Effect of disuse on the ultrastructure of the achilles tendon in rats

We examined the influence exerted, through disuse of the hindlimb, on the collagen fibres of the achilles tendon in rats. With disuse the body mass decreased by 28%, and the mass of soleus muscle decreased by 20%. A decrease in the surface area and diameter was observed in the experimental group when compared to the control group. A histogram of the collagen fibres showed a decrease of the thick fibres in the experimental group. The maximum surface area of collagen fibres in the experimental group was seen to be only 43% of that of the control group. These results showed a decrease in the thickness of the collagen fibres of the achilles tendon through disuse. This seemed to suggest that resistance to tension is decreased by disuse.     

Evidence that thyroxine inhibits either basal or TRH-induced TSH secretion only after conversion to triiodothyronine

When TRH was administered every 15 min for 2 hr in euthyroid rats, equivalent modestly supraphysiologic doses of either T4 or T3 suppressed TRH-induced TSH secretion after 45 min. Pretreatment with iopanoic acid blocked the ability of T4 but not of T3 to suppress TRH-induced TSH secretion 2 hr after administration of the respective thyroid hormone. Pretreatment with iopanoic acid also blocked the ability of T4, but not of T3, to depress the elevated basal plasma TSH concentration of hypothyroid rats within 2 hr. Propylthiouracil did not significantly inhibit the ability of T4 to depress TRH-induced TSH secretion and only slightly depressed the ability of T4 to reduce the elevated plasma TSH of hypothyroid rats. Our data support the concept that although equivalent physiologic doses of T4 or T3 inhibit basal or TRH-induced TSH secretion equally rapidly, TSH inhibition produced by T4 is probably dependent on its rapid conversion to T3, either within the pituitary or peripherally. T3 thus seems to be exerting almost all the negative feedback effects on TSH secretion under the conditions of our experiments.     

Correlation between the virulence ofFrancisella tularensis in experimental mice and its acriflavine reaction

Correlation between the virulence of Francisella tularensis in experimental mice and its acriflavine reaction was studied. The cultures derived from all four strains (Ebina, CMB2, Schu, and N9) that had long been subcultured on agar media yielded two types of colonies, i.e., acriflavine reaction-positive (acf+) and acriflavine reaction-negative (acf-) colonies. All acf+ colonies, regardless of their parent strains, were shown to be low virulent in mice. Acf- colonies were shown to be either high (Ebina, CMB2) or low (Schu, N9) virulent. The low-virulent acf- colonies gained virulence during several passages in mice, whereas the acf+ colonies remained low virulent even after the animal passages.     

Effects of chemically defined tannins on poly (ADP-ribose) glycohydrolase activity.

Three classes of chemically defined tannins, gallotannins, ellagitannins and condensed tannins were examined for their inhibitory activities against purified poly (ADP-ribose) glycohydrolase. Ellagitannins showed higher inhibitory activities than gallotannins. In contrast, condensed tannins, which consist of an epicathechin gallate (ECG) oligomer without a glucose core were not appreciably inhibitory. Kinetic analysis revealed that the inhibition of ellagitannins was competitive with respect to the substrate poly(ADP-ribose), whereas gallotannins exhibited mixed-type inhibition. These results suggest that conjugation with glucose of hexahydroxy-diphenoyl (HHDP) group, which is a unique component of ellagitannins, potentiated the inhibitory activity, and that the structure of ellagitannins may have a functional domain which competes with poly(ADP-ribose) on the poly(ADP-ribose) glycohydrolase molecule.     

Two monoclonal antibodies against small-cell lung cancer show existence of synergism in binding

Summary Murine IgG1 monoclonal antibodies (mAbs), ITK-2 and ITK-3, were generated against a small-cell lung cancer (SCLC) cell line. Enzyme-linked immunosorbent assay using a variety of established cell lines as substrates, immunoperoxidase staining of freshly frozen tissue sections, and fluorescence-activated cell sorter analysis of peripheral blood leukocytes showed that these mAbs recognize a part of the SCLC-associated cluster 1 antigen. In immunoprecipitation studies, both ITK-2 and ITK-3 bound to a 145-kDa glycoprotein of SCLC cell membrane extracts, as did MOC-1 and NKH-1, which both recognize the cluster 1 antigen. However, because the binding of¹²⁵I-labeled ITK-2 to SCLC cells was not inhibited by MOC-1 or NKH-1, the binding site of ITK-2 on SCLC cells appeared to be different from that of either MOC-1 or NKH-1. Unexpectedly, binding of¹²⁵I-labeled ITK-2 to SCLC cells increased in the presence of ITK-3. This ITK-3-induced increase in ITK-2 binding was due partly to an increase in the number of binding sites for ITK-2 on SCLC cells. Addition of ITK-3 may, therefore, improve the effectiveness of ITK-2-based tumor detection or therapy.     

Mutants with Altered Ca2+-Channel Properties in PARAMECIUM TETRAURELIA: Isolation, Characterization and Genetic Analysis

Dancers are a group of mutants in Paramecium tetraurelia whose Ca²⁺ current inactivates poorly and are likely to be defective in the structure of their Ca²⁺ channels. These mutants show prolonged backward swimming in response to K⁺ and Ba ²⁺ in the medium and were selected by this property in a galvanotactic trough. The dancer mutants are semidominant, and all isolated mutants belong to one complementation group; they are not allelic to any of the previously isolated behavioral mutants of P. tetraurelia. The phenotypic change from the homozygous parent to heterozygous F₁ generation takes three to five fissions. There is no evidence of a cytoplasmic factor capable of converting the dancer to the wild-type phenotype, as has been demonstrated in the mutants pawn and cnr. We suggest that the dancer locus is a structural gene for the Ca²⁺ channel.     

Studies on the biological activity of an insulin-stimulating peptide from a tryptic digest of bovine serum albumin

Summary A two-chain polypeptide, which corresponds to amino acid residues 115–143 and 144–184(185) of bovine serum albumin, connected to each other by a disulfide bridge, potentiated the effects of insulin on glucose transport and glucose metabolism in isolated rat adipocytes. Although the peptide alone had little activity, it shifted the concentration-response curves of insulin-stimulated D-[I-¹⁴C]glucose oxidation, 2-deoxyglucose transport, and lipid synthesis from D-[U-¹⁴C]glucose to lower insulin concentrations. It also increased the maximal responses of these parameters to insulin. However, it did not affect insulin binding to adipocytes. The peptide protected insulin considerably from degradation, but this effect alone cannot account for its effect in increasing the maximal responses to the hormone, and even when degradation of a submaximal concentration of insulin was suppressed by bacitracin, the peptide still had an enhancing effect. These results suggest not only that the peptide influences a step distal to receptor-mediated insulin binding but also that inhibition of insulin degradation alone cannot explain its total effect.The peptide lost its insulin-stimulating activity completely when it was further digested with V8 or lysinespecific endopeptidase, or when it was reduced and then carboxamidomethylated or oxidized with performic acid. Similar active tryptic fragments were obtained from human and rat albumins.Insulin-stimulating peptides should be useful in studies on the mechanisms of insulin action including both the sensitivities and responsiveness of target cells to the hormone.Abbreviations ISP insulin-stimulating peptide - HEPES N-(2-hydroxyethyl)piperazine-N-2-ethanesulfonic acid - HPLC high-performance liquid chromatography - SDS sodium dodecyl sulfate     

Immunotherapy with bestatin for acute nonlymphocytic leukemia in adults

Summary In a cooperative randomized control study of immunotherapy with bestatin in combination with chemotherapy in adults with acute nonlymphocytic leukemia (ANLL), 101 patients (48 in the bestatin group and 53 in the control group) out of 115 patients registered were evaluated as eligible. The bestatin group achieved a statistically significant prolongation of survival compared with the control group in overall ANLL and acute myelogenous leukemia. In the analysis of patient age, the bestatin group achieved a statistically significant prolongation of both the remission duration and survival in patients aged 50 to 65 years, while the differences were not significant in the 15 to 49 age group. The bestatin group tended to achieve a higher rate of reinduction of remission in patients who had recurrence of leukemia. Side effects developed in only 5 (9.6%) of 52 patients treated with bestatin. None of these side effects were particularly serious in nature. It is concluded that bestatin is useful for prolongation of survival of adult patients with ANLL, making for a longer remission duration especially in elderly patients and with few side effects.     

Location of the inhibitory region of smooth muscle myosin light chain kinase

Proteolysis by trypsin of gizzard myosin light chain kinase (MLC kinase) in the absence of Ca2+-calmodulin produced a 64,000-dalton inactive fragment which was converted to a 61,000-dalton Ca2+-calmodulin-independent active fragment. This confirmed previous results (Ikebe, M., Stepinska, M., Kemp, B. E., Means, A. R., and Hartshorne, D. J. (1987) J. Biol. Chem. 262, 13828-13834). On the other hand, proteolysis of MLC kinase in the presence of Ca2+-calmodulin initially produced a 66,000-dalton Ca2+-calmodulin-dependent active fragment which was converted to a 61,000-dalton Ca2+-calmodulin-independent active fragment with further proteolysis. The amino acid sequences from the N terminus of the 66,000-dalton, 64,000-dalton, and 61,000-dalton fragments were determined. The sequence was not found in the reported partial amino acid sequence of MLC kinase (C-terminal 60% of whole sequence) (Guerriero, V., Jr., Russo, M. A., Olson, N. J., Putkey, J. A., and Means, A. R. (1986) Biochemistry 25, 8372-8381), and, therefore, the cleavage sites are in the remaining 40% N-terminal portion of the sequence of MLC kinase. The C terminus of these MLC kinase fragments was determined by employing the carboxypeptidases A, B, and Y digestion followed by the amino acid analysis of the released amino acids. As a result, it was concluded that the C terminus of the 66,000-dalton, 64,000-dalton, and 61,000-dalton MLC kinase fragments are arginine 522, lysine 490 and arginine 494, and lysine 473, respectively. These results show that the inhibitory domain is in the amino acid sequence of 474-490, and that the amino acid sequence 494-522 confers the calmodulin-dependent kinase activity.