Acid Mucopolysaccharides Research Articles

Histochemical Studies on the Male Accessory Glands of the Tenebrionid Beetle Blaps polychresta (Coleoptera, Tenebrionidae)

The aim of the presnt study is to describe the male accessory glands of the tenebrionid beetle Blaps polychresta using histochemical methods and to detect the nature of their secretions. In the adult male tenebrionid beetle Blaps polychresta, there is one pair of bean – shaped and other pair of long tubular accessory reproductive glands. Each bean – shaped gland consists of a single layer of secretory columnar epithelium, surrounded by a muscular coat which is highly tracheated and innervated. Each tubular gland consists of one type of secretory columnar epithelium, surrounded by thin muscular coat which is innervated and highly tracheated. The nuclei of secretory cells in both glands are situated in the basal zone. Histochemical tests of the bean – shaped glands indicate that the secretion of these glands is mainly composed of neutral mucopolysaccharide, glycogen, proteins and lipids. Histochemical tests of the tubular glands indicate the presence of neutral mucopolysaccharide and scanty acidic mucopolysaccharide secretion inside the lumen of these glands, the positive reaction for glycogen and proteins in the cells and secretion inside the lumen of these glands indicate the glycoprotein nature of the secretion.

Topical antibiotics for acute rhinosinusitis in children

Rhinitis or nasopharyngitis are the most common manifestations of acute respiratory viral infection (ARVI) in children. Inflammation of the mucous membrane in the nasal cavity develops in more than 70% of ARVI cases, and rhinovirus infection is the most common etiological factor for these conditions. In this article, the authors present a short review of the existing data on the use of combined drugs based on N-acetylcysteine and tuamingheptan in the treatment of acute rhinological pathology in children. The authors present generalized data on the nature of the course of acute rhinitis, as the main manifestation of acute respiratory viral infection in children, and modern views on the tactics of treating this condition. The main theses on symptomatic treatment of nasal obstruction in acute rhinitis using nasal decongestants are presented. However, the use of nasal vasoconstrictor drugs as monotherapy to treat acute rhinitis may be less effective, as a buildup of thick discharge in the nasal cavity, which a patient is not always able to blow out from the nose (especially in childhood), prevents the drug injected into the nasal cavity from contacting the mucous membrane. The main pathogenetic grounds for the use of mucolytics in the treatment of acute rhinitis and acute rhinosinusitis are presented. The use of N-acetylcysteine-based drugs in otorhinolaryngological practice has been substantiated. N-acetylcysteine is a direct mucolytic agent. Its action is associated with the ability of its free sulfhydryl groups to break intra- and intermolecular disulfide bonds of acidic mucopolysaccharides of a thick discharge. As conclusions, the authors talk about the prospect of using combined drugs based on N-acetylcysteine and tuamingheptan both in the symptomatic treatment of acute rhinitis and as an adjunct treatment for acute rhinosinusitis in children.

Cadmium Toxicity on Chondrocytes and the Palliative Effects of 1α, 25-Dihydroxy Vitamin D3 in White Leghorns Chicken's Embryo.

Cadmium (Cd) can causes osteoporosis and joint swelling. However, the mechanism of Cd toxicity in chondrocytes and how to alleviate Cd poisoning to chondrocytes are still unclear. Herein, we evaluated the toxicity of Cd to chicken chondrocytes, and whether vitamin D can relieve the toxicity of Cd to chondrocytes. Primary chondrocytes were collected from knee-joint cartilage of 15-day-old chicken embryos. They were treated with (0, 1, 2, and 4) μM Cd alone, 10−8 M 1α,25-(OH)2D3 alone, or 2 μM Cd combined with 10−8 M 1α,25-(OH)2D3. We found that Cd significantly inhibited Sox9 and ACAN mRNA expression, which are markers for chondrocyte differentiation, downregulated the mitochondrial membrane potential, upregulated the Bax/B-cell lymphoma 2 ratio. Furthermore, Cd significantly promoted matrix metalloproteinase (MMP)-9 expression, thus accelerating the degradation of extracellular matrix. And Cd also inhibited the expression of main macromolecular protein of extracellular matrix, Collagen type IIα1 (COL2A1) and acid mucopolysaccharide. However, 1α,25-(OH)2D3 pretreatment significantly alleviated the toxicity effects of Cd on the differentiation, apoptosis and extracellular matrix gene expression in primary chondrocytes. Conclusively, Cd exposure could inhibited chicken embryo chondrocytes differentiation, extracellular matrix gene expression, and induced chondrocyte apoptosis. However, these toxic effects of Cd are alleviated by the pretreatment of chondrocytes with 1α,25-(OH)2D3.

Usefulness of high-frequency Doppler ultrasound skin thickness measurement for disease staging and assessing treatment response in patients with scleredema: A case-control study

To the Editor: Scleredema, a rare disease characterized by increased collagen and mucin deposits in the dermis, has been associated with diabetes mellitus, monoclonal gammopathy, and infections.1 The thickness of the dermis in the back can even be multiplied 3- to 4-fold, mainly because of acid mucopolysaccharides deposition.1 The course is unpredictable, with surprising abrupt resolutions or dull persistence for years. The diagnosis is often delayed for months or years.2 Increased body volume due to obesity can simulate the skin's stiffness and thickening in patients with scleredema and delay the diagnosis.

Developmental Changes in Sublingual Salivary Gland of Indian Buffalo during Prenatal and Neonatal Life

Background: The sublingual salivary gland being one of the major salivary glands contributes to a substantial amount of saliva secreted into the mouth. Saliva contains water, various enzymes, mucopolysaccharides and lubricating glycoproteins. It has an important role in providing lubrication for eating and vocalization, aid digestion and supply saliva for pH buffering. The study of prenatal development is prerequisite to understand the normal developmental biology of an organ. The documentation of normal foetal growth can serve as a guide for understanding the consequence of harmful influences at various stages of gestation. The present study was aimed at elucidating the histogenesis of sublingual gland of buffalo during prenatal as well as neonatal life.Methods: The study was carried out on 36 buffalo foetuses, during different stages of prenatal development, as well as 6 neonates. The fetuses were categorized into three groups based on their curved crown rump length (CVRL). The ontogenetic events in histogenesis of sublingual salivary gland of foetal and neonatal buffaloes were observed.Result: The primordial anlage of monostomatic and polystomatic parts of sublingual gland was observed at 39th day and 53rd day of development respectively. The primary ducts (interlobular) were first observed at 80th day of development. The lobulation of the gland was started at 107th day of development, whereas the capsule formation was initiated by the aggregation of mesenchymal tissue at 121st day of development. The duct system was completed at 121st day of development. At same day, the terminal tubules attained the structure of the acini. The typical compound tubulo-acinar nature was first observed at 122nd day of development. The gland showed predominantly mucous type of cells at 137th day of development. The myoepithelial cells were first appeared at 138th day of development. The monostomatic and polystomatic parts were clearly distinguishable by connective tissue at 170th day of development. In neonates, the gland was of compound tubulo-acinar nature with a well-defined capsule. Localization of acidic and neutral mucopolysaccharides was observed in mucous and goblet cells. Fine lipid droplets were observed in intralobular as well as interlobular connective tissue, however, phospholipids were found in the cell membrane of secretory acinar cells and ducts.

Macroscopic, Histomorphological and Histochemical Studies on the Kidneys of Guinea Fowl (Numida meleagris)

Background: Kidneys, the paired organ which essentially plays for excreting nitrogenous wastes, excessive water, inorganic salts and toxic substances produced during the process of body metabolism. Maintenance of osmotic regulation and homeostatic fluid balance of the body has also be performed by kidneys. Each of the kidneys filtrate carried to cloaca by respective ureter from where the urine leaves the body. The present macroscopic, histomorphological and histochemical studies undertaken on the Kidneys of Guinea fowl (Numida meleagris) is nothing but the attempt to widen our thrust area for that bird which is nonexistent as on documentation.Methods: The present studies were carried upon 12 pairs of in-vitro kidney samples in Guinea fowl. Gross morphology and morphometrical observations were carried on fresh specimens. Histological studies were carried on fixed samples those were processed for dehydration, clearing, embedding into paraffin and sections of 5-6 μ thickness were obtained. Different staining procedure as to Haematoxylin and Eosin, Silver orcein, Aniline blue and Weigert’s staining methods were employed for different histological components. The histochemical and histoenzymic studies were also accomplished with Periodic acid Schiff‘s (PAS), Alcian blue methods (pH-2.5) and Gomori’s cobalt method for Alkaline phosphatase and Acid phosphatase. The Succinic dehydrogenase enzyme was also estimated on cryostat sections.Result: The observations on kidneys of Guinea fowl were accomplished after macroscopic, histomorphological and histochemical means. This reddish-brown colored paired organ of kidneys was retroperitoneally placed in synsacral fossa and had incomplete division of three parts. The wrapping material of capsule was the dense connective tissue. The outer cortex and inner medulla had different parts of nephrons. Two types of nephrons viz. mammalian type (with Henle loop) and reptilian type (without Henle loop) were observed. Collagen, elastic and reticular fibers were there in different components of the kidney. Histochemically it has determined the secretion of neutral as well as acidic mucopolysaccharide substances. PAS activity was observed at mesangial cells and basement membrane of DCT. The Alkaline phosphatase showed the positive activity in capsule, brush border of proximal and distal convoluted tubules. The Acid phosphatases were showing intense activity in parietal and visceral layer of renal corpuscles, lumen and basement membrane of PCT. The intense activity of Succinic dehydrogenase showed at PCT, DCT, collecting tubule whereas renal corpuscles showed feeble activity.

ПРИМЕНЕНИЕ ПРЕПАРАТОВ ГИАЛУРОНОВОЙ КИСЛОТЫ В ЛЕЧЕНИИ ОБШИРНЫХ ДЕФЕКТОВ КОЖИ У ПАЦИЕНТОВ, СТРАДАЮЩИХ ДЕСТРУКТИВНЫМИ ФОРМАМИ РОЖИСТОГО ВОСПАЛЕНИЯ НИЖНИХ КОНЕЧНОСТЕЙ

and often requires surgical treatment. Due to the more common development of necrotic forms of erysipelas in patients with dysfunction of local reparative systems, there is a need to stimulate repair to eliminate defects, one of the variants of which may be hyaluronic acid (HA). Hyaluronic acid, which is an acidic mucopolysaccharide by chemical nature, is now widely used in medical practice. This is due to the anti-inflammatory effect, the ability to stimulate the processes of proliferation and epithelialization. We studied the effect of HA in 31 patients with destructive forms of erysipelas in comparison with 32 patients who received standard of care; the results obtained allow us to speak, at least, of numerical trends in favor of HA. Further research is required to evaluate the effect.

Glycan expression as a tool for a deeper understanding of a reproductive gland in a skate of economic importance.

Atlantoraja platana is an oviparous skate endemic to the south-west Atlantic Ocean, and is one of the skate species most exploited by local industrial bottom trawl fisheries. Oviparous elasmobranchs encapsulate their eggs in complex egg cases produced by the oviductal gland (OG). This organ is exclusively present in these fishes and comprises four distinct zones: club, baffle, papillary and terminal. The relative size and structural complexity of these zones correlate with mode of reproduction. Glycans are known to play major roles in reproduction so their distribution in each zone of the OG could explain the functional multiplicity of the gland in skates, but this topic has not been previously investigated. In this study, morphological, histochemical and lectin-histochemical analysis revealed various novel aspects of A. platana's OG. The club, papillary and terminal zones positively stained for periodic acid Schiff's reagent (PAS) and Alcian Blue (AB), indicating the presence of neutral and acid mucopolysaccharides. However, the buffle zone was negative for PAS and AB stains, but was positive for all the lectins used. Each zone of the OG had a characteristic pattern of glycan expression. Finally, we confirmed the presence of sperm but not sperm storage. This is the first lectin-histochemical study of the OG in chondrichtyan fish and it has proven to be an important tool to understand some of the mechanisms of fertility and reproductive success in economic important species such as A. platana.

A novel method for live imaging of human airway cilia using wheat germ agglutinin

Multiciliated epithelial cells in the airway are essential for mucociliary clearance. Their function relies on coordinated, metachronal and directional ciliary beating, appropriate mucus secretion and airway surface hydration. However, current conventional methods for observing human airway ciliary movement require ciliated cells to be detached from airway tissues. Determining the directionality of cilia is difficult. We developed a novel method to stain airway epithelial cilia to observe their movement without releasing ciliated cells. Human tracheae were obtained from patients (n = 13) who underwent laryngectomies to treat malignancies or swallowing disorders. The tracheae were treated with fluorescently labeled wheat germ agglutinin, which interacts with the acidic mucopolysaccharides present on the cilia. Epithelial surfaces were observed using an epi-fluorescence microscope equipped with a water-immersion objective lens and a high-speed camera. Ciliary movement was observable at 125 fps (13/13 samples). Ciliated cells in close proximity mostly exhibited well-coordinated ciliary beats with similar directionalities. These findings indicated that wheat germ agglutinin renders ciliary beats visible, which is valuable for observing human airway ciliary movements in situ.

Internal Anatomy and Biochemical Content of the Cortez Geoduck Panopea globosa during Larval Development

Histological, histochemical, and biochemical changes were assessed in the Cortez geoduck Panopea globosa, during larval development to identify the main energy storage sites and biochemical patterns. Histological sections were stained with polychromic dye, whereas histochemical sections were stained for triglycerides and phospholipids (Sudan black) and for carbohydrates (Alcian blue). Spectrophotometric quantitation of proteins, lipids, and carbohydrates was carried out in 5–35 mg dry weight (DW) samples. Larvae of Panopea globosa showed the typical linear shell growth rate (21 µm d–1) and allometric, power-type relationships between shell length and DW (exponent b = 2.73) or ash-free DW (b = 2.78). Early straight-hinged larvae exhibited a rudimentary digestive system. The crystalline style, intestine, and digestive gland were conspicuous when larvae reached the early umbonate stage, whereas the gills, foot, byssal gland, and adductor muscles were first observed in late umboned larvae. Triglycerides and carbohydrates were mainly stored as droplets in the digestive gland, and their size increased with increasing larval age. Acid mucopolysaccharides were also secreted by the byssal gland. Carbohydrate content was substantially higher than triglycerides in pediveliger larvae. These finding suggests that P. globosa larvae mainly rely on a carbohydrate-based metabolism, both as a biochemical component of the larval–postlarval byssus and as an endogenous energy source during metamorphosis.

Two enigmas may solve each other: the oocyte coat and atresia in the common cockle, Cerastoderma edule (Linnaeus, 1758)

Two co-occurring, enigmatic aspects of bivalve reproduction were investigated in the common cockle Cerastoderma edule: the oocyte coat and oocyte atresia. Qualitative histology and transmission electron microscopy (TEM) of cockles collected on the French Atlantic coast revealed not only the fine structure of the oocyte coat, but also confirmed that it is secreted by the oocyte itself and composed of acid mucopolysaccharides (AMPS), known to be viscous and adhesive. Quantitative histology showed that at the peak of oogenesis, oocyte coats occupy the largest fraction (approx. 40%) of the gonad acinal volume, representing both a significant sacrifice of female gamete capacity, and a non-gamete energetic investment. Potential benefits of the coat include protection from mechanical abrasion, predation, and opportunistic microbes. Atresia (oocyte degeneration) was a known second source of reduced fecundity, with a minimum impact of approximately 50% of the total oocyte volume. It is suggested that this high proportion of atresic oocytes is related to the previously-documented genetic inviability of early post-fertilization life stages. The qualitative histological and TEM observations revealed atresic debris adhering to the exterior surface of the oocyte coats. Such an arrangement would isolate adjacent oocyte coats, enabling the oocytes to be spawned individually, rather than as an egg mass, and therefore to undergo planktonic development and dispersion. Oocyte atresia and the oocyte coat of Cerastoderma edule therefore appear to be linked in the first indication of an adaptive function in bivalves.

Histological and histochemical studies of digestive system in the megrim, Lepidorhombus whiffiagonis (Teleostei: Scophthalmidae)

The digestive system of Lepidorhombus whiffiagonis was studied using histological and histochemical techniques. The wall of oesophagus and stomach fundus consisted of four histological layers: mucosa, submucosa, muscularis and the outer layer. Mucosa, the innermost layer, usually consisted of three different layers, including epithelium, lamina propria and lamina muscularis mucosae. Submucosa of connective tissue with blood vessels and nerve fibres underlined the mucosa and usually protrudes into the mucosal folds except in the gallbladder where it is completely absent. The muscular layer of the digestive system consisted of smooth muscles arranged in longitudinal, circular or oblique layers. Striated fibres were found in the oesophageal wall. The outermost layer comes as tunica adventitia in upper parts and tunica serosa in lower parts of the digestive system. Hepatocytes were surrounded by a mass of fat-storing cells. Serous acini of pancreas were scattered down the length of the stomach. Alcian blue/PAS technique revealed that alkaline and neutral mucopolysaccharides were present in oesophagus while acidic mucopoly-saccharides was found in the stomach and intestines.

PAIN SYNDROMES FORMATION IN DEGENERATIVE SPINE DISEASE

Abstract. Etiological factors of degenerative spine disease and pathogenesis have been considered as a multifactor state. Biochemical and gross pathological changes have been outlined as well. Damage of pulpose disc core is the primary pathological process in degenerative spine disease. Therefore, pathogenetic factors have been defined: reflective, compressive, degenerative, as well as risk factors for development of degenerative and dystrophic changes. Inherent weakness of the connective tissue due to impaired collagen synthesis is significant in development of spine osteochondrosis, which leads to changes in collagen fibers of the fibrous ring, cracks and ruptures in it. Under the influence of various factors (hereditary, mechanical, hormonal, vascular, etc.), depolymerization of acid mucopolysaccharides, proteins, hyaluronic acid of the intervertebral disk pulpal nucleus takes place that leads to dehydration of the disk and reduction of its depreciation. The pathological process in spine osteochondrosis begins with the intervertebral disc, from where it spreads to other structures of the spine. In the pathogenesis of vertebrogenic radiculopathy, biologically active products of depolymerization of the pulp nucleus proteoglycans, as well as autoimmune inflammatory response caused by the pulpous nucleus fragments, which fell into the perineural and epidural space, are crucial. Thus, the established mechanisms and sources of pain in cases of spine osteochondrosis are: irritation of pain receptors, compression of nerve roots and nerve endings, oedema of nerve endings and disc, chemical inflammatory factor. The main factors that cause the pathological process are: autoimmune processes, inflammation, adhesion processes, compression and ischemia. They are closely interrelated and make an ‘endless circle’ in the resistance of pain syndrome in cases of spine osteochondrosis; therewith oedema of nerve endings or root is the main pathogenetic factor. That is why the epidurally administered corticosteroids have a positive result by blocking the receptors of the affected vertebral segments as well as by the action in the region of disco-radicular conflict that leads to reduction or elimination of inflammatory response, which is crucial in pain syndrome development.

Structural and Functional Characterization of the Tongue and Digestive Tract of Psammophis sibilans (Squamata, Lamprophiidae): Adaptive Strategies for Foraging and Feeding Behaviors.

We describe the morphological adaptations of the tongue and gastrointestinal tract of the striped sand snake Psammophis sibilans and discuss their functional importance. Using standard histological, histochemical, and scanning electron microscopy techniques, we analyzed 11 adult snakes of both sexes. Our findings showed that the bifurcated non-papillate tongue exhibited chemoreceptive adaptions to squamate foraging behavior. The lingual apex tapered terminally with sensory spines, and the body of the tongue possesses a characteristic central odor-receptor chamber that might serve to trap and retain scent molecules. Furthermore, the intrinsic musculature showed interwoven and well-developed transverse, vertical and longitudinal muscle fibers that control contraction and retraction during probing and flicking. The esophagus displayed highly folded mucosa lined with columnar epithelium with goblet cells. In contrast, the stomach mucosa formed finger-like gastric rugae, encompassing tubular glands with dorsal gastric pits. The intestine is distinct from other vertebrates in lacking the crypts of Lieberkühn in the tunica mucosa and submucosa. The intestine mucosa is mostly arranged in interdigitating villi oriented perpendicular to the luminal surface. We extrapolated subtle variations for both acid and neutral mucopolysaccharides and glycoproteins localization as well as collagen fibers using histochemical analyses. The elaborate histo-morphological and functional adaptation of the tongue and digestive tract plays a pivotal role in foraging and feeding behavior.

Histological Changes in the Mantle Tissue of the Yesso Scallop Patinopecten yessoensis Shell Infested by Polydora

The Yesso scallop Patinopecten yessoensis is one of the most important aquaculture species in Asian countries because of its high economic value, but it has suffered severe infection by Polydora in recent years, causing great economic losses. Polydora mainly parasitizes the left valves of the Yesso scallop, especially the region around the adductor muscle, badly damaging the shell structure and making the shell particularly fragile. To investigate the response mechanism of the Yesso scallop to Polydora, a histological study was performed on healthy and diseased Yesso scallop mantle tissues, which are thought to be responsible for shell formation. In the present study, separate histological and histochemical changes in different mantle regions were detected for the first time. In the diseased scallops, abundant mucous cells containing acid mucopolysaccharides were found in the epidermis of the outer fold, along with a great increase in cilia on the inner epidermis, and a significant increase in mucous cells secreting acid mucopolysaccharides was observed in the middle fold and the central mantle, suggesting an accelerated process of secretion and transportation of shell substances. These changes in the different mantle regions were probably responsible for the repair of different shell layers. The present study provides valuable information for the function of mantle tissue in shell formation and is helpful for genetic breeding of disease-resistant scallops in the future.

Seasonal variations in histomorphology and histochemistry of the prostate gland of buffalo bulls

The prostate glands of buffalo bulls were collected from an abattoir immediately after slaughter and were fixed in 10% neutral buffered formalin for paraffin block preparation by an acetone-benzene schedule. Paraffin sections of 3-5 μm were obtained on glass slides with a rotary microtome and were stained with various histomorphological and histochemical stains. The distributions of glandular acini and ducts were variable throughout the length of the pelvic urethra. The disseminate part of the prostate gland was derived from the epithelium of the pelvic urethra and was a compound tubulo-alveolar type of gland. The glandular acini comprised three types of cells, i.e. principal (serous cells), mucous, and narrow dark cells. In winter, the lumen of the acini became wider than in summer. The amount of glandular tissue decreased from winter to summer, whereas a reverse pattern was observed in the stroma. The prostatic concretions were larger and centrally placed during winter but became peripheral in summer. The average epithelial height of prostatic acini varied significantly between seasons. The activity of neutral mucopolysaccharides, basic proteins, lipids, and phospholipids was higher in winter than summer, whereas that of acid mucopolysaccharides was higher in summer than winter.

Complimentary Diagnostic Tools for Endometrosis in Biopsies of Mares with Clinical Subfertility

Background: Endometrosis is a multifactorial disease and one of the main causes of infertility in mares, its etiology and pathogenesis are not completely understood. It is defined as peri glandular and/or stromal endometrial fibrosis with glandular alterations. Due to the few clinical symptoms, besides anamnesis and fertility data, endometrosis requires histological confirmation. The histo-morphology and immune histochemical characteristics of the endometrium vary among individuals according to the disease progression. The aim of this research was to combine histology with new immune and histochemical tools for a more precise detection of fibrotic changes of mares with endometrosis.Materials, Methods & Results: The endometrium of forty thoroughbred mares aged 5-18 years, that did not become pregnant during the last two breeding seasons in a Chilean commercial equine breeding center were biopsied. Samples were subjected to conventional histopathology with hematoxylin-eosin as well as to specific histological staining using specific techniques such as Alcian blue and Masson Fontana, aimed to ascertain what types of mucopolysaccharides were present in those samples. In order to have a deeper picture of the progression of the pathology, immune histochemical methods for the detection of vimentin, cytokeratin, progesterone receptor and lymphocyte marker CD3 were used. Finally in order to detect fibrillar collagen we used second harmonic generation (SHG) technique with detects fibrillar collagen without staining, due to intrinsic hyperpolarization ability of this type of collagen, which can be detected by atomic force microscopy. As a result of our research samples were categorized according to the scale of Keeney and Doig into categories I, IIa, IIb and III (45, 42, 7.5 and 5% respectively). These samples also were characterized by the methods listed earlier and a result we found specific staining in 15 samples coming from higher endometrial damage using Masson-Fontana, while acid staining indicative of acid mucopolysaccharides were detected in 97% of the samples. At immunostaining, we found cytokeratin and vimentin differentially expressed as a function of the degree of the lesions. Cytokeratin was detected in glands from healthy mares, but it was almost absent as the injury was increased. Vimentin in turn, was detected in 26 samples with different degree of intensity. In nine animals, vimentin expression was anomalous, and the marker was detected in fibrotic foci concentrically surrounding glands in biopsies of grade I and IIA only. Although progesterone receptor was detected, there was no correlation with endometrosis. Finally, the lymphocyte T specific CD3 marker was positive in 100% of cases analyzed in which moderate lymphocytic infiltration was found by hematoxylin-eosin; however no staining could be detected in mares with more advanced endometrosis. By combining immunofluorescence with the detection of second harmonic it was possible to detect in the same sample two proteins and collagen deposition at the same time, this had not been reported earlier for endometrosis and mares.Discussion: Our results suggest that the combination of the different methods mentioned is useful for validation and further characterization of the routine hematoxylin-eosin, and can be used as a complementary tool for the diagnosis. Of value, it was possible to combine immunofluorescence with SHG in a single sample in situ.

Correlation between smoking and neovascularization in biceps tendinopathy: a functional preoperative and immunohistochemical study.

Aims:The purpose of this study was to investigate whether smoking is associated with neovascularization in the tendinopathy of the long head of the biceps tendon (LHBT).Methods:The study included 40 consecutive patients who underwent arthroscopic biceps tenotomy/tenodesis due to chronic biceps tendinopathy and divided into three groups: (1) non-smokers, (2) former smokers, (3) smokers. LHBT tissue samples were stained with H&E, Alcian blue and Trichrome staining. Immunohistochemical examination was performed using anti-CD31 and anti-CD34. The neovessel density score (NDS) was scored by Bonar criteria.Results:The mean period of smoking was 15.50 years with an average number of 24 cigarettes/day in the former smokers and 21.69 years with an average number of 15 cigarettes/day in the active smokers. The mean NDS was 2.23/3 in non-smokers, whereas it was 1.60/3 in former smokers and 1.31/3 in active smokers. The mean American Shoulder and Elbow Surgeons score equaled 46 in never smoked patients, 43.60 in former smokers, and 41.46 in active smokers. In the patients with smoking history, the disorganized tendinous tissue islands were avascular and composed of compact acidic polysaccharides and mucopolysaccharides. We observed negative correlation between the NDS and the smoking indexes, including cigarettes per day (p = 0.0150), smoking years (p = 0.0140), pack-years (p = 0.0088).Conclusion:In conclusion, the present study revealed that smoking impairs the vascularization of the biceps tendon in chronic tendinopathy. Clinically, we observed a negative correlation between smoking and neovascularization. Furthermore, there was no correlation between neovascularization and functional preoperative status.

Hyaluronidase pharmacological properties and clinical application in ophthalmology

Corneal diseases are the fourth leading cause of blindness in the world, accounting for approximately 5 % of cases. Existing methods of treatment in more than 30 % of cases do not have a full therapeutic effect and when the process is stopped, they end with a persistent violation of corneal transparency, a decrease or complete loss of visual functions. The lack of effective means that evidence-based restore corneal transparency determines the relevance of the search for modern drugs and ways to deliver them, the possibilities of enhancing the therapeutic effect. The possibilities of using hyaluronidase preparations in ophthalmology are considered based on the study of its biological and pharmacological properties. Enzyme catalyzes the breakdown of acid mucopolysaccharides including hyaluronic acid by cleavage of the glycosidic bond β(1→4), the biological effect is determined by the molecular weight of the resulting fragments: high-molecular fragments have antiangiogenic properties, increased ability to bind fibrinogen, anti-inflammatory and immunosuppressive effects, and low-molecular fragments have pro-inflammatory activity and promote angiogenesis. The barrier function of the cornea is provided by the features of its anatomical structure, while the features of its regeneration with the formation of turbidity are accompanied by overexpression and migration to the stroma of cytokines TGF-β and PDGF, activation of myofibroblasts and the formation of a fibroproliferative response. The high anti-inflammatory, immunomodulatory, regenerative and antifibrotic activity of hyaluronidase, the possibility of its effect on a complex pathophysiological cascade of destructive processes and minimization of the scarring process stimulate more extensive experimental and clinical studies on the development of new methods of treating ophthalmic diseases using hyaluronidase drugs.

Influence of germination of wheat grain with selenium sources on the components of protein-carbohydrate complex

The aim of the study is histochemical identification of the components of protein-carbohydrate complex of wheat grains with selenium sources during germination. We formulated a hypothesis about the stimulating effect of 4.4-di [3 (5-methyldipirazolyl] selenid (DMDPS) on the biosynthesis of substances with a bactericidal effect, which include mucoid substances and glycoproteids in the composition of the components of the protein-carbohydrate complex. We confirm the hypothesis on histochemical identification of the components of the protein-carbohydrate complex of wheat grain during germination with sources of selenium. To identify acid mucopolysaccharides, transverse and longitudinal sections of the grain were stained with Alcian blue according to Steedman. Schiff-iodic acid revealed neutral glycoproteins and was clarified in xyllol. The sections were then placed into the balm and viewed in a Zeiss Axioscop 40 FX light microscope using a Levenhuk C 510 NG digital camera. To detect methionine, sections were stained with Schiff’s reagent; to identify cysteine (SH-groups), sections were treated with a 0.05% solution of 2.2-dioxy-6.6-dinaphthyl disulfide. The obtained images were analyzed using Photoshop CS 5 tools. Positive effect of wheat germination with DMDPA on the content of both neutral glycoproteids and mucopolysaccharides, with a direct correlation relationship, and on the content of methionine and cysteine was proved.