Metabolic conversion of retinol to retinoic acid mediates the biological responsiveness of human mammary epithelial cells to retinol.

The biological effects of vitamin A are mediated in part by retinoic acid (RA) modulation of gene transcription. In this study, we examined whether normal human mammary epithelial cells (HMECs) are biologically responsive to retinol (ROH), the metabolic precursor of RA. While both ROH and tRA resulted in time- and dose-dependent decreases in total cell number, tRA was markedly more potent. Metabolically, treatment of HMECs with physiological doses of ROH resulted in rapid uptake and subsequent production of both retinyl esters and tRA. Although a comparatively minor metabolite, tRA levels peaked at 6 h and remained above endogenous levels for up to 72 h in proportion to cellular ROH concentrations. In HMECs transfected with an RA-responsive luciferase reporter gene, treatment with 3 microM ROH resulted in an increase in luciferase activity to a level intermediate between that observed with 0.001 and 0.01 microM tRA. Citral, an RA-synthesis inhibitor, was also used to examine the biological activity of ROH. Compared to ROH alone, ROH plus citral treatment resulted in three-fold less tRA synthesis and a > 65% attentuation of RA-responsive reporter gene activity which persisted through 72 h. Citral also significantly attenuated the extent of ROH-mediated reductions in total HMEC number. Thus, treatment with physiological concentrations of ROH results in fewer total numbers of HMECs and this response is a consequence of cellular tRA synthesis which can induce RA-responsive gene expression.

Acid suppression by famotidine 20 mg twice daily or 40 mg twice daily in preventing relapse of endoscopic recurrence of erosive esophagitis.

Control of esophageal acid exposure is important in treating patients with gastroesophageal reflux disease (GERD). After complete healing of esophagitis, most patients will relapse within 6 months if left untreated. This multicenter, randomized, double-masked, placebo-controlled trial, conducted in the United States, examined whether two famotidine dosing regimens are effective in extending the time in remission for patients with moderate-to-severe erosive esophagitis. Of 172 patients enrolled, 31 received placebo, 69 received famotidine 20 mg twice daily (BID) , and 72 received famotidine 40 mg BID. Endoscopy was scheduled at baseline and at months 3 and 6. Patients assessed global heartburn and symptom relief at months 3 and 6 relative to the start of the study. Life table (Kaplan-Meier) relapse rates at 6 months were 22% (P < 0.001 vs placebo) for famotidine 20 mg BID, 11% (P < 0.001 vs placebo) for famotidine 40 mg BID, and 62% for placebo. Compared with placebo, patients in the famotidine groups were significantly less likely to note global symptomatic deterioration, as measured by the distribution of global assessment responses. The incidence of clinical and laboratory adverse experiences was similar among treatment groups. For maintenance treatment of GERD, famotidine 20 mg BID and 40 mg BID are more effective than placebo in extending the time in remission.

Postnatal diagnosis and outcome of urinary tract abnormalities detected by antenatal ultrasound.

OBJECTIVES: To determine the incidence of urinary tract abnormalities detected by antenatal ultrasound; the value of fetal renal measurements in predicting significant uropathy; and the nature and clinical outcome of fetal uropathy. DESIGN AND SETTING: A retrospective analysis of babies with urinary tract abnormalities detected before birth who were born at or referred to a teaching hospital. PATIENTS: One hundred and twenty-five babies born between June 1989 and November 1992. Sixty-nine babies were born at Westmead hospital and 56 were born elsewhere and referred to Westmead Hospital for investigation. RESULTS: The incidence of uropathy detected before birth among babies born at the teaching hospital was 5.1 per 1000 births. In 60% of these babies, significant abnormalities were confirmed after birth (3.1 per 1000 births). We found a significance of 71% and a specificity of 89% for fetal measurements in predicting significant uropathy. Among all 125 babies the most common abnormalities were pelviureteric junction (PUJ) anomalies (34%), minimal hydronephrosis (26%), ureteric reflux (18%) and multicystic kidney (7%). Pyeloplasty has been performed in 23 (46%) of the 52 kidneys with PUJ anomalies. Renal parenchymal abnormality was detected in four of 22 kidneys with ureteric reflux (18%). Of these four, all were exposed to Grade IV or V ureteric reflux but none had been infected. CONCLUSIONS: Antenatal ultrasound is a sensitive, though nonspecific, tool for the non-invasive identification of congenital abnormalities of the urinary tract, which are common and frequently asymptomatic after birth. Although ureteric reflux may not be predicted from renal measurements, the degree of fetal hydronephrosis indicates the likelihood of obstructive abnormality and, thus, the extent of postnatal investigation required.

Chronic testicular pain.

The cause of chronic testicular pain is often difficult to define. Careful examination of the inguino-scrotal region is useful. Chronic orchalgia is a diagnosis of exclusion for which a multidisciplinary approach may be rewarding.

Patterns of expression of metalloproteinases and their inhibitors in human malignant lymphomas.

Forty-two cases of malignant lymphomas were studied for the expression of matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) by Northern blot and in situ hybridization. The lymphomas were classified according to Working Formulation classification (25 high grade, 15 low grade, and 2 intermediate grade). The MMPs studied included: 72-kDa and 92-kDa gelatinases (type IV collagenases), interstitial collagenase, PUMP-1 (MMP-7), and stromelysins 1 and 3. TIMPs included TIMP-1 and TIMP-2. All but one case expressed TIMP-1, and TIMP-2 transcripts were detected in 35 cases. Among the MMPs, 92-kDa gelatinase was expressed most consistently (35 cases), whereas mRNA transcripts of 72-kDa gelatinase, interstitial collagenase, and PUMP-1 were detected in only a few cases. Stromelysins 1 and 3 mRNAs were not detected in any of the tumors studied. However, marked differences in the level of expression of certain MMPs and TIMPs were found among different grades of malignant lymphomas. The low grade tumors expressed low and relatively constant levels of TIMP-1, TIMP-2, and 92-kDa gelatinase transcripts, whereas high grade lymphomas displayed variable amounts of mRNAs for TIMPs and MMPs, with a trend toward elevated TIMP-1 and 92-kDa gelatinase mRNA levels. In situ hybridization localized TIMP-1 transcripts to stromal cells, while 92-kDa gelatinase transcripts were most abundant in "starry sky" macrophages and large lymphoma cells. Zymography showed that active 92-kDa gelatinase is present in tumor protein extracts and differences in the level of the enzymatic activity were seen between low and high grade lymphomas. Our data indicate that 92-kDa gelatinase and TIMP-1 expression by human malignant lymphomas may play an important role in controlling their biologic aggressiveness.

Differential regulation of TIMP-1 and TIMP-2 mRNA expression in normal and Ha-ras-transformed murine fibroblasts.

A cDNA containing the complete coding region of the murine tissue inhibitor of metalloproteinases-2 (TIMP-2) was isolated using reverse transcription-polymerase chain reaction amplification. The predicted murine TIMP-2 amino acid sequence shows 96% identity with human TIMP-2, but only 42% identity with murine TIMP-1. This high degree of evolutionary conservation between the human and mouse proteins suggests that TIMP-2 performs an essential biological function. The expression of the TIMP-1 and TIMP-2 mRNAs was examined in normal and ras-transformed murine fibroblasts. While TIMP-1 transcription was highly serum-inducible in normal murine C3H10T1/2 fibroblasts, TIMP-2 mRNA expression was largely constitutive. A series of ras-transformed derivatives of C3H10T1/2 fibroblasts showed great variability in TIMP-1 expression: some lines retained serum inducibility, others displayed constitutive expression at either high or low levels. In contrast, TIMP-2 expression was insensitive to transformation. Neither TIMP-1 nor TIMP-2 expression at the RNA level, or total TIMP activity in conditioned media could be correlated with the metastatic potential of the ras-transformed lines. Our data demonstrate that the mechanisms that regulate murine TIMP-1 and TIMP-2 expression are distinct arguing for different physiological roles for the two TIMPs.

Exposure to low levels of hydrogen sulfide elevates circulating glucose in maternal rats.

Although the lethal effect of hydrogen sulfide (H2S) has long been known, the results of exposure to low levels of H2S have not been well documented. Rat dams and pups were exposed to low levels of H2S (less than or equal to 75 ppm) from d 1 of gestation until d 21 postpartum and analyzed for changes in circulating enzymatic activity and metabolites. Blood glucose was significantly elevated in maternal blood on d 21 postpartum at all exposure levels. This increase in glucose was accompanied by a possible decrease in serum triglyceride in the pups and in the dams on d 21 postpartum. There was no evidence of alterations in serum alkaline phosphatase, lactate dehydrogenase, or serum glutamate oxaloacetate transaminase.

Growth and development in the rat during sub-chronic exposure to low levels of hydrogen sulfide.

The effects of low levels of hydrogen sulfide (H2S) on mammalian growth and development are unknown although it has long been postulated that H2S can inhibit critical developmental functions through the cleavage of disulfide bonds and chelation of essential metal ions. Gravid rat dams exposed to H2S (less than or equal to 75 PPM) from day 6 of gestation until day 21 postpartum (PP) demonstrated normal reproductive parameters until parturition. At parturition, however, delivery time was extended in a dose dependent manner with a maximum increase of 42% at 75 PPM. Maternal liver cholesterol content was elevated significantly on day 21 postpartum following exposure to 75 PPM H2S each day for 6 weeks. Pups which were exposed in utero and neonatally to day 21 postpartum developed with a subtle decrease in time of ear detachment and hair development and with no other observed change in growth and development through day 21 postpartum.

Hydrogen sulfide exposure alters the amino acid content in developing rat CNS.

Hydrogen sulfide is a widespread environmental pollutant that may produce severe effects on the developing nervous system. Putative amino acid neurotransmitter levels in the rat cerebrum and cerebellum were determined to evaluate the effects of exposure to hydrogen sulfide during perinatal development. The levels of aspartate, GABA, glutamate, glycine and taurine were quantitated using high-performance liquid chromatography. With the exception of glycine, all of the amino acids examined were affected by the treatment. On day 21 postnatal, which was the last day of the exposure, aspartate, glutamate and GABA in the cerebrum and aspartate and GABA in the cerebellum were significantly depressed. The observed alterations in the amino acid levels during this critical phase of development may have chronically affected the activity of the neurotransmitters, their receptor sensitivity or their individual target areas. The consequence of one or a combination of such alterations may lead to behavioral and structural abnormalities.

Inhibition of oxytocin-induced but not angiotensin-induced rat uterine contractions following exposure to sodium sulfide.

Low concentrations (0.15-15 microM) of sodium sulfide reversibly attenuated the contractile response of the isolated rat uterus to oxytocin without affecting angiotensin II responsiveness. These findings suggest that functionally important disulfide bonds in the rat uterine oxytocin receptor, but not the angiotensin receptor, are sensitive to hydrosulfide ion. Reduction of oxytocin receptors by hydrosulfide ion may be a mechanism by which low levels of H2S delay parturition in rats.

Penile necrosis secondary to an indwelling Foley catheter.

Penile necrosis is a rare entity but it may be brought about more readily if certain risk factors are present. Such factors are discussed in this paper and the reader is forewarned about this problem.

Characterization of hyaluronate binding proteins isolated from 3T3 and murine sarcoma virus transformed 3T3 cells.

A hyaluronic acid binding fraction was purified from the supernatant media of both 3T3 and murine sarcoma virus (MSV) transformed 3T3 cultures by hyaluronate and immunoaffinity chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis resolved the hyaluronate affinity-purified fraction into three major protein bands of estimated molecular weight (Mr,e) 70K, 66K, and 56K which contained hyaluronate binding activity and which were termed hyaluronate binding proteins (HABP). Hyaluronate affinity chromatography combined with immunoaffinity chromatography, using antibody directed against the larger HABP, allowed a 20-fold purification of HABP. Fractions isolated from 3T3 supernatant medium also contained additional binding molecules in the molecular weight range of 20K. This material was present in vanishingly small amounts and was not detected with a silver stain or with [35S]methionine label. The three protein species isolated by hyaluronate affinity chromatography (Mr,e 70K, 66K, and 56K) were related to one another since they shared antigenic determinants and exhibited similar pI values. In isocratic conditions, HABP occurred as aggregates of up to 580 kilodaltons. Their glycoprotein nature was indicated by their incorporation of 3H-sugars. Enzyme-linked immunoadsorbent assay showed they were antigenically distinct from other hyaluronate binding proteins such as fibronectin, cartilage link protein, and the hyaluronate binding region of chondroitin sulfate proteoglycan. The apparent dissociation constant of HABP for hyaluronate was approximately 10(-8) M, and kinetic analyses showed these binding interactions were complex and of a positive cooperative nature.(ABSTRACT TRUNCATED AT 250 WORDS)

Receptors for epidermal growth factor (urogastrone) and insulin in primary cultures of rat hepatocytes maintained in serum-free medium.

We have analyzed the receptors for epidermal growth factor (urogastrone) (EGF-URO) and insulin in primary cultures of adult rat hepatocytes maintained for up to 3 weeks on human placental cell matrix in serum-free defined medium. Cross-link labeling experiments revealed that the insulin receptor, partially damaged by the collagenase isolation procedure, was rapidly regenerated to yield an intact receptor. In contrast, cross-link labeling of the EGF-URO receptor revealed that, upon prolonged culture, there was a progressive disappearance of the high molecular mass (175 kilodaltons (kDa)) receptor form, and an appearance of low molecular mass receptor species (130 and 105 kDa). After 3 weeks of culture, the low molecular mass receptor forms accounted for all of the labeled EGF-URO receptor present in the cells. Measurements of EGF-URO binding indicated that the number of EGF-URO binding sites per cell (2.0 x 10(5) +/- 0.3 x 10(5)) did not change during the 3 weeks of culture. However, there was a decrease in EGF-URO binding affinity, reflected by an increase in the KD from 0.6 to 3.0 nM. At zero time and after 3 weeks in culture, Scatchard plots of the binding data were linear; at intermediate time points, the plots were curvilinear. Despite the changes in the EGF-URO receptor that occurred, cells were still responsive to EGF-URO in terms of the inhibition of acetate incorporation into lipid. The ED50 for EGF-URO (about 0.2 nM) was the same for short-term cultures (48 h) as for cells maintained in culture for 3 weeks.(ABSTRACT TRUNCATED AT 250 WORDS)

Pathophysiology of ureteropelvic junction obstruction: experimental and clinical observations.

We compared experimental ligature-induced ureteropelvic junction obstruction in the dog with naturally occurring ureteropelvic junction obstruction in children to determine if clinical behavior and difficulties in diagnosis could be related to different types or components of obstruction at the ureteropelvic junction. Measurements of flow rate out of the ligature-obstructed canine renal pelvis demonstrated a pressure-dependent pattern in which flow increased linearly in response to increasing pressures. In 5 human kidneys with intrinsic ureteropelvic junction obstruction a similar pressure-dependent pattern was demonstrated. This was in contrast to 6 human kidneys with extrinsic mechanical ureteropelvic junction obstruction in which a volume-dependent pressure flow pattern occurred, such that urinary flow rate did not keep pace with increases in pelvic pressure. In some cases flow actually decreased at high pressures because the ureteropelvic junction became self-obstructing as the pelvis enlarged. These findings indicate that the precise pathological anatomy of the ureteropelvic junction defines the pattern of flow across the obstruction. The 2 different types of obstruction, pressure-dependent and volume-dependent flow restrictions, which exist are important determinants of the clinical behavior of the obstructed kidney insofar as its potential for progressive hydronephrosis. They also help to explain why diagnostic tests for assessing obstruction in hydronephrosis are inaccurate at times.

Inhibition of thymidine incorporation in primary rat hepatocytes by porcine pancreatic polypeptide.

The concentration of pancreatic polypeptide (PP), a peptide released by meal ingestion, was suppressed in obese mice and in humans, and earlier studies have suggested a metabolic function of PP in adipocytes and liver. These observations have prompted the examination of the metabolic role of PP in rat hepatocytes. These studies have examined the role of porcine PP in the control of [3H]-thymidine incorporation in adult rat hepatocytes maintained in the presence of insulin, glucagon and epidermal growth factor (EGF). Upon long-term exposure of cultured hepatocytes to porcine PP, basal (insulin and glucagon-maintained cells) and EGF-stimulated [3H]-thymidine incorporation were inhibited. Basal incorporation was inhibited with an ED50 of 23 pM. Thus, the long-term PP function may be suppression of stimulated thymidine incorporation and cellular replication.

Production of somatomedin-like activity by human adult tumor-derived, transformed, and normal cell cultures and by cultured rat hepatocytes: effects of culture conditions and of epidermal growth factor (urogastrone).

We have measured the production of a basic-somatomedin-like activity (SLA) by a variety of human tumor-derived, transformed, and normal postnatal cell cultures; and we have compared the production of SLA by these cell types with the production of SLA by adult rat hepatocytes cultured in serum-free medium. Cells derived from a human epidermoid carcinoma (KB), a pancreatic carcinoma (Panc-1), a Simian virus 40 transformed adult human skin-derived cell line (SV40 fibroblasts), and a normal adult human skin-derived fibroblast line released SLA when cultured in a serum-free growth medium. No SLA was recovered from the culture medium of human choriocarcinoma-derived cells (BeWo) or of a human lymphoblastoid cell line (IM-9). The production of SLA by rat hepatocytes cultured in serum-free medium appeared to exceed the production of SLA by the other cell cultures. In cultures of KB cells, SV40 fibroblasts, and rat hepatocytes, the production of SLA depended on the frequency with which the growth medium was renewed; in general, the highest rates of SLA production were observed when the medium was renewed every 48-72 h. The presence of mouse epidermal growth factor (urogastrone) (EGF-URO) in the serum-free culture medium stimulated the production of SLA by KB cells and by rat hepatocytes, but did not increase SLA production by normal or by SV40-transformed human skin-derived fibroblasts. We conclude that tumor-derived cells are capable of producing somatomedin-like activity and that the production of SLA by such cells can be subject to controls (nutrient availability, EGF-URO stimulation) that regulate SLA production, either by normal adult tissues, like liver, or by a variety of normal embryonic tissues.

Vesicoureteral reflux in triplets.

During an investigation for urinary tract infections in 3-month-old triplets similar patterns of vesicoureteral reflux were noted. To our knowledge, this is the first report of vesicoureteral reflux in triplets. The patterns of inheritance in vesicoureteral reflux and the need for sibling evaluation are discussed.

Diuretic radionuclide localization of upper urinary tract obstruction.

In 59 children with proved upper urinary tract obstruction diuretic radionuclide ureteral scans provided an accurate assessment of ureteral dilatation sufficient to distinguish ureteropelvic from ureterovesical obstruction. As a result, this test may be used instead of more invasive studies, such as retrograde or antegrade pyelography, to determine the site of obstruction in many cases of hydronephrosis.