Study of edema reduction patterns during the treatment phase of complex decongestive physiotherapy for extremity lymphedema.

Shortening the treatment phase of complex decongestive physiotherapy (CDP) is extremely important both for individual patients and medical economics. In 83 patients with stage II unilateral secondary extremity lymphedema (31 upper extremities and 52 lower extremities), the daily changes in the volume of affected extremities during the treatment phase of CDP were prospectively investigated. For the upper extremity lymphedemas, the biggest change was seen between days 1 (100% residual edema rate) and 2 (46.0 +/- 2.7%; mean +/- SD) of therapy with a 54.0% reduction (p < 0.0001). Between days 2 and 3 (38.0 +/- 2.6%) of therapy, there was an 8.0% reduction (p < 0.05). From days 3 to 6 of therapy, slight changes ranging from 0.2 to 3.2%/day were seen. For the lower extremity lymphedemas, the biggest change was seen between days 1 (100%) and 2 (44.5 +/- 2.1%) of therapy with a 55.5% reduction (p < 0.0001). Between days 2 and 3 (33.5 +/- 2.6%) of therapy, there was an 11.0% reduction (p < 0.001). The daily volume changes from days 4 to 6 were slight, ranging from 0.1 to 1.0%/day. During the treatment phase of CDP, the largest volume changes were seen soon after the start of therapy.

Echinococcus granulosus: Cloning and Functional in Vitro Characterization of an Actin Filament Fragmenting Protein.

We report the isolation and characterization of an Echinococcus granulosus gene that codes for a protein with actin filament fragmenting and nucleating activities (EgAFFP). The genomic region corresponding to the EgAFFP gene presents a coding sequence of 1110 bp that is interrupted by eight introns. The EgAFFP deduced amino acid sequence is about 40% homologous to those of several members of the gelsolin family, such as Physarum polycephalum fragmin, Dictyostelium discoideum severin, and Lumbricus terrestris actin modulator. As do other proteins of the same family, EgAFFP presents three repeated domains, each one characterized by internal conserved amino acid motifs. Assays with fluorescence-labeled actin showed that the full-length recombinant EgAFFP effectively binds actin monomers in both a calcium-dependent and calcium-independent manner and also presents actin nucleating and severing activities.

[Thoracoscopic pleural biopsy under local anesthesia using a 2 mm laparoscope].

Thoracoscopy is indicated in patients with undiagnosed effusion after conventional methods. It has been usually performed under general anesthesia or using a thoracoscope with a thoracoscope with a diameter over 5 mm. However, it is an invasive diagnostic technique. We evaluated the feasibility of thoracoscopic pleural biopsy under local anesthesia using a 2 mm laparoscope. Six patients with a pleural effusion of unknown etiology after conventional methods, underwent thoracoscopy under local anesthesia. A 2 mm laparoscope and biopsy forceps (2 mm Minisite, United States Surgical Corp., USA) was used in all patients. Pleural fluid was removed, and the thoracic cavity was inspected. Thoracoscopic intercostal blocks were performed with 1% lidocaine, and then a biopsy was performed. The biopsy specimen was sent for histopathology. Three patients were shown to have carcinomatous pleurisy, two of them with localized lesions less than 10 mm. In the remaining three patients, non-specific diagnoses were made, but long-term follow-up revealed no malignant pleural disease. Although the pictures obtained using a 2 mm laparoscope were inferior in quality, they were adequate for the detection of malignant lesions in the pleural cavity. There were no procedure-related complications. These findings suggest that thoracoscopy using a 2 mm laparoscope is (1) a useful diagnostic tool in cases of pleural malignancy; (2) a minimally invasive method with the advantage of being easily performed under local anesthesia. Thus, thoracoscopic pleural biopsy using a 2 mm laparoscope appears to be useful for undiagnosed pleural effusion.

Structural basis for the electron transfer across the chromaffin vesicle membranes catalyzed by cytochrome b561: analyses of cDNA nucleotide sequences and visible absorption spectra.

We isolated cDNA clones for cytochromes b561 from sheep and porcine adrenal medullae using the RT-PCR technique. Comparison of the deduced amino acid sequences of various species showed that there are two fully-conserved regions in this cytochrome. In addition, one methionyl and six histidyl residues (potential heme ligands) are fully-conserved. Based on a plausible structural model in which a polypeptide spans the vesicle membranes six times and holds two heme B molecules, the first conserved sequence (69ALLVYRVFR77) is located on the extravesicular side of an alpha-helical segment and the second one (120SLHSW124) is located in an intravesicular loop connecting two alpha-helical segments, respectively. Consideration of the relative locations of the fully-conserved sequences, and the methionyl and histidyl residues in the model led to a proposal that the first and second conserved sequences are likely to form the binding sites for extravesicular ascorbic acid and intravesicular semidehydroascorbic acid, respectively. A mild alkaline-treatment of purified bovine cytochrome b561 in oxidized state led to a specific loss of an electron-accepting ability from ascorbic acid for a half of the heme center, suggesting a distinct role for each of the two hemes.

Phenylethanoid glycosides from Stachys officinalis.

From the aerial parts of Stachys officinalis, six new phenylethanoid glycosides, named betonyosides A-F, and six known phenylethanoid glycosides, acetoside, acetoside isomer, campneosides II, forsythoside B and leucosceptoside B, were isolated and their structures were elucidated from spectroscopic and chemical evidence. Campneosides II were separated into two epimers.

Effects of hypergravity on growth and cell wall properties of cress hypocotyls.

Elongation growth of etiolated hypocotyls of cress (Lepidium sativum L.) was suppressed when they were exposed to basipetal hypergravity at 35 x g and above. Acceleration at 135 x g caused a decrease in the mechanical extensibility and an increase in the minimum stress-relaxation time of the cell wall. Such changes in the mechanical properties of the cell wall were prominent in the lower regions of hypocotyls. The amounts of cell wall polysaccharides per unit length of hypocotyls increased under the hypergravity condition and, in particular, the increase in the amount of cellulose in the lower regions was conspicuous. Hypergravity did not influence the neutral sugar composition of either the pectin or the hemicellulose fraction. The amount of lignin was also increased by hypergravity treatment, although the level was low. The data suggest that hypergravity modifies the metabolism of cell wall components and thus makes the cell wall thick and rigid, thereby inhibiting elongation growth of cress hypocotyls. These changes may contribute to the plants' ability to sustain their structures against hypergravity.

Apoptosis of acinar cells in the pancreas of rats fed on a copper-depleted diet.

Male Fischer 344 rats weighing 80-90 g were fed on a copper-depleted diet supplemented with 0.6% triethylenetetramine tetrahydrochloride (a copper chelator), and the death of pancreatic acinar cells of these rats was investigated morphologically and biochemically. The weight of the pancreas of these rats decreased from 3 weeks after feeding, and concomitantly the percentage of dead acinar cells increased to the maximum in about the 5th week and decreased subsequently. These dead acinar cells showed light microscopic and electron microscopic characteristics of apoptosis. Furthermore, the electrophoretic pattern of DNAs extracted from the pancreas having many dead acinar cells showed a ladder-like distribution, characteristic of apoptosis. The present results indicate that feeding of rats on a copper-depleted diet supplemented with a copper chelator results in apoptosis of acinar cells of the pancreas.

[Clinical experience of local thermotherapy in the treatment of benign prostatic hyperplasia].

Thirty-five patients with benign prostatic hyperplasia (age range: 45-88 years; average: 67.5 years) underwent local thermotherapy with prostathermer. Clinical therapeutic effect was evaluated in 30 of the 35 patients: 2 patients interrupting from therapy and 3 receiving pretherapeutic indwelling catheters were not included. A total of 6 treatments (2 per week) were performed, each lasting for 60 minutes. As for subjective improvement, improvement of nocturia was noted in 70.0% of all patients and sense of residual urine in 70.7%. Post-therapeutic nocturnal and daytime decreases in urination frequency were statistically significant (p less than 0.01). Objective improvement in residual urine volume occurred in 19 of the 30 cases, and elevation in uroflowmetric maximal flow rate following therapy was statistically significant (p less than 0.05). Among complications ascribable to catheter insertion were urethral bleeding (3 cases), epididymitis (1 case) and pyuria (1 case). Therapeutic result based primarily on subjective symptoms and partly on objective findings was fairly good in 17 cases (about 57%), and slightly good in 25 cases (about 83%). In conclusion, this therapy seems to be useful in the treatment of benign prostatic hyperplasia.

A computer simulation of the creep process of the cell wall using stress relaxation parameters.

The stress relaxation of the cell wall of pea plants was measured and viscoelastic parameters were obtained according to the stress-relaxation analysis developed by Yamamoto et al. (Plant & Cell Physiol. 1970). The creep process of the cell wall was simulated by a numerical integration using stress-relaxation parameters, because the direct conversion of the stress relaxation process to the creep is impractical. In the conversion, a personal computer was programmed for the Maxwell viscoelastic model with a compiler language. Cell wall creep was measured with a specially constituted apparatus and compared with that calculated by the simulation process. The results suggested that the creep can be reproduced by a computer simulation using the stress-relaxation parameters. Both creep and stress-relaxation properties of plant cell walls can be analyzed by using a single model.

Human diabetes associated with a mutation in the tyrosine kinase domain of the insulin receptor.

Insulin receptor complementary DNA has been cloned from an insulin-resistant individual whose receptors have impaired tyrosine protein kinase activity. One of this individual's alleles has a mutation in which valine is substituted for Gly996, the third glycine in the conserved Gly-X-Gly-X-X-Gly motif in the putative binding site fo adenosine triphosphate. Expression of the mutant receptor by transfection into Chinese hamster ovary cells confirmed that the mutation impairs tyrosine kinase activity.

Molecular cloning of plant transcripts encoding protein kinase homologs.

Oligonucleotides, corresponding to conserved regions of animal protein-serine/threonine kinases, were used to isolate cDNAs encoding plant homologs in the dicot bean (Phaseolus vulgaris L.) and the monocot rice (Oryzae sativa L.). The C-terminal regions of the deduced polypeptides encoded by the bean (PVPK-1) and rice (G11A) cDNAs, prepared from mRNAs of suspension cultures and leaves, respectively, contain features characteristic of the catalytic domains of eukaryotic protein-serine/threonine kinases, indicating that these cDNAs encode plant protein kinases. The putative catalytic domains are most closely related to cyclic nucleotide-dependent protein kinases and the protein kinase C family, suggesting the plant homologs may likewise transduce extracellular signals. However, outside these domains, PVPK-1 and G11A exhibit no homology either to each other or to regulatory domains of other protein kinases, indicating the plant homologs are modulated by other signals. PVPK-1 corresponds to a 2.4-kb transcript in suspension cultured bean cells. Southern blots of genomic DNA indicate that PVPK-1 and G11A correspond to single copy genes that form part of a family of related plant sequences.

Interaction of rat ascites hepatoma cells with cultured mesothelial cell layers: a model for tumor invasion.

Interactions of rat ascites hepatoma cells with primary cultured layers of rat mesentery-derived cells were studied. The mesentery-derived cells were isolated from rat mesentery and cultured in Eagle's minimum essential medium with a 2-fold concentration of amino acids and vitamins supplemented with 10% calf serum. The primary cultured cells, consisting mainly of mesothelial cells in polygonal shape, forms a "paving stone" sheet. Upon seeding the tumor cells on the mesentery-derived cell layers, three different types of tumor cell growth were observed. Type 1 was the formation of piled-up tumor cell nests on mesothelial cell layers. Type 2 was the formation of flattened tumor cell islands underneath mesothelial cell layers. This island formation was clearly observed under a phase contrast microscope 2 days after the tumor cell seeding. Protrusion of cellular processes of the tumor cells beneath mesothelial cells was occasionally seen. Type 3 was the growth of tumor cells in suspension. These types of tumor cell growth closely resemble those in the peritoneal cavity observed after i.p. implantation of the tumor cells. When the tumor cells recovered from the blood of tumor-bearing rats were seeded, flattened tumor cell islands were formed 15 times more frequently than when the tumor cells isolated from host peritoneal cavity were seeded. Shortly after the appearance of small flattened tumor cell islands, a distinct morphological change of mesothelial cells from polygonal to spindle shape was seen preferentially at the marginal area of the cell layers (a partial retraction of cell edges). The retraction of mesothelial cells was induced not only by seeding the tumor cells but by adding the tumor ascites fluid or the medium conditioned by the tumor cell culture. The morphological change was reversed by changing the culture medium to remove the effectors. These results indicate that the system described in this study can provide a useful model to study tumor cell invasion.

Elemental contamination of Japanese women's hair from historical samples.

Japanese women's hair, which had been cut in the past and preserved was examined for the presence of 12 elements (Na, K, Mg, Ca, Sr, Mn, Fe, Cu, Zn, Hg, Pb and P). Elevated levels of Fe, Mn, Cu, Pb and Hg (inorganic mercury) were conspicuous in the samples cut in the period 1880-1929 and used as hair pieces (kamoji). The effects of washing on element concentration depended on both the method of washing and the type of element. The least effective was washing with acetone and water, compared with two other methods involving anionic or non-ionic surface active agents. The most resistant elements to washing were Pb, Hg, Cu and Zn. From the intercorrelation of element content and factor analysis, by examining the diminution of contents by washing and by comparing the detected levels with the values measured on contemporary Japanese women's hair, the contribution of exogenous contamination to hair levels was found to be very strong for Fe, Mn, Cu, Hg (inorganic mercury), and Pb, moderate for Na and Zn, and negligible for Ca, Mg, Sr, K, Hg (organic mercury), and P.

Alteration of inorganic mercury accumulation due to selenite in organs of mice fed methylmercury.

Mice were fed methylmercury (10 nmol/g feed) and selenite (0, 8, 20 or 50 nmol/ml drinking water) for one or two weeks. Doses of selenite and duration of feeding were determining factors of total mercury and inorganic mercury concentrations in organs. Increasing the dose of selenite produced the following results: concentration of total mercury increased in the brain and liver and decreased in the blood, kidneys and spleen; concentration of inorganic mercury increased in the liver and spleen, decreased in the kidneys, and remained unchanged in the brain; the rate of inorganic mercury to total mercury increased in the liver and spleen, decreased in the brain, and remained unchanged in the kidneys. In every case, inorganic mercury in the blood was below the detection limit.

Effects of methylmercury and selenite on osmolarity and electrolytes in blood of mice.

Female IVCS mice were fed methylmercury at a low dose (10 nmol/g feed) with each of the doses of selenite (0, 8, 20 and 50 nmol/ml of drinking water) for 2 weeks. Mice fed methylmercury alone showed increased osmolarity of plasma: coadministration of selenite produced no change even at the lowest dose. Selenite alone did not change the osmolarity. Both concentrations of Na+ and total protein in plasma increased due to administration of methylmercury alone without change of K+ and Cl- levels. Coadministered selenite abolished this effect of methylmercury on the electrolytes and total protein of plasma.

Decreased membrane fragility of mouse erythrocytes by small dose of methylmercury and its restoration by coadministered selenite.

Female IVCS mice were fed methylmercury at a low dose level (10 nmoles/g feed) with coadministration of selenite (0, 8, 20 or 50 nmoles/ml of drinking water) for one week or two weeks. Mice fed methylmercury alone showed the decreased fragility of erythrocytes membrane when compared with the non-treated control. Coadministration of selenite produced such a change to smaller extents than methylmercury treatment alone. Mercury levels in the blood varied according to the dose level of selenite (the highest level was found in methylmercury alone group.)

Efficacy of Linco-Spectin medication on mycoplasma meleagridis airsacculitis in turkey poults.

The efficacy of Linco-Spectin (LS) water medication was determined against Mycoplasma meleagridis (MM) airsacculitis in turkey poults under controlled conditions. The poults were obtained from commercial flocks naturally infected with MM. Water medication was given for the first five days of life. In Georgia and Ohio, LS was given at 0, 1, 2, and 4 g per gallon of drinking water. In California and Minnesota, LS was administered at 0 and 2 g per gallon of drinking water. At three weeks old, the birds were weighed individually and examined serologically, culturally, and grossly for MM airsacculitis. LS at 2 g activity per gallon of drinking water was beneficial in controlling MM airsacculitis in all four trials.