Testicular-sparing excision for benign paratesticular IgG4-related disease

A 28-year-old male was referred by his local general practitioner due to recurrence of painful right scrotal mass, first noted 8 years prior. The mass was further characterised with ultrasound and then was locally excised via an inguinal approach, sparing the testicle, without any postoperative complication. Immunoperoxidase staining of the excised lesion confirmed paratesticular IgG4-related disease.

Basal cell carcinoma developed from an epidermal cyst: A case report and review of the literature

Cutaneous epidermal cysts are common benign cysts derived from the epidermis or epithelium of hair follicles, and malignancy originating from epidermal cysts is uncommon. When a cutaneous epidermal cyst turns malignant, it is mostly squamous cell carcinoma, and basal cell carcinoma is rare. We present the case of a 58-year-old man with basal cell carcinoma originating from an epidermal cyst on the buttocks. Histopathological analysis with hematoxylin- eosin (H&E) staining showed the presence of the cyst, the wall of which was partially replaced by a malignant tumor. Moreover, the cyst was filled with malignant tumor cells in some areas. The tumor cells were basaloid, and extended through the dermis down to the subcutaneous tissue in a solid pattern. Immunoperoxidase staining for Ber-EP4 was positive. To our knowledge, there are 11 reported cases of BCC originating from epidermal cysts in English, and we reviewed these reports and examined potential trends. We estimate that some longstanding epidermal cysts may have the potential of malignant transformation, and chronic and repeated irritation might trigger malignancy from epidermal cysts. We emphasize that benignlooking cystic lesions showing progressive growth should be examined histopathologically.

​Dermal Response to Experimental Orfvirus (ORFV) Infection in Goats, Mice and Rabbit

Background: During a study on the outbreak of orf in goats, it was intended to study the disease transmissibility in different hosts from field samples and ascertain the infective potential of the agent in laboratory animals compared to goats. Methods: Cutaneous clinical materials from orf virus (ORFV) infected goats was used to experimentally infect naive goats, rabbits and mice and ascertain its infective potential and transmissibility in different hosts. The processed inoculum was applied topically to mimic a natural transmission through injured skin. Regular skin biopsies were taken that revealed characteristic macroscopic and microscopic lesions typical of orf. Result: Virus inoculum applied on abraded skin in goats successfully established the lesions of orf. A parallel inoculation in rabbit and mice could not successfully reproduce the disease in these unnatural hosts beyond a subtle vesicular stage on 3 dpi with subsequent healing by 7 dpi. The lesions in goats regressed spontaneously by 28 days post-infection (dpi). Intracytoplasmic inclusions were associated only in the vesicular stage. Immunopathological progression was observed by immunoperoxidase staining of CD4+ and CD8+ T-cells which were found to appear by day 5 in the dermis and became more abundant and distributed by day 8, but subsequently reduced in number by 15 dpi. CD4+ cells were found to be more numerous and widespread. Viral antigen in tissues could be demonstrated by 4 dpi by immunohistological methods that increased in signal intensity progressively and disappear by 28 dpi. Similarly, viral nucleic acid in the skin could be detected on day 8 dpi but not on 28 dpi by PCR. The present experiment depicts the ease of disease transmissibility through traumatized skin in the primary hosts, but establishment in unnatural hosts may not be readily achieved. The infection was self-limiting with possibly no virus latency as indicated by immunofluorescence and PCR studies.

A Rare Case of Metastatic Transformation to Pituitary Carcinoma

Background: Pituitary carcinomas are rare and comprise less than 0.5% of pituitary tumors and frequently arise from previously resected and/or radiated infiltrating adenomas.1 Radiation therapy is used to prevent regrowth but does not have significant data supporting improved prognosis.2 True pituitary carcinomas requires the presence of craniospinal and/or systemic metastases. Here, we present a case of benign pituitary adenoma that had progressed to a metastatic tumor and responded to radiation therapy. Clinical Case: Our patient was first diagnosed in 1982 with a pituitary mass after undergoing an MRI due to right eye blindness. He ultimately had a transcranial resection of the tumor and post-operative external radiation to residual tumor. Final pathology was consistent with a nonsecretory tumor. Additionally, he was diagnosed with panhypopituitarism and he was treated accordingly. Due to barriers of patient adherence, only one post-operative MRI was obtained, and he was lost to endocrine follow up. In 2007, he was again referred to endocrinology for new vision loss in the left eye. An MRI revealed a 3.0 x 3.8 x 4.4 cm tumor that compressed the optic chiasm. Additionally, multiple lesions were seen within the dural and leptomeningeal extra-axial areas including the largest left frontal lesion of 2.4 cm and a 1.7 cm lesion posterior to the cord within the foramen magnum. The patient underwent endonasal transsphenoidal resection of the sellar mass and biopsy of the frontal lesion in October 2007. Pathology showed pituitary adenocarcinoma in both the sellar and frontal lesions and immunoperoxidase staining was negative for all hormones. He completed salvage radiation to the sellar and craniospinal lesions in December 2007. MRI scans completed 7 months later showed stable residual pituitary and metastatic lesions. The patient received 6-month MRI screening exams for the first 2 years and then annually thereafter, as well as PET scanning, without significant progression or new metastatic lesions. Conclusion: We present a case of benign pituitary adenoma which transformed following resection to a malignant tumor with metastatic lesions in the brain and spinal cord that was treated with radiation with relative success. Despite the often-aggressive nature of these metastatic tumors, the patient has experienced a successful treatment course with minimal side effects and without progression 13 years after initial treatment of the metastatic pituitary carcinoma.

Factors Affecting The Indirect Immunoperoxidase Staining Of Infectious Bursal Disease Virus Antigens In Cells

THE application of the Immunoperoxidase (IP) antibody technique for the definitive diagnosis of Infectious Bursal Disease (IBD) in chickens has earlier been described (Okeke and Lang, 1982). During that study it was noted that the Indirect method of IP was more sensitive in the demonstration of IBD viral antigens than the direct method. Consequently, more formation was sought on the specific technical parameter and the consequences that resulted by deviations from the codified procedures employed in the Indirect technique with the aim of obtaining an even better result. Studies of pH, serum dilutions, serum incubation time, conjugate dilutions, conjugate incubation time and developing time were conducted to find out what Influence such factors could have on the final staining product.

Infectivity and viability of dengue virus infected hepatocytes cocultured with peripheral blood mononuclear cells from a healthy subject

BACKGROUND Dengue virus (DENV) can infect and replicate in monocytes, resulting in antibody-dependent enhancement. The liver is the main target of DENV, and the infection mechanisms of DENV include direct cytopathic effects (CPEs) of the virus, mitochondrial dysfunction, and effect of cellular and humoral immune factors in the liver. This study was aimed to explore the infectivity of DENV and viability of human hepatocytes using Huh 7it-1 cells cocultured with peripheral blood mononuclear cells (PBMCs). METHODS Huh 7it-1 cells were infected with dengue virus serotype-2 (DENV-2) New Guinea C strain at multiplicity of infection of 0.5 and 1 FFU/cell, and cocultured in vitro with and without adherent PBMCs. The infectivity of DENV was assessed by immunoperoxidase staining. The viability of Huh 7it-1 cells was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, a tetrazole) assay and trypan blue staining. Data were statistically analyzed by Shapiro–Wilk and analysis of variance for normality significances. RESULTS The result showed that addition of PBMCs to DENV-2 infected Huh 7it-1 cells decreased the infectivity of DENV (15–37%). DENV-2 infection decreased the viability of Huh 7it-1 cells (15.5–20.8%). Despite the decrease in infectivity of DENV, the addition of PBMCs increased the Huh 7it-1 cells viability (4.5–10.2%). CONCLUSIONS Addition of PBMCs to Huh 7it-1 cells that are infected with DENV-2 decreased the infectivity of DENV and increased Huh 7it-1 cells viability.

127 Chemokine receptor 2 (CCR2) is expressed in growing oocytes, and its deficiency affects follicular activation and long-term female fertility in mice

Chemokine receptor 2 (CCR2) has important functions in several biological processes, including activation of PI3K/Akt/mTOR signalling, a key pathway in follicular activation. However, there is no report about the role of CCR2 in ovarian follicular physiology. The objectives of this study were (1) to immunolocalize CCR2 in mice ovaries and (2) to evaluate the effects of CCR2 deficiency on follicular growth during adult life and aging. A total of 74 C57Bl/6 (wild type, WT) and 68 B6.129S4-Ccr2tm1Ifc/J (CCR2−/− (knockout)) female mice were used. For objective 1, ovaries were collected from WT mice at 1.5 months old (m.o.), fixed in 4% PFA, embedded in paraffin, and used for immunoperoxidase staining with an anti-CCR2 antibody [EPR19698] (ab222496; 1:200) and using anti-rabbit IgG (Ab6721, 1:100) as a secondary antibody. Also, MII oocytes from oviducts of superovulated WT mice were processed with the same primary antibody for immunofluorescence. For objective 2, body and ovarian weight were evaluated. Follicle populations were assessed in WT and CCR2−/− mice at 1.5, 2.5, 6, 10, and 12m.o., by serial sectioning; the total follicle population was counted in every third section in the whole ovary. Additionally, ovarian total RNA isolation was performed from WT and CCR2−/−. Real-time PCR was used to evaluate differential gene expression according to standard protocols, using primers for Bax, Casp3, Bcl2, Fshr, and B-actin (endogenous control). The data were analysed using the GraphPad Prism Software, using t-test, and a P-value of 0.05 was considered as significant. Localization of CCR2 was observed exclusively on the membrane and cytoplasm of growing oocytes in primary, secondary, antral, and atretic follicles, as well as on ovulated MII oocytes membrane and cytoplasm. Body and ovarian weight were similar between WT and CCR2−/− mice. At 1.5m.o., CCR2−/− mice had more primordial follicles and fewer primary and secondary follicles compared with WT (P<0.05), whereas there was no difference in the antral follicle populations. Follicular activation (primordial to primary transition) and atresia rates were decreased in CCR2−/− (P<0.05) at 1.5m.o. A downregulation of pro-apoptotic genes (Bax and Casp3) was observed on CCR2−/− (P<0.05), while anti-apoptotic Bcl2 was upregulated (P<0.05) compared with WT in 1.5-m.o. animals. A larger ovarian follicular reserve at 1.5, 2.5, and 6m.o., but not at 10 or 12m.o., was observed in CCR2−/− mice. At 6 to 12m.o., CCR2−/− ovulated more (P<0.05) MII oocytes than WT. Altogether, these data may suggest that CCR2 plays an important role in the regulation of ovarian follicular reserve mobilization, potentially affecting reproductive lifespan. Financial support was provided by Fapemig and CNPq.

Subcutaneous panniculitis-like T-Cell lymphoma: A mixed diagnostic approach to diagnosing a vague clinical picture

Subcutaneous panniculitis-like T-Cell lymphoma (SPTCL) is a rare subtype of cutaneous T-cell lymphoma, and it has been associated with a range of clinical symptoms from mild to severe. Most commonly, this disease is described as following a slowly progressing course, associated with vague constitutional symptoms and good prognosis. This case report describes the clinical presentation and findings of SPTCL in a 31 year old female and describes the challenges of recognizing and properly diagnosing this disease. SPTCL has been described as a mimicker of other, more common and nonmalignant diseases of the skin, such as lupus panniculitis. This report highlights a variety of specific tests including immunohistochemical and immunoperoxidase staining, as well as genotypic analysis of T-cell receptors, that were effective in combination in isolating this diagnosis. Moreover, choice of treatment for these patients can be challenging, as an array of interventions have been described in past cases to treat SPTCL. This report recognizes the efficacy of a treatment course that included a six-cycle course of combined chemotherapy (vincristine, doxurubicine, cyclophosphamide, and prednisone, also known as CHOP) followed by weekly methotrexate and PET scan surveillance for two years. With both initial and maintenance therapy, this patient showed excellence response evidenced by a progressive decrease in metabolic activity of malignant lesions, lack of new lesions, and remaining without symptoms. While this disease is rare, it is important to include SPTCL in the differential when considering patients with a panniculitic picture.

Lymphatic vessels identified in failed corneal transplants with neovascularisation

BackgroundCorneal transplant failure with neovascularisation is a leading indication for full-thickness grafts in patients. Lymphangiogenesis is implicated in the pathology of graft failure, and here we systematically evaluate failed human corneal transplants with neovascularisation for the presence of lymphatic vessels.MethodsNine failed grafts with neovascularisation, based on H&E staining with subsequent immunoperoxidase staining for CD31, a blood vessel marker, were selected. Lymphatics were investigated by immunohistochemical and immunofluorescence approaches using podoplanin as a lymphatic marker. In two of nine cases, fluorescence in situ hybridisation (FISH) was used for detection of lymphatic mRNAs including podoplanin, VEGFR-3 and LYVE-1. All immunofluorescence and FISH samples were compared with positive and negative controls and visualised by confocal microscopy.ResultsCorneal neovascularisation was established in all cases by H&E and further confirmed by CD31 immunoreactive profiles. Immunohistochemistry for the podoplanin antibody was positive in all cases and showed morphologies ranging from distinct luminal structures to elongated profiles. Simultaneous immunofluorescence using CD31 and podoplanin showed lymphatic vessels distinct from blood vessels. Podoplanin immunofluorescence was noted in seven of nine cases and revealed clear lumina of varying sizes, in addition to lumen-like and elongated profiles. The presence of lymphatic mRNA was confirmed by FISH studies using a combination of at least two of podoplanin, VEGFR-3 and LYVE-1 mRNAs.ConclusionsThe consistent finding of lymphatic vessels in failed grafts with neovascularisation implicates them in the pathogenesis of corneal transplant failure, and points to the lymphatics as a potential new therapeutic target.