Cutaneous T‐cell lymphoma – Sézary syndrome in a Boxer

A 7‐year‐old male Boxer with a 3.5‐year history of atopy and food hypersensitivity was presented with multiple poorly circumscribed nodules and maculae of the skin and tongue, and jaundiced mucosal membranes. Cytologic and histopathologic examination of the skin lesions revealed cutaneous epitheliotropic lymphoma. Cells were CD3⁺ and CD8⁺ in flow cytometry. The CBC showed a moderate leukocytosis with 16% atypical lymphocytes with irregularly cleaved nuclei. Flow cytometric phenotyping of peripheral blood showed an elevated proportion of the CD8⁺ T‐lymphocyte subpopulation, indicating a malignant population of T‐cell origin, and the electropherogram of the PCR antigen receptor rearrangement produced a monoclonal peak for TCRγ. Liver enzyme activities were markedly increased and abdominal ultrasound examination showed increased echogenicity of the liver and enlarged abdominal lymph nodes. Fine‐needle aspirates of the liver confirmed infiltration with lymphocytes exhibiting the same morphology as the cells detected in skin and peripheral blood. Treatment was induced with L‐asparaginase, lomustine, and prednisone. Partial clinical remission of the skin and tongue lesions was achieved within 10 days, and hematologic abnormalities resolved. Despite further treatment with L‐asparaginase and lomustine, the dog relapsed within one month and was euthanized. Presence of malignant lymphocytes in skin, peripheral blood, and liver indicate a rare variant of leukemic cutaneous T‐cell lymphoma, equivalent of Sézary syndrome in a dog. This case report describes the use of flow cytometry as a complementary tool for lymphocyte characterization of skin lesions for the first time.     

Concentration profiles and safety of topically applied betulinic acid and NVX‐207 in eight healthy horses—A randomized,blinded, placebo‐controlled,crossover pilot study

The naturally occurring betulinic acid (BA) and its derivative NVX‐207 show anticancer effects against equine malignant melanoma (EMM) cells and a potent permeation in isolated equine skin in vitro. The aim of the study was to determine the in vivo concentration profiles of BA and NVX‐207 in equine skin and assess the compounds’ local and systemic tolerability with the intent of developing a topical therapy against EMM. Eight horses were treated percutaneously in a crossover design with 1% BA, 1% NVX‐207 or a placebo in a respective vehicle twice a day for seven consecutive days with a seven‐day washout period between each formulation. Horses were treated at the neck and underneath the tail. Concentration profiles of the compounds were assessed by high‐performance liquid chromatography in the cervical skin. Clinical and histopathological examinations and blood analyses were performed. Higher concentrations of NVX‐207 were found in the skin compared to BA. Good systemic tolerability and only mild local adverse effects were observed in all three groups. This study substantiates the topical application of BA and NVX‐207 in further clinical trials with horses suffering from EMM; however, penetration and permeation of the compounds may be altered in skin affected by tumors.     

Zum Bau der Milchdrüse des Rindes während der Trockenstehperiode unter besonderer Berücksichtigung der Involutionsprozesse

Light microscopical and histochemical investigations were performed on biopsies, which were obtained from the mammary glands of healthy pregnant cows during different stages of the nonlactating period. The results demonstrate characteristic morphological changes in the different phases of the nonlactating period. They can be interpreted as involution and regeneration processes, which occur in the alveoli and interalveolar tissue of the bovine mammary gland during the nonlactating period.     

Tissue Expansion in Pig Skin – A Histochemical Approach

In the present study, a porcine model for controlled skin expansion was investigated to improve our understanding of epidermal and vascular responses following stretching. The model is of outstanding importance not only for the clinical use of tissue expansion but provides interesting data for skin physiology and oncology, too. Thirteen out of 15 animals, who underwent silicone tissue expander implantation showed good clinical results. In all of them, skin biopsies were taken at the end of a controlled tissue expansion procedure (final expander volumes 350 or 500 ccm): one tissue specimen was obtained from the centre of the expanded skin area and a second from the neighbouring but nonexpanded skin. The tissue specimens were immediately frozen in liquid nitrogen and processed to 4 microns thick acetone-fixed frozen sections. Lectin histochemistry and immunohistology were performed using the following techniques: direct and indirect immunofluorescence technique (DIFT, IIFT), immunoperoxidase technique (POX) with either 3,3'-diamino-benzidine (DAB) or 3-amino-9-ethyl-carbazole (AEC). The histochemical findings were supplemented by measurements of the number of vital epidermal cell layers, the epidermal thickness (microns), and the papillary vascular count per visual field. There was a significant diminuation of the vascular count (mean +/- S.D. = 55.0% +/- 12.5%; U-test: p less than 5%). By immunohistochemistry, a loss of the basal cell reactivity for the following antibodies was noted: ACAM (against calmodulin), K 8.12 (against keratins 13 +/- 16) and A51-B/H4 (against keratins 8, 14, 18). There was a remarkable increase of filaggrin expression in the uppermost spinal cell layers in expanded skin, which was most pronounced in those specimens with the shortest interval to the last fluid injection into the expander. We gained no evidence for alterations of the expression of suprabasal epidermal keratins, lectin binding sites (UEA I, PNA, ConA, WGA), and vascular lectin- and immunoreactivity due to tissue expansion. The subdermal capsule, which had formed around the silicone expander, was strongly vimentin-reactive. In conclusion, controlled tissue expansion is capable to change the basal cell phenotype--a feature which is shared with a number of conditions with increased proliferative activity and with the epidermis covering different skin tumours. The regular expression of suprabasal keratins and epidermal lectin binding sites provides evidence for a normal epidermal cell differentiation. Furthermore, the porcine skin is a reliable model for studying physiology and pathophysiology of human skin.     

Einfluß der Ernährung auf den Papillarkörper der Pansenschleimhaut bei Kälbern

The purpose of this scanning electronmicroscopical study was to describe the structure of the lamina propria of the ruminal mucosa in calves on different diets.
The surface epithelium was removed with 0.3 % acetic acid. The exposed lamina propria showed structural differences, both macroscopically and microscopically in the different dietary groups. At the submicroscopic level, however, no differences were observed.     

Überprüfung der Anwendbarkeit eines Mikrotiter-Enzymimmuntests für Nachgemelk zur Fruchtbarkeitsanalyse bei der Kuh

Contents Evaluation of the reliability criteria of a Microtitre-Enzymeimmunoassay in strippings for fertility screening in the dairy cow The reliability criteria of a commercially available Milkprogesterone assay using microtitre plates were established and the applicability (including discriminatory levels) for fertility screening in strippings of the dairy cow was investigated. The sensitivity was found to be 2 pg/well and the following coefficients of variation were calculated: Interassay-variation (11–14%), intra-assay-variation (12–17%) and accuracy (12–17%). The assay is performed in less than three hours and provides reliable values to differentiate between samples from days 0 (AI), 6 and 21. The discriminatory levels are < 2 ng/ml milk (day 0), > 2 ng/ml (day 6) and > 10 ng/ml (day 21).     

Factors associated with canine skin extensibility in toy poodles

To assess factors for canine skin extensibility, our study investigated associations between the dogs’ skin extension index and the following factors, gender, age, neuter status, weight, coat color and six coat color related gene polymorphisms. Swab samples were collected from 69 toy poodles to extract DNA. The skin extension indices of the lower back and the neck were measured using the following formula: vertical height of the skin fold divided by body length multiplied by 100. The dogs’ age, weight, gender, neuter status and coat color were also recorded, as well as polymorphisms of the following six selected coat color related genes, Melanocortin 1 receptor, Tyrosinase-related protein 1, Melanophilin, Canine β-defensin-1, Major Facilitator Superfamily Domain Containing 12 and Agouti-signaling protein (ASIP). Univariable analysis showed there was a meaningful association between the lower back skin extension index and both gender and age (P<0.001 and P=0.048, respectively). Also, there was a possible association between the lower back skin extension index and ASIP Single nucleotide polymorphism (SNP) (R96C) (P=0.078). Linear model analysis showed there was a significant association between the lower back skin extension index and gender (P<0.001), and there was a tendency of the association between the lower back skin extension index and ASIP SNP (R96C) (P=0.098). In addition, there was an association between gender and age for the skin extension index. (P=0.048). Therefore, these results suggest that a greater risk of skin extensibility in toy poodle could be related to being female and the ASIP SNP (R96C), because these factors were associated with higher lower back skin extension index.     

Systemic Streptococcus agalactiae infection with meningo-ventriculitis in a Linnaeus’s two-toed sloth (Choloepus didactylus)

A captive male Linnaeus’s two-toed sloth died without any obvious clinical signs. At necropsy, multifocal ulceration at the lumbar and perianal skin, mitral valve vegetation, and multifocal hemorrhage in the leptomeninges were observed. Histopathologically, suppurative meningo-ventriculitis, dermatitis, and endocarditis characterized by severe neutrophilic infiltration were observed. Gram-positive cocci arranged in pairs or chains were present in these inflammatory lesions. Streptococcus agalactiae gene was detected in the skin, heart, and brain tissues by PCR and sequence analysis. These findings may indicate that S. agalactiae primarily infected the skin and then caused septicemia resulting in endocarditis and meningo-ventriculitis. The present case suggests that S. agalactiae infection can cause severe meningo-ventriculitis in two-toed sloth without any specific clinical signs.     

Oxidative stress in the pathogenesis of canine zinc‐responsive dermatosis

Zinc deficiency causes skin diseases both in humans and in animals. The underlying pathogenic mechanisms remain unclear, but a growing body of evidence indicates a role for zinc in skin protection against free radical‐induced oxidative damage. The immunohistochemical expression of heat shock proteins (HSPs; Hsp27, Hsp72, Hsp73 and Hsp90), Cu/Zn superoxide dismutase (SOD), metallothionein (MT), Ki‐67 antigen and active caspase‐3 were evaluated in normal canine skin and in samples from eight dogs with zinc‐responsive dermatosis. All investigated HSPs showed intense cytoplasmic immunostaining in the affected epidermis. Focal nuclear positivity of Hsp72 was also detected in keratinocytes. Although Cu/Zn SOD expression was similar to that observed in normal skin, MT immunoreactivity occurred in both the cytoplasm and the nucleus of basal cells in normal skin but was absent from the affected epidermis. Caspase‐3 activation was also absent in the involved epidermis, which revealed a high Ki‐67 index (a 3.5‐ to 9‐fold increase compared with normal skin). These results support the hypothesis that cellular response to stress, particularly oxidative stress, is involved in the pathogenesis of skin lesions in canine zinc‐responsive dermatosis. The lack of MT immunoreactivity in the affected epidermis may be indicative of low zinc levels, thus resulting in vulnerability to oxidative damage. In contrast, high expression levels of HSPs in skin during zinc deficiency may confer protection against a variety of dangerous stimuli, contributing to inhibition of apoptosis and to cell cycle regulation of proliferating keratinocytes.     

Das Blutgefäßsystem der Magenwand der Katze*

The blood vascular system of the stomach wall in the cat The results of 19 vascular-injections of the stomach wall in the cat are given. The arterial supply to the parietal surface is derived principally from the left gastric artery, that of the visceral surface from the short gastric branches of the splenic artery. Thus the arterial distribution is similar to the venous drainage. The blood supply of the tunica serosa and the tunica muscularis in the fornix and body regions arises from a common subserosal and muscular plexus. In the pyloric canal the blood supply arises from separate subserosal and intermuscular plexus. The submucosal plexus consists of both arteries and veins: from this, mucosal arteries ascend to a subglandular plexus and thence arterioles supply the capillary plexus of the lamina propria. Venules return the blood through collecting veins to a basal venous plexus, thence through mucosal veins to the submucosal plexus.     

Large granular lymphocyte lymphoma in the skin and urinary bladder of a dog

A 10-year-old female Cavalier King Charles Spaniel presented with hematuria, pollakiuria and skin rash. Based on the histopathological and cytological examination of the skin and bladder mucosa, the dog was diagnosed with large granular lymphocytic (LGL) lymphoma of the bladder and skin. The dog responded well to the initial chemotherapy with nimustine for 3 months. Since recurrence of skin erosion and bladder wall thickening were observed, the dog was subsequently administered chemotherapy with other anticancer drugs, including chlorambucil, vincristine, doxorubicin, L-asparaginase, cytosine arabinoside, and cyclophosphamide. The dog survived for 11 months and died due to tumor-related disseminated intravascular coagulation. This is the first report of a canine case of LGL lymphoma in the skin and bladder.     

Prevalence of Demodex canis‐positive healthy dogs at trichoscopic examination

Demodex canis is thought to be present in small numbers in the skin of most healthy dogs; however, available data on the prevalence of normal dogs harbouring D. canis are scarce. The purpose of this study was to investigate, using microscopic examination of plucked hairs, the prevalence of healthy dogs harbouring D. canis. Seventy‐eight clinically healthy dogs with no history of dermatological problems and clinically normal skin and hair coat were included in the study. Five areas (perioral skin 2–3mm from both labial commissures, periungual skin of the third digit of both anterior paws and chin) were examined in each dog. Fifty to sixty hairs were plucked from each skin site and microscopically examined. No D. canis mites were observed and only one adult form of Demodex injai was found in the labial commissure of one dog. Based on these results, the estimated prevalence of healthy dogs harbouring D. canis in clinically normal skin should not exceed the threshold of 5.4%, with 95% confidence level. Considering our and previous findings, we propose that, although small numbers of D. canis might inhabit the skin of normal dogs, the probability of finding these mites in normal dogs is low. Consequently, in most cases, the presence of a D. canis mite in the skin should not be considered as indicative of normality.     

Dermocystid infection in Japanese fire-bellied newt,Cynops pyrrhogaster

Here, we report details of a new infectious disease in wild-caught Japanese fire-bellied newts (Cynops pyrrhogaster), a Near Threatened species. Skin lesions consisting of numerous masses were found in the animals near Lake Biwa, Shiga Prefecture, Japan. The gross appearance of the skin lesions showed blister-, cyst-, and/or tumor-like morphology. Various sizes of skin lesions were observed on their entire body surface. Histologically, spherical basophilic cysts, including numerous spores, were observed in the dermis layer. Ultrastructural analysis indicated the presence of main bodies of flagellated zoospores within the spores. While 18s rRNA gene sequencing indicated that the skin lesions were due to dermocystid infection. To our knowledge, this is the first report of dermocystid infection in this amphibian in Japan. Further studies are needed to prevent epidemics and to establish diagnostic and treatment methods.     

Long‐Term Management with Adipose Tissue‐Derived Mesenchymal Stem Cells and Conventional Treatment in a Dog with Hepatocutaneous Syndrome

Hepatocutaneous syndrome (HS) is an uncommon skin disorder that occurs in conjunction with liver disease and is diagnosed based on decreased plasma concentrations of amino acids and the histopathology of skin lesions. The survival period generally is <6 months. A 10‐year‐old castrated male Maltese dog was presented for evaluation of lethargy, polyuria, polydipsia, and skin lesions including alopecia, erythema, and crusts. Based on increased liver enzyme activity, low plasma amino acid concentrations, and findings from liver cytology and skin biopsy, the dog was diagnosed with HS. In addition to administration of antioxidants, hepatoprotective agents, and amino acids IV, allogenic adipose tissue‐derived mesenchymal stem cells were infused 46 times over a 30‐month period: 8 times directly into the liver parenchyma guided by ultrasonography and the remainder of the times into peripheral veins. After commencing stem cell therapy, the dog's hair re‐grew and the skin lesions disappeared or became smaller. During ongoing management, the patient suddenly presented with anorexia and uncontrolled vomiting, and severe azotemia was observed. The dog died despite intensive care. On necropsy, severe liver fibrosis and superficial necrolytic dermatitis were observed. The dog survived for 32 months after diagnosis. A combination of amino acid and stem cell therapy may be beneficial for patients with HS.     

Cutaneous infiltrates and peripheral blood immune responses in dogs with immunomodulatory‐responsive lymphocytic–plasmacytic pododermatitis

This study characterizes T‐ and B‐lymphocyte responses in the peripheral blood and lesional skin of dogs with immunomodulatory‐responsive lymphocytic–plasmacytic pododermatitis (ImR‐LPP), a term previously proposed to denote a subpopulation of dogs with idiopathic pododermatitis. T‐cell (CD3⁺, CD4⁺ and CD8⁺) and B‐cell (CD21⁺) counts were significantly increased in both the epidermis and dermis of lesional ImR‐LPP skin compared with that in pedal skin from healthy controls. CD3⁺, CD4⁺, CD8⁺ and CD21⁺ cells were commonly observed in perivascular sites in the superficial dermis, periadnexally, beneath the dermal–epidermal (DE) junction and in the epidermis of lesional ImR‐LPP skin. The CD8⁺/CD3⁺ T‐cell ratio in peripheral blood was significantly increased in the ImR‐LPP group (0.42 versus 0.35 in controls). Serum IgA, IgG and IgM concentrations were all significantly elevated in affected dogs. Lymphocyte stimulation indices in ImR‐LPP dogs were comparable with control levels except for a lower response to ionomycin (6.0 versus 11.1). Dogs with ImR‐LPP had a higher incidence and mean (semi‐quantitative) score for IgA, IgG and IgM deposits in the epidermis, and a significantly increased incidence of dermal IgA⁺, IgG⁺ and IgM⁺ mononuclear inflammatory cells. The results indicate that upregulated T‐ and B‐lymphocyte responses may contribute to the pathogenesis of the skin lesions observed in dogs with ImR‐LPP.     

Vehicle Effects on the <Emphasis Type="Italic">In Vitro</Emphasis> Penetration of Testosterone through Equine Skin

The effects of three vehicles, phosphate-buffered saline (PBS), ethanol (50% in PBS w/w) and propylene glycol (50% in PBS w/w) on in vitro transdermal penetration of testosterone was investigated in the horse. Skin was harvested from the thorax of five Thoroughbred horses after euthanasia and stored at −20^°C until required. The skin was then defrosted and placed into Franz-type diffusion cells, which were maintained at approximately 32^°C by a water bath. Saturated solutions of testosterone, containing trace amounts of radiolabelled [¹⁴C]testosterone, in each vehicle were applied to the outer (stratum corneum) surface of each skin sample and aliquots of receptor fluid were collected at 0, 2, 4, 8, 16, 20, 22 and 24 h and analysed for testosterone by scintillation counting. The maximum flux (J _max) of testosterone was significantly higher for all sites when testosterone was dissolved in a vehicle containing 50% ethanol or 50% propylene glycol, compared to PBS. In contrast, higher residues of testosterone were found remaining within the skin when PBS was used as a vehicle. This study shows that variability in clinical response to testosterone could be expected with formulation design.     

Clinical and immunological heterogeneity of canine subepidermal blistering dermatoses with anti‐laminin‐332 (laminin‐5) auto‐antibodies

Laminin‐332 (laminin‐5) is a basement membrane heterotrimeric protein composed of alpha‐3, beta‐3 and gamma‐2 laminin chains. Laminin‐332 polypeptides are targeted by auto‐antibodies in human patients with mucous membrane (cicatricial) pemphigoid or, more rarely, subepidermal vesicular diseases that resemble epidermolysis bullosa acquisita (EBA) or bullous pemphigoid (BP). The objectives of this report were to characterize the clinical, histopathological and immunological characteristics of nine dogs with auto‐antibodies targeting laminin‐332. Immunological investigations consisted of direct immunofluorescence (IF), indirect IF with intact and salt‐split canine gingival, and salt‐split normal or laminin‐332‐deficient human skin, immunoblotting with purified human laminin‐332 and immunoblotting with recombinant NC1 domain of human collagen VII. All dogs exhibited varying degrees of skin blistering and ulceration associated with microscopic subepidermal vesiculation with or without inflammatory cells. Indirect IF established that circulating IgG auto‐antibodies bound the dermal side of salt‐split canine lip and human skin. In five dogs, IgG variably recognized the basement membrane of laminin‐332‐deficient human skin (three dogs negative, two dogs positive). In all nine dogs, IgG auto‐antibodies detected purified human laminin‐332 by immunoblotting. In two dogs, additional targeting of collagen VII‐NC1 was present. These observations establish laminin‐332 as a novel basement membrane antigen in dogs with autoimmune blistering diseases with variable clinical phenotypes. The names ‘acquired junctional epidermolysis bullosa’, ‘anti‐laminin‐332 mucous membrane pemphigoid (MMP)’ and ‘mixed auto‐immune subepidermal blistering dermatosis’ are proposed for dogs with clinical signs reminiscent of EBA, MMP or BP respectively.     

Tissue Engineering in Wound Repair: The three “R”s—Repair,Replace, Regenerate

Horses are predisposed to traumatic wounds that can be labor intensive and expensive to manage. Skin has a considerable potential for efficient and functional repair however, while cutaneous repair is a regenerative process in the fetus, this capability declines in late gestation as inflammation and scarring alter the outcome of healing. The historical gold standard for replacement of lost skin is the autologous skin graft. However, the horse's lack of redundant donor skin limits the practicality of full‐thickness grafting to smaller wounds; moreover, graft failure is relatively common in equine patients as a result of infection, inflammation, fluid accumulation beneath the graft, and motion. Tissue engineering has emerged as an interdisciplinary field with the aim to regenerate new biological material for replacing diseased or damaged tissues or organs. In the case of skin, the ultimate goal is to rapidly create a construct that effects the complete regeneration of functional skin, including all its layers and appendages. Moreover, an operational vascular and nervous network, with scar‐free integration within the surrounding host tissue, is desirable. For this to be achieved, not only is an appropriate source of cells required, but also a scaffold designed from natural or synthetic polymers. The newly created tissue might finally meet the numerous needs and expectations of practitioners and surgeons managing a catastrophic wound in a horse     

Untersuchungen zur Form der Schilddrüse (Glandula thyreoidea) beim Geflügel mit besonderer Berücksichtigung der jahreszeitlichen Schwankungen

The thyroid glands of 31 chickens at the age of 17 to 24 months were investigated. Different methods of anatomical preparation, casts of vessels and scanning electron microscopy were used. The thyroid gland of birds is a paired organ. It is located on the ventral surface of the base of the neck within the thoracic inlet. The left thyroid gland is placed more cranially than the right one. Each thyroid gland is closely connected to the common carotid artery on the medial side, from which it is supplied and to the jugular vein on the lateral side. It is a reddish-brown organ and of lenticular profile. The gland measures on average 10 mm in length, 6 mm in width and 2 mm in thickness, and is covered by a thin connective tissue capsule which holds adipose cells. It seems that each thyroid follicle is surrounded by a net of capillaries. The investigation by scanning electron microscopy proved that the follicles are oval with a pyramidal top on each end. The cuboidal epithelium cells leave impressions in the colloid. Epithelium cells carry microvilli on the follicle side surface. Described seasonal changes of the thyroid gland in size and activity were able to be confirmed by the examination of the organ in July and December. In winter the follicular cells were higher and the follicles had a greater volume.     

Zur Feinstruktur der normalen Kapillaren in der grauen Substanz der Großhirnrinde beim Schwein1

Fine structure of normal capillaries in the gray matter of the cerebral cortex of the pig The capillaries in the cerebral cortex over the Ammon's horn were studied with the electron microscope in 8 specific pathogen-free pigs. The capillaries had a thin continuous endothelium without perivascular space and their lumina on the average measured 5.8 ± 1.23 μm. in diameter. Their fine structure was by and large similar to that described for the capillaries of other species.