Thick Connective Tissue Research Articles

Bladder augmentation with small intestinal submucosa leads to unsatisfactory long-term results

PurposeTo evaluate the use of small intestinal submucosa (SIS) for bladder augmentation in a series of select patients. Material and methodsSix patients (age 6.5–15.4, mean 9.8 years) underwent bladder augmentation with SIS: one after a cloacal exstrophy repair, one after multiple surgery of the bladder because of vesicoureteral reflux, two with spina bifida, two after bladder exstrophy repair. All suffered from a microbladder with a mean volume of 61.5 ml (range 15–120, 7–36% of expected bladder capacity for age). Preoperative bladder compliance ranged from 1.0 to 3.3 (mean 1.3) ml/cmH2O. ResultsFollow-up time ranged from 4.6 to 33.5 (mean 24.4) months. An increase of bladder volume was achieved in four patients (53–370 ml, 16–95% of expected bladder capacity for age). Bladder compliance postoperatively ranged from 0.9 to 5.6 (mean 3.0) ml/cmH2O. Histological examinations showed a complete conversion of SIS, leaving irregular urothelial lining and bladder wall containing muscular, vascular and relatively thick connective tissue in four patients and regular urothelium in two patients. Major complications were bladder stones in two patients and a bladder rupture in one patient. ConclusionBladder augmentation with SIS in humans failed to fulfill the hopes raised by animal studies. Due to the insufficient increase in bladder compliance and therefore failure to accomplish sufficient protection of the upper urinary tract, bladder augmentation with SIS cannot be recommended as a substitute for enterocystoplasty.

Limited Joint Mobility in Childhood Diabetes: Discovery, Description, and Decline

The discovery of limited joint mobility (LJM) as the earliest clinically apparent complication of diabetes in children and adolescents nearly 40 years ago provided insights into the mechanism of diabetes complications. LJM, due to periarticular connective tissue thickening and stiffness, varied from single joint involvement bilaterally to obvious hand deformity and limitation of movement of the spine. Thick, tight, waxy skin was apparent with more severe changes; biopsy indicated thickening of the dermis and epidermis with accumulation of collagen and loss of skin appendages. Substantial growth impairment was associated with all levels of LJM. Risk for microvascular disease over a 16-year period was actuarially increased nearly 4-fold by the presence of LJM. Long-term glycemic control influenced the onset of LJM; for every unit increase in average glycated hemoglobin from onset there was a 46% increase in the risk of LJM. Between 1976-78 and 1998, the prevalence of LJM decreased from 30 to 7%, and the portion with moderate to severe changes decreased from 33 to 14%, with the disappearance of severely affected individuals. Furthermore, growth data markedly improved for those with and without LJM; 30% without LJM and 77% with LJM had been less than the 25th percentile for height, whereas the more contemporary group had 22% without LJM and 33% with LJM in this quartile. The elucidation of LJM as a novel and informative early complication of pediatric diabetes has provided insights into the evolution of the severe long-term complications of diabetes. The marked decrease in the frequency of LJM and statural deficit in children and youth with diabetes in the 20 years between the late 1970s and 1990s can be attributed to improved metabolic control of children and adolescents. The prevalence of LJM in a population may serve as a measure of quality of diabetes control.

Acceleration of Wound Healing with Stem Cell–Derived Growth Factors

Recently, it has been revealed that bone marrow-derived mesenchymal stem cells (MSCs) accelerate the healing of skin wounds. Although the proliferative capacity of MSCs decreases with age, MSCs secrete many growth factors. The present study examined the effect of mesenchymal stem cell-conditioned medium (MSC-CM) on wound healing. The wound-healing process was observed macroscopically and histologically using an excisional wound-splinting mouse model, and the expression level of hyaluronic acid related to the wound healing process was observed to evaluate the wound-healing effects of MSC, MSC-CM, and control (phosphate-buffered saline). The MSC and MSC-CM treatments accelerated wound healing versus the control group. At 7 days after administration, epithelialization was accelerated, thick connective tissue had formed in the skin defect area, and the wound area was reduced in the MSC and MSC-CM groups versus the control group. At 14 days, infiltration of inflammatory cells was decreased versus 7 days, and the wounds were closed in the MSC and MSC-CM groups, while a portion of epithelium was observed in the control group. At 7 and 14 days, the MSC and MSC-CM groups expressed significantly higher levels of hyaluronic acid versus the control group (P < .05). The expression level of hyaluronic acid was lower at 14 days than at 7 days in all three groups. Both the MSC and MSC-CM groups accelerated wound healing versus the control group to a similar degree. Accordingly, it is suggested that the MSC-CM contains growth factor derived from stem cells, is able to accelerate wound healing as well as stem cell transplantation, and may become a new therapeutic method for wound healing in the future.

Distribution and fibrotic response following inhalation exposure to multi-walled carbon nanotubes.

BackgroundPrior studies have demonstrated a rapid and progressive acute phase response to bolus aspiration of multi-walled carbon nanotubes (MWCNTs). In this study we sought to test the hypothesis that inhalation exposure to MWCNT produces a fibrotic response and that the response is chronically persistent. To address the hypothesis that inhaled MWCNTs cause persistent morphologic changes, male C57BL/6 J mice were exposed in a whole-body inhalation system to a MWCNT aerosol and the fibrotic response in the alveolar region examined at up to 336 days after termination of exposure.MethodsInhalation exposure was to a 5 mg/m3 MWCNT aerosol for 5 hours/day for 12 days (4 times/week for 3 weeks). At the end of inhalation exposures, lungs were either lavaged for analysis of bronchoalveolar lavage (BAL) or preserved by vascular perfusion of fixative while inflated with air at 1, 14, 84, 168 and 336 days post inhalation exposure. Separate, clean-air control groups were also studied. Light microscopy, enhanced darkfield microscopy and field emission electron microscopy (FESEM) of tissue sections were used to analyze the distribution of lung burden following inhalation exposure. Morphometric measurements of Sirius Red staining for fibrillar collagen were used to assess the connective tissue response. Serial section analysis of enhanced darkfield microscope images was used to examine the redistribution of MWCNT fibers within the lungs during the post-exposure period.ResultsAt day 1 post-exposure 84 ± 3 and 16 ± 2 percent of the lung burden (Mean ± S.E., N = 5) were in the alveolar and airway regions, respectively. Initial distribution within the alveolar region was 56 ± 5, 7 ± 4 and 20 ± 3 percent of lung burden in alveolar macrophages, alveolar airspaces and alveolar tissue, respectively. Clearance reduced the alveolar macrophage burden of MWCNTs by 35 percent between 1 and 168 days post-exposure, while the content of MWCNTs in the alveolar tissue increased by 63 percent. Large MWCNT structures containing greater than 4 fibers were 53.6 percent of the initial lung burden and accounted for the majority of the decline with clearance, while lung burden of singlet MWCNT was essentially unchanged. The mean linear intercept of alveolar airspace, a measure of the expansion of the lungs, was not significantly different between groups. Pulmonary inflammation and damage, measured as the number of polymorphnuclear leukocytes (PMNs) or lactate dehydrogenase activity (LDH) and albumin in BAL, increased rapidly (1 day post) after inhalation of MWCNTs and declined slowly with time post-exposure. The fibrillar collagen in the alveolar region of MWCNT-exposed mice demonstrated a progressive increase in thickness over time (0.17 ± 0.02, 0.22 ± 0.02, 0.26 ± 0.03, 0.25 ± 0.02 and 0.29 ± 0.01 microns for 1, 14, 84, 168 and 336 days post-exposure) and was significantly different from clean-air controls (0.16 ± 0.02) at 84 and (0.15 ± 0.02) at 336 days post-exposure.ConclusionsDespite the relatively low fraction of the lung burden being delivered to the alveolar tissue, the average thickness of connective tissue in the alveolar region increased by 70% in the 336 days after inhalation exposure. These results demonstrate that inhaled MWCNTs deposit and are retained within the alveolar tissue where they produce a progressive and persistent fibrotic response up to 336 days post-exposure.

The worsening of tibialis anterior muscle atrophy during recovery post-immobilization correlates with enhanced connective tissue area, proteolysis, and apoptosis

Sustained muscle wasting due to immobilization leads to weakening and severe metabolic consequences. The mechanisms responsible for muscle recovery after immobilization are poorly defined. Muscle atrophy induced by immobilization worsened in the lengthened tibialis anterior (TA) muscle but not in the shortened gastrocnemius muscle. Here, we investigated some mechanisms responsible for this differential response. Adult rats were subjected to unilateral hindlimb casting for 8 days (I8). Casts were removed at I8, and animals were allowed to recover for 10 days (R1 to R10). The worsening of TA atrophy following immobilization occurred immediately after cast removal at R1 and was sustained until R10. This atrophy correlated with a decrease in type IIb myosin heavy chain (MyHC) isoform and an increase in type IIx, IIa, and I isoforms, with muscle connective tissue thickening, and with increased collagen (Col) I mRNA levels. Increased Col XII, Col IV, and Col XVIII mRNA levels during TA immobilization normalized at R6. Sustained enhanced peptidase activities of the proteasome and apoptosome activity contributed to the catabolic response during the studied recovery period. Finally, increased nuclear apoptosis prevailed only in the connective tissue compartment of the TA. Altogether, the worsening of the TA atrophy pending immediate reloading reflects a major remodeling of its fiber type properties and alterations in the structure/composition of the extracellular compartment that may influence its elasticity/stiffness. The data suggest that sustained enhanced ubiquitin-proteasome-dependent proteolysis and apoptosis are important for these adaptations and provide some rationale for explaining the atrophy of reloaded muscles pending immobilization in a lengthened position.

The possible protective effect of green tea versus interferon-α on carbon tetrachloride-induced hepatic fibrosis in albino rat

Introduction Liver fibrosis is a major health problem that can lead to liver cirrhosis, liver failure, and hepatocellular carcinoma. Aim of the work To study the possible protective effect of green tea extract versus interferon-α (IFN-α) in an animal model of carbon tetrachloride (CCl4)-induced hepatic fibrosis. Materials and methods The study was carried out on 60 adult male albino rats weighing 200–230 g. The rats were randomly divided into four main groups (15 rats each): group A included control rats, group B included rats that received an intraperitoneal injection of CCl4 (1 ml/kg body weight, twice weekly) to induce liver fibrosis, group C included rats that received both CCl4 (as in group B) and green tea extract (60 mg/rat/daily) by gavage, and group D included rats that received a subcutaneous injection of both CCl4 and IFN-α (100 000 IU/rat/daily). After 8 weeks, blood samples were collected for the assessment of serum levels of glutamic oxaloacetic transaminase (SGOT) and glutamic pyruvate transaminase (SGPT) and the liver was rapidly excised and processed for light and electron microscopic study. Results Light microscopic examination of group B liver sections showed that the liver parenchyma was disorganized and totally replaced by pseudolobules well demarcated by thick connective tissue septa. The majority of the hepatocytes were enlarged and showed a pale, markedly vacuolated cytoplasm and darkly stained small eccentric nuclei. The hepatocytes also showed loss of their PAS-positive glycogen granules. Electron microscopic study of the hepatocytes showed multiple large lipid globules compressing the nuclei and electron-dense mitochondria. In addition, Kupffer cells were increased in number. The intercellular space was widened and showed numerous Ito cells and abundant collagen fibers. In group C, the liver sections appeared more or less similar to those of the control group, whereas the hepatic tissue of group D showed intermediate histological alterations between both groups B and D. Biochemical data analysis supported the histological findings. Conclusion The current findings suggest that green tea exerts a more potent protective effect than IFN-α against CCl4-induced hepatic fibrosis. It may be recommended as an adjuvant therapy for patients with hepatic fibrosis.

The Role of Estrogen on Rat Parotid Gland after Menopause

Objective: To determine the histopathological changes of estrogen deficiency and estrogen replacement treatment in parotid gland of ovariectomized rats. Method: Female albino rats were divided into 3 groups: sham, ovariectomized, and ovariectomized with estrogen replacement. Histopathological evaluation of rat parotid glands were done by using the diameter of acinus and striated duct (SD), height of SD, and interacinar connective tissue thickness. Apoptosis in parotid gland were examined by TUNEL staining. Results: Except interacinar connective tissue thickness, there were no significant differences between the sham-operated control and ovariectomized group in terms of the diameter of acinus and striated duct (SD) and height of SD. Estrogen replacement resulted in significant increase in the height of SD and interacinar connective tissue thickness. Apoptotic index was higher in both ovariectomized and estrogen replacement group compared to sham-operated control. Conclusion: Although estrogen replacement did not reduce apoptotic index in parotid gland of overiectomized rat, it may have some positive influence on saliva secretion by increasing interaciner connective tissue thickness.

New dried human amniotic membrane is useful as a substitute for dural repair after skull base surgery.

Objectives Cerebrospinal fluid (CSF) leakage is an undesirable complication of skull base surgery. We used dried human amniotic membrane (AM) as a patch graft for dural repair to determine its efficacy in preventing CSF leakage. Design Frontoparietal craniotomy and removal of dura were performed in 20 Wistar rats. A dried AM was placed to cover the dural defect without suturing in 16 animals. In four animals, an expanded polytetrafluoroethylene was implanted. At 2 weeks and 1, 3, and 6 months, histological examination was performed. Dried AM was also used as a substitute in 10 patients who underwent skull base surgery, after approval by the Ethics Committee of the University of Toyama. Results At 2 weeks after implantation, thick connective tissue completely enclosed the dried AM. At 1 month after implantation, the connective tissue became thin and the implanted AM shortened. At 3 and 6 months after implantation, histological examination revealed disappearance of the dried AM and formation of membranous tissue. In the clinical study, neither CSF leakage nor clinical adverse reactions directly related to the dried AM were observed. Conclusion Dried human AM appears to be an ideal substitute for dura, since it is replaced by natural tissue.

Clinical and histological evaluation of subepithelial connective tissue after collagen sponge implantation in the human palate

Successful root-coverage treatment depends on the thickness of the donor tissue. This study aimed to evaluate the thickness of donor tissue after augmentation of the connective tissue in the palatal area by implantation of lyophilized collagen sponge (Hemospon(®) ). Ten patients with an indication for root coverage, whose palate was deficient in adequate connective tissue, were recruited. The procedure was carried out in two stages. In the first stage, the palatal thickness in the donor site was measured at three standardized points (points 1, 2 and 3), from the distal of the canine to the distal of the first molar, and the lyophilized collagen sponge was inserted. In the second stage, the palatal thickness over the implant was measured (at points 1, 2 and 3), two biopsies of the palatal mucosa were collected - one over the implant (experimental sample) and the other on the contralateral side (control sample) - and then root-coverage treatment was performed. Analyses consisted of clinical assessment of the palatal measurements before and after sponge implantation, and histological assessment of the experimental and control biopsy samples. Data were analyzed using the Wilcoxon test. Both analyses showed a significant increase in mean thickness, of 1.08 mm of neoformed tissue in the clinical analysis (the tissue at point 2 was the thickest of the three points) and of 0.53 mm in the histological analysis. The insertion of lyophilized collagen sponge induced a significant increase in the thickness of palatal connective tissue.

Effects of fibre type and kefir, wine lemon, and pineapple marinades on texture and sensory properties of wild boar and deer longissimus muscle

Fibre type percentage and changes in textural parameters, sensory properties as well as mean fibre cross sectional area (CSA), fibre shape, endomysium and perimysium thickness of wild boar and deer longissimus (L) muscle subjected to ageing with kefir, dry red wine, lemon and pineapple juice marinades for 4days were studied. Among the non-marinated and non-aged samples of muscles it was found that wild boar meat with its higher percentage of red fibres, higher CSA, thicker connective tissue as compared with deer meat, was harder, more springy and stringy.Muscles ageing, regardless of methods, resulted in a decrease in both the CSA and thickness of the connective tissue, and improve in fibre shape. As a consequence ageing caused a reduction in hardness, cohesiveness, springiness, and stringiness as well as in augmentation of tenderness, juiciness and general attractiveness of the muscles studied.As demonstrated by obtained data, regardless of ageing methods, deer L muscle contained more white fibres compared to wild boar muscle, were more susceptible to tenderization. The highest structural and textural changes, but the worst general attractiveness was found in muscles marinated with pineapple juice addition. Insignificantly lower changes in both quality traits were found in muscles aged with kefir marinade which at the same time were characterized by the high tenderness, the highest juiciness and general attractiveness.

Wyniki leczenia chirurgicznego nabytego zarośnięcia przewodu słuchowego zewnętrznego

Acquired post inflammatory external auditory canal atresia is a rear complication of external ear disease. It is presented as a connective tissue scar in bony part of external auditory canal. The aim of this study is clinical and epidemiological analysis and presentation of diagnostics and treatment results of patients with atresia treated in the Otolaryngology Department of Medical University of Gdansk. Material and methodsIn the period of 3 years (2008–11) 10 patients (8 women and 2 men) aged 30 to 70-years-old (medium 53 years) were treated for acquired atresia. All of these patients had medial closure of EAC with thick connective tissue scar and tympanic membrane involvement. Intrameatal access was performed in 5 patients, intraural access in 3 patients and retroaurical access in 2 patients. In all cases canaloplasty with meato-tympanic angle enlargement was performed, skin defect was replaced with free epidermal flap, silicon foil with fibrinous sponge was used for coverage. ResultsIn all of our patients external auditory canal widening and hearing improvement was achieved. Due to increasing EAC narrowing four patients underwent second surgery with another free epidermal flap grafting. ConclusionThe selection of surgical access in patients with acquired atresia should comply with the etiology of the disease and the shape of EAC. During surgery we aim at maximal broadening of the bony part of EAC. The success of the treatment depends on meato-tympanic angle enlargement and free epidermal flap grafting.

Factoring-in agglomeration of carbon nanotubes and nanofibers for better prediction of their toxicity versus asbestos

BackgroundCarbon nanotubes (CNT) and carbon nanofibers (CNF) are allotropes of carbon featuring fibrous morphology. The dimensions and high aspect ratio of CNT and CNF have prompted the comparison with naturally occurring asbestos fibers which are known to be extremely pathogenic. While the toxicity and hazardous outcomes elicited by airborne exposure to single-walled CNT or asbestos have been widely reported, very limited data are currently available describing adverse effects of respirable CNF.ResultsHere, we assessed pulmonary inflammation, fibrosis, oxidative stress markers and systemic immune responses to respirable CNF in comparison to single-walled CNT (SWCNT) and asbestos. Pulmonary inflammatory and fibrogenic responses to CNF, SWCNT and asbestos varied depending upon the agglomeration state of the particles/fibers. Foci of granulomatous lesions and collagen deposition were associated with dense particle-like SWCNT agglomerates, while no granuloma formation was found following exposure to fiber-like CNF or asbestos. The average thickness of the alveolar connective tissue - a marker of interstitial fibrosis - was increased 28 days post SWCNT, CNF or asbestos exposure. Exposure to SWCNT, CNF or asbestos resulted in oxidative stress evidenced by accumulations of 4-HNE and carbonylated proteins in the lung tissues. Additionally, local inflammatory and fibrogenic responses were accompanied by modified systemic immunity, as documented by decreased proliferation of splenic T cells ex vivo on day 28 post exposure. The accuracies of assessments of effective surface area for asbestos, SWCNT and CNF (based on geometrical analysis of their agglomeration) versus estimates of mass dose and number of particles were compared as predictors of toxicological outcomes.ConclusionsWe provide evidence that effective surface area along with mass dose rather than specific surface area or particle number are significantly correlated with toxicological responses to carbonaceous fibrous nanoparticles. Therefore, they could be useful dose metrics for risk assessment and management.

An autopsy study to clarify characteristics of local recurrence after extended pancreatectomy with intraoperative radiation therapy in patients with pancreatic cancer

Local relapses frequently occur even after curative resection of pancreatic cancer. To control local recurrence, we adopted extended radical resection combined with intraoperative radiation therapy. A retrospective review was conducted on 41 patients who underwent extended radical pancreatectomy combined with intraoperative radiation therapy for pancreatic cancer. Fourteen patients underwent autopsies. We took en bloc specimens of the abdominal aorta with surrounding connective tissue to evaluate histological characteristics of local status at autopsies. Autopsies disclosed microscopic local recurrence in five (36%) of the 14 patients, although no evidence of local relapse was observed in either follow-up images or macroscopic findings at autopsy. Of the three patients with R1 resection, two had no local recurrence microscopically at autopsy. Histological features of local recurrence in autopsy samples showed small numbers of cancer cells surrounded by thick connective tissue without mass formation. The autopsy study revealed that a characteristic of local recurrence after this treatment was tiny cancer cells scattered in dense connective tissue; these cells were undetected by follow-up imaging.

Expressions of bax, bcl-2 and ki-67 in odontogenic keratocysts (keratocystic odontogenic tumor) in comparison with ameloblastomas and radicular cysts

The aim of the study was to determine the apoptotic features and proliferation potential of odontogenic keratocysts compared with ameloblastomas and radicular cysts by analysing the role of bax, bcl-2, and Ki-67. The study material consisted of 20 odontogenic keratocysts, 20 radicular cysts, and 20 ameloblastomas. Immunohistochemically, bax, bcl-2 and Ki-67 were applied. The positive cells were evaluated in both neoplastic/nonneoplastic odontogenic epithelium and connective tissue cells. Ameloblastoma showed stronger bcl-2 expression than odontogenic keratocysts and radicular cysts. Bcl-2 expression in the whole thickness of epithelium and connective tissue of odontogenic keratocyst was significantly higher than radicular cyst. The expression of bax in the epithelium of radicular cyst was significantly higher than odontogenic keratocyst and ameloblastoma. The lining epithelium of odontogenic keratocyst showed stronger Ki-67 expression than ameloblastoma and radicular cyst. The proliferation potential of the epithelium and the overexpression of various anti-apoptotic proteins in odontogenic epithelial tumors are quite significant for their clinical behaviour. High expressions of bcl-2 and Ki-67 in odontogenic keratocysts accord with their aggressive clinical behaviour and a high recurrence rate.

Sonographic Measurements of Subsynovial Connective Tissue Thickness in Patients With Carpal Tunnel Syndrome

A major pathologic finding in patients with idiopathic carpal tunnel syndrome is noninflammatory fibrosis and thickening of the subsynovial connective tissue. The objective of this study was to determine the ability of sonography to depict this thickening by comparing subsynovial connective tissue thickness in patients with carpal tunnel syndrome and healthy control participants. Longitudinal sonograms of the middle finger superficial flexor tendon and subsynovial connective tissue were obtained at 3 levels: at the wrist crease (proximal tunnel), at the hook of the hamate (mid tunnel), and at the distal edge of the transverse carpal ligament (distal tunnel). The thickness of the subsynovial connective tissue perpendicular to the direction of the tendon and the diameter of the flexor digitorum superficialis tendon at the same level were measured. Then, a thickness ratio was created. At all 3 levels, the subsynovial connective tissue was thicker in patients than in controls (P < .0001) with a thickness ranging from 0.60 to 0.63 mm in patients and 0.46 to 0.50 mm in controls. The thickness ratio was significantly greater in patients at the hamate and distal levels (P = .018 and .013, respectively). With this study, we have shown that it is possible to measure subsynovial connective tissue thickness with sonography, and the tissue is thicker in patients with carpal tunnel syndrome than in healthy controls.

132 A NEW APPROACH TO ASSESS OVARIAN STRUCTURES IN LIVE RABBITS: SURGICAL TRANSLOCATION AND ULTRASOUND BIOMICROSCOPY

For the purpose of determining factors associated with ovulation induction in rabbits, there is a need to examine the ovaries in a serial fashion. To this end, a study was designed to develop an approach that would permit serial evaluation of ovarian structures in vivo using ultrasound biomicroscopy (UBM) while preserving ovarian function in rabbits. The use of UBM has enabled characterisation of ovarian dynamics in live mice and has recently been validated for assessing structures in excised rabbit ovaries (Cervantes et al. 2011 Reprod. Fertil. Dev. 23, 183–184). However, acoustic impedance of the body wall prevented acquisition of high-quality ovarian images in vivo by transabdominal UBM in rabbits. Female New Zealand White rabbits (5 months old; n = 12) underwent surgical ovarian translocation. Each ovary was exteriorized through a lateral flank incision on the respective sides, without compromising vascular supply. The mesovarium at each pole of the ovary was transfixed to the muscle layers of the abdominal wall to keep the ovary in a subcutaneous position. Finally, the subcutaneous and skin layers were closed. No postoperative complications were observed. The surgical site was allowed to heal for 2 weeks and the skin sutures were removed before UBM was attempted. The rabbits were wrapped in a towel for restraint without sedation. Acoustic gel was applied to the shaved skin over the translocated ovary and ovarian structures were imaged transcutaneously by UBM using a 25-MHz transducer on 3 consecutive days. Both ovaries were clearly distinguished in all 12 rabbits during each examination. Antral follicles ranging from 0.6 to 3.1 mm were detected and the number of follicles ≥0.6 mm per pair of ovaries ranged from 7 to 18. Data were analysed by one-way ANOVA. The number and diameter of follicles ≥0.6 mm per rabbit did not differ among the 3 examinations (mean ± s.e.m.; 11.0 ± 0.82, 11.8 ± 0.85 and 12.3 ± 0.75 follicles and 1.3 ± 0.07, 1.3 ± 0.07 and 1.4 ± 0.07 mm, respectively). However, follicles ≥2 mm in diameter were detected in 3 out of 12 rabbits on the first and second days and in 5 out of 12 rabbits on the third day. One or more corpora lutea were detected in the same 2 rabbits (2/12) during each of the 3 examination days. Two months after surgery, ovarian structures were clearly discernible by UBM in 11 of 24 translocated ovaries. Image deterioration was attributed to the attenuation of sound waves associated with increased skin thickness and growth of subcutaneous fat and connective tissue around the ovary, assessed at necropsy. In summary, translocated ovaries continued to function in their new location and transcutaneous UBM permitted serial visualisation of ovarian structures in live rabbits. Therefore, this new approach provides a nonterminal alternative for repetitive examination of rabbit ovaries and is particularly appropriate for experiments involving factors associated with ovulation induction in rabbits. This research was supported by the Natural Sciences and Engineering Research Council of Canada.

Injection of Acanthaster planci with thiosulfate-citrate-bile-sucrose agar (TCBS). II. Histopathological changes

We assessed histological changes in the tissues of the crown-of-thorns starfish Acanthaster planci (COTS) after injection of thiosulfate-citrate-bile-sucrose agar (TCBS) which was used as a disease inducer (potential outbreak control method), by conventional and scanning electron microscopy. Digestive glands were processed and stained with hematoxylin and eosin to describe the histological architecture of the intestinal epithelium. Subsequently comparison of healthy versus infected tissues and Gram stains were carried out to confirm bacterial occurrence on infected tissues, characterize the structural changes induced by bacterial communities in COTS tissues, and to determine if the histopathological changes of intestinal tissues were consistent with vibrio infection. TCBS injections induced marked epithelial desquamation, hypertrophy and hypersecretion of glandular cells, epithelial cell destruction, pyknosis, reduction of thickness and disorganization of connective tissue and associated nerve plexus, presence of bacterial colonies, irregular eosinophilic foci in glandular cells, brush border disruption, atrophy and detachment of intestinal microvilli and cell debris in the lumen. All these changes were attributed to a fulminating systemic dysbiosis and were consistent with vibrio infections.

Reduced thoracolumbar fascia shear strain in human chronic low back pain

BackgroundThe role played by the thoracolumbar fascia in chronic low back pain (LBP) is poorly understood. The thoracolumbar fascia is composed of dense connective tissue layers separated by layers of loose connective tissue that normally allow the dense layers to glide past one another during trunk motion. The goal of this study was to quantify shear plane motion within the thoracolumbar fascia using ultrasound elasticity imaging in human subjects with and without chronic low back pain (LBP).MethodsWe tested 121 human subjects, 50 without LBP and 71 with LBP of greater than 12 months duration. In each subject, an ultrasound cine-recording was acquired on the right and left sides of the back during passive trunk flexion using a motorized articulated table with the hinge point of the table at L4-5 and the ultrasound probe located longitudinally 2 cm lateral to the midline at the level of the L2-3 interspace. Tissue displacement within the thoracolumbar fascia was calculated using cross correlation techniques and shear strain was derived from this displacement data. Additional measures included standard range of motion and physical performance evaluations as well as ultrasound measurement of perimuscular connective tissue thickness and echogenicity.ResultsThoracolumbar fascia shear strain was reduced in the LBP group compared with the No-LBP group (56.4% ± 3.1% vs. 70.2% ± 3.6% respectively, p < .01). There was no evidence that this difference was sex-specific (group by sex interaction p = .09), although overall, males had significantly lower shear strain than females (p = .02). Significant correlations were found in male subjects between thoracolumbar fascia shear strain and the following variables: perimuscular connective tissue thickness (r = -0.45, p <.001), echogenicity (r = -0.28, p < .05), trunk flexion range of motion (r = 0.36, p < .01), trunk extension range of motion (r = 0.41, p < .01), repeated forward bend task duration (r = -0.54, p < .0001) and repeated sit-to-stand task duration (r = -0.45, p < .001).ConclusionThoracolumbar fascia shear strain was ~20% lower in human subjects with chronic low back pain. This reduction of shear plane motion may be due to abnormal trunk movement patterns and/or intrinsic connective tissue pathology. There appears to be some sex-related differences in thoracolumbar fascia shear strain that may also play a role in altered connective tissue function.

Histologic changes in the sinus membrane after maxillary sinus augmentation with simultaneous implant placement using engineered bone graft material

The aim of the study was to analyze the histologic and ultrastructural changes after maxillary sinus augmentation with simultaneous implant placement using engineered bone graft material. In this study, calcium phosphate cement (CPC) scaffolds combined with goat bone marrow stromal cells (BMSCs) were used to fill goat sinus floor space after maxillary sinus floor elevation with simultaneous implant placement comparing with those not filled any grafted materials and used as controls. After a healing period of 3 months, the goat maxillary sinus membrane was examined using light microscopy and scanning electronic microscopy. The results showed that the connective tissue thickness and the epithelium thickness of mucosa were not statistically significant difference between two groups. The tissue engineered bone complex might be an ideal graft for the sinus floor elevation and have no influence on the sinus membrane under the histological and ultrastructural observation.

Spontaneous Thymoma in a Juvenile Cynomolgus Macaque (Macaca fascicularis)

A single, solid, yellow-white thymic mass was found at necropsy of a two-year-old female cynomolgus macaque from a four-week, repeat-dose toxicity and immunogenicity study. Microscopically, the mass was multilobular and well encapsulated, surrounded by a thick connective tissue capsule, and composed of dense sheets of elongate or spindle-shaped cells and large cystic cavities separated by thick connective tissue stroma. Normal thymus was adjacent to the mass, but it was compressed. Within the mass were abundant interspersed Hassall's corpuscles; individual and small clusters of mature, small lymphocytes; scattered eosinophils; large areas of necrosis; focal mineralization; and cholesterol clefts. An interesting feature was the presence of large multinucleated giant cells, which varied widely in size and nuclear number. Immunohistochemical staining for two lymphocyte markers and two structural proteins confirmed the identity of the neoplastic spindle cells and other cellular components. There was no evidence of vascular invasion or metastasis. Features of the thymoma indicated it was a pre-existing condition and not treatment related.