Subunit-specific sulphation of oligosaccharides relating to charge-heterogeneity in porcine lutrophin isoforms.

Lutrophin (LH) consists of an array of isoforms with different charges and bioactivities. This study was undertaken to clarify specifically how oligosaccharides of alpha and beta subunits contribute to LH isoform charges. Porcine LH (pLH) was separated into four isoforms by isoelectric focusing (IEF), followed by subunit isolation. Their oligosaccharides were released by hydrazinolysis, labelled by reduction with NaB3H4, and fractionated by HPLC with a Mono Q column into five populations differing in the number of sulphate (S) and sialic acid (N) residues, designated as Neutral, N-1, S-1, S-N and S-2. Oligosaccharides were predominantly sulphated (S-1 and S-2) and infrequently sialylated (N-1 and S-N). Further analysis, including concanavalin A (Con A) affinity chromatography, desialylation, desulphation, sequential exoglycosidase digestion and methylation, clarified the structures of the acidic oligosaccharides. All were of the biantennary complex type. Their two peripheral branches were SO4-4GalNAc beta 1-4Glc-NAc and GalNAc beta 1-4GlcNAc or GlcNAc in S-1, SO4-4GalNAc beta 1-4GlcNAc and Sia alpha 2-6Gal beta 1-4GlcNAc in S-N, and (SO4-4GalNAc beta 1-4GlcNAc)2 in S-2 (where GalNAc is N-acetylgalactosamine and GlcNAc is N-acetylglucosamine). Ten percent of S-1 and of S-N had a bisecting GlcNAc residue. Sulphate residues occurred in nearly the same amount for both subunits; however, the alpha and beta subunits were sulphated differently. S-1 predominated in the alpha subunit, while S-1 and S-2 were major components in the beta subunit.(ABSTRACT TRUNCATED AT 250 WORDS)


Structural analysis of the N-linked glycan chains from a stylar glycoprotein associated with expression of self-incompatibility in Nicotiana alata.

Self-incompatibility in flowering plants of the family Solanaceae is mediated by the product of the S-allele. The allelic products of the S-gene in the female sexual tissues of the pistil are glycoproteins in the mol. wt range 28-32 kDa. These S-glycoproteins have been isolated from styles of Nicotiana alata, homozygous for the S1- and S2-alleles. Earlier studies have indicated that the single potential N-glycosylation site on the S1-glycoprotein bears a glycan chain, whereas of the four potential N-glycosylation sites on the S2-glycoprotein, three are glycosylated. This paper describes the purification and characterization of the N-linked glycan chains from these two glycoproteins. Oligosaccharides were cleaved off the glycoproteins using peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase F (N-glycanase F) and separated by anion-exchange HPLC. Four types of hybrid structure were defined by chemical techniques, fast atom bombardment-mass spectrometry (FAB-MS) and 1H-NMR. Although the relative amounts differed, all four structures were found on both the S1- and S2-glycoproteins, and are heterogeneous at some N-glycosylation sites. No O-linked glycans were detected on the S2-glycoprotein. These results are discussed in relation to the potential of the structural diversity residing in this array of glycoforms to play a rôle in allelic specificity.


Synthesis of a photoreactive, radiolabelled derivative of the oligosaccharide of GM1 ganglioside.

A photoreactive, radioiodinatable derivative of the oligosaccharide (GM1OS) of ganglioside GM1 was synthesized as follows: GM1OS was generated from GM1 by ozonolysis and alkaline fragmentation, and reductively aminated to GM1OSNH2 (1-amino-1-deoxymonosialogangliotetraitol). The latter compound was then reacted with N-hydroxysuccinimidyl-4-azidosalicylic acid (NHS-ASA) to form GM1OSNH-ASA [1-(4-azidosalicoylamido)-1-deoxymonosialogangliotetraitol], which was radioiodinated and further purified. To test the [125I]GM1OSNH-IASA [1-(4-iodoazidosalicoylamido)-1-deoxymonosialogangliotetraitol+ ++] as a probe for ganglioside-binding proteins, the derivative was incubated with cholera toxin, which specifically binds GM1, followed by photolysis and sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The probe only labelled the B or binding subunit of cholera toxin, but not the A or adenylyl cyclase activating subunit. Labelling was inhibited by excess GM1OS, but not by the oligosaccharides from gangliosides GD1a and GD1b. [125I]GM1OSNH-IASA and analogous oligosaccharide derivatives may be valuable probes for detecting ganglioside-binding proteins.


Analysis of kidney mRNAs expressed from the rat beta-galactoside alpha 2,6-sialyltransferase gene.

beta-Galactoside alpha 2,6-sialyltransferase (SiaT-1), like other glycosyltransferases, is differentially expressed in rat tissues. Two distinct size classes of SiaT-1 mRNAs expressed in rat kidney are comprised of at least three SiaT-1 transcripts. One mRNA, RKE, represents the larger transcript class (4.7 kb) and predicts a polypeptide identical to the hepatic SiaT-1. In transfected Chinese hamster ovary (CHO) cells, RKE polypeptides exhibit hemi-perinuclear staining with a SiaT-1 antibody (Ab-267) that is consistent with Golgi localization. RKE transfectants display cell-surface alpha 2,6-sialic acid linkages as determined by lectin affinity staining. Two other mRNAs, RKA and RKB, are members of a smaller size class (3.6 kb) that comprise predominant SiaT-1 transcripts in rat kidney. Both RKA and RKB encode polypeptides that are missing the amino-terminal 232 residues, but retain 171 amino acids of RKE carboxy-terminal sequence information. A short, leucine-rich peptide present in the divergent amino-terminus of RKA has sequence similarity to the secretory signal domain of several eukaryotic secretory and cell-surface proteins. In transfected CHO cells, both RKA and RKB polypeptides display an immunostaining pattern that is distinct from that of the Golgi-associated SiaT-1 protein (RKE). Furthermore, RKA or RKB transfectants do not display alpha 2,6-sialic acid linkages on cell-surface glycoconjugates. Consistent with the expression of divergent SiaT-1 mRNAs in rat kidney, protein blot analysis of rat tissue homogenates with Ab-267 reveals that in addition to protein that co-migrates with hepatic SiaT-1, rat kidney expresses a unique size class of SiaT-1 proteins.


Model of the dynamics of receptor potential in a mechanoreceptor.

Mechanoreceptors are physiological units that convert mechanical stimuli to neural responses. An important one in the skin is the Pacinian corpuscle, which consists of a nerve ending surrounded by a capsule that is made up of alternating layers of fluid and elastic shells. In this paper a capsule model is coupled to a model for the dendrite via a variable representing the hoop strain imposed by the capsule on the receptor membrane of the dendrite. The transient behavior of the receptor potential due to step and periodic displacement-type stimuli imposed on the outer membrane of the capsule is investigated, and the results are compared to those of analogous experiments. The model is used to develop hypotheses concerning the role of the capsule as a filter for mechanical stimuli.


Computer modeling of pole formation in cell division.

The regular position of spindle poles in bi- and multipolar cell divisions is interpreted as the result of expanding caps on the surface of cell nuclei first proposed by Mazia in 1986. In a computer model it is shown that expanding caps position their centers with the maximal distance between them, which is in complete accordance with findings in experimentally influenced cell divisions in which multiple poles are formed.


Estimation of parity progression ratios from the truncated distribution of closed and open birth intervals.

A procedure to estimate parity progression ratios in a population from the truncated distribution of open and closed birth intervals is presented. The approach is quite simple in computation and data needs. It does not require any separate data on age at last birth to the women of completed fertility as in earlier methods. The procedure is illustrated with an observed set of data.


A method for the analysis of biological transduction phenomena.

Biological transduction can be defined as the triggering of a cellular response by the binding of molecules of effector substances to specific cellular sites. An example of biological transduction, analyzed in this report, is the triggering of T-cell proliferation by the binding of T-cell growth factor (TCGF) to specific TCGF-binding sites on responsive T-cells. Sigmoidal or S-shaped curves often result when measurements of biological response are plotted as a function of concentration of effector substance. Such curves suggest that effector molecules must bind a critical number of cellular sites, and this critical number of bound complexes must undergo secondary events (cross-linking, association, internalization, second messenger release, etc.) in order to initiate the biological response. The method described here estimates the critical number of cellular sites (R) and the probability of these secondary events (PS/B) as follows: (1) The total number of cellular sites (N) is estimated from binding data, and the probabilities (PB) of effector molecules binding to a site are estimated from response data. (2) The response data are assumed to follow the summed binomial distribution function, which is equated to the incomplete beta function. (3) R and PS/B are estimated by applying nonlinear regression to the incomplete beta function. The T-cell data to which the method was applied gave N = 15,000, R = 5, and PS/B = 7.22 x 10(-4). These results show that the binding of very few TCGF molecules is required for activation of T-cells and that the probability of the secondary events leading to cell proliferation is much smaller than the probability of TCGF binding to T-cells. The method described can be used to analyze any biological transduction experiments where both binding and biological response data are available.


Optimal control of tumor size used to maximize survival time when cells are resistant to chemotherapy.

The high failure rates encountered in the chemotherapy of some cancers suggest that drug resistance is a common phenomenon. In the current study, the tumor burden during therapy is used to slow the growth of the drug-resistant cells, thereby maximizing the survival time of the host. Three types of tumor growth model are investigated--Gompertz, logistic, and exponential. For each model, feedback controls are constructed that specify the optimal tumor mass as a function of the size of the resistant subpopulation. For exponential and logistic tumor growth, the tumor burden during therapy is shown to have little impact upon survival time. When the tumor is in Gompertz growth, therapies maintaining a large tumor burden double and sometimes triple the survival time under aggressive therapies. Aggressive therapies aim for a rapid reduction in the sensitive cell subpopulation. These conclusions are not dependent upon the values of the model constants that determine the mass of resistant cells. Since treatments maintaining a high tumor burden are optimal for Gompertz tumor growth and close to optimal for exponential and logistic tumor growth, it may no longer be necessary to know the growth characteristics of a tumor to schedule anticancer drugs.


Low-intensity combination chemotherapy maximizes host survival time for tumors containing drug-resistant cells.

Recent clinical trials have shown that for some cancers, high-intensity alternating chemotherapy does not significantly improve either survival times or response rates compared with nonalternating therapy. The current study uses optimal control to determine the best way to treat a tumor that contains drug-resistant cells that cannot be destroyed. The delivery of two non-cross-resistant chemotherapeutic agents is limited by bounds on the drug concentration and the dose intensity. This ensures that the drug toxicity stays within a tolerable range. The aim of the therapy is to maximize the host survival time, defined as the time over which the tumor burden can be kept below a fixed bound. The model is posed as a free terminal time, optimal parameter selection problem in which the constraints are continuously parametrized by time and the number of courses of therapy is free to vary. New theory is developed so that the optimal parameter selection problem can be solved as a sequence of fixed terminal time problems using existing optimal control software. Numerical simulations of Gompertz tumor growth showed that a treatment maintaining a high tumor burden doubled and sometimes tripled with survival time under aggressive therapy. When these simulations were repeated using exponential and logistic tumor growth models, the tumor burden during treatment had little influence upon survival time. In all simulations, survival time was not extended by delivering the anticancer drugs concurrently instead of staggering the treatment arms.


Maximum sustainable yield of populations with continuous age-structure.

We address the problem of finding the harvesting policy that will maximize the yield and maintain a population in a steady state. The population is characterized by continuous age classes and therefore follows differential equations. Here, we assume that the equations are linear (no density dependence). Two possible constraints are considered: either recruitment or total population are fixed to a constant. Under these conditions, the optimal policy is to harvest the fraction theta of a younger age class ã and to harvest totally an older age class b. The optimal solution (theta, ã, b) can be calculated explicitly if the fecundity and mortality schedules are given. The solution is compared to the simpler strategy of harvesting all individuals beyond a single age class â. It is shown that the latter strategy can be much less profitable than harvesting two age classes because it cannot take account of the different values of individuals according to their age.


Enteral nutrition: accessing patients.

This review highlights important points in regard to use of the enteral route to provide nutrition support to nutritionally compromised patients. Enteral nutrition has become technologically advanced medical therapy in the last 15 yr, and as clinical nutrition continues to develop, further advances will be made. With the range of access routes and delivery systems available, appropriate nutrition can be provided in a safe, effective manner.


Catabolic response and parenteral nutrition after simultaneous kidney-pancreas transplantation.

The aim of the study was to quantify the catabolism rate induced by simultaneous kidney-pancreas transplantation and to evaluate the impact of parenteral nutrition (PN) on recovery of graft function. Twenty-six diabetic uremic patients were studied. The average urea nitrogen production (UNP) was 5.2 +/- 1.7 g during the first 24 h after transplantation, while patients did not receive energy and nitrogen support. Energy (30 kcal.kg-1.day-1) and nitrogen (0.15 g.kg-1.day-1) intake started 24 h after surgery. In 14 patients, a mixed regimen was adopted (70% carbohydrates, 30% lipids), and 12 patients received only hypertonic glycidic solutions. The recovery of kidney function was immediate in all cases, with a prompt decrease in blood urea nitrogen and serum creatinine levels. C-peptide levels rose immediately after the revascularization of the pancreas graft and remained within the normal range during the PN period. No significant difference was observed in UNP or glucose tolerance between the mixed-regimen and glycidic groups. However, on average 6.6, and 1.5 hyperglycemic episodes occurred during the 1st wk of PN in the glycidic and mixed-regimen groups, respectively. The posttransplantation catabolism rate was similar to that induced by an elective major surgical procedure. Eucaloric PN did not affect the recovery of kidney and pancreas graft function. A mixed energy regimen seems to be most suitable for kidney-pancreas transplant patients because it prevents hyperglycemia which might be misdiagnosed as rejection.


Variations in plasma amino acids in septic patients subjected to parenteral nutrition with a high proportion of branched-chain amino acids.

Sepsis is characterized by an increase in the plasma concentration of aromatic amino acids (AAAs) and those containing sulfur and a decrease in the branched-chain amino acids (BCAAs). We studied changes in the plasma aminogram of septic patients given different types of total parenteral nutrition (TPN), analyzing variations in accordance with the type of TPN used and the importance that the use of BCAA may have in these patients. We studied 80 patients with peritonitis divided into two groups of 40 patients each: group 1 was given a solution with 22.5% BCAA and group 2 a solution with 45% BCAA. High BCAA content caused an increase in the plasma concentrations of these amino acids and in the BCAA/AAA quotient and a decrease in AAAs. Plasma concentrations of leucine and valine reached high, potentially toxic levels at 15 days when solutions with high BCAA content were used. Glycine increased in group 1, which may be important because of its tendency to produce hyperammonemia. BCAAs are of unquestioned nutritional importance in view of the evidence of changes that take place in muscle protein catabolism and in plasma amino acids. In the phase of increased protein catabolism, we saw a plasma amino acid pattern in keeping with the existing metabolic situation. The need for BCAA diminishes when the hypercatabolic state disappears.


Glucose metabolism in advanced lung cancer patients.

Although it is generally accepted that altered nutrient intake and metabolism are responsible for the progressive loss of body weight observed in most advanced cancer patients, there is still considerable controversy regarding the contributory role of changes in both resting energy expenditure (REE) and glucose metabolism. Several studies suggest increases in both REE and glucose appearance in advanced cancer patients compared with healthy control subjects, whereas others revealed no changes in either metabolic parameter. We measured REE with indirect calorimetry and glucose kinetics with a primed constant infusion of D-[U-14C]glucose and D-[6-3H]glucose over the last 4 h of a 24-h fast in 32 advanced lung cancer patients immediately after diagnosis and before any chemotherapy or radiotherapy and in 19 healthy volunteer subjects. REE for the lung cancer group was not significantly different from that in the control group (1535.8 +/- 78.0 vs. 1670.2 +/- 53.9 kcal/day, respectively, p = 0.151). When REE was expressed as a function of body weight, or lean body mass, no differences between the two groups were observed. The rate of glucose appearance was 9.88 +/- 0.36 mumol.kg-1.min-1 in the cancer patients and 10.15 +/- 0.53 mumol.kg-1.min-1 in control subjects (p = 0.667), of which 50.4 versus 58.2%, respectively, was oxidized. The amount of glucose recycled was 13.54 +/- 1.22% in cancer patients and 15.08 +/- 0.99% in control subjects (p = 0.394). The amount of VCO2 from direct oxidation of glucose was 23.39 +/- 0.74% in cancer patients and 27.45 +/- 1.36% in control subjects (p = 0.006).(ABSTRACT TRUNCATED AT 250 WORDS)


Total dietary restriction and thymus, spleen, and phenotype and function of splenocytes in growing mice.

To study the immunological effect of total dietary restriction, BALB/c mice were limited to reduced body growth or maintained at practically constant body weight between 21 and 61 days of age by giving them 70% (R70%) or 55% (R55%) of the average daily food intake of the control group fed ad libitum. Thymus and spleen weight were decreased, but thymus size was maintained in proportion to body weight in R70% mice, whereas the ratio of thymus weight to body weight was significantly decreased in R55% mice. In restricted mice, splenocytes showed a lower percentage of B lymphocytes and a higher percentage of T lymphocytes. Results of stimulation showed that proliferation capacity was increased for B lymphocytes and decreased for T lymphocytes in restricted mice. Our data show the importance of studying the thymus over a period, because normal thymus growth was first greatly impaired by dietary restriction, and subsequent thymus involution was delayed, showing that an isolated comparison between the thymus of growing mice is not enough. Our data show the interest of determining the percentage of lymphocyte populations parallel to their response to mitogen stimulation.


Effects of graded levels of high-molecular-weight carrageenan on colonic mucosal thymidine kinase activity.

Graded levels of high-molecular-weight carrageenan were fed to male Fischer 344 rats to investigate the effects of carrageenan on colonic cell proliferation. Four groups of 7 rats consumed diets containing 0 (control), 0.65, 1.31, or 2.61% carrageenan ad libitum for 4 wk. These levels were designed to simulate (on a milligram-per-kilogram basis) 25, 50, and 100 times the maximal human carrageenan intake. After an overnight fast, the rats were anesthetized, and the colonic mucosa was removed and homogenized. A supernatant was prepared and assayed for total protein and thymidine kinase activity (a marker for cell proliferation). Increasing levels of carrageenan resulted in stepwise increases in total mucosal protein and in thymidine kinase specific (units per milligram of protein) and total (units per centimeter of colon) activity. However, increases in thymidine kinase activity were statistically significant only at the highest dose of carrageenan. No histological abnormalities were associated with any level of carrageenan feeding. These results suggest a clear dose-response increase in colonic mucosal cell proliferation with increasing doses of carrageenan; however, the effect at doses approximating 19 times maximal human intake was not statistically different from that for control rats under the conditions of this study.


Total parenteral nutrition in surgical illness: how much? How good?

Using primed constant infusions of isotopes and indirect calorimetry, we assessed protein, glucose, and fat kinetics in severely ill surgical patients suffering from sepsis, major trauma, gastrointestinal cancer, or nutritional depletion from benign disease. We also assessed the effect of 5 days of total parenteral nutrition (TPN) on abnormal metabolism in these states. In the basal state, patients with sepsis, trauma, or cancer had an elevated rate of net protein catabolism due to an increased rate of whole-body protein catabolism, whereas whole-body protein synthesis was not impaired. TPN had no impact on the elevated rate of whole-body protein catabolism in these conditions but decreased the rate of net protein catabolism, suggesting that increased substrate availability optimizes whole-body protein synthesis. Consequently, few surgical patients became anabolic while receiving TPN. In contrast, patients with nutritional depletion from benign disease had a decreased rate of net protein loss compared with volunteers and could be made anabolic with the administration of TPN. All patients studied had an elevated rate of plasma glucose production, impaired glucose oxidation, and an increased rate of Cori cycling in the basal state. After 5 days of TPN, most surgical patients showed improved ability to oxidize glucose, but the high rates of glucose recycling to lactate persisted. Patients suffering from sepsis, trauma, or cancer had an enhanced rate of lipolysis in the basal state associated with an increased rate of whole body-fat oxidation compared with healthy volunteers. After administration of TPN, whole-body fat oxidation was significantly decreased in patients with trauma but increased in patients with sepsis or cancer.(ABSTRACT TRUNCATED AT 250 WORDS)


The relationship of estrogen receptor status to DNA ploidy in breast cancer.

The relationship of estrogen receptor (ER) status to DNA ploidy was investigated in 121 patients with breast cancer who underwent surgery. Lymph node status was evaluated histologically and ER levels were determined by the dextran-coated charcoal method, with a level of 3 fmol/mg.protein being considered positive. Flow cytometric DNA content was analyzed using paraffin-embedded tissue blocks. Sixty-three percent of the specimens were ER+, while 37 percent were negative. Sixty-one patients (50.4 percent) were diploid and 60 aneuploid. A statistically significant correlation between the ER positivity rate and diploid DNA was found. Higher ER levels were seen in the postmenopausal patients with diploid tumors than in those with aneuploid tumors and there was a significant tendency for ER levels to be higher in the diploid tumors. Nodal status was not correlated with ER positivity or ploidy pattern. The present results indicate that ER levels are correlated with DNA ploidy, and reflect the degree of functional differentiation.


Postoperative pyothorax.

Twenty-five cases of pyothorax occurred in a series of 1281 thoracotomies. Almost all cases of pyothorax without bronchopleural fistula were successfully treated by closed drainage and irrigation alone. On the other hand, patients with pyothorax and fistula who were treated only with closed drainage almost all had a poor outcome. When pyothorax with fistula was treated by closed drainage and irrigation followed by further procedures such as open window thoracostomy, muscle plombage and/or omentopexy, treatment was successful. It is concluded that pyothorax without fistula may be cured by closed drainage and irrigation alone, but that pyothorax with fistula requires operative intervention such as open window thoracostomy or omental flap as soon as possible.


Radical mastectomy with parasternal node dissection or radiation to the parasternal region for breast cancer of medial or central location.

In order to establish a therapeutic approach for primary breast cancer of medial and central origin, we reviewed 183 patients who had been treated by one of the following three modalities at the Second Department of Surgery, Osaka University Medical School between January, 1965 and December, 1980. Group A (n = 70): standard radical mastectomy alone; Group B (n = 34): standard radical mastectomy followed by postoperative irradiation to the parasternal and supraclavicular regions, and; Group C (n = 62): extended radical mastectomy that included removal of the parasternal lymph nodes. The background factors of the three groups were not significantly different. The overall survival five and ten years following surgery in the three groups were 91 percent and 79 percent in group A, 82 percent and 67 percent in group B, and 82 percent and 70 percent in group C, respectively, showing no significant difference in overall survival among the three groups. When the patients were classified according to the extent of axillary lymph node involvement, there was no difference in survival among the three treatments in patients who had less than three lymph node metastases in the axilla. However, treatment of the parasternal lymph nodes improved survival in the patients who had more than four lymph node metastases in the axilla. Parasternal lymph node involvement definitely worsened the prognosis, showing it to be a good prognostic factor. Thus, extended radical mastectomy should be considered for patients with breast cancer of medial or central location, when extended axillary lymph node involvement is found.


The role of the abdominal sympathetic nervous system in regulating portal venous flow and its functional distribution.

The role of the abdominal sympathetic nervous system in regulating portal venous flow (PVF) was examined in anesthetized rats using an ultrasonic volume flowmeter. Electrical stimulation of the hepatic sympathetic branch, given at 10 Hz, 1 ms, 10 s and 12 V, caused approximately a 28 percent reduction in PVF, which was equivalent to that produced by occlusion of the bilateral carotid arteries for 30 s, without causing any change in the systemic arterial pressure. Stimulation of the major splanchnic nerve decreased PVF, the response being greater by stimulation of the right nerve than by stimulation of the left (p less than 0.05). Bilateral adrenalectomy shortened the recovery time without changing the magnitude or lateral predominancy. Neither proper hepatic arterial occlusion nor partial hepatectomy affected the response. In the partially hepatectomized animals, stimulation of the hepatic branch did not decrease the splenic flow but decreased the superior mesenteric venous flow (SMVF) and induced a similar response in PVF even when the SMVF was interrupted. An intraportal injection of noradrenaline decreased PVF dose-dependently. These findings indicate that the sympathetic nerve regulates PVF directly, and that there is functional laterality of the regulatory mechanism in the abdominal cavity, which suggests that adrenal factors work together with the nerve that supplies the portal vein.


A novel anticancer treatment for xenoplanted human gastric cancer using polyamine antimetabolites in a low polyamine diet.

The aim of the present study was to evaluate a new anticancer treatment for gastrointestinal cancer, using a combination of polyamine antimetabolites, an anticancer agent and a low-polyamine state. Two polyamine antimetabolites, given as either 40 mg/kg of methylglyoxal-bis-guanylhydrazone (MGBG) or ethylglyoxal-bis-guanylhydrazone (EGBG) and a normal diet (ND), or 20 mg/kg of each drug and a low polyamine diet (LPD), together with 1,000 mg/kg of alphadifluoromethylornithine (DFMO) were administered ip to nude mice for six consecutive days. Mitomycin C (MMC) at 2 mg/kg was then given ip for 3 alternate days. The combination of MGBG or EGBG with DFMO plus MMC resulted in an enhanced antitumor efficacy on LPD. However, the combination which included EGBG was much more enhanced than that which included MGBG and there was no evidence of any tumor regrowth. Weight loss was minimal or nil in the mice given the combination with EGBG, but was evident in those given the combination with MGBG. These results led to the conclusion that in mice, the combined therapy of EGBG with DFMO plus MMC and LPD is a safe and effective regimen for the treatment of gastric cancer.


Portal circulation and the function of hepatocytes during ex vivo perfusion of the rat liver preserved for six hours after core cooling.

Portal circulation and the function of hepatocytes during isolated organ perfusion were compared between the rat liver perfused immediately after extirpation and the liver perfused after core cooling and six hours of preservation. Nine rat livers were extirpated after core cooling, preserved for six hours in University of Wisconsin (UW) solution at 4 degrees C and then were connected to a perfusion chamber (hypothermic preservation group: 6-hr HP group). Six rat livers were immediately connected to the perfusion chamber after extirpation (control group). During 60 minutes of isolated liver perfusion, both portal circulation and the function of hepatocytes were determined every 10 minutes. Portal vein resistance increased and portal blood flow decreased during the first 20 minutes of perfusion and then stabilized in both groups. Portal vein resistance was significantly higher and portal blood flow was significantly lower for the first 10 minutes of perfusion in the 6-hr HP group, compared to the control group. The function of hepatocytes, determined by the rate of elimination of ammonia, oxygen consumption, and glucose availability were depressed in the 6-hr HP group, compared to the control groups during isolated liver perfusion. However, there was no statistically significant difference of portal perfusion and the function of hepatocytes between the two groups at the end of perfusion. The results of the present study indicate that the rat liver can be preserved in a satisfactory condition for six hours by the use of hypothermic preservation, coupled with core cooling.


Effect of prostacycline analogue OP-41483 and perfusate on liver preservation by simple hypothermia in rats.

Changes in rat livers preserved by simple cooling immersion in modified Sacks' solution, Ross' solution and Ross' solution with OP-41483, a PGI2 analogue, were compared after 12, 24, 36, and 48 hours. The viability of the preserved livers was assessed by measuring the tissue ATP concentrations and quantifying morphologic changes in the mitochondria observed by transmission electron microscopy using a 4-point scale, the mitochondrial score. A general trend towards progressive mitochondrial degeneration was observed in all groups. After 12 hrs of preservation, there was no significant difference between the groups. From 24 to 48 hours, the mitochondrial scores for livers preserved with OP-41483 were significantly higher than in the two other groups. No difference among the groups in tissue ATP concentration was observed at any time. Thus, OP-41483 was considered to retard mitochondrial degeneration in liver preservation by simple hypothermic immersion and does so by a mechanism not directly related to intracellular energy stores. The possibility that OP-41483 acts by stabilizing the lysosomal membrane is discussed.


Biodistribution of radiolabeled monoclonal antibody after intraperitoneal administration in nude mice with hepatic metastasis from human colon cancer.

The utility of the intraperitoneal (IP) administration of radiolabeled monoclonal antibody (mAb) for hepatic metastasis from human colon cancer was evaluated in congenital athymic mice. The intrasplenic injection of HT-29 LMM metastatic human colon cancer cell line reproducibly results in hepatic metastasis formation in nude mice. HT-29-15, a murine mAb of IgG1 class reactive with the HT-29 LMM cell line was labeled with iodine 125. One microgram/2 microCi of labeled HT-29-15 was injected intraperitoneally into mice with hepatic metastases, and additionally IP administration of the same dose of I-125 labeled HT-29-15 with increased volume and intravenous (IV) administration of dose quantity of HT-29-15 were performed. Blood samples were obtained at 1, 3, 5 hours, and the animals were sacrificed on days 1, 3, and 5. The percent of injected dose per gram (%ID/g) of blood after IP administration of I-125 labeled HT-29-15 reached the same level of %ID/g after IV administration by 5 hours. The transfer from the peritoneal cavity to blood was delayed by increasing the volume injected. From day 1 to day 3, there was a progressive increase for hepatic metastasis/blood ratios of I-125 labeled HT-29-15 in each group. There was no difference in the hepatic metastasis/blood ratios among the three groups. IP administration of specific mAb, therefore, provides the same level of tumor uptake in hepatic metastasis from colorectal cancer, and would be advantageous in patients with both hepatic metastasis and peritoneal implants in which radioimmunodetection and radioimmunotherapy are appropriate.


A case of linitis plastica of the rectum treated by pelvic exenteration after aggressive immunochemotherapy.

Multidisciplinary treatment was administered to a 32 year-old man with primary linitis plastica of the rectum, which was considered inoperable due to an extensive local spread at the first operation. Thirty KE of OK-432 was injected into the tumor per week (total 90-KE) and 5 KE of OK-432 was inoculated into the intracutaneous space per week (total 125 KE). Also methotrexete (MTX 50 mg) and 5-FU (500 mg) combined were given each week by intra-arterial infusion (total 20 courses). After this immuno-chemotherapy the CEA decreased from the pretreatment value of 12.7 ng/ml to 3.6 ng/ml and the tumor size was reduced. A total pelvic exenteration could then be performed almost curatively. As a result, this patient was able to return to the society with a better quality of life and has survived for 20 months since presentation.


A rare case of fibroadenoma in a tubular adenoma of the breast.

We report herein a case of a 31 year old housewife who underwent an excisional biopsy for a well-circumscribed lump in her breast. The lump contained 2 histological patterns, namely, fibroadenoma and tubular adenoma. These patterns had no transitional zone, were distinct and changed abruptly. This case was histologically diagnosed as "a fibroadenoma in tubular adenoma of the breast, benign", with no other such case ever having been reported in Japan. The histological findings of this case, convinced us that tubular adenoma is closely related to fibroadenoma.


Modified Van Praagh's operation for interrupted aortic arch with severe subaortic stenosis in a neonate.

A 26 day old neonate with Type B interrupted aortic arch, a ventricular septal defect and severe subaortic stenosis underwent a successful modified Van Praagh's operation, consisting of insertion of a graft between the main pulmonary artery and descending aorta, ligation of the patent ductus arteriosus and main pulmonary artery banding distal to the graft. This palliative procedure was effective and easily performed without the aid of cardiopulmonary bypass even in this severely ill neonate with such a complex defect.


Primary anorectal malignant melanoma.

A case of primary anorectal malignant melanoma seen in a 46 year old woman is presented herein. Her most marked symptoms were bloody stools and anal pain. Endoscopic examination indicated a tumor with ulceration but without pigmentation in the anorectal region. Histologic examination of the biopsied specimens showed spindle-shaped cells with atypia proliferating in a bandlike arrangement, as in leiomyosarcoma. An abdominoperineal resection was done and detailed histological examination of the tumor confirmed the nature of the tumor to be malignant melanoma. The postoperative immunochemotherapy consisted of Dimetyl-Triazeno-Imidasole-Carboxamide (DTIC), Amino-Methyl-Pyrimidinyl-Methyl-Chlorethyl-Nitrosourea-Hydrochlori de (ACNU), Vincristine (VCR) and OK-432. The patient has been well without recurrence for fifteen months following her operation through the continuous administration of these agents.


Surgical management of the coronary artery to pulmonary artery fistulas; a case of a large ruptured aneurysm.

Three patients were treated for coronary artery-pulmonary artery fistula. Each was asymptomatic due to a coronary artery fistula. There was one instance each of myocardial infarction, mitral stenosis and a large closed ruptured aneurysm including a thrombus. All the fistulas were comprised of several small plexiform arranged vessels. The left to right shunt ratio was approximately 8 percent or less. A surgical operation was performed to close the drainage orifice of the fistulas through pulmonary arteriotomy under cardiopulmonary bypass in two patients while one patient underwent a ligation of a large abnormal vessel to the aneurysm followed by a resection of the aneurysm without cardiopulmonary bypass. A large saccular aneurysm of such fistulas is rare and a rupture of such an aneurysm is even more rare. The surgical management of coronary artery fistulas is also discussed herein.


Multiple colorectal carcinomas and colorectal carcinoma associated with extracolonic malignancies.

In this study, we analyzed 149 surgical cases of colorectal cancer between January 1983 and August 1989. Thirteen cases (8.7 percent) of colorectal primary cancer associated with extracolonic primary malignancy of 14 lesions and 10 cases (6.7 percent) of multiple primary colorectal cancers were included. Among the 14 lesions of extracolonic primary malignancy, there were 6 gastric carcinomas, 2 endometrial carcinomas, 2 urinary bladder carcinomas, and one each in the esophagus, liver, bile duct and jejunum. The second tumor was not detected preoperatively in 3 of 4 cases of synchronous multiple primary colorectal carcinoma. A curative resection was done in 10 (77 percent) out of 13 cases of colorectal cancer associated with extracolonic malignancy, while 7 (88 percent) out of 8 cases of multiple colorectal cancers had a curative resection. Nine patients (69 percent) with colorectal cancer associated with extracolonic malignancy were disease-free for 2 months to 14 years. Seven patients (88 percent) with multiple colorectal cancers were disease-free for one to 22 years. We recommend, therefore, that in any patient with colorectal cancer, the entire large bowel should be thoroughly searched for any other primary tumors, by taking the existence of extracolonic tumors into account. A curative resection should be performed, and the follow-up period should be life-long.


Immunogenicity and immunomodulatory activity of bovine surfactant (SF-RI 1).

Respiratory distress syndrome in preterm infants can be treated successfully by endotracheal administration of a bovine surfactant preparation (SF-RI 1). Before the routine use of xenogenic surfactant preparations can be recommended, their immunogenicity as well as their in-vivo and in-vitro immunomodulatory activity have to be investigated. High titers of anti-surfactant antibodies were detected by a sensitive ELISA after immunizing rats, rabbits and mice with SF-RI 1. Repeated endotracheal administration of SF-RI 1 resulted in a humoral antibody response in three out of eight rabbits. After treatment of 34 preterm infants with SF-RI 1 (50-200 mg/kg), a humoral immune response to SF-RI 1 could not be detected. In-vitro restimulation of peripheral blood lymphocytes with SF-RI 1 after primary in-vivo administration did not result in cell proliferation as measured by 3H-thymidine incorporation. SF-RI 1 did not stimulate peripheral blood lymphocytes of neonates in vitro. The mitogenic response of these cells to stimulation with PHA, ConA or PWM was heavily impaired in the presence of SF-RI 1 concentrations increasing from 0.04 to 4 mg/ml. These data indicate that SF-RI 1 is immunogenic and that it may have an influence on lymphocyte proliferation in vivo.


Changes in cerebral oxygenation and cerebral blood volume during endotracheal suctioning in ventilated neonates.

The effect of endotracheal suctioning on cerebral haemodynamics was investigated in 29 newborn infants with a mean gestational age of 31 weeks (range 25-40 weeks). Prior to one of two suctioning procedures, the inspiratory fraction of oxygen was increased by 10%. Brain oxygenation and total haemoglobin concentration were estimated continuously by near infrared spectroscopy. Mean arterial blood pressure, arterial blood oxygen saturation and carbon dioxide tension were recorded simultaneously. Brain oxygenation decreased in parallel with arterial oxygen saturation during suctioning. Preoxygenation ameliorated the decrease in brain oxygenation and arterial oxygen saturation whereas there was no benefit with regard to the changes in total haemoglobin concentration, carbon dioxide tension or mean arterial pressure. Changes in total haemoglobin concentration were related closely to concomitant changes in carbon dioxide tension (p less than 0.0001) but unrelated to changes in mean arterial pressure or arterial oxygen saturation. Our findings suggest that cerebral blood volume may react to changes in carbon dioxide tension during endotracheal suctioning in mechanically ventilated neonates. Apparently, preoxygenation prior to suctioning does not ameliorate the stress in normoxic infants.


Low incidence rates by birth of symptomatic coeliac disease in a Danish population of children.

Using the original diagnostic criteria of the European Society for Paediatric Gastroenterology and Nutrition for symptomatic coeliac disease in children, we found an average incidence rate by birth of 0.10 per 1000 live births in a demographically homogeneous Danish population over a 15-year period. This incidence rate by birth is the lowest described in an epidemiological study in contrast with rates of 0.33 to 8 per 1000 live births found in other western countries. This difference may be explained by the different diagnostic criteria used for coeliac disease. Another contributing factor may be a late and gradual introduction of gluten-containing cereals into the diet of Danish infants.


Comparison of fiberendoscopy and Watson capsule for small intestinal biopsy in infants and children.

We analyzed the results of 607 small bowel biopsies performed over a seven-year period: 284 biopsies were obtained using a fiberendoscope and 323 biopsies using a Watson capsule. Three to six specimens were removed during endoscopy. The biopsy fragments obtained with the fiberendoscope were deeper than those obtained with the Watson capsule (p less than 0.0001) and were more often located in the duodenum (p less than 0.0001). The failure rate of the fiberendoscope biopsies (1%) was lower than for the Watson capsule biopsies (9%) (p less than 0.0001). Multiple biopsies increased the diagnostic value of fiberendoscopy which was 95% versus 85% for Watson capsule. The mean duration of the endoscopic procedure recorded in 30 children was 6.5 min for four to six samples, i.e. 1.5 min per biopsy specimen. Fiberendoscopy appears to be an efficient and safe method for performing small bowel biopsies in infants and children.


Normokalaemic pseudohypoaldosteronism is present in children with acute pyelonephritis.

The present study demonstrates that renal tubular unresponsiveness to aldosterone, without associated hyperkalaemia, is present in children with acute pyelonephritis. We studied 32 children with a diagnosis of acute pyelonephritis established by high fever, flank pain/tenderness, increased blood levels of C-reactive protein and significant Escherichia coli growth in the urine culture. Renal tubular function tests and determinations of plasma renin activity and aldosterone concentration were performed at diagnosis (study 1), after three days of iv gentamycin (study 2) and after 21 days of antibiotic therapy (study 3). Findings were compared to those present in 32 normal children of similar age. Despite normal plasma potassium concentration, fractional potassium excretion and transtubular potassium concentration gradient were significantly decreased in studies 1 and 2, becoming normal in study 3. Decreased renal potassium excretion coexisted with increased values for plasma renin activity and aldosterone concentration. In study 3 these hormones remained elevated only in patients with scarred kidneys. The functional alteration present in acute pyelonephritis may be directly caused by the interstitial inflammation or be mediated by some E. coli endotoxin.


Intrauterine growth in children with cerebral palsy.

The risk of cerebral palsy in connection with intrauterine growth retardation has been analysed in a case-control study. The case series comprised 519 children with cerebral palsy born in 1967-1982 in the west health-care region of Sweden and the control series 445 children born during the same years in the same region. The risk of cerebral palsy in small-for-gestational-age infants was significantly increased in term and moderately preterm infants. The highest proportion among infants with cerebral palsy born at term was found in tetraplegia, followed by diplegia and dyskinetic cerebral palsy. It was concluded that small for gestational age on the one hand reflects early prenatal brain damage, and on the other mediates prenatal risk factors compatible with foetal deprivation of supply and also potentiates adverse effects of birth asphyxia and neonatal hypoxia.


Danish primary schoolteachers' knowledge about asthma: results of a questionnaire.

We present an anonymous questionnaire inquiry involving 334 primary schoolteachers in the Randers area with the purpose of elucidating teachers' knowledge about asthma. To a series of statements about asthma, the teachers answered yes, no or don't know. A limited knowledge of different aspects of asthma in children was found, although 57% had asthma children in their classes. Specially limited was knowledge about medical treatment. Five percent had received proper instruction about asthma and had a significantly better knowledge of medical treatment (p less than 0.001-0.05). Only 57% knew that wheezing after physical exertion is a strong indicator of asthma and only 33% knew that exertion in cold weather increases the risk of an attack. It is recommended that instruction in children's diseases, especially asthma, is introduced in teacher training colleges.


Growing up in Uppsala. The role of public services in identification and treatment of health and adjustment problems. Part I. Definition and classification of dependent variables.

A comprehensive prospective longitudinal study of health, development and social adjustment from the age of four to 18 years has been carried out in Uppsala, Sweden. This report presents the accumulated psychosocial burden up to 18 years of age for all 1715 children born in 1965 and resident in Uppsala from age 10 to 18 years. The psychosocial burden up to 18 years of age was assessed through analysis of records and register information from school health services, authorities for care of the handicapped, the Department of Child Psychiatry, social agencies and legal authorities. The analysis showed that 11.8% of the adolescents had a severe psychosocial burden up to the age of 18 years which could hamper their future life as adults. It was possible to categorize the whole birth cohort into one subgroup without manifest psychosocial problems and five different subgroups with serious problems: the six groups (severe mental or physical handicap, antisocial behaviour, psychiatric problems, social support, multiple problems, "normal") had specific profiles concerning sex distribution, symptoms, social background, utilization of care services and delinquency.


Growing up in Uppsala. Part II. Could adolescents with severe psychosocial problems have been identified by symptoms observed in school at age 10 years?

The study comprised all 1715 children born in 1965 and resident in Uppsala at age 10 and 18 years. Data were collected through teacher interviews and analysis of school health records in grade 3 at the age of 10 years. The psychosocial burden up to the age of 18 years was assessed on the basis of all registered contacts with official institutions outside school (authorities for care of the handicapped, Department of Child Psychiatry, social agencies, legal authorities). Approximately 12% of the adolescents were clearly in a situation of manifest psychosocial risk on the threshold of adult life. These adolescents were assigned to five mutually exclusive problem groups comprising different sex distribution, symptoms and utilization of institutional care. The analysis of the relationship between data from grade 3 and the psychosocial burden up to 18 years of age showed that the information available to the school did not permit reasonably secure predictions of the child's psychosocial situation at the end of adolescence. Observations in school of pre-adolescent children cannot be used as a basis for risk-group strategies aiming at concentrating early treatment measures and resources to a restricted number of children at risk. However, the prognosis is apparently serious for a limited number of 10-year-olds with serious problems in school.


Greek immigrant children in southern Sweden in comparison with Greek and Swedish children. III. Energy and nutrient intake.

Greek immigrant children (GI) belonging to the second generation of immigrants in Sweden have been compared with Swedish children (S) and Greek children in Greece (G) regarding energy and nutrient intake. Twenty-four-hour recalls were obtained in the homes of the families. The mean energy intake was the same in all three groups and met the Swedish Nutrition Recommendations. The energy distribution for protein, carbohydrates and fat was also similar. The fat intake was far above the recommended level in all groups. The GI and the G group had a significantly higher mean daily intake of monounsaturated fatty acids than the S group (p less than 0.05 and p less than 0.001, respectively). Children aged four to eight years in the GI group had a significantly higher nutrient density of retinol, vitamin D, riboflavin, vitamin B6, calcium, iron, magnesium and sucrose compared to the G group, but compared to the S group they had a lower nutrient density of retinol, vitamin D, ascorbic acid, niacin, vitamin B12 and selen. The GI children consumed more milk than the G children but as much as the S children and they had started to use enriched low-fat milk in Sweden. In conclusion, the food quality in the GI group was better than in the G group and much the same as in the S group, and, with few exceptions, it met the Swedish Nutrition Recommendations.


Clinical and electrophysiological reports in a case of early onset myotonia congenita (Thomsen's disease) successfully treated with mexiletine.

A sporadic event of myotonia congenita in a infant admitted to the Paediatric Clinic for frequent crises of apnoea, cyanosis, vomiting and difficult feeding is reported. EMG analysis was consistent with the dominant variety of myotonia congenita. Mexiletine therapy showed excellent results in reducing myotonic activity. It is worthwhile stressing that early symptoms may go unnoticed or may be misinterpreted and that information on the genetic form of the disease can be obtained also from the EMG analysis through a repetitive stimulation test.


The primary empty-sella syndrome and diabetes insipidus in a child.

The empty-sella syndrome is uncommon in pediatric patients. Hypothalamic-pituitary dysfunction is common in these patients but involvement of the posterior pituitary gland is very rare. We report a seven-year-old girl with empty-sella syndrome who first developed arginine-vasopressin deficient diabetes insipidus and then anterior pituitary gland deficiency. The empty-sella syndrome should be included among the causes of arginine-vasopressin deficient diabetes insipidus in pediatric patients.


Role of human blood monocytes in up-regulation of lymphokine (interleukin-2)-activated killer cell activity with cisplatin and FK-565.

The role of cisplatin (CP) and FK-565 in monocyte-mediated up-regulation of lymphokine-activated killer (LAK) cell induction by interleukin-2 (IL-2) was examined. Treatment of peripheral-blood mononuclear cells (PBMC) with CP or FK-565 in the presence of IL-2 resulted in a significant increase in LAK activity against NK-resistant Raji cells, as assessed by the 4-hour 51Cr release assay, depending on the dose of CP, FK-565 and IL-2. Up-regulation of IL-2-induced LAK activity by CP/FK-565 was significantly higher in whole PBMC as compared to PBMC depleted of monocytes. Addition of different numbers of monocytes to cultures of lymphocytes plus IL-2 along with CP or FK-565 resulted in a significant up-regulation of LAK activity depending on the number of monocytes added. Pretreatment of monocytes for 2 h with CP or FK-565 before addition of lymphocytes plus IL-2 resulted in significant up-regulation of LAK activity as compared to untreated monocytes. Culture supernatant of CP- or FK-565-treated monocytes also significantly up-regulated IL-2-induced LAK activity of lymphocytes. The results of the present investigation suggest that up-regulation of LAK activity by CP and FK-565 in human PBMC is monocyte-mediated. Further, our data demonstrate that tumor necrosis factor and IL-1 play an important role in the up-regulation of LAK activity by monocytes that have been pretreated with FK-565 or CP. These results indicate that CP and FK-565 may be useful in modulating the immune response during treatment of neoplasia with IL-2 and LAK therapies.


Signal transduction in activated natural killer cells and natural killer cells inactivated with sensitive targets.

We have recently demonstrated that the activation of NK cells involves a protein-kinase-C (PKC)-dependent step as well as a phosphatidylinositol (PI)-linked signal transduction system. We have also shown that when NK cells are incubated with sensitive targets for up to 6 h they lose their lytic potential and require IL-2 to regain their lytic activity. PKC is involved in many cell processes such as the transduction of hormonal signals and the machinery of cellular secretion and is activated by diacylglycerol and by a number of phorbol esters, including phorbol myristic acid (PMA). To reinforce the role of PKC in NK-cell-mediated cytotoxicity, we first showed that NK-CMC is inhibited by two agents that inhibit PKC activation, staurosporine and sphingosine. Next, we showed that antibody-dependent cellular cytotoxicity of NK-resistant targets was PKC dependent and Ca2+ dependent. Further, we showed that PMA plus ionophore alone could induce NK killing of resistant targets and that this killing was PKC and Ca2+ dependent. Finally, we contrasted the role of PKC in these activated cells to the role of PKC in cells that have been inactivated with a sensitive target, K562. We showed that PKC is not required for the IL-2-dependent reactivation of NK cells but that Ca2+ is required. To further determine what event in signal transduction is inactivated by K562, we showed that inactivated NK cells do not turnover PI in response to K562 stimulation.


Epidemiology considerations in peptic ulcer disease.

Peptic ulcer disease is a common clinical problem. The lifetime risk of peptic ulcer disease is at least 10%. Millions of Americans are affected each year, imposing a major economic burden on the health care system. The overall hospitalization and mortality rates for peptic ulcer disease seem to have decreased substantially over the past few decades. There is much to suggest, however, that these changes were occurring even before effective medical therapy became available. In addition, other influences, such as increasing age of the population at risk, changes in smoking prevalence, and increasing use of nonsteroidal anti-inflammatory drugs, have impacted on the changing epidemiology of peptic ulcer disease. Hospitalization rates for patients with complications of peptic ulcer disease have remained relatively stable or, especially in the case of elderly women with gastric ulcers, increased significantly in recent years. Understanding the epidemiology of peptic ulcer disease will allow improved assessment of the effects of medical and surgical therapy and, hopefully, provide better clues to the etiology of this diverse group of diseases.


Population dynamics of natural killer cells and T lymphocytes in murine spleen and bone marrow during the development of erythroleukemia: the effect of indomethacin.

Quantitative and functional methods were used to assess changes in NK (ASGM-1+, nonblast, lymphoid) cells and cytotoxic/suppressor T (Lyt-2+) cells in the spleen and bone marrow (BM) of normal DBA/2 mice and/or mice bearing early (7 days)- and late (14 days)-stage erythroleukemia +/- concomitant oral administration of indomethacin. In normal mice, indomethacin increased ASGM-1+ cells in the spleen and BM by 7 days of continuous drug administration relative to control, while the numbers of Lyt-2+ cells in both organs remained unaltered. The presence of a tumor in untreated mice resulted, by 14 days development, in elevated numbers of ASGM-1+ cells in the spleen and BM. Lyt-2+ cells in the spleen increased transiently (7 days) in tumor-bearing (TB) mice and returned to normal levels by 14 days TB. No difference in either early- or late-stage TB was observed in the numbers of Lyt-2+ cells in the BM. In spleen and BM of TB mice, indomethacin increased ASGM-1+ cells by 7 days TB. ASGM-1+ cells returned to untreated levels by 14 days TB in both organs in spite of continued drug treatment. The numbers of Lyt-2+ cells in both the spleen and BM of 7 days indomethacin-treated TB mice were reduced relative to those of untreated TB mice. By 14 days TB, drug treatment had no effect on Lyt-2+ cell numbers in the spleen or BM. Spontaneous cytolytic activity of the spleen and BM did not always parallel changes in the numbers of ASGM-1+ cells, indicating the presence of late, immature, prelytic NK cells among the ASGM-1+ cell numbers. The results demonstrate that the numbers of NK and cytotoxic/suppressor T cells are markedly but differentially affected during tumor growth and/or time of exposure to indomethacin in an organ-specific manner.


Stress ulcer.

Stress ulceration is a recognized cause of morbidity and mortality in the critically ill patient. These superficial lesions occur in both the stomach and duodenum within hours after admission to a critical care unit. Although the pathogenesis of stress ulcer is not well understood, it is now established that a decrease in mucosal pH below 6.5 is essential for the induction of stress ulceration. This drop in mucosal pH can result from high luminal hydrogen ion (H+) concentration, increased permeability of the mucosa for H+, or defective neutralizing ability of the mucosa. Local and systemic factors that modulate mucosal blood flow are also important in the development of stress ulceration. It is estimated that 20% to 30% of patients with stress ulcer develop significant gastrointestinal bleeding that requires blood transfusion. The routine administration of H2-receptor antagonists has become standard practice in most intensive care units to prevent the development of stress ulcer and gastrointestinal bleeding, although in many cases their efficacy is unproven. Antacids are clearly effective and remain the gold standard with which alternative therapies are compared. The best treatment for stress ulceration is identification of the patient at risk and institution of prophylaxis. For patients who bleed from stress ulceration, conservative measures should include aggressive treatment of the underlying medical problem and neutralization of acid.


Effect of recombinant murine tumor necrosis factor on the generation of natural killer cells in bone marrow cultures.

We investigated the possible role of tumor necrosis factor-alpha (TNF-alpha) in the interleukin-2 (IL-2)-dependent generation of natural killer (NK) cells from bone marrow precursors. TNF-alpha synergistically augmented both cytotoxic activity against NK-sensitive targets and cell number at the end of the 7-day incubation period. After this time, NK activity was not induced by TNF-alpha in the absence of IL-2. The cytotoxic cells generated by IL-2 + TNF-alpha had the phenotype of mature NK cells, including expression of NK-1.1, asialo-GM1, Ly-5, LFA-1 and Thy-1. TNF-alpha was also able to up-regulate the mRNA expression for the IL-2 receptor alpha-chain (P55) as well as the mRNA expression of c-myc protooncogene. Blocking studies with monoclonal antibodies against the alpha-chain P55 of the IL-2 receptor confirmed the functional role ascribed to IL-2 in the in vitro generation of NK cells from bone marrow cultures. Additional proliferation studies demonstrated that the up-regulation of c-myc protooncogene was associated with an increased uptake of thymidine. These data indicate that the TNF-alpha-induced increase of IL-2-dependent NK cell generation from bone marrow precursors was associated with an augmented proliferation and an up-regulation of mRNA expression for IL-2 receptor and c-myc protooncogene.


[Clinical and experimental study of semen Persical decoction for purgation with addition in type II diabetes mellitus].

This paper reported the results of clinical observation on a treatment with Semen Persical decoction for purgation with addition (SPDPA) in type II diabetes mellitus. The effective rate of SPDPA on 106 cases of noninsulin dependent diabetes mellitus (NIDDM) was 79%. The efficiency of SPDPA was equivalent to glyburide. From the experimental study, it can be concluded that SPDPA could reduce blood sugar and relieve symptom in diabetic patients and rats. Its mechanism may be due to improving secretion of insulin, inhibiting production of glucagon, repairing insular endocrine cell, increasing endocrine pellet of insular B cell and improving composition of hepatic glycogen. In traditional Chinese medicine theory, the mechanism of therapeutic action of SPDPA in diabetes mellitus is based on synergistic regulation of benefiting Qi and nourishing Yin, activating blood circulation to dissipate blood stasis and loosening the bowel to relieve constipation.


[Determination of partially cellular and local immune function in patients with spleen deficiency syndrome].

The immune function status as reflected by the peripheral blood OKT system T cell subset classification and the lymphocyte in vitro interleukin 2 (IL2) secretory function were determined in 30 patients with Spleen deficiency syndrome and 20 normal subjects in this study. Experimental results decreased cellular immune function and disturbance of immune regulatory mechanism in the patients. Its manifestations were decreased number of total T lymphocytes and helper T cells (Th), relatively increased suppressor T cells (Ts), abnormal rate of Th to Ts, no marked change of IL2 secretory function of T cell in vitro, increased SIgA level before stimulating with acid but marked decreased SIgA level after stimulating. All these suggest the compensatory stage of local immune function.


[Effect of aging on the syndrome of essential debility and evil reality--an exploration on mechanism of senility by traditional Chinese medicine].

Effect of aging on essential debility and evil reality was explored by an epidemiological investigation of clinical syndrome with TCM in 878 cases of middle and old-aged patients, inquiring into their relation of senility with visceral weakness and stagnation of Qi, blood stasis, and phlegm turbid. The results indicated that (1) several viscera, feeble and damaged, were the basis of senility, and the feeble kidney was the stress; (2) the syndrome of stagnation of Qi, blood stasis and phlegm turbid speeded up process of senility. The mechanism of feeble phenomena appeared in the middle-old aged patients is that with the rise of age in the patients observed there was interaction between the feeble visceral function and the syndrome of stagnation of Qi, blood stasis and phlegm turbid, that is, there was interaction between essential debility and evil reality. Essential debility may lead to evil reality and the latter will worsen essential debility. Thus, on repeating themselves in alternate cycles, a systemic hypofunction will be formed up to exhaustion. Therefore, "to nourish essential debility first and to purge evil reality second" should be considered as an essential direction of preparing anti-aging drugs in TCM.


[Combined gu chi wan and spiramycin in the treatment of periodontal disease].

The purpose of the present study is to evaluate clinical effect of the integrated traditional Chinese and western medicine in the treatment of periodontal disease. 90 patients suffered from mild to advanced periodontitis were divided into two groups. The gu chi wan (tooth firming pills) group was administered tooth firming pills 4 mg twice daily for 3-6 months combined with spiramycin 0.2 four times daily for 5 days and routine periodontal treatment. The spiramycin group, as control, was administered spiramycin 0.2 four times daily for 5 days combined with routine periodontal treatment. The clinical parameters: GI, PLI, PDI and serial radiographs were checked on the beginning of study and through 3, 6, 12, 24 months follow up study. The results showed that the GI and PDI of the gu chi wan group decreased significantly than the control group (P less than 0.001). The inflammatory recurrence rate was 33% in the control group and 12% in gu chi wan group and the serial radiographs demonstrated that a higher incidence of bone fill occurred in gu chi wan group than in the control group (P less than 0.01).


[Influence of application-impulse stimulation treatment on airway's reactivity with asthma].

The authors observed the effect of application-impulse stimulation treatment on 184 patients with asthma, using the main index of airway's reactivity, in a randomized, double-blind, placebo-controlled parallel study. The airway's reactivity of the treated group got improved significantly (P less than 0.001), but that of the control group was not changed sharply (P greater than 0.05). The airway's reactivity and clinical effect between the groups have significant meaning (P less than 0.001, P less than 0.001). The result suggests that application-impulse stimulation treatment is a safe and effective method for preventing and treating asthma.


[Stimulative effect of electro-blunt-tip needle on the points of human skin and its analgesia in rats].

The stimulative effect of electro-blunt-tip needle (a kind of electroacupuncture stimulating on the surface of skin point) on the 120 points of human skin was observed in 12 healthy volunteers. It was shown that the substitute typical electro-blunt-tip needle stimulator and its special pen-like electrode by a general electroacupuncture stimulator and a tape-like adjustable electrode and similar therapeutic effects. It was found in animal experiment that the pain threshold determined with rat tail flick analgesia method was raised by electroacupuncture and electro-blunt-tip needle, the optimal stimulating time being 10-20 minutes for both, the percentages of the maximal changing pain threshold being 206.6% and 175.4%, and the corresponding voltage being 2v and 7v respectively.


[Flow cytometric analysis of the garlic oil effect on DNA content of cancer cell cycle].

Flow cytometry (FCM) is a new technique developed in the recent decade. This technique may measure DNA content of 5000 cells per second and trace the dynamic changes in cell proliferation cycle and offer a hint for designing clinical treatment protocol, monitor prognosis and elucidate the mechanisms of antitumor drugs. The authors previous studies showed significant effect of garlic oil on prolongation of life expectancy and inhibition of tumor growth in mice. Using FCM the authors analysed the effect of garlic oil on cell cycle in S180 tumor cells, 2-6 hrs after single administration or multiple administration the cell number in S phase rapidly decreased, in G1 phase increased. This suggests garlic oil may blockade cells to progress from G1 phase to S phase and result in accumulation of cells in G1 phase and directly inhibit the synthesis of DNA and the cell cycle. Theoretical basis for clinical application was offered and some aspects of antitumor mechanism of garlic oil were elucidated.


[Pharmacologic studies of antimotion sickness actions of ginger].

The pharmacologic actions related to antimotion sickness effects of ginger (Zingiber officinale Roscoe.) were studied. There was no significant effect on parameters of rotatory movement-induced electronystagmogram of rabbit after intravenous (i.v.) infection of ginger juice. The low amplitude fast wave pattern of electrocorticogram of rabbit changed to high amplitude slow wave pattern after i.v. injection of ginger juice. Rabbit gastric contraction in situ was shortly suppressed after ginger juice i.v. administration. In the isolated rat fundus strip preparations, however, ginger juice reduced the spontaneous contractile frequency, and enhanced the spontaneous contractile amplitude, which was followed by inhibition. Ginger juice produced longitudinal contraction of the guinea-pig isolated ileum, which was followed by rapid tachyphylaxis. This contraction effect was not affected by hexamethonium and 5-HT, but could be inhibited by cold storage, hyoscine, morphine, diphenhydramine, promethazine and substance P desensitization. Naloxone could eliminate this inhibition produced by morphine. By using dose-response relationship plot, non-competitive antagonisms were observed between ginger juice and Ach and between ginger juice and histamine in isolated guinea-pig ileum. It is suggested that the pungent constituents of ginger release substance P from sensory fibres. The released substance P in turn either stimulates cholinergic and histaminic neurons to release Ach and histamine, respectively, or produces direct muscle contraction by activating M and H1 receptors correspondingly. It is proposed that after being excited by substance P, M and H1 receptors are inactive temporarily and unable to be excited by agonists, therefore, ginger juice exhibits anticholinergic and antihistaminic action. Ginger juice produces antimotion sickness action possibly by central and peripheral anticholinergic and antihistaminic effects.


[Treatment of severe chronic hepatitis B by combination of traditional Chinese medicine and Western medicine therapy--with an analysis of 122 cases].

This paper is a summary report of treatment of 122 cases of severe chronic hepatitis by combination of TCM and WM therapy. Among them, 44 cases were in their early stage of disease, 66 in their intermediate stage, and 12 in their advanced stage. They were treated by basic therapy with hepatic cell growth factor (HGF) added to some of them. The patients were divided into two groups, one receiving treatment of blood-cooling and detoxifying Chinese herbs, the other receiving treatment of heat-clearing and detoxifying Chinese herbs. The treatment was effective for 52 cases, and ineffective in 70 cases. Values of serum bilirubin (SB) and prothrombin activity (PA) and the time of initial consultation had marked influence on their prognoses. No difference was found in total mortality between the two groups, also no difference was found in mortality between the former and latter group with or without the addition of HGF at the early stage of disease. The survival rate of the former group was significantly higher than that of the latter one with HGF added at their intermediate stage of disease, which were 60% and 14.7% respectively, P less than 0.01. The conclusion is that as diagnosis and treatment were based on the Syndrome Differentiation of patients at intermediate stage of disease, Chinese herbs will be effective in many ways such as lowering and eliminating jaundice, improving symptoms and signs, preventing and curing complications, curing coexisting diseases, and repairing pathological lesions.


[Modulating effects of extractum semen Persicae and cultivated Cordyceps hyphae on immuno-dysfunction of inpatients with posthepatitic cirrhosis].

The treatment of 65 cases (with 20 cases in the control group) of patients with post-hepatic cirrhosis by Extractum Semen Persicae (ESP) and cultivated Cordyceps Hyphae (CH) was reported, and the adjustment of the disorders of both cellular and humoral immune functions was investigated. The results indicated apparently that the restrained cellular immune function, among which the rate of lymphocyte transformation, the NK cell's function, CD8+ and CD4+ cells in the patients' group were lower than those in the healthy group. The CD8+ cell count was positively correlated with lymphocyte transformation rate and the humoral immune hyperfunction revealing that the levels of IgG, IgA, ssIgA and CIC of the patients were abnormally higher than those of healthy subjects, while level of C3 was negatively correlated with that of CIC. After the treatment, rate of transformation, function of NK cell, CD8+, CD4+ cells and the ratio of CD4+/CD8+ improved; levels of IgG, IgA, ssIgA and CIC reduced, while those of C4 and C3 rose. These results revealed the ESP and cultivated CH could modulate the cellular immune function, inhibit the humoral immune hyperfunction and increase serum complement level in patients with post-hepatitic cirrhosis, which were correlated to the improvement of hepatic function. Disorder of immune function was one of the important causes of liver cell necrosis, inflammatory cell infiltration and fibroplastic proliferation. The results showed that the drugs are of some value in the treatment of patients with post-hepatitic cirrhosis.


[Characteristics of clinical pathology and changes of liver function in blood stasis syndrome in liver diseases].

The experimental study on 30 patients of the Blood Stasis Syndrome of liver diseases was discussed in this paper. Two characteristics were found. One was the pathological feature which manifested as follows: (1) DIAGNOSIS: the Blood Stasis Syndrome of liver disease was mainly diagnosed in the chronic active hepatitis and the early stage of cirrhosis of liver, while that of non-Blood Stasis was mainly observed in the chronic persistent hepatitis (P less than 0.01); (2) Pathological change: The histological changes such as piecemeal necrosis, bridging necrosis, the destruction of limiting plate, eosinophilic change, etc. It was more obvious in the Blood Stasis group than that with non-Blood Stasis Syndrome (P less than 0.01), (3) The manifestation of Blood Stasis Syndrome was not in parallel with the severity of the liver disease. The another characteristic was the changes of liver function, which expressed more markedly in the Blood Stasis group with higher level of SGPT, lower ratio of A/G and increased globulin, they were more obvious than that in non-Blood Stasis group (P less than 0.01).


[Diagnosis on blood stasis in viral hepatitis by computer multi-criteria analysis--an analysis of 63 cases].

This study is based on the criteria for diagnosis of Blood Stasis formulated by the international conference held in Beijing in 1988. The patient that has one, two, three, and over four items could be diagnosed as I-, II-, III-, IV-degree hepatic Blood Stasis respectively. The data of pathologic findings from liver biopsy, nail-fold microcirculation, blood rheology, hepatogram, B-ultrasonography and blood biochemical examination were stored in computer to carry on single and multiple factor analysis in order to evaluate these indexes in the diagnostic value of hepatic Blood Stasis. Stepwise regression analysis formula was used to determine the severity of Blood Stasis. By substitute related data into the formula, mathematical model of hepatic Blood Stasis was compared with clinical Syndrome Differentiation, it revealed that 59 cases were coincided with each other (93.6%). According to numerical value, the increase and decrease of the pre-and posttherapy, the curative effect of promoting blood circulation to remove Blood Stasis could be determined to provide some references for diagnosis and treatment of hepatic Blood Stasis and prevention and treatment of hepatic fibrosis.


[Ischemic apoplexy treated with acupuncture using the principle of replenishing qi and promoting blood circulation].

32 cases of ischemic apoplexy were treated by acupuncture using the principle of replenishing Qi and promoting blood circulation. The clinical efficacy and the change of nail-fold microcirculation and hemorheology before and after the treatment were observed. The total effective was 93.75%. Before the treatment, the microcirculation of nail-fold was markedly abnormal, and the criteria of hemorheology was abnormally elevated. But after the treatment, together with the recovery of nail-fold microcirculation and hemorheology, the clinical symptom and sign of the patients also improved. Thus it showed that acupuncture method had the function of changing the microcirculation and the hypercoagulability of the patients' blood, further promote the recovery of the function of affected cerebral tissue.


[Determination of blood hormone levels in patients with "spleen deficiency" syndrome after major operations].

The character of "Spleen Deficiency" (SD) Syndrome was evaluated by determination of the blood levels of various gastroenteropancreatic hormones and thyroid hormone. Patients of SD group were diagnosed clinically by Syndrome Differentiation, and those of the normal group were volunteer blood donors. RIA was used to determine all the hormones except T3RUR. TT3, FT3 of the SD group were very significantly lower than that of normal group (P less than 0.001). The levels of rT3 in SD group were significantly higher than normal control (P less than 0.05). For TT4, T3RUR and FT4, there were no significant differences between the two groups. Among the gastroenteropancreatic hormones, gastrin in the SD group was significantly lower than that of the normal group (P less than 0.01), motilin was very significantly higher (P less than 0.002). The value of pp was similar in the two groups, under physiological conditions, however, in adults the hormone tended to increase with age. Since the average age of patients in the SD group was 54.9, and that of the normal group was 38.5, remarkably lower values were seen in the SD group. Other hormones such as VIP, SST, glucagon and insulin were of no significant difference between the two groups. The result showed that thyroid hormone and some of gastroenteric hormones in the blood of patients with SD after major operations were quite different from normal, and was common to patients with general SD Syndrome, with low T3 syndrome.(ABSTRACT TRUNCATED AT 250 WORDS)


[Antagonistic effect of re du qing on damage of hepatocytic microsomes in rabbits with endotoxin-induced disseminated intravascular coagulation].

In this study, the generalized Shwartzman reaction of rabbit induced by endotoxin of Escherichia Coli was built as DIC models. The experiment showed that the levels of lipid peroxide (LPO) in the hepatocytic microsomes of model group were increased significantly, whereas the cytochrome P-450 (Cyto. P-450) contents and aniline hydroxylase activities were obviously decreased. In the Re Du Qing (RDQ) group and dexamethasone group the decrease of LPO in hepatocytic microsomes as well as the reduction of Cyto. P-450 contents and aniline hydroxylase activities were alleviated. Furthermore, the correlation analysis indicated a significant negative correlation between levels of LPO in microsomes and the Cyto. P-450 contents as well as aniline hydroxylase activities. This study indicates the LPO may play an important role in the damage of hepatocytic microsomes and RDQ could prevent hepatocytic microsomes from injury of rabbits with endotoxin-induced DIC.


[Experimental study on yiqi-huoxue therapy of liver fibrosis].

The experimental hepatic fibrosis was treated with YiQi (reinforcing Qi, YQ) Huoxue (activating blood circulation, HX) principle which was consisted of astragalus membranaceus, Ligusticum wallichii, paeonia lactiflora, etc. After stimulation with CCl4 over four months, the Wistar rat developed liver fibrosis. Rats were divided into the normal control, the toxifying control, YQHX group and HX group. The experimental period lasted over four months. (1) Mortality of animal: Both toxifying control and HX group reached 50%, while YQHX group was 16% only. These results suggest that YQHX agents could strengthen the body resistance; (2) The determination of serum SGPT: The mean levels in toxifying control were 39.3 +/- 39, in HX group 43.7 +/- 12.9, while in YQHX group 29.0 +/- 7.6 (units/Lserum). These results indicated that YQHX agents had the function of protecting the liver and lowering the activity of SGPT (P less than 0.01); (3) Measuring the contents of hepatic collages: The mean levels in toxifying control was 38.9 +/- 3.3 (mg/g liver), while in HX group and YQHX group 28.7 +/- 2.2 and 22.7 +/- 1.1 (mg/g liver) respectively. The results indicated that the YQHX agents had the best results in treating hepatic fibrosis; (4) Observation with hepatic histopathology: The degree of hepatocyte degeneration and necrosis in YQHX group was milder than that in toxifying and HX group. These observations revealed that YQHX agents possessed the function of protecting liver. There was severe liver fibrosis in toxifying control, but the degree of liver fibrosis in YQHX group was significantly milder than that in toxifying control.(ABSTRACT TRUNCATED AT 250 WORDS)


[Perioperative hemostatic problems in children].

During childhood preoperative coagulation diagnosis is performed to prove or to rule out an inborn coagulation disorder or an acquired v. Willebrand disease. The coagulation system of the newborns differs considerably from that of the adults as well as the time in which the single parameters reach adult values. Reducing the coagulation screening to the determination of aPTT and Quicktest neglects severe hemostaseological disorders easily, e.g. v. Willebrand disease which often in childhood is to be observed and often combined with normal aPTT values. Probably sometimes children affected with hypertrophy of the adenoids have temporary aPTT prolongations combined with normal values for the other coagulation parameters possibly due to lupus inhibitors. When children with deficiencies of coagulation factors need high doses of coagulation concentrates the number of laboratory controls may be reduced by determination on recovery and half-life period some times before.


Blood component processing technique and plasma quality.

To maintain a closed system during the preparation of blood components, including the removal of buffy coat, many centers use a quadruple blood bag additive solution system which in this study has been reduced to a cheaper triple bag system. The buffy coat and plasma were after centrifugation transferred to the first satellite bag and, after a second spin, the plasma separated from the buffy coat was transferred to the second satellite bag and stored for a fortnight at 4 degrees C. This resulted in a statistically significant increase in platelet factor 4 and elastase activity levels. No significant changes were found in the levels of C1-esterase inhibitor and kallikrein inhibiting activity, thrombin-antithrombin complexes, soluble fibrin, fibrinopeptide A and spontaneous proteolytic activity. The changes observed must be regarded as clinically insignificant. The platelet count is low enough to meet the requirements for platelet poor plasma. Using this blood component separation technique, one can reduce the CPD/additive solution 4-pack blood bag system to a less expensive 3-pack blood bag system.


Biocompatibility of synthetic oxygen carriers and fluorosurfactants.

A system for testing the biocompatibility of synthetic oxygen carriers is described including tests of hemolysis, complement activation and proliferation of cell lines or lymphocytes. Of 17 surfactants tested in this system, 6 were compatible in all tests while the other compounds showed individually differing patterns of incompatibility. We conclude that, in order to conduct a meaningful screening, a series of different assays has to be applied. In addition to our system, further assays, covering cytokine induction and phagocytosis should be attempted.


[The elimination of carboxyhemoglobin--gender-specific and circadian effects].

We investigated sex-specific and circadian variations of carboxyhemoglobin (COHb) elimination in healthy volunteers. COHb elimination follows an exponential function with a baseline found in nonsmoking women at 1.72% and in nonsmoking men at 1.79% COHb saturation. During the day, the elimination half-life time (t1/2), determined by eleven hourly samples after stopping smoking was significantly (Wilcoxon-test, p less than .01) shorter in women (3.2 +/- .4 hours, mean +/- standard deviation, n = 7) than in men (4.5 +/- 0.4, n = 6). During the night, too, t1/2 calculated from 2-point measurements after the smoking halt was significantly (Wilcoxon-test, p less than .05) shorter in women (4.3 +/- 1.1, n = 4) than in men (8.0 +/- 1.6, n = 5). In men, t1/2 during the night was significantly longer (p less than .01, Wilcoxon-test) than during the day. The day/night difference may be caused by reduced alveolar ventilation during sleep. Less muscle mass and, therefore, less myoglobin as a depot for carbon monoxide may account for the shorter half-life time of carboxyhemoglobin in women. We conclude that these variations of COHb-elimination should be taken into account when a smoking halt is advised, for instance, preoperatively.


Selection of unrelated bone marrow donors: does the current procedure warrant complete MHC class II identity?

Bone marrow transplantation from unrelated donors is being used increasingly for the treatment of patients with leukemia and several other hematologic disorders. Selection of unrelated bone marrow donors currently relies on serological HLA identity and negative mixed lymphocyte reactions between donor/recipient pairs. As serological HLA-DP typing is not feasible, we used the HLA-DPB1 oligonucleotide typing method to investigate whether the current selection procedure can guarantee complete MHC class II identity. In 40 consecutive patients, one third (62/193) serologically HLA-A, -B, -C, -DR and -DQ identical donors were found to be MLC negative with a relative response below 5%. HLA-DPB1 oligonucleotide typing of these MLC negative donors revealed that again only one third (20/62) was also identical for DP with their presumptive recipients. In the majority of pairs a disparity in graft-versus-host direction or in host-versus-graft direction of at least one allele was seen. These data indicate that in spite of the strict MLC criteria used, the current procedure did not warrant complete MHC class II identity. This implies that oligotyping for DPB1 can improve matching and should be introduced for typing of volunteers. We speculate that DP differences may contribute to the higher incidence of graft-versus-host disease or graft rejection in unrelated transplants.


Automated measurement of cell destruction: the Patimed system in comparison with conventional HLA typing and the cell mediated lympholysis test (CML).

The key role of HLA polymorphism in the pathogenesis of many diseases and in determining compatibility in transplantation makes it desirable to employ automated systems for identifying HLA polymorphism. For more than one year we have used the photometer system Patimed commercially available from the Leica company. We compared the results of the conventional cell-mediated lympholysis test and conventional serological HLA typing with the results obtained by the Patimed system. Despite some good preliminary results it would be premature, in our opinion, to recommend the Patimed system for routine assessment of killer cell activity whereas we can definitely recommend the Patimed system to define HLA class I and class II molecules serologically for research or routine purposes.


[Gene amplification with PCR and sequence-specific HLA oligotyping].

Genetic polymorphism in the HLA-class II region has been identified by the analysis of the polymerase-chain reaction (PCR) products using sequence-specific oligonucleotide (SSO). The PCR-SSO method permits precise and direct analysis of allelic variations with as little as 1 microgram of genomic DNA. The standardized, uniform hybridization and critical wash protocol of the Eurotransplant-typing kit [2] enables HLA-typing independent of gene expression and lymphocytes quality. One of the advantages of this technique is that the definition of subtypes is much better than is typing by serology.


One year's experiences with two different image analysis systems for automated reading of the contrast fluorescence test.

We have tested two different personal-computer based color image analysis systems for automated reading of the microlymphocytotoxicity test (LCT) for HLA-A,B,C-typing and screening. Over 17,000 single LCT-reactions were prepared using the simultaneous double fluorescent variant of the LCT (contrast fluorescence test, CFT). All tests were read visually by experienced laboratory staff members. For image analysis, an automated scanning system was used. In a first step, reactions were recorded on a videotape recorder using a color(CCD)-video camera. In a second step, the recorded reactions were analyzed with the two different image analysis systems by specifically developed programs. Good correlation (r = 0.89) of the score values assigned by digital image analysis with the visual tray reading was obtained. Since also the other main performance characteristics of the prototype system (throughput, reliability, compatibility) were acceptable for routine application, we may conclude that digital image analysis is a feasible and very interesting new technique for automated evaluation of the LCT.


[Changes in thrombocytapheresis concentrations caused by leukocyte depletion with polyester filters].

We studied platelet loss, leukocyte depletion, T/B-cell-ratio, platelet morphology and platelet function (aggregation, hypotonic shock reaction, retention and retraction) prior to and after filtration through two different polyester filters (Sepacell PL-5A, Pall PL100) as well as the posttransfusional platelet increment 1 h after transfusion in 8 pairs of single donor platelet concentrates (Cobe Spectra, Fresenius AS 104). Leukocyte depletion was effective (greater than 99.9%) and platelet loss acceptable in both filters, all tests showing no indication of a diminished platelet function by filtration.


Effective derosetting of mononuclear cells isolated by immunomagnetic beads.

A rapid and effective method for the derosetting of peripheral mononuclear cells (PMNC) isolated by CD8 monoclonal antibody (MoAb) coated immunomagnetic beads (IMB) is described. PMNC are positively selected and enriched by CD8-IMB and then derosetted from the IMB by a polyclonal antibody (PoAb) preparation against mouse Fab. Our experiments confirm the feasibility of this method with a high derosetting efficiency.


Filtration of buffy coat free red cell concentrates in additive solution.

Leukocyte poor RCC's (LP-RCC) are indicated in chronically transfused patients in order to prevent non-hemolytic transfusion reactions and HLA alloimmunization. In this study buffy coat free red cell concentrates (BCF-RCC) in additive solution (SAG-M) stored for four weeks were leukocyte depleted by filtration with three different filter systems (Erypur Optima (E), Sepacell R500 B (S) and, Pall RC, 50 TM (P)). The BCF-RCC's were prepared using 'bottom and top (BAT)' systems and automatic separation containing about 20% leukocytes and 5% platelets of fresh whole blood. The leukocyte concentration could be reduced to less than 5 x 10(6) per RCC with all filter systems equally: Leukocytes/RCC's: E .58 +/- .94, S .36 +/- .55, P .55 +/- .69 x 10(6). The leukocyte depletion was even in case of filtering two RCC's through one filter (double filtration) efficient enough in order to keep leukocyte contamination below the 'critical immunogenic load for leukocytes (CILL)'. But significant differences concerning the damage of red cells (free hemoglobin, LDH, HBDH) were measured which were even considerable: free hemoglobin E = 3.69 +/- 2.28, S = 1.31 +/- 1.24, P = 3.58 +/- 2.34 g/l. Double filtration was only performed with filter system S showing the best blood compatibility. But the second BCF-RCC also showed considerable hemolysis. Therefore, double filtration of RCC's only seems to be indicated under optimal conditions with blood compatible filters for selected patients. Bed side filtration cannot be recommended because of the risk of hemolysis that makes quality control necessary.(ABSTRACT TRUNCATED AT 250 WORDS)


The effect of posture and environmental temperature on cardiovascular reflexes in normal subjects and diabetes mellitus.

This study was undertaken to determine the effects of environmental temperature (17 degrees C or 27 degrees C) and posture on heart rate variability during forced sinus arrhythmia (deep breathing), and of environmental temperature on the heart rate and blood pressure responses to standing, and to the cold pressor test. A group of non-diabetic young subjects (23 +/- 5 years) and a group of older diabetic patients (54 +/- 15 years) were studied. At 27 degrees C, supine resting heart rate and systolic blood pressure were lower than at 17 degrees C in both diabetic and non-diabetic subjects, and heart rate variability was lower than at 17 degrees C in the non-diabetic subjects (p less than 0.01) with seven of 17 abnormal or borderline results (all seven subjects had normal values at 17 degrees C). Room temperature had no effect on supine heart rate variability in the diabetic patients. At 27 degrees C heart rate variability in the upright posture was significantly greater than supine (p less than 0.001) in the non-diabetic subjects. In the diabetic subjects, heart rate variability when standing at 17 degrees C increased such that five patients classified as abnormal when supine had normal values when upright. A similar pattern was observed at 27 degrees C, although only those with normal heart rate variability showed an increase on standing. Room temperature had no effect on the responses to the cold pressor test, or the blood pressure responses to standing, although the heart rate response was greater in both groups at 27 degrees C. This study shows that care should be taken to standardize the conditions under which heart rate variability is determined.


Parkinson's disease and weight loss: a study with anthropometric and nutritional assessment.

In order to investigate a putative weight loss in patients with Parkinson's disease, an anthropometric and biochemical study was undertaken. We compared body weight and indexes of fat [body mass index (BMI), tricipital skinfold] and lean [midarm muscle area (MMA), calf circumference] mass in men and women suffering from idiopathic Parkinson's disease with normal controls. We found that women suffering from Parkinson's disease exhibited a significant weight loss (-8.5%) and decreased calf circumference when compared with controls. A decrease (-4.3%) in total body weight was also found in men with Parkinson's disease but the difference did not reach the level of significance. Protein biochemical markers of nutritional status (albumin, prealbumin, retinol binding protein, transferrin) were normal in Parkinson's disease patients. The present study demonstrates the occurrence of weight loss in a large population of patients with Parkinson's disease. The putative mechanisms involved in the weight loss are discussed.