The number and location of glycans on influenza hemagglutinin determine folding and association with calnexin and calreticulin

Large quantities of recombinant acid alpha-glucosidase are needed for in vivo experimentation of enzyme replacement therapy in Pompe disease. We describe a new purification method for the purification of this recombinant enzyme from tissue culture medium consisting of concanavalin A affinity chromatography, hydrophobic interaction chromatography, affinity chromatography on Superdex, and anion exchange chromatography. The new method is amenable to scale up, and has increased speed, and improved reproducibility with similar high yield and purification efficiency when compared to previous methods.     

The CMRF-35 mAb recognizes a second leukocyte membrane molecule with a domain similar to the poly Ig receptor

In order to understand the problems of undesired infertility in the Gambia, where the desire for children and fertility is very high, a population based estimate of the frequency of sub-/infertility was undertaken. A survey was used in a representative random sample of the population. The study included an assessment of health care available for infertile couples in the different types and levels of the formal and traditional health system. Primary sterility was found to be fairly uncommon (3%), and secondary infertility to be more frequent (6%). Half of the infertile couples failed to seek formal health care, and they had to reach a certain level of care in order to be properly managed. As investigations are very basic and treatment possibilities scarce, many forms of alternative care are often sought. In addressing reproductive health in developing countries, a systematic primary health care approach to infertility in rural areas with limited resources should be developed.     

Spontaneous regression of renal cell carcinoma: a pitfall in diagnosis of renal lesions

Lobar atrophy is a rare morphologic change of the liver. We describe a 73-year-old woman with mild liver dysfunction and history of Sj?gren's syndrome who had right hepatic lobar atrophy. Serum biochemistry levels were as follows: albumin, 4.5 g/dl; total bilirubin, 1.0 mg/dl; alanine aminotransferase, 25 international units/l; aspartate aminotransferase, 27 international units/l; alkaline phosphatase, 333 international units/l; and gamma-glutamyl transpeptidase, 332 international units/l. Serological data were as follows: rheumatoid factor, 27.9; anti-nuclear antibody, 1:640; and antismooth muscle antibody, 1:80. Viral markers for hepatitis B were all negative. Anti-hepatitis C virus (anti-c-100) was negative. Portal hypertension developed thereafter, and the patient died of hepatic failure at age 76. Postmortem examination revealed autoimmune hepatitis with moderate fibrosis, portal vein thrombus, and complete obstruction of the right hepatic duct due to hepatolithiasis. Terminal hepatic failure resulted from combination of decreased hepatic volume due to the right lobar atrophy, exacerbation of autoimmune hepatitis in the remnant left hepatic lobe, decreased portal venous blood flow due to thrombosis, portal hypertension, and cholangitis with hepatolithiasis. This is the first reported case of hepatic lobar atrophy due to autoimmune hepatitis. From a clinical standpoint, patients with hepatic lobar atrophy, even if asymptomatic, should be followed up with careful attention to progression of liver diseases, portal hypertension, and biliary complications.     

Alcohol choice and outcome expectancies in social drinkers

Eighteen male social drinkers underwent four training sessions during which they ingested two colour-coded drinks (red or blue, balanced for drink type); one containing alcohol (aliquots of 0.1 g/kg) and the other placebo (aliquots of orangeade). Following the training sessions, subjects were presented with both drinks, and instructed to choose the drink they felt like consuming and to indicate their preference for their chosen drink over the other drink. In addition, they were instructed to consume the first drink but that all subsequent drinks (total of six drinks), offered at 10-min intervals, were optional. A number of trait characteristics were assessed including alcohol outcome expectancies, drinking habits and personality traits. The acute effects of alcohol on mood was also evaluated by comparing subjective ratings following alcohol and placebo during the training sessions. Of the 18 subjects, 12 chose alcohol at least once ('samplers'), whereas six never chose alcohol ('non-samplers'). Over the three sessions, however, alcohol and placebo were chosen equally. When alcohol was chosen, subjects drank significantly more than when placebo was chosen, which may be consistent with a priming effect of drinking alcohol. The amount of alcohol drunk was seen to correlate with the alcohol expectancy factor 'sociability'. Subjective reports of feeling 'alert', 'clear-headed', 'quick-witted', and 'attentive' all showed a main effect of choosing behaviour (i.e. 'samplers'/'non-samplers'). Further analysis indicated that this effect was due to 'samplers' reporting increased subjective ratings of these mood states following the ingestion of alcohol compared to 'non-samplers'. These increased subjective ratings were also positively correlated with the amount of alcohol consumed by the subjects during the choice procedure. No other relationships were found between the amount of alcohol consumed and any of the other state or trait measures. These data suggest that social drinkers who sample alcohol in a laboratory setting can be primed by alcohol to consume more. The results also indicated that the amount drunk was related to the degree to which subjects expected alcohol to increase sociability and to reports of subjective stimulant effects of alcohol (e.g., 'alert', 'clear-headed', 'quick-witted', and 'attentive').     

Modulation of intestinal estrogen receptor by ovariectomy, estrogen and growth hormone

Ovarian hormone deficiency decreases and estrogen (E2) and growth hormone (GH) administrations increase intestinal absorption of calcium (Ca++). However, the underlying mechanisms are uncertain. To examine whether alterations in the binding characteristics of intestinal estrogen receptors (ERs) are involved, we developed and validated methods for simultaneous measurement of intestinal ERs in cytosolic and nuclear fractions and applied these techniques to four groups of female rats: sham-operated, ovariectomized (Ovx), Ovx + 5 micrograms E2/kg b.wt./day and Ovx + 8 mg GH/kg. b.wt./day. All animals were killed on day 21, and mucosal cells harvested from the duodenum for ER determination. The cytosolic and nuclear ERs were 117.2 +/- 2.7 fmol/mg protein and 64.9 +/- 1.2 fmol/mg DNA, respectively, in sham-operated rats and decreased by 16.1% and 17.0% to 98.4 +/- 1.7 fmol/mg protein and 53.8 +/- 1.3 fmol/mg DNA, respectively in Ovx rats (P < .001). E2 therapy prevented completely the decrease in cytosolic and nuclear ERs that occurred in Ovx rat (126.1 +/- 2.9 fmol/mg protein and 68.0 +/- 3.0 fmol/mg DNA, respectively, in the E2-treated group). Similarly, GH administration prevented the decrease in cytosolic and nuclear ERs that resulted from ovariectomy (119.2 +/- 3.2 fmol/mg protein and 63.4 +/- 1.3 fmol/mg DNA, respectively, in the GH-treated group). The Kd of nuclear ER-ligand complex was 2.0 +/- 0.03 nM in sham-operated rats and was slightly modulated by Ovx, E2 and GH (3.3 +/- 0.02, 2.33 +/- 0.09 and 2.23 +/- 0.04 nM, respectively, P < .001), but the Kd of cytosolic ER-ligand complex was not altered by Ovx, E2 or GH. Our findings indicate that E2 deficiency down-regulates, whereas E2 and GH administrations up-regulate intestinal ERs and prevent ovariectomy-induced decrease in receptor binding affinity. We conclude that E2 deficiency, E2 and GH may modulate intestinal Ca++ absorption, in part, by altering the abundance and binding characteristics of intestinal ERs.     

Levels of urinary beta-core fragment, total oestriol, and the ratio of the two in second-trimester screening for Down syndrome

Levels of beta-core fragment and total oestriol in second-trimester maternal urine samples were measured in 32 Down syndrome pregnancies and 206 control pregnancies. Beta-core fragment and total oestriol values were corrected for the urinary creatinine level and expressed as multiples of the control medians (MOM). In addition, the ratio of the beta-core fragment level to the total oestriol level, without creatinine correction, was calculated, and expressed as MOM values. The median beta-core fragment, total oestriol, and ratio levels in Down syndrome cases were 5.42, 0.64, and 9.32 MOM, respectively. In the Down syndrome pregnancies, 66 per cent of the beta-core fragment levels were above the 95th centile of control levels, while 22 per cent of the total oestriol levels were below the fifth centile of control levels. In combination with maternal age, measurement of beta-core fragment and total oestriol levels in Down syndrome pregnancy resulted in an 80 per cent detection rate at a 5 per cent false-positive rate. Use of the ratio resulted in a univariate detection rate of 72 per cent. In combination with maternal age, the ratio resulted in a detection rate of 81 per cent at a 5 per cent false-positive rate. Based on this unmatched study, the measurement of a ratio of beta-core fragment to total oestriol levels, without the need for creatinine correction, may be useful in screening for fetal Down syndrome in second-trimester urine.     

Cell-permeable ceramides prevent the activation of phospholipase D by ADP-ribosylation factor and RhoA

The mechanism of inhibition of phospholipase D (PLD) by ceramides was determined using granulocytes differentiated from human promyelocytic leukemic (HL-60) cells. In a cell-free system, hydrolysis of phosphatidylcholine by membrane-bound PLD depended upon phosphatidylinositol 4,5-bisphosphate, guanosine 5'-3-O-(thio)triphosphate) (GTPgammaS), and cytosolic factors including ADP-ribosylating factor (ARF) and RhoA. C2-(N-acetyl-), C8- (N-octanoyl-), and long-chain ceramides, but not dihydro-C2-ceramide, inhibited PLD activity. Apyrase or okadaic acid did not modify the inhibition of PLD by ceramides, indicating that the effect in the cell-free system was unlikely to be dependent upon a ceramide-stimulated kinase or phosphoprotein phosphatases. C2- and C8-ceramides prevented the GTPgammaS-induced translocation of ARF1 and RhoA from the cytosol to the membrane fraction. In whole cells, C2-ceramide, but not dihydro-C2-ceramide, inhibited the stimulation of PLD by N-formylmethionylleucylphenylalanine and decreased the amounts of ARF1, RhoA, CDC42, Rab4, and protein kinase C-alpha and -beta1 that were associated with the membrane fraction, but did not alter the distribution of protein kinase C-epsilon and -zeta. It is concluded that one mechanism by which ceramides prevent the activation of PLD is inhibition of the translocation to membranes of G-proteins and protein kinase C isoforms that are required for PLD activity.     

Spectrum of liver diseases in HIV infection

BACKGROUND: Most earlier reports on the spectrum of liver diseases in HIV-infected individuals originated from the West. OBJECTIVE: To study the spectrum of liver diseases in HIV-infected individuals. METHODS: Seventy four consecutive HIV-positive patients (57 men; age range 23-75 years, mean 34) were studied prospectively with clinical evaluation, liver function tests, ultrasonography, radioisotope liver scan, markers of hepatitis B (HBV) and C (HCV) viruses, and liver histology whenever necessary. RESULTS: Thirty four patients (45%) were chronic alcoholics. Mean (SD) absolute lymphocyte count was 2521 (1271)/mm3; count < 2000/ mm3 was present in 20 patients. Serum bilirubin, transaminases and alkaline phosphatase levels were elevated in 13%, 13% and 24% of patients, respectively. Ultrasonography detected an abscess in two patients (tuberculous-1, amebic-1). Evidence of exposure to HBV was present in 81% (HBsAg-12, hepatitis B core and/or surface antibody-48); anti-HCV antibody was positive in 29.7%. Five patients with liver tuberculosis (granuloma-4, abscess-1) had AFB either in liver tissue or lymph nodes. CONCLUSION: Chronic alcoholism, HBV and HCV infection, hepatic tuberculosis, and evidence of other liver disease were common in patients with HIV infection.