Connective Tissue Area Research Articles

Suspected panosteitis in a camel

A 6-month-old male Bactrian camel was examined because of a 3-week history of lameness of the left hind limb. Lameness was initially detected in the left hind limb but resolved and was detected in the right hind limb during treatment. Lameness increased during periods of rapid growth. Radiography revealed multiple small opacities of the medullary cavity of several long bones throughout treatment. Core bone biopsies of lesions in the tibiae revealed lamellar bone with areas of loose connective tissue, osteoblasts in the medullary cavity, and periosteal new bone formation, all which were consistent with panosteitis. Palliative treatment was attempted with epidural and transdermal administration of analgesics. Flunixin meglumine was administered PO, which coincided with an abrupt increase in serum creatinine concentration. Performance of multiple diagnostic bone biopsies led to remission of clinical signs of pain. Panosteitis should be a differential diagnosis for shifting limb lameness in young camels. Bone biopsies can be useful for diagnosis of panosteitis and possible relief of pain associated with the disease. Bactrian camels may be susceptible to the renal toxicity of flunixin meglumine, especially when dehydrated.

Curto período de imobilização provoca alterações morfométricas e mecânicas no músculo de rato

Short-term immobilization causes morphometric and mechanical alterations in rat muscles Objective: to analyze the morphometric and mechanical characteristics of the soleus and gastrocnemius muscles after immobilization in a shortened position. Methods: 20 Wistar rats (250 ± 20g) were divided equally into immobilized and control groups. The left hind limb was immobilized by means of an acrylic resin orthosis, with the ankle joint at maximum plantar flexion. After seven days of immobilization, the muscle mass, number and length of sarcomeres in series, muscle fiber cross-sectional area, density of the intramuscular connective tissue area and tensile strength of the triceps surae muscle were evaluated. The data were analyzed by the ANOVA and Tukey tests (p< 0.05). Results: The immobilized soleus muscle presented changes in all the morphometric variables analyzed, while some of these changes were not observed in the gastrocnemius muscle. Analysis of the traction test showed that the immobilized group presented a 20% decrease in the maximum tensile muscle strength. Conclusion: The results from this study showed that short-term immobilization causes changes to the morphometric parameters of the muscle fibers, with repercussions on muscle mechanics. These results suggest the need for rehabilitation of muscles subjected to immobilization, even if only for a short period, in order to achieve early recovery of normal muscle characteristics.

Increased Cx43 and Angiogenesis in Exercised Mouse Hearts

Several studies focused on the macroscopic architecture of increased cardiac wall induced by exercise training. Our goal was to evaluate myocardiocyte, interstitial and vascular component, and connexin-43 expression in endurance-trained mouse hearts. Sixty-three 10-week-old male Swiss mice were divided into four sedentary groups (C0, C15, C30 and C45) and three groups exercised respectively for 15 (T15-D; running intensity [RI]: 3.18 m/min; running duration [RD]: 75 min/first week and 150 min/second week), 30 (T30-D; RI: 3.96 m/min; RD: 150 min/third week and 225 min/fourth week) and 45 days (T45-D; RI: 3.96 m/min and 4.8 m/min, respectively for the fifth and sixth week; RD: 300 min) on a treadmill. Morphometric analyses were performed to quantify myocardiocyte size and number, and the capillary and interstitial connective tissue (ICT) area. We assessed the expression of ventricle myosin light chain-II, vimentin and connexin-43 by western blot analyses. Our results showed a hypertrophy of the interventricular septum and left ventricle in T30-D and T45-D mice that was not due to variations in myofibrillar content, myocardiocyte size and number, or ICT quantity but to a significant increase in the capillary area. The microvascular remodeling was associated with vimentin increased expression in ICT cells and connexin-43 upregulation. The first phenomenon might be related to an enhanced request of remodeling and growth factors; the second suggests a new role of connexin-43 in cardiac angiogenesis.

Biomechanics and Histology of Intact and Repaired Digital Nerves: An In Vitro Study

To investigate the biomechanical properties of intact and repaired cadaver digital nerves. Ultimate tensile failure strength and stiffness were determined in 67 human cadaver digital nerves. Total nerve area, fascicular area, and nonfascicular (connective tissue) area were determined from the metacarpophalangeal to the distal interphalangeal joint in another 35 axial nerve sections to determine regional anatomic variation. Thirty-eight additional digital nerves were transected, and epineural repairs were performed using simple, interrupted sutures. Suture number (2 vs 4), gauge (8-0 vs 9-0), and purchase length (1 mm vs 2 mm) were used in various combinations, and then the repaired nerves were pulled to failure. The mechanism of repair-site failure was determined for each suture. In situ tension of the intact digital nerves was measured during passive metacarpophalangeal and proximal interphalangeal joint motion in another 19 intact digital nerves. There were no significant differences in failure load or stiffness with respect to the radial or ulnar nerves within a finger or between fingers. The primary tactile side of the finger tended to have a larger diameter digital nerve. Digital nerve failure was more common proximally than distally. Intact digital nerves failed at 6 N with a stiffness of 1 N/mm. Histologic analysis showed that fascicular area and total area decreased from proximal to distal, whereas the nonfascicular-to-fascicular area ratio increased. Four epineural sutures were statistically stronger than 2 sutures. Suture purchase length and gauge did not affect repair strength. The 8-0 nylon sutures failed primarily by cut-out, whereas the 9-0 sutures failed by cut-out or breakage. Repaired nerves failed at 1 to 2 N. Maximal metacarpophalangeal joint hyperextension resulted in 4 N of digital nerve tension. When the metacarpophalangeal joint was not hyperextended, proximal interphalangeal joint motion did not generate tension. Similar to flexor tendons, the number of suture strands crossing the repair site was the most important variable affecting digital nerve repair strength in this cadaveric model.

Control of ham salting by using image segmentation

Curing is one of the most traditional processes in the meat industry, being used in a great variety of products such as cured ham. During the salting process the raw material acquires the curing agents, allowing the safe development of subsequent stages in processing. Digital image analysis has been used in different food research areas. Most of the studies that use image analysis in the evaluation of different aspects of meat products have been carried out mainly on ham, detecting quality problems of the product. However, none of these studies deals with the influence of different components present on the ham surface (fat, connective tissue and lean) and its relationship with mass transfer during ham processing. The aim of this study was the use of image segmentation to quantify the lean, fatty and connective tissue areas on the ham surface and determine the relationship of those areas to salt gain during the salting process. The obtained results show that image segmentation algorithm can be used in combination with other parameters values to predicting ham behaviour during the salting process.

Role of tumour necrosis factor α, but not of cyclo‐oxygenase‐2‐derived eicosanoids, on functional and morphological indices of dystrophic progression in mdx mice: a pharmacological approach

The role of tumour necrosis factor (TNF)-alpha or cyclo-oxygenase-2 (COX-2) eicosanoids in dystrophinopathies has been evaluated by chronically treating (4-8 weeks) adult dystrophic mdx mice with the anti-TNF-alpha etanercept (0.5 mg/kg) or the COX-2 inhibitor meloxicam (0.2 mg/kg). Throughout the treatment period the mdx mice underwent a protocol of exercise on treadmill in order to worsen the pathology progression; gastrocnemious muscles from exercised mdx mice showed an intense staining for TNF-alpha by immunohistochemistry. In vivo, etanercept, but not meloxicam, contrasted the exercise-induced forelimb force drop. Electrophysiological recordings ex vivo, showed that etanercept counteracted the decrease in chloride channel function (gCl), a functional index of myofibre damage, in both diaphragm and extensor digitorum longus (EDL) muscle, meloxicam being effective only in EDL muscle. None of the drugs ameliorated calcium homeostasis detected by electrophysiology and/or spectrofluorimetry. Etanercept, more than meloxicam, effectively reduced plasma creatine kinase (CK). Etanercept-treated muscles showed a reduction of connective tissue area and of pro-fibrotic cytokine TGF-beta1 vs. untreated ones; however, the histological profile was weakly ameliorated. In order to better evaluate the impact of etanercept treatment on histology, a 4-week treatment was performed on 2-week-old mdx mice, so to match the first spontaneous degeneration cycle. The histology profile of gastrocnemious was significantly improved with a reduction of degenerating area; however, CK levels were only slightly lower. The present results support a key role of TNF-alpha, but not of COX-2 products, in different phases of dystrophic progression. Anti-TNF-alpha drugs may be useful in combined therapies for Duchenne patients.

Brachialgia paraesthetica nocturna can be relieved by “wet cupping”—Results of a randomised pilot study

Centuries ago cupping was one of the most used medical therapies worldwide but it is now regarded as an antiquated and unsafe treatment. Nevertheless it is widely used especially in Germany and China. To investigate the effectiveness of "wet cupping" of a defined connective tissue area (over the Musculus trapezius) in patients suffering from brachialgia paresthetica nocturna. Monocenter, randomised, controlled, sequential clinical trial. Section of pain management at the District Hospital of Rüdersdorf, Germany. Brachialgia-patients of both sexes without age restictions were eligible if they suffered from chronical tonsillar irritations and showed pathologic indurations of the connective tissue area. The active group was "wet cupped" once, i.e. the skin first was scarified and then blood was drawn by applying vacuum cupping glasses. The control group was left untreated. Pre- to post-treatment change of brachialgia severeness, calculated from 1-week averages of the means of three subscales (pain, tingling and numbness), each assessed on a 0-10 numeric analogue scale. N=20 patients were randomised (13 women, median age 47 years). Treatment effects can be found in the active (-2.3+/-1.9 score points) but not in the control group (+0.5+/-1.0 points; p=0.002; triangle test). The results are supported by secondary outcome criteria. Adverse events were not documented in any patient. This study suggests short-term effects of a single wet cupping therapy, which remain at least for 1 week. As the trial lacks of an adequate and blinded placebo therapy the findings are potentially biased.

Immunoreactivities to protein gene product 9.5, neurofilament protein and neuron specific enolase in nerves in the oviduct of the sexually immature ostrich, &lt;i&gt;Struthio camelus&lt;/i&gt;

The present study investigated the distribution of nerves in the oviduct of the sexually immature ostrich. The presence of protein gene product 9.5, neurofilament protein and neuron specific enolase nerve fibres were demonstrated in the infundibulum, magnum, isthmus, shell gland and vagina. Nerve fibres containing protein gene product 9.5, neuron specific enolase and neurofilament protein were particularly numerous in the tunica muscularis and intermuscular connective tissue areas of the shell gland and vagina. The presence of a large number of nerves in these oviductal regions is probably important in the coordination of muscle contraction. An interesting finding of the study was the presence of protein gene product 9.5 and neuron specific enolase immunopositive nerve fibres in the walls of blood vessels. It is likely that these nerves are autonomicin nature and play a role in the regulation of blood flow. This study has shown the presence of an extensive neural network in the oviduct of the ostrich. In addition, the results of the investigation have shown that the neuronal markers protein gene product 9.5, neurofilament protein and neuron specific enolase can be used to demonstate nerve fibres in the ostrich.

Efeitos da estimulação elétrica neuromuscular sobre o membro posterior imobilizado de ratos durante 15 dias: análises metabólicas e morfométricas

OBJETIVO: Avaliar o efeito da estimulacao eletrica (EE) sobre o perfil metabolico e morfometrico dos musculos do membro posterior de ratos submetidos a imobilizacao durante 15 dias. METODO: Ratos Wistar foram divididos em 3 grupos (n=5): controle, imobilizado por 15 dias e imobilizado associado a EE por 15 dias. Foram avaliados: reserva de glicogenio (RG) dos musculos soleo (S), extensor longo dos dedos (ELD), gastrocnemio branco (GB), gastrocnemio vermelho (GV) e tibial anterior (TA), alem do peso do soleo, area das fibras e tecido conjuntivo do S. A analise estatistica foi feita pelos testes ANOVA e Kruskal-Wallis (p<0,05). RESULTADOS: A imobilizacao promoveu alteracoes significativas (p<0,05) como: reducao nas RG (S: 44,73%, GB: 47,82%, GV: 46,34%, ELD: 41,66%, TA: 48,38%), no peso (7,2%) e na area das fibras (35%) do S, alem do aumento da densidade do tecido conjuntivo (160%). A EE promoveu aumento significativo (p<0,05) nas RG de todos os musculos imobilizados (S: 90,47%, GB: 62,5%, GV: 95,45%, ELD: 76,19%, TA: 56,25%), no peso (20,94%) e na area das fibras (19,65%) do S e tambem promoveu reducao significativa (15,38%, p<0,05) na densidade do tecido conjuntivo. CONCLUSOES: A EE minimizou a reducao das RG, preveniu a reducao da area das fibras e a proliferacao do tecido conjuntivo nos musculos submetidos a imobilizacao.

Comparison of human tonsillar mast cell localization and ultrastructural observations between IgE-mediated allergic and nonallergic donors

Mast cells (MCs) are important effector cells in mucosal defense and allergic inflammation. Lymphoid tissues such as tonsils and adenoids also have modest numbers of MCs. However, the role of MCs in lymphoid tissues is not well known. In this study, we showed the local distribution of MCs in tonsils from IgE-mediated allergic or nonallergic donors, studied their ultrastructure, and specified their surrounding cell types. Tonsils were obtained from IgE-mediated allergic or nonallergic donors suffering from chronic tonsillitis or hyperplastic tonsils. The localization and distribution of MCs and IgE binding to MCs were determined by immunohistochemistry using antibodies against tryptase, c-kit, and IgE. In addition, mast cell structure was examined using electron microscopy. In both allergic and nonallergic donors, MCs were distributed mainly in the interfollicular area and the perivascular area of connective tissue. We found that the total number of MCs in tonsils was almost the same in both donor groups. However, the number of MCs in the interfollicular area was significantly higher in nonallergic versus allergic donors. Moreover, the electron microscopy revealed that MCs localized in perivascular areas of connective tissue have scroll-type granules, and MCs in the interfollicular areas contained both particle and scroll-type granules. In addition, the MCs that were surrounded by CD4+ lymphocytes in the interfollicular area showed empty granules, whereas MCs in the perivascular area, not surround by CD4+, were intact. This implies that these MCs were degranulated, and this probably was caused by CD4+ cells. Taken together, these findings suggest that tonsillar MCs distribute differently in allergic and nonallergic donors, and that MCs in the interfollicular area might be activated by direct contact with CD4+ T cells.

Assoziationen zwischen chronischer Tonsillenreizung, Verquellungen von Bindegewebszonen und einer Brachialgia paraesthetica nocturna

Projective and reflex zones of inner organs tothe body’s surface are well described but there are only fewclinical studies about abnormal zones. Objective: The aimof our study was to investigate associations between chronicirritation of tonsils, abnormalities of the connective tissueareas over the musculus trapezius and the severity ofbrachialgia paresthetica nocturna. Methods: Cross sectionalstudy in 100 adult pain patients from an outpatient pain department.Patients with tonsillectomy were excluded. Examinationsof tonsils and the connective tissue area were doneseparately by two mutually blinded physicians. Both werealso blinded to the patients’ evaluation of his/her brachialgia.Results: Regardless of the lateralization, the severity ofthe tonsil irritations was correlated with the abnormalities ofthe connective tissue areas (Spearman’s ρ = 0.82; 95% confidenceinterval (CI): 0.74-0.87; p < 0.001) and the severity ofthe brachialgia (ρ = 0.64, CI: 0.50-0.74; p < 0.001), furthermoreindurations of connective tissue areas correlated withthe severity of brachialgia homolaterally (ρ = 0.57; CI:0.42-0.69; p < 0.001). These correlations are considerablyhigher than those of other connective tissue areas. Conclusions:The results support the existence of easy to diagnosereflex zones, at least in a highly selected population of painpatients. They can give plausible hints for naturopathictreatments of brachialgia paresthetica nocturna, i.e. treatmentof the relevant connective tissue zone above the M.trapezius.

Treatment of Class III Furcation Defects With Expanded Polytetrafluoroethylene Membrane Associated or Not With Anorganic Bone Matrix/Synthetic Cell‐Binding Peptide: A Histologic and Histomorphometric Study in Dogs

Up until now, no predictable periodontal regeneration of Class III furcation defects has been demonstrated after treatment with different available techniques. Recently, a bone graft enriched with a peptide was developed and has shown satisfactory results when applied in intrabony defects. The aim of this study was to compare the use of expanded polytetrafluoroethylene (ePTFE) membrane associated (test group) or not (control group) with anorganic bovine-derived bone matrix (ABM)/synthetic peptide, in the treatment of Class III furcation defects in dogs. Six mongrel dogs were used in this study, and the second and fourth mandibular lower premolars were extracted. Class III furcation defects were surgically created in the third premolars and filled with impression material. Afterwards, the defects were surgically assessed for debridement and root planing. Teeth were randomly assigned into test and control groups. The membranes were removed after 4 weeks, and the animals were sacrificed 12 weeks later. Comparisons between groups by the Wilcoxon signed rank test showed no statistically significant differences in the parameters evaluated. In the control group, a new bone area (NBA) of 41.71%+/-24.07%, connective tissue area (CTA) of 36.34%+/-15.50%, and epithelium tissue area (ETA) of 9.39%+/-5.85% were observed. The new cementum extension (NCE) was 24.16%+/-13.18%. The test group presented an NBA of 31.84%+/-12.58%, CTA of 47.72%+/-11.33%, ETA of 9.17%+/-6.81%, and an NCE of 30.13%+/-16.43%. There was no statistically significant difference between the two therapies: ePTFE membrane associated with ABM/synthetic peptide flow or ePTFE membrane only.

Diltiazem did not induce gingival overgrowth in rats: a clinical, histological and histometric analysis

The administration of calcium channel blockers has been associated with gingival overgrowth. However, there are few studies in humans or animals that evaluated the effect of diltiazem on gingival tissues. The present study assessed the influence of diltiazem, at different dosages and treatment duration, on gingival tissues of rats, using clinical, histological and histometric analyses. Eighty young male rats were separated into eight groups according to the dosage and duration of treatment. Rats were treated for 20 or 40 days with a daily subcutaneous injection of 5, 20 or 50 mg/kg of body weight of diltiazem. The results confirmed that diltiazem did not induce gingival overgrowth in rats. For all animals, the evaluation did not show gingival alterations regardless of the dosages and periods of treatment. The histometric analysis showed no significant change in the area of epithelium and connective tissues, although after 40 days of treatment a decrease in the area of connective tissue was observed, without statistically significant difference from control groups. Within the limits of this study, we suggest that diltiazem did not induce gingival overgrowth.

Morphology after synergistic terminal end-to-side nerve graft repair: Investigation in a nonhuman primate model

BACKGROUND: Functional efficacy of synergistic terminal end-to-side nerve graft repair in a median nerve defect model in nonhuman primates was recently published elsewhere. The purpose of this study was to investigate the morphology of the end-to-side repair site and the nerve graft used after synergistic terminal end-to-side nerve graft repair. METHODS: Experiments were carried out in seven adult baboons. Synergistic end-to-side nerve graft repair bridging from the terminal motor branch of deep branch of the ulnar nerve to the thenar motor branch of the median nerve was performed after transection of the median nerve at forearm level. Specimens were harvested three months after surgery. Chromotrope aniline blue was used for staining neural collagenic connective tissue. S100-antibody was used to quantify the number of Schwann cells. Axon counts were performed using NF (neurofilament) antibody staining. To enable profound morphological assessment, specimens were harvested 5 mm proximal to the end-to-side nerve graft repair site (deep branch of ulnar nerve), at the end-to-side nerve graft repair site (proximal graft), and at the distal end-to-end nerve graft suture site. The contralateral terminal deep branch of ulnar nerve served as a control. RESULTS: The area of collagenic connective tissue was 6% larger at the end-to-side repair site and 28% larger at the distal end of the nerve graft. Highest mean values were found at the distal site of the nerve graft. This was due to very strong collagenization of the nerve graft of animal number six. In other words, there was a moderate enhancement of collagenic connective tissue within the nerve graft in six animals. Schwann cell counts showed similar values within all nerve segments. There was no difference when compared to nonoperated control. The number of axons distal to the end-to-side repair site was smaller by 19% on average. Additionally, we found an average 23% decrease of axons at the end-to-end coaptation site compared to data of the deep branch of ulnar nerve. CONCLUSIONS: The results in this morphological study of synergistic end-to-side nerve graft repair at the level of very peripheral terminal motor branches in a nonhuman primate model demonstrate effective distal axonal sprouting and successful Schwann cell survival. As expected, we found a modest increase of neural collagenic connective tissue within the nerve graft in most animals (6 out of 7 animals).

Healing following tooth extraction in cyclosporine-fed rats

Healing after tooth extraction was studied in rats treated with cyclosporine-A (CSA) for four weeks. Sixty male Sprague–Dawley rats were assigned to one of three groups of 20 rats each. The maxillary right molars were extracted from two groups; the third group served as a non-extraction control. The non-extraction group and one extraction group (vehicle control) received the solvent mineral oil daily, and the other extraction group received 15 mg/kg CSA in mineral oil. Five rats from each group were killed 5, 10, 14 and 28 days after extraction and samples analyzed histologically. On days 5 and 10, bone volume was significantly lower and marrow volume significantly higher in both extraction groups than in the non-extraction group. The fractional-formation surfaces were significantly lower in the extraction groups than in the non-extraction group on day 5 only. Osteoid volume was significantly higher in the extraction vehicle control group than in the other two groups on days 10 and 14; however, the osteoid volume was higher in the CSA group than in the other two groups on day 28. On days 14 and 28, bone volume was lower and marrow volume higher in the CSA group than in the extraction vehicle control and non-extraction groups. On day 28, bony surface areas were significantly greater in the CSA group than in the extraction vehicle control and non-extraction groups. Soft-tissue evaluation showed significantly greater epithelial areas, connective tissue areas and total tissue areas in the CSA group than in the extraction vehicle control group on day 28, but not on day 14. These data suggest that CSA may influence healing of both the gingival tissue and the alveolar bony sockets in the tooth-extraction wound. Further detailed study is needed to identify the mechanisms responsible.

Lymphangiosarcoma in a 3.5-year-old Bullmastiff bitch with vaginal prolapse, primary lymph node fibrosis and other congenital defects : clinical communication

Lymphangiosarcoma is an extremely rare tumour in dogs with only 16 cases reported in the literature. Lymphoedema, which may be primary due to defects in the lymphatic system, or secondary to various other pathologies, often precedes malignancy. Of the 16 canine reports, only 1 dog was confirmed as having had prior primary lymphoedema due to aplasia of the popliteal lymph nodes. A case of lymphangiosarcoma is described in a 3.5-year-old purebred, Bullmastiff bitch which presented with vaginal blood 'spotting' for 3 weeks after cessation of oestrus, during which intromission by the male had been unsuccessful. During ovariohysterectomy a large multicystic, proliferative, spongy, fluid-filled, brownish-red mass surrounding the cervix and projecting into the abdominal space was removed with the cervix, and a diagnosis of lymphangiosarcoma made on histological and electron microscopic examination of the tissue. Ultrastructurally, no basement membrane or pericytes were found, only some of the neoplastic endothelial cells were linked by tight junctions while there were gaps between others, and neither micropinocytotic vesicles nor Weibel-Palade bodies occurred in the cells examined. Very few of the endothelial cells lining the many interlinking, tortuous maze of channels, stained slightly positive immunohistochemically for factor VIII-related antigen. The channels were filled mostly with serous fluid, and occasionally mixed leucocytes and some erythrocytes. The endothelium was often associated with underlying blocks of collagenous material, as well as loosely-arranged aggregates of lymphocytes, other mononuclear cells and occasional neutrophils in the connective tissue septae and more prominently perivascularly. The bitch was discharged on antibiotic treatment but returned 2 weeks later with apparent prolapsed vagina which failed to reduce over the next week. Laparotomy revealed the tumour to have spread extensively in the caudal abdomen to involve the broad ligament and the ventral rectal serosa, and the 'prolapsed' tissue was found to be expanded vaginal wall. The bitch was euthanased and necropsied, Histological examination confirmed lymphangiosarcomatous invasion of the submucosal and muscular layers of the retroperitoneal, traumatised, prolapsed part of the vagina, the urethra and the ventral rectal wall. The broad ligament was diffusely invaded with tumour which had proliferated into the caudal abdominal space, and 3 small intra-trabecular foci of tumour were found in the right popliteal lymph node near the hilus. Mitotic figures were generally scarce. There was mild subcutaneous oedema of the ventral trunk extending from the axillae to the inner proximal thighs, which had not been evident clinically, and the lymph nodes (peripheral more so than internal) microscopically showed marked trabecular and perivascular fibrosis especially in hilar regions. Other congenital defects were hepatic capsular and central venous fibrosis with lymphatic duplication and dilatation in all areas of connective tissue, ventrally-incongruous half-circular tracheal rings, and multifocal renal dysplasia affecting the right kidney. There was locally-extensive subacute pyelonephritis of the left kidney.

Effects of long-term cyclosporin therapy on gingiva of rats: analysis by stereological and biochemical estimation

Cyclosporin A (CsA) is used as an immunosuppressive agent and its prominent side effect is the induction of gingival overgrowth, which remains a significant problem. The risk factors appraised include the duration of treatment. However, there are no stereological and biochemical studies exploring the effects of long-term CsA therapy on gingival tissue. The purpose of the present study was to investigate the level of TGF-beta1 in saliva and describe the densities of fibroblasts and collagen fibers in the gingival tissue of rats treated with CsA for long periods. Rats were treated for 60, 120, 180 and 240 days with a daily subcutaneous injection of 10 mg/kg of body weight of CsA. At the end of the experimental periods, saliva was collected for the determination of TGF-beta1 levels. After histological processing, the oral epithelium and the connective tissue area were measured as well as the volume densities of fibroblasts (Vf) and collagen fibers (Vcf). After 60 and 120 days of CsA treatment, there was a significant increase in Vf and Vcf as well as a significant increase in TGF-beta1 levels. After 180 and 240 days, reduction in the gingival overgrowth associated with significant decreases in the level of TGF-beta1, and also decreased Vf and Vcf, were observed. The data presented here suggest that after long-term therapy, a decrease in TGF-beta1 levels occurs, which might contribute to an increase in the proteolytic activity of fibroblasts in the gingiva, favoring the normality of extracellular matrix synthesis.

Fine Structure Histopathology of Labyrinthitis Ossificans in the Gerbil Model

Labyrinthitis ossificans (LO) is the pathological deposition of new bone within the lumen of the cochlea and labyrinth. This process occurs most commonly as a result of infection or inflammation affecting the otic capsule. Trauma and vascular compromise can also lead to neo-ossification within the otic capsule. The mechanism that regulates this process remains unestablished. This study details the end-stage histopathology in high-resolution plastic thin sections. Twenty Mongolian gerbils were infected by intrathecal injection of Streptococcus pneumoniae type 3 followed by subcutaneous penicillin G procaine (8 days) and were painlessly sacrificed 3 months later. The cochleas were serially divided and sectioned for light and electron microscopy. Sixteen of 20 animals (27 of 40 cochleas) demonstrated LO. Cochlear damage was most extensive in the vestibule and basal turn and decreased toward the apex, which often appeared normal. The histopathologic findings consisted of 1) new bone, calcospherites, osteoid, and fibrosis without dense connective tissue or osteoblasts extending from the endosteal wall into the lumen of the vestibule and scala tympani; 2) areas of dense connective tissue and osteoid enclosed by epithelial cells conjoined with the organ of Corti, stria vascularis, spiral ligament, and vestibular (Reissner's) membrane; and 3) partial to complete loss of the organ of Corti, spiral ligament cell bodies, stria vascularis, and spiral ganglion cells. Osteoblastic activity was not demonstrated in end-stage ossification in LO in the gerbil model. Neo-ossification appears to occur by calcospherite deposition along collagen-like fibrils within osteoid. The destruction of the organ of Corti, spiral ganglion cells, stria vascularis, and cells of Reissner's membrane and the spiral ligament occurs even in the absence of ossification of the cochlear duct.

O n-butil cianoacrilato na lobectomia pulmonar parcial em felinos: estudo experimental

O presente estudo teve como objetivo verificar a viabilidade do n-butil cianoacrilato na pneumostasia e hemostasia em lobectomia pulmonar parcial. Foram utilizadas seis gatas hígidas, adultas, com peso médio de três quilogramas. O procedimento cirúrgico consistiu na realização de toracotomia intercostal com lobectomia pulmonar parcial em que se utilizou adesivo cirúrgico para fechamento do lobo pulmonar. Após 21 dias da cirurgia, os animais foram novamente submetidos à toracotomia para a coleta de fragmento da região onde o adesivo fora aplicado, para análise histopatológica. Foram realizadas radiografias de tórax das pacientes para o acompanhamento pré e pós lobectomia pulmonar parcial e, aos dois, sete, 14 e 21 dias subseqüentes. As alterações radiográficas e clínicas encontradas como pneumotórax residual, aumento na densidade pulmonar, enfisema subcutâneo e apatia foram corrigidas em tempo hábil, não comprometendo os resultados do estudo. A histopatologia evidenciou reação inflamatória com predomínio de células mononucleares, neovascularização, proliferação de tecido conjuntivo e pequenas áreas de enfisema e atelectasia, sem comprometimento clínico significativo. Concluiu-se que o cianoacrilato é capaz de produzir adequada pneumostasia e hemostasia, e poderá ser uma opção para a realização de lobectomia pulmonar parcial em felinos.

Long-term denervation in humans causes degeneration of both contractile and excitation-contraction coupling apparatus, which is reversible by functional electrical stimulation (FES): a role for myofiber regeneration?

Over the last 30 years there has been considerable interest in the use of functional electrical stimulation (FES) to restore movement to the limbs of paralyzed patients. Spinal cord injury causes a rapid loss in both muscle mass and contractile force. The atrophy is especially severe when the injury involves lower motoneurons because many months after spinal cord injury, atrophy is complicated by fibrosis and fat substitution. In this study we describe the effects of long-term lower motoneuron denervation of human muscle and present the structural results of muscle trained using FES. By means of an antibody for embryonic myosin, we demonstrate that many regenerative events continue to spontaneously occur in human long-term denervated and degenerated muscle (DDM). In addition, using electron microscopy, we describe i) the overall structure of fibers and myofibrils in long-term denervated and degenerated muscle, including the effects of FES, and ii) the structure and localization of calcium release units, or triads; the structures reputed to activate muscle contraction during excitation-contraction coupling (ECC). Both apparatus undergo disarrangement and re-organization following long-term denervation and FES, respectively. The poor excitability of human long-term DDM fibers, which extends to the first periods of FES training, may be explained in terms of the spatial disorder of the ECC apparatus. Its disorganization and re-organization following long-term denervation and FES, respectively, may play a key role in the parallel disarrangement and re-organization of the myofibrils that characterize denervation and FES training. The present structural studies demonstrate that the protocol used during FES training is effective in reverting long-term denervation atrophy and dystrophy. The mean fiber diameter in FES biopsies is 42.2 +/- 14.8 SD (p < 0.0001 vs DDM 14.9 +/- 6.0 SD); the mean percentile of myofiber area of the biopsy is 94.3 +/- 5.7 SD (p < 0.0001 vs DDM 25.7 +/- 23.7 SD); the mean percentile fat area is 2.1 +/- 2.4 SD (p < 0.001 vs DDM 12.8 +/- 12.1 SD); and the mean percentile connective tissue area is 3.6 +/- 4.6 SD (p < 0.001 vs DDM 61.6 +/- 20.1 SD). In DDM biopsies more than 50% of myofibers have diameter smaller than 10 microm, while the FES-trained subjects have more that 50% of myofibers larger than 30 microm. The recovery of muscle mass seems to be the result of both a size increase of the surviving fibers and the regeneration of new myofibers.