Alcian Blue Technique Research Articles

Morphogenesis of the ostrich (Struthio camelus) trachea and lung in different embryonic and fetal stages.

The ostrich (Struthio camelus) is an important wild species highlighted in national and international livestock industry. This research was conducted to analyze the development of the ostrich respiratory system during fetal and embryonic stages. A total of 50 fertile ostrich eggs were collected from commercial farms and then incubated at 36.00 - 37.00 ˚C and 25.00 ± 2.00% humidity for 40 days. Sections were taken on days 13, 22, 26, 30, 36, and 42 of incubation from the lung and the cranial, middle, and caudal parts of the neck after decapitation of ostrich embryos and blood drainage. After fixation, processing, blocking, and sectioning, all samples were stained by Hematoxylin and Eosin, Alcian Blue (AB), Van Gieson, and Periodic acid-Schiff (PAS) techniques. It was concluded that the trachea in the 13-day-old embryo and goblet cells (PAS-positive and AB-positive) had incomplete rings of hyaline cartilage and differentiation of mesenchymal to the loose connective tissue. The bronchial stage of the lung was observed in the 22-day-old embryo, pseudoglandular stage in the 26-day-old embryo, and parabrachial and air capillary stage in the 30-day-old embryo. The information obtained from this study will be useful for diagnosing pathologies affecting this vital system and results in improving industrial breeding management.

Mucin levels in glands of the colonic mucosa of rats with diversion colitis subjected to enemas containing sucralfate and n-acetylcysteine alone or in combination.

To evaluate the tissue content of neutral and acidic mucins, sulfomucins and sialomucins in colonic glands devoid of intestinal transit after enemas containing sucralfate and n-acetylcysteine alone or in combination. Sixty-four rats underwent intestinal transit bypass. A colonic segment was collected to compose the white group (without intervention). After derivation, the animals were divided into two groups according to whether enemas were performed daily for two or four weeks. Each group was subdivided into four subgroups according to the substance used: control group: saline 0.9%; sucralfate group (SCF): SCF 2 g/kg/day; n-acetylcysteine group (NAC): NAC 100 mg/kg/day; and SCF+NAC group: SCF 2 g/kg/day + NAC 100 mg/kg/day.Neutral and acidic mucins were stained by periodic acid-Schiff and alcian-blue techniques, respectively. The distinction between sulfomucins and sialomucin was made by the high alcian-blue iron diamine technique. The content of mucins in the colonic glands was measured by computerized morphometry. The inflammatory score was assessed using a validated scale. The results between the groups were compared by the Mann-Whitney's test, while the variation according to time by the Kruskal-Wallis' test (Dunn's post-test). A significance level of 5% was adopted. There was reduction in the inflammatory score regardless of the application of isolated or associated substances. Intervention with SCF+NAC increased the content of all mucin subtypes regardless of intervention time. The application of SCF+NAC reduced the inflammatory process of the colonic mucosa and increased the content of different types of mucins in the colonic glands of segments excluded from fecal transit.

Evidence that the myocardium is a continuous helical muscle with one insertion

Introduction and objectivesAnatomically, the heart cannot be suspended in the thorax without structural continuity of the heart muscle to pump blood at a speed of 200cm/s with an efficacy that allows ejecting 70% of the left ventricular volume with only 12% shortening of its contractile unit, the sarcomere. The aim of this study was to investigate a) whether the myocardium is a single, continuous, helical muscle; b) the origin and end of the myocardial muscle; and c) the sliding movement between the myocardial segments during ventricular torsion–detorsion. This forces us to think of an antifriction mechanism. MethodsA total of 18 hearts (10 bovine and 8 human) were used for anatomical and histological studies. Histological an histochemical analysis of the samples were performed, using the Alcian Blue technique to confirm hyaluronic acid and its antifriction mechanism. ResultsThe myocardium could be defined as a single, continuous, and helical muscle that is inserted at its origin and end, according to the specimens analyzed, to an osseus-chondroid-tendinous nucleus called cardiac fulcrum, thus limiting the 2 ventricular chambers. Hyaluronic acid was found in the excision planes between the myocardial bundles. ConclusionsThe myocardium in its longitudinal continuity adopts a spiraling spatial configuration, inserted at its end to a nucleus called cardiac fulcrum. The Thebesian veins and hyaluronic acid could have an essential role in the antifriction mechanism, due to the resistance between the sliding myocardial muscle surfaces in their torsion (systole) and detorsion (suction) motions.

Development of a new decellularization protocol for the whole porcine heart

Cardiovascular diseases are the leading cause of death in many countries. Advances in technology have been promoted in this regard, especially in tissue engineering, to meet the need for tissue or organ grafts. In this way, the porcine model has been used due to its morphophysiological similarity between the human species, mainly regarding the cardiovascular system. Tissue engineering is employed using biological scaffolds that are currently derived from porcine. These scaffolds are produced by decellularization, a process to remove cells aiming to maintain only its three-dimensional structure, formed by extracellular matrix (ECM). Its main objective is to produce organs through recellularized scaffolds that could eventually substitute the ones with impaired functions. In this way, the present study aimed to establish a new protocol for porcine heart decellularization with potential application on tissue engineering. A porcine heart aorta was cannulated with a silicon tube, and the organ was washed in 0.1% phosphate-buffered saline through a peristaltic pump (Harvard Peristaltic Pump - Harvard Apparatus). After that, deionized water was introduced in the same system. The decellularization procedure was carried out using ionic and non-ionic detergents, namely 4% sodium dodecyl sulfate (SDS) and 1% Triton X-100, respectively. SDS was perfused through myocardial circulation at 400 mL/min for 24 h for 6 days. Subsequently, the heart was infused with Triton X-100 and washed by PBS and water for 24 h. The heart volume was measured before and after the recellularization. After macroscopic evaluation, the heart samples were processed and stained by Hematoxylin and Eosin, Masson's Trichrome, Weigert-Van Gieson, Alcian Blue, and Pricrosirius Red techniques for microscopic analysis. To observe the cell adhesion, the recellularization was provided in this scaffold, which was analyzed under immunofluorescence and scanning electronic microscopy. The protocol provided cells remotion, with adequate concentration of remaining DNA. ECM components as collagen type I, elastin, and glycosaminoglycans were successfully maintained. The scaffold showed a high cells adherence and proliferation in the recellularization process. According to results, the protocol described in this work preserved the ECM components and the organ architecture, minimizing ECM loss and being possible to state that it is a promising approach to tissue bioengineering. This study provides a protocol for whole porcine heart decellularization, which will ultimately contribute to heart bioengineering and may support further studies on biocompatibility relationship of new cells with recellularized scaffolds.

Cutaneous Squamous Cell Carcinoma with Mucinous Metaplasia in Dogs

Background: Squamous cell carcinoma (SCC) is one of the most common malignant skin tumors in domestic animals. Histologically, they are characterized by a proliferation of neoplastic keratinocytes with varied keratin production. Some SCCs have peculiar histological characteristics that permit them to be classified into uncommon to rare histological subtypes, reported in animals and humans. However, according to the authors' knowledge, the mucin-producing subtype described in humans has not yet been reported in animals. In this study, we report the occurrence of two mucin-producing SCCs in dogs, a histological presentation similar to that seen in cutaneous SCCs with mucinous metaplasia in humans.Cases: Two dogs, a 5-year-old Yorkshire female and a 17-year-old Dachshund male, had a skin nodule near the tail and on the right eyelid. The nodules varied from 1 to 5 cm in diameter, were firm and covered with skin and hair. The cut surface was firm and white. Histological findings were compatible with squamous cell carcinoma, characterized by a neoplastic proliferation of keratinocytes originating in the epidermis and infiltrating the dermis. The keratinocytes were arranged in islands and occasional anastomosed cords, supported by a fibrous stroma. The formation of pearls varied from moderate to sparse. The nuclear and cellular pleomorphism was accentuated in case two and moderate in case one. Mitosis figures ranged from two to five in a high magnification field. Within the neoplasm, there were large vacuolated neoplastic cells with slightly fibrillar intracytoplasmic basophilic content. This content has been rarely observed in an extracellular medium. The presence of mucin was confirmed by positive Alcian Blue (AA) staining. In immunohistochemistry (IHC), tumor cells showed strong immunostaining for pancitokeratin, and in areas with marked mucin deposition, immunostaining was predominantly moderate to weak. No tumor cells were immunostained for CD34 and Blc-2 antibodies. Compared to AA and Harris' hematoxylin, it was possible to demonstrate the presence of mucin in the cytoplasm of neoplastic keratinocytes using IHC. No vascular or lymphatic invasion by neoplastic cells was observed. The average cell proliferation index assessed by counting the nucleolar argyrophilic organizing regions (AgNOR) was 3.4 in case 1 and 4.5 in case 2.Discussion: Although the SCC routinely does not present a diagnostic challenge in veterinary practice, the histological presentation of the reported cases does not fit the current classification available in veterinary medicine. The histological presentation observed in these two dogs is similar to that described for cutaneous SCCs with mucinous metaplasia in humans, and so far not described in animals. The observation of intracytoplasmic mucin in humans is an essential finding for the diagnosis of SCC with mucin metaplasia. In the present cases, we observed a slightly basophilic amorphous substance in the cytoplasm of proliferated neoplastic keratinocytes, which stained strongly in blue when applied the Alcian Blue (AA) histochemical technique. This observation became more evident when using IHC counterstained with AA and Harris hematoxylin. In the histological analysis, the absence of an adenoid growth pattern or glandular formation amid neoplastic proliferation ruled out the possibility of a mucinous adenocarcinoma or a mucoepidermoid carcinoma. In addition, we could rule out a follicular neoplasia, including keratinizing infundibular acanthoma due to the absence of a central pore and the absence of immunostaining for CD34 and Bcl-2. These immunohistochemical findings, together with histological findings, reinforce the diagnosis of SCC with mucinous metaplasia in our dogs.

A Study of Histochemical Characteristics of Mucosubstances in Normal Endocervical Glands

Background: The apical epithelial surfaces of mammalian respiratory, gastrointestinal, and reproductive tracts are coated by mucus, a mixture of water, ions, glycoproteins, proteins, and lipids. Mucins are mainly of two types- Neutral and Acidic. Mucus plays an important role in reproductive function and defence of the female reproductive tract. Objectives: To know the mucin distribution in normal endocervical glands. Methodology: This is a retrospective, observational , analytical, case control study aimed to evaluate mucin histochemical pattern in normal endocervical glands. Ten histologically proven blocks of normal endocervical glands were taken. Tissue sections were stained by, PAS, PAS-diastase, Phenylhydrazine-PAS, Alcian blue pH 2.5, Alcian Blue pH 1, combined Alcian blue-PAS, Aldehyde fuschin and combined Aldehyde fuchsin Alcian blue techniques. Results: Results were tabulated according to colour intensity into different grades ranging from + to ++++. Regarding mucin histochemistry of normal endocervical glands; the epithelium and glands show mixture of both neutral and acidic mucins. Acidic mucins are more in amount than neutral. In acidic mucins, sialomucins are predominantly seen. Conclusion: Mucin histochemical patterns serve as valuable, cost-effective tool for diagnosis in histopathology and for the researchers in histology, where a slight change in the mucin pattern may help in the early diagnosis of the disease process.

Superficial acral fibromyxoma of horn : A rare neoplasm and histomorphological entity in an Indian bullock

A four year non-descript bullock was presented to Veterinary Clinical Complex, Veterinary College and Research Institute, Orathanadu, TANUVAS, Tamilnadu with a history of slow growing mass near the base of the left horn. On examination, a protruding, whitish grey, smooth and soft, unlobulated mass was noticed on the body of the left horn. Biopsy specimen was collected and preserved. Histologically, the tissue consisted of mixed population of repetitively arranged fibroblast in whorls with prominent nuclei and stellate cells with cytoplasmic tails. Abundant collagenous and mucinous stroma were present. Abnormal mitotic figures were absent. On special staining, collagenous stroma was more prominent in Masson’s trichrome and blue stained mucinous stroma was seen in Alcian blue technique. Based on the histological features the case was diagnosed as acral fibromyxoma. The case describes the first incidence of fibromyxoma in horn of Indian cattle.

Evaluation of two fixation techniques for direct observation of biofilm formation of Bacillus subtilis in situ, on Congo red agar, using scanning electron microscopy.

Background and Aim:Direct observation, scanning electron microscopy (SEM) is a common method used for the observations of biofilms. N-(3-dimethylaminopropyl)-N’-ethylcarbodiimide) (EDC) fixation method has proven to be a valuable fixation method in the observation of these biofilms. Still, it entails a method of biofilm fixation that can damage slim structures, leading to the impossible observation of biofilm development. In contrast, alcian blue and lysine (ABL) fixation technique appears more glycocalyx of biofilm, fully preserved samples, which may provide much insight into the development of B. subtilis biofilms.Materials and Methods:Here, the evaluation of the fixation of ABL technique for the study of B. subtilis biofilms was carried out in situ, on Congo red agar. In doing so, the comparison to commonly use conventional EDC technique for sample fixation, and observation was carried out. Observations were based on SEM over 30 samples.Results:Overall, ABL technique provided excellent observation of biofilms formed in situ, on Congo red agar, and revealed slime structures, which have not been observed, much in standard EDC fixation or earlier in other studies of these biofilms in B. subtilis.Conclusion:This study reported the appropriate use of ABL in the fixation technique for the preservation of biofilm of B. subtilis.

Hystometric evaluation of nickel chronic exposure effects on large instestine of adult Wistar male rats

The ingestion of considerable amounts of water or food contaminated with nickel can be very toxic. The present work was conducted aiming to evaluate the effects of nickel exposures on ascending colon of adult Wistar male rats at hystometric level. We used 12 animals that were divided in a control (ingested uncontaminated water) and a nickel-contaminated (i.e., 25 mg de nickel/L of water) groups. Nickel chloride was offered in declorinated water and the experiment had a 56 days exposure period. A portion of the ascending colon was removed of the animals and subjected to hystological labelling processes using blue toluidin (for general hystometric description), Alcian Blue (AB, for acid mucins) and periodic acid-Schiff (PAS) technique (for neutral mucins). The potential differences between groups were desgined by applying the Whitney test and t test (p < 0.05). The crypts were smaller for the nickel-contaminated group, even though these organism exhibited broader and higher crypts. Nickel-contaminated animals exhibited a smaller amount of calyceform cells with AB and PAS positive reactions as well as a less mucus quantities when compared with nickel-uncontaminated animals. Such reductions on the amount of calyceform cells with AB and PAS positive reactions may be related wiht the shallower crypts, which possibly reduced the synthesis and secretion of mucins, compromissing the functional aspects (e.g., lubrification and intestinal mucosa protection) of the nickel-contaminated large intestines. Interestingly, the wider and higher crypts and higher epithelium collumn on the nickel-contaminated animals may represent a relevant trade-off for the intestinal mucosa protection.

Influence of the application of enemas with oily extract of curcumin (Curcuma longa) on the tissue content of neutral and acidic mucins in the colonic mucosa without fecal stream

PurposeTo verify if the application of enemas containing oily extracts of curcumin preserves the tissue content of mucins in the glands of the colonic mucosa without fecal stream. MethodThirty-six Wistar rats were submitted to diversion of the fecal stream by proximal colostomy and distal mucous fistula. The animals were subdivided into three groups, and accordingly received enemas with saline and oily extract of curcumin at concentrations of 50mg/kg/day or 200mg/kg/day. After two or four weeks of intervention, the irrigated colic segments were removed. Neutral and acidic mucins were identified by Periodic-acid Schiff and Alcian-Blue techniques, respectively. The content of both mucin subtypes was measured by computerized morphometry. Mann–Whitney test was used to analyze the results, adopting a significance level of 5% (p≤0.05). ResultsThere was an increase in the tissue content of neutral mucins in animals treated with curcumin at a concentration of 50mg/kg/day for four weeks, whereas in the group treated with 200mg/kg/day there was an increase independent of the time of intervention. The content of acidic mucins increased in animals treated with 200mg/kg/day regardless of the intervention time, whereas in those treated with 50mg/kg/day an increase was observed only after four weeks. ConclusionEnemas with curcumin preserve the content of neutral and acidic mucins in the colonic epithelium without fecal stream.

Effect of Moringa oleifera leaf powder supplementation on growth performance and intestinal morphology in broiler chickens.

Due to public health hazards of subtherapeutic use of antibiotics in poultry feed, there is a need for alternatives. Moringa oleifera is a phytobiotic, which is reported to possess antimicrobial and immuno-modulatory properties. This study aimed to investigate effects of M.oleifera leaf powder (MOLP) supplementation on growth performance and intestinal microarchitecture of broiler chickens. Day-old broiler chicks (n=100) were randomly divided into five groups with four replicates each having five birds. Birds were fed a corn-based basal diet or the same diet supplemented with 0.6, 0.9, 1.2 and 1.5% MOLP. Body weight and feed conversion ratio were recorded on weekly basis. On day 35, two birds per replicate were selected to collect samples of duodenum, jejunum, ileum, caecal tonsils and bursa of Fabricius. Tissue samples were stained with haematoxylin and eosin or combined alcian blue and PAS technique. Supplementation of MOLP had no effect on feed intake, feed conversion ratio, bursa weight and intra-epithelial lymphocytes (IEL) count. Birds fed with 1.2% MOLP had higher (p<0.05) body weight, length and empty weight of small intestine, and weight of empty and filled ceca. Villus height (duodenum, jejunum, ileum), villus surface area (duodenum) and villus height:crypt depth (ileum) were higher (p<0.05) in 1.2% MOLP group compared with non-supplemented group. In duodenum, total goblet cell count was higher (p<0.05) in all the supplemented groups when compared with non-supplemented birds. Acidic mucin count was higher (p<0.05) in duodenum, jejunum and ileum of 1.2% MOLP group compared with the control group. Bursal follicle count was higher (p<0.05) in 1.2% MOLP group compared with non-supplemented group. In conclusion, dietary supplementation of 1.2% MOLP modulated intestinal microarchitecture and acidic mucin production without any apparent effect on growth.

Histological and histochemical characterization on stomach of Mystus cavasius (Hamilton), Oreochromis niloticus (Linnaeus) and Gudusia chapra (Hamilton): Comparative study

The histological features and histochemical characterization of the stomach were investigated in Mystus cavasius (Hamilton), Oreochromis niloticus (Linnaeus) and Gudusia chapra (Hamilton) having different feeding habits. Histologically the stomach of all the three fishes was made up of mucosa, submucosa, muscularis and serosa. The mucosa of superficial epithelium consists of a single layer of compactly arranged columnar epithelial cells. Prominent gastric glands are present in M. cavasius whereas in G. chapra the gastric glands are totally absent in the gizzard like stomach. However, in O. niloticus tubular gastric glands are present in the glandular epithelium of caecal like stomach. The distribution and chemical nature of mucopolysaccharides in the aforementioned fishes were studied histochemically by employing Periodic Acid Schiff’s in combination with the Alcian Blue (PAS–AB) technique. Columnar epithelial cells lining the mucosa of the stomach including mucosal border were provided with exclusively neutral mucin which was probably involved in the protective functions against acid and enzymes. The different intensities of reaction of Best Carmine (BC) for glycogen in the epithelial lining and gastric glands of the stomach of the aforesaid three fish species under study were discussed. The intense reaction for protein and tryptophan was noticed in the gastric epithelium and gastric glands of M. cavasius probably due to accumulation of zymogen granules in the gastric glands. On the contrary, moderate reaction for protein and tryptophan was associated with the epithelial cells and gastric glands of O. niloticus and G. chapra. The cytoarchitecture and different degrees of localization of mucopolysaccharides, glycogen, protein and tryptophan in the stomach of M. cavasius, O. niloticus and G. chapra were correlated with the functional significance of the region concerned.

Harderian gland in Canadian ostrich (Struthio camelus): a morphological and histochemical study.

Heads of ten healthy adult ostrich obtained from slaughter house were the constituted materials of the study. The Harderian gland (HG) was dissected out, and all of the gross morphometrical parameters including length, width and thickness as well as weight of left and right glands were recorded. Tissue sections were stained, using haematoxylin and eosin, Masson trichrome, periodic acid-Schiff and Alcian blue (pH 2.5) techniques. In ostrich, HG was an orbital organ located ventromedially around the posterior part of the eyeball. It was an oval flatted shape, light pink colour with irregular outline and was pointed in the dorsal end. Its mean length was 35.30±2.84mm and 35.55±3.58mm in left and right sides, respectively, and mean width 15.30±1.20mm and 15.65±1.18mm in left and right sides, respectively. There was no significant difference between length, thickness, weight and width of left and right glands. Histological results showed that the glandular epithelium was multilobular and compound tubuloalveolar. The gland was surrounded by a connective tissue capsule, and the epithelium was lined by simple columnar epithelial cells of varying height. The secretion of HG was mucous and the secretion type was apocrine. Mucosubstance analysis revealed that secretory units contained acidic and neutral glycoproteins. The granules within the epithelial cells lining the intralobular and inter-lobular excretory ducts of the gland were positive for periodic acid-Schiff and Alcian blue (pH 2.5).

The study of mucin histochemistry in benign and malignant lesions of prostate

Objective: To evaluate the usefulness of mucin stains in differentiating benign and malignant lesions of prostate. Materials and Methods: Sections were obtained from archival paraffin blocks which included randomly selected 70 cases of benign hyperplasia and 30 cases of carcinoma prostate. After confirming the diagnosis, sections were stained for Periodic Acid Schiff (PAS) to study neutral mucins, Alcian blue (2.5 pH) to study acidic mucins and combined Alcian blue - PAS to study the mucin character. Results: Benign hyperplasia of prostate showed positivity for neutral mucins (98.57%) but not for acidic mucins, whereas prostatic carcinomas showed positivity for acidic mucins (46.66%) in addition to the positivity for neutral mucins (56.66%). All the cases of low grade prostatic carcinomas showed positivity for acidic mucins but none of the high grade carcinomas showed positivity for the same. Conclusion: Positivity for acidic mucins with Alcian Blue (2.5 pH) technique can be used to differentiate well differentiated adenocarcinomas of prostate from benign hyperplasia especially in those cases where prostatic lesion is a questionable malignancy either because it is so well differentiated histologically or have altered architecture so as to make it cytologically un diagnosable (P = 0.001).

Parahippocampal corpora amylacea and neuronal lipofuscin in human aging

Abstract The aim of this research was to quantify the number of corpora amylacea and lipofuscin-bearing neurons in the parahippocampal region of the brain. Right parahippocampal gyrus specimens of 30 cadavers were used as material for histological and morphometric analyses. A combined Alcian Blue and Periodic Acid-Schiff technique was used for identification and quantification of corpora amylacea and lipofuscin-bearing neurons. Immunohistochemistry was performed using S100 polyclonal, neuron-specific enolase and glial fibrillary acidic protein monoclonal antibodies for differentiation of corpora amylacea and other spherical inclusions of the aging brain. Cluster analysis of obtained data showed the presence of three age groups (median age: I = 41.5, II = 68, III = 71.5). The second group was characterized by a significantly higher numerical density of subcortical corpora amylacea and number of lipofuscin-bearing neurons than other two groups. Values of the latter cited parameters in the third group were insignificantly higher than the first younger group. Linear regression showed that number of parahippocampal lipofuscin-bearing neurons significantly predicts numerical density of subcortical corpora amylacea. The above results suggest that more numerous parahippocampal region corpora amylacea and lipofuscin-bearing neurons in some older cases might represent signs of its’ neurons quantitatively-altered metabolism.

Effect of dietary supplementation of prebiotics and probiotics on intestinal microarchitecture in broilers reared under cyclic heat stress

This study was designed to evaluate the effect of dietary supplementation of prebiotics, mannanoligosaccharides (MOS) and/or probiotics (LBP) on intraepithelial lymphocytes (IEL) count, goblet cells (GC) count and differentiation and intestinal micro-architecture in broilers reared under cyclic heat stress. Day-old broilers (n = 250) were randomly divided into five groups. Fifty birds were reared within the thermoneutral zone (TNZ). Remaining birds were subjected to cyclic heat stress from day 21 to 42 (35° C, 75% RH, 8 h/d). The birds were fed corn-soy-based basal diet or the same diet supplemented with 0.5% MOS (HS-MOS), or 0.1% LBP (HS-LBP), or their combination (HS-SYN). The birds were slaughtered on day 42. Tissue samples were collected from mid-duodenum, jejunum and ileum, and stained with haematoxylin and eosin or combined Alcian blue and PAS technique. All differences were considered significant at p < 0.05. The IEL count increased in all intestinal segments of the HS group compared with the TNZ group and decreased in all supplemented groups compared with the HS group. Compared with the TNZ, heat stress reduced villus height, crypt depth and surface area in duodenum and ileum, and increased crypt depth in ileum. Villus width decreased in duodenum and jejunum compared with the TNZ group. Supplementation of LBP, MOS and SYN reversed all these changes in duodenum, while only increased villus height and surface area in ileum. In jejunum, the villus height and surface area increased with HS-LBP, and crypt depth increased with HS-MOS. The number of GC containing acid mucins (duodenum and ileum) and mixed mucins (ileum) were increased in the HS compared with the TNZ. Supplementation of MOS, LBP and SYN maintained the enhanced activity of goblet cells. In conclusion, dietary supplementation of MOS and/or LBP may be helpful in alleviating some of the detrimental effects of heat stress on microstructure of the broiler gut.

Decorin content and near infrared spectroscopy analysis of dried collagenous biomaterial samples.

The efficient removal of proteoglycans, such as decorin, from the hide when processing it to leather by traditional means is generally acceptable and beneficial for leather quality, especially for softness and flexibility. A patented waterless or acetone dehydration method that can generate a product similar to leather called Dried Collagenous Biomaterial (known as BCD) was developed but has no effect on decorin removal efficiency. The Alcian Blue colorimetric technique was used to assay the sulfated glycosaminoglycan (sGAG) portion of decorin. The corresponding residual decorin content was correlated to the mechanical properties of the BCD samples and was comparable to the control leather made traditionally. The waterless dehydration and instantaneous chrome tanning process is a good eco-friendly alternative to transforming hides to leather because no additional effects were observed after examination using NIR spectroscopy and additional chemometric analysis.

Effect of polymerized‐type I collagen in knee osteoarthritis. I. In vitro study

Polymerized-type I collagen (polymerized-collagen) is a down-regulator of inflammation and a tissue regenerator biodrug. The aim of this study was to evaluate its effect in co-cultures of cartilage and synovial tissue from patients with knee osteoarthritis (OA). Cartilage and synovial tissue from five patients with OA were co-cultured for 7 days in the presence or absence of 1% polymerized-collagen. To determine proteoglycans content, tissues were stained with alcian blue technique. Pro- and anti-inflammatory cytokines [interleukin (IL)-1beta, IL-8, IL-10, IL-12, tumour necrosis factor (TNF)-alpha, and interferon (IFN)-gamma] and tissue inhibitor of metalloproteinase (TIMP)-1 were measured in supernatants by ELISA and results were normalized by total protein concentration. Cartilage oligomeric matrix protein (COMP), type II collagen, TNF-alpha, IL-10 and Ki-67 expression were determined by immunohistochemistry. Polymerized-type I collagen induced an increase of 3- to 6fold cell proliferation (Ki-67), proteoglycans content, and COMP and type II collagen expression, whereas it inhibited IL-1beta and TNF-alpha production. IL-10 levels were up-regulated in treated vs. untreated cultures. No differences were found on IL-8 or TIMP-1 levels in supernatants from polymerized-collagen-treated co-cultures when compared with untreated cultures. IL-12 and IFN-gamma were undetectable. The addition of polymerized-type I collagen to cartilage and synovial tissue co-cultures induced up-regulation of chondrocytes proliferation and cartilage extracellular matrix proteins production (COMP, type II collagen and proteoglycans) as well as an anti-inflammatory cytokine (IL-10) and the down-modulation of pro-inflammatory cytokines (IL-1beta and TNF-alpha). It is possible that this mechanism might contribute to induce tissue regeneration and down-regulation of inflammation in OA.

Influence of Protein Malnutrition on Prenatal Toxicity of Fluoxetine

Background: Protein malnutrition is prevalent in developing countries. Gestational and neonatal malnutrition were considered to contribute to the development of chronic diseases in adulthood. The aim of the present work was to investigate the influence of prenatal protein malnutrition on embryo -fetal toxic effects of the commonly used antidepressant drug Fluoxetine. Methods: The pregnant rats were divided into two sets: normally fed and protein malnourished. Each set was sub- divided into five groups of 10-14 animals each. Starting from 1st day of pregnancy, animals were fed with standard diet (20% casein). Animals specified for protein malnutrition were switched to protein deficient diet (8%casein) from the 7th day of gestation throughout the end of pregnancy. Fluoxetine hydrochloride was administrated orally in the dose levels 2mg and 8mg /kg/day from day 7 to 14 and from day 15 to 20 of gestation. At the 20th day of gestation the outcome of pregnancy was examined immediately for viability and fetal growth parameters, placental weight as well as fetal external anomalies. Hb, RBCs count, total and differential WBCs and platelets counts were estimated. Fetuses from each group were subjected to skeletal examinations using the Alcian blue and Alizarin red technique. Result:The results showed that prenatal protein malnutrition and administration of Fluoxetine in the high used dose level 8mg /kg were found to cause significant decrease in fetal growth parameters, and placental weights increase in resorption of fetuses multiple hematomas. Blood examination of protein malnourished fetuses and groups treated with fluoxetine revealed significant decrease in Hb level; RBCs count & platelets, however, total the leucocytic count was increase. The incidence of skeletal abnormalities was more obvious when fluoxetine was administrated during embryogenesis period.

Influence of Protein Malnutrition on Prenatal Toxicity of Fluoxetine

Background: Protein malnutrition is prevalent in developing countries. Gestational and neonatal malnutrition were considered to contribute to the development of chronic diseases in adulthood. The aim of the present work was to investigate the influence of prenatal protein malnutrition on embryo -fetal toxic effects of the commonly used antidepressant drug Fluoxetine. Methods: The pregnant rats were divided into two sets: normally fed and protein malnourished. Each set was sub- divided into five groups of 10-14 animals each. Starting from 1st day of pregnancy, animals were fed with standard diet (20% casein). Animals specified for protein malnutrition were switched to protein deficient diet (8%casein) from the 7th day of gestation throughout the end of pregnancy. Fluoxetine hydrochloride was administrated orally in the dose levels 2mg and 8mg /kg/day from day 7 to 14 and from day 15 to 20 of gestation. At the 20th day of gestation the outcome of pregnancy was examined immediately for viability and fetal growth parameters, placental weight as well as fetal external anomalies. Hb, RBCs count, total and differential WBCs and platelets counts were estimated. Fetuses from each group were subjected to skeletal examinations using the Alcian blue and Alizarin red technique. Result:The results showed that prenatal protein malnutrition and administration of Fluoxetine in the high used dose level 8mg /kg were found to cause significant decrease in fetal growth parameters, and placental weights increase in resorption of fetuses multiple hematomas. Blood examination of protein malnourished fetuses and groups treated with fluoxetine revealed significant decrease in Hb level; RBCs count & platelets, however, total the leucocytic count was increase. The incidence of skeletal abnormalities was more obvious when fluoxetine was administrated during embryogenesis period.