Impaired cardiac performance in heterozygous mice with a null mutation in the sarco(endo)plasmic reticulum Ca2+-ATPase isoform 2 (SERCA2) gene

The sarco(endo)plasmic reticulum Ca2+-ATPase isoform 2 (SERCA2) gene encodes both SERCA2a, the cardiac sarcoplasmic reticulum Ca2+ pump, and SERCA2b, which is expressed in all tissues. To gain a better understanding of the physiological functions of SERCA2, we used gene targeting to develop a mouse in which the promoter and 5' end of the gene were eliminated. Mating of heterozygous mutant mice yielded wild-type and heterozygous offspring; homozygous mutants were not observed. RNase protection, Western blotting, and biochemical analysis of heart samples showed that SERCA2 mRNA was reduced by approximately 45% in heterozygous mutant hearts and that SERCA2 protein and maximal velocity of Ca2+ uptake into the sarcoplasmic reticulum were reduced by approximately 35%. Measurements of cardiovascular performance via transducers in the left ventricle and right femoral artery of the anesthetized mouse revealed reductions in mean arterial pressure, systolic ventricular pressure, and the absolute values of both positive and negative dP/dt in heterozygous mutants. These results demonstrate that two functional copies of the SERCA2 gene are required to maintain normal levels of SERCA2 mRNA, protein, and Ca2+ sequestering activity, and that the deficit in Ca2+ sequestering activity due to the loss of one copy of the SERCA2 gene impairs cardiac contractility and relaxation.     

Determination of erythromycin in urine and plasma using microbore liquid chromatography with tris(2,2''-bipyridyl)ruthenium(II) electrogenerated chemiluminescence detection

Neuroendocrine lung tumors have been considered by some to be a continuum ranging from relatively benign typical carcinoids to highly malignant small-cell carcinomas. Histopathological diagnosis may sometimes be difficult because of their overlapping features. Correct classification, however, carries important prognostic and therapeutic significance. To determine the clinicopathological implications of retinoblastoma (RB) protein expression in these neoplasms, we examined the RB status in a series of neuroendocrine tumors by immunohistochemical analysis of paraffin-embedded tissue sections. A total of 105 tumors were studied. All 44 typical and 15 atypical carcinoids, one of which was initially misdiagnosed as a small-cell carcinoma, manifested a heterogeneous RB-positive staining pattern. Atypical carcinoids in general showed an increase in the number of tumor cells with strong nuclear staining compared to typical carcinoids. In contrast, all 40 small-cell and 6 large-cell neuroendocrine carcinomas failed to show RB staining in any tumor nuclei, indicating loss of RB function. Our results suggest that RB status as measured by immunohistochemical staining can be used as a marker to distinguish typical and atypical carcinoids from small-cell and large-cell neuroendocrine carcinomas.     

Influence of mode and carbohydrate on the cytokine response to heavy exertion

1. This experiment was carried out to evaluate the productive and physiological consequences of a slight but long term food restriction of male broiler chickens from 2 commercial strains. 2. Cobb-500 and Ross chickens were submitted to a 20% food restriction from 8 to 21 d of age. Strain, food programme and their interactive effects were analysed in terms of consequences upon performance, mortality, incidence of sudden death syndrome (SDS) and ascites syndrome (AS), index of right cardiac hypertrophy and plasma concentrations of hormones related to metabolism and growth (T3, T4, T3:T4 ratio, IGF-I and GH). 3. Although some catch-up growth was observed by refeeding previously restricted birds after 22 d of rearing, food restriction decreased (P < or = 0.05) body weight at market age (42 d) irrespective of the strain, but improved (P < or = 0.05) food conversion. 4. The incidence of mortality was not high in non-restricted birds but SDS and AS caused more than 50% of deaths. Hypertrophic cardiac index was observed in chickens of both strains after 4 weeks of age and was higher in ad libitum fed birds. 5. During the period of food restriction, plasma T3 and IGF-I concentrations decreased whereas plasma T4 and GH concentrations increased compared to those of the age-matched ad libitum fed counterparts. During the subsequent ad libitum feeding period, few differences in circulating hormone concentrations were observed, except for the higher mean GH litres in previously food-restricted chickens at 35 d of age. 6. These results indicate that even a non-severe food restriction negatively affects body weight of 42-d-old male broilers but these are benefits with improved food efficiency and diminished mortality from metabolic disturbances. The hormone results suggest that the degree of food restriction applied was not severe because there was a very fast adaptive response with small and transient alterations in T3, T4 and GH plasma concentrations during the period of compensatory growth.     

Cadherins promote skeletal muscle differentiation in three-dimensional cultures

The cell-cell adhesion molecule N-cadherin, with its associated catenins, is expressed by differentiating skeletal muscle and its precursors. Although N-cadherin's role in later events of skeletal myogenesis such as adhesion during myoblast fusion is well established, less is known about its role in earlier events such as commitment and differentiation. Using an in vitro model system, we have determined that N-cadherin- mediated adhesion enhances skeletal muscle differentiation in three-dimensional cell aggregates. We transfected the cadherin-negative BHK fibroblastlike cell line with N-cadherin. Expression of exogenous N-cadherin upregulated endogenous beta-catenin and induced strong cell-cell adhesion. When BHK cells were cultured as three-dimensional aggregates, N-cadherin enhanced withdrawal from the cell cycle and stimulated differentiation into skeletal muscle as measured by increased expression of sarcomeric myosin and the 12/101 antigen. In contrast, N-cadherin did not stimulate differentiation of BHK cells in monolayer cultures. The effect of N-cadherin was not unique since E-cadherin also increased the level of sarcomeric myosin in BHK aggregates. However, a nonfunctional mutant N-cadherin that increased the level of beta-catenin failed to promote skeletal muscle differentiation suggesting an adhesion-competent cadherin is required. Our results suggest that cadherin-mediated cell-cell interactions during embryogenesis can dramatically influence skeletal myogenesis.     

Phenotype resembling Gitelman's syndrome in mice lacking the apical Na+-Cl- cotransporter of the distal convoluted tubule

Mutations in the gene encoding the thiazide-sensitive Na+-Cl- cotransporter (NCC) of the distal convoluted tubule cause Gitelman's syndrome, an inherited hypokalemic alkalosis with hypomagnesemia and hypocalciuria. These metabolic abnormalities are secondary to the deficit in NaCl reabsorption, but the underlying mechanisms are unclear. To gain a better understanding of the role of NCC in sodium and fluid volume homeostasis and in the pathogenesis of Gitelman's syndrome, we used gene targeting to prepare an NCC-deficient mouse. Null mutant (Ncc-/-) mice appear healthy and are normal with respect to acid-base balance, plasma electrolyte concentrations, serum aldosterone levels, and blood pressure. Ncc-/- mice retain Na+ as well as wild-type mice when fed a Na+-depleted diet; however, after 2 weeks of Na+ depletion the mean arterial blood pressure of Ncc-/- mice was significantly lower than that of wild-type mice. In addition, Ncc-/- mice exhibited increased renin mRNA levels in kidney, hypomagnesemia and hypocalciuria, and morphological changes in the distal convoluted tubule. These data indicate that the loss of NCC activity in the mouse causes only subtle perturbations of sodium and fluid volume homeostasis, but renal handling of Mg2+ and Ca2+ are altered, as observed in Gitelman's syndrome.     

Test characteristics of acridine orange, Gram, and May-Grünwald-Giemsa stains for enumeration of intracellular organisms in bronchoalveolar lavage fluid

For enumeration of intracellular organisms (ICO) in bronchoalveolar lavage fluid samples, the May-Grünwald-Giemsa (MGG) stain displayed higher interobserver agreement than the acridine orange and Gram stains. The MGG stain offered a reliable enumeration of ICO when 200 cells were counted by one observer.     

Exogenous surfactant and positive end-expiratory pressure in the treatment of endotoxin-induced lung injury

OBJECTIVE: To evaluate the efficacy of treating endotoxin-induced lung injury with single dose exogenous surfactant and positive end-expiratory pressure (PEEP). DESIGN: Prospective trial. SETTING: Laboratory at a university medical center. SUBJECTS: Nineteen certified healthy pigs, weighing 15 to 20 kg. INTERVENTIONS: Pigs were anesthetized and surgically prepared for hemodynamic and lung function measurements. Animals were randomized into four groups: a) Control pigs (n = 4) received an intravenous infusion of saline without Escherichia colilipopolysaccharide (LPS); b) the LPS group (n = 5) received an intravenous infusion of saline containing LPS (100 microg/kg); c) the PEEP plus saline group (n = 5) received an intravenous infusion of saline containing LPS. Two hours after LPS infusion, saline was instilled into the lung as a control for surfactant instillation, and the animals were placed on 7.5 cm H2O of PEEP; d) the PEEP plus surfactant group (n = 5) received an intravenous infusion of saline containing LPS. Two hours following LPS infusion, surfactant (50 mg/kg) was instilled into the lung and the animals were placed on 7.5 cm H2O of PEEP. PEEP was applied first and surfactant or saline was instilled into the lung while maintaining positive pressure ventilation. All groups were studied for 6 hrs after the start of LPS injection. At necropsy, bronchoalveolar lavage was performed and the right middle lung lobe was fixed for histologic analysis. MEASUREMENTS AND MAIN RESULTS: Compared with LPS without treatment, PEEP plus surfactant significantly increased PaO2 (PEEP plus surfactant = 156.6 +/- 18.6 [SEM] torr [20.8 +/- 2.5 kPa]; LPS = 79.2 +/- 21.9 torr [10.5 +/- 2.9 kPa]; p<.05), and decreased venous admixture (PEEP plus surfactant = 12.5 +/- 2.0%; LPS = 46.9 +/- 14.2%; p< .05) 5 hrs after LPS infusion. These changes were not significant 6 hrs after LPS infusion. PEEP plus surfactant did not alter ventilatory efficiency index (VEI = 3800/[peak airway pressure - PEEP] x respiratory rate x PacO2), or static compliance as compared with LPS without treatment at any time point. Cytologic analysis of bronchoalveolar lavage fluid showed that surfactant treatment significantly increased the percentage of alveolar neutrophils as compared with LPS without treatment (PEEP plus surfactant = 39.1 +/- 5.5%; LPS = 17.4 +/- 6.6%; p< .05). Histologic analysis showed that LPS caused edema accumulation around the airways and pulmonary vessels, and a significant increase in the number of sequestered leukocytes (LPS group = 3.4 +/- 0.2 cells/6400 micro2; control group = 1.3 +/- 0.1 cells/6400 micro2; p < .05). PEEP plus saline and PEEP plus surfactant significantly increased the total number of sequestered leukocytes in the pulmonary parenchyma (PEEP plus surfactant = 8.2 +/- 0.7 cells/6400 micro2; PEEP plus saline = 3.9 +/- 0.2 cells/6400 micro2; p <.05) compared with the control and LPS groups. CONCLUSIONS: We conclude that PEEP plus surfactant treatment of endotoxin-induced lung injury transiently improves oxygenation, but is unable to maintain this salutary effect indefinitely. Thus, repeat bolus dosing of surfactant or bolus treatment followed by continuous aerosol delivery may be necessary for a continuous beneficial effect.     

EXPERIMENTAL EVALUATION OF OPTIMAL MULTIVARIABLE SERVO CONTROL IN CONJUNCTION WITH CONVENTIONAL REGULATORY CONTROL†

The problem considered is that of driving a multivariable process controlled by conventional feedback control algorithms, from some initial state, to a specified final state such that a linear performance index is minimized. The formulation is based on a linear, discrete, time-invariant model of the process plus a minimum-time or sum of the absolute errors criterion and is solved using a standard linear programming algorithm. Constraints on the state and/or control variables can be incorporated to guarantee practical, physically realizable control policies. Experimental and simulated results show that the technique is easy to implement, and that the use of different criteria produces significantly different process transients. Experimental data from a computer-controlled, pilot plant evaporator show that the method is practical and produces better results than conventional methods.     

Diagnostic validity of the Eppendorf Schizophrenia Inventory (ESI): A self-report screen for ultrahigh risk and acute psychosis": Correction to Niessen et al. (2010).

Reports an error in "Diagnostic validity of the Eppendorf Schizophrenia Inventory (ESI): A self-report screen for ultrahigh risk and acute psychosis" by Maurice A. J. Niessen, Peter M. A. J. Dingemans, Reinaud van de Fliert, Hiske E. Becker, Dorien H. Nieman and Don Linszen (Psychological Assessment, 2010[Dec], Vol 22[4], 935-944). In the first full paragraph, the references to the numbers in Table 5 are incorrect in the sentence that begins "We then proceeded with calculating accuracy measures...". A corrected version of the sentence is presented in the erratum. Also presented in the erratum are corrections to variables mentioned elsewhere in the article. The aforementioned changes are very small and do not in any way affect the findings of the research. (The following abstract of the original article appeared in record 2010-24850-009.) Providers of mental health services need tools to screen for acute psychosis and ultrahigh risk (UHR) for transition to psychosis in help-seeking individuals. In this study, the Eppendorf Schizophrenia Inventory (ESI) was examined as a screening tool and for its ability to correctly predict diagnostic group membership (e.g., help seeking, mild psychiatric complaints, highly symptomatic mood or anxiety disorder, UHR, acute psychosis). Diagnostic evaluation with established instruments was used for diagnosis in 3 research samples. UHR status was assessed with the Structured Interview for Prodromal Symptoms/Scale of Prodromal Symptoms (Miller et al., 1999) and the Bonn Scale for the Assessment of Basic Symptoms Prediction list (Gross, Huber, Klosterk?tter, & Linz, 1987; Klosterk?tter, Hellmich, Steinmeyer, & Schulze-Lutter, 2001). This study showed that members of different diagnostic groups rate themselves significantly differently on the ESI and its subscales. A new subscale was constructed, the UHR–Psychosis scale, that showed good utility in detecting individuals with interview-diagnosed UHR status and acute psychosis. The scale is also sensitive to the threshold between UHR and acute psychosis. Practical applications of the ESI include use as a diagnostic tool within various settings. (PsycINFO Database Record (c) 2011 APA, all rights reserved)     

Carbohydrate supplementation and the lymphocyte proliferative response to long endurance running

This randomized, double-blind, placebo-controlled study examined the influence of 6% carbohydrate ingestion on hormonal and lymphocyte proliferative responses (5 total samples over 9 hours) to 2.5 h of high-intensity running by 30 experienced marathon runners. The T-cell response differed between groups, with the placebo group exhibiting a greater increase immediately post-run and greater decrease at 3 h of recovery. No group differences were observed for Con A-, PHA-, or PWM-induced lymphocyte proliferation. However, when PHA was adjusted per T-cell, group differences were observed, highlighted by a decrease in the placebo group immediately post-run. Glucose and cortisol responses differed between groups, with glucose lower and cortisol higher in the placebo group immediately post-run. Post-run glucose correlated negatively with postrun cortisol (r=-0.670, P< 0.001) and epinephrine (r=-0.540, P=0.002). Post-run cortisol also correlated negatively with total lymphocytes and T-cells at 1.5 hours (r=-0.429, P=0.018 and r=-0.424, P=0.019, respectively) and 3 hours (r=-0.566, P=0.001 and r=-0.523, P=0.003, respectively) of recovery. The pre- to post-run change in glucose correlated to the same changes in PHA/T-cell (r=0.456, P=0.011). The data support an interactive effect of carbohydrate ingestion on plasma glucose and cortisol. The data support an interactive effect of carbohydrate ingestion on plasma glucose and cortisol, T-cell trafficking, and cell-adjusted PHA-induced lymphocyte proliferation following long endurance running.