Endocardial endothelium selectively modifies relaxation in rat papillary muscle

The selective removal of endocardial endothelium of rat left ventricular papillary muscles by 1-second immersion in 0.5% Triton X-100 showed little influence on resting tension and only a small decrease in peak isometric tension (8.3 +/- 1.4 vs 9.6 +/- 2.4 mN/mm2 at Lmax, p > 0.05) with no reduction in maximal rate of tension development (+dT/dtmax; 136 +/- 21 vs 137 +/- 18 mN/mm2/s, p > 0.05). In contrast, there was a marked increase in maximal rate of tension decline (-dT/dtmax) from 71 +/- 14 to 92 +/- 15 mN/mm2/s (p < 0.05), so that the ratio between +dT/dtmax and -dT/dtmax fell from 1.98 +/- 0.27 to 1.51 +/- 0.13 (p < 0.01). Removal of endocardial endothelium led to a significant shortening of isometric twitch contractions. Time to peak tension was abbreviated from 111 +/- 20 to 84 +/- 8 ms (p < 0.05) and the half relaxation time from 92 +/- 9 to 68 +/- 8 ms (p < 0.01). Time to +dT/dtmax was also shortened from 31 +/- 6 to 44 +/- 9 ms (p < 0.05) and time to -dT/dtmax from 90 +/- 12 to 62 +/- 10 ms (p < 0.01). These effects were not influenced by alterations in stimulation frequency or muscle length. The early onset of relaxation and abbreviated duration of relaxation together with an increased rate of decline in tension led to a shorter total twitch which may explain the slightly lower peak tension once the endocardial endothelium was removed. Our findings confirm that endocardial endothelium modulates myocardial contraction, with a predominant influence on relaxation.     

Porcine pyridoxal kinase c-DNA cloning, expression and confirmation of its primary sequence

Porcine brain pyridoxal kinase has been cloned. A 1.2 kilo-based cDNA with a 966-base pair open reading frame was determined from a porcine brain cortex cDNA library using PCR technique. The DNA sequence was shown to encode a protein of 322 amino acid residues with a molecular mass of 35.4 kDa. The amino acid sequence deduced from the nucleotide sequence of the cDNA was shown to match the partial primary sequence of pyridoxal kinase. Expression of the cloned cDNA in E. coli has produced a protein which displays both pyridoxal kinase activity and immunoreactivity with monoclonal antibodies raised against natural enzyme from porcine brain. With respect to the physical properties, it is shown that the recombinant protein exhibits identical kinetic parameters with the pure enzyme from porcine brain. Although the primary sequence of porcine pyridoxal kinase has been shown to share 87% homology with the human enzyme, we have shown that the porcine enzyme carries an extra peptide of ten amino acid residues at the N-terminal domain.     

Hematological investigations in conditions of long-term space flights

In the nearly 15-month mission aboard MIR the cosmonaut-physician and members of three crews (MIR-15, -16, and -17) carried out a program of hematological investigations. Most of the changes related to the red blood system and included reduction in hemoglobin and hematocrit. Erythrocytes had decreased concentration and took on abnormal forms. There were also signs of altered metabolism of erythrocytes. Of interest are phase-by-phase variations in the levels of erythrocytes in the course of long-term stay in microgravity, and absence of a convincing correlation between hemoglobin, erythrocyte and hematocrit levels. But for lymphocytosis that returned to the norm already on the first day of recovery, no material changes occurred to the leukocyte profile. Investigations at the landing site displayed erythropenia, decreased reticulocytes and ensuing reticulocyte reaction, and gradual regain of the erythrocyte number that can be viewed as a normal physiological reaction of the blood system to the set of factors of spaceflight and early readaptation. Besides, the investigations showed a large individually of blood reactions to prolonged stay in space flight.     

Vascular injury during lumbar disc surgery. Report of two cases; a review of the literature

Iatrogenic vascular injuries are unusual complications of lumbar disc surgery. The incidence of such injuries is very low but probably underestimated because clinical manifestations may be extremely variable depending on the extension of trauma. Diagnosis is suspected when early signs of retroperitoneal haemorrhage appear, but may often be delayed for weeks or years due to formation of a pseudoaneurysm or an arteriovenous fistula which may be of gradual onset and produce initially only a few symptoms. Prompt diagnosis and aggressive treatment can improve the current mortality rate of more than 50%. Two cases are described that illustrate the full spectrum of acute and chronic manifestations of such injuries. One case of acute haemorrhage due to arterial trauma was immediately detected and the other case with arteriovenous fistula was recognized several years post-operatively.     

Heat shock gene expression during recovery after transient cold shock in Drosophila auraria (Diptera: Drosophilidae)

Newly synthesized polypeptides during recovery from prolonged cold treatment (0 degree to -1 degree C) of Drosophila auraria, a montium subgroup species, of the melanogaster species group, were analysed in denaturing polyacrylamide gels. In addition, during the cold shock recovery period, Northern analysis of the hsp83 mRNA was performed. A significant induction of two polypeptides, which exhibited electrophoretic mobilities, with the heat inducible 83 and 70 kD hsp83 and hsp70 was detected, but no such induction was evident in the so-called 'small' hsp genes. These results are compared and discussed with those observed in D. melanogaster.     

Effect of hemorrhagic shock on endotoxin-inducing TNF production and intra-tissue lipopolysaccharide-binding protein mRNA expression and their relationship

In order to further elucidate effect of hemorrhagic shock on endotoxin-inducing cytokine production, the present study was designed to investigate the production of tumor necrosis factor alpha (TNF alpha) induced by low-dose (1 microgram/kg) of lipopolysaccharide (LPS) and its cellular sources after hemorrhagic shock (HS) in rats. With combination of expression of lipopolysaccharide-binding protein (LBP) mRNA in the liver, lungs, and kidneys, we further analyzed a possible mechanism for increasing sensitivity to LPS by shock. We found in vivo that plasma TNF alpha levels in the HS + LPS group were 20-fold higher than those in the HS group (p < .01) and 2.7-fold higher than those in the LPS group (p < .05). It was shown in vitro that the capacity of the peripheral white blood cells to produce TNF alpha in response to LPS stimulation was significantly decreased by 126% (p < .01) and 57% (p < .05) compared with the pre-shock levels and sham group, respectively, at the end of resuscitation following shock, and still markedly inhibited 3 h after resuscitation, while the capacity of hepatic Kupffer's cells to produce TNF alpha was significantly increased by 110% compared with the sham group (p < .01) after shock and resuscitation. Results from RT-PCR showed that expression of LBP mRNA in the liver, lungs, and kidneys was increased after shock and resuscitation. It is suggested that hemorrhagic shock could significantly strengthen endotoxin to induce TNF alpha production, which might be due to up-regulation of LBP expression in tissues after shock, and the tissue macrophage population may be the main source for cytokine production in shock.     

Pharmacokinetics and retrograde colonic spread of budesonide enemas in patients with distal ulcerative colitis

METHODS: The retrograde spread of two budesonide enema formulations with different viscosities was investigated. The study design was open, randomized and two-period crossover. Three female and two male patients (age range: 35-45 years) with distal ulcerative colitis or proctitis participated. Both enema formulations contained a dose of 2 mg budesonide/100 mL. An unabsorbable radioactive marker (99mTc-labelled human serum albumin microcolloid) was added to the enema just before administration. All doses were given in the evening with the patients lying in a supine position during the whole investigation. The intestinal spread was followed for 8 h using scintigraphic imaging. Plasma samples for budesonide assay were taken during the 12 h after administration of the low viscosity enema. RESULTS: Budesonide plasma levels were measurable for up to 4-6 h. Cmax was 2.5 nmol/L (range: 0.9-4.5 nmol/L) and was attained in 1.5 h (range 1-3 h). The patients had no difficulty in retaining the enemas. There was a statistically significant difference in spread between the low and high viscosity enema. The low viscosity enema spread over an area situated between the rectum and the splenic flexure. The spread occurred mainly in the first 15 min after administration. In contrast, the high viscosity enema, in most cases, spread only over a minor part of this area and the rate and extent of spreading was also more variable with this formulation. CONCLUSION: The spread of a low viscosity enema appears to be well suited for the treatment of proctitis and distal colitis.