Biosatellite 3. Preliminary findings.

Physiological deterioration in the male macaque monkey flown in Biosatellite III necessitated its recall after 8.5 days of a planned 30-day flight. For the first 7 days the only telemetered signs of a progressive general decline were falling brain temperature and lowered central venous pressure, which occurred in the last 3 days of flight. Fluid loss in flight was high, caused initially by sweating and later by diuresis, and appeared to arise in redistribution of blood in visceral pools as a consequence of weightlessness. Death occurred suddenly 12 hours after the flight and was caused by ventricular fibrillation.

Efficacy of antitumor chemotherapy in C3H mice enhanced by the antiangiogenesis steroid, cortisone acetate.

The effect of "antiangiogenesis" therapy using cortisone acetate (CA) with or without heparin on tumor growth as well as in combination with chemotherapy was investigated. C3H mice were implanted intradermally with N-[4-(5-nitro-2-furyl)-thiazolyl]formamide induced undifferentiated transitional cell carcinoma, MBT-2, in the right flank. The treatment was initiated 9 to 10 days after tumor inoculation. Daily injections of CA (250 mg/kg s.c.) suppressed tumor growth significantly in a dose dependent fashion. Administration of heparin (Elkins-Sinn) at the concentration of 200, 400, or 1000 units/ml in drinking water for 3 to 6 days was neither additive nor detrimental to the effect of CA. Chemotherapy was combined with CA; 3 days of administration of 250 mg/kg of CA in tapering doses was used. The chemotherapeutic agent was injected once 24 h before the initial CA. Combinations of chemotherapy (Adriamycin, 2.5-7.5 mg/kg i.v; cisplatin, 3-9 mg/kg i.p.; cyclophosphamide, 50-150 mg/kg i.p.; cis-(diammino)(1,1-cyclobutanedicarboxylate)platinum(II) (JM-8), 60-150 mg/kg i.p.; mitomycin C, 3-4.5 mg/kg i.p.) with CA showed additive suppression of tumor growth. Mice tolerated chemotherapy alone, CA alone, and both in combination. CA combined with JM-8 was not tolerated. Mice tolerated 100 to 150 mg/kg of JM-8, whereas the addition of CA to JM-8 resulted in a 66% (6 of 9) to 89% (8 of 9) mortality rate. CA at a concentration of 5 and 25 micrograms/ml showed no direct cytotoxic activity against MBT-2 cells in vitro. However, 3 days of administration of 250 mg/kg of CA inhibited tumor angiogenesis generated by MBT-2 cells in C3H mice using a dorsal air sac assay. The data suggest that CA alone inhibits tumor angiogenesis in C3H mice and that antiangiogenesis therapy enhances the antitumor efficacy of chemotherapeutic agents without increasing host toxicity (except for JM-8).

Immunohistochemical localization of parathyroid hormone-related protein in human prostate cancer.

Parathyroid hormone-related protein (PTHrP) is produced by a variety of malignant tumors and has been implicated as a major cause of humoral hypercalcemia of malignancy. Expression of PTHrP in prostate cancer tissue was studied immunohistochemically using 33 radical prostatectomy specimens from patients with clinically localized carcinoma of the prostate. None of these patients demonstrated hypercalcemia prior to the surgery. Acetone-methyl benzoate-xylene-processed, paraffin-embedded tissues were stained with a validated mouse monoclonal antibody to an amino acid fragment, PTHrP(109-141), using the streptavidin-peroxidase enzyme conjugate method. All cases (33 of 33; 100%) studied demonstrated some degree of immunoreactivity throughout the cytoplasm of the tumor cells, but immunostaining was absent from inflammatory and stromal cells. The intensity of the staining appeared to directly correlate with increasing tumor grade. The widespread immunohistochemical localization of PTHrP in carcinoma of the prostate suggests that PTHrP may play some local role in the growth of transformed cells in the prostate. Furthermore, overexpression of PTHrP may be a possible marker to evaluate the malignant potential of carcinoma of the prostate.

The androgen receptor status of neuroendocrine cells in human benign and malignant prostatic tissue.

Neuroendocrine (NE) cells containing neurosecretory granules, rich in various peptide hormones and biogenic amines, are components of the human prostate epithelium and prostatic adenocarcinomas. Neuroendocrine differentiation in prostatic adenocarcinomas has been associated with a poor prognosis and, following androgen withdrawal therapy, tumor cell populations have been observed to become enriched with NE cells. We assessed androgen receptor (AR) expression in NE cells in benign and malignant prostatic tissue using double-labeling immunocytochemistry with validated monoclonal antibodies to the AR and to chromogranin A (a generic NE marker). Neuroendocrine cells in benign and malignant prostatic tissue generally showed nuclear staining with AR. Some distinct AR-negative nuclei were observed in normal NE cells. In prostatic adenocarcinomas with extensive NE differentiation, a subpopulation of AR-negative NE cells was demonstrated. In conclusion, benign and malignant prostatic tissue contain both AR-positive and AR-negative NE cells that may have significance in regards to androgen-independent tumor growth and tumor progression.

Multiple molecular weight forms of immunoreactive alpha-inhibin in human seminal plasma.

The purpose of these studies was to determine whether the total immunoreactive alpha-inhibin protein concentration in seminal plasma correlated with serum gonadotropin levels or semen characteristics and to identify the forms of alpha-inhibin present in seminal plasma. Thirty-eight serum samples from men being evaluated for infertility were selected for study based on their serum hormone profiles and semen parameters. Serum LH and testosterone levels were normal, but FSH levels ranged from normal to hypergonadotropic (> 20 IU/L). Most semen parameters were within normal ranges, but germ cell numbers ranged from normal to azoospermic. Thus, seminal plasma from these men provided a unique opportunity to examine the antigenic forms of alpha-inhibin in individuals in whom strong correlations between inhibin and FSH levels might be predicted because of the observed ranges of FSH levels and germ cell numbers. Seminal plasma alpha-inhibin was characterized by RIA or Western blotting, using an antiserum directed against the N-terminal of the alpha-subunit of mature [32,000 mol wt (M(r))] inhibin. The antiserum recognized the alpha-subunit of dimeric inhibin as well as free alpha-inhibin and alpha-inhibin precursor proteins. Total immunoreactive alpha-inhibin ranged from 8.21-43.99 nmol/L in seminal plasma. However, alpha-inhibin levels were not statistically correlated with serum FSH levels or any of the measured semen parameters (including germ cell number). In contrast, the immunoreactive alpha-inhibin concentration in seminal plasma was negatively correlated (P < 0.01) with the serum LH level. Western blot analyses revealed that multiple forms of immunoreactive alpha-inhibin are present in seminal plasma. The majority of immunoreactivity was associated with monomeric proteins (ranging from 58,000-95,000 M(r)) that were larger than the alpha-subunit (21,000 M(r)) predicted for mature dimeric human inhibin (32,000 M(r)). The relative amounts of individual forms of immunoreactive alpha-inhibin varied among the patients studied, but could not be correlated with other serum or seminal parameters measured. Our observations demonstrate that various monomeric alpha-inhibin proteins are present in human seminal plasma. It is unlikely that these proteins alone or combined with inhibin beta-subunit proteins have identical biological activities. Thus, until assays specific for each of the various forms of immunoreactive alpha-inhibin are developed, their role as well as that of inhibin in the endocrine or local modulation of testicular function cannot be deduced from RIA data alone.

Detection of sperm surface IgG by modified haemadsorption procedures.

New haemadsorption procedures for the detection of sperm bound IgG are described. Antibody-coated sperm are fixed with formaldehyde and attached by poly-L-lysine to the bases of wells on plastic plates where they can be left for several days if necessary. Sensitised indicator red cell (for direct haemadsorption) or anti-IgG sera followed by indicator red cells (for indirect haemadsorption) are then added to these wells and the presence of antibody indicated by red cell attachment to sperm. The procedure uses simple reagents and the sperm/red cell rosettes may be fixed and stained to be kept as records.

Successful thrombolytic therapy in two patients with renal vein thrombosis.

Thrombolytic therapy was administered to two patients with angiographically documented renal vein thrombosis. One patient with bilateral thrombotic disease complicating membranous glomerulonephritis was treated with systemic streptokinase; the second patient had unilateral renal vein thrombosis associated with chronic pyelonephritis and was treated with streptokinase and urokinase administered directly into the involved vessel. Lysis of all thrombus material was demonstrated by follow-up renal venography and was associated with a gratifying improvement of clinical well-being and laboratory function. These cases demonstrate the feasibility of dissolving renal vein thrombi of presumed recent onset and suggest that rapid lysis of the thrombus accelerates functional and clinical recovery.

Androgen receptor immunostaining and its tissue distribution in formalin-fixed, paraffin-embedded sections after microwave treatment.

We used microwave (MW) oven heat treatment to unmask human androgen receptor (AR) immunostaining in formalin-fixed, paraffin-embedded tissue. Prostate tissue was used as an AR-positive control. Tissue sections were boiled in citrate buffer in a conventional MW oven for 30 min, followed by immunostaining with a validated murine monoclonal antibody (MAb), F39.4.1, raised against a peptide included in the N-terminal domain of the 100 KD human AR. AR immunostaining was localized to the nuclei of prostate secretory epithelial cells but was weak or absent in basal cells and of variable intensity in the stromal cells. Slides exposed to less than 10 min of MW heat treatment or none at all manifested no AR immunoreactivity. Tissue morphology was well preserved. Immunohistochemical determination of AR status in a wide variety of human tissues was consistent with that previously reported by others using frozen sections. MW heat treatment of tissue samples in an excellent method of localizing AR antigenicity, enabling immunohistochemical evaluation of AR status in formalin-fixed, paraffin-embedded material.

Differential effects of cyclosporin on hepatic and renal heme, cytochrome P-450 and drug metabolism. Possible role in nephrotoxicity of the drug.

Treatment of rats with 25 or 50 mg/kg cyclosporin A for 6 days elicited vastly different responses in hepatic and renal heme and drug metabolism activities. In the liver, cytochrome P-450 concentration was decreased significantly (to 70-75% of the control). This was accompanied by a marked reduction in benzo[a]pyrene hydroxylase activity (to 20-28% of the control). Aniline hydroxylation was also decreased, but to a lesser extent (to 77% of the control). In contrast, in the kidney cytochrome P-450 concentration was significantly increased to (145-170% of the control), along with a modest decrease in benzo[a]pyrene hydroxylation activity. In this organ, the concentration of porphyrins was severely decreased (to 30% of the control). Also, the activities of delta-aminolevulinate (ALA) synthetase and ALA dehydratase, as well as that of heme oxygenase, were inhibited. It is suggested that in the kidney the inhibition of degradation, rather than an enhanced rate of synthesis of the heme molecule, contributes to the observed increase in cytochrome P-450 concentration. In the liver, the decrease in the cytochrome concentration could not be explained in terms of an alteration in the rate of heme biosynthesis or degradation. Therefore, the observed decrease in cytochrome P-450 concentration could reflect the direct inactivation of the hemoprotein or regulation of apoprotein production by cyclosporin and/or its metabolite(s). The possible relevance of the observations to cyclosporin nephrotoxicity is discussed.

Immunohistochemical localization of parathyroid hormone-related protein in prostatic intraepithelial neoplasia.

Parathyroid hormone-related protein (PTHrP) is a regulatory protein hormone that has been associated with normal fetal growth and differentiation as well as fetal calcium regulation. Parathyroid hormone-related protein has been implicated in a variety of carcinomas as a major factor in the development of humoral hypercalcemia of malignancy and may also play a role as an autocrine growth factor. In a previous immunohistochemical study we found that all prostatic adenocarcinomas (CAP) express PTHrP. In the current study, we evaluated PTHrP in prostate intraepithelial neoplasia (PIN) in radical prostatectomy specimens. A validated mouse monoclonal antibody, 9H7, raised against fragment 109-141 of the carboxy-terminus of PTHrP was used for immunostaining. The results generally showed negative to weak staining of normal and hyperplastic tissue and strong staining in PIN. The staining intensity was further evaluated by computer based image analysis. The relative optical density in PIN (9.24 +/- 9.05) was significantly (P = .008) higher than that in normal gland (.00 +/- 3.6). These findings suggest that PTHrP may be involved in the pathogenesis of prostatic dysplasia, and its immunohistochemical evaluation may have diagnostic use in the evaluation of PIN.

Improved use of buthionine sulfoximine to prevent cisplatin nephrotoxicity in rats.

Male Sprague Dawley rats were treated with buthionine sulfoximine (BSO) and cisplatin in different doses and schedules to optimize the chemoprotective effect of BSO against cisplatin nephrotoxicity. BSO at 4 mmol/kg, administered s.c. 2 h prior to cisplatin, resulted in normal blood urea nitrogen (BUN) levels in rats treated with 3 or 4 mg/kg cisplatin, and modestly, but significantly reduced the toxicity of cisplatin at 5 mg/kg. Administration of BSO (4 mmol/kg) at intervals ranging from 0 to 16 h prior to cisplatin (5 mg/kg) resulted in a significant reduction in BUN values. A BSO dose as low as 0.04 mmol/kg was found to be as effective as 4 mmol/kg against nephrotoxicity associated with cisplatin at 4 mg/kg. Repetitive injections of BSO (1 mmol/kg every 12 h, four times, beginning 2 h prior to cisplatin) significantly inhibited elevations of BUN associated with higher-dose cisplatin (6 mg/kg), whereas a single BSO injection of 4 mmol/kg was ineffective. The degree and duration of renal glutathione depletion was related to the dose of BSO. Renal glutathione content following 4 mmol/kg BSO was 38% of control at 2 h and 40% at 24 h; following 0.04 mg/kg, glutathione was 47% at 2 h and almost 100% at 24 h. Simultaneous in vitro administration of BSO did not inactivate cisplatin cytotoxicity as measured by the colony-forming ability of MBT-2 cells in soft agar. These data indicate that repeated injections of BSO, beginning prior to cisplatin administration, would improve the nephroprotective effect without compromising the chemotherapeutic efficacy of cisplatin. It is suggested that the ability of BSO to reduce cisplatin nephrotoxicity may not correlate with the degree of renal glutathione depletion and that the mechanism of action is unlikely to involve direct inactivation of cisplatin.

Pathophysiology of bends and decompression sickness. An overview with emphasis on treatment.

Current concepts in the pathophysiology of decompression sickness are reviewed. Mild, moderate, and severe forms of this syndrome resulting from gaseous and lipid emboli are described. Therapy is aimed at restoring or specifically treating each alteration. Plasma volume deficit is restored by colloidal re-expansion. Decompression sickness is partially treated when recompression alone is used. Blood lipid alterations are managed by use of antilipemic agents. Dextran is mentioned. Divers at depths of 61 m display changes in hematocrit, platelet, and blood lipid profiles. Cord paralysis may occur from bubbles in the vena cava. Retrograde migration blocks the venous circulation of the spinal cord. Ultrasonic devices can detect "silent" bubbles during decompression. Recompression, when available, is a lifesaving treatment for diving accidents involving saturation diving. Air embolism is discussed. Monitoring emboli by EEG and fundoscopy are reported.

Lymphatic network of kidney. I. Anatomic and physiologic considerations.

A small number of investigators, including our team, has been responsible during the past two decades for many studies focusing on the kidney lymphatics. Interesting new information and concepts have been published, but unfortunately some of these data are scattered in obscure journals. The major purpose of this, and three subsequent reviews, is to present these new concepts in a more organized fashion. The reviews will also allow our group an opportunity to emphasize physiologic principles which have been ignored in the past. In a future review we will emphasize the role of the immunoblast, found only in kidney lymph and responsible for rejecting the renal allograft. Azathioprine (Imuran), if given early and in adequate dosage, will reduce the role of these transformed lymphocytes. Still another review article will emphasize the importance of renal tissue concentrations of antibacteria in eradicating acute pyelonephritis. Blood and urine levels of antibiotics are also important in eradicating pyelonephritis. Since renal lymph represents interstitial fluid, the concentration of appropriate bacteria-sensitive antibacterials is probably more important. We have now evaluated fourteen antibacterials individually in the mongrel dog, comparing plasma, urine, and renal lymph levels during similar time intervals. Finally, we will review the role of diuretics in altering the interstitial gradient of various solutes. Osmotic diuretics of more potent diuretics which affect renal tubular reabsorption manifest their action by different means when renal lymph fluid is evaluated.

Marcello Mapighi--a tribute.

Marcello Malpighi was probably the best known member of the faculty at the Medical School of Bologna. He was an anatomist, physiologist, botanist , and an entomologist . It was not until his appearance that the real value of the microscope was appreciated. He studied the embryologic form of the kidney and depicted in drawings how it was modified into the adult. He demonstrated the capillary anastomosis between the arteries and veins which was implicit in William Harvey's writings. He was a physician to Pope Innocent XII. Most historians call him one of the most charming personalities in the history of medicine.

Infertility and genitourinary infection.

There is a vast amount of evidence linking the presence of genitourinary infection with infertility. Infection in either partner can result in specific effects on sperm function, accessory sexual gland dysfunction, and the induction of immunologic responses in either partner. Careful investigation for infection should be undertaken in every couple with the complaint of infertility. Treatment needs to be individualized in each instance.

Topical levarterenol in control of bladder hemorrhage.

Topical levarterenol was evaluated for the control of bladder hemorrhage. Its possible toxic effects were investigated in the intact dog bladder, and its use was compared with saline irrigations in experimental massive bladder hemorrhage. Eight mg.% levarterenol was found to be safe and effective in our model.

Computed tomography of bladder: staging of bladder cancer using low density opacification technique.

In the course of computed tomographic (CT) evaluations involving 200 patients with suspected pelvic disease we have found that scan quality is often suboptimal. A preliminary report employing a low density bladder opacification method is presented whereby an indwelling catheter is inserted to control bladder volume and facilitate the instillation of low density iodinated contrast agent (0.6% Renografin-60). The patient remains supine throughout the twenty-five-minute procedure. Observations during the CT staging and follow-up of a controlled group of 8 patients undergoing immunotherapy and/or radiation therapy for bladder carcinoma indicate that this is a simple, safe, and effective staging procedure. We have obtained reliable clinical-radiologic-pathologic corrlation as a result of careful surgical staging and biopsy, followed by open surgery and full pathologic examination. It is hoped that this information will help initiate other similar studies to determine the diagnostic accuracy of this method and thus its use in pretherapeutic evaluation of the bladder lesion and its subsequent response to treatment.

Relationship between testicular volume and presence of varicocele. A comparative study.

The varicocele, present in many of the male partners of infertile couples continues to generate controversy, particularly as related to its diagnosis and pathophysiology. The purpose of our study was to determine the relationship between testicular volume and the presence or absence of a varicocele. Testicular volume was determined by the use of an orchiometer; the presence of a varicocele was determined by palpation during a Valsalva maneuver. The patients utilized in the study were those seen in our Andrology and General Urology Clinics; 291 patients with varicoceles and 83 control patients (no evidence of varicocele) were used. The left (mean = 21.4 mL) and right (mean = 23.4 mL) testicular volumes of patients with a varicocele were significantly reduced compared with that of the control group (left, mean = 23.4 mL; right, mean = 26.2 mL; p = 0.0041 and p = 0.002, respectively). The testicular volume corrected on the basis of the body mass (V/m2) also was significantly reduced in the varicocele group compared with controls (left p = 0.007; right p = 0.001). Reduced testicular volume relative to body size may be detected prior to actual demonstration of the stress pattern and the presence of a varicocele, and may be useful in early diagnosis. It is suggested that measurement of testicular volume may be a useful adjunct to routine examination of the infertile male.

Zinc sulfate therapy for infertile male with or without varicocelectomy.

One hundred one men were treated for infertility with 440 mg of zinc sulfate daily for sixty days to two years. All subjects demonstrated low seminal plasma zinc concentrations prior to treatment. Patients were divided into two groups: group [Zn], 65 subjects who underwent zinc therapy alone and group [Var/Zn], 36 subjects who received zinc sulfate therapy after varicocelectomy. In both groups seminal zinc levels were significantly higher (P less than 0.05 and P less than 0.01, respectively) two months after the initiation of zinc therapy. There were no significant changes in sperm count or motility for the [Zn] patients. However, group [Var/Zn] showed a significant increase (P less than 0.05) of sperm motility at two months and twelve months after starting zinc sulfate therapy. We separated the [Var/Zn] patients into two sub-divisions: those who impregnated their wives after therapy, and those who remained infertile despite treatment. Only those patients who impregnated their wives showed a significant (P less than 0.05) increase in sperm motility two months after the initiation of zinc therapy. Those patients of the [Var/Zn] group who remained infertile despite treatment showed a significant increase (P less than 0.05) in sperm motility after twelve months of therapy. Of the [Zn] patients 27.7 per cent (18) successfully impregnated their wives, while the [Var/Zn] patients exhibited a pregnancy rate of 50 per cent (18 patients). These data indicate that zinc sulfate therapy for patients with low seminal zinc concentrations may be an effective treatment for infertile patients especially after varicocelectomy.

Effect of low-dose androgen and zinc sulfate on sperm motility and seminal zinc levels in infertile men.

Beneficial effects of zinc therapy in some infertile patients with low seminal zinc concentrations and decreased sperm motility have been reported recently. The effects of zinc therapy, low-dose (5-10 mg/day) androgen therapy, and combination therapy of both zinc and low-dose androgen, on sperm motility and seminal zinc concentration were studied in patients referred to the andrology clinic. Low-dose androgen therapy increased seminal zinc concentrations and sperm motility, only when pretreatment seminal zinc concentrations were low. Furthermore, patients with low seminal zinc concentrations and poor sperm motility showed greater improvement of sperm motility and seminal zinc levels in response to zinc sulfate with fluoxymesterone than did fluoxymesterone alone or zinc sulfate alone, possible due to a synergistic effect of zinc and androgen.