Hyperplasia and carcinoma in secretory endometrium: a diagnostic challenge.

The diagnosis of endometrial hyperplasia or carcinoma in a background of secretory endometrium can be difficult. We attempt to establish the diagnostic criteria to be used in such cases. We examined 80 cases of endometrial hyperplasia, carcinoma, and other conditions with glandular crowding arising in secretory endometrium, analyzed their morphologic features, assessed the volume percentage stroma in each case and performed Ki67 immunostaining on 27 cases. Thirteen cases each of secretory and gestational endometrium served as controls. The mean age of the patients was 45 yr. The non-neoplastic diseases included simple hyperplasia without atypia (56%), endometrial polyps (12.5%), and chronic endometritis with glandular crowding (3%). The proportion of cases with complex hyperplasia without atypia was 10%. Neoplastic diseases included atypical complex hyperplasia (12.5%) and endometrioid carcinoma (6%). The secretory changes were usually less advanced in the hyperplastic glands than in the background endometrium. The morphologic features that best distinguished hyperplasia or carcinoma from secretory endometrium included glandular crowding that stood out from the background; architectural disorder (the long axes of the glands pointing in different directions or parallel to the endometrial surface); dilated, irregularly shaped glands, including budding or branching glands and staghorn-shaped glands; stroma of a polyp; cribriform or confluent glands in cases of carcinoma; nuclear atypia in cases of atypical hyperplasia and carcinoma; and crowded nonsecretory glands. The volume percentage stroma of neoplastic lesions was less than that of non-neoplastic ones (34% vs. 61%, P=0.000001) and that of secretory endometrium (34% vs. 68%, P=0.000038). Non-neoplastic lesions did not have significantly more crowded glands than secretory endometrium (61% vs. 68%, P=0.11). Gestational endometrium had more crowded glands than non-neoplastic lesions (39% vs. 61%, P=0.000004), an approximately equal volume percentage stroma with complex hyperplasia without atypia (39% vs. 43%, P=0.51), and less crowded glands than neoplastic lesions (39% vs. 34%, P=0.03). The Ki67 index of the neoplastic lesions was higher than that of the controls, including secretory and gestational endometria (positive nuclei per 100 epithelial cells, 44.8 vs. 4.6, P=0.0004), of the non-neoplastic lesions (44.8 vs. 5.4, P=0.002) and of complex hyperplasia without atypia (44.8 vs. 9.3, P=0.007). Hyperplasia and carcinoma in secretory endometrium can be diagnosed on the basis of increased glandular crowding, architectural irregularity, nuclear atypia, and increased Ki67 index.

Oral neurovascular hamartoma: a lesion searching for a name.

BACKGROUND: Neurovascular hamartoma (NVH), in particular in the oral cavity, is rarely described in the literature. The low number of cases may reflect a genuine rarity of the lesion, or it may be due to its being unrecognized and/or under-reported. OBJECTIVES: To investigate clinical and microscopic features of oral NVH and to define microscopic diagnostic criteria with emphasis on the differential diagnosis. METHODS: Archival cases diagnosed as oral NVH between 1999 and 2011 were retrieved; clinical and demographic data were collected, and a paired morphometric analysis was conducted, with each case of NVH a case of fibrous hyperplasia (FH) from the same oral location. The nerve bundle and blood vessel density were quantified in five microscopic fields at ×100 magnification. RESULTS: The study group included 25 oral NVH, 11 men and 14 women, aged 6-76 years, (mean 44). The majority occurred in the tongue (54%), followed by the buccal mucosa and lower lip (17% each), clinically presenting as asymptomatic 0.25-2.5 cm exophytic masses. Microscopic characteristics included poorly circumscribed masses of closely packed nerve bundles and blood vessels in a loose matrix, containing minimal or no inflammation. The mean nerve bundle density was significantly higher in NVH (4.28 ± 1.26) in comparison with FH (0.27 ± 0.27), (P < 0.00001), and mean vessel density was significantly lower (5.98 ± 1.4 vs. 7.8 ± 1.9, respectively), (P < 0.0003). CONCLUSION: Oral NVH may not be as rare as previously considered. Morphometric analysis demonstrated that NVH presents a separate distinct entity.

Granulosa cell tumor of the broad ligament: report of a case with emphasis on the differential diagnosis with female adnexal tumor of probable Wolffian origin.

This is a report of a paratubal adult granulosa cell tumor (GCT) located within the right broad ligament in a 62-year-old woman. These are rare tumors with only 8 cases reported so far. Because of an overlap of topographic, morphologic, and immunohistochemical features, it is not always possible to differentiate between the broad ligament GCT and female adnexal tumor of probable Wolffian origin (FATWO). Although nuclear grooving is not an exclusive feature of GCT and can be seen in a variety of other neoplasms, in the context of the differential diagnosis between broad ligament GCT and FATWO, the presence of this feature may be very useful in establishing the diagnosis of broad ligament GCT.

Nontoxic effect of tacrolimus on normally perfused isolated liver and protective effect on hypoperfused liver.

BACKGROUND: Cyclosporine A has been shown to detrimentally affect post-transplantation vascular tone. Tacrolimus (FK506), an immunosuppressant whose mechanism of action is similar to that of cyclosporine A, is more potent in vitro and has a reportedly high level of safety. The effects of tacrolimus on hepatic vasculature and metabolism in isolated, dually perfused (through both the hepatic artery and porta) rat liver under normal conditions and again in association with a state of hypoperfusion followed by reperfusion, imitating the liver's clinical state during organ transplant, were investigated. MATERIAL/METHODS: Three groups were perfused normally with Krebs-Henseleit solution and three groups were hypoperfused (75% flow reduction) for two hours. Saline, tacrolimus 4 ng/ml, or tacrolimus 40 ng/ml was injected in three normally perfused and three hypoperfused liver groups to determine drug effects under normal conditions and in low-flow-reflow state. Non-radioactive microspheres were later administered to all livers via the artery to assess microcirculatory patency. RESULTS: Tacrolimus did not affect the normally perfused livers. Liver hypoperfusion without treatment (saline injection) caused wet-dry weight ratio increase, abnormal increases in hepatic artery pressure and resistance values, and non-physiologically low oxygen extraction during reperfusion. Hypoperfusion + tacrolimus 4 or 40 ng/ml yielded values closer to those of the normally perfused livers. Finally, more microspheres were trapped in the hypoperfused+saline-treated liver circulation than in the normally perfused or hypoperfused+tacrolimus 4 or 40 ng/ml livers. CONCLUSIONS: Tacrolimus appears to be nontoxic in the isolated liver and to mitigate microcirculatory derangement associated with low-flow-reflow states.

Role of poly(ADP-ribosyl)ation during human spermatogenesis.

OBJECTIVE: Genomic stability of cells is known to be linked to their poly(ADP-ribosyl)ation capacity. We aimed to demonstrate, for the first time, the patterns of poly(ADP-ribosyl)ation during human spermatogenesis. DESIGN: Retrospective case-control study. SETTING: Teaching hospital. PATIENT(S): Azoospermic men who underwent testicular biopsy for sperm recovery. INTERVENTION(S): Testicular biopsy evaluation by immunohistochemistry for the expression of poly(ADP-ribose) polymerase-1 (PARP-1) enzyme and of poly(ADP-ribose) (PAR) (an indicator for PARP activity.) MAIN OUTCOME MEASURE(S): The subcellular localization of both markers in testes with full spermatogenesis (obstructive azoospermia), spermatocyte maturation arrest, or Sertoli cell-only syndrome. RESULT(S): Expression of both markers was localized in germ cell nuclei in full spermatogenesis: PAR expression, indicating PARP activity, was exhibited in round and elongating spermatids and in a subpopulation of primary spermatocytes. Strong immunoreactivity for PAR was identified in all of the spermatocytes in maturation arrest at the spermatocyte level. Sertoli cells lacked immunoreactivity for both markers, whereas other somatic testicular cells were rarely immunostained. CONCLUSION(S): The detection of PAR expression in germ-line cells and its subcellular localization in meiotic and postmeiotic prophases demonstrates chromatin modifications occurring during spermatogenesis and establishes a key role for poly(ADP-ribosyl)ation in germ cell differentiation, presumably to safeguard DNA integrity.

Cellular retinol binding protein-1 expression in endometrial hyperplasia and carcinoma: diagnostic and possible therapeutic implications.

Cellular retinol binding protein-1 (CRBP-1) contributes to the maintenance of the differentiative state of endometrial glandular cells through the regulation of bioavailability of retinol and derivatives, but its role in endometrial oncogenetic process remains unclear. Antibodies to CRBP-1, estrogen and progesterone receptors (ER and PR) were applied to paraffin sections of proliferative (n = 10) and secretory endometrium (n = 9), and to endometrial polyps (n = 6), simple (n = 7), complex (n = 3) and atypical endometrial hyperplasias (n = 9) as well as to 47 endometrioid carcinomas of different histological grade (G) (G1, n = 18; G2, n = 19; G3, n = 10). Four serous and two clear cell carcinomas were also examined. In glandular cells, CRBP-1 positivity was mainly cytoplasmic and rarely in the nuclei. CRBP-1 immunodetection was weakly positive in proliferative and low and focal in secretory endometrium and higher in atypical as compared to simple and complex hyperplasias. CRBP-1 expression in G1 endometrioid carcinomas was similar to that in atypical hyperplasias. In the latter, the highest CRBP-1 expression was observed in areas of squamous differentiation. Semiquantitative evaluation revealed a significant decrease of cytoplasmic CRBP-1 immunoreactivity with the increase of tumor grade. Among G3 endometrioid carcinomas, 60% were CRBP-1 negative, whereas the remaining cases showed a very low and focal positivity. Serous carcinomas were also CRBP-1 negative. When areas of different grading were present within the same tumor, less differentiated areas retained a lower CRBP-1 immunoreaction. The progressive decrease of CRBP-1 paralleled that of ER and PR immunodetection. RT-PCR in eight endometrioid carcinomas suggested a decrease of CRBP-1 with the increase of tumor grade also at transcriptional level. Our results indicate that CRBP-1 immunodetection may constitute an additional tool for histological grading of endometrial carcinoma. The CRBP-1 loss during the progression of endometrial cancer suggests a new potential target for pharmacological strategies aimed to counteract its progression by increased intracellular retinol bioavailability.

Inhibition of lymphokine-activated killer cells generation in vitro by soluble factors released from mixed human tumor and peripheral blood mononuclear adherent cells culture.

BACKGROUND: A variety of molecules produced by both tumor cells and normal cells reduce the activity of lymphokine-activated killer (LAK) cells. We tested the possible cross-regulation of mel-624 melanoma cells and adherent peripheral blood mononuclear cells (PBMCs) in affecting LAK cell activity. METHODS: PBMC adherent cells were cultured together with mel-624 melanoma cells. Supernatant was transferred to a 4-day LAK cells generation culture consisted of PBMC nonadherent cells and interleukin-2. LAK cytotoxic activity was tested in a 4-hour assay against Daudi tumor cells prelabeled with sodium (51)chromate. RESULTS: The supernatant produced within the first 48 hours of mixed mel-624 melanoma cell and adherent PBMC culture substantially (by 69 percent) reduced the generation of LAK cells, whereas the supernatant from either tumor culture or adherent PBMC culture had no effect. The inhibitory effect was manifested on the generation of LAK cells when autologous nonadherent cells were cultured with 1,000 units/ml IL-2, but there was no effect on mature LAK cell cytotoxic activity. Inhibition of LAK cell generation was partially dependent on protein synthesis and was not mediated by transforming growth factor ss (TGF-ss). CONCLUSION: Our results point toward the production of soluble, yet unidentified proteins, in mixed tumor-adherent PBMC cultures, which substantially reduced the induction of LAK cells in culture.

Uterine retiform sertoli-leydig cell tumor: report of a case providing additional evidence that uterine tumors resembling ovarian sex cord tumors have a histologic and immunohistochemical phenotype of genuine sex cord tumors.

We report a case of a retiform Sertoli-Leydig cell tumor of intermediate differentiation presenting as a uterine intracavity polypoid mass in a 63-year-old woman. In contrast to sertoliform endometrioid carcinoma and to hitherto reported uterine tumors resembling ovarian sex cord tumors (UTROSCTs), which are primarily characterized by tubular glands and solid tubules, this tumor, which most likely represents a UTROSCT, showed a large spectrum of histologic features typical of a genuine retiform Sertoli-Leydig cell tumor. The diagnosis was confirmed by a battery of immunohistochemical stains, which also served as a tool for differential diagnosis with other neoplasms. The tumor cells were positive for broad spectrum keratin (CK) CK18, vimentin, calretinin, and progesterone receptor. Only a few isolated cells stained for inhibin. The tumor cells were negative for CK7, CK5/6, epithelial membrane antigen (EMA), carcinoembryonic antigen (CEA), carbohydrate antigen 125 (CA125), thrombomodulin, 013 (CD99), melan A, alpha-fetoprotein (AFP), placental alkaline phosphatase (PLAP), alpha-1-antitrypsin, estrogen receptor, S100, neurone specific enolase (NSE), chromogranin, synaptophysin, desmin, caldesmon, and CD10. Divergent differentiation of uterine cells seems to be the most likely pathogenetic mechanism. To the best of our knowledge, no UTROSCT showing such a variety of histologic features indicative of a true sex cord tumor has been reported before.

Detection of calretinin expression in abnormal immature Sertoli cells in non-obstructive azoospermia.

The current study identified for the first time calretinin expression in abnormal Sertoli cells of azoospermic men who underwent testicular biopsy for sperm recovery and application of the retrieved sperm by in vitro fertilization techniques. Testicular biopsies with various spermatogenic impairments were evaluated immunohistochemically for the expression of the calretinin calcium-binding protein and the marker for immaturity of Sertoli cells, cytokeratin-18 (CK-18). Distribution of the markers was assessed in testes demonstrating a histological phenotype of mixed atrophy, Sertoli cell-only, or normal spermatogenesis (obstructive-azoospermia) and in men carrying a deletion in the azoospermia factor region located on the Y chromosome. Calretinin-immunopositive immature Sertoli cells revealed by co-localization of both markers, calretinin and CK-18, were identified in the mixed atrophy group in seminiferous tubules demonstrating spermatogenic failure. Sertoli cells expressing both markers were rarely detected in all other groups. Leydig cells in all the assessed biopsies expressed calretinin and served as a built-in control for immunoreactivity. This pattern of calretinin-selective expression in immature Sertoli cells suggests a functional relationship between calretinin expression and the degree of Sertoli cell differentiation. Disorders of Sertoli cell differentiation as indicated by calretinin and/or CK-18 expression contribute to the multifactorial mechanisms underlying spermatogenic failure.

Gastric mucosa in a bronchogenic cutaneous cyst in a child: case report and review of literature.

We report a case of cutaneous bronchogenic cyst, partially lined by gastric mucosa of antral type in a 9-year-old boy. This is, to the best of our knowledge, the first report of gastric mucosa in bronchogenic cutaneous cyst in the literature.

Metastases of breast carcinoma to the uterus. Report of two cases, one harboring a primary endometrioid carcinoma, with review of the literature.

BACKGROUND: Metastases to the uterus are rare, accounting for less than 10% of all cases of metastases to the female genital tract from extragenital cancers. The endometrium is even less frequently affected by metastases. Lobular carcinoma is the most common type of breast cancer that metastasizes to the uterus. CASES: Two cases of infiltrating lobular carcinoma of the breast metastatic to endometrium and myometrium, one of them harboring an endometrioid adenocarcinoma, are reported. Both patients were on tamoxifen therapy and presented with uterine bleeding. CONCLUSION: To the best of our knowledge, uterine carcinoma serving as recipient of metastatic carcinoma from the breast has not been previously documented. This possibility should be considered when an unusual bimorphic pattern appears in a tumor until proven otherwise.

Sclerosing angiomatoid nodular transformation (SANT): report of 25 cases of a distinctive benign splenic lesion.

Twenty-five cases of a morphologically distinctive vascular lesion of the spleen are described. The patients were 17 women and 8 men, ranging in age from 22 to 74 years (mean, 48.4 years; median, 56 years). The most common presentations were incidental finding of an asymptomatic splenic mass (13 patients), abdominal pain or discomfort (6 patients), and splenomegaly (4 patients). None of the patients had evidence of recurrent disease after splenectomy. The splenic lesion was solitary, measuring 3 to 17 cm, and sharply demarcated from the surrounding parenchyma. The cut surface revealed a mass of coalescing red-brown nodules embedded in a dense fibrous stroma. All cases showed a remarkably consistent multinodular appearance at low-power examination. The individual nodules had an angiomatoid appearance, in the sense that they were composed of slit-like, round or irregular-shaped vascular spaces lined by plump endothelial cells and interspersed by a population of spindly or ovoid cells. Some of the nodules (particularly the smaller ones) were surrounded by concentric rings of collagen fibers. Numerous red blood cells were present, as well as scattered inflammatory cells. Nuclear atypia was minimal, mitotic figures were extremely rare, and necrosis was consistently absent. The internodular stroma consisted of variably myxoid to dense fibrous tissue with scattered plump myofibroblasts, plasma cells, lymphocytes, and siderophages. Immunostaining revealed 3 distinct types of vessels in the angiomatoid nodules: CD34+/CD8-/CD31+ capillaries, CD34-/CD8+/CD31+ sinusoids, and CD34-/CD8-/CD31+ small veins, recapitulating the composition of the normal splenic red pulp. These features are therefore different from those of littoral cell angioma, conventional hemangioma, and hemangioendothelioma of the spleen. We interpret these angiomatoid nodules as altered red pulp tissue that had been entrapped by a nonneoplastic stromal proliferative process. The characteristic morphologic appearance, immunophenotype, and benign clinical course suggest that this is a distinctive nonneoplastic vascular lesion of the spleen that we propose to designate as sclerosing angiomatoid nodular transformation (SANT).

Spermatogonial proliferation patterns in men with azoospermia of different etiologies.

OBJECTIVE: To demonstrate the pattern(s) of spermatogonial proliferation in different spermatogenic disorders. DESIGN: Retrospective case-control study. Teaching hospital. PATIENT(S): Azoospermic men who underwent testicular biopsy for sperm recovery and preparation for intracytoplasmic sperm injection. INTERVENTION(S): Testicular biopsy evaluation by quantitative immunohistochemistry for proliferating cell nuclear antigen (PCNA). MAIN OUTCOME MEASURE(S): The expression of PCNA in spermatogonia as an index of proliferating activity in testes with focal spermatogenesis, spermatocyte maturation arrest, or normal spermatogenesis. RESULT(S): In biopsies with focal spermatogenesis (11 men), there was a statistically significant reduction of PCNA-labeled spermatogonia in seminiferous tubules showing spermatocyte arrest compared with the expression in adjacent tubules with advanced spermatogenic stage or in normal spermatogenesis (obstructive azoospermia, six men). However, PCNA expression in tubules of the group with complete maturation arrest (six men) was significantly higher compared with the same spermatogenic defect-spermatocyte arrest-within focal spermatogenesis biopsies. CONCLUSION(S): Different causes underlie the spermatogenic disorders reported in this study. In focal spermatogenesis, the disorder is associated with the presence of mitotic inactive spermatogonia. The detection of normal active spermatogonia in the spermatocyte arrest group indicates that the spermatogenic defect, which is accompanied by meiosis impairment, is not related to a malfunction of spermatogonial proliferation.

Phenotype and function of lymphocytes from the neonatal umbilical cord compared to paired maternal peripheral blood cells isolated during delivery.

In the present study, we analyzed the immunological characteristics of mononuclear cells (MNC) isolated from both neonatal umbilical cord blood (UCB) and maternal peripheral blood (MPB) during the delivery. The in vitro proliferative response of UCB T lymphocytes was significantly reduced compared to the maternal response to phytohemagglutinin A, pokeweed mitogen, and alloantigen stimulation, in correlation with the lower percentage of UCB than MPB lymphocytes, but not with that of B cells. The mean cytotoxic activity level of interleukin-2 (IL-2)-activated natural killer (NK) was higher in UCB than in MBP, whereas the percentage of CD56(+) NK cell count was similar. Our results show differences in the immune reactivity of T and B lymphocytes from neonate and adult isolated under similar physiological conditions.

Computed tomography-guided core needle biopsy for bone and soft tissue tumors.

BACKGROUND: Imaging-guided core needle biopsy is a well-established technique for the diagnosis of bone and soft tissue tumors and tumor-like lesions in specialized orthopedic oncology centers. OBJECTIVE: To present our results of computed tomography-guided core needle biopsy with assessment of the accuracy of the technique. METHODS: Between July 1998 and October 2000, 215 CT-guided core needle biopsies were performed and histologically examined in the Unit of Bone and Soft Tissue Pathology, Tel Aviv Sourasky Medical Center. There were 80 soft tissue and 135 bony lesions. All biopsies were performed by the same radiologist and the histologic examination by the same pathologist. To assess the accuracy of the procedure, we compared the diagnosis at biopsy with the diagnosis after definitive surgery (when available). RESULTS: Bone core needle biopsy (n = 135) showed malignancy in 89 cases (primary or recurrent bone sarcoma, lymphoma, myeloma, metastatic carcinoma or melanoma). There were 29 benign lesions. In 17 cases the result was inconclusive and an open incisional biopsy was performed. Of the 80 soft tissue biopsies, 35 were malignant (25 soft tissue sarcomas, 6 lymphomas, 4 metastatic carcinomas); 40 were benign (myositis ossificans, neurofibroma, desmoid tumor, schwannoma, hematoma and others), and 5 were inconclusive and followed by an open incisional biopsy. The core needle biopsy histologic diagnosis was compared with that of the definitive surgery and the diagnostic accuracy was 90%. Only three samples initially diagnosed as benign turned out to be malignant. No significant complications occurred during the procedures. CONCLUSIONS: CT-guided CNB of musculoskeletal lesions is a safe and effective procedure that assures sufficient and proper material for histologic examination. The accuracy of this method in our center was 90%. Tumor sampling is extremely important, especially in soft tissue sarcomas, and cores should be taken in different directions, including areas of necrosis. The processing is quick, especially for bone CNB, and diagnosis can be achieved within 24 hours. The material undergoes excellent fixation and the immunostains are reliable.

Double immunolabeling by the RBM and the PLAP markers for identifying intratubular (in situ) germ cell neoplasia of the testis.

Identification of intratubular germ cell neoplasia (carcinoma in situ, CIS) of the testis is a diagnostic challenge, and markers are sorely needed to assist in accurately identifying the lesion. RNA-binding motif (RBM) protein, encoded by the Y chromosome, is expressed exclusively and consistently in differentiated male germ cells, while it is absent in neoplastic germ cells. Another immunohistochemical marker, placental alkaline phosphatase (PLAP), is commonly used for the detection of undifferentiated germ cells. The current study demonstrates that simultaneous use of the immunohistochemical markers, RBM and PLAP, by double immunolabeling enhances the accuracy of diagnosing CIS, a preinvasive testicular neoplasm.

Lack of RBM expression as a marker for carcinoma in situ of prepubertal dysgenetic testis.

Individuals with various intersex states who carry Y-chromosome material bear a high risk of developing testicular neoplasia. In order to gain more insight into the pathogenesis of this neoplasia, the current study evaluates the differentiation of the seminiferous epithelium in 46,XY dysgenetic male pseudohermaphroditism. Immunohistochemical evaluation was performed using the germ cell-specific RNA-binding motif (RBM) protein (encoded by the Y-chromosome) to identify normal germ cells, whereas placental alkaline phosphatase (PLAP) was used to detect neoplastic germ cells. Differentiation of somatic Sertoli cells was assessed using cytokeratin-18 (CK-18) and anti-Müllerian hormone (AMH) as markers for immature Sertoli cells. Specimens were taken from surgically removed dysgenetic gonads of five children (46XY karyotype). Intratubular germ cell neoplasia (carcinoma in situ [CIS] of the testis) was detected in all of them. Normal germ cells revealed immunoreactivity for RBM, whereas the PLAP-positive neoplastic germ cells were negative for RBM expression. Sertoli cells revealed an immature phenotype indicated by AMH expression in their cytoplasm. The design of the current study is unique in its assessment of the state of germ cell differentiation in dysgenetic gonads by the use of the RBM protein, which was expressed only in normal germ cells but not in those of CIS. Testicular dysgenesis interrupted the normal differentiation of the germ line and had no effect on the immature phenotype of the prepubertal Sertoli cells. This points toward the germinal component of CIS as the precursor for the promotion of testis cancer.

Immunohistochemistry Evaluation of the Effect in Vivo of Tumor Necrosis Factor (TNF)-alpha on Blood Vessel Density in Murine Fibrosarcoma.

PURPOSE: Angiogenesis is essential for tumor growth and metastases, thus bestowing obvious importance upon methodologies which could enable its inhibition. MATERIALS: C57BL/6 female mice bearing a subcutaneous (s.c.) MCA205 fibrosarcoma were used. METHODS: Ten mice were divided equally into two groups. One group was injected intraperitoneally (i.p.) with 10 mug tumor necrosis factor-alpha (TNF-alpha and the other (controls) with Hanks balanced salt solution (HBSS). Tumor growth was monitored at least twice weekly. The number of endothelial cells in the blood microvessels was assessed by immunohistostaining on paraffin-embedded tumor tissue sections using vascular endothelial growth factor (VEGF) and Factor 8 antibodies. Expression of the p53 gene was similarly assessed by immunohistostaining. RESULTS: Injection of 10 mug TNF-alpha into the tumor-bearing mice reduced the number of endothelial cells in the blood microvessels by 46% on day 3 post-injection which was accompanied by an increase (by 37%) in the expression of p53 in these cells. It also inhibited tumor growth compared to the HBSS-injected group starting at 17 days post-cytokine injection. DISCUSSION: The antitumor in vivo effect exerted by TNF-alpha on established murine sarcoma s.c. tumors may be due to an earlier effect of the cytokine on the tumor's blood microvessels, probably through an apoptotic mechanism involving the p53 gene.

Localization of the germ cell-specific protein, hnRNP G-T, in testicular biopsies of azoospermic men.

The increasing interest in the application of in vitro fertilization techniques in human reproduction has led to a wide use of testicular biopsies to identify the presence of spermatogenic foci in testes of azoospermic men. Histopathologic evaluation of these testicular biopsies is required to determine the spermatogenic state with respect to fertility potential and to rule out preinvasive testicular lesions. Heterogeneous nuclear ribonucleoprotein G-T (hnRNP G-T) is a germ cell-specific protein expressed most prominently during meiosis. We studied the usefulness of hnRNP G-T antibody in the evaluation of these biopsies and reasoned that its germ cell-restricted expression pattern might provide a marker to improve accuracy of diagnosis. Testicular biopsies with various spermatogenic impairments were evaluated immunohistochemically for hnRNP G-T expression. In biopsies exhibiting normal spermatogenesis (obstructive azoospermia), hnRNP G-T was localized in meiotic pachytene spermatocytes and round spermatids. Immunostaining was barely detected when maturation was arrested at the spermatocyte level and not at all in cases of Sertoli cell-only syndrome. Biopsies with a mixed histologic phenotype and minute concentrations of spermatogenesis demonstrated strong immunostaining only in tubules with full spermatogenesis. This distribution pattern of hnRNP G-T enabled instant identification of spermatogenic foci. Thus, exploitation of the hnRNP G-T marker, which is expressed preferentially as meiosis proceeds, enhances sensitivity and accuracy of diagnosis in the histologic evaluation of testicular biopsies.