Anatomical, Histological and Histochemical Observations of the Eyelids and Orbital Glands in the Lowland Tapir (Tapirus terrestris Linnaeus, 1785) (Perissodactyla: Ceratomorpha).

The lowland tapir is one of four species belonging to the Tapiridae family of the Ceratomorpha suborder, similar to Rhinocerotidae. This study describes anatomy with morphometry, histology (hematoxylin and eosin, Masson-Goldner trichrome, Movat pentachrome, mucicarmine, picro-Mallory trichrome) and histochemistry (PAS, AB pH 1.0, AB pH 2.5; AB pH2.5/PAS and HDI) of the upper and lower eyelids, and superficial gland of the third eyelid with the third eyelid, deep gland of the third eyelid, and lacrimal gland. The aim of the work is to show the features of the above-mentioned structures typical only for Tapiridae, as well as to show the presence of similarities and differences between the families forming the order Perissodactyla. The eyelashes on the upper eyelid were long, while those of the lower eyelid were short and much less prominent. In the upper and lower eyelid sebaceous glands, a characteristic simple alveolar gland producing a mucus-like secretion and poorly developed tarsal glands were observed. The marginal zone of the posterior surface of the eyelids was covered by stratified columnar epithelium with 18-21 layers of nucleated cells, while the bulbar zone of these surfaces was covered by cubic multilayer epithelium with 6-11 non-keratinized layers of cells and with sparse goblet cells. In only lower eyelids, numerous lymphoid nodules, diffuse lymphocytes and high endothelial venules were observed. The superficial gland was an acinar complex which secreted mucous and contained plasma cells within the interlobular and interlobular connective tissue. The upper and lower branches of the third eyelid were the shape of a bent "caudal fin" and were composed of hyaline cartilage, and they contained conjunctiva associated lymphoid tissue (CALT). The deep gland was also an acinar complex producing a serous character and having numerous diffuse lymphocytes. The lacrimal gland was an acinar complex producing seromucous secretions and had numerous plasma cells located in the glandular interstitium. The results of our research indicate that the features of the anatomy of the eyelids and orbital region in the lowland tapir are also typical of the family Tapiridae, but also have features common to the families Equidae and Rhinocerotidae. We confirm the presence of poorly developed tarsal glands in both eyelids as well as presence of a palpebral part of the lacrimal gland in the upper eyelid, which is typical only to Tapirus terrestris.

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.

Detection of Lymphatic Vessels in Dental Pulp.

The literature lacks conclusive evidence that lymphatic vessels can form in the dental pulp. Lymphangiogenesis is believed to occur in an inflamed pulp. If one defines lymphangiogenesis as the development of lymphatic vessels from already existing ones, such a mechanism is possible only when lymphatic vessels are present in healthy teeth. This paper aims to identify lymphatic vessels in the dental pulp using microscopic and immunohistochemical methods under physiological and pathological conditions. The tissue material consisted of human teeth intended for extraction. Our studies and results suggest a moderate correlation between pulp inflammation and the formation of new vessels, including lymphatic vessels.

Anti-Inflammatory Activity of a Cyclic Tetrapeptide in Mouse and Human Experimental Models.

A cyclic tetrapeptide Pro-Pro-Pheß3ho-Phe (4B8M) was tested for immunosuppressive activity and potential therapeutic utility in several in vitro and in vivo mouse and human models. The tetrapeptide was less toxic for mouse splenocytes in comparison to cyclosporine A (CsA) and a parent cyclolinopeptide (CLA). The tetrapeptide demonstrated potent anti-inflammatory properties in antigen-specific skin inflammatory reactions to oxazolone and toluene diisocyanate as well to nonspecific irritants such as salicylic acid. It also inhibited inflammatory processes in an air pouch induced by carrageenan. In addition, 4B8M proved effective in amelioration of animal models corresponding to human diseases, such as nonspecific colon inflammation induced by dextran sulfate and allergic pleurisy induced by ovalbumin (OVA) in sensitized mice. The tetrapeptide lowered expression of EP1 and EP3 but not EP2 and EP4 prostaglandin E2 (PGE2) receptors on lipopolysaccharide-stimulated Jurkat T cells and ICAM-1 expression on human peripheral blood mononuclear cells (PBMC). Its anti-inflammatory property in the carrageenan reaction was blocked by EP3 and EP4 antagonists. In addition, 4B8M induced an intracellular level of PGE2 in a human KERTr keratinocyte cell line. In conclusion, 4B8M is a low toxic and effective inhibitor of inflammatory disorders with potential therapeutic use, affecting the metabolism of prostanoid family molecules.

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.

Antidiabetic and Antioxidative Potential of the Blue Congo Variety of Purple Potato Extract in Streptozotocin-Induced Diabetic Rats.

This study was designed to evaluate the effects of purple potato extract of the Blue Congo variety (PP) on diabetes and its antioxidant activities after two-week administration tostreptozotocin (STZ)-induced diabetic rats. The activities of PP were evaluated at a dose of 165 mg/kg body weight (b.w.) by estimating biochemical changes in blood plasma and through a histopathological study of kidney, muscles, and liver tissue. We evaluated the effect of treatment with extract on glucose level, glycated hemoglobin, activities of enzymatic antioxidants (including superoxide dismutase, glutathione peroxidase, and catalase), and lipid peroxidation. Moreover, we determined advanced glycation end-products (AGEs), advanced oxidation protein products (AOPPs), and the level of oxidative modified proteins (OMPs) as markers of carbonyl-oxidative stress in rats with diabetes. Using high-performance liquid chromatography, we identified five anthocyanins and six phenolic acids in the extract from Blue Congo with the dominant acylated anthocyanin as petunidin-3-p-coumaroyl-rutinoside-5-glucoside. The administration of Blue Congo extract lowered blood glucose, improved glucose tolerance, and decreased the amount of glycated hemoglobin. Furthermore, PP demonstrated an antioxidative effect, suppressed malondialdehyde levels, and restored antioxidant enzyme activities in diabetic rats. After administration of PP, we also noticed inhibition of OMP, AGE, and AOPP formation in the rats' blood plasma.

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.

Homologous lactoferrin triggers mobilization of the myelocytic lineage of bone marrow in experimental mice.

The effects of lactoferrin (LF), an iron binding protein, on myelopoiesis have been studied extensively in vitro and in vivo in human and murine models over the past three decades. Due to the lack of high-quality homologous LFs, however, the conclusions are still unequivocal. Recently, both human and murine LFs have become available as recombinant products expressed in Chinese hamster ovary (CHO) cell lines showing mammalian type of glycosylation, thus apparently species compatible. In this study, we present the effects of homologous recombinant mouse LF (rmLF) on myelopoiesis in CBA mice. The myelocytic lineage has been assessed by their appearance in circulating blood and bone marrow, and induction of relevant mediators of inflammation. Intravenous injection of rmLF (100 µg/mouse) resulted in a significantly increased number of myelocytic cells in the circulating blood after 24 h. Mouse serum transferrin, used as a control protein, showed no stimulatory effect. The increase in output of neutrophil precursors, neutrophils, and eosinophils was correlated with a twofold increase of leukocyte concentrations. The analysis of the bone marrow sections confirmed increased myelopoiesis. The alterations in the bone marrow cell composition were statistically significant regarding mature neutrophils (10.8% vs. 27.7%), metamyelocytes (11.4% vs. 16.0%), and myelocytes (2.4% vs. 4.0%). The mobilization of the myelocytic cells in the bone marrow and the increased output of these cells into circulation were accompanied by elevated serum concentrations of interleukin-6 at 6 h and haptoglobin at 24 h following administration of rmLF. In conclusion, the homologous LF elicits significant and transient myelopoiesis in experimental mice.

Lactoferrin restrains allergen-induced pleurisy in mice.

OBJECTIVES: The aim of this study was to assess the utility of lactoferrin (LF), a natural immunomodulator, to restrain allergen-induced pleurisy in mice. MATERIAL AND SUBJECTS: BALB/c female mice, 8- to 10-week old, weighing 24 g on average, were used. TREATMENT: Mice were immunized intraperitoneally with 50 µg of ovalbumin (OVA) and the pleurisy was elicited 14 days later by intrapleural injection of 12.5 µg of OVA. LF was given 24 and 3 h before elicitation of the allergic reaction. METHODS: The cytokine levels in the pleural exudates were measured by immunoassays. The blood and pleural exudates smears were stained with Giemsa and May-Grünwald reagents and reviewed histologically. Lung sections were stained with eosin and hematoxylin for histological evaluation. RESULTS: Lactoferrin significantly decreased manifestation of pleurisy induced by OVA in a sensitized mouse model. In particular, the percentages of eosinophils in blood and pleural exudates were strongly diminished. The histological analysis of lungs revealed that LF diminished the development of pathological lesions, such as pulmonary edema, diffuse alveolar hemorrhage and hemosiderosis, which were found in the lungs after injection of the eliciting dose of OVA. LF also decreased the level of IL-5 secreted into the pleural fluid. CONCLUSIONS: This is a first demonstration that LF significantly decreases antigen-specific pleurisy in a sensitized mouse model.

17ß-Estradiol and interferon tau interact in the regulation of the immune response in a model of experimental autoimmune orchitis.

Immunoregulatory activity of type I interferons (IFNs) and estrogen is convergent in some cases of autoimmune disorders. The aim of our study was to determine whether a potent interaction of IFN and estradiol (E2) has an influence on immune response and estrogen receptor alpha (ER-?) expression in antigen-presenting cells in a model of experimental autoimmune orchitis (EAO). C3H/He/W male mice were immunized with testicular germ cells (TGCs) and orally treated with interferon tau (IFN-?), E2, or both simultaneously. The delayed-type hypersensitivity reaction was intensified after the administration of either IFN-? or E2, but their co-administration had no effect. IFN-? treatment increased immunoglobulin G2a (IgG2a) and decreased IgG1 levels of TGC-specific antibodies, whereas E2 abolished the effects of the used cytokine. The total splenic cellularity and the number of spleen CD11c+MHC II+ and F4/80+MHC II? cells were increased after IFN-? treatment, whereas E2 antagonized this effect. After IFN-? administration the level of ER-? was significantly higher in F4/80+MHC II? cells, whereas E2 had no effect. However, the administration of E2 significantly reduced the ER-? level in F4/80+MHC II+ and CD11c+MHC II+ cells in comparison with the IFN-??treated groups. In the EAO model, the type I IFN and E2 cooperated at the general and cellular levels of immune response, but E2 treatment usually abolished the effects exerted by the cytokine.

Determination of the mechanical properties of the skin of pig foetuses with respect to its structure.

Skin is an important barrier protecting the organism against external environmental factors. Determination of its mechanical characteristics as regards its structure has significant scientific and application value. In this work, uniaxial tensile tests were conducted to determine the basic mechanical parameters of skin with respect to its structure. The subject of the study were skin samples taken from domestic pig foetuses. They were excised from different parts of body, in the direction parallel to the long axis of the body. Regardless of the sampling site, the tests revealed no significant differences in the values of the maximum tensile strength (2.08 ± 0.25 MPa) and the conventional Young's modulus (5.87 ± 1.52 MPa). The mechanical and structural tests confirmed that regardless of the sampling region the skin of domestic pig foetuses may constitute a human skin substitute model.

Calf thymus extract attenuates severity of experimental encephalomyelitis in Lewis rats.

The aim of this study was to evaluate the efficacy of treatment of Lewis rats with calf thymus extract (TFX®) and its six-peptide fraction on the course of experimental allergic encephalomyelitis (EAE). Interferon- ß served as a reference drug. We found that intramuscular administration of the thymus extract fraction significantly reduced clinical, immunological, histological, and ultrastructural alterations inherent in the disease. We suggest that TFX® or TFX®-derived fractions have potential as therapeutics in treatment of neurodegenerative diseases such as multiple sclerosis.