Interacting He and Ar atoms: Revised theoretical interaction potential, dipole moment, and collision-induced absorption spectra.

Coupled cluster quantum chemical calculations of the potential energy surface and the induced dipole surface are reported for the He-Ar van der Waals collisional complex. Spectroscopic parameters are derived from global analytical fits while an accurate value for the long-range dipole coefficient D7 is obtained by perturbation methods. Collision-induced absorption spectra are computed quantum mechanically and compared with existing measurements.

Effect of dietary zinc oxide on morphological characteristics, mucin composition and gene expression in the colon of weaned piglets.

The trace element zinc is often used in the diet of weaned piglets, as high doses have resulted in positive effects on intestinal health. However, the majority of previous studies evaluated zinc supplementations for a short period only and focused on the small intestine. The hypothesis of the present study was that low, medium and high levels of dietary zinc (57, 164 and 2,425 mg Zn/kg from zinc oxide) would affect colonic morphology and innate host defense mechanisms across 4 weeks post-weaning. Histological examinations were conducted regarding the colonic morphology and neutral, acidic, sialylated and sulphated mucins. The mRNA expression levels of mucin (MUC) 1, 2, 13, 20, toll-like receptor (TLR) 2, 4, interleukin (IL)-1ß, 8, 10, interferon-γ (IFN-γ) and transforming growth factor-ß (TGF-ß) were also measured. The colonic crypt area increased in an age-depending manner, and the greatest area was found with medium concentration of dietary zinc. With the high concentration of dietary zinc, the number of goblet cells containing mixed neutral-acidic mucins and total mucins increased. Sialomucin containing goblet cells increased age-dependently. The expression of MUC2 increased with age and reached the highest level at 47 days of age. The expression levels of TLR2 and 4 decreased with age. The mRNA expression of TLR4 and the pro-inflammatory cytokine IL-8 were down-regulated with high dietary zinc treatment, while piglets fed with medium dietary zinc had the highest expression. It is concluded that dietary zinc level had a clear impact on colonic morphology, mucin profiles and immunological traits in piglets after weaning. Those changes might support local defense mechanisms and affect colonic physiology and contribute to the reported reduction of post-weaning diarrhea.

Keratinization of the esophageal epithelium of domesticated mammals.

We studied the esophageal epithelium for keratinization characteristics from samples of domesticated mammals of three nutrition groups (herbivores: horse, cattle, sheep; omnivores: pig, dog, rat; carnivores: cat) using histochemistry (keratins, disulfides), sulfur measurements, and cryo-SEM. Keratins were found in all esophageal layers of all species, except for the equine Stratum corneum. The positive reaction staining of Pan-keratin was remarkable, but decreased in intensity toward the outer layers, whereas in the pig and cat, staining was confined to the corneal layer. The herbivores revealed positive staining reactions in the upper Stratum spinosum, particularly in the sheep. Regarding single keratins, CK6 immunostating was found in most esophageal layers, but only weakly or negatively in the porcine and equine Stratum corneum. CK13 staining was restricted to the sheep and here was found in all layers. CK14 could be detected in the equine and feline Stratum basale, and upper vital layers of the dog and rat. CK17 appeared only in the Stratum spinosum and Stratum granulosum, but in all layers of the dog and cat. Disulfides reacted strongest in the Stratum corneum of the herbivores, as corroborated by the sulfur concentrations in the esophagus. Our study emphasized that keratins are very important for the mechanical stability of the epithelial cells and cell layers of the mammalian esophagus. The role of these keratins in the esophageal epithelia is of specific interest owing to the varying feed qualities and mechanical loads of different nutrition groups, which have to be countered.

Immunohistochemical demonstration of connexins in the developing feather follicle of the chicken.

Based on immunohistochemistry, the study demonstrates the varying distribution and reaction intensity of connexins (Cx26 [chicken 31sim], 30 [chicken 31], 31, 32, 43, 45) in the developing feather follicle of the chicken (White Leghorn). The different embryonal stages were identified according to the normal table of Hamburger and Hamilton (1951). The development of the feather follicle complex is closely related to skin layer development, making use of the controlling function of connexins. This was evident during feather follicle differentiation, based on communication between ectomesodermal (fibroblasts) and ectodermal cells (developing epidermis), but also by the subsequent separation of the two cell line types related to their connexin-dependent differentiation degree. With the increase in mesenchymal cell numbers during feather placode development, the multiple connexins Cx26 [chicken 31sim] and 43, supported by Cx30 [chicken 31], 31 and 32, were increasingly activating the fibroblast concentrations as related to epidermal follicle buds, the specific follicle structure, the endothelial cells of capillaries and larger blood vessels, as well as the collagen fiber production and the growing feather musculature shortly before hatching; Cx45 could not be demonstrated. In conclusion, it seems that connexin expression is not only coupled to the origin of embryonic cells, but also connected with tissue formation before the follicle system can be formed.

Method for the preparation of mucosal flaps from the jejunum of laying hens for transporter studies in Ussing chambers.

Ussing chambers are frequently used for in vitro evaluation of intestinal transport physiology. The current study describes investigating the jejunal tissue from laying hens using a specific preparation method and evaluates the effect of glutamine in the maintenance buffer. Tunica mucosa was stripped from 104 jejunal samples from 10 hens and stabilised by a net device. Fifty samples were maintained with modified Krebs-Henseleit buffer (Control), 54 samples with additional 5 mM glutamine (Group Gln). The percentage of responding samples varied between 87 and 100%. Mean short circuit current (ΔI sc,) [µA/cm(2)] of samples exposed to 10 mM glucose in the Control group and Group Gln was 17.0 and 14.6 (p = 0.836), respectively, of samples exposed to 100 µM phloridzin -13.3 and -11.8 (p = 0.712), respectively, and of samples exposed to 100 µM carbachol 4.7 and 3.7 (p = 0.450), respectively. In conclusion, the net-supported method enabled a reliable investigation of jejunum from laying hens. Glutamine in the maintenance buffer was of no significant benefit.

Cytochemistry of sialoglycoconjugates and lysozyme in canine anal glands as studied by electron microscopic methods.

The distribution of sialoglycoconjugates and lysozyme in the secretory cells of canine anal glands was studied by means of electron microscopic cytochemical methods, particularly lectin cytochemistry and immunocytochemistry. Sialic acids were predominantly present in the secretory granules, Golgi bodies, surface coat of the plasma membrane and luminal secretions. In addition, within these structures, the secretory granules, Golgi bodies and luminal secretions exhibited high levels of sialoglycoconjugates that terminated in Siaα2-6Gal/GalNAc or Siaα2-3Galß1-4GlcNAc. In the secretory cells, reactive gold particles representing lysozyme were mainly detectable in the secretory granules and Golgi bodies. Sialic acids possess diverging functional properties, whereas lysozyme contributes to the non-specific defense against microorganisms. Therefore, their presence and secretion are suggestive of protective effects of both secretory products at the anal mucosa.

Cytochemistry of sialoglycoconjugates, lysozyme, and ß-defensin in eccrine glands of porcine snout skin as studied by electron microscopy.

In most mammals except for humanoid primates, eccrine glands are confined to the skin of a series of specific body regions. Sialic acids and antimicrobial substances exhibit various functional properties and serve as a component of nonspecific defense against micro-organisms, respectively. In this study, the distribution of these moieties was studied by electron microscopic histochemical methods. The eccrine glandular acini consisted of two types of dark cells as well as clear cells. The secretory granules and Golgi apparatus of both types of dark cells contained sialic acid residues linked to α2-6Gal/GalNAc. On the other hand, sialoglycoconjugates with Siα2-3Galß1-4GlcNAc sequence were confined to those of the Type II dark cells. In addition, lysozyme and ß-defensin were mainly detected in the secretory granules of the Type II dark cells. These secretory products may create a defensive barrier against microbial invasion and play an essential role in preservation of the integrity of porcine snout skin as a sensory organ.

Sialic acids and antimicrobial substances in the apocrine glands of porcine perianal skin.

The porcine perianal skin shows prominent apocrine glands with large saccular dilatations, whereby the functional significance of the glandular secretions is rather unexplained. Our study focuses on the demonstration of sialoglycoconjugates and antimicrobial substances in these glands, using glycoconjugate histochemical and immunohistochemical methods. The result obtained emphasized the general presence of sialic acids, linked to α2-6Gal/GalNAc and α2-3Gaßl1-4GlcNAc, in the secretory cells. The secretory epithelium and luminal secretions also contained a spectrum of antimicrobial substances, such as lysozyme, IgA, lactoferrin, and the peptide group of ß-defensins. Realizing that sialic acids possess diverging functional properties through various saccharide residues, and that antimicrobial substances serve as a non-specific defense against microorganisms, these secretory products may function as protective agents in order to preserve the integrity of the perianal region. This view includes that the amounts of bacteria on the skin surface are controlled and maintained at the certain level.

A special construction of subepidermal capillary loops in the hippopotamus (Hippopotamus amphibius).

Based on LM, TEM, and histochemical methods, the study describes the specific structure of subepidemal capillary loops in the integument of the hippopotamus (Hippopotamus amphibius). At 25- 60 µm, the diameter of the capillaries was more than twenty times larger than those found in other mammals, as was the diameter of the epidermal contact area of the hairpin turn, which had enlarged up to 200-400 µm(2). At about 13,400, the number of loops per cm(2) was three times higher than in the few other mammalian species measured to date. The remarkable sheath (thickness 2- 20 µm) of the capillary loops consists of a multitude of fine collagen IV fibres, which were in direct contact with the epidermal stratum (str.) basale, emphasizing an origin from the lamina fibroreticularis of the basement membrane. Additionally, the sheath contained many regions filled with free fatty acids. All observations confirmed the view that the walls of the subepidermal capillaries in the hippopotamus are adapted to withstand high blood pressure, permitting a high rate of blood vesselbased heat transfer from the periphery of the body. Until now this function is only known as an important thermoregulatory response in highly active mammals, e.g. dolphins. However, under hot climatic conditions but without strong exercise for cooling, such ability could be an effective and energy-saving procedure in semi-aquatic mammals.

Histochemical distribution of sialic acids and antimicrobial substances in porcine carpal glands.

The localization of sialic acids and antimicrobial products (lysozyme, IgA, lactoferrin, ß-defensin 2) as well as Rab3D in the carpal glands of pig was studied by sialoglycoconjugate histochemistry and immunohistochemistry. The secretory epithelium of the carpal glands consisted of dark and clear cells. The dark cells of these glands exhibited high levels of sialoglycoconjugates, including O-acetylated sialic acids, whereas the localization of sialic acids linked to α2-3Gal1-4GlcNAc was confined to a subpopulation of the dark cells. Furthermore, all antimicrobial substances and Rab3D were mainly detectable in a subpopulation of the dark cells. The results obtained are discussed with regard to the functional significance of these glands. Our findings suggested that Rab3D is involved in the secretory regulation of sialoglycoconjugates and antimicrobial substances. These secretory products may create a defensive barrier against microbial invasion and play an essential role in the preservation of skin integrity.

Demonstration of free fatty acids in the integument of semi-aquatic and aquatic mammals.

The sensitive red fluorescence dye BODIPY® 665/676, and embedding in the water-soluble resin Technovit® 7100 were used to demonstrate free fatty acids in the epidermis of seven semi-aquatic and aquatic mammalian species with a sparse or dense hair coat. The staining generally marked lipid layers of varying thickness between the lamellae of the Stratum corneum, as found particularly in very densely haired species (otter), but also in rather sparsely haired animals (beaver, nutria), and especially in the seal. The very sparsely haired capybara contained no free fatty acids in the corneal layer system, but exhibited an accumulation of such substances in the vital epidermis. All haired species showed a strongly positive reaction staining of the sebaceous glands. In the hairless species, a distinct intracellular staining was restricted to cells of the thick vital epidermis in the hippopotamus, whereby in the Str. corneum positive intercellular reactions appeared. In the dolphin, on the contrary, positive intercellular reactions became visible in the vital epidermis, whereas in the Str. corneum the lipids concentrated in large longitudinal intracellular vesicles.

Histochemical analyses of glycoconjugates and antimicrobial substances in goat labial glands.

Saliva is known to protect the oral cavity and contains glycoproteins and antimicrobial substances. The distribution of these salivary secretions was studied in the labial glands of the Japanese miniature (Shiba) goat using lectin histochemical and immunohistochemical methods. The mucous acinar cells of the labial glands exhibited glycoconjugates with different saccharide residues, such as GalNAcα1-3GalNAc, Galß1-4GalNAc, ß-D-GlcNAc and sialic acid linked to α2-6Gal/GalNAc. Furthermore, α-D-Man, α-L-Fuc, α-D-GalNAc, ß-D-Gal and sialic acid residues were present, in particular, in the serous demilunar cells. Antimicrobial substances (lysozyme, IgA, lactoferrin and ß-defensin) were shown to be mainly immunolocalized in the serous demilunes and duct cells. The results obtained are discussed with regard to the functional role of labial glands. The secretory compounds demonstrated may play an important role in the maintenance of oral health with regard to saliva.

Histochemical demonstration of sialic acids and antimicrobial substances in the porcine anal glands.

The distribution of sialic acids and antimicrobial products (lysozyme, ß-defensin-1, lactoferrin, IgA) in the anal glands of miniature pig was studied by glycoconjugate histochemistry and immunohistochemistry. The glandular acini of these glands exhibited considerable amounts of sialoglycoconjugates that terminated in Siaα2-6Gal/GalNAc or Siaα2-3Gal1-4GlcNAc, including O-acetylated sialic acids. Additionally, all antimicrobial products examined could be demonstrated in the anal glands, especially in the serous cells. The results obtained are discussed with regard to the functional significance of the anal glands. Our observations corroborated the view that sialic acids closely interact with defense cells and antimicrobial substances in the innate immune response. Therefore, the anal glandular secretions may function as protective agents in order to preserve the integrity of the anal region.

Functional morphology of the tongue in the domestic goose (Anser anser f. domestica).

Using LM and SEM methods, the study describes microstructures in particular areas of the tongue of the goose. A thick multilayered keratinized epithelium forms the "lingual nail" and covers small and giant conical papillae, whereby the first functions as an exoskeleton of the tongue apex, and the latter are arranged along the lingual and well-developed connective tissue cores, and together with the bill lamellae are involved in cutting. The row of conical papillae on the lingual prominence prevents regurgitation of transported food. In the area of the "lingual nail" and in the anterior part of the lingual prominence, Herbst corpuscles are accumulated, which allow to recognize food position. Filiform papillae, as widely distributed between the conical papillae of the body, are responsible for filtering. They can be explained as long keratinized processes of the epithelium and are devoid of connective tissue cores. During food transport, the flattened areas of the lingual body and the lingual prominence are protected by a parakeratinized epithelium, but the root is covered by a nonkeratinized epithelium. The presence of adipose tissue in the tongue probably reduces pressure during food passage, but also promotes mucus evacuation from the lingual glands, thus facilitating food transport. An entoglossal bone with a continuation as cartilage is the stable structural basis of the tongue system.

Major translocation of calcium upon epidermal barrier insult: imaging and quantification via FLIM/Fourier vector analysis.

Calcium controls an array of key events in keratinocytes and epidermis: localized changes in Ca(2+) concentrations and their regulation are therefore especially important to assess when observing epidermal barrier homeostasis and repair, neonatal barrier establishment, in differentiation, signaling, cell adhesion, and in various pathological states. Yet, tissue- and cellular Ca(2+) concentrations in physiologic and diseased states are only partially known, and difficult to measure. Prior observations on the Ca(2+) distribution in skin were based on Ca(2+) precipitation followed by electron microscopy, or proton-induced X-ray emission. Neither cellular and/or subcellular localization could be determined through these approaches. In cells in vitro, fluorescent dyes have been used extensively for ratiometric measurements of static and dynamic Ca(2+) concentrations, also assessing organelle Ca(2+) concentrations. For lack of better methods, these findings together build the basis for the current view of the role of Ca(2+) in epidermis, their limitations notwithstanding. Here we report a method using Calcium Green 5N as the calcium sensor and the phasor-plot approach to separate raw lifetime components. Thus, fluorescence lifetime imaging (FLIM) enables us to quantitatively assess and visualize dynamic changes of Ca(2+) at light-microscopic resolution in ex vivo biopsies of unfixed epidermis, in close to in vivo conditions. Comparing undisturbed epidermis with epidermis following a barrier insult revealed major shifts, and more importantly, a mobilization of high amounts of Ca(2+) shortly following barrier disruption, from intracellular stores. These results partially contradict the conventional view, where barrier insults abrogate a Ca(2+) gradient towards the stratum granulosum. Ca(2+) FLIM overcomes prior limitations in the observation of epidermal Ca(2+) dynamics, and will allow further insights into basic epidermal physiology.

Demonstration of substances of innate immunity in the esophageal epithelium of domesticated mammals. Part I--Methods and evaluation of comparative fixation.

The aim of the investigation was to demonstrate that the esophageal epithelium of domesticated mammals exhibits characteristic features of innate immunity. The esophageal samples used were obtained immediately after euthanization from seven species of domesticated mammals of three nutrition groups (herbivores: horse, goat, cattle; omnivores: pig, dog, laboratory rat; carnivores: cat). The experimental basis was immunohistochemistry, which was evaluated in a qualitative and statistically relevant semi-quantitative manner. The first part of the study analyzed the influence of different fixation media on the immunohistochemical reactivities. Two formalin-based routine fixation solutions (Bouin's solution, Ca-acetate formalin) were compared with the recently introduced formalin-free HOPE® fixative. In this context, we clearly demonstrated a diminished immunoreactivity for Ca-formol fixed samples; satisfactory results were obtained, particularly, from samples fixed in Bouin's solution. The HOPE® fixation method offers a relatively cheap alternative, as the antibody amounts can be reduced. An application in routine diagnostic is not advisable, because of several variable parameters. It can be concluded that immunohistochemical results have always to be evaluated and discussed in close relation to the fixation medium used.

Demonstration of substances of innate immunity in the esophageal epithelium of domesticated mammals: Part II--Defence mechanisms, including species comparison.

The second part of our study deals with a comparative evaluation and discussion of the immunohistochemical results that were obtained. The cryoscanning electron microscopy (cryoSEM) observations confirmed a monolayer colonization of the esophageal surface with bacteria and fungi (yeasts); the latter in particular was prominent in the ruminant species studied. We demonstrated the existence of several innate immune parameters, including pathogen recognition receptors (PRRs), such as Toll-like receptor 2, which was primarily expressed in the stratum basale; however, the presence ß-glucan receptors remained inconclusive. Furthermore, the group of cationic antimicrobial peptides (CAPs) was shown, comprising ß-defensins 2 and 3 and cathelicidin. The less keratinized esophageal epithelium of the carnivorous cat was protected by high amounts of CAPs; whereas the more strongly keratinized epithelium of the herbivorous and omnivorous species with its characteristic layer structure exhibited clearly weaker reactions. Moreover, lysozyme could distinctly be demonstrated in the cells of the esophageal epithelium. It can be concluded that a first line of defence mechanisms of the innate immune system contributes to maintaining a microbial homeostasis on the surface of the esophageal epithelium of domesticated mammals. The results are discussed in comparison to findings from studies on the human esophagus.

Effect on mode of death of heart failure treatment started with bisoprolol followed by Enalapril, compared to the opposite order: results of the randomized CIBIS III trial.

BACKGROUND: Mode of death in chronic heart failure (CHF) may be of relevance to choice of therapy for this condition. Sudden death is particularly common in patients with early and/or mild/moderate CHF. ß-Blockade may provide better protection against sudden death than ACE inhibition (ACEI) in this setting. METHODS: We randomized 1010 patients with mild or moderate, stable CHF and left ventricular ejection fraction ≤35%, without ACEI, ß-blocker or angiotensin-receptor-blocker therapy, to either bisoprolol (n = 505) or enalapril (n = 505) for 6 months, followed by their combination for 6-24 months. The two strategies were blindly compared regarding adjudicated mode of death, including sudden death and progressive pump failure death. RESULTS: During the monotherapy phase, 8 of 23 deaths in the bisoprolol-first group were sudden, compared to 16 of 32 in the enalapril-first group: hazard ratio (HR) for sudden death 0.50; 95% confidence interval (CI) 0.21-1.16; P= 0.107. At 1 year, 16 of 42 versus 29 of 60 deaths were sudden: HR 0.54; 95% CI 0.29-1.00; P= 0.049. At study end, 29 of 65 versus 34 of 73 deaths were sudden: HR 0.84; 95% CI 0.51-1.38; P= 0.487. Comparable figures for pump failure death were: monotherapy, 7 of 23 deaths versus 2 of 32: HR 3.43; 95% CI 0.71-16.53; P= 0.124, at 1 year, 13 of 42 versus 5 of 60: HR 2.57; 95% CI 0.92-7.20; P= 0.073, at study end, 17 of 65 versus 7 of 73: HR 2.39; 95% CI 0.99-5.75; P= 0.053. There were no significant between-group differences in any other fatal events. CONCLUSION: Initiating therapy with bisoprolol compared to enalapril decreased the risk of sudden death during the first year in this mild systolic CHF cohort. This was somewhat offset by an increase in pump failure deaths in the bisoprolol-first cohort.

A note on the specific cuticle structure of wool hairs in otters (Lutrinae).

The cuticle structure of the wool hairs (secondary hairs) of six otter species was examined by scanning electron microscopy to clarify the specific function of this hair type in the Lutrinae. The species studied were chosen according to the different genera, climatic regions, and degrees of association to water of the Lutrinae. Independent of their preferred habitats, the cuticle of every wool hair examined exhibited in all animals a rather similar shape and arrangement of the scales. This specific adaptive feature allows a flexible interlocking of adjacent wool hairs, which also helps to form thin wool hair bundles that surround small oval shaped spaces. Thus, the trapping of an effective insulating air layer is facilitated and heat loss from the body is reduced.

'Fish matters': the relevance of fish skin biology to investigative dermatology.

Fish skin is a multi-purpose tissue that serves numerous vital functions including chemical and physical protection, sensory activity, behavioural purposes or hormone metabolism. Further, it is an important first-line defense system against pathogens, as fish are continuously exposed to multiple microbial challenges in their aquatic habitat. Fish skin excels in highly developed antimicrobial features, many of which have been preserved throughout evolution, and infection defense principles employed by piscine skin are still operative in human skin. This review argues that it is both rewarding and important for investigative dermatologists to revive their interest in fish skin biology, as it provides insights into numerous fundamental issues that are of major relevance to mammalian skin. The basic molecular insights provided by zebrafish in vivo-genomics for genetic, regeneration and melanoma research, the complex antimicrobial defense systems of fish skin and the molecular controls of melanocyte stem cells are just some of the fascinating examples that illustrate the multiple potential uses of fish skin models in investigative dermatology. We synthesize the essentials of fish skin biology and highlight selected aspects that are of particular comparative interest to basic and clinically applied human skin research.