Rehydration methods to recover cysticercoids of the rat tapeworm Hymenolepis diminuta from dry flour beetle carcasses.

Terrestrial arthropods host a variety of helminth parasites, yet quantifying the intensity of infection in these hosts post-mortem is challenging because carcasses may desiccate quickly. We recovered cysticercoids of Hymenolepis diminuta from desiccated flour beetle (Tribolium confusum) carcasses by modifying a published insect rehydration procedure. Without rehydration, carcasses dissected more than 1 day post-mortem had noticeable degradation of cysticercoids. Mild rehydration (soaking in water only for 2 days, or 0.5-10% KOH for 1 h followed by 1 day in water, or 0.5% KOH for 1 day) left carcasses tough and time-consuming to dissect, but all parasites could be recovered and were similar in body size to fresh cysticercoids. Moderate rehydration (5-10% KOH for 1 day) allowed all parasites to be recovered and facilitated dissection by partially dissolving internal organs of the beetle while causing little degradation of the cysticercoids. Harsh rehydration (5-10% KOH for 1 day followed by 5 days in water) not only dissolved internal beetle tissues but also severely damaged cysticercoids, such that parasite counts were unreliable. The degree of initial carcass desiccation had little effect on results following rehydration. However, regardless of treatment used, intact cercomers were rarely retained on rehydrated cysticercoids. Rehydration was less successful on early developmental stages of the parasite, which were recovered reliably only as they neared the cysticercoid stage. This method has utility for studies of parasite-induced mortality by permitting accurate and reliable parasite counts from dead, desiccated hosts.

Impact of intra-abdominal fat on surgical outcome and overall survival of patients with gastric cancer.

BACKGROUND: The aim of this study was to evaluate the impact of obesity on surgical outcome and prognosis in patients with gastric cancer. METHODS: A total of 304 patients who underwent curative gastrectomy for gastric adenocarcinoma between January 2005 and March 2008were enrolled. Body mass index (BMI) was calculated before the operation and visceral fat area (VFA) was measured by abdominal computed tomography (CT). The patients were divided according to BMI class and VFA quartile. The influence of BMI and VFA on surgical outcome and survival was evaluated. RESULTS: The median BMI was 23.3 kg/m(2) and the median VFA was 103 cm(2). There was a significant positive correlation between BMI and VFA. According to BMI class and VFA quartile, there were no significant differences in patients' characteristics or surgical outcome, with the exception of a significantly longer operation time and fewer retrieved lymph nodes in patients with a high BMI and VFA. The unadjusted overall and disease free survival were not significantly different between BMI classes or VFA quartiles. CONCLUSIONS: Obesity, as represented by BMI and VFA, may not be a poor prognostic factor in patients with gastric cancer.

The role of small intestinal bacterial overgrowth in postgastrectomy patients.

BACKGROUND: Small intestinal bacterial overgrowth (SIBO) is expected in postgastrectomy patients; however, its role has not been clarified. This study was to estimate the prevalence of SIBO and investigate the clinical role of SIBO in postgastrectomy patients. METHODS: This prospective study involved 76 patients who underwent gastrectomy for early gastric cancer with no evidence of recurrence. An H(2)-CH(4) breath test with oral glucose challenge test was performed to diagnose SIBO and dumping syndrome. Sigstad dumping questionnaires, serum glucose, hematocrit and pulse rate were simultaneously monitored for every 30 min for 3 hours. KEY RESULTS: There were significant differences in SIBO between the postgastrectomy patients and controls (77.6%vs 6.7%, P < 0.01). Abdominal fullness or borborygmus during oral glucose load were more common in SIBO-positive than in negative patients (50.8%vs 17.6%, P = 0.03), and were the independent factors for predicting SIBO in postgastrectomy patients (P = 0.02). The prevalences of dumping syndrome and hypoglycemia after oral glucose were 35 (46.1%) and 19 (25.0%), and were not different between both groups. However, the plasma glucose was significantly lower in SIBO-positive than in SIBO-negative patients at 120 and 150 min after oral glucose load (P < 0.05). No significant differences were observed in pulse rate and hematocrit in both groups. CONCLUSIONS & INFERENCES: SIBO is common among postgastrectomy patients. It appears to be associated with postprandial intestinal symptoms and might aggravate late hypoglycemia. SIBO could be a new therapeutic target for managing intestinal symptoms in postgastrectomy patients.

Novel hMLH1 and hMSH2 germline mutations in African Americans with colorectal cancer.

CONTEXT: Germline mutations of the DNA mismatch repair (MMR) genes hMLH1 and hMSH2 have been shown to cosegregate with the colorectal cancer phenotype in multiple hereditary nonpolyposis colorectal cancer (HNPCC) pedigrees. However, the frequency of these mutations among African American patients with colorectal cancer is unknown. OBJECTIVE: To investigate the frequency of germline alterations of the DNA MMR genes hMLH1 and hMSH2 among African Americans affected by HNPCC and early-age onset colorectal cancer. DESIGN, SETTING, AND PATIENTS: Forty unrelated African American HNPCC and early-age onset colorectal cancer patients (8 women, 3 men) were identified from the cancer registry at a National Cancer Institute-designated referral center, 11 of whom were available for and agreed to study participation from January 1997 to February 1998. The mean age of the subjects was 44 years. An additional 50 age- and sex-matched African Americans without personal or family history of colorectal, endometrial, ovarian, urinary tract, or upper gastrointestinal tract malignancy were also studied as a polymorphism control population. In all subjects, genomic DNA was amplified by polymerase chain reaction for all hMLH1 and hMSH2 exons and screened using single-strand conformation polymorphism (SSCP) analysis. Samples demonstrating significant SSCP shifts underwent automated nucleotide sequencing analysis. MAIN OUTCOME MEASURE: Frequency of hMLH1 and hMSH2 germline alterations in the affected and control subjects. RESULTS: Germline hMLH1 and hMSH2 mutations were detected in 3 (27%) of the African American colorectal cancer probands studied. Each mutation was novel. Two hMLH1 (an A-->T transversion at codon 26 and a GG-->AT substitution across codons 177 and 178) mutations and 1 hMSH2 mutation (a C-->T transition at codon 389) were identified in 3 female study subjects. Six other hMLH1 and hMSH2 alterations were detected but were presumed to be polymorphisms. Neither missense mutation (at codons 26 and 389) was detected in the control population. CONCLUSIONS: The results of our analysis support an association between the 3 mutations reported and predisposition to colorectal cancer. Further studies are needed to define DNA MMR gene-associated colorectal cancer in African Americans, an understudied population at increased risk of fatal colorectal cancer.

Effect of weight loss on resting energy expenditure in hypertensive and normotensive obese women.

Hyperinsulinemia and the associated increased sympathetic nervous activity have been proposed to implicate in the development of hypertension and obesity. The role of hyperinsulinemia in mediated resting energy expenditure (REE) in hypertensive obese subjects is not clear. The effect of weight loss on REE between hypertensive and normotensive obese women are also unknown. We measured fasting plasma glucose, insulin, lipids concentrations, REE and body composition by bioelectrical impedance methods before and after a weight loss program in 9 newly diagnosed hypertensive and 10 normotensive obese women. As compared with age-matched lean control women (n=14), obese subjects had higher fasting plasma glucose, insulin concentrations and REE values. However, these variables were not different between obese groups. Although REE and fasting plasma insulin concentrations correlated well in simple correlation (r=0.708, p<0.001), this relationship disappeared after adjusting for values of fat free mass (FFM). Weight loss for approximately 10% of initial weight led to significant decreases of blood pressure and fasting plasma insulin concentrations in both obese groups. Fasting plasma cholesterol, LDL cholesterol and triglyceride concentrations decreased in hypertensive obese individuals. Significant fall of REE in hypertensive group (p<0.05) and normotensive group (p<0.02) were observed following weight loss. However, the ratio of REE to FFM decreased significantly only in hypertensive subjects (114.6+/-5.2 KJ/day x Kg(-1) to 107.2+/-4.6 KJ/day x Kg(-1), p<0.05). In conclusion, obese women, either hypertensive or normotensive, had higher fasting plasma insulin concentrations and REE than those of lean controls, although these variables were not different between obese groups. No significant relation between fasting plasma insulin and REE could be found. Weight loss produced a significant decrease of REE/FFM only in hypertensive obese women. Further study to evaluate the effect of weight loss on energy expenditure in hypertensive obese subjects is necessary.

Effect of weight loss on blood pressure and insulin resistance in normotensive and hypertensive obese individuals.

In this study, the effect of weight loss on blood pressure and various facets of glucose and insulin metabolism was examined in 22 subjects with mild to moderate obesity; 11 with high blood pressure (diastolic blood pressure > 95 mm Hg) and 11 with normal blood pressure (diastolic blood pressure < 90 mm Hg). The two groups were similar in mean (+/- SEM) body mass index at baseline (30.2 +/- 1.0 v 31.6 +/- 1.1 kg/m2), and each group lost approximately 8 kg during the 3-month study period. Blood pressure fell significantly (P < .003) following the 8 kg weight loss in both the normotensive (122 +/- 3/81 +/- 3 to 110 +/- 3/74 +/- 2 mm Hg) and hypertensive (149 +/- 3/98 +/- 1 to 135 +/- 3/86 mm Hg) subjects. Furthermore, the plasma glucose and insulin responses to a 75 g oral glucose load were significantly lower (P < .001) following weight loss. Finally, insulin resistance, as assessed by determining the steady-state plasma glucose (SSPG) concentration at the end of a 180 min infusion of somatostatin, insulin, and glucose, was also lower (P < .002) after the 8 kg weight loss in the normotensive (243 +/- 23 to 172 +/- 15 mg/dL) and hypertensive subjects (266 +/- 18 to 181 +/- 25 mg/dL). Since the steady-state plasma insulin concentrations were, if anything, slightly lower after weight loss in both groups, the lower post-weight loss SSPG values actually underestimate the improvement of insulin resistance. Thus, weight loss of 8 kg in moderately obese individuals leads to significant decreases in blood pressure and plasma glucose and insulin concentrations in response to an oral glucose challenge and degree of insulin resistance.

A brain L-type calcium channel alpha 1 subunit gene (CCHL1A2) maps to mouse chromosome 14 and human chromosome 3.

A rat brain cDNA probe was used to localize a gene encoding the alpha 1 subunit of neuronal dihydropyridine-sensitive L-type calcium channels in the mouse and human genomes. Hybridization of the probe to Southern blots made with DNAs from a Chinese hamster x mouse somatic cell hybrid panel indicated that this gene maps to mouse chromosome 14 (Chr 14). Southern blot analysis of an intersubspecies cross demonstrated that the calcium channel alpha 1 subunit gene, termed Cchl1a2, can be positioned 7.5 cM proximal to Np-1. Similarly, segregation among human X rodent somatic cell hybrids indicated that CCHL1A2 maps to human chromosome 3. These assignments are consistent with a region of linkage homology between human chromosome 3p and a proximal region of mouse Chr 14.

Biogenesis of transverse tubules in skeletal muscle in vitro.

The transverse (T) tubules of skeletal muscle are membrane tubules that are continuous with the plasma membrane and penetrate the mature muscle fiber radially to carry surface membrane depolarization to the sites of excitation-contraction coupling. We have studied the development of the T-tubule system in cultured amphibian and mammalian muscle cells using a fluorescent lipid probe and antibodies against T-tubules and plasma membranes. Both the lipid probe and the T-tubule antibody recognized an extensive tubular membrane system which subsequently differentiated into the T-system. At all developmental stages, the molecular composition of the T-system was distinct from that of the plasma membrane, suggesting that during myogenesis T-tubules and the plasma membrane form independently from each other and that exchange of membrane proteins between the two continuous compartments is restricted. In rat muscle cultures, T-tubule-specific antigens were first expressed in terminally differentiated myoblasts. Prior to myoblast fusion the antigens appeared as punctate label throughout the cytoplasm. Shortly after fusion the T-tubule-specific antibody labeled a tubular membrane system that extended from the perinuclear region and penetrated most parts of the cells. In contrast, the lipid probe, which labels the T-tubules by virtue of their direct continuity with the plasma membrane, only labeled short tubules extending from the plasma membrane into the periphery of the myotubes at the early stage in development. Thus, the assembly of the T-tubules appears to begin before their connections with the plasma membrane are established.

A membrane form of guanylate cyclase is an atrial natriuretic peptide receptor.

Atrial natriuretic peptide (ANP) is a polypeptide hormone whose effects include the induction of diuresis, natriuresis and vasorelaxation. One of the earliest events following binding of ANP to receptors on target cells is an increase in cyclic GMP concentration, indicating that this nucleotide might act as a mediator of the physiological effects of the hormone. Guanylate cyclase exists in at least two different molecular forms: a soluble haem-containing enzyme consisting of two subunits and a non-haem-containing transmembrane protein having a single subunit. It is the membrane form of guanylate cyclase that is activated following binding of ANP to target cells. We report here the isolation, sequence and expression of a complementary DNA clone encoding the membrane form of guanylate cyclase from rat brain. Transfection of this cDNA into cultured mammalian cells results in expression of guanylate cyclase activity and ANP-binding activity. The ANP receptor/guanylate cyclase represents a new class of mammalian cell-surface receptors which contain an extracellular ligand-binding domain and an intracellular guanylate cyclase catalytic domain.

Purification of murine erythropoietin produced in serum-free cultures of erythroleukemia cells.

We previously documented that several erythroleukemia cell lines released factors that stimulated erythropoiesis in vivo and in vitro. A simple five-step scheme has been devised that allows purification of this erythropoietic activity to apparent homogeneity. The methods employed included lectin affinity chromatography (wheat germ agglutinin), gel filtration (ultro gel ACA44), ion exchange, hydroxylapatite, and high performance liquid chromatography. Following polyacrylamide gel electrophoresis, biologic activity was recovered in an area corresponding to a molecular weight of 35,000 daltons. Silver staining of a polyacrylamide gel after electrophoresis of our most purified preparation revealed a single band at 35,000 daltons.

Modulation of Chloroplast Fructose-1,6-bisphosphatase Activity by Light.

Inhibitor experiments indicate that light effect mediator(II) which is reductively activated by transfer of electrons from the photosynthetic electron transport system at or beyond ferredoxin, is involved in activation by light of fructose-1,6-bisphosphatase in the pea plant. Activation proceeds optimally when the pH is low and Mg(2+) is 10 millimolar. Modulation by light results in increases in maximal velocity, apparently as a result of changes in enzyme conformation. Pea leaf thylakoids are effective in modulating the activity of glyceraldehyde-3-phosphate dehydrogenase but not of fructose-1,6-bisphosphatase or glucose-6-phosphate dehydrogenase in Kalanchoë stromal extracts. There is apparently species specificity for modulation of some, but not all, of the modulatable enzymes.