluated total serum and peritoneal concentrations of vitamin D-binding protein in women with endometriosis, known as an inflammation-associated disease. MATERIALS/METHODS: The total concentration of DBP was measured with an enzyme-linked immunosorbent assay (ELISA) using a polyclonal antibody raised in a goat immunized with human DBP. Serum and peritoneal fluid were collected from women with endometriosis (n=26) and from patients with benign gynecological conditions serving as a control group (n=17). RESULTS: In general, the vitamin D-binding protein concentration was higher in serum than in peritoneal fluid. Women with endometriosis had higher serum but lower peritoneal levels of DBP compared with the control group; however, no significance was noted. When the endometriosis group was divided with regard to severity, an insignificantly higher serum level of DBP was observed in advanced endometriosis compared with the mild form of the disease, whereas the peritoneal concentration was not dependent on disease severity. CONCLUSIONS: It is concluded that serum and peritoneal DBP concentrations are not affected in women with endometriosis; however, based on the latest published data, it is possible that both the serum and peritoneal concentrations of vitamin D-binding protein may be dependent on Gc genotype, which results in differential modulation of monocyte/macrophage activity.