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.

Effects of transpositional muscle flaps transfected with vascular endothelial growth factor gene in the treatment of experimental osteomyelitis.

BACKGROUND: Based on the angiogenetic and stimulating effects of bone healing and formation of vascular endothelial growth factor (VEGF), the present study was designed to assess the efficacy of VEGF gene application in the management of experimentally induced osteomyelitis. METHODS: Thirty-two male Sprague Dawley rats were divided into 4 groups, and osteomyelitis was induced in the left tibial bones. Group 1 (n=8) was designated as a control group, and, after the induction of osteomyelitis, no treatment was applied for a period of 4 weeks. Group 2 (n=8) received only antibiotic treatment for 4 weeks following induction of osteomyelitis. In Group 3 (n=8), proximally pedicled gastrocnemius muscle flap was transposed over the osteomyelitic region following induction of osteomyelitis and antibiotic treatment applied for a 4-week period. In Group 4 (n=8), VEGF gene-transfected gastrocnemius muscle flap was transposed over the osteomyelitic region following identical antibiotic regimen applied for a 4-week period. For each group, body temperature, white blood cell (WBC) count, and radiological and histological parameters were evaluated. RESULTS: Body temperature and WBC count remained high in the control group, but returned to normal in Groups 2, 3, and 4 after the third week of treatment. Statistical analysis of the total scores of radiological and histological results revealed significant differences between Groups 1 and 3, Groups 1 and 4, Groups 2 and 3, and Groups 2 and 4 (p<0.05). Regarding radiological parameters of abscess and sequester, and histological parameter of abscess, statistically significant differences were found between Group 4 and the other groups (p<0.05). CONCLUSION: The efficacy of the VEGF gene-transfected muscle flap in the management of experimental osteomyelitis was proven by the results of the present study.

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.

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.

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.

Preservative solution for skeletal muscle biopsy samples.

CONTEXT: Muscle biopsy samples must be frozen with liquid nitrogen immediately after excision and maintained at -80°C until analysis. Because of this requirement for tissue processing, patients with neuromuscular diseases often have to travel to centers with on-site muscle pathology laboratories for muscle biopsy sample excision to ensure that samples are properly preserved. AIM: Here, we developed a preservative solution and examined its protectiveness on striated muscle tissues for a minimum of the length of time that would be required to reach a specific muscle pathology laboratory. MATERIALS AND METHODS: A preservative solution called Kurt-Ozcan (KO) solution was prepared. Eight healthy Sprague-Dawley rats were sacrificed; striated muscle tissue samples were collected and divided into six different groups. Muscle tissue samples were separated into groups for morphological, enzyme histochemical, molecular, and biochemical analysis. STATISTICAL METHOD USED: Chi-square and Kruskal Wallis tests. RESULTS: Samples kept in the KO and University of Wisconsin (UW) solutions exhibited very good morphological scores at 3, 6, and 18 hours, but artificial changes were observed at 24 hours. Similar findings were observed for the evaluated enzyme activities. There were no differences between the control group and the samples kept in the KO or UW solution at 3, 6, and 18 hours for morphological, enzyme histochemical, and biochemical features. The messenger ribonucleic acid (mRNA) of ß-actin gene was protected up to 6 hours in the KO and UW solutions. CONCLUSION: The KO solution protects the morphological, enzyme histochemical, and biochemical features of striated muscle tissue of healthy rats for 18 hours and preserves the mRNA for 6 hours.

Epiphora as the presenting sign of relapsed non-Hodgkin lymphoma in a child with Bruton agammaglobulinemia.

PURPOSE: Primary or secondary infiltration of the lacrimal drainage system by a lymphoid neoplasm is rare in children. Primary immunodeficiencies are characterized by occurrence of unusual malignancies at unexpected locations in the pediatric age group. METHODS: Case report. RESULTS: A 12-year-old boy with a history of Bruton agammaglobulinemia and non-Hodgkin lymphoma (NHL) that primarily originated in the perianal region was referred to our oculoplastics department for persistent epiphora. Computed tomography scan and nasal endoscopy revealed relapse of NHL in the inferior portion of the nasolacrimal duct. Complete remission was achieved with chemotherapy. CONCLUSIONS: Epiphora could be the initial manifestation of a relapse or a recurrence of an underlying malignancy in the pediatric population with predisposing immunodeficiency.

Vas deferens invasion: A neglected issue in the sampling of radical prostatectomy materials.

A radical prostatectomy affects the prostate, bilateral seminal vesicles (SV), and the distal parts of the bilateral vasa deferentia (VD). SV invasion (SVI) is associated with an increased risk of lymph node metastasis and recurrence. However, the significance of VD invasion (VDI), either with or without the involvement of their surgical margins, has not been fully appreciated. We think VDI might have an independent prognostic significance, as does SVI, and should be incorporated into the pathology guidelines and the staging systems of prostatic adenocarcinoma. Our case illustrates this.

Effectiveness of hyperbaric oxygen and ozone applications in tissue healing in generated soft tissue trauma model in rats: an experimental study.

BACKGROUND: Soft tissue trauma is a type of acute traumatic ischemia. We investigated in this study whether the edema, inflammation and ischemia caused by the trauma could be affected positively by hyperbaric oxygen (HBO) and ozone therapy. METHODS: Soft tissue trauma was generated in a total of 63 adult male Sprague-Dawley rats. Subsequently, rats were divided into three groups. The first group was treated with ozone, the second group with HBO, and the third group served as controls. Tissue and blood samples were taken at the end of the procedures. Tissue lipid peroxidation (LPO), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), inducible nitric oxide synthase (iNOS), heme oxygenase (HO)-1, and hypoxia-inducible factor (HIF)-1 levels were detected. Hematoxylin-eosin staining was used to determine the inflammation and edema histopathologically. RESULTS: We also detected HIF-1 activity, which decreases when the oxygen concentration increases, HO-1 activity, which has anti-inflammatory effects, and iNOS activity, which releases in any type of acute case. We determined a statistically significant reduction in iNOS and LPO levels in both the HBO and Ozone groups. A significant decrease in inflammation was detected in both the Ozone and HBO groups compared with the Control group, and a significant decrease in edema was detected in all three groups. CONCLUSION: We think that HBO and Ozone therapy have beneficial effects on biochemical and histopathological findings. Related clinical trials will be helpful in clarifying the effects.

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.

Fibrous pseudotumors of the testis: The balance between sparing the testis and preoperative diagnostic difficulty.

OBJECTIVE: To determine a paradigm that will be helpful for urologists to manage fibrous pseudotumors, which are a very rare condition of the testis. MATERIAL AND METHODS: We retrospectively evaluated the patients with fibrous pseudotumors in our uropathological database from 1995 to 2013. Patients who had tumor markers and ultrosonography (USG) screening before surgery and a final pathology report of a fibrous pseudotumor were included in the study. RESULTS: In total, 838 patients with a testis mass were evaluated. Only 6 of these patients met the inclusion criteria. The mean age was 34 years (min: 20, max: 72). Serum tumor markers were in the normal range for all patients. The scrotal tumors were not clearly related to the testis parenchyma, and the radiologists could not definitively determine the nature of the masses (benign or malignant). A concomitant hydrocele was detected in 2 patients. After inguinal exploration, radical orchiectomy was performed in one patient with an ipsilateral atrophic testis, and biopsies were taken from the lesions for frozen section analysis in the other 5 patients. Pathologists reported benign tumors for all of these patients based on the frozen sections, and testicular sparing surgery was then performed in these 5 patients. CONCLUSION: If scrotal tumors are detected by ultrasonography in patients with normal tumor markers, and the tumor cannot be clear distinguished from the testis, these patients might have a fibrous pseudotumor, and organ-sparing surgery can be performed on these patients.

Lumbar epidural varix mimicking perineural cyst.

Lumbar epidural varices are rare and usually mimick lumbar disc herniations. Back pain and radiculopathy are the main symptoms of lumbar epidural varices. Perineural cysts are radiologically different lesions and should not be confused with epidural varix. A 36-year-old male patient presented to us with right leg pain. The magnetic resonance imaging revealed a cystic lesion at S1 level that was compressing the right root, and was interpreted as a perineural cyst. The patient underwent surgery via right L5 and S1 hemilaminectomy, and the lesion was coagulated and removed. The histopathological diagnosis was epidural varix. The patient was clinically improved and the follow-up magnetic resonance imaging showed the absence of the lesion. Lumbar epidural varix should be kept in mind in the differential diagnosis of the cystic lesions which compress the spinal roots.

Oral epithelial barrier function and the role of nuclear factor kappa-ß pathway in the pathogenesis of aphthous ulceration.

BACKGROUND/AIMS: The normal oral mucosa is usually tolerant to its special microenvironment. Epithelial integrity and a wellmanaged immune system are important in sustaining harmony. A close look at the role played by adaptive immunity during recurrent aphthous ulcerations may throw some light into the pathogenesis. MATERIALS AND METHODS: In this report, we provide a concise review of oral epithelial barrier function and present data on the possible pathogenetic mechanism of aphthous ulceration using immunohistocemical signs of nuclear factor kappa beta pathway activation on fourteen cases of mucosal aphthous ulcerations. RESULTS: We strongly support the hypothesis that oral aphthous ulcerations develop as a result of loss of epithelial barrier function and that nuclear factor kappa beta signaling pathway seems to be involved in this type of injury. CONCLUSION: Interventions that strengthen the mucosal barrier function or modulate inappropriate activation of nuclear factor kappa beta signaling pathway can be considered in the treatment of oral aphthous ulcerations.

Nuclear morphometry in the differential diagnosis of eosinophilic renal epithelial tumors.

OBJECTIVE: To assess the potential contribution of nuclear morphometry to the differential diagnosis of renal epithelial tumors with eosinophilic cytoplasm, including chromophobe renal cell carcinoma (ChRCC), the eosinophilic variant of clear cell renal cell carcinoma (EoRCC), and oncocytoma. STUDY DESIGN: A total of 24 tumor tissue samples diagnosed as ChRCC, the EoRCC, or oncocytoma constituted our series. Eight geometric features such as nuclear area, nuclear perimeter, and circular form factor were measured and compared among the groups. RESULT: On the basis of nuclear morphometry, measurements of eight geometric features significantly differ among these problematic eosinophilic renal epithelial neoplasms (p < 0.005). CONCLUSION: Because of their different biologic behaviors, the exact discrimination of the renal epithelial tumors with eosinophilic cytoplasm is crucial. However, this distinction can sometimes be problematic even for highly experienced pathologists. Our results suggest that the morphometric nuclear shape descriptors may be used as an ancillary method in their differential diagnosis.

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.