Large-cell calcifying Sertoli cell tumour with macrocalcification in partially resected testis of young adult patient.

INTRODUCTION: There are less than 100 cases of Large-cell calcifying Sertoli cell tumour (LCCSCT) reported in English literature. Most of them are benign, bilateral and affect paediatric population. Malignant cases occur in older patients. LCCSCT is often associated with Carney complex or Peutz-Jaghers syndrome. We present the clinicopathological features of a young adult, with unilateral "stone-like" LCCSCT, without changes in hormonal status and no clinical characteristics of noted genetic disorders. CASE REPORT: A 24-year-old male presented with painless hardening of the right testis. There was no gynaecomastia, and serum levels of human chorionic gonadotropin and α-fetoprotein were normal. Ultrasound depicted hyperechogenic, clearly demarcated intratesticular lesion. Partial orchiectomy was performed. Macroscopically, tumour appeared as almost entirely calcified round mass, measuring 10 mm. Histopathological evaluation showed well-circumscribed, unencapsulated tumour composed of massive calcified geographic formations, surrounded with tumour cells. Neoplastic cells were large, polygonal, with abundant eosinophilic cytoplasm, and formed irregular cords, pseudo tubular structures, and nests in a fibrous and myxoid stroma, surrounded with lymphocytes. Other forms of calcification were also present: Needle-like deposits and lamellar, mulberry-like structures. There was no necrosis, mitotic activity and nuclear pleomorphism. Immunohistochemical study was positive for inhibin α and negative for Melan A, EMA, synaptophysin, chromogranin and AFP. DISCUSSION: LCCSCT needs to be differentiated from other, more frequent, sex cord stromal tumours. Clinical and genetical evaluation of these patients had to be performed, due to connection of LCCSCT with genetic abnormalities. In evidently benign cases, organ-sparing surgery should be considered for younger patients, followed by long term follow-up.

The safety and feasibility of probiotics in children and adolescents undergoing hematopoietic cell transplantation.

Hematopoietic cell transplantation (HCT) has become a standard treatment for many adult and pediatric conditions. Emerging evidence suggests that perturbations in the microbiota diversity increase recipients' susceptibilities to gut-mediated conditions such as diarrhea, infection and acute GvHD. Probiotics preserve the microbiota and may minimize the risk of developing a gut-mediated condition; however, their safety has not been evaluated in the setting of HCT. We evaluated the safety and feasibility of the probiotic, Lactobacillus plantarum (LBP), in children and adolescents undergoing allogeneic HCT. Participants received once-daily supplementation with LBP beginning on day -8 or -7 and continued until day +14. Outcomes were compliance with daily administration and incidence of LBP bacteremia. Administration of LBP was feasible with 97% (30/31, 95% confidence interval (CI) 83-100%) of children receiving at least 50% of the probiotic dose (median 97%; range 50-100%). We did not observe any case of LBP bacteremia (0% (0/30) with 95% CI 0-12%). There were not any unexpected adverse events related to LBP. Our study provides preliminary evidence that administration of LBP is safe and feasible in children and adolescents undergoing HCT. Future steps include the conduct of an approved randomized, controlled trial through Children's Oncology Group.

Baerveldt tube implantation following failed deep sclerectomy versus repeat deep sclerectomy.

PURPOSE: To compare the surgical outcomes of repeat deep sclerectomy (DS) and the Baerveldt glaucoma implant (BGI) in eyes with failed primary deep sclerectomy. DESIGN: A retrospective comparative case-control study. METHODS: Fifty-eight eyes of 56 glaucoma patients with previously failed DS underwent BGI (group BGI) and 58 eyes of 55 patients underwent repeat DS (group DS) at a tertiary referral centre. Visual acuity, intraocular pressure (IOP), number of glaucoma medications, surgical failure rates and complication rates were compared between groups. Surgical failure was defined as loss of IOP control, loss of light perception, or need for further glaucoma surgery. RESULTS: Baseline demographics were similar between groups. Preoperatively, median IOP was lower in the DS than the BGI group (19 mmHg versus 21 mmHg, p = 0.10). Postoperatively at year 1, median IOP was significantly higher in the DS than BGI group (14 mmHg versus 11 mmHg, p = 0.02). There were no differences between the DS and BGI groups in mean number of medications preoperatively (2.3 versus 2.6) or postoperatively (1.3 versus 1.1). Complication rates were significantly higher in the DS group [41 % (n = 24) versus 14 % (n = 8); p = 0.01]. The failure rate at 1 year was higher in eyes with repeat DS than in eyes with BGI (30 vs. 21 %, respectively; p = 0.07). CONCLUSIONS: Baerveldt implants were more effective in lowering IOP and resulted in significantly fewer complications than repeat deep sclerectomy in eyes with previously failed deep sclerectomy.

A device for the application of uniaxial strain to single crystal samples for use in synchrotron radiation experiments.

We present the design, construction, and testing of a straining device compatible with many different synchrotron radiation techniques, in a wide range of experimental environments (including low temperature, high field and ultra-high vacuum). The device has been tested by X-ray diffraction on single crystal samples of quasi-one-dimensional Cs2Mo6Se6 and K2Mo6Se6, in which microscopic strains up to a Δc/c = 0.12% ± 0.01% change in the c lattice parameters have been achieved. We have also used the device in an inelastic X-ray scattering experiment, to probe the strain-dependent speed of sound ν along the c axis. A reduction Δν/ν of up to -3.8% was obtained at a strain of Δc/c = 0.25% in K2Mo6Se6.

Pseudophakic Retinal Detachment Surgery by 23 G Vitrectomy using Slit-Lamp and Non-Contact 90 D Lens.

BACKGROUND: The purpose of this study is to report the anatomic and functional results of primary 23 G vitrectomy using slit-lamp and non-contact 90 D lens for the treatment of pseudophakic rhegmatogenous retinal detachment. PATIENTS AND METHODS: Pseudophakic eyes were operated by 23 G vitrectomy using slit-lamp and non-contact 90 D lens, internal subretinal fluid drainage, cryopexy and internal gas tamponade. The preoperative and postoperative characteristics were analysed. Main outcome measures were anatomic success rates after initial surgical intervention and after reoperation for primary failures, visual outcome at the last follow-up visit, and complications. RESULTS: 46 pseudophakic eyes were included in this retrospective study (October 2013- January 2014). In 40 cases, sulfur hexafluoride 23% gastamponade was used, silicone oil in 6 cases (13%). The retina was reattached successfully after a single surgery in 44 eyes (96%). Recurrence of retinal detachment occurred in 2 eyes. Final anatomic reattachment was obtained in 100% after a second operation. Silicone oil was removed in all eyes. Visual acuity improved significantly from logMAR 0 (IQR 0-0.9) to logMAR 0 (IQR 0-0.2) (p<0.005). CONCLUSIONS: Primary 23 G vitrectomy using slit-lamp and non contact 90 D lens for the treatment of pseudophakic rhegmatogenous retinal detachment provides a high anatomic and functional success rate and is associated with few complications.

Multicolor counterstaining for immunohistochemistry -- a modified Movat's pentachrome.

The application of "histochemical" staining procedures has been substantially replaced by immunostaining of specific molecular tissue components. The limited range of colors resulting from routine immunohistochemistry, however, can limit assessment of the general microscopic tissue organization. Consequently we have adapted a polychromatic histochemical counterstaining procedure based on Movat's pentachrome staining sequence for use with immunohistochemical procedures. The value of Movat's original method when applied as an immunohistochemical counterstain is limited by its use of iron hematoxylin and by fact that the resulting color combination is difficult to distinguish from the colors of routine immunohistochemical staining. Our variant pentachrome stains the same tissue components as Movat's stain; however, owing to a modification of the acid fuchsin staining step, it provides a strong color contrast with the reaction product resulting from immunostaining using diaminobenzidine as the chromogen. Multicolor counterstaining for immunohistochemistry offers a new approach to tissue analysis, especially when stromal-epithelial relations of normal and neoplastic tissues are considered.

FOXP3 expression following bone marrow transplantation for IPEX syndrome after reduced-intensity conditioning.

The objective of this study is to determine if immune reconstitution of FOXP3+ T regulatory cells correlates with clinical improvement of IPEX syndrome following allogeneic hematopoietic stem cell transplant. An 8-months-old male infant with a mutation in the polyadenylation site of FOXP3 gene, absence of FOXP3 protein expression and clinical manifestations of IPEX syndrome, including eczema, colitis, failure to thrive, TPN requirement, and elevated serum IgE, underwent matched unrelated hematopoietic stem cell transplant. After reduced-intensity conditioning with alemtuzumab followed by fludarabine and melphalan the patient's neutrophils engrafted day +15 and platelets day +29. Patient was a full donor chimera day +28 and +60. Intracellular FOXP3 protein expression in CD4+ T cells was absent pre-HSCT. After transplantation, percentage CD4+ T cells expressing FOXP3+CD25 bright phenotype quickly increased from 4.5 (day +29) to 23% (day +90) and continued in this trend. Foxp3 mRNA expression confirmed flow cytometry data. Serum IgE levels decreased from 5,000 IU/ml pre-transplant to 6 IU/ml on day +90, eczema resolved, and secretory diarrhea and feeding intolerance improved. T regulatory cell reconstitution is evident soon after HSCT following reduced-intensity conditioning correlating with development of full donor chimerism. Increased FOXP3 expression correlates with correction of clinical and laboratory manifestations of IPEX syndrome providing direct evidence that HSCT is a curative procedure for this disorder.

Hematopoietic stem cell transplantation for pediatric patients with primary immunodeficiency diseases at All Children's Hospital/University of South Florida.

We retrospectively analyzed the transplantation outcomes of 31 patients with primary immunodeficiency diseases treated at our center (All Children's Hospital, University of South Florida) since its inception in 1986. The primary immune diseases included severe combined immunodeficiency, Wiscott-Aldrich syndrome, X-linked hyper-IgM syndrome, and chronic granulomatous disease. The age of the patient's at the time of transplant ranged from 1 month to 19 years, and conditioning regimens varied based on the patients underlying disease. In 23 patients, the graft source was bone marrow, 4 patients received umbilical cord blood grafts and 4 patients received peripheral blood stem cell grafts. Better survival rates were observed in those patients transplanted at a younger age and free of infections, demonstrating that transplantation at an early age before significant infections, autoimmune manifestation and malignant transformation have occurred is beneficial.

Feasibility study of preemptive withdrawal of immunosuppression based on chimerism testing in children undergoing myeloablative allogeneic transplantation for hematologic malignancies.

An increasing percentage of autologous cells (increasing chimerism) in the whole blood (WB) chimerism test following allogeneic transplant is related to a very high risk of relapse. Preemptive immunotherapy may decrease the risk of relapse in some patients. Our prospective multi-institutional study evaluated the feasibility of longitudinal chimerism testing in a central laboratory, compared WB, CD3+ and leukemia-specific lineage chimerism in patients with a variety of hematologic malignancies, and evaluated the feasibility of fast withdrawal of immunosuppression based on WB chimerism results. Centralized chimerism testing was feasible and showed low interassay variability. Increasing mixed chimerism (MC) in WB was not useful as a predictor of relapse in our study. The presence of full donor chimerism in WB, CD3+ and leukemia-specific lineages on all measurements was related to a significantly lower risk of relapse than the presence of MC in either subset (11 vs 71%, respectively; P=0.03). Increasing host chimerism in leukemia-specific lineage heralds relapse, but it was not detected early enough to allow immunotherapy. Further studies correlating lineage-specific chimerism and minimal residual disease are required. The goal of preemptive immunotherapy should be to achieve full donor chimerism in WB in CD3+ and leukemia-specific lineages.

Follicular dendritic cell sarcoma in the lymph nodes of the neck.

Follicular dendritic cell sarcoma (FDCS) is a rare tumor, derived from antigen-presenting cells that normally form meshworks in lymphoid follicles. FDCS in general recapitulate the immunophenotypic profile of follicular dendritic cells (FDCs). This tumor affects mainly the lymph nodes, but may arise at a variety of extranodal sites in about one third of cases. We present a case of FDCS in a 52-year-old woman with left neck mass. Histologically, the tumor was composed of oval-to-spindle cells arranged in a fascicular and storiform pattern. Immunohistochemically, the tumor cells were positive for CD21, CD35, and to a lesser extent for EMA and vimentin (Fig. 3, Ref. 14). Full Text (Free, PDF) www.bmj.sk.

Cathepsin D-related disease-free interval in pT1 primary breast carcinomas: a pilot study.

In order to address the heterogeneity of the pT1 breast cancer stages, we have been examining the natural and the clinical course of disease in relation to cathepsin D expression, as a molecular marker for the tumor progression that leads to metastasis. The original aim of our pilot study was to determine whether it was possible to distinguish high-risk from low-risk patients, on the basis of nonestrogen- vs. estrogen-regulated cathepsin D expression. Our results showed that estrogen-regulated cathepsin D expression could be useful as surrogate marker of node-positive status. Further, during the natural course of disease, none of 7 pT1N0 patients with tumors bearing nonestrogen-regulated cathepsin D expression developed metastasis. During the clinical course of disease, nonestrogen-regulated cathepsin D expression defined low-risk while estrogen-regulated cathepsin D expression defined high-risk pT1N+ subgroup of patients. Although there is no consensus with respect to metastasis-related prognostic value of cathepsin D expression, our pilot study implies its prognostic value in pT1 breast cancer patients and supports the hypothesis that cathepsin D may promote metastasis in this early stage of disease.

A membrane-proximal basic domain and cysteine cluster in the C-terminal tail of CCR5 constitute a bipartite motif critical for cell surface expression.

We examined the structural requirements for cell surface expression, signaling, and human immunodeficiency virus co-receptor activity for the chemokine receptor, CCR5. Serial C-terminal truncation of CCR5 resulted in progressive loss of cell surface expression; mutants truncated at the 317th position and shorter were not detected at the cell surface. Alanine substitution of basic residues in the membrane-proximal domain (residues 314-322) in the context of a full-length C-tail resulted in severe reduction in surface expression. C-terminal truncation that excised the three cysteines in this domain reduced surface expression, but further truncation of upstream basic residue(s) abolished surface expression. Substituting the carboxyl-terminal domain of CXCR4 for that of CCR5 failed to rectify the trafficking defect of the tailless CCR5. In contrast, tailless CXCR4 or a CXCR4 chimera that exchanged the native cytoplasmic domain for that of wild type CCR5 was expressed at the cell surface. Deletion mutants that expressed at the cell surface responded to chemokine stimulation and mediated human immunodeficiency virus entry. Substitution of all serine and threonine residues in the C-terminal tail of CCR5 abolished chemokine-mediated receptor phosphorylation but preserved downstream signaling (Ca(2+) flux), while substitutions of tyrosine residues in the C-tail affected neither phenotype. CCR5 mutants that failed to traffic to the plasma membrane did not exhibit obvious changes in metabolic turnover and were retained in the Golgi or pre-Golgi compartments(s). Thus, the basic domain (-KHIAKRF-) and the cysteine cluster (-CKCC-) in the C-terminal tail of CCR5 function cooperatively for optimal surface expression.

Exchange of the basic domain of human immunodeficiency virus type 1 Rev for a polyarginine stretch expands the RNA binding specificity, and a minimal arginine cluster is required for optimal RRE RNA binding affinity, nuclear accumulation, and trans-activation.

The Rev regulatory protein of human immunodeficiency virus (HIV) facilitates the nuclear export of unspliced and partially spliced HIV RNAs. Using a Rev:MS2 phage coat protein fusion that could be targeted to bind and activate the Rev-responsive element (RRE) RNA or heterologous MS2 phage operator RNA, we analyzed the role(s) of the arginine-rich RNA binding domain in RNA binding and transactivation. The arginine-rich domain could be functionally replaced by a stretch of nine arginines. However, polyarginine substitutions expanded the RNA binding specificity of the resultant mutant Rev protein. Polyarginine insertions in place of residues 24 to 60 that excised the RNA binding and oligomerization domains of Rev preserved the activation for MS2 RNA, but not for the RRE. A nine-arginine insertion outside of the natural context of the Rev nuclear localization signal domain was incompatible with activation of either RNA target. Insertions of fewer than eight arginines impaired RRE activation. Interrupted lysine clusters and disruption of the arginine stretch with lysine or neutral residues resulted in a similar phenotype. Some of these mutants with a null phenotype for RRE activated the heterologous MS2 RNA target. Under steady-state conditions, mutants that preserved the Rev response for RRE RNA localized to the nuclei; those with poor or no Rev response accumulated mostly in the cytoplasm. Many of the cytoplasmically resident derivatives became nuclear when leptomycin B (LMB) treatment inhibited nuclear export of nuclear export signal-containing proteins. Mutants that had a null activation potential for either RNA target were particularly resistant to LMB treatment. Abbreviated nuclear residence times and differences in RRE binding affinity may have compromised their activation potential for RRE. High-affinity binding to MS2 RNA through the intact coat protein was sufficient to overcome the short nuclear residence times and to facilitate MS2 activation by some derivatives.

The mitochondrial phosphoglyceroyl-ATP-containing polymer, purinogen, is unchanged by cardiac ischaemia and reperfusion but may function in the regulation of free intracellular inorganic phosphate concentrations.

Previous work in our laboratory demonstrating large unexplained systematic variations in the heart contents of free adenine nucleotides led us to propose the existence of some unrecognised sequestered form and thence to the purification of very labile acid-insoluble oligomers which we characterised as oligo[3-phospho-glyceroyl-gamma-triphospho(5')adenosine(3')] , abbreviated to (PG-ATP)n. More recently, we provided evidence that these oligomers appear to be the end chains of a complex polymer located in the mitochondrial intermembrane space of a number of rat tissues. We called this polymer purinogen and devised a means of assaying it quantitively [Patel, B., Sarcina, M. & Mowbray, J. (1994) Eur. J. Biochem. 220, 663-669]. Here we report measurements of purinogen in perfused hearts subjected to moderate and severe global ischaemia and reperfusion. Measurements of tissue and perfusate nucleotides, nucleosides and purine degradation products demonstrate that ischaemia led to the augmentation of the free nucleotide content by up to 30% and its re-sequestration on reperfusion in reversible but not in irreversible ischaemia. The purinogen content was unchanged by ischaemia or reperfusion implying the existence of some other unidentified storage pool. By contrast, glucose addition to glycolytically deprived hearts or removal of Pi from perfusion medium, conditions which might be expected to alter demand for intracellular Pi, led to the quantitative transfer of nucleotides between phosphate-rich purinogen and free nucleotides. The possibility that purinogen may act as a rapidly accessible reservoir of intracellular inorganic phosphate is discussed.

The Ig heavy chain 3' end confers a posttranscriptional processing advantage to Bcl-2-IgH fusion RNA in t(14;18) lymphoma.

The chromosomal translocation t(14;18) in lymphoma leads to an overproduction of the Bcl-2 protein on the basis of increased Bcl-2 mRNA levels. Whereas the juxtaposition of Bcl-2 with the Ig heavy chain locus causes a transcriptional activation, 70% of the lymphomas also produce Bcl-2-Ig fusion RNAs with Ig 3' ends. Using S1 nuclease protection assays that can discriminate between nuclear RNA precursors and spliced mRNA, we found that the fusion RNAs in t(14;18) cell lines exhibit an additional posttranscriptional processing advantage. Transfection experiments with artificial genes containing various Bcl-2 or Ig 3' ends show that this effect is (1) related to RNA splicing and/or nucleocytoplasmic transport; (2) independent of transcriptional activation by the heavy chain enhancer; (3) dependent on the presence of the JH-CH and C-gamma1 Ig introns; and (4) tissue specific for B cells. This constitutes a novel mechanism of oncogene deregulation unrelated to transcriptional activation or half-life prolongation. The data further support the existence of a tissue-specific posttranscriptional pathway of Ig regulation in B cells.

Competitive CBFbeta/MYH11 reverse-transcriptase polymerase chain reaction for quantitative assessment of minimal residual disease during postremission therapy in acute myeloid leukemia with inversion(16): a pilot study.

PURPOSE: (1) Quantification of minimal residual disease (MRD) by competitive CBFbeta/MYH11 reverse-transcriptase polymerase chain reaction (RT-PCR) in patients with acute myeloid leukemia (AML) and inversion(16) [inv(16)] during postremission therapy, (2) comparison of this method with conventional two-step RT-PCR, and (3) evaluation of a potential prognostic value. PATIENTS AND METHODS: MRD of six consecutive adult patients with AML and inv(16)(p13;q22) or t(16;16)(p13;q22) who entered complete remission (CR) was monitored by competitive CBFbeta/MYH11 RT-PCR in their bone marrow (BM) during postremission therapy with high-dose cytarabine (HiDAC) or after BM transplantation with a matched unrelated-donor marrow (MUD-BMT) during an observation period of 4.5 to 27 months after initiation of treatment. RESULTS: Competitive PCR showed a gradual decline by at least 4 orders of magnitude after 7 to 9 months in patients in continuous CR (CCR), while one patient who relapsed after 13.5 months only achieved a reduction by 2 orders of magnitude at the end of consolidation therapy. A rapid decrease below the detection limit was observed within 1 month in two patients after MUD-BMT. A temporary reappearance of molecular MRD was observed in these patients during immunosuppression for graft-versus-host disease (GvHD). After reduction of immunosuppression, the level of MRD dropped again below the PCR detection limit. Molecular monitoring by conventional two-step RT-PCR yielded comparable results only when multiple assays per time point were performed, while single-assay RT-PCR gave misleading results. CONCLUSION: Competitive RT-PCR is a valuable tool for molecular monitoring during postremission chemotherapy, as well as after BMT.

Gardner's syndrome in several family members.

This article describes chronologically a case of a fatal hereditary disease, which manifests itself in multiple polyps in the large intestine, accompanied by the appearance of numerous mesenchymal tumors on and under the skin and in the bones. In the Internal Department of the General Hospital Murska Sobota, we diagnosed the disease in one member of the family; in another, we ascertained the illness was retrograde. We confirmed the two cases in our dispensary; one of them remains under the constant surveillance of a gastroenterologist. The disease was in an advanced stage in the two family members at the time of discovery, so the treatment was radical. Nevertheless it terminated fatally. We expect to undertake future radical surgical measures on the other member in whom the disease was diagnosed. In this article, we illustrate the course of Gardner's Syndrome (GS), where in spite of early diagnoses, which now begin with molecular genetics and continues to endoscope examinations, and radical operative intervention, the quality of life of the patient is not improved to any great degree. The patient represents an exceptionally high risk group for the development of other mesenchymal tumors growths and colorectal cancer. The open question is also what is surgical treatment: total colectomy or proctocolectomy and what to do with the ileum when it is full of polyps.

[Numerical density of eosiophilic leukocytes in skin basal and squamous cell carcinomas]. / Brojna zastupljenost eozinofilinih leukocita u bazocelulnom i spinocelulnom karcinomu koze.

The autors examined the presence and determined numerical density of eosinophilic leukocytes in tumor's stormal and host reaction in twenty routine preparated skin biopsies of basal and squamos cell carcinomas. The presence of eosiophilic leukocytes in squamous cell carcinomas, in stormal and adjacent tumor infiltrate were confirmed, so as the absance on the same localizations in basal cell carcinomas were found. Eosinophilic leukocytes were of caracteristic morphological appereance, smaller of blood and bone marrow eosinophils - the average radius were 9,6 microm, with high level of degranulation, very variable in number (from 1054 to 23560 in mm3) and in close relation with plasma cells, mast cells and lymphocytes. In conclusion, these results may indicate and clear up presence of eosinophils by antigen properties of tumor cells, by activation of humoral immunity and consecutive phagogytosis of antigen-antibody complexes by eosinophils, but specially by their novel lytical effects on epithelial tumor cells in texture of cellular cytotoxic response. Beside these favorable effects of eosinophils, their granular enzymes activated and released during degranulation paradoxlly make spreading of tumor process possible and they reduce barrier role of dermal collagen.