Kidney enlargement and multiple liver cyst formation implicate mutations in PKD1/2 in adult sporadic polycystic kidney disease.

Distinguishing autosomal-dominant polycystic kidney disease (ADPKD) from other inherited renal cystic diseases in patients with adult polycystic kidney disease and no family history is critical for correct treatment and appropriate genetic counseling. However, for patients with no family history, there are no definitive imaging findings that provide an unequivocal ADPKD diagnosis. We analyzed 53 adult polycystic kidney disease patients with no family history. Comprehensive genetic testing was performed using capture-based next-generation sequencing for 69 genes currently known to cause hereditary renal cystic diseases including ADPKD. Through our analysis, 32 patients had PKD1 or PKD2 mutations. Additionally, 3 patients with disease-causing mutations in NPHP4, PKHD1, and OFD1 were diagnosed with an inherited renal cystic disease other than ADPKD. In patients with PKD1 or PKD2 mutations, the prevalence of polycystic liver disease, defined as more than 20 liver cysts, was significantly higher (71.9% vs 33.3%, P = .006), total kidney volume was significantly increased (median, 1580.7 mL vs 791.0 mL, P = .027) and mean arterial pressure was significantly higher (median, 98 mm Hg vs 91 mm Hg, P = .012). The genetic screening approach and clinical features described here are potentially beneficial for optimal management of adult sporadic polycystic kidney disease patients.

A pilot study of autologous cancer cell vaccination and cellular immunotherapy using anti-CD3 stimulated lymphocytes in patients with recurrent grade III/IV astrocytoma.

The study objectives were to determine; (1) whether activated T cells could be generated from peripheral blood of patients immunized with their own cancer cells, (2) whether adoptive transfer of the activated T cells to patients had toxic effects and (3) whether the infused cells produced clinical responses. Study patients had recurrent, surgically accessible grade III/IV astrocytomas. The patients were tapered off steroids after total surgical resection and immunized with autologous cancer cells plus Bacillus, Calmette and Guerin (BCG). Peripheral blood mononuclear cells were activated with anti-CD3, expanded with interleukin-2 (IL-2) and reinfused to patients. The number of activated T cells that was given back to patients varied between 10(10) and 10(11). Side effects that were observed following immunization and adoptive cell transfer included mainly transient flu-like symptoms. One patient's tumor partially regressed, but there was no effect on survival. Two other patients' tumors regressed, and the patients are apparently disease-free more than 5 and 4 years later. The other six patients' tumors were apparently unaffected by the treatment. Patient age, tumor grade and CD4/CD8 composition of infused cells were positively correlated with clinical responses. Cellular immunotherapy is feasible and is associated with minimal toxicity. Additional appropriately controlled studies will be required to determine whether cellular immunotherapy could be used as a treatment for central nervous system malignancy. Additional studies also will be required to determine the underlying immunological mechanisms.

Profile of prostatic-specific antigen in prostatic carcinomas.

Prostatic carcinomas (PCs) may be divided into two distinct categories: latent PCs, found mostly at autopsy, and clinical cases, which present with signs and symptoms. These two categories correspond fairly well to histologic grading of PCs and immunoperoxidase staining for prostatic specific antigen (PSA). The objective of this study was to find quantitative and qualitative differences if any, of PSA in PCs, corresponding to Gleason's histologic grade. By radiometric assay of PSA in tissue cytosol, PCs especially those of high histologic grade, were found to have lower PSA concentrations than normal and glandular hyperplastic prostatic tissue. Western blotting of cytosol was performed to detect differences between immunoreactive PSA of PCs compared with noncancerous tissue using both polyclonal and monoclonal antibodies against PSA. Western blotting with anti-PSA revealed some different bands between cancerous and noncancerous cytosols. Western blotting of cancerous and noncancerous cytosols was also performed using anti-prostatic acid phosphatase and anti-beta micro-seminoprotein. Reduced PSA concentration and different immunoblotting pattern of PSA were found to be characteristic for PCs, especially in carcinomas with grades higher than 7, which usually present with more aggressive invasion and metastases.

Successful treatment of a malignant rat glioma with cytotoxic T lymphocytes.

Brain tumors are highly resistant to therapy. Their diffuse infiltrative nature and the relative inaccessibility of brain tissue to blood and lymph are barriers to surgical and cytotoxic treatments alike. The purpose of this study was to produce immune cells specifically reactive with an anaplastic rat glioma (RT2) and determine whether those cells could affect tumor progression in the brain. RT2-specific cytotoxic cells were prepared by priming rats in vivo with RT2 tumor cells and Corynebacterium parvum and stimulating the primed lymphocytes in vitro with irradiated RT2 tumor cells and interleukin-2 (IL-2). Cultured cells exhibited a high level of cytotoxicity against RT2, but not C6 (an allogeneic glioma), 3M2N (a syngeneic mammary tumor), or CSE (a syngeneic fibrosarcoma) tumor cells. To generate a model for therapy, rats were injected intracerebrally with RT2, generating progressing brain tumors, which killed untreated rats in approximately 2 weeks. To test the therapeutic potential of the effector cells, tumor-bearing rats were treated by intravenous injection of lymphocytes on Day 5 of tumor growth. Treated rats also received a 5-day course of systemic IL-2 beginning on Day 5. Treatment with IL-2 alone, RT2-primed spleen cells, or RT2-primed spleen cells stimulated in vitro with C6 did not affect rat survival. However, tumor-bearing rats treated with RT2-stimulated lymphocytes exhibited increased survival or were cured. Systemic IL-2 was an essential adjunct, because survival was not affected by treatment with effector cells alone. Therapy initiated on Day 8 of tumor progression lacked effect on survival.(ABSTRACT TRUNCATED AT 250 WORDS)

Identification of herpes simplex virus infection by immunoperoxidase and in situ hybridization methods.

Seven cases of visceral herpes simplex virus (HSV) infection were observed in five cases of hematopoietic disease and in one case each of a newborn baby and a pregnant woman. These seven cases were studied with an immunoperoxidase method and in situ hybridization. In HSV lesions of squamous epithelium, the immunoperoxidase method using rabbit anti-human HSV revealed positive staining, mainly in the nucleus but with some cytoplasmic staining. DNA in situ hybridization revealed stronger positive staining in the nucleus. In HSV hepatitis positive staining was seen in the nucleus and cytoplasm, both by immunoperoxidase and in situ hybridization methods. In the newborn baby, HSV lesions were observed in the brain only, with numerous positive astrocytes identified by the immunoperoxidase method and a few positive astrocyte nuclei by in situ hybridization. Cultured human fetal fibroblasts from the lung were infected with HSV. The immunoperoxidase method revealed diffuse positive staining in the nucleus and in the cytoplasm whereas in situ hybridization revealed fibrillar positive staining in the nucleus only. Thus, the immunoperoxidase method using rabbit anti-human HSV can detect the presence of HSV protein more sensitively than in situ hybridization, probably because of the greater quantity of HSV protein compared with HSV DNA in infected cells.

Idiopathic dilated cardiomyopathy--an evidence of abnormal lipid accumulation accumulation in myocardium.

A case of idiopathic dilated cardiomyopathy was studied histologically and by quantitative determination of tissue lipid concentration in order to document increased lipid accumulation. Four chambers of the heart were dilated with left ventricular dominance. Microscopically, there was a mixture of primarily fragmented myocytes with sarcoplasmic degeneration and minorly hypertrophic myocytes. There were numerous areas of moth-eaten appearance of myocardium in all of the sections, which corresponded to fatty degeneration, composed of numerous small lipid droplets between decreasing contractile elements. Ultrastructurally, there were widened Z-bands in degenerated myocardium, corresponding to light microscopically enlarged contraction bands. The extent of moth-eaten appearance was more extensive in the left than in the right ventricle, which was also verified by a three-fold increase of triglyceride concentration in the left ventricle compared with age-matched control hearts. The moth-eaten appearance was also observed in the auricles and atria with less immunohistochemical staining intensity for atrial natriuretic polypeptide, demonstrating that this fatty change involves all parts of the myocardium.

Identification of cytomegalovirus infection in acquired immunodeficiency syndrome.

Cytomegalovirus (CMV) infection was observed in 10 of 12 autopsy cases of acquired immunodeficiency syndrome (AIDS) and appears to be the commonest life-threatening viral infection in AIDS. In all 10 cases, adrenal glands were affected with CMV and adrenal medullary necrosis was present in 6 cases. Lungs were affected with CMV in 7 cases with disseminated infection and positive CMV culture. In situ hybridization of tissue sections with CMV-specific DNA provided positive staining for CMV in inclusions as well as other infected cells without obvious inclusions. Human diploid lung fibroblasts were infected with isolated CMV in culture, yielding positive CMV identification within 5 days by in situ hybridization before specific cytopathic changes appeared in the fibroblasts. The early and specific detection of CMV is possible by in situ hybridization with cultured fibroblasts.

Disseminated histoplasmosis in acquired immunodeficiency syndrome: light and electron microscopic observations.

Two young homosexual men with acquired immunodeficiency syndrome (AIDS) presented with five-lobe pneumonia and maculopapular rash, respectively, and were found to have disseminated histoplasmosis by examination of peripheral blood smears. Bone marrow smears from one patient revealed more numerous Histoplasma capsulatum organisms than peripheral blood smears did. Electron microscopy of peripheral-blood buffy coat demonstrated histoplasma organisms in monocytes and neutrophils as well as tubuloreticular structures in small lymphocytes. A search for Histoplasma capsulatum in peripheral blood smears from patients with AIDS is warranted, especially in endemic midwestern states.

Different ionic forms of estrogen receptor in rat uterus and human breast carcinoma.

Estrogen receptors from rat uterus and human breast carcinoma were analyzed by diethylaminoethyl-cellulose chromatography. Cytosols which had been incubated for short periods of time demonstrated a single discrete elution peak, indicating a single ionic form, while cytosols incubated for longer periods of time generated a second ionic form of receptor. Addition of cations to cytosols also promoted the rapid appearance of this second ionic form of receptor. Either leupeptin, a protease inhibitor, or sodium molybdate prevented the appearance of this second ionic form of estrogen receptor. The estrogen receptor from rat uterine cytosol incubated without leupeptin or molybdate had a smaller apparent molecular weight than did estrogen receptor from cytosols incubated with leupeptin or molybdate. Altogether, these experiments suggested that a cation-dependent protease present in the cytosols from both tissues was degrading the estrogen receptor to a second smaller species during extended incubation times.

Partial purification of Rho (D) antigen from Rh positive and negative erythrocytes.

A rapid method is described for partial purification of Rho(D) antigen from sodium deoxycholate-solubilized erythrocyte membranes by affinity chromatography on a column coupled with anti-Rho(D) IgG. The Rho(D) antigen is a low molecular weight membrane protein that comigrates with the lipid zone of erythrocytes during sodium dodecyl sulfate/polyacrylamide gel electrophoresis. Similar quantities of Rho(D) antigen were present in Rh positive and negative erythrocytes. However, in the latter erythrocytes the antigen may not be exposed to the external cell surface, explaining why these cells are not agglutinated by anti-Rho(D) antiserum. LW antigen was also present in the fraction containing Rho(D) antigen. The genetic implications of these findings are discussed.

Electrophoresis of human multiple myeloma and Waldenström's macroglobulinemia sera in sodium dodecyl sulfate polyacrylamide gels.

Sera from normal persons and patients with IgA, IgD, IgG, and IgM monoclonal gammopathies were electrophoresed in polyacrylamide gels containing sodium dodecyl sulfate. The gels were stained with Coomassie blue or were used for immunodiffusion. By this method IgG multiple myeloma and IgM Waldenström's macroglobulinemia sera were readily distinguished by electrophoresis alone, whereas IgA and IgD myeloma sera were distinguished by further immunodiffusion against anti-alpha-chain antibody and anti-delta-chain antibody.

Characterization of monosomes produced by aflatoxin B1.

A single injection of 1.5 mg aflatoxin B1 per kg body weight produced approx. 70% disaggregation of rat liver polysomes into monosomes within 18 h. Isolated monosomes dissociated into 40 S subunits during centrifugation in linear sucrose gradients containing 0.3 M KCI. The 4 S to 5 S molar RNA ratio of the monosomes was calculated to be 0.6, indicating 0.6 tRNA and/or aminoacyl tRNA molecule per ribosome; no peptidyl tRNA was present. These results suggest that a single injection of affatoxin B1 produces monosomes which resemble runoff ribosomes.