A novel mouse model that recapitulates adult-onset glycogenosis type 4.

Glycogen storage disease type IV (GSD IV) is a rare autosomal recessive disorder caused by deficiency of the glycogen-branching enzyme (GBE). The diagnostic hallmark of the disease is the accumulation of a poorly branched form of glycogen known as polyglucosan (PG). The disease is clinically heterogeneous, with variable tissue involvement and age at onset. Complete loss of enzyme activity is lethal in utero or in infancy and affects primarily the muscle and the liver. However, residual enzyme activity as low as 5-20% leads to juvenile or adult onset of a disorder that primarily affects the central and peripheral nervous system and muscles and in the latter is termed adult polyglucosan body disease (APBD). Here, we describe a mouse model of GSD IV that reflects this spectrum of disease. Homologous recombination was used to knock in the most common GBE1 mutation p.Y329S c.986A > C found in APBD patients of Ashkenazi Jewish decent. Mice homozygous for this allele (Gbe1(ys/ys)) exhibit a phenotype similar to APBD, with widespread accumulation of PG. Adult mice exhibit progressive neuromuscular dysfunction and die prematurely. While the onset of symptoms is limited to adult mice, PG accumulates in tissues of newborn mice but is initially absent from the cerebral cortex and heart muscle. Thus, PG is well tolerated in most tissues, but the eventual accumulation in neurons and their axons causes neuropathy that leads to hind limb spasticity and premature death. This mouse model mimics the pathology and pathophysiologic features of human adult-onset branching enzyme deficiency.

Distinctive immunostaining of claudin-4 in spiradenomas.

The intercellular bridges are essential structures in maintaining the histologic organization of the epithelium, while providing a very efficient way to exchange molecules between cells and transduction of the cell-to-cell and matrix-to-cell signals. Derangement in those important structures' physical integrity and/or function, which can be assessed by the presence or absence of several intercellular bridge proteins including claudin-4, E-cadherin, and ß-catenin, was found to be related to several phenomena in the path to the neoplastic transformation. However, these proteins have not been studied in the wide variety of the skin neoplasms, in detail. Herein, we immunohistochemically assessed the expression patterns of these 3 intercellular bridge proteins on a total of 86 epidermal and eccrine adnexal tumors including basal cell carcinoma, squamous cell carcinoma, poroma, spiradenoma, syringoma, and hidradenoma. We observed a selective and distinct claudin-4 expression in the ductal-type cells of all cases of spiradenomas. Similarly, in the poromas, syringomas, and hidradenomas, claudin-4 was only positive in the luminal cells of microcystic structures, although not as conspicuous as in the spiradenomas. On the other hand, E-cadherin and ß-catenin were positive in almost all types of the tumors, in a way which was not contributory to differentiate from each other. In conclusion, we think that claudin-4 can be helpful at least in making a reliable differential diagnosis of spiradenoma when overlapping morphologic features do not allow to further subclassification in the overwhelming variety of the adnexal tumors.

NFU1 gene mutation and mitochondrial disorders.

Mitochondrial respiratory chains consist of approximately 100 structural proteins. Thirteen of these structural proteins are encoded by mitochondrial DNA (mtDNA), and the others by nuclear DNA (nDNA). Mutation in any of the mitochondrial structural-protein related genes, regardless of whether they are in the nDNA or mtDNA, might cause mitochondrial disorders. In the recent past, new nuclear genes required for assembly, maintenance, and translation of respiratory chain proteins have been found. Mutation in these genes might also cause mitochondrial disorders (MD). NFU1 gene is one of such genes and has a role in the assembly of iron-sulfur cluster (ISC). ISCs are included in a variety of metalloproteins, such as the ferredoxins, as well as in enzymatic reactions and have been first identified in the oxidation-reduction reactions of mitochondrial electron transport. It is important to be aware of NFU1 gene mutations that may cause severe mitochondrial respiratory chain defects, mitochondrial encephalomyopathies and death, early in life.

Effect of VEGF gene therapy and hyaluronic acid film sheath on peripheral nerve regeneration.

Functional nerve regeneration after reconstructive nerve surgery remains unsatisfying. In this study, vascular endothelial growth factor (VEGF) gene therapy combined with a hyaluronic acid (HA)-enriched microenvironment in nerve regeneration was investigated. Sciatic nerve was transected, and end-to-end neurorrhaphy was performed on 32 male Sprague-Dawley rats, which were randomly divided into four groups (n = 8 per group): nerve coaptation without treatment (group I); nerve coaptation covered with HA film sheath (group II); nerve coaptation with intramuscular VEGF gene in plasmid injection (group III); and nerve coaptation combined with HA film sheath and intramuscular VEGF gene in plasmid injection (group IV). Contralateral sciatic nerves were used as control. VEGF expression was verified from gluteal muscle biopsies surrounding the sciatic nerve by reverse transcriptase-PCR. Electrophysiological, histopathological, and electron microscopic evaluations were performed after 4 weeks. Mean peak amplitude of groups I-IV and nonoperated sciatic nerve were 4.5 ± 0.6 mV, 6.4 ± 0.4 mV, 6.7 ± 0.5 mV, 8.5 ± 0.4 mV, and 9.8 ± 0.5 mV, respectively. Mean myelinated axonal counts of groups I-IV and nonoperated sciatic nerve were 105 ± 24, 165 ± 19, 181 ± 22, 271 ± 23, and 344 ± 17, respectively. Treatment with HA film sheath coverage combined with intramuscular VEGF gene in plasmid injection yielded statistically significant higher peak amplitudes and myelinated axonal counts (P < 0.001). In addition, significantly less scar formation with HA administration (groups II and IV; P < 0.001) was found. Thus, it was found that VEGF might crucially regulate nerve regeneration processes and that HA can reduce the scar formation. This study showed that the combination of HA film sheath and VEGF gene may synergistically promote peripheral nerve regeneration.

Hourglass-like constrictions of the radial nerve in the neuralgic amyotrophy: A case report.

Neuralgic amyotrophy (NA) is a peripheral nerve disorder that has a classical presentation as motor deficit after severe pain, but it is still overlooked or misdiagnosed. Formerly, the diagnosis was based on the clinical picture and electrophysiology; however, sophisticated imaging and surgical modalities showed structural abnormalities such as hourglass-like constrictions of the nerves. In this article, we present a case presenting with drop hand mimicking radial nerve entrapment. The patient was diagnosed with NA and surgery revealed hourglass-like constrictions. The clinical findings were improved after neurorrhaphy and physical therapy. In conclusion, hourglass-like constrictions can be prognostic factors of NA and should be searched carefully.

Bilateral renal metastasis of an inguinal malignant solitary fibrous tumor, 9 years after primary surgical treatment.

OBJECTIVE: To report the first case of a bilateral renal solitary fibrous tumor (SFT) as a metastasis of an inguinal malignant SFT. CLINICAL PRESENTATION AND INTERVENTION: A 60-year-old male patient with a history of a right inguinal 7 × 8 cm soft tissue mass excision 9 years ago was referred to our clinic with abdominal pain. Both physical examination and chest X-ray were normal. Computed tomography revealed bilateral renal tumor. He was successfully treated with left partial and right radical nephrectomy. Histopathological examination showed a metastasis of the previous inguinal SFT. CONCLUSION: This case showed that although malignant SFT is extremely rare in the urogenital tract, this tumor should be included in the differential diagnosis when identified in the kidneys.

Efficacy of melatonin, mercaptoethylguanidine and 1400W in doxorubicin- and trastuzumab-induced cardiotoxicity.

Doxorubicin (DOX) and Trastuzumab (TRAST) are effective agents for the treatment of many neoplastic diseases. Cardiotoxicity is a major side effect of these drugs and limit their use. In this study, the possible protective effects of melatonin (MEL), mercaptoethylguanidine (MEG), or N-(3-(aminomethyl) benzyl) acetamidine (1400W) against the cardiotoxicity of DOX and TRAST were tested. Male Sprague-Dawley rats received an injection of DOX (20 mg/kg) alone or in combination with TRAST (10 mg/kg) to induce cardiotoxicity; daily treatments with MEL (10 mg/kg × 2), MEG (10 mg/kg × 2), or 1400W (10 mg/kg × 2) were begun 36 hr before and continued for 72 hr after DOX and TRAST administration. Oxidant/antioxidant indices of the cardiac tissue, namely, malondialdehyde, superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), as well as serum levels of creatine phosphokinase (CK-MB) were measured. Additionally, the injury scores were evaluated histopathologically. Malondialdehyde levels were significantly higher, while SOD and GSH-Px activities were significantly reduced in rats with DOX- or DOX+TRAST-induced cardiotoxicity compared to normal values. All three treatment agents significantly reversed oxidative stress markers. Serum CK-MB levels were significantly increased after treatment with DOX and DOX+TRAST; these changes were also reversed by each of the treatments and resulted in near normal levels. Both the DOX- and DOX+TRAST-treated rats presented similar histopathologic injuries; in the animals treated with the protective agents, histologic protection of the cardiac tissue was apparent. These results suggested that MEL, MEG, as well as 1400 W are effective in preventing DOX- or DOX+TRAST-induced cardiotoxicity.

Comparison of hyperbaric oxygen and medical ozone therapies in a rat model of experimental distal colitis.

OBJECTIVES: Previous studies have shown that hyperbaric oxygen (HBO) is effective in reducing the severity of acute distal colitis (ADC). Ozone therapy (OT) reduces inflammation in several pathological conditions. We aimed to compare the effects of HBO therapy and OT in an experimental ADC rat model. MATERIALS AND METHODS: Forty rats were randomly divided into four groups: Sham, ADC, ADC + HBO, and ADC + OT. Rats in the sham group were given isotonic saline. In the remaining groups, ADC was created by intracolonic administration of 4% acetic acid. No treatment was given to the ADC group. The rats in the ADC + HBO and ADC + OT groups were given HBO and ozone treatments, respectively. The administration of acetic acid caused an inflammatory response in all animals. Distal colons and blood samples were obtained. RESULTS: The histopathological score was significantly higher in the ADC group compared to the other groups. The histopathological scores in the ADC + HBO and ADC + OT groups were significantly lower compared to the ADC group (both p < 0.001). The most pronounced therapeutic effect was observed in the ADC + OT group. Malondialdehyde and neopterin levels and superoxide dismutase and glutathione peroxidase activities in the ADC group were significantly higher compared to the other groups (p < 0.001). CONCLUSION: Our data showed that the therapeutic effect of OT is more pronounced than that of HBO therapy. Its possible effect is by means of decreasing inflammation, edema, and oxidative stress. These findings also suggest that it is possible to improve the outcome of ADC by using ozone therapy as an adjuvant therapy.

Diagnostic importance of MRI and CT scans for synovial osteochondromatosis of the temporomandibular joint.

Since synovial chondromatosis (SC) clinically mimics symptoms of internal derangements of the TMJ, the diagnostic value of MRI and CT, overlooked for years, is discussed in the presented case. Multiple amorphous calcifications in the left infratemporal fossa and upper synovial compartment of the TMJ were detected on the CT and MRI scans. The patient underwent open TMJ arthrotomy and removal of 15 calcified loose bodies. SC may be diagnosed radiographically when sclerosis of the glenoid fossa, soft tissue edema, and intraarticular radio-opaque loose bodies are detected. Advanced imaging of the TMJ, such as MRIs and CTs, are indispensible methods to obtain differential diagnoses for long-standing suspicious pathologies of the temporomandibular joint.

Melatonin ameliorates doxorubicin-induced skin necrosis in rats.

Doxorubicin (DXR), a highly effective chemotherapeutic agent, causes serious injury when extravasated. The injury can sometimes result in skin necrosis and ulceration, requiring surgery. The detrimental effect of DXR on the antioxidant system via free oxygen radicals is one of the mechanisms proposed in its etiology. Thus, we used melatonin, a potent antioxidant, and compared the effects with dimethylsulfoxide (DMSO), which is used in the treatment of patients with DXR-induced extravasation.Twenty-seven Wistar-albino rats were used. After intradermal injection of DXR, DMSO was injected into the extravasated area and melatonin was given intraperitoneally. On day 14 of the experiment, skin ulcers were clearly formed and samples were taken with a punch biopsy. Ulcer sizes were measured. Tissue samples were analyzed for superoxide dismutase, glutathione peroxidase, and malondialdehyde enzymes, and histopathologically evaluated.Melatonin clearly decreased MDA levels, ulcer size, and histopathologic ulcer scores in DXR extravasated tissue. DMSO also decreased MDA levels, ulcer size and histopathologic ulcer score. However, melatonin was remarkably more effective than DMSO in terms of antioxidant enzyme activity, oxidative stress, and histopathologic ulcer scores in rats. Necrosis was evident in the DXR-treated group and some slides showed necrosis involving the fascia. Histopathologic ulcer scores of the necrotic tissue decreased in the DMSO and melatonin groups. The ulcer score in the melatonin group was significantly lower than in the control group. Although the ulcer score in the DMSO group was lower than control, there was no statistically significant difference. The ulcer size in the DMSO group was significantly lower than the control group. The ulcer size in the melatonin group was significantly lower than both the DMSO and control groups.We believe that melatonin, either alone or in combination with DMSO, may be used for treating DXR extravasation. In addition, free oxygen radicals play a crucial role in the etiology of the injury, which should be considered in further studies.

Comparisons of the effects of systemic administration of L-thyroxine and doxycycline on orthodontically induced root resorption in rats.

The aim of this study was to histologically evaluate and compare the effects of the systemic administration of L-thyroxine (TX) and doxycycline (DX) on orthodontically induced root resorption, Twenty-eight male 50- to 60-day-old Wistar rats were used. Seven rats served as the baseline control. Seven animals received TX (20 µg/kg bodyweight/day) and seven DX (1.2 mg/kg bodyweight/day), by means of a mini-osmotic pump implanted subcutaneously. Seven rats were separated as a sham, in order to evaluate the pure effect of the surgical procedure on the animals' health. Tooth movement (TM) was achieved with a continuous force of 50 g by placing Elgiloy coil springs between the right maxillary first molar and incisors for 14 days. The animals were sacrificed and specimens containing the appliance and maxillary tooth-bearing segments were processed for light microscopy. The surface area of root resorption lacunae was measured histomorphometrically using digital photomicrographs. To evaluate the resorptive changes on the molar root surface of each group, scanning electron microscopy (SEM) examinations were also carried out. Statistical evaluation of root resorption percentages was performed using Kruskal-Wallis analysis of variance test. Multiple comparisons were determined by the Student-Newman-Keuls method. The level of significance was set at P < 0.05. Histomorphometric analysis of root resorption, expressed as a percentage, showed that the average relative root resorption affecting the maxillary molars on the TM side was 0.32 ± 0.25 in the TX and 0.26 ± 0.06 in the DX groups and 2.19 ± 0.86 in the control. Statistically significant inhibition of root resorption was determined both in the TX and DX groups (P < 0.001) on the TM side. There was no statistically significant difference in relative root resorption between the TX and DX groups. Systemic administration of TX and DX demonstrated similar effects on root resorption in rats and may have inhibitory effects on orthodontically induced resorptive activity.

Effects of hyperbaric oxygen therapy on rabbit skeletal muscle during extremity lengthening.

INTRODUCTION: Extremity lengthening through distraction osteogenesis is limited by the surrounding skeletal muscle and neurovascular structures rather than the bone itself. The purpose of this study is to evaluate the effects of hyperbaric oxygen therapy on skeletal muscle during distraction osteogenesis. MATERIALS AND METHODS: Twenty New Zealand white rabbits were randomly divided into two groups. Right tibia of all rabbits was distracted at a rate of 0.125 mm per 6 h (0.5 mm/day) for 10 days with circular external fixator. Experimental group rabbits (N=10) underwent 2.5 ATA hyperbaric oxygen therapy for 2 h everyday for 20 days, control group rabbits (N=10) did not receive any corresponding treatment. Skeletal muscle perfusion was evaluated with scintigraphy before and after the distraction period. Serum CPK, LDH and AST levels were measured before and after the distraction period. All animals were killed on the 27th day. The right tibias of all animals were removed and tibialis posterior muscle was harvested for histopathologic and histomorphometric assessment with light and electron microscopy. RESULTS: Skeletal muscle perfusion was decreased in the control group in comparison with pre-distraction level (P=0.008). However, no significant decrease was observed in the experimental group (P=0.678). There were no statistical differences in serum CPK, LDH and AST levels between groups (P=0.340, P=0.077, P=0.796). The mean area of the muscle fibers was measured as 398.66+/-9.16 micro2 in the experimental group and 349.44+/-5.76 micro2 in the control group (P=0.000) with light microscopy. Mild fibrosis was observed in connective tissue component of muscle tissue in control group. An average of 26 myofibrils (20-32) was counted in a 16-cm2 unit area in experimental group and 50 myofibrils (35-65) in the control group with electron microscopy. Enlargement in the sarcoplasmic reticulum, degenerative changes in nuclear cytoplasm and increase in myofibril diameter were observed in the control group, which was not observed in the experimental group CONCLUSION: Results of this study suggest that HBO treatment alleviates the detrimental effects of distraction on skeletal muscles and preserves its ultrastructure.

Anatomical assessment of bile ducts of Luschka in human fetuses.

Bile ducts of Luschka (also called subvesical or supravesicular ducts) can cause bile leakage during laparoscopic cholecystectomy, especially if surgery is carried out in ignorance of such variations. The aim of this study was to clarify the clinical anatomy of these ducts in human fetuses and frequency of the ducts locating near gallbladder fossa. Thirty-two fetal cadaver livers were dissected and the gallbladders were separated from the livers and ducts were investigated under a surgical microscope. All observed ducts were examined microscopically and connective tissue cords were excluded. Bile ducts of Luschka locating near cystic fossa were found in 7 of 32 fetuses (21.9%). Three of the seven ducts ran towards to liver segment 5 (S5); three ducts were found in the gallbladder fossa; and one duct ran towards to liver segment 4 (S4). Also it was found that three of the seven ducts drained into the subsegmental duct of S5, two ducts drained into the right hepatic duct, one duct drained into the right anterior branch bile duct, and one duct drained into the subsegmental duct of S4. Subvesical ducts running along the gallbladder fossa between the gallbladder and the liver parenchyma were found in a relatively high incidence in fetuses than adults. Awareness and knowledge about incidence of such ducts alerts the surgeon during laparoscopic cholecystectomy. Therefore morbidity due to bile leaks can be reduced.

Does the type of resuscitative fluid affect healing of colonic anastomosis in experimentally induced hemorrhagic shock in rats?

OBJECTIVE: The aim of the study was to investigate the effects of different resuscitative fluids on the healing of intestinal anastomosis in a hemorrhagic-shock rat model. MATERIALS AND METHODS: Closed-colony Wistar male rats (n = 40; 8 rats per group) were subjected to volume-controlled hemorrhagic shock, followed by a 30-min shock phase. The animals were then resuscitated with one of the following fluids (which also corresponds to their respective groups): lactated Ringer's solution (LR), hydroxyethyl starch (HES), 7.5% hypertonic saline (HS) and autologous blood (AB). There was also a control group (CL), which did not experience hemorrhagic shock or receive any resuscitative fluids. All rats underwent laparotomy, segmental resection and anastomosis of the left colon. Five days later, a 2nd laparotomy was performed and the anastomotic bursting pressure was measured in vivo. Thereafter, the anastomosed segment was resected to measure the tissue hydroxyproline level and the grade of anastomotic fibrosis. RESULTS: All experimental groups (LR, HES, HS and AB) exhibited lower anastomotic bursting pressures than the CL group; however, no intergroup differences achieved statistical significance. The mean tissue hydroxyproline level and fibrosis grade also were similar across all 5 groups. CONCLUSION: In traumatic hemorrhagic shock, anastomosis safety does not appear to be affected by the type of fluid used for resuscitation. Moreover, LR, HES and HS all seemed to reinforce healing as effectively as transfused blood.

Choanal polyp originating from the nasal septum: septochoanal polyp.

Most of the nasal polyps arise from the lateral walls of the nasal cavity. Nasal polyps originating from the nasal septum with choanal extension are extremely rare. We report a case of large choanal polyp that arised from the posterosuperior aspect of the nasal septum, and extended down to the oropharynx. A 52-year-old woman presented with a two-year history of progressive nasal obstruction and snoring. Findings of anterior rhinoscopy were in normal limits. We think that the term "septochoanal polyp" which, as far as we know, has not been mentioned in the literature before, can be used for this rare lesion.

Demonstration of technical feasibility and viability of whole eye transplantation in a rodent model.

INTRODUCTION: Whole eye transplantation (WET) holds promise for vision restoration in devastating/disabling visual loss (congenital or traumatic) not amenable to surgical or neuroprosthetic treatment options. The eye includes multiple tissues with distinct embryonic lineage and differential antigenicity. Anatomically and immunologically, the eye is unique due to its avascular (cornea) and highly vascular (retina) components. Our goal was to establish technical feasibility, demonstrate graft viability, and evaluate histologic changes in ocular tissues/adnexae in a novel experimental model of WET that included globe, adnexal, optic nerve (ON), and periorbital soft tissues. METHODS: Outbred Sprague-Dawley rats (n = 5) received heterotopic vascularized WET from donors. Each WET included the entire globe, adnexa, ON, and periorbital soft tissues supplied by the common carotid artery and external jugular vein. Viability and perfusion were confirmed by clinical examination, angiography and magnetic resonance imaging (MRI). Globe, adnexal, and periorbital tissues were analyzed for histopathologic changes, and the ON was examined for neuro-regeneration at study endpoint (30 days) or Banff Grade 3 rejection in the periorbital skin (whichever was earlier). RESULTS: Gross examination confirmed transplant viability and corneal transparency. Average operative duration was 64.0 ±â€¯5.8 min. Average ischemia time was 26.0 ±â€¯4.2 min. MRI revealed loss of globe volume by 36.0 ±â€¯2.8% after transplantation. Histopathology of globe and adnexal tissues showed unique and differential patterns of inflammatory cell infiltration. The ON revealed a neurodegeneration pattern. CONCLUSION: The present study is the first in the literature to establish an experimental model of WET. This model holds significant potential in investigating mechanistic pathways, monitoring strategies or developing management approaches involving ocular viability, immune rejection, and ON regeneration after WET.