Outer Connective Tissue Research Articles

Degeneration of muscle spindles in a murine model of Pompe disease

Pompe disease is a debilitating medical condition caused by a functional deficiency of lysosomal acid alpha-glucosidase (GAA). In addition to muscle weakness, people living with Pompe disease experience motor coordination deficits including an instable gait and posture. We reasoned that an impaired muscle spindle function might contribute to these deficiencies and therefore analyzed proprioception as well as muscle spindle structure and function in 4- and 8-month-old Gaa−/− mice. Gait analyses showed a reduced inter-limb and inter-paw coordination in Gaa−/− mice. Electrophysiological analyses of single-unit muscle spindle proprioceptive afferents revealed an impaired sensitivity of the dynamic and static component of the stretch response. Finally, a progressive degeneration of the sensory neuron and of the intrafusal fibers was detectable in Gaa−/− mice. We observed an increased abundance and size of lysosomes, a fragmentation of the inner and outer connective tissue capsule and a buildup of autophagic vacuoles in muscle spindles from 8-month-old Gaa−/− mice, indicating lysosomal defects and an impaired autophagocytosis. These results demonstrate a structural and functional degeneration of muscle spindles and an altered motor coordination in Gaa−/− mice. Similar changes could contribute to the impaired motor coordination in patients living with Pompe disease.

Bionic artificial penile Tunica albuginea

Bionic artificial penile Tunica albuginea

Neuroimmune cardiovascular interfaces control atherosclerosis.

Atherosclerotic plaques develop in the inner intimal layer of arteries and can cause heart attacks and strokes1. As plaques lack innervation, the effects of neuronal control on atherosclerosis remain unclear. However, the immune system responds to plaques by forming leukocyte infiltrates in the outer connective tissue coat of arteries(the adventitia)2-6. Here, because the peripheral nervous system uses the adventitia as its principal conduit to reach distant targets7-9, we postulated that the peripheral nervous system may directly interact with diseased arteries. Unexpectedly, widespread neuroimmunecardiovascular interfaces (NICIs) arose in mouse and human atherosclerosis-diseased adventitia segments showed expanded axon networks, including growth cones at axon endings near immune cells and media smooth muscle cells. Mouse NICIs established a structural artery-brain circuit (ABC): abdominal adventitia nociceptive afferents10-14 entered the central nervous system through spinal cord T6-T13 dorsal root ganglia and were traced to higher brain regions, including the parabrachial and central amygdala neurons; and sympathetic efferent neurons projected from medullary and hypothalamic neurons to the adventitia through spinal intermediolateral neurons and both coeliac and sympathetic chain ganglia. Moreover, ABC peripheral nervous system components were activated: splenic sympathetic and coeliac vagus nerve activities increased inparallel to disease progression, whereas coeliac ganglionectomy led to the disintegration of adventitial NICIs, reduced disease progression and enhanced plaque stability. Thus, the peripheral nervous system uses NICIs to assemble a structural ABC, and therapeutic intervention in the ABC attenuates atherosclerosis.

Pathology, bacteriology and molecular studies on caseous lymphadenitis in Camelus dromedarius in the Emirate of Abu Dhabi, UAE, 2015-2020.

Caseous lymphadenitis (CLA) or pseudotuberculosis is a chronic zoonotic bacterial disease caused by Corynebacterium pseudotuberculosis, which affects livestock and humans. This study aimed to describe the pathology, bacteriology and confirm the identity of the pathogen by 16S rRNA gene sequencing in Camelus dromedarius. A total of 12 camels with suspected CLA in three regions of Abu Dhabi Emirate (Abu Dhabi, Al Ain and Al Dhafra), United Arab Emirate (UAE) were subjected to clinical and postmortem examinations from January 2015 to December 2020. Clinically, camels were emaciated and showed the presence of external caseous abscesses suggestive of CLA. Postmortem examination showed multiple abscesses of variable sizes with caseous material encapsulated by fibrous tissue in the liver, lungs, muscle, and lymph nodes. Following clinical and postmortem examination, blood, pus and different tissue samples were collected for subsequent analysis. Histopathological examination of all organs stained with Hematoxylin and Eosin (H&E) indicated a central caseo-necrotic core that was admixed with bacterial colonies and infiltration of chronic inflammatory cells, surrounded by a pyogenic membrane, and an outer fibrous connective tissue capsule. Bacterial culture identified the isolates of Corynebacterium pseudotuberculosis biotype ovis strain, and these isolates were shown to be sensitive to all antibiotics tested (penicillin, ampicillin, Co-trimoxazole, enrofloxacin and tetracycline). Moreover, the identity of the isolates was confirmed by partial sequencing of the 16S rRNA gene which showed a 100% identity to Corynebacterium pseudotuberculosis. Phylogenetic analysis based on 16S rRNA gene sequence clearly differentiates Corynebacterium pseudotuberculosis from other species of Corynebacterium. Briefly, this study provided the basic information for infection of Corynebacterium pseudotuberculosis in Camels and will help in controlling of this pathogen in the region.

First Report of Peripheral Nerves in Bone from Triceratops horridus Occipital Condyle

Peripheral nerves in vertebrate systems have been studied for centuries. Nerve anatomy, including the connective tissues that cushion nerves, are well described in the literature. The outer connective tissues, known as the epineurium and perineurium, include a double layer of collagen fibers that form a double helical wrapping that is visible with polarized light microscopy and by other contrast methods. This wrapping is a diagnostic characteristic of nerve tissues. Decades of research into dinosaur bones have produced interesting endocasts of probable brain morphology, but no soft and flexible nerve tissues have been reported from dinosaur remains. In this study we provide evidence of nerve fragments, characterized by a double helical wrapping of collagen fibers, from a Triceratops condyle collected at the Hell Creek Formation, MT. Based on comparison with nerves from an avian model we conclude that these are fragments of nerves that once resided in vivo in Triceratops.

Influence of Mesenchymal Stem Cells Seeding on Morphology of Tissue-Engineered Vascular Graft Based on Poly (L-lactide) Scaffold

Introduction - The problem of small-diameter vascular grafts still exists in modern vascular surgery. Autologous or synthetic vascular grafts are used routinely for arterial bypass in patients with cardiovascular diseases. However, some patients either lack suitable autologous tissue or cannot receive synthetic grafts. Such patients could benefit from a vascular graft produced by tissue engineering. The aim of the study is to evaluate the effects of adipose-derived mesenchymal stem cells (AD-MSC) on the formation of tissue-engineered vascular graft (TEVG). Methods - Tubular scaffolds 1.1 mm inner diameter were obtained by the electrospinning method on the basis of nano- and microfibers of poly(L-lactide) (PLLA). AD-MSC were seeded on the scaffolds by the developed filtration method. The subsequent culturing for 14 days was carried out in the constructed flow bioreactor. Cell distribution in the scaffold wall was assessed by the fluorescence microscopy. Obtained grafts were implanted into rat aorta: group 1 without AD-MSC (n=36) with follow-up till 16 months, group 2 with seeded AD-MSC (n=36) with follow-up till 16 months. The material was subjected to histological examination, electron microscopy, immunohistochemistry CD 31+, aSMA and morphometric analysis (cell counting, neointima and neoadventia thickness). To track AD-MSC in vivo PKH-26 labeling and subsequent fluorescence microscopy were performed in 2, 7 and 14 days. Results - Filtration seeding and subsequent cultivation in the flow bioreactor led to a uniform distribution of AD-MSC in the scaffold wall. Graft patency in the first group was 86%, in the second 96%. All grafts formed neointima, with no signs of hyperplasia in the area of anastomoses. The total resorption of PLLA fibers in group 1 was observed by histological examination, new vascular wall consisted of endothelial lining and connective tissue without smooth muscle cells (SMC); in all cases aneurysm formation was detected. Total cells number of implant wall was more in group 2, including SMC that formed neomedia. Also the outer connective tissue layer was much thicker. Histology identified the uniform formation of new tissues in group 2 with no signs of aneurysm. Conclusion - AD-MSC seeding leads to the formation of TEVG, morphologically similar to the natural structure of the vessels. The study was supported by Russian Science Foundation (project 14-33-00003)

Morphology, Morphometry and Histogenesis of the Prenatal Dromedary (Camelus Dromedarius) Spleen

Abstract Prenatal gross morphologic, morphometric and histologic developmental features of the dromedary spleen were studied. The dromedary gestation period (13 months) was categorized into four (1-4) phases and ten developing spleens per growth phase were sampled. Splenic topographical anatomy was noted before being eviscerated from each foetus. Morphologic and morphometric features of the eviscerated spleens were immediately documented and 2 – 4 mm thick samples were collected for histological analysis. The developing spleen was dark brown in colour, semilunar shaped and significantly increased (p<0.05) in size and weight across the four phases of prenatal development. The full-term dromedary spleen was observed to have unique histological features. Its capsule had an inner smooth muscle and an outer predominant connective tissue layer. The pumping of stored blood from the muscular capsule and trabeculae was proportionate to the body’s requirement. The splenic venous return was characterized by blood flow from the red pulp (venous sinusoids) to the peritrabecular sinuses, subcapsular sinuses and finally to the splenic veins. The dromedary has a sinusal type of spleen and has both open and closed types of circulation. The presence of closed circulation and absence of marginal sinus could be the reason for dromedary main health problems of blood parasites; Trypanosoma evansi. It was concluded that most of the salient features of the postnatal spleen were already evident in the first growth phase and became developed by the second phase. Other growth phases were mainly characterized by increase in sizes.

Localization of the ultraviolet-sensor Opn5m and its effect on myopia-related gene expression in the late-embryonic chick eye

Recent studies show that exposure to ultraviolet (UV) light suppresses ocular elongation, which causes myopia development. However, the specific mechanisms of this process have not been elucidated. A UV-sensor, Opsin 5 (Opn5) mRNA was shown to be present in extraretinal tissues. To test the possibility that UV-signals mediated by Opn5 would have a direct effect on the outer connective tissues of the eye, we first examined the expression patterns of a mammalian type Opn5 (Opn5m) in the late-embryonic chicken eye. Quantitative PCR showed Opn5m mRNA expression in the cornea and sclera. The anti-Opn5m antibody stained a small subset of cells in the corneal stroma and fibrous sclera. We next assessed the effect of UV-A (375 nm) irradiation on the chicken fibroblast cell line DF-1 overexpressing chicken Opn5m. UV-A irradiation for 30 min significantly increased the expression of Early growth response 1 (Egr1), known as an immediate early responsive gene, and of Matrix metalloproteinase 2 (Mmp2) in the presence of retinal chromophore 11-cis-retinal. In contrast, expression of Transforming growth factor beta 2 and Tissue inhibitor of metalloproteinase 2 was not significantly altered. These results indicate that UV-A absorption by Opn5m can upregulate the expression levels of Egr1 and Mmp2 in non-neuronal, fibroblasts. Taken together with the presence of Opn5m in the cornea and sclera, it is suggested that UV-A signaling mediated by Opn5 in the extraretinal ocular tissues could influence directly the outer connective tissues of the chicken late-embryonic eye.

Functional ultrastructure of the parenchymatic capsules of the cestode Thysanotaenia congolensis (Cyclophyllidea, Anoplocephalidae, Inermicapsiferinae).

Proglottids from adult Thysanotaenia congolensis from naturally infected black rats Rattus rattus from Santiago Island, Cape Verde, were examined by transmission electron microscopy (TEM). The uterus in mature proglottids is composed of an ephemeral transverse tube or sac that breaks down, releasing eggs into the parenchyma where they are sequestered in groups and encapsulated by parenchymatous layers. In gravid proglottids, eggs accumulate in groups of 6-12 at the distal end of sac-like uterine ducts. As eggs accumulate, the end of the uterine ducts expands until it pinches off, releasing groups of eggs into the parenchyma surrounded by remnants of uterine epithelium. These epithelium-bound groups of eggs remain in the parenchyma until they are encapsulated with several parenchymatous layers, forming parenchymatic egg capsules, typical for mature and gravid proglottids of Inermicapsiferinae. The parenchymatic capsules originate from the medullary parenchyma of immature proglottids, which undergoes differentiation into the three layers of gravid proglottids: (1) an outer connective tissue layer composed of long delicate filaments of unknown chemical nature embedded in a granular extracellular matrix; (2) a middle layer appearing as an accumulation of large closely packed PAS-positive mucous goblets that are intensely metachromatic after toluidine blue staining and (3) an inner compact layer composed of lipid-containing cells, muscle cells with elongated muscle fibres and cells of various sizes and shapes forming or containing calcareous corpuscles. The mature hexacanths of T. congolensis are surrounded by reduced oncospheral envelopes consisting of remnants of a very thin membranous layer of degenerating embryophore with long, irregularly shaped cytoplasmic processes and by remnants of uterine epithelium extending as numerous apical microlamellae into the parenchymatic capsule lumen.

High‐Resolution Median Nerve Sonographic Measurements

To study relationships between median wrist and forearm sonographic measurements and median nerve conduction studies. The study population consisted of a prospective convenience sample of healthy adults. Interventions included high-resolution median nerve sonography and median motor and sensory nerve conduction studies. Main outcome measures included median motor nerve compound muscle action potential amplitude, distal latency, and conduction velocity; sensory nerve action potential amplitude and distal latency; and sonographic median nerve cross-sectional area. Median motor nerve and sensory nerve conduction studies of the index finger were performed using standard published techniques. A second examiner blinded to nerve conduction study results used a high-frequency linear array transducer to measure the cross-sectional area of the median nerve at the distal volar wrist crease (carpal tunnel inlet) and forearm (4 cm proximally), measured in the transverse plane on static sonograms. The outer margin of the median nerve was traced at the junction of the hypoechoic fascicles and adjacent outer connective tissue layer. Fifty median nerves were evaluated in 25 participants. The compound muscle action potential amplitude with wrist stimulation was positively related to the cross-sectional area, with the area increasing by 0.195 mm(2) for every millivolt increase in amplitude in the dominant hand (95% confidence interval, 0.020, 0.370 mm(2); P < .05) and 0.247 mm(2) in the nondominant hand (95% confidence interval, 0.035, 0.459 mm(2); P < .05). There was no significant linear association between the wrist median cross-sectional area and median motor and sensory distal latencies. Conduction velocity through the forearm was not significantly linearly associated with the forearm area or forearm-to-wrist area ratio (tapering ratio). The wrist area was inversely related to the sensory nerve action potential amplitude. Although associations were found between median nerve conduction study amplitudes and sonographic nerve measurements, they were not found for other parameters. Studying these relationships may increase our understanding of when to best use these procedures.

Hox11 genes are required for regional patterning and integration of muscle, tendon and bone

Development of the musculoskeletal system requires precise integration of muscles, tendons and bones. The molecular mechanisms involved in the differentiation of each of these tissues have been the focus of significant research; however, much less is known about how these tissues are integrated into a functional unit appropriate for each body position and role. Previous reports have demonstrated crucial roles for Hox genes in patterning the axial and limb skeleton. Loss of Hox11 paralogous gene function results in dramatic malformation of limb zeugopod skeletal elements, the radius/ulna and tibia/fibula, as well as transformation of the sacral region to a lumbar phenotype. Utilizing a Hoxa11eGFP knock-in allele, we show that Hox11 genes are expressed in the connective tissue fibroblasts of the outer perichondrium, tendons and muscle connective tissue of the zeugopod region throughout all stages of development. Hox11 genes are not expressed in differentiated cartilage or bone, or in vascular or muscle cells in these regions. Loss of Hox11 genes disrupts regional muscle and tendon patterning of the limb in addition to affecting skeletal patterning. The tendon and muscle defects in Hox11 mutants are independent of skeletal patterning events as disruption of tendon and muscle patterning is observed in Hox11 compound mutants that do not have a skeletal phenotype. Thus, Hox genes are not simply regulators of skeletal morphology as previously thought, but are key factors that regulate regional patterning and integration of the musculoskeletal system.

Transurethral partial cystectomy with a 2 micron laser in diagnosis and treatment for bladder submucosal lesions in adults

To investigate the characteristics of transurethral partial cystectomy with a 2 µm laser in diagnosis and treatment for the bladder submucosal lesions in adults. Nine patients with suspected pathological diagnosed bladder submucosal lesions in out-patient department were diagnosed and treated transurethral with a 2 µm laser under sacral block between August 2009 and December 2010. The diameters of tumors were 1.5 - 2.5 cm. A 2 µm laser was used to incise the full-thickness bladder wall around the tumors. The entire bladder wall was peeled between the detrusor muscle layer and outer connective tissues. Tumors with bladder wall at the base were removed together and sent for pathological examination. The surgical procedures, intraoperative hemorrhage, intraoperative and postoperative complications were observed, pathological diagnosis and postoperative follow-up were performed. All operations were successful. Mean operative time was 36.4 minutes (range 25 to 47 minutes), perioperative blood loss was minimal. There was no obturator nerve reflection and no hemorrhage detected after surgery. Postoperative pathological diagnosis included leiomyoma in 3 cases, pheochromocytoma in 3 cases, endometriosis in 1 case and metastatic bladder cancer in 2 cases. Transurethral partial cystectomy with a 2 µm laser can diagnose and treat bladder submucosal lesions. The procedures are effective and safe. Patients could get accurate pathological diagnosis without further painful and some bladder tumors can be treated by minimally invasive surgery.

Transurethral Partial Cystectomy with 2 μm Thulium Continuous Wave Laser in the Treatment of Bladder Pheochromocytoma

To introduce transurethal partial cystectomy with a 2013 nm thulium laser as a treatment for paraganglioma of the urinary bladder in adults. Three patients with pheochromocytomas were treated transurethrally with a 2013 nm thulium laser under general anesthesia. A 2013 nm thulium laser was used to incise the full-thickness bladder wall around the tumors. The entire bladder wall was peeled between the detrusor muscle layer and outer connective tissues. Tumors with full-thickness detrusor muscle layers at the base were removed together. Intraoperative fluctuation of blood pressure, preoperative values of 24-hour urine catecholamine (CA) and vanillylmandelic acid (VMA), and postoperative complications were observed, and postoperative followtwoups were performed. All operations were successful. Operative time was 25 to 32 minutes. Perioperative blood pressure was stable in two cases while blood pressure fluctuated in the third case. When the entire tumor and the full-thickness bladder wall at the base were freed, blood pressure reverted to stability. All values of 24-hour urine CA and VMA were within normal limits postoperatively. Patients were followed for 7 to 9 months postoperatively with no recurrence. This series included highly selected patients who were treated by a single senior surgeon who is rich in experience in performing 2013 nm thulium laser procedures. To our knowledge, this is the first report of a 2013 nm thulium laser used to treat bladder pheochromocytoma. It can be applied to precisely vaporize and incise the full-thickness bladder wall and cut down the blood supply of the tumor, then peel it while blood pressure remains stable, thus completing partial cystectomy for bladder pheochromocytoma safely.

1555 THE RISK OF IATROGENIC POSITIVE MARGIN IN RADICAL PROSTATECTOMY: AN EXPENDED CRITERION TO EVALUATE THE QUALITY OF SURGICAL RESECTION

You have accessJournal of UrologyProstate Cancer: Localized VI1 Apr 20101555 THE RISK OF IATROGENIC POSITIVE MARGIN IN RADICAL PROSTATECTOMY: AN EXPENDED CRITERION TO EVALUATE THE QUALITY OF SURGICAL RESECTION Christian Barré, Geneviève Aillet, and Matthieu Thoulouzan Christian BarréChristian Barré Nantes, France More articles by this author , Geneviève AilletGeneviève Aillet Nantes, France More articles by this author , and Matthieu ThoulouzanMatthieu Thoulouzan Toulouse, France More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2010.02.1320AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Conventionally, the rate of positive surgical margins (PSM) for pT2 tumors, i.e. iatrogenic positive margins (IPM), is taken as the technical error rate. However, this is not a good indicator of technical error as it concerns dissection errors involving only malignant and not benign tissue. The technical error rate should also include the risk of IPM i.e. all incisions of benign tissue within the histological limits of the prostate defined by McNeal because the incised tissue might just as well have been malignant as benign. Even if the benign proved to be benign, this did not eliminate the risk that was run. The safety of a technique does not depend only on the known consequences of an error such as a PSM (″adverse events″) but also on the number of times errors with potentially deleterious consequences are made (″near-misses″). The aim of this prospective study was to establish expanded criteria for evaluating the quality of surgical resection in patients undergoing radical retropubic prostatectomy (RRP). METHODS Prospective study (Jan 2005-Dec 2007), 507 consecutive patients with localised prostate cancer underwent RRP. We determined the rate of IPM and PSM. Pathological study: whole-mount 3-mm serial sections using the Stanford technique. Where capsule was absent (apex and bladder neck) no malignant or benign gland must be in contact with the inked margin. In the capsular zone, we defined 1st degree capsule incision (1st DCI): loss of the outer connective tissue layer and 2nd degree capsule incision (2nd DCI): loss of both capsule layers with malignant or benign gland in contact with ink. RESULTS Patient characteristics were mean age 61.3 years; disease stage: pT2 73%, pT3 25.8%; bilateral nerve-sparing n=273, 88.1%, unilateral n=37, 11.9%. The risk of IPM was 5%: incision of benign glandular tissue was 2.8% (apex: 43%; bladder neck: 57%), 1st DCI was 2.2% and 2nd DCI was 0%. The overall PSM rate was 6.3%: 2.2 % (8/370) for pT2, 14.5 % (19/131) for pT3, and 83.3% for pT4. It was 4.8% (15/310) in patients undergoing nerve-sparing surgery. PSM with pT2 tumours occurred at apex (1.7%), postero-lateral 0%, bladder neck (0.5%). PSMs with pT3a/b tumours occurred at: apex (6.1%), postero-lateral (3.8%) and bladder neck (6.9%). PSM size distribution for pT2: PSM≤1 mm: n=4; 1<PSM≤4 mm: n=2; PSM >4 mm: n=2, and for pT3: PSM≤1 mm: n=6; 1<PSM≤4 mm: n=5; PSM>4 mm: n=8. Biochemical recurrence rate (PSA >0.10 ng/ml) at 1 year was 2.6%. CONCLUSIONS The safety of an RP procedure is given not only by the PSM rate for pT2 tumours but also by the percentage of times there is a risk of a PSM. © 2010 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 183Issue 4SApril 2010Page: e600 Advertisement Copyright & Permissions© 2010 by American Urological Association Education and Research, Inc.MetricsAuthor Information Christian Barré Nantes, France More articles by this author Geneviève Aillet Nantes, France More articles by this author Matthieu Thoulouzan Toulouse, France More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Transurethral Partial Cystectomy With Continuous Wave Laser for Bladder Carcinoma

To our knowledge we established a new technique to vaporize and incise the full-thickness bladder wall and peel off the entire muscular layers with a 2 microm continuous wave laser to treat bladder carcinoma. Nine patients were treated transurethrally with a 2 microm laser under sacral block. This 2 microm laser was used to incise the full-thickness bladder wall around the tumors. The entire bladder wall was peeled between the detrusor muscle layer and outer connective tissues. Tumors with full-thickness detrusor muscle layers at the base were removed together and sent for pathological examination. Intraoperative hemorrhage and postoperative complications were observed, and pathological staging and postoperative followup were performed. All operations were successful. Mean +/- SD operative time was 8.7 +/- 2.6 minutes (range 7 to 15) per patient. Perioperative blood loss was minimal. There was no obturator nerve reflection and no hemorrhage was detected after surgery. Bladder perforation occurred in 1 case. Pathological stage of the tumors was T1 and T2a. Patients were followed for 6 to 9 months postoperatively with no recurrence at the resection site. This series included highly selected patients treated by a single high volume surgeon. To our knowledge this is the first report of a 2 microm laser used to treat bladder carcinoma. It can be applied to precisely vaporize and incise the full-thickness bladder wall and peel it between detrusor muscle layers and outer loose connective tissues, thus completing partial cystectomy for bladder carcinoma.

Molecular characterization and analysis of a truncated serotonin receptor gene expressed in neural and reproductive tissues of abalone

In molluscs, the neurotransmitter serotonin (5-HT) has been linked to a variety of biological roles including gamete maturation and spawning. The possible involvement of 5-HT in abalone gamete release was demonstrated by a dose-dependent increase in Haliotis rubra gonad contractile bioactivity following 5-HT stimulation. Physiological functions associated with 5-HT, are mediated through binding to 5-HT receptors. A cDNA encoding a putative 5-HT receptor consisting of 359 amino acids was isolated from the tropical abalone H. asinina, termed 5-HT(1 ha). The 5-HT(1 ha) shares G-protein-coupled receptor motifs with metazoan 5-HT receptors, including predicted transmembrane domains, active sites for protein kinase action, and N-linked glycosylation sites. However, the third intracellular loop of 5-HT(1 ha) is relatively short, and only six transmembrane domains are predicted, implying a truncated receptor. Phylogenetic analysis with known 5-HT receptor genes suggests that 5-HT(1 ha) belongs to the type 1 5-HT receptor family. Expression analysis by RT-PCR showed that 5-HT(1 ha) mRNA was present in all tissues examined, including the neural ganglia and gonad tissues. Immunocytochemistry revealed the presence of 5-HT(1 ha) specifically within the soma of neuronal cells located in the outer cortex of both cerebral and pleuropedal ganglia. In ovarian and testicular tissues, 5-HT(1 ha) immunoreactivity was observed in epithelial cells of the outer capsule and connective tissue of the trabeculae to which the gamete follicles adhere. Whether this receptor transcript is translated to a functional protein needs to be verified, but if so, it could play a role in reproduction.

Activation of Na,K,2Cl‐cotransporter stimulates outer medullary connective tissue expression

Nitric oxide (NO) deficiency stimulates renal medullary Na,K,2Cl‐cotransporter (NKCC2) activity. Chronic NO synthase inhibition has also been shown to increase the production of extracellular matrix components, thus promoting renal fibrosis. The role of NKCC2 in this process is not fully understood. We here hypothesize that chronic activation of NKCC2 leads to increased production of extracellular matrix components despite intact NO production. To test this, male adult Brattleboro rats with central diabetes insipidus were treated with the vasopressin 2 receptor analoque desmopressin (5ng/h; 5d), via osmotic minipumps. Urinary output served as a measure of treatment efficiency. NKCC2 expression in outer medullary tissue samples was determined by TaqMan real time RT PCR. Microarray analysis was used to identify differentially regulated genes. In the treated rats 24h urine output was decreased to 6% (p=.01) and outer medullary NKCC2 expression was increased to 170% (p=.013) of controls. Microarray analysis revealed increased expression of numerous extracellular matrix components including collagen I, III–VI, XII, XV and XVIII (&gt;2fold; p&lt;10‐5). We conclude that the activation of NKCC2 in a setting of intact local NO synthesis is associated with increased outer medullary connective tissue production. Long term studies are needed to elucidate the pathophysiological significance of this finding.

The morphology of the penis in porcupine (Hystrix cristata)

: In this study, histological and macroanatomical structures of porcupine penis were examined on 3&amp;nbsp;adult male porcupines. The non-erectile position of porcupine penis was directed to caudal. There was an obvious collum penis. Glans penis with a black colour was easily distinguished from corpus penis which had a white color. There were cornified papillae covering on both the glans penis and corpus penis. Two small nail symmetrical structures were noted in connective tissue just behind of glans penis, and under the urethra of the corpus penis were distinguished. Histological examinations indicated that these structures histologically fulfilled completely to the name as the nail contained an inner and an outer layer, namely, connective tissue and keratin, respectively. Histological evaluations also showed that it is suitable to categorize the porcupines penis as vascular type. In conclusion, the anatomy of porcupine penis was initially examined in the present study. Our findings will make an important contribution to the knowledge of similar subjects.

Biodegradable polydioxanone and poly( l/ d)lactide implants: an experimental study on peri-implant tissue response

Several implants for orbital wall fracture treatment are available at the present, but they have drawbacks: resorption, risk for migration and foreign body reaction. Alloplastic resorbable implants would be advantageous: no removal operation and no donor side morbidity. The purpose of this study was to evaluate the foreign body reaction, capsule formation and mechanical properties of two bioresorbable implants. PDS and SR-P(L/DL)LA mesh sheet (70/30) with solid frame (96/4) implants (SR-P(L/DL)LA 70,96) were placed into subcutaneous tissue of 24 rats. Immunohistochemistry was used to evaluate reactivity for Tn-C, α-actin, type I and III collagens and two mononuclear cells: T-cells and monocyte/ macrophage. GPC, DSC and SEM were performed. Student's t-test or nonparametric Kruskall–Wallis test were used for statistical analysis. Histology of peri-implant capsule exhibited an inner cell-rich zone and an outer connective tissue zone around both materials. Tn-C reactivity was high in the inner and α-actin in the outer zone. At the end of the study, the difference of type I collagen versus type III collagen reactivity in inner zone was statistically significant ( P < 0.0001) as was the difference of type I collagen versus type III collagen reactivity in outer zone ( P < 0.0001). Immunohistochemistry did not reveal any statistical differences of T-cell and monocyte/macrophage reactivity around PDS versus SR-P(L/DL)LA 70,96 implants, nor any differences as a function of time. PDS were deformed totally after 2 months. SR-P(L/DL)LA 70,96 implants were only slightly deformed during the follow up of 7 months. PDS degraded rapidly in SEM observation. Particles were detaching from surface. SEM observation revealed that polylactide implant was degrading from the surface and the inner porous core became visible. The degradation came visible at 7 months. There were cracks in perpendicular direction towards to the long axis of the filaments. M w of PDS decreased fast compared to the polylactide implant. Foreign body reaction was minimal to both materials but continued throughout the whole observation period. Mechanically PDS was poor, it looses its shape totally within 2 months. It cannot be recommended for orbital wall reconstruction. New mesh sheet–frame structure (SR-P(L/DL)LA 70,96) approved to be mechanically adequate for orbital wall reconstruction. It seems not to possess intrinsic memory and retains its shape. The resorption time is significantly longer compared to PDS and is comparable to other studied P(L/DL)LA copolymers. Thus, the new polylactide copolymer implant may support the orbital contents long enough to give way to bone growth over the wall defect.

Histologic features of capsule in breast implant

目的 通过动物实验,以了解硅凝胶乳房假体包膜中胶原组织学特点.方法 30只大白兔,每只大白兔背部两侧肌层下随机各置入10 ml硅凝胶假体1个.3周、1、2、3、6、12个月时分别将包膜和假体完整地取出,在光学显微镜下测量包膜厚度,取包膜样本做成石蜡切片,进行HE染色、James染色、维多利亚蓝、丽春红复合染色和天狼猩红染色,光学显微镜下观察比较包膜特点,其余包膜样本进行胶原分型和含量测定.结果⑴假体组织学特点:大致可分为两层,包膜外层为疏松结缔组织,内层为致密结缔组织.包膜厚度在3个月前逐渐增加(P<0.01);置入时间越长,假体包膜胶原成分越多、细胞成分越少,包膜越厚(P<0.01);3个月后,趋于稳定(P＞0.05). ⑵包膜胶原总量在3个月前逐渐增高(P<0.01);3个月后趋于稳定(P＞0.05).⑶3个月前,包膜中胶原纤维最多,网状纤维次之,弹力纤维最少甚至无;, 3个月后趋于稳定.⑷3个月前,包膜Ⅲ型胶原有减少趋势,Ⅰ型胶原有增多趋势;3个月后,均趋于稳定.结论假体包膜在组织学上与瘢痕类似,是假体周围创面愈合的必然产物。