Detection and partial purification of a potent mitogenic factor for human thyroid follicular cells.

Normal adult human thyroid follicular cells have an extremely limited proliferative capacity in vitro. No previously studied mitogen, including thyrotropin (TSH) or epidermal growth factor (EGF), has in our hands resulted in a significant improvement over the 3-4% nuclear [3H]thymidine pulse-labelling index (LI) obtainable with 10% fetal calf serum. Here we report the detection in the conditioned medium from a sub-clone of NIH3T3 fibroblasts of a mitogenic activity capable of increasing this response up to 10-fold, to an LI of over 20%, together with an even greater relative stimulation of mitotic activity. Preliminary characterisation has excluded EGF and TGF alpha, and demonstrated that the activity is bound reversibly by heparin-Sepharose, thus pointing to a member of the heparin-binding fibroblast- or hepatocyte-growth factor families. This material should have wide practical application in facilitating primary culture of follicular cells, and may reveal new mechanisms of stromal-epithelial interaction regulating normal and neoplastic thyroid growth in vivo.     

The minimum concentration of fibrinogen needed for platelet aggregation using ADP.

OBJECTIVE: Determine the minimum concentration of plasma fibrinogen needed to stimulate the aggregation of platelets, collected from normal subjects, using ADP. DESIGN: Platelet rich plasmas (300 x 10(9) platelets/L) were made and adjusted to final fibrinogen concentrations of 75, 19, 5, and 0 mg/dL using fibrinogen free serum. Each fibrinogen concentration in all twelve subjects was aggregated with ADP SETTING: Research laboratory in the Department of Clinical Laboratory Science at Saint Louis University. PARTICIPANTS: Twelve healthy volunteers of both genders, between the ages of 18 and 60 years who were not pregnant and weighed at least 110 pounds were included in the study. Subjects were excluded from the study if they had ingested aspirin within one week prior to blood collection. In addition, subjects with a history of bleeding disorders such as afibrinogenemia, hypofibrinogenemia, von Willebrand disease, and Bernand-Soulier disease were rejected from the study. MAIN OUTCOME MEASURES: Platelet aggregation tracings were analyzed for amplitude and compared across plasma fibrinogen concentrations. In addition, the type of curve (monophasic vs. biphasic), smoothness and aggregation stability were also noted. RESULTS: The results show that aggregation occurred with every dilution of fibrinogen tested and that the amplitude of the aggregation curves appears not to be dependent on plasma fibrinogen. CONCLUSIONS: The results indicate that platelets from healthy individuals previously exposed to normal fibrinogen levels will aggregate equally well in decreasing plasma fibrinogen concentrations and even in the absence of plasma fibrinogen using ADP as the aggregator.     

Ovarian actinomycosis presenting as acute peritonitis.

A patient with acute peritonitis due to ovarian actinomycosis and the association of this disease with the intrauterine contraceptive device is discussed. Because adequate treatment requires prolonged antibiotics the diagnosis should be sure. Unfortunately if antibiotics are given pre-operatively culture is usually unsuccessful and thus careful examination of all removed material is essential. As many of these women are of child bearing age surgical intervention should aim to preserve fertility wherever possible.     

Cloning and expression of SLC1OA4,a putative organic anion transport protein

AIM:To determine if novel bile acid transporters may be expressed in human tissues.METHODS:SLC10A1 (NTCP) was used as a probe to search the NCBI database for homology to previously uncharacterized ESTs. The homology search identified an EST (termed SLC10A4) that shares sequence identity with SLC10A1 and SLC10A2 (ASBT). We performed Northern blot analysis and RT-PCR to determine the tissue distribution of SLC10A4. SLC10A4 was cloned in frame with an epitope tag and overexpressed in CHO cells to determine cellular localization and functional analysis of bile acid uptake.RESULTS:Northern analysis revealed that SLC 10A4 mRNA is ubiquitously expressed fn human tissues with the highest levels of mRNA expression in brain,placenta, and liver. In SLC10A4-transfected CHO cells,immunoblotting analysis and immunofluorescence staining demonstrated a 49-kDa protein that is expressed at the plasma membrane and intracellular compartments.Functional analysis of SLC10A4 showed no significant taurocholate uptake in the presence of sodium when compared to untransfected CHO cells.CONCLUSION:To date, we have shown that this protein has no capacity to transport taurocholate relative to SLC1041; however, given its ubiquitous tissue distribution, it may play a more active role in transporting other endogenous organic anions.     

Complications of the cardiopulmonary stress test in patients with depressed left ventricular systolic function.

BACKGROUND: The exercise test has a recognized lower risk of complications when used in the general population and in coronary artery diseased patients, but from a theoretical point of view should have a higher rate of complications when performed in patients with chronic heart failure (CHF). AIMS: To characterize and assess the type and incidence of complications during cardiopulmonary stress test (CPX) in patients with depressed left ventricular systolic function in comparison with a group of patients and individuals with normal function. METHODS: Retrospective analysis of the 334 consecutive CPX performed for risk stratification in 198 patients with a left ventricular ejection fraction below 40% (Group A) and 180 consecutive CPX performed in 78 subjects with normal function (Group B). The two groups were compared with respect to demographic data, CPX parameters and specific complications. Results: Major complications during the tests occurred only in 14 tests of Group A (4.2%, p = 0.012). Non-sustained ventricular tachycardia, <6 beats, occurred in 7 group A and 2 group B tests. The absence of coronary artery disease was the only independent predictor for complications. CONCLUSIONS: Major CPX complications occurred only in patients with impaired left ventricular systolic function. Heart failure patients showed a low probability (around 4%) for complications during CPX, significantly higher and more severe than the risk in the group of patients with normal ventricular function, allowing us to recommend that CPX in patients with heart failure should be performed in a hospital setting under the supervision of a physician with specific training.     

乙型肝炎表面抗原“免疫逃避”突变体在哺乳动物细胞中的表达

利用乙型肝炎病毒ＤＮＡ开放框架上的ＢａｍＨＩ和ＨｐａＩ位点，酶切消化质粒载体ＰＥｃｏｂ６（含双拷贝ＨＢＶＤＮＡ），得到约９００ｂｐ的ＨＢＶ－Ｓ基因片断。将其插入到噬菌粒载体ＰＢｌｕｅｓｃｒｉｐｔｓｋ＋的ＳｍａＩ位点上。然后通过体外寡核苷酸介导的人工定点突变获得一系列（共１２种）Ｓ基因“免疫逃避”突变型。再通过ＥＢ病毒真核表达载体ｐＭＥＰ４上的ＢａｍＨＩ和Ｋｐｎｌ位点将噬菌粒ｐＢｌｕｅｓｃｒｉｐｓｋ＋上的Ｓ基因突变型片断定向克隆到ＰＭＥＰ４上，从而构建了含乙肝Ｓ基因突变型的重组质粒ｐＭＥＰ４ＨＢＳＭ。用其转染人肝癌传代细胞系ＨｅｐＧ２，经潮霉素选择，三周后获得抗性细胞克隆。经用抗ＨＢｓ单克隆抗体（含针对ＨＢｓＡｇ“ａ”抗原决定簇）检测除含变异体１４５（即１４５位上甘氨酸为精氨酸替代）外其余抗变异体ＨＢｓＡｇ均为阳性。经Ｗｅｓｔｅｒｎｂｌｏｔ证实变异体１４５，在分子量约为２３ＫＤ处有一特异ＨＢｓＡｇ蛋白带。     

Bioavailability of iron in prenatal multivitamin/multimineral supplements administered to pregnant teenagers

Absorption of elemental iron from three single-daily-dose prenatal multivitamin/multimineral supplements was compared in bioavailability studies of subjects under fasting and postprandial conditions. Each of the supplements (Stuartnatal 1 + 1, Stuart Prenatal, and Materna 1.60) contained 60 or 65 mg of iron. The subjects, teenagers in the second trimester of pregnancy, were assigned to the fasting or postprandial conditions at intervals of three to seven days. For the postprandial condition, subjects took the supplement after eating a standardized meal that had been designed to provide a minimal amount of iron and no caffeine (which inhibits iron absorption); blood was drawn at 0, 1, 3, 6, and 8 hours. Bioavailability studies showed that all three supplements provided adequate levels (at least 3.5 mg) of absorbed iron when taken postprandially. The highest quantity of net iron transport as well as the most rapid intestinal transport in either condition was yielded by Stuartnatal 1 + 1 taken in the fasting state. In the postprandial condition, Materna 1.60 demonstrated the fastest transport time and highest net iron transport; when given to a subject in the fasting condition, however, this supplement provided less than the recommended 3.5 mg of absorbed iron. These results suggest that iron absorption is influenced by supplement formulation as well as the presence or absence of food. Considering variations in compliance when iron is to be taken on an empty stomach as well as variable absorption in the fasting state, the most reliable supplementation would appear to be achieved by instructing pregnant women to take multivitamin/multimineral supplements at mealtime.     

Comparison of questionnaire screening and urine toxicology for detection of pregnancy complicated by substance use.

OBJECTIVES: To compare the efficacy of structured questionnaire screening and prenatal urine toxicology for the detection of substance use by pregnant women, and to describe substance use patterns in a group of women presenting to a university-based obstetric clinic. METHODS: All patients presenting to our obstetric clinic for their first prenatal visit were evaluated for evidence of current use of alcohol or any illicit substances. Nursing personnel administered an extensive questionnaire, which included detailed questions about past and current substance use patterns. Urine samples were examined by a commercial laboratory for alcohol and a number of illicit substances. Current users were defined as those who admitted use within the past 30 days or who had positive urine toxicology. RESULTS: Fifty-eight of 302 patients (19.2%) were identified as current users of alcohol or illicit substances. Only 17 of 41 women (41.5%) with toxicologic evidence of recent use admitted to current use. Only 17 of 34 (50.0%) admitting to current use had toxicology positive for any substance. Patients with a history of multiple past substance use were significantly more likely to have positive urine toxicology than those without such a history (26.1 versus 7.4%; P < .005). Among current users, multiple substance use (34.5%) and cigarette smoking (52%) were common. CONCLUSION: A screening combination of structured questionnaire and universal urine toxicology identifies more current users than either technique alone, and neither is clearly superior to the other. A history of multiple substance use may be an important indicator of current use.