Chronic unloading by left ventricular assist device reverses contractile dysfunction and alters gene expression in end-stage heart failure.

BACKGROUND: Left ventricular (LV) assist devices (LVADs) can improve contractile strength and normalize characteristics of the Ca(2+) transient in myocytes isolated from failing human hearts. The purpose of the present study was to determine whether LVAD support also improves contractile strength at different frequencies of contraction (the force-frequency relationship [FFR]) of intact myocardium and alters the expression of genes encoding for proteins involved in Ca(2+) handling. METHODS AND RESULTS: The isometric FFRs of LV trabeculae isolated from 15 patients with end-stage heart failure were compared with those of 7 LVAD-supported patients and demonstrated improved contractile force at 1-Hz stimulation, with reversal of a negative FFR after LVAD implantation. In 20 failing hearts, Northern blot analysis for sarcoplasmic endoreticular Ca(2+)-ATPase subtype 2a (SERCA2a), the ryanodine receptor, and the sarcolemmal Na(+)-Ca(2+) exchanger was performed on LV tissue obtained before and after LVAD implantation. These paired data demonstrated an upregulation of all 3 genes after LVAD support. In tissue obtained from subsets of these patients, Western blot analysis was performed, and oxalate-supported Ca(2+) uptake by isolated sarcoplasmic reticular membranes was determined. Despite higher mRNA for all genes after LVAD support, only SERCA2a protein was increased. Functional significance of increased SERCA2a was confirmed by augmented Ca(2+) uptake by sarcoplasmic reticular membranes isolated from LVAD-supported hearts. CONCLUSIONS: LVAD support can improve contractile strength of intact myocardium and reverse the negative FFR associated with end-stage heart failure. The expression of genes encoding for proteins involved in Ca(2+) cycling is upregulated (reverse molecular remodeling), but only the protein content of SERCA2a is increased.

Vaccination of young lambs against infection with Nematodirus battus using gamma irradiated larvae.

Helminthologically naIve 6-week-old Suffolk lambs were given 1-3 doses of 20000 gamma-irradiated infective larvae (L3) of the nematode Nematodirus battus at weekly intervals. Following an anthelmintic drench they were challenged with 50000 viable L3 at 10 weeks of age. Nematode worm burdens 14 days post-challenge showed a significant (P<0.01) 66% reduction in the single vaccine dose group. The two and three dose groups had mean worm burdens which were 30 and 42% lower than controls, respectively, although these were not statistically significant. There was little measurable stimulation of the immune system in the vaccinated lambs, suggesting that the repeatedly dosed animals may have developed immunological unresponsiveness to the parasite.

Neonicotinoid insecticides: molecular features conferring selectivity for insect versus mammalian nicotinic receptors.

The favorable selective toxicity of neonicotinoid insecticides (represented here by imidacloprid, thiacloprid, and a nitromethylene analogue) for insects versus mammals is not shared by three of their N-unsubstituted imine derivatives or by nicotine or epibatidine. The same selectivity pattern is evident at the receptor level, i.e., the insect nicotinic acetylcholine receptor (nAChR) versus mammalian nAChR subtypes (alpha1, alpha3, alpha4, and alpha7) assayed independently. The insect-selective compounds are not protonated with a nitroimine, cyanoimine, or nitromethylene group and the mammalian-selective compounds are ionized at physiological pH. We propose that the negatively charged tip of the nitro or cyano group (not a partial positive charge at imidazolidine N-1 as suggested earlier) interacts with a putative cationic subsite of the insect nAChR. This contrasts with the mammalian nAChRs where the iminium cation (+C-NH2 <--> C =+NH2) of the neonicotinoid imine derivatives or ammonium nitrogen of nicotine or epibatidine interacts with the anionic subsite.

Neurologic findings in coronary artery bypass patients: perioperative or preexisting?

OBJECTIVES/DESIGN: This prospective study compares the incidence of preexisting neurologic findings in elective cardiac surgery patients presenting with and without coronary atherosclerosis. SETTING: This single-center study was conducted at a tertiary care hospital. PARTICIPANTS/INTERVENTIONS: After Review Board approval and obtaining written informed consent, 11 patients undergoing valvular heart surgery, 9 patients undergoing similar valvular procedures with concomitant coronary artery bypass surgery, and 4 patients undergoing coronary artery bypass surgery alone were enrolled. Preoperatively, all patients underwent a structured neurologic assessment, and the latter four additionally had preoperative magnetic resonance imaging. MEASUREMENTS AND MAIN RESULTS: The patients, 9 of 24 of whom were female, were aged 46 to 78 years and, other than ischemic heart disease, had medical histories that were similar between groups, with the exception of one patient having scleroderma. None of the patients had a clinical history of neurologic or cerebrovascular disease. Nine percent (1 of 11) of the valve-only patients showed subtle preoperative neurologic abnormalities, compared with 89% (eight of nine) of the valve patients having concomitant coronary surgery and 100% (four of four) of coronary artery bypass-only patients. Additionally, brain imaging scans of all four coronary bypass patients showed nonspecific changes reported as scattered punctate areas of high signal less than 3 to 4 mm in diameter. CONCLUSION: This survey shows that both subtle neurological abnormalities and magnetic resonance imaging lesions can be found in a high percentage of patients with coronary atherosclerosis. Furthermore, this study indicates that without a standardized preoperative neurological examination, postoperative neurologic dysfunction cannot necessarily be ascribed to perioperative events.

Vasoactive intestinal polypeptide-like and peptide histidine isoleucine-like proteins excreted/secreted by Nippostrongylus brasiliensis, Nematodirus battus and Ascaridia galli.

Vasoactive intestinal polypeptide (VIP)-like protein was detected by dot blot analysis in the excretions/secretions (E/S) of Nematodirus battus and Ascaridia galli and was confirmed in the E/S of Nippostrongylus brasiliensis. ELISA analysis showed that N. brasiliensis E/S contained the highest proportion of VIP-like protein (28.04 pmoles/mg of total E/S protein) and A. galli E/S contained the lowest (10.89 pmoles/mg of total E/S protein). Peptide histidine isoleucine (PHI)-like protein was detected by dot blot analysis in the E/S products of N. brasiliensis, N. battus and A. galli. ELISA analysis suggested that A. galli E/S contained the highest proportion of PHI (20.77 nmoles/mg of total E/S protein) and N. battus E/S contained the lowest (0.67 nmoles/mg of total E/S protein). The possible significance of VIP-like and PHI-like substances in the E/S of gastrointestinal nematodes is discussed.

Why do some nematode parasites of the alimentary tract secrete acetylcholinesterase?

Many gastrointestinal nematodes secrete large amounts of acetylcholinesterases. Antibodies are produced against these secreted acetylcholinesterases and appear to give some protection against infection with some nematodes. The theory that acetylcholinesterase secreted by gastrointestinal nematodes may act as a biochemical holdfast by reducing contractions of the alimentary system has not been substantiated; a vasoactive intestinal polypeptide-like protein is secreted by some species and may be the biochemical holdfast. Secreted acetylcholinesterases may alter host cell permeability, have an anti-coagulant role, affect glycogenesis, and/or be important in certain aspects of acetate and choline metabolism. Probably the most important role for acetylcholinesterase secreted by nematodes is immune modulation and/or reduction of inflammation in the vicinity of the nematode. The reason why some species of gastrointestinal nematodes resistant to benzimidazoles contain elevated amounts of acetylcholinesterase is unclear.

A vasoactive intestinal polypeptide-like protein excreted/secreted by Nippostrongylus brasiliensis and its effect on contraction of uninfected rat intestine.

The 50-30 kDa fraction isolated from the excretory/secretory products (E/S) of the nematode Nippostrongylus brasiliensis significantly decreased the amplitude of contraction of segments of uninfected rat intestine when injected into the lumen of the segments maintained in an organ bath. Dot blot analysis of the fraction suggested that it was similar in immunoreactivity to porcine vasoactive intestinal polypeptide (VIP). When antiserum to porcine VIP was mixed with N. brasiliensis E/S and the mixtures were injected into the lumen of segments of rat intestine, the inhibitory effect of the E/S on amplitude of contraction decreased. When physiological concentrations of porcine VIP (12.9 pmol/ml) were injected into the lumen of segments of uninfected rat intestine the amplitude of contraction decreased significantly. Western blot analysis of the E/S, using antiserum to porcine VIP, recognized a 30 kDa protein in the E/S and also in whole worm homogenate suggesting that synthesis of the peptide occurs inside the nematode. Peptide histidine isoleucine (PHI)-like immunoreactivity was detected in a 68 kDa fraction of the E/S and the homogenate but this fraction did not affect the amplitude of contractions of the intestine.

[Outcome and cost of mechanical ventilation in patients with inoperable solid tumors and hematologic malignancies].

The application of mechanical ventilation can be life-saving in patients with acute respiratory failure, but is frequently ineffective in terminally ill patients. To analyze the outcome and cost of mechanical ventilation for patients with malignancies, we retrospectively reviewed 115 patients with inoperable solid tumors or hematologic malignancies who received mechanical ventilation. Eighty eight of these 115 patients (77%) died while on mechanical ventilation; 18 patients (16%) were weaned from mechanical ventilation but still died during their hospitalization. Six patients (5%) were discharged but survived less than 3 months and only three patients (3%) survived for more than 3 months after discharge. The total time of mechanical ventilation was 772 person-days, which represented 5.5% (772/13974) of the total mechanical ventilator use in our hospital in 1993. The total hospital cost from initiation of mechanical ventilation to discharge or death of these 115 patients was NT $11,238,022 (US $432,200) and daily hospital cost per person during mechanical ventilation was approximately NT $10,845 (US $420). The "daily cost of discharge survival" (defined as total hospitalization cost from the initiation of mechanical ventilation to discharge or death divided by total discharge person-days) for patients with inoperable solid tumors was NT $73,421 (US $2,820)/per person-day. We concluded that mechanical ventilation should not be routinely used in patients with inoperable solid tumors and hematologic malignancies. Physicians should explain the poor outcome to these patients and their families in advance and inquire about their willingness to forego mechanical ventilation.

Renal calcification: a complication of dexamethasone therapy in preterm infants with bronchopulmonary dysplasia.

We have previously reported a case study in which renal calcification formation may have been a complication of dexamethasone exposure in an infant with bronchopulmonary dsyplasia. To determine whether dexamethasone is associated with renal calcification formation, we conducted a prospective, nonrandomized study of 36 infants < 30 weeks' gestation and weighing < 1250 gm treated with dexamethasone because of bronchopulmonary dysplasia and compared them with a group not receiving dexamethasone. We identified seven infants in the dexamethasone group (n = 19) but no infant in the comparison group (n = 17) with renal calcifications at 2 months of age (p = 0.008). The urinary calcium excretion tended to be increased (15.5 +/- 16.6 vs 6.9 +/- 6.7 mg/kg/day ¿p = 0.05¿) and the calcium/creatinine ratio was significantly greater in the dexamethasone group (1.2 +/- 1.0 vs 0.6 +/- 0.4 ¿p = 0.02¿). Infants who received dexamethasone were significantly smaller 819.1 +/- 141.1 vs 954.6 +/- 141 gm ¿p = 0.008¿), were younger (26.2 +/- 1.7 vs 27.7 +/- 1.2 weeks ¿p = 0.004¿), received ventilator support longer (33.3 +/- 14.7 vs 12.1 +/- 14.7 days ¿p = 0.0001¿), and required more days of supplemental oxygen (54.3 +/- 9.7 vs 36.4 +/- 23.8 days ¿p = 0.009¿). We conclude that smaller, younger, and sicker infants are at the highest risk for the development of renal calcifications and that dexamethasone may be associated with increased urinary calcium excretion, which contributes to renal calcification formation.

The presence of peptides related to the adipokinetic hormone/red pigment-concentrating hormone family in the nematode, Panagrellus redivivus.

Immunocytochemistry using polyclonal antisera raised to fragments or derivatives of locust adipokinetic hormone (AKH) I and IIs (Schooneveld et al., 1983, 1985, 1986) selectively stained cells in the nervous system of the free-living nematode, Panagrellus redivivus. Antiserum 528 (raised to the C-terminus of AKH IIs) stained the dorsal cephalic papillary cell bodies and the anterior nerve ring. Fibres in the lateral cords were stained with antiserum 241 that recognises the C-terminus of AKH I. Substances reacting to antisera 433 (raised to the N-terminal sequence of AKH I and IIs) 528 and 241 were present in the preanal ganglion and associated ventral nerve fibres. In males, all three antisera stained fibres leading to the base of the spicules. A peptide fraction from whole P. redivivus evoked an adipokinetic response in the locust, Schistocera gregaria which was dose dependent and was abolished by treatment with endopeptidase 24:11 but not by boiling or by incubation with leucine aminopeptidase. The adipokinetic activity was reduced by over 70% on incubation of the peptide fraction with pyroglutamyl aminopeptidase. The same fraction induced hyperglycaemia when injected into the cockroach, Periplaneta americana. These results are consistent with the existence in P. redivivus of peptides that are structurally related to the arthropod adipokinetic hormone/red pigment-concentrating hormone (AKH/RPCH) family.

Trichinella spiralis: antigenic epitopes from the stichocytes detected in the hypertrophic nuclei and cytoplasm of the parasitized muscle fibre (nurse cell) of the host.

Monoclonal antibodies raised against antigens present in the excretions/secretions (E/S) of larval Trichinella spiralis, polyclonal antibodies raised against E/S and antisera from rabbits and pigs infected with T. spiralis were used in conjunction with immunocytochemical techniques to detect antigens in sections of muscle from mice that had been infected with T. spiralis for 15 or 30 days. The antibodies recognized epitopes in the stichocytes, on the surface of the cuticle, in the lumen of the oesophagus and in the lumen of the intestine of encysted larvae. Monoclonal antibodies 7C2C5 and 1H7 and the polyclonal antibodies recognized epitopes in the cavity occupied by the larva, in the cytoplasm of the nurse cell, and in the hypertrophic nuclei of the nurse cell, but did not recognize material in the smaller nuclei of the nurse cell, in the cyst wall or in the surrounding muscle. Monoclonals 3B2E6 and 1D11G8B2, which recognized epitopes in the stichocytes and on the surface of the cuticle of the larvae, gave negative results with the cytoplasm and nuclei of the nurse cell. A polyclonal antibody raised against Trichuris suis recognized epitopes in the muscle and hypodermis of encysted T. spiralis but gave negative results with material in the nurse cell and nurse cell nuclei. The possibility that the antigen detected in the cytoplasm and nuclei of the nurse cell is produced by the stichocytes of the nematode and that it is controlling genes of the altered muscle fibre, either directly or indirectly, is discussed.

A copia-like element in Pisum demonstrates the uses of dispersed repeated sequences in genetic analysis.

A DNA sequence between two legumin genes in Pisum is a member of the copia-like class of retrotransposons and represents one member of a polymorphic and heterogeneous dispersed repeated sequence family in Pisum. This sequence can be exploited in genetic studies either by RFLP analysis where several markers can be scored together, or the segregation of individual elements can be followed after PCR amplification of specific members.

Monooctanoin dissolution of gallstones via a cholecystostomy tube.

Monooctanoin was infused through a cholecystostomy tube to dissolve multiple gallstones and re-establish cystic duct patency in a patient who was not a candidate for surgery.

Changes in the adenylate energy charge of Nippostrongylus brasiliensis and Nematodirus battus during the development of immunity to these nematodes in their host.

Infection of rats with 2000 infective juveniles of Nippostrongylus brasiliensis and of lambs with 60 000 infective juveniles of Nematodirus battus results in a well-marked immunity to these nematodes in their respective host. There is a fall in the adenylate energy charge value of these nematodes during the course of these infections, reaching values of 0.37 in males and 0.27 in females of N. brasiliensis, and 0.31 in males and 0.23 in females of N. battus towards the end of the infections. In hosts given relatively small numbers of infective juveniles, the values for the nematodes removed from the hosts late in the infection remain at a relatively high level. These results indicate that the immune response of the host may affect the energy status of these nematodes, and this could help to explain their subsequent expulsion from the immune host.

Interactions in vitro between sporozoites of Eimeria tenella and host peritoneal exudate cells: electron microscopal observations.

The interactions of sporozoites of Eimeria tenella with peritoneal cells from normal and from immunized chickens were examined in vitro. Although the uptake of sporozoites by cells from immunized birds was greater than by cells from susceptible birds, no differences were apparent in their appearance in electron micrographs. Entry into both macrophages and heterophils (comparable to mammalian neutrophils) was by phagocytosis. The findings are discussed.