Regenerative stimulus increases hepatocyte tight junctional permeability. Experience with young animals, animals administered certain hepatotoxins and animals with two-thirds hepatectomy suggests that tight junctional permeability is increased in states characterized by architectural remodeling in the liver. In this work we correlate changes in tight junctional morphometry induced by two-thirds hepatectomy with changes in biliary permeability assessed by sucrose and horseradish peroxidase permeation and by alterations in biliary outputs of anionic and cationic cholephilic probes. By freeze-fracture examination tight junctional strand counts, density and orientation parallel to canaliculi were all reduced 24 hr after two-thirds hepatectomy. Occasionally, strands were perpendicular to the canaliculi, creating an unobstructed communication between bile and intercellular spaces. These morphological changes correlated with increased sucrose and paracellular horseradish peroxidase access into bile, with reduced biliary outputs of low molecular weight and especially with cationic cholephilic probes. The data support an increased but still charge-selective permeability of the biliary tree, which was induced by two-thirds hepatectomy 24 hr before. Presumably, fixed intercellular connections (tight junctions and gap junctions) must be loosened or lysed to allow the architectural reorganization required by the hepatocellular regenerative process.


A pilot study of 2',3'-dideoxyinosine for the treatment of chronic hepatitis B. The nucleoside analog 2',3'-dideoxyinosine, currently being used to treat patients infected with the human immunodeficiency virus, has been shown to inhibit viral replication in certain cell culture systems of hepatitis B virus and the duck model of chronic hepatitis B infection. We studied the effect of dideoxyinosine on viral replication in patients with chronic hepatitis B. In the initial dose-finding phase, patients received sequential 2-wk courses of dideoxyinosine in escalating doses of 3, 6 and 9 mg/kg/day. In the second, long-term treatment phase, patients received dideoxyinosine at a dose of 9 mg/kg/day for 12 wk. Dideoxyinosine was given orally in three divided doses. The effects of dideoxyinosine on hepatitis B were assessed by serial measurements of ALT, hepatitis B virus DNA and DNA polymerase activity in serum. Six patients completed the dose-finding phase, and five patients continued into the long-term treatment phase. No significant differences were seen in serum aminotransferases, hepatitis B virus DNA levels or DNA polymerase activity at any time during treatment when compared with pretreatment levels. All patients remained positive for HBeAg during treatment and during 6 mo of follow-up. Thus at the doses tested, dideoxyinosine had no appreciable effect on viral replication in patients with chronic hepatitis B.


Characterization of the liver cytosol antigen type 1 reacting with autoantibodies in chronic active hepatitis. An autoantibody to liver cytosol was previously described in childhood autoimmune chronic active hepatitis type 2. The antigen, liver cytosol antigen type 1, was for the first time partially purified using gel filtration and ion exchange chromatography, and it was characterized using immunodiffusion, immunoblot and sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of the immunoprecipitate. Immunoblot detected a unique antigenic peptide at 62 kD from human cytosol and at 58 kD from rat cytosol. The same peptides were also detected when immunoprecipitates of liver cytosol antigen type 1 and autoantibodies to liver cytosol antigen were submitted to sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A polymeric structure, probably a tetramer, is suggested for native liver cytosol antigen type 1 because in gel filtration chromatography liver cytosol antigen type 1 was eluted as a protein of a molecular weight between 240 and 290 kD when human liver cytosol was fractionated and between 220 and 270 kD from rat liver cytosol. Liver cytosol antigen type 1 is probably poor in carbohydrates because it was not stained by periodic acid-Schiff stain. The autoantibodies to liver cytosol were frequently found in association with antiliver kidney microsomal autoantibodies type 1, which are directed against the cytochrome P-450 of the IID6 subfamily. Antiliver kidney microsomal autoantibodies type 1 but not antiliver cytosol autoantibodies were found in association with antibodies to hepatitis C virus. Autoantibodies to liver cytosol antigen type 1 seem to be a more specific marker for autoimmune hepatitis type 2 than antiliver kidney microsomal antibodies type 1 autoantibodies.


Antimitochondrial antibodies in kindreds of patients with primary biliary cirrhosis: antimitochondrial antibodies are unique to clinical disease and are absent in asymptomatic family members. The 2-oxo-acid dehydrogenase family of enzymes have been identified as the major mitochondrial autoantigens of primary biliary cirrhosis. Using immunoblotting, enzyme-linked immunosorbent assay and enzyme inhibition with both purified mitochondrial proteins and recombinant autoantigens, we have studied family members and spouses of patients with primary biliary cirrhosis for the presence of antimitochondrial antibodies. Antimitochondrial antibodies and other common autoantigens were also tested for by indirect immunofluorescence. This study included 27 index patients with primary biliary cirrhosis, 15 spouses and 48 first- and second-degree relatives. Overall, 7 relatives (11%) were positive for autoantibodies to nuclear and cytoplasmic antigens by indirect immunofluorescence against mouse liver and stomach sections. However, with immunofluorescence, the reactivity strictly paralleled that of antimitochondrial antibodies in only one of these (1:640)--a sibling with mild pruritus and a liver biopsy specimen diagnostic of primary biliary cirrhosis despite normal levels of serum alkaline phosphatase. In addition, one of the mothers, who had a history of sarcoidosis, was positive by immunoblotting for antibodies to the E2 subunit of the pyruvate dehydrogenase complex and protein X. All other relatives were negative for all of the assays. Antibodies to neither the 2-oxo-acid dehydrogenase enzymes nor the recently proposed family of naturally occurring mitochondrial antibodies were found in spouses or healthy relatives. Three other first-degree relatives suffered from liver disease: two died (one from primary biliary cirrhosis and the other from an unknown type of liver disease) and one (a sibling with primary biliary cirrhosis) was unavailable for testing. Our results are consistent with a familial predisposition to primary biliary cirrhosis.(ABSTRACT TRUNCATED AT 250 WORDS)


Does elective sclerotherapy improve the efficacy of long-term propranolol for prevention of recurrent bleeding in patients with severe cirrhosis? A prospective multicenter, randomized trial. We conducted a prospective, multicenter, randomized trial to compare the efficacy of sclerotherapy plus propranolol with that of propranolol alone in the prevention of recurrent gastroesophageal bleeding in severely cirrhotic patients. For 2 yr (1987 to 1988) 131 patients (96% of whom were alcoholic) with Child-Pugh class B or C cirrhosis (56% were class B and 44% were class C) were randomly assigned to one of our two treatment groups after cessation of variceal bleeding, without hemostatic sclerosis, and were observed for at least 2 yr. Treatment observance was good in 89% of cases; alcohol withdrawal was observed in 62% of cases. Sclerotherapy was performed weekly with 1% polidocanol, and variceal obliteration was obtained in 83% of cases, in a mean number of four sessions. The cumulative percentages (expressed as mean +/- S.D.) of recurrent bleeding at 2 yr were 42% +/- 6% for propranolol plus sclerotherapy and 59% +/- 6% for propranolol alone (a nonsignificant difference). Twenty-eight patients from the propranolol group but only 12 patients from the propranolol-plus-sclerotherapy group had recurrent bleeding from esophageal variceal rupture (p less than 0.01). The total number of blood units per patient with recurrent bleeding was slightly but not significantly more important in the propranolol group (8 +/- 7) than in the propranolol-plus-sclerotherapy group (5 +/- 5; p = 0.09).(ABSTRACT TRUNCATED AT 250 WORDS)


Ursodeoxycholic acid therapy in cystic fibrosis-associated liver disease: a dose-response study. Previous studies from our groups have demonstrated improvements in biochemical markers of liver function when cystic fibrosis patients with associated liver disease were administered oral ursodeoxycholic acid. The magnitude of the response was somewhat less than that found when comparable doses (10 to 15 mg/kg body wt/day) of ursodeoxycholic acid are given to other liver disease patients; this may be explained by the bile acid malabsorption that is characteristic of the disease. For this reason a dose-response study was carried out in nine cystic fibrosis patients with liver disease to establish whether improved efficacy could be obtained with higher doses. Ursodeoxycholic acid in doses of 5, 10 and 15 mg/kg body wt/day was given orally for consecutive 2-mo periods in a replicated Latin-square design. After this, all patients received 20 mg/kg body wt/day. Liver function, individual serum bile acids and biliary bile acid composition were determined at entry and at the end of each treatment period. Our data demonstrate that the magnitude of the biochemical improvement in serum liver enzymes was significantly greater with higher doses of ursodeoxycholic acid; at 20 mg/kg body wt/day it was similar to that reported for patients with other liver diseases administered lower doses. Biliary ursodeoxycholic acid enrichment increased with increasing doses, attaining 42% +/- 6% of the total biliary bile acids with the highest dose. Fasting serum ursodeoxycholic acid concentrations increased during ursodeoxycholic acid administration but were variable and correlated poorly with the dose of ursodeoxycholic acid administered, whereas no correlation was found between serum ursodeoxycholic acid concentration and the proportion of ursodeoxycholic acid in bile.(ABSTRACT TRUNCATED AT 250 WORDS)


Deficient interleukin-2 responsiveness of T lymphocytes from patients with primary biliary cirrhosis. There is increasing evidence that primary biliary cirrhosis is associated with an alteration of the immune system. Although the cause remains unknown, it has been suggested that the immune system of patients with primary biliary cirrhosis is involved in the pathogenesis of their disease. We have investigated the T-cell function in patients with primary biliary cirrhosis and have found defective phytohemagglutinin-induced T-cell mitogenesis. Likewise, their blastogenic response to CD3 monoclonal antibody was also depressed, although the DNA synthesis induced by stimulation with phorbol esters (12-O-tetradecanoil-phorbol-13-acetate) plus ionophore (ionomycin) was normal. These alterations could not be ascribed either to a decreased synthesis of interleukin-2 or to a defective expression of interleukin-2 receptor after cellular activation. Moreover, this defective proliferative response of T lymphocytes was observed even in the presence of saturating concentrations of exogenous interleukin-2. These results represent evidence of the deficiency in the interleukin-2-dependent pathway found in T lymphocytes from patients with primary biliary cirrhosis.


Effect of loxiglumide and atropine on erythromycin-induced reduction in gallbladder volume in human subjects. This study was undertaken to investigate the effect of erythromycin, a motilin agonist with prokinetic activity, on fasting gallbladder volume. To evaluate the mechanism of action of erythromycin on gallbladder motility, erythromycin (3.5 mg/kg.20 min, intravenously) was infused on three separate occasions: during cholinergic blockage with atropine (0.005 mg/kg.hr), during cholecystokinin receptor blockade with loxiglumide (10 mg/kg.hr) and during saline solution infusion (control). Atropine, loxiglumide and saline solution infusions were started 3 hr before administration of erythromycin and were continued for 3 hr thereafter. Gallbladder volumes (measured by ultrasonography), plasma cholecystokinin levels (radioimmunoassay) and plasma pancreatic polypeptide levels (radioimmunoassay) were determined at regular intervals for 6 hr in six healthy volunteers. During the 3-hr infusion before administration of erythromycin, both loxiglumide and atropine significantly increased gallbladder volumes--from 18 +/- 2 to 37 +/- 3 cm3 (p less than 0.05) and from 17 +/- 3 to 24 +/- 2 cm3 (p less than 0.05), respectively--whereas saline solution did not significantly affect gallbladder volume. During control saline solution infusion, erythromycin induced prolonged gallbladder contraction that was significant (p less than 0.05) between 60 and 180 min and reached a maximum of 45% +/- 8% at 150 min. Plasma cholecystokinin levels were not affected by erythromycin. Erythromycin induced a significant (p less than 0.05) increase in plasma pancreatic polypeptide levels, from 12 +/- 1 pmol/L to 34 +/- 3 pmol/L. Loxiglumide did not prevent the erythromycin-induced reduction in gallbladder volume. Atropine markedly reduced the effect of erythromycin, causing slight but significant (p less than 0.05) gallbladder volume reductions (18% +/- 4%) between 150 and 180 min.(ABSTRACT TRUNCATED AT 250 WORDS)


31P magnetic resonance spectroscopy detects a functional abnormality in liver metabolism after acetaminophen poisoning. Eighteen patients with acetaminophen poisoning were studied with 31P magnetic resonance spectroscopy to measure phosphorus-containing metabolites in their livers. The concentrations of all magnetic resonance-detectable metabolites fell in parallel with a decrease in the synthetic ability of the liver, indicated by the prothrombin time ratio (international normalized ratio). In particular, ATP fell to about 20% of its normal concentration in severely affected patients, as did the phosphodiester signal, which is thought to arise mainly from the endoplasmic reticulum in the liver. The correlation between ATP levels and international normalized ratio suggests that the international normalized ratio is a more accurate measure of the number of viable hepatocytes than are other biochemical tests.


Saturability of hepatic iron deposits in genetic hemochromatosis. The relationship of pretreatment serum ferritin and hepatic iron concentration to body iron removed by venesections was evaluated in 33 patients with genetic hemochromatosis. The median values of the three variables considered were 1,950 micrograms/L (range = 255 to 10,000), 1,175 micrograms/100 mg dry weight (range = 270 to 4,310) and 10 gm (range = 2 to 41), respectively. At basal liver biopsy 18 patients had cirrhosis, 6 patients had fibrosis and 9 patients had a normal pattern; siderosis was degree 3 in 6 patients and degree 4 in 27 patients. The results of fitting a polynomial regression of second degree showed that the curve of serum ferritin on iron removed was a straight line (R2 = 0.79, with a significant coefficient of linearity, p less than 0.01, and a nonsignificant coefficient of curvature), whereas that of hepatic iron concentration on iron removed showed a curvature (R2 = 0.62, with significant coefficient of linearity and curvature, p less than 0.01) and reached a plateau. The sigmoid model fit the curve of hepatic iron concentration on iron removed (R2 = 0.61), which suggested a saturation of hepatic iron storage capability; the asymptote corresponded to a hepatic iron concentration of about 2,000 micrograms/100 mg. In alcoholic patients (17 cases) the location of the sigmoid was greater than in nonalcoholic patients. Our results suggest that iron deposition occurs in the liver before other organs are involved and that with massive iron overload hepatic deposits reach saturation, after which hepatic iron concentration does not always reflect the amount of total stores. Alcohol consumption could slow the saturation of hepatic iron deposits.


Dietary N-3 polyunsaturated fatty acids decrease biliary cholesterol saturation in gallstone disease. Because fatty acid composition of biliary phospholipids influences cholesterol secretion into bile, we investigated whether replacement of n-1 monounsaturated or n-6 polyunsaturated fatty acids with n-3 polyunsaturated fatty acids in biliary phosphatidylcholines reduces supersaturation with cholesterol and prevents precipitation of cholesterol crystals in bile of gallstone patients. Seven patients with radiolucent gallstones in functioning gallbladders were studied before (control) and after 5 wk of dietary supplementation with marine fish oil (11.3 gm/day = 3.75 gm n-3 polyunsaturated fatty acids/day). Duodenal bile was collected for analysis during intravenous infusion of cholecystokinin. Gallbladder emptying in response to cholecystokinin was comparable before and during intake of n-3 polyunsaturated fatty acids. Intake of n-3 polyunsaturated fatty acids increased (p less than 0.001) the fractions of eicosapentaenoic and docosahexaenoic acids and decreased the fractions of linoleic (p less than 0.001) and arachidonic acids (p less than 0.02) in biliary phospholipids. Concomitantly, the molar ratio of cholesterol to phospholipids decreased (-19%; p less than 0.05). As a consequence, the cholesterol saturation index was reduced by -25% (p = 0.01), from 1.60 +/- 0.44 to 1.24 +/- 0.38. However, in vitro nucleation time of duodenal bile was not prolonged. The decrease in cholesterol saturation was not sufficient to prevent nucleation of cholesterol crystals in bile of gallstone patients. In conclusion, our data suggest that cholesterol saturation can be influenced by the fatty acid composition of the phosphatidylcholines secreted in bile.


Analysis of hepatocellular proliferation: study of archival liver tissue is facilitated by an endogenous marker of DNA replication. Assessment of liver regeneration with endogenous genes that are expressed during DNA replication is physiological, specific and direct. To determine whether H3 histone messenger RNA expression (which is tightly coupled with DNA synthesis) could be used for this purpose, we initially examined liver regeneration in a mouse model. After partial hepatectomy, RNA transblot studies showed induction of H3 histone messenger RNA expression in regenerating mouse livers. In situ molecular hybridization demonstrated that the overall pattern of H3 histone messenger RNA expression correlated with [3H]thymidine labeling of hepatocytes. After partial hepatectomy, H3 histone messenger RNA expression in hepatocytes peaked at 48 hr (greater than 60 times greater than at 24 hr; p less than 0.001) and then rapidly declined. Although hepatocyte labeling with [3H]thymidine showed similar kinetics of liver regeneration, use of this parameter resulted in overestimation of the proliferative compartment when it was compared with H3 histone messenger RNA expression. Next we determined whether H3 histone messenger RNA expression could be used to study hepatocellular proliferation in archival human material. H3 histone messenger RNA-expressing hepatocytes were identified on in situ hybridization in patients with acute or chronic active hepatitis and active cirrhosis, but not inactive cirrhosis. These studies demonstrate that H3 histone messenger RNA is expressed in a phasic manner during liver regeneration. Use of H3 histone messenger RNA expression to evaluate hepatocellular proliferation should facilitate clinical studies and greatly advance our understanding of the pathophysiology of liver regeneration.


Metabolic clearance rate of arginine vasopressin in patients with cirrhosis. Metabolic clearance rate and half-time of arginine vasopressin were measured in 43 cirrhotic patients and 10 control subjects. Synthetic arginine vasopressin was infused intravenously at a rate of 500 pg/min/kg of body weight for 75 min. The metabolic clearance rate was significantly reduced, and the half-time of arginine vasopressin after stopping the infusion was significantly increased in patients with cirrhosis, particularly in those with ascites and in those with moderate or severe liver dysfunction. Changes in metabolic clearance rate and half-time of arginine vasopressin correlated with the score of the liver dysfunction, prothrombin activity and levels of serum albumin and bilirubin but not with parameters of kidney function (serum creatinine levels and clearance of creatinine). We conclude that reduced metabolic clearance rate and prolonged half-time of vasopressin in plasma are frequent findings in cirrhotic patients with poor liver function. This impaired catabolism of antidiuretic hormone may contribute to maintaining elevated plasma levels of this hormone in these patients and may be an additional factor leading to fluid retention and to dilutional hyponatremia.


Acute effects of topical methyl tert-butyl ether or ethyl propionate on gallbladder histology in animals: a comparison of two solvents for contact dissolution of cholesterol gallstones. Experiments were performed in anesthetized rabbits and piglets to assess gallbladder mucosal injury during irrigation with methyl tert-butyl ether, a C5 ether, or ethyl propionate, a C5 ester--two organic solvents used in the contact dissolution of cholesterol gallstones. In 44 New Zealand White rabbits, the gallbladder was exposed to individual solvents or saline solution through a transhepatic catheter for 2 hr. Gallbladders were then harvested and fixed immediately or after a recovery period of 1, 4 or 8 days. Tissue sections were examined under light microscopy, and severity of injury was graded with predefined criteria by two pathologists blinded to the animals' treatment regimens. Histological assessment showed severe mucosal injury such as necrosis of the cells at the villus tips immediately after 2 hr of exposure to either solvent. After 4 days, injury had decreased significantly; after 8 days, complete mucosal healing had taken place. A similar study was performed in 32 piglets. Solvent or saline solution was oscillated in and out of the gallbladders of these piglets with a computer-controlled syringe pump at a pressure less than the leakage pressure of the gallbladder. Histological assessment was performed on tissue samples obtained immediately after the procedure or 8 days later. Both solvents caused severe mucosal injury; however, after 8 days complete mucosal healing had occurred, so that gallbladders exposed to solvent were indistinguishable from gallbladders exposed to saline solution, which was used as control. We conclude that both methyl tert-butyl ether and ethyl propionate cause moderate to severe epithelial injury but that the gallbladder epithelium regenerates within a few days.(ABSTRACT TRUNCATED AT 250 WORDS)


Role of cytochrome P-450 2E1 in ethanol-, carbon tetrachloride- and iron-dependent microsomal lipid peroxidation. This study investigated the role of cytochrome P-450 2E1 in enhanced microsomal lipid peroxidation in experimental alcoholic liver disease. We also examined the contribution of this isoform to the increased microsomal injury in alcoholic liver disease caused by carbon tetrachloride-induced or iron-induced oxidant stress. Adult male Wistar rats were intragastrically infused with a high-fat diet and ethanol or glucose for 16 wk; this resulted in hepatic lipid peroxidation and fibrogenesis in the ethanol-fed animals. Microsomes were isolated by differential centrifugation in the presence of 100 mumol/L deferoxamine, washed twice in buffer without deferoxamine and incubated in the absence or presence of ethanol (50 mmol/L), carbon tetrachloride (150 mumol/L), ferric citrate (50 mumol/L) or ferric citrate plus ethanol at 37 degrees C for 30 min in an NADPH-generating system. The basal rate of lipid peroxidation in microsomes isolated from ethanol-fed rats was increased by 52% compared with that in microsomes from controls. Carbon tetrachloride-induced and ferric citrate-induced lipid peroxidation were also accentuated in microsomes from ethanol-fed rats, by 76% and 108%, respectively. Ethanol added in vitro significantly reduced basal (-58%) and ferric citrate-induced (-48%) lipid peroxidation in microsomes from ethanol-fed rats, whereas it had an insignificant effect on that in control microsomes. In fact, this protective effect of ethanol on microsomes from ethanol-fed rats resulted in attenuation of the difference in the level of microsomal lipid peroxidation between the two groups.(ABSTRACT TRUNCATED AT 250 WORDS)


Hepatic accumulation of lysosomes and defective transcytotic vesicular pathways in cirrhotic rat liver. To investigate the potential role of lysosomes in cirrhosis, we analyzed the activity of lysosomal enzymes in rats exposed long-term to phenobarbital and carbon tetrachloride. The activity of lysosomal enzymes was markedly increased in the homogenate of cirrhotic livers (e.g., arylsulfatase 9 +/- S.D.2 vs. 16 +/- 6 nmoles.min-1.mg-1 in control rats and cirrhotic rats, respectively; p less than 0.001). The corresponding plasma levels were also increased (7 +/- 1 vs. 12 +/- 3 nmoles.min-1.mg-1; p less than 0.01), whereas biliary excretion was diminished (16 +/- 7 vs. 7 +/- 2 pmol.min-1.gm liver-1; p less than 0.05) in cirrhotic rats. Stereological quantification of lysosomes visualized cytochemically revealed an increase of pericanalicular lysosomes averaging 1.5 +/- 0.4 around a canaliculus in controls and 3.7 +/- 1.0 in cirrhotic rats (p less than 0.01). Because this suggested a defect in the transcellular vesicular pathway, we investigated the biliary excretion of horseradish peroxidase and epidermal growth factor in perfused livers. Bile flow and total horseradish peroxidase excretion were similar in control rats and cirrhotic rats. However, the early peak of biliary horseradish peroxidase excretion--usually taken as evidence of paracellular transport--was increased in cirrhotic rats (13 +/- 7 vs. 57 +/- 22%; p less than 0.01), whereas the second peak--reflecting the transcellular vesicular pathway(s)--was markedly reduced (87 +/- 7 vs. 43 +/- 22%; p less than 0.001). A similar reduction in the biliary excretion of intact epidermal growth factor and of its degradation products was found. These results demonstrate an increased number of lysosomes in hepatocytes of cirrhotic livers; this appears to be the result of accumulation rather than proliferation, in view of the reduced transcellular vesicular movement of different markers into bile.


Detection of immunoglobulin gene rearrangement in B-cell lymphomas by polymerase chain reaction gene amplification. This is a report on our attempt to use polymerase chain reaction (PCR) to detect rearrangement of the immunoglobulin gene in the tissue specimens obtained from 30 patients with non-Hodgkin's lymphomas. There were 20 B-cell lymphomas and 10 T-cell. All 20 B-cell lymphomas but none of the 10 T-cell lymphomas had JH rearrangement by Southern analysis. Two pairs of primers (V670/OL-4 and VH26/OL-4) were designed to amplify the CDR3 region of the immunoglobulin gene heavy chain. The PCR analysis was positive using either one or both pairs of primers in 11 of the the 20 cases (55 per cent) of B-cell lymphomas which all had positive rearrangement by Southern analysis. The two pairs of primers seemed to produce complementary results as the specimens may be positive to one pair but negative to the other. The false negative rate of 45 per cent is however much higher than the respective figures of 18 per cent and 0 per cent observed in our patients with acute lymphoblastic leukemia and chronic lymphocytic leukemia in a previous study. Peripheral blood and bone marrow biopsy specimens obtained at the time of initial diagnosis were available from 10 patients with B-cell lymphomas whose lymph node biopsy specimens at the time of diagnosis were positive by both Southern analysis and PCR. All these peripheral blood and marrow specimens had no microscopic evidence of involvement by lymphoma cells and JH rearrangement was not detected by Southern analysis. However, rearranged bands identical to that of the lymph node biopsy specimen were detected by PCR in the peripheral and marrow blood of one of them. This PCR technique has been shown to have a sensitivity of 0.1 per cent in our previous report and may be more useful than morphology alone or Southern analysis in detecting minimal lymphomatous involvement in the peripheral blood and bone marrow at the time of initial diagnosis. Further clinical correlation is required to confirm the finding.


Diagnostic difficulties in 'non-mycotic' cutaneous lymphoproliferative diseases. The diagnosis of cutaneous lymphoproliferative diseases is an area of bewildering complexity to many histopathologists. This article concentrates on 'non-mycotic' cutaneous diseases. The 'current state of the diagnostic art' is critically assessed. Cutaneous 'pseudolymphoma' is relegated to the position of an aid-memoire and is not a diagnosis. Inadequacies in the classification of cutaneous lymphoma are discussed and the non-specificity of many histopathological features is highlighted. The status of specific entities is analysed and the contribution of modern investigative techniques in diagnosis is evaluated. This includes cutaneous T-cell lymphomas with a detailed consideration of large cell lymphoma heterogeneity. Cutaneous B-cell diseases are shown to be an unresolved diagnostic maze and the necessity for new clearly defined diagnostic criteria is emphasized. Evidence is presented to show that many cutaneous lymphoproliferative diseases lie on continuous spectra that, initially, are probably antigenically driven, and that a diagnosis is best achieved by a multifaceted approach. This is exemplified by cutaneous diseases that have origins from both B- (cutaneous lymphoid hyperplasia/lymphoma) and T-cells (lymphomatoid papulosis, lymphomatoid granulomatosis and mycosis fungoides). The future diagnostic role of the polymerase chain reaction and cytogenetic analysis is discussed. Intriguingly, recent molecular evidence has shown that lymphomatoid papulosis, cutaneous T-cell lymphoma, CD30 positive large cell lymphoma and Hodgkin's disease can originate from a single T-cell clone and display an identical chromosomal translocation and T-cell receptor rearrangement. Careful clinico-pathological correlation combined with prolonged patient follow-up remains the gold standard for diagnosis.


The prognostic factors for peripheral T-cell lymphomas. The peripheral T-cell lymphomas of 50 patients were classified by histological type and tumour cell size, the latter as described by the Nebraska Group. There was no difference in the pattern of their clinical features among the various histological subtypes and their clinical outcome was similar. Cell size and other factors including sex, age, presence of B symptoms, sites of primary disease, histological subtypes according to the Japanese classification, lactate dehydrogenase level and presence of bulky disease did not appear to correlate with clinical outcome. However, patients with stage IV disease had significantly lower CR rate (9/29 versus 16/21, p = 0.01) and poorer survival at 3 years (p = 0.05) than those with stage I, II and III disease. Patients receiving COPP, the less intensive chemotherapy regimen, appeared to have poorer clinical outcome but the difference observed was not statistically significant because of the number of patients in each subgroup.


Stereological estimates of nuclear volume in carcinoma of the ampulla of Vater. The stereological estimate of mean nuclear volume is an objective and reproducible method of measurement of nuclear size in terms of absolute volume. We have used this method to study carcinomas of the ampulla of Vater. Our study includes 21 cases--five papillary and 16 intestinal--all treated by the same surgical procedure. The volume-weighted mean nuclear volume (nuclear nu v) and the mean nuclear area were calculated. The mean volumes of nuclear nu v were significantly different (P less than 0.01) between the papillary and intestinal carcinomas, but no differences were found between normal mucosa and papillary carcinoma. The variance was significantly larger in the intestinal group than the papillary and normal groups. The nuclear nu v showed a significant association with survival, larger nuclear nu v (greater than 150 microns 3) being associated with a lower survival rate.


A major solid undifferentiated carcinoma pattern correlates with tumour progression in locally advanced prostatic carcinoma. Solid undifferentiated carcinoma was the major microscopic pattern in 24 prostatic carcinomas, 12 of which were clinically recurrent. Tumour cells were uniform, with moderately hyperchromatic nuclei and indistinct cytoplasm, and were arranged in solid or focally irregular aggregates. In areas, the tumour cells were large with vesicular nuclei, nucleoli and more abundant cytoplasm. In previous specimens, solid undifferentiated carcinoma was absent or was a minor pattern. Twenty of 23 cases showed prostate specific antigen and prostatic acid phosphatase immunoreactivity, and nine of 17 cases contained scattered argyrophilic or chromogranin-immunoreactive cells. On proliferating cell nuclear antigen immunostaining of 12 specimens, the mean tumour proliferative fraction in solid undifferentiated carcinoma (range: 10.5-18%) was greater than in areas of grade 3 prostatic carcinoma (range: 3-6%). In all 22 stage C and D cases, there was a close correlation with clinical evidence of tumour progression, and the overall 2-year survival rate was only 16.7%. It is concluded that a major solid undifferentiated pattern correlates with increased biological aggressiveness and a poor prognosis in locally advanced prostatic carcinoma.


Angiocentric T-cell lymphoma of the nose, paranasal sinuses and hard palate. Sixty-five cases of malignant lymphoma of the nose, paranasal sinuses and hard palate were retrospectively analysed to identify the presence or absence of angiocentric lesions. We observed that the 23 patients with angiocentric lesions had a worse prognosis with a shorter duration of response and also a shorter duration of survival, compared with 42 cases of malignant lymphoma of the same anatomical region but without angiocentric lesions. Patients with angiocentric lymphoma were associated with other bad prognostic factors such as elevated levels of lactic dehydrogenase and beta 2 microglobulin, local bone destruction and lymphopenia. Immunophenotyping studies showed that most patients with angiocentric lesions had T cell lymphomas (18 of 23, 78 per cent). We believe that patients with angiocentric T cell lymphomas of the nose, paranasal sinuses and hard palate represent a distinctive clinico-pathological entity with different clinical presentation and outcome. Patients with angiocentric T cell lymphomas had frequent relapse at extranodal sites and combined therapy should be considered as the initial therapeutic approach.


Intra-patient variation between breast cancer axillary lymph node metastases using quantifiable features. The intra-patient variations of some clinically relevant quantifiable features, between axillary lymph node metastases were evaluated in 44 breast cancer patients. In all lymph node metastases detected (range 2-33 per patient), the mitotic figures were counted, the volume percentage epithelium was assessed and the mean nuclear area was measured. The intra-patient variation for each quantifiable feature was expressed by the coefficient of variation (CV). Since the measurement techniques used introduce a certain, well known variation themselves because of sampling and measurement errors, the CVs found had to be greater than methodological tolerance limits (established in previous studies) to be interpreted as indicating biological variation. The CVs exceeded the methodological tolerance limits in 86% of the cases for the mitotic count, in 48% of the cases for the volume percentage epithelium, and in 47% of the cases for the mean nuclear area. This indicated that in these cases, the variation found in the quantifiable features could not be explained by sampling or measurement errors and should be regarded as real biological variation. Furthermore, the variation in the quantifiable features studied showed a significant positive correlation with the number of lymph node metastases. Thus, there may be considerable intra-patient variation in quantifiable features between axillary lymph node metastases in breast cancer. This may indicate that these lymph node metastases originate independently from different clones within the primary tumour, that they are independently formed in different stages of tumour development, or that they, as an expression of intrinsic tumour heterogeneity, may develop in different directions from the start.


Solitary plasmacytoma of bone: clinical features, treatment and survival. Eleven patients with solitary plasmacytoma of bone were seen between 1978 and 1991. A retrospective review of the clinical features, treatment and survival of these patients was made with the aim of helping to define those at risk for early development of myeloma. Nine patients (82 per cent) had paraparoteinemia at diagnosis. Treatment consisted of local irradiation plus or minus surgery (nine patients) and surgery alone (two patients). No patient received adjuvant chemotherapy. One patient had evidence of both generalized osteopenia and immunoparesis, and progressed to myeloma within six months. Four patients (36 per cent) progressed to myeloma. All of these had paraprotein levels which continued to rise following initial treatment. Three patients died of myeloma at 12, 81 and 144 months and the other patient is alive with myeloma at 76 months. Patients without paraproteinemia at presentation or whose paraprotein decreased after treatment did not progress to myeloma. Three patients have been followed for 8, 11 and 19 years with no evidence of myeloma. Failure of paraprotein to clear after local treatment suggests occult disseminated disease and is predictive of later development of overt myeloma.


Proliferation in non-Hodgkin's lymphomas as determined by immunohistochemical double staining for Ki-67. The authors have studied proliferation (growth fractions) in non-Hodgkin's lymphomas (NHL) using an immunohistochemical double staining technique with the monoclonal antibody (MoAb), Ki-67, in order to clarify the relationships between histologic type, proliferation and prognosis. The percentages of Ki-67 positive (Ki-67+) cells in B cell lymphomas were higher for diffuse lymphomas than for follicular lymphomas and increased in order from small to large cell types. In addition, the percentage of Ki-67+ cells in B cell lymphomas inversely correlated with survival in months (n = 33 r = -0.54 p < 0.01). These results indicate that the percentage of Ki-67+ cells in B cell lymphomas correlates with histologic type and prognosis. Although the prognosis of T cell lymphomas is considered worse than that of B cell lymphomas, the percentage of Ki-67+ cells was lower, in general, in T cell lymphomas than in B cell lymphomas. These data indicate that proliferation in T cell lymphomas does not correlate with survival.


In vitro differentiation of a Hodgkin's disease derived cell line. We have examined the Hodgkin's disease derived cell line Co in terms of its capacity to differentiate in vitro. Co cells show the characteristics of immature T cells and express CD3 molecules in the cytoplasm. On activation with 12-O-tetradecanoylphorbol-13-acetate (TPA) these cells express the CD3 antigen and the T cell receptor alpha beta (TCR alpha beta) on the cell surface. Surface expression of the activation marker CD25 (IL2 receptor) was also greatly increased, whereas CD4 and CD8 levels were not altered. Supernatants of TPA-stimulated Co cells contained the cytokines IL2, IL3, IL4 and IL8, whereas these cytokines were not detected in the supernatants of untreated cells. Different subclones of the Co cell line differed in their response to TPA with respect to the induced CD3 and TCR expression. Our data demonstrate that a Hodgkin's disease derived cell line can be induced to differentiate in vitro from a pre-T cell phenotype towards a more mature T cell. It is possible that similar processes may occur in Hodgkin's disease in vivo.


'Aggressive' low grade lymphocytic lymphomas can be identified by flow cytometric S-phase determinations. Forty-five patients with low grade non-Hodgkin's lymphomas were studied with respect to the fraction of S-phase cells in fresh tumour material by flow cytometric analysis. Patients with stage I lymphomas were treated with radiotherapy, patients with stage II-IV lymphomas with Prednimustine (Sterecyt). Patients with lymphocytic lymphomas of CLL type were only treated if they had symptoms. Median S-phase fraction in the samples was 2.0 per cent. A significantly shorter survival was found for patients with lymphocytic lymphomas with S-phase fractions > 2.0 per cent compared with cases showing lower S-phase fractions. No significant difference in survival was found in the subgroups of immunocytic or follicular and follicular/diffuse centroblastic/centrocytic lymphomas. In a Cox multivariate analysis, in which also age, constitutional symptoms, stage and morphology were included, the fraction of S-phase cells was found to be a statistically significant, prognostic parameter for low grade lymphomas, mainly due to the result in the subgroup of lymphocytic lymphomas.


Thymic carcinosarcoma associated with a spindle cell thymoma: an immunohistochemical study. A case of thymic carcinosarcoma associated with a spindle cell thymoma in a 71-year-old woman is reported. Histological and immunohistochemical studies of the carcinosarcoma showed two quite different components: the sarcomatous component included cells with myoid differentiation which stained for desmin and muscle specific actin, and some isolated cells which stained positively for low molecular weight cytokeratin, while the carcinomatous component, which formed less than 10% of the tumour, showed an epithelial phenotype, being positive for low and high molecular weight cytokeratin and epithelial membrane antigen. The thymoma cells showed epithelial markers, and a few cells were also positive for desmin and muscle specific actin. The rarity of this tumour and its possible histogenesis are discussed.


Acute myelogenous leukemia (AML) and diabetes insipidus (DI): further association with monosomy 7. Diabetes insipidus (DI) is a rare complication of acute myelogenous leukemia (AML). Five of the six such cases described in the literature who had banded chromosomal studies performed had monosomy 7 and the sixth patient had del(7)(q22). A further case of AML complicated by DI in whom banded chromosomal studies revealed a complex karyotypic abnormality, including monosomy 7, is reported. The association between monosomy 7 and DI in AML appears specific but the reason for this association remains unclear.


Design of an expert system and its application to dermatopathology. Expert systems are computer programs which use inference and knowledge to solve problems which usually require the expertise of a human specialist. This paper examines the application of expert systems to histopathology and explains their construction by describing the design of an expert system 'dermdx', intended to aid in the interpretation and diagnosis of biopsies of inflammatory diseases of the skin. The system consists of an expert shell, which performs the inference, and a rule-base, which contains the knowledge with which the system operates. The system can be easily updated or adapted to other tasks.


Bone mineral density (BMD) in patients with lymphoma: the effects of chemotherapy, intermittent corticosteroids and premature menopause. Young women with a chemotherapy-induced early menopause are theoretically at considerable risk of developing post-menopausal osteoporosis with problems developing earlier and more severely. In this study bone mineral density (BMD) measurements were made, using a dual-energy X-ray absorptiometer (DXA), at the spine and hip of 50 young women who had been treated for lymphoma, 24 of whom were post-menopausal and 78, healthy age-matched controls. On analysis of the results, there was no significant difference between the control group and the 26 post-treatment, pre-menopausal patients, but the BMD levels were significantly lower than the controls in the post-menopausal group particularly in 16 patients who had been menopausal greater than 18 months. The results confirm that these young women with treatment-induced premature menopause are at considerable risk of developing osteoporotic problems. Early recognition of this is important so that preventative measures with hormone replacement therapy can be initiated where this is safely possible. The results also indicate that chemotherapy for lymphoma (cytotoxics and high dose intermittent steroids), are unlikely to contribute directly to the lowering of the BMD of these patients.


Sebaceous glands and hair follicles in the cervix uteri. We report a case of ectopic ectodermal structures in the ectocervix of a 51-year-old female. Sebaceous glands and numerous abnormal hair follicles were present in the stroma of an otherwise normal cervix.


Verapamil preferentially potentiates in-vitro cytotoxicity of vincristine on malignant lymphoid cells. Verapamil has been shown to overcome acquired drug resistance to vincristine in P388 leukemia both in vitro and in vivo. To study the selectivity of this action, the effect of addition of verapamil on the cytotoxicity of vincristine was studied using lymphocytes from eight patients with chronic lymphocytic leukemia (CLL), lymphoblasts from a T-acute lymphoblastic leukemia (T-ALL) cell line (GM 3639), and peripheral blood lymphocytes (PBL) from eight normal healthy volunteers. Using the differential staining cytotoxicity (DiSC) assay, we demonstrated that verapamil at 1 microM concentration potentiated the in-vitro cytotoxicity of vincristine on CLL and GM 3639 cells in concentrations of 0.04-0.25 micrograms/l. There was however, no enhancement of cytotoxicity noted against the control PBL. The data demonstrate that verapamil preferentially enhances the in-vitro cytotoxicity of vincristine on CLL and GM 3639 cells but no enhancement of cytotoxicity is seen against PBL.


Necrotizing sialometaplasia (adenometaplasia) of the trachea. Necrotizing sialometaplasia is a benign condition first described in minor salivary glands of the soft palate with morphological changes which can be misinterpreted as squamous-cell carcinoma. Similar lesions have been subsequently reported in other locations including major salivary glands, lip, breast and skin (the term syringometaplasia has been applied for the latter). We report three cases of such a process involving submucosal glands in the trachea following prolonged translaryngeal intubation.


Ceroid granuloma of the uterine cervix.

Ceroid granuloma of the uterine cervix. An excision biopsy was taken from a 59-year-old woman with a small dark-brown lesion on the anterior lip of the cervix. The histology revealed an ulcerated surface epithelium with a band-like infiltrate of pigment containing macrophages in the subepithelial zone. Histochemical examination of the specimen revealed that the pigment was ceroid. This is, to the best of our knowledge, the first case report of ceroid granuloma of the uterine cervix.


Recombinant DNA technology and its diagnostic applications. As yet recombinant DNA technology does not appear to have widespread diagnostic application in pathology. However, it does have a useful role to play in specific circumstances in at least three main areas: a it can provide precise diagnostic information about genetic diseases, allowing appropriate counselling, and indicating future directions for research on therapeutic intervention, e.g. gene therapy; b micro-organisms can be identified more sensitively and specifically, in fresh or fixed tissue samples, and their genomes can be analysed in fine detail, providing information relevant to the aetiology, epidemiology and pathogenesis of many diseases; c in tumour pathology the main application so far has been to resolve diagnostic problems associated with leukaemias and lymphomas, when other diagnostic procedures have been inconclusive. Specific chromosomal translocations, involving recognized genes, are particularly amenable to diagnosis by these means. Diagnostic applications to solid tumours are yet to be identified, although significant insights into tumorigenesis have been obtained, and these may ultimately lead to the development of useful markers for prognostic and therapeutic purposes.


Pathogenesis of congenital cystic adenomatoid malformation of the lung. Congenital cystic adenomatoid malformation is a rare developmental abnormality of the lung. In most earlier reported cases, the anatomy of the bronchial tree was poorly documented. We describe four cases studied following autopsy. Post-mortem bronchography or serial microscopical examination showed segmental bronchial absence or atresia in each of them. Our observations provide further evidence pointing to bronchial atresia as being the primary defect leading to the development of congenital cystic adenomatoid malformation. The morphology of the lesion, i.e. the type of malformation, is determined by the extent of dysplastic lung growth beyond the atretic segment. The aetiology of the bronchial atresia is probably heterogeneous and may either represent a primary malformation, or be the result of vascular disruption.


p53 expression is common in malignant mesothelioma. The p53 tumour suppressor gene has been shown to be frequently mutated in a wide range of human neoplasms. This is accompanied by increased levels of p53 protein which become immunologically detectable in pathological material. We have investigated the possibility that the differential diagnosis between reactive and neoplastic mesothelium might be resolved using a polyclonal serum raised to human p53 protein, CM-1. None of 20 cases of reactive mesothelial proliferation showed p53 immunoreactivity while 70% (14 of 20) of cases of malignant mesothelioma showed p53 staining. We can thus infer that abnormalities of p53 appear to be a common event in malignant mesothelioma and that p53 immunostaining may be of value in the distinction of malignant mesothelioma from reactive hyperplasia.


Light microscopic and ultrastructural distribution of type VI collagen in human liver: alterations in chronic biliary disease. We have investigated the distribution of type VI collagen in normal human liver obtained from cadaveric renal transplant donors, using a peroxidase-antiperoxidase method for light microscopic visualization, and an immunogold labelling method for ultrastructural localization. The distribution was compared with that of the more abundant interstitial collagen type III, using antibodies to amino terminal procollagen type III. Staining for type VI collagen was identified in Glisson's capsule, in portal tract stroma and within the space of Disse. Perisinusoidal staining showed intra-acinar heterogeneity with the intensity in acinar zones 2 and 3 being greater than in zone 1. Type III collagen was also found in the space of Disse although no significant intra-acinar variation in staining intensity was noted. Immuno-gold labelling for type VI collagen was demonstrated on amorphous or microfilamentous material lying between, and occasionally appearing to interconnect, cross-striated collagen fibrils, whereas labelling for amino terminal procollagen type III was exclusively on fibrils. Intracellular staining for type VI collagen was noted in perisinusoidal (lto) cells. These results confirm that type VI collagen is a ubiquitous constituent of the normal hepatic extracellular matrix and suggest that it may be synthesized by perisinusoidal (lto) cells. The distribution of type VI collagen was also studied in biopsy material from patients with different histological stages of primary biliary cirrhosis. Intense staining was noted around proliferating bile ductules within developing fibrous septa and in established septa of cirrhotic liver. These observations indicate that this 'minor' matrix component may play an important role in hepatic fibrogenesis.


Collagenous and basement membrane proteins of chordoma: immunohistochemical analysis. Tissue localization of collagenous and basement membrane proteins in the extracellular matrix of five sacro-coccygeal chordomas and human fetal notochords was examined immunohistochemically to assess the implications for the histogenesis and histological diagnosis of chordoma. Human fetal notochords and conventional chordomas both exhibited basement membrane proteins (such as type IV collagen and laminin) and type VI collagen on the surfaces of cellular cords. Type II collagen, a main structural protein of cartilage, was also present in both tissues. In the chordomas, however, type II collagen was not so widespread as it was in the notochords, and the predominant collagenous protein was type I. In contrast, an altered deposition of these proteins was noticed in a recurrent tumour which, histologically, showed considerable atypia and eventually metastasized to the liver. The characteristic cartilage-type and basement membrane proteins disappeared and unusual collagen types, such as types III and V, appeared in the stroma. The results further support the notochordal origin of chordoma and suggest that the immunohistochemistry of collagenous and basement membrane proteins may be a helpful criterion for the histological diagnosis and prediction of the biological aggressiveness of chordomas.


Detection of immunoglobulin light chain mRNA in nodular sclerosing Hodgkin's disease by in situ hybridization with biotinylated oligonucleotide probes compared with immunohistochemical staining with poly- and monoclonal antibodies. In order to elucidate the origin of the Hodgkin's and Reed-Sternberg cells, the expression of immunoglobulin kappa- and lambda light chain mRNA in 23 cases of nodular sclerosing and two cases of mixed cellularity Hodgkin's disease was examined by in situ hybridization using biotinylated oligonucleotide probes and compared with immunohistochemical staining with mono- and polyclonal antibodies against immunoglobulin kappa- and lambda light chains. No hybridization signals were seen in Hodgkin's or Reed-Sternberg cells in any of the cases. Polyclonal staining with polyclonal anti-immunoglobulin light chain antibodies was seen in Hodgkin's and Reed-Sternberg cells in 12 cases of nodular sclerosis and in two cases of mixed cellularity and with monoclonal antibodies in three cases of nodular sclerosis, but in no cases of mixed cellularity. In all cases, there was polyclonal labelling of plasma cells with both the oligonucleotide probes and the antibodies. In five cases, the Hodgkin's and Reed-Sternberg cells were also stained with one of the B-cell antibodies L26, MB2 or LN1. Lack of mRNA signals in Hodgkin's and Reed-Sternberg cells might indicate that these cells in Hodgkin's disease of the nodular sclerosis subtype are either not B-cell derived or they are early B-cells (precursor B-cells) not yet able to produce immunoglobulin light chain mRNA, at least not at a level detectable by in situ hybridization. Immunohistochemical staining of Hodgkin's and Reed-Sternberg cells, however, with antibodies against immunoglobulin kappa and lambda light chains may be explained by cellular uptake of the light chains, but the difference in reactivity between poly- and monoclonal antibodies cannot be explained at present.


Focal oxyntic gland atrophy with endocrine cell hyperplasia in Zollinger-Ellison syndrome during omeprazole treatment. Development of focal gland atrophy of the oxyntic mucosa was found in two patients with the Zollinger-Ellison syndrome undergoing long-term treatment with omeprazole. The atrophic areas revealed florid proliferation of endocrine cells in the form of both intraglandular crescents and micronodular hyperplasia. This proliferation was significantly more pronounced than in the remaining non-atrophic mucosa. The possible relationship of these changes to long-standing pharmacological therapy for gastric acid suppression is discussed.


Antiphonal duetting and sex hormones in the tropical bush shrike Laniarius funebris (HARTLAUB). We investigated changes in antiphonal duetting with phases of reproduction and circulating levels of luteinizing hormone, testosterone, and estradiol in slate-colored boubous (Laniarius funebris) breeding in aviaries. Frequency of overall male singing did not vary with reproductive phase while frequencies of female singing and female vocal responses to male song were reduced during incubation and feeding of nestlings. This resulted in significant changes in frequency of duetting. Males sang the sexual song type M1 more often during courtship and nest building than during the nestlings phase. Their territorial song types M2 and M4 did not vary with breeding phase. Females were less responsive to M1 during incubation and to M2 during the nest building and nestlings than during the courtship phase. Plasma levels of luteinizing hormone and testosterone increased in males from the prebreeding to the courtship phase. While testosterone decreased already during nest building and remained low during subsequent phases of reproduction, luteinizing hormone decreased during incubation and feeding of nestlings. Female luteinizing hormone levels were highest during nest building. Female estradiol levels decreased from nest building to incubation and increased again during subsequent nest building. Female testosterone levels were low but not basal and did not vary with phase. Neither the overall male and female singing frequencies nor the frequencies of male song types were correlated with hormonal state. However, female participation in territorial duets M4 correlated positively with their testosterone levels. It is suggested that in this monogamous, duetting species with prolonged pairbonds behavioral cues between the mates are more important than the hormonal state in control of male and female singing.


Social influences on reproductive development and fertility in female Djungarian hamsters (Phodopus campbelli). Social influences on the sexual maturation of female Djungarian hamsters were investigated in two experiments. In the first experiment females were housed from weaning with an adult male, by themselves, or with a weanling sister. Maturation was accelerated in females housed with males as indicated by younger age at first ovulation, increased rates of ovarian and uterine growth, and lower LH levels at some ages. Maturation was delayed in females housed with sisters compared to those housed alone as measured by time of first ovulation and by lower estradiol levels at some ages. The most marked differences between groups occurred 8 to 12 days after weaning, suggesting that events during this period are particularly important in the social mediation of sexual maturation. In the second experiment the effects of reproductive suppression (caused by living with a sister) on the subsequent fertility of females housed with males were examined. If male-female pairs were housed in clean cages, no effects were observed; however, pairs housed in cages previously soiled by the female and her sister had fewer young surviving until 1 week of age despite no differences in the age of pregnancy onset or in the initial litter size. Thus, even cues present in unrenewed soiled bedding may have subtle but long lasting effects on reproductive function.


Influences of exogenous epinephrine on two reproductive parameters in female mice: disruption of receptivity but not implantation. Diverse stressors impede female receptivity and fertility. Since epinephrine is released from the adrenal during stress, it might play a role in stress-induced disruptions of female reproductive parameters. Experiment 1 examined whether exogenous epinephrine would disrupt lordosis behavior in estrogen- and progesterone-treated ovariectomized female mice. Single dosages of 20 and 40 micrograms almost eliminated sexual receptivity. Experiments 2 and 3 examined whether chronic dosages of epinephrine shortly after insemination would lead to a failure of implantation of mouse embryos. Results indicated that epinephrine had little effect on the number of dams delivering pups, the number of pups born, litter weight, or the number of stillbirths.


Development of partner preferences in female prairie voles (Microtus ochrogaster): the role of social and sexual experience. Prairie voles (Microtus ochrogaster) exhibit a monogamous mating system characterized by long-term pair bonds between mates. The purpose of this study was to examine the effect of cohabitation time and sexual experience on the development of pair bond formation in female prairie voles. Females that were allowed to cohabit for 24 hr or more, with or without mating, exhibited a strong social preference for a familiar partner versus a strange male. Females that cohabited and mated for 6 hr showed strong preferences for a familiar partner, while cohabitation for less than 24 hr, without mating, did not result in preferences for the familiar male. These results indicate that mating was not essential for partner preference formation; however, preferences developed more rapidly when mating occurred.


Role of glucocorticoids in the stress-induced suppression of testicular steroidogenesis in adult male rats. We have examined the role of glucocorticoids in the stress-induced inhibition of testicular steroidogenesis. Immobilization (3 hr) reduced plasma testosterone (T) levels to 24% of control values but did not affect plasma LH levels. This reduction was partially reversed by in vivo injections of the antiglucocorticoid, RU486, prior to the stress session at a dose of 10 mg/kg BW, but not at 1.0 or 50 mg/kg BW. Stressed rats that were treated with 10 mg/kg BW RU486 had twofold higher plasma T levels than vehicle-treated stressed animals. Injections of RU486 did not affect plasma LH levels in control or stressed rats and did not affect T levels of unstressed rats. Stressed rats had eightfold higher plasma corticosterone levels than controls, and RU486 had no effect on control or stress levels of corticosterone. The possible role of glucocorticoids in mediating the effect of stress on testicular T production was investigated also in vitro by incubating testicular interstitial cells from unstressed rats for 3 hr with corticosterone (0, 0.01, 0.1, or 1.0 microM) or dexamethasone (0, 0.001, 0.01, or 0.1 microM), followed by an additional 2 hr with hCG (0, 25, 50, or 100 microIU). Both corticosterone and dexamethasone inhibited hCG-stimulated T production in a dose-dependent manner. Cells incubated with the highest concentration of either of the glucocorticoids showed significantly reduced responses to hCG stimulation. In the absence of hCG, in vitro T production was not affected by dexamethasone or 0.01 and 0.1 microM corticosterone. However, the highest dose of corticosterone (1.0 microM) produced a 63% elevation in basal T production. Coincubation of testicular interstitial cells with corticosterone (1.0 microM) or dexamethasone (0.1 microM) and RU486 (0.01, 0.1, and 1.0 microM) reversed the glucocorticoid-induced suppressions of T production in a dose-dependent manner. Our results suggest that during stress increases in plasma levels of glucocorticoids in male rats act via glucocorticoid receptors on testicular interstitial cells to suppress the testicular response to gonadotropins, and that the decline of testosterone production during immobilization stress is in part mediated by a direct action of glucocorticoids on the testis.


The effect of estrogen treatment on scopolamine inhibition of lordosis. Previous evidence indicates that the cholinergic muscarinic antagonist, scopolamine, inhibits lordosis in female rats. In the experiments reported here, the effects of various doses and repeated administrations of estrogen on the scopolamine inhibition of lordosis were examined. In the first experiment, intraperitoneal injections of scopolamine (1 mg/rat) completely inhibited lordosis in ovariectomized rats primed with low doses of estradiol benzoate (0.25 or 0.5 micrograms for 3 days) and progesterone (500 micrograms). However, scopolamine was significantly less effective in inhibiting lordosis in females primed with a higher dose of estradiol benzoate (25 micrograms for 3 days) and progesterone (500 micrograms). When hormone priming was repeated on subsequent weeks, scopolamine continued to inhibit lordosis in females that received 0.25 micrograms estradiol benzoate but was less effective in females primed with 0.5 micrograms. Scopolamine failed to inhibit lordosis in females treated with 25 micrograms estradiol benzoate on these later tests. In the second experiment, various doses of scopolamine (1, 2, or 4 mg/rat) were administered intraperitoneally to females primed with the highest dose of estradiol benzoate (25 micrograms) and progesterone (500 micrograms). Lordosis was inhibited equally by all scopolamine doses during the first week. As in the first experiment, scopolamine failed to inhibit lordosis at all doses on subsequent weeks of testing. These results indicate that the ability of scopolamine to inhibit lordosis is reduced by increasing the dose or the number of estrogen exposures. Because higher doses of scopolamine failed to restore its inhibitory effect on lordosis an upregulation of muscarinic receptors by estrogen cannot account for the reduced effectiveness of scopolamine.


Behavioral and cardiac responses after intracerebroventricular corticotropin-releasing hormone (CRH) administration: role of adrenal cortical hormones. Intracerebroventricularly (icv) administered corticotropin-releasing hormone (CRH) produces a dose-dependent increase in heart rate in association with behavioral activation. The present study was designed to investigate whether these CRH-induced responses are dependent on adrenal function. The effects of adrenalectomy (ADX) and subsequent corticosterone replacement were studied. Administration icv of 300 ng of CRH failed to produce behavioral activation and tachycardia in ADX rats. Corticosterone replacement restored the CRH-induced behavioral response to preoperative levels, whereas the CRH-induced tachycardia was partially restored. This latter result may be related to the fact that the baseline heart rate of ADX animals appeared to be significantly higher than that of corticosterone-treated ADX animals. It is concluded that circulating adrenal corticosterone in ADX rats is involved in the expression of the behavioral and cardiac effect of central CRH.


Fear and distress in Japanese quail chicks of two lines genetically selected for low or high adrenocortical response to immobilization stress. Behavioral and adrenocortical reactivity to stressful stimulation was examined in 12- and 13-day-old chicks of two lines of Japanese quail selected over several generations for exaggerated (HS: high stress) or reduced (LS: low stress) plasma corticosterone (B) response to brief immobilization stress. Plasma B concentrations and tonic immobility (TI) fear reactions were measured in unstressed (control) and stressed (overnight cooping) chicks of both lines. The stress treatment was applied over a period of 12-20 hr and it involved capture by the experimenter, inescapable exposure to an unfamiliar environment and to strange conspecifics, reductions in ambient temperature and floor space, and the deprivation of food and water. Chicks of the HS line were more susceptible to the induction of TI and they remained immobile longer than did LS chicks. Therefore HS chicks were considered to be more fearful than their LS counterparts. Stress treatment elicited a marked adrenocortical response that was more pronounced in HS than in LS chicks. Stress treatment also increased susceptibility to TI but did not significantly affect the duration of immobility. These findings suggest that selecting the quail for differential corticosterone response to a particular stressor had exerted an unconscious and concomitant effect on underlying fearfulness as well as on their adrenocortical reactivity to several other stressful situations. The results are further discussed in terms of a putative relationship between adrenocortical activation and fearfulness.


Testosterone and opioids interact to regulate feeding in a male migratory songbird. A male migratory songbird (dark-eyed junco, Junco hyemalis) was used as a model for studies on the influence of testosterone (T) on feeding, and on interactive effects on this behavior between T and the opioid antagonist naloxone hydrochloride (Nal). Administered chronically to birds exposed to nonstimulating photoperiods, T increased food intake by 30-58% without altering the body mass, the fat index, or the standard metabolic rate. An intramuscular injection of Nal decreased feeding temporarily in a dose-related manner. T-treated juncos exhibited a decreased sensitivity to the anorexic influence of Nal administration, demonstrating that T interacts with opioids to control food consumption. Neuroendocrine mechanisms that potentially account for this interaction are discussed.


Erectile function and bulbospongiosus EMG activity in estrogen-maintained castrated rats vary with behavioral context. Electromyographic (EMG) activity in the bulbospongiosus muscles (BS) was recorded to monitor potential castration-induced alterations in muscle activity during copulation and reflexive erections. EMG recordings were made from intact male rats and from castrated rats maintained from 7 to 50 days on estradiol benzoate (300 micrograms/day) or testosterone (200 micrograms/day). Despite a 40-50% postcastration reduction in the weight of the BS and accessory sexual glands in estrogen-treated rats, the pattern of EMG activity during copulation was similar across groups. In estradiol-treated males, the EMG burst frequency during mounts and burst duration during intromissions exceeded the parameters of intact males and of castrated males maintained on testosterone. Between intromissions, and following ejaculatory patterns, estrogen-treated males displayed spontaneous muscle bursts accompanied by visually confirmed erection of the glans penis, but these males quickly lost the capacity for reflexive erections. These data demonstrate that despite castration-induced atrophy of the penile muscles and, presumably, their spinal motor nuclei, the motor output to these muscles is maintained following androgen removal. The capacity for substantial penile erection is retained during copulation long after reflexive erections have diminished.


Year-to-year patterns of circulating levels of testosterone and corticosterone in relation to breeding density, experience, and reproductive success of the polygynous red-winged blackbird. To determine across-year patterns in plasma testosterone (T) and corticosterone (B) levels in free-living birds, we took blood samples in the same 2-week breeding period during 4 consecutive years from territorial male red-winged blackbirds. We used our data to test predictions of the "challenge hypothesis" of T secretion and also to examine hormonal correlations with age, breeding experience, breeding density, and reproductive success. Average T and B levels across years were not significantly different. Within individuals, T levels between years were, in general, highly variable. T levels of males with territories in high-density breeding areas were significantly higher than those of males with territories in low-density areas. T levels were positively correlated with harem size and there was a trend for males with high T levels to fledge more offspring. We found no relationships between T and B levels and male age or breeding experience. Some results of the study were consistent with the predictions of the challenge hypothesis. This study constitutes one of the few examinations of across-year patterns in circulating hormone levels in wild populations.


Prenatal diethylstilbestrol exposure and performance on college entrance examinations. The fetal rodent brain is permanently altered by exposure to sex hormones. Long-term effects of prenatal sex hormones on the human brain are far less clear. In order to explore such effects, we studied a measure of cognitive function among young adults who had been exposed in utero to a powerful synthetic estrogen. In a randomized clinical trial conducted at the University of Chicago in 1950-1952, 1646 pregnant women were randomly assigned to receive either high doses of diethylstilbestrol or placebo. Women in this study gave birth to 1653 liveborn infants, of whom 1603 (820 sons and 783 daughters) survived to their fifteenth birthday. College entrance examination scores were obtained for 42% of these offspring. No differences in test performance were seen among exposed daughters. Among sons, test scores were marginally higher among the exposed, probably due to chance.


Management of early postdischarge adjustment reactions following psychiatric hospitalization. Psychiatric patients frequently experience serious symptoms and demonstrate disturbed behaviors in the very early postdischarge period. Based on 25 years of clinical experience, the author reviews symptoms and behaviors that can occur and notes that they should most often be viewed as adjustment reactions rather than as exacerbations of the primary illness. A team approach to management of early postdischarge reactions that uses a psychiatrist and psychiatric nurse is effective. Interventions include forewarning inpatients that problems may occur and helping them identify potential problems. Social skills training, learning therapies, and family counseling help patients prepare to cope. Accompanying patients home on passes during hospitalization is helpful, as is inviting them to visit the hospital during their first days at home. Scheduling initial office visits within three days of discharge is another way of easing a difficult transition.


Experimental comparison of the effects of three treatment programs for homeless mentally ill people. A longitudinal experimental design was used to compare the effectiveness of three community-based treatment programs serving homeless mentally ill people: traditional outpatient treatment offered by a mental health clinic, a daytime drop-in center, and a continuous treatment team program that included assertive outreach, a high staff-to-client ratio, and intensive case management. At 12-month follow-up, clients in all three treatment programs spent fewer days per month homeless, showed fewer psychiatric symptoms, and had increased income, interpersonal adjustment, and self-esteem. Clients in the continuous treatment program had more contact with their treatment program, were more satisfied with their program, spent fewer days homeless, and used more community services and resources than clients in the other two programs.


Posttraumatic stress disorder among homeless men and women. Six hundred homeless men and 300 homeless women in St. Louis were systematically interviewed using the revised Diagnostic Interview Schedule that includes a module for assessment of posttraumatic stress disorder (PTSD). Most subjects with PTSD had an additional life-time psychiatric diagnosis. No consistent pattern of association was apparent, however, between individual diagnoses and either traumatic events or PTSD. In almost three-fourths of both men and women, the onset of PTSD had preceded the onset of homelessness. Childhood histories of abuse and family fighting were predictive of both traumatic events and PTSD. The results suggest that factors leading to PTSD in the study sample began long before the onset of homelessness and may overlap with factors operative in the genesis of homelessness.


Homeless adults without apparent medical and psychiatric impairment: onset of morbidity over time. A cross-sectional study compared characteristics of homeless adults with and without substance abuse, physical health problems, and history of psychiatric hospitalization when they first became homeless. Self-report data on demographic characteristics, adverse events in childhood, and history of medical disorders were collected from 1,399 homeless adults who used three shelters in Santa Clara County, California, during a five-month winter period in 1989 and 1990 (96 percent response rate). A total of 45.6 percent of the respondents reported no impairments when they first became homeless. They were distinguished from those with impairments at onset of homelessness by their younger age, minority status, lower educational attainment, and lower frequency of adverse events in childhood. Respondents who reported no impairments when they first became homeless were likely to develop addictive and psychiatric disorders over time. Those who had been homeless five years or more reported high rates of alcohol abuse (34.5 percent), illegal drug use (24.1 percent), and psychiatric hospitalization (20.7 percent).


Privilege and discharge decisions for psychiatric inpatients with dysphagia. Psychiatric patients have an increased risk for choking compared with the general population because of risk factors such as medication side effects and food gorging. A state hospital program for managing patients with dysphagia, or difficulty swallowing, includes interventions such as modified diets, mealtime monitoring, and adjusting psychotropic medications. Clinicians may find it difficult to make decisions about privileges and placement for dysphagic patients who do not comply with dietary modifications in unsupervised settings. For many such patients, close supervision and even placement on a locked ward may seem necessary. The authors recommend a risk-benefit approach: clinicians must balance the safety afforded by restrictions against the benefits of increased privileges or placement in a less restrictive setting. Quality of life and patients' preferences must also be considered.


Alcoholism in older persons: a review of the literature. Alcohol abuse and dependence in elderly persons is of growing social concern. The most consistent findings of cross-sectional and longitudinal studies are that the quantity and frequency of alcohol consumption is higher in elderly men than in elderly women, as is the prevalence of alcohol-related problems. Most studies show a decrease with age in consumption and alcohol-related problems among heavy drinkers. Longitudinal studies show no changes in consumption among light drinkers. Elderly persons with lower incomes consume less alcohol than those with higher incomes. Hospitalized and outpatient populations have more problem drinkers, and the elderly alcoholic is at greater risk for medical and psychiatric comorbidity. About one-third to one-half of elderly alcoholics experience the onset of problem drinking in middle or late life. Outcomes seem to be better for those who have late-onset drinking and may be improved for those treated in same-age rather than mixed-age groups.


State-university collaboration to enhance public psychiatric services in western Pennsylvania. A collaboration established in 1974 between the Pennsylvania State Office of Mental Health and Western Psychiatric Institute and Clinic of the University of Pittsburgh Medical Center provides a comprehensive program of continuing mental health education, skills development, and program consultation to state hospitals and publicly funded community programs in rural and semirural western Pennsylvania. The authors describe the development of the program and discuss its current organization and activities. In 1990-91 a total of 60 faculty and 150 staff members from the institute contributed more than 1,200 hours of direct programming. The collaboration's activities have broadened in both hospital and community sectors and currently involve all of the university's health and medical care schools.


State health reform. Five trends that will transform hospitals. As states struggle to forge health care reform plans that meet the opposing objectives of cutting costs and improving access, one thing is certain: Their initiatives will create profound changes to health care and to hospital operations. In fact, five factors emerging from the state reform debate could drastically transform hospitals: reregulation, explosive managed care growth, uncertain state funding mechanisms, new insurance measures, and greater hospital accountability for cost and quality. "This is a wake-up call for hospitals to get very realistic about the cost concerns that are out there," says one hospital executive. Especially since these state efforts are a precursor to national reform.


The new finance department: CQI triggers big changes in role. As hospitals implement total quality management and CQI programs and experiment with patient-centered care initiatives, financial officers are moving away from their traditional "bean-counter" role toward a broader role as part of a leadership team crafting a vision of the future.


Making mandatory medical staff meetings more physician-friendly. In an effort to improve hospital-medical staff relations, a number of hospitals are making changes in their medical staff meetings structures--putting time limits on meetings, modifying attendance requirements, even timing comments. One California hospital even uses an egg timer to facilitate the monthly meetings of its medical staff executive committee.


Work redesign calls for new pay and performance plans. TQM/CQI and patient-centered care initiatives are also spurring changes in hospitals' employee payment and performance systems. The tricky part is that those systems have to change at the same time delivery systems are changing. "We feel we can't first develop new care delivery systems and then spend the next two years adjusting compensation systems," says one executive.


JCAHO introduces three new areas of survey concentration. The Joint Commission has released a new and improved 1993 Accreditation Manual for Hospitals. New to the manual are three chapters of standards that will be used to survey areas that include patient and family education and staff training and orientation. The manual is also shorter.


Hospitals and medical staffs: the concept of planning takes on new meaning. Broad trends in health care are redefining medical staff planning. Hospital CEOs are recognizing the critical need to involve their physicians in hospital strategic planning at many levels. Gone are the days when it was sufficient to invite medical staff members to annual planning retreats and add individual physicians to boards; hospitals that thrive in the 1990s will be those that have created strong strategic links with their physicians. At the same time, medical staff development planning is changing in important ways. Recent federal government alerts on fraud and abuse and inurement in physician-recruiting activities are leading hospitals to document community benefit in their recruitment efforts. And hospital executives now realize that changes in the physician market will require them to plan carefully in order to ensure a strong base of primary care and other much-needed physicians. These two trends present CEOs with multilayered challenges. Following are reports on what leading-edge hospitals are doing in both areas.


Hospitals find ways to integrate risk-management functions. Hospitals that have successfully integrated the risk-management function with other hospital departments say that the process requires both education and teamwork. Among the major challenges: overcoming physician reluctance to participate in risk-management activities, and linking risk management to quality assurance.


Rochester: community rating = insurance access. Despite opposition from some quarters, the New York State Legislature recently passed a small-group reform bill requiring community rating in health insurance throughout that state. The health care system of Rochester, NY, the author contends, proves that community rating is both good public policy and good business practice.


Polymorphism and differential selection for the sexes. Various genetic models with different fitnesses for the sexes are investigated. Only a limited set of fitness values will result in a stable polymorphism, and the rate of approach to these equilibrium frequencies is extremely slow. These results indicate that there are problems associated with the interpretation of some human genetic traits by such models.


Inbreeding effects on reproductive outcome: a study based on a large sample from the endogamous Vadde of Kolleru Lake, Andhra Pradesh, India. Inbreeding effects on reproductive outcome vis-à-vis fertility, prenatal loss and prereproductive mortality, and secondary sex ratio of live and dead children were examined in a large sample of 2078 women of the Vadde fishing population of Kolleru Lake in Andhra Pradesh, India. Demographically, this population is a single endogamous unit. By using an exponential regression model with the proportion of offspring survival as a dependent variable and the inbreeding coefficient as an independent variable, I further examined the inbreeding effects. The results were compared with results from other fishing groups and other southern Indian and non-Indian populations. The results among the Vadde were consistent with those found for other groups of Telugu-speaking fishermen and several other southern Indian populations in that the effects were neither perceptible nor significant. The average B value and the number of lethal equivalents found for the highly inbred southern Indian populations in general and for the Vadde in particular were much smaller than those from other parts of the world, providing empirical support to Sanghvi's hypothesis on long-term effects of inbreeding.


Digital dermatoglyphic patterns of Eskimo and Amerindian populations: relationships between geographic, dermatoglyphic, genetic, and linguistic distances. Dermatoglyphic traits have been used to assess population affinities and structure. Here, we describe the digital patterns of four Eskimo populations from Alaska: two Yupik-speaking villages from St. Lawrence Island and two Inupik groups presently residing on mainland Alaska. For a broader evolutionary perspective, these four Eskimo populations are compared to other Inuit groups, to North American Indian populations, and to Siberian aggregates. The genetic structures of 18 New and Old World populations were explored using R-matrix plots and Wright's FST values. The relationships between dermatoglyphic, blood genetic, geographic, and linguistic distances were assessed by comparing matrices through Mantel correlations and through partial and multiple correlations. Statistically significant relationships between dermatoglyphics and genetics, genetics and geography, and geography and language were revealed. In addition, significant correlations between dermatoglyphics and geography, with linguistic variation constant, were noted for females but not for males. These results attest to the usefulness of dermatoglyphics in resolving various evolutionary questions concerning normal human variation.


Genetic and dermatoglyphic distances among Basque Valleys. Palmar dermatoglyphics of a sample including 552 males and 701 females from 8 Basque valleys were analyzed. We studied the frequency of palmar pattern types and compared them using correspondence the frequency of palmar pattern types and compared them using correspondence analysis. The results of this comparative study show that there is diversity among valleys and also that this diversity depends on the trait and on sex. Genetic drift could explain this variability found in the Basque population.


Nonmetric tooth crown traits in the Ami tribe, Taiwan aborigines: comparisons with other east Asian populations. The frequencies of occurrence of 17 tooth crown traits in the living Ami tribe, which inhabits the east coast of Taiwan, were investigated and compared with other East Asian populations based on Turner's (1987) Mongoloid dental variation theory. Principal coordinate analysis based on Smith's mean measure of divergence using frequencies of the 17 traits suggests that the Ami tribe together with the Yami tribe and the Bunun tribe is included in the sinodont group typical of the Chinese mainland and northeast Asia. In light of these results and the estimated distribution of sinodonty and sundadonty in the past and the present, we speculate that the gene flow from Chinese mainlanders to native sundadonts, who seem to have migrated northward to Taiwan, contributed significantly to the formation of the living Taiwan aboriginal groups, sinodonts. Among the aboriginal tribes of Taiwan, the Ami have characteristics intermediate between those of the Yami and the Bunun. The relative positions of these tribes in East Asian populations suggests that the extent of sinodontification and of genetic isolation is one of the causes of the intertribal variation.


Rate of growth in length of Bangladeshi infants as a function of attained length. We examine variation in the rate of growth in length of breast-feeding infants from rural Bangladesh. These data were collected between November 1985 and February 1986 from two rural sites. Eighty-eight infants, ranging from birth to 4 months of age at the start of the study and their mothers were measured monthly for 4 months. Length increased linearly with age over this 4-month period (infants' average bias-adjusted R2 = 0.90). The relationship between infant rate of growth in length and attained length was analyzed by two different methods: Oldham's (1962) method of regressing rate of growth on mean length and Blomqvist's (1977) method of regressing rate of growth on estimated initial length. The methods gave similar results. The rate of growth was negatively associated with mean infant length over the 4-month period (p less than 0.001); that is, shorter infants grew at a faster rate than longer infants. For every centimeter shorter the infant was, the rate of growth was 0.1 cm/mo faster on average; the effect was greater among males than among females. The average rate of growth was greater for males than for females and greater in financially solvent households and varied by site. Infant growth rate was slower among older infants than among younger infants, as expected. However, after adjusting for mean infant length, age was no longer significantly associated with infant growth rate, although mean infant length remained highly significant. Forty-one percent of the variation in infant rate of growth in length was explained by mean infant length, sex, sex by length interaction, household financial solvency, and site.


Short-term projections of AIDS cases in Mexico. We attempt to analyze and predict the behavior of the AIDS epidemic in Mexico. The reporting delay is corrected by using a cluster analysis, and the corrected data are used to make short-term projections by extrapolation, by fitting linear and log-linear models, and by back-calculation. The incubation period is assumed to have a Weibull distribution, and step functions are used for the infection functions. Most of the methods predict a mean of 25,000 accumulated cases by the end of 1993, and a comparison of the predictions with actual data up to November 1990 shows good agreement in all cases except the log-transformation linear model. The data for 1990 also show the reporting delay correction to be adequate in most cases.


Y-chromosome-specific haplotypes of Jews detected by probes 49f and 49a. A sample of Ashkenazic and Sephardic Jews has been studied with respect to haplotypes at the 49f-49a Y-specific DNA probes. Only seven haplotypes were found in Jews, three of them (VII, VIII, and XI) being the most widespread. Haplotype distribution in the European non-Jewish population is different.


[Homology study of leptospires by molecular hybridization]. Nick translation and random primer labelling method were applied to prepare three genomic DNA probes from Leptospira interrogans strain 017, Leptospira biflexa strain Patoc I and Leptonema illini strain 3055, and then hybridized with DNA of 17 strains leptospires from different genus, species, serogroup and serovar. The results showed no homology between Leptospira and Leptonema, and only a low degree of homology between L. interrogans and L. biflexa but it showed a high degree of homology among L. interrogans. The study also proved the possibility to establish a DNA probe prepared from a single leptospira strain to detect different serovars.


[A mid-infrared spectra study of a series of benzocrown ether and macrocyclic polyether diester]. With a Nicolet 20 SX B Fourier Transform infrared spectrometer, the mid-infrared spectra in region of 4000-400 cm-1 were observed for a series of benzocrown ether and macrocyclic polyether diester. A difference in the wavelength of absorption of the ether bond was found between benzocrown ether and macrocyclic polyether diester. The wavelength of absorption of the ether bond of the compounds I-IV moved down about 25 cm-1, as compared with that of the compounds V-IX.


[Solid phase clean-up and determination of vesnarinone in plasma by high performance liquid chromatography]. Vesnarinone, a new positive inotropic agent, was synthesized by Tominaga et al of the Otsuka Pharmaceutical Co. Ltd. Institute in 1982. Determination of Vesnarinone plasma level in dogs by reversed-phase HPLC was developed and presented in this paper. A plasma sample preparation was carried out by solid phase cleaned-up through neutral aluminum column (10 x 1 cm I.D.) eluting with methanol. The chromatography consisted of a slim-pack CLC ODS column (150 x 6.0 mm I.D.) with methanol: 1 mmol/L HAc (55:45) as the mobile phase at a flow rate of 0.8 ml/min. Spectrophotometric detection was at 271 nm, and column temperature was at 25 degrees C. Using 3,4-dihydro-6-(4-(4-methoxybenzoyl)-1-piperazinyl)-2 (1H) quinolinone as an internal standard. The cleaned-up procedure is simple, rapid and satisfactory. The detection limit of vesnarinone was 0.5 ng/ml (R/N, 3:1). The calibration curve was liner (r = 0.99998) in the concentration range of 5-500 ng/10 microliters. Within-day and day-to-day precisions (CV%) were 1.56 and 1.98. The recoveries obtained from cleaned-up and spiked plasma samples were up to 86.52 +/- 2.77% and 96.26 +/- 5.46%, respectively.


[Preparation of a new tissue-based biosensor]. A new adenosine electrode using rabbit thymus tissue as catalyst is described. The tissue membrane was immobilized at the surface of an ammonia gassensing electrode for determination of adenosine in body fluid or tissue samples. This tissue-based biosensor showed excellent selectivity, and sensitivity, with a response slope of 51.2mV/decade, linear range of 3.16 x 10(-5)-5.62 x 10(-3) mol/L, coefficients of variation ranging from 1% to 5.72% (n = 7, m = 5), recoveries of adenosine in rabbit blood from 94.9% to 108.4% (n = 5, m = 5), and response time of 7 min. The life-time of the electrode was 25 days and 14 analogous compounds showed no significant interference.


[Plasmid analysis and antibiotic resistance pattern measurement of Shigella in Chengdu]. A total of 81 strains of Shigella isolated from sporadic cases and cases in an outbreak of Shigella sonnei infection was analysed by plasmid profiles and antibiotic resistance patterns. The strains had a high resistance patterns. The strains had a high resistance of ampicillin, tetracycline, trimethoprim-sulfamethoxazole and chloramphenicol; while 97.2% of the strains were susceptible to gentamicin, kanamycin. The results showed that 50% of the strains harbored plasmid patterns within each species, which indicated that many genetically different strains of Shigella were responsible for dysentery epidemics in Chengdu. According to the results of the plasmid analysis and antibiotic resistance pattern measurement made in the outbreak strains, it was clearly seen that most of the strains were similar or fundamentally similar, which suggested that one or two strains with genetical homology were the main causative agent for the outbreak. We think that plasmid profiles and antibiotic resistance patterns are applicable to the identification of the outbreak strains of dysentery as an efficient epidemiologic tool.


[Biochemical studies of the temporomandibular ligament in dogs]. The purpose of this investigation was to determine the biochemical composition of the normal adult dog's temporomandibular ligament (TML) along its ligament (TML) along its length. Water content and hexosamine concentration as an index of glycosaminoglycan content and collagen concentration were quantified; and the results compared statistically with anatomically identified TML segments. The TML contained lower water content and higher collagen, hexosamine concentration than capsule (P less than 0.05). All the contents of these components in different locations along the ligament were not significantly different, with the exception that on the insertion site of the temporal eminence the concentration of the hexosamine was less than that in other areas (P less than 0.05). This study provided useful data for the analysis of TML functions and its mechanical properties.


[Technique of measuring esophageal pressure in rabbits and its application to measurement of pulmonary compliance and resistance]. Esophageal pressure (Pes) was measured under non-invasive condition by an esophageal balloon made by ourselves, and the optimal position of rabbits for measuring Pes was explored. The correspondence between Pes and pleural pressure (Ppl) was examined in rabbits during different depths of breathing with varied lengths of balloon and varied depths of insertion of balloon into the esophagus. The results showed that the amplitude of Pes only on supine position was higher than that on the left and right lateral and prone positions, but no significant difference was noticed among the 3 positions. The optimal balloon length was 4-6 cm. The optimal depth of insertion of balloon into the esophagus was 20 cm. The pulmonary dynamic compliance and pulmonary resistance were measured in 11 white rabbits with 5 cm balloon, 20 cm depth of insertion, on the left lateral position. The results were correspondent with other authors. No significant difference was noticed in repeated measurements on 8 of the 11 rabbits 5 days later.