Use of selected visceral protein measurements in the comparison of branched-chain amino acids with standard amino acids in parenteral nutrition support of injured patients.

Twenty injured patients in the intensive care unit were randomized to receive parenteral nutrition with either 21% (STD) or 46% (HBC) branched-chain amino acids to compare the response of nitrogen balance (NB), somatomedin-C/insulin-like growth factor I (SMC), circulating fibronectin (FBN), and prealbumin (PA). NB was measured and serum collected for SMC, FBN, and PA on days 1, 4, 7, 14, and 21 of nutritional intervention. The treatment groups did not differ significantly for age, weight, injury severity score, trauma score, Apache II score, acute-phase protein concentrations, or type of injury. Comparison of baseline measurements revealed no significant differences in SMC, FBN, or PA. Both groups received similar doses of nonprotein energy and nitrogen. Baseline urea nitrogen excretion was slightly higher in the STD group (216 +/- 55 vs 268 +/- 54 mg/kg/day p = 0.049). Although NB was significantly improved over baseline during subsequent study days, there were no differences between groups after the day-1 measurement. SMC increased significantly from baseline on day 4 in the STD group, on day 7 in the HBC group, and on days 14 and 21 in both groups. There was no significant difference in SMC concentrations between groups on any day. Each group demonstrated a significant increase in PA from baseline on days 7, 14, and 21; however, no difference was seen when groups were compared. FBN increased significantly from baseline on day 14 in the HBC group and on days 7 and 14 in the STD group. FBN measurements were significantly different between groups on day 14 (STD, 179 +/- 71 vs HBC, 229 +/- 59 micrograms/ml; p less than 0.05). NB, PA, SMC, and FBN improve significantly during parenteral nutrition of traumatized patients. With the measured variables, there appears to be no significant difference between STD or HBC amino acids when used as part of parenteral nutrition in injured patients.

Effects of exogenous somatotropin on the concentration of minerals in the tissues of growing ram lambs.

Ten Suffolk ram lambs (mean BW 14.4 +/- 1.0 kg, 9 wk of age) were used to evaluate the effects of somatotropin (ST) on the concentration of minerals in tissues. Lambs were treated with daily injections of sterile saline (SAL; n = 5) or with bovine pituitary-extracted ST (.1 mg.kg BW-1.d-1; n = 5) for 13 wk. At slaughter at 22 wk of age, the liver, heart, kidneys, brain, spleen, lungs and testes were removed. Tissues were lyophilized, ground and analyzed for minerals. Daily treatment with ST had no influence on Ca, Na, K or the ratio of Na to K in tissues. Although P was lower (P less than .10) in the liver of ST lambs, the concentration of P remained within a normal range for sheep. The concentrations of both Mg (P less than .10) and Fe (P less than .05) were lower in the spleen of ST lambs. Splenic hypertrophy appeared to occur (P = .13) in ST lambs; the total splenic pool of Mg and of Fe did not differ (P greater than .10) between SAL and ST lambs. The concentration of several other minerals were lower in tissues of ST lambs: Cu in kidneys (P less than .10) and liver (P = .12); Zn (P less than .05) in liver, kidneys and lungs; and Mn in liver (P less than .05). By causing a reduction in the concentration of minerals in tissues, ST may increase the possibility of a metabolic insufficiency of a mineral. Exogenous treatment of animals with ST may modify the metabolic requirements for minerals and thereby increase dietary requirements.

[Detection of toxins of Clostridium perfringens type A in infected animals by using the ELISA test]. / Wykrywanie toksyn Clostridium perfringens typu A u zakazonych zwierzat przy uzyciu testu ELISA.

The aim of this study was to evaluate the usefulness of ELISA in toxin detection in guinea pigs experimentally infected with toxinogenic strain of Clostridium perfringens type A. The toxin was detected in blood serum and muscles from 12 hours after infection. The results obtained indicate the advantage of ELISA over to date methods used as immunofluorescence or microscopic examination of muscle exudate or sections. ELISA due to its high sensitivity rapidity and specificity allows to detect toxin in guinea pigs before clinical symptoms of gas gangrene are developed.

Fractionation and purification of some muscular and visceral ciguatoxins extracted from Carribean fish.

1. Cigautoxins (CTX) were extracted from flesh and viscera of seven large roving predatory fishes: Caranx bartholomaei, Caranx latus, Seriola dumerili, Alectis crinitus, Scomberomorus cavalla, Sphyraena barracuda and Gymnothorax funebris. 2. Generally each extract consisted of close-related CTX which were separated according to their polarity by Florisil column chromatography into a fast-acting CTX containing group and a slow-acting CTX containing group. 3. The shortest survival time of mice (ts) was low for the former group (less than 10 min) and high for the latter (greater than or equal to 29 min). 4. The level of purity had no influence on the range of ts values. The presence of these two CTX groups in different extracts did not from experimental conditions. 5. Attempts to convert fast-acting CTX to slow-acting CTX and vice-versa were negative. G. funebris and S. barracuda had an especially high content of unstable fast-acting CTX. 6. Purification of the slow-acting CTX was achieved by fast elution chromatography and Sephadex LH20 gel filtration. 7. The ts values of these CTX were identical for five species (40-44 min) but not for S. barracuda (29-32 min). 8. Thus ciguatoxic extracts from Caribbean fish were composed of several close-related CTX.

Transcription of the dystrophin gene in human muscle and non-muscle tissue.

The gene that is defective in patients with Duchenne and Becker muscular dystrophy consists of about 60 short exons scattered along a gigantic DNA region that spans some 2 megabase pairs. The encoded protein, dystrophin, was recently characterized as a component of muscle intracellular membranes of low abundance. The dystrophin messenger RNA is difficult to study in both normal and pathological tissue specimens because it is large (14 kilobases) and scarce (0.01-0.001% of total muscle mRNA). We report here that efficient in vitro co-amplifications of the mRNAs of the dystrophin gene and of a reporter gene, aldolase A, by the polymerase chain reaction procedure enables us to obtain a quantitative estimate of the dystrophin gene transcript. A processed, transcribed segment was thus detected in 13 different human tissues. It ranged from 0.02-0.12% of total mRNA in skeletal muscle to 25,000 times less in lymphoblastoid cells.

Immunohistochemical studies of cholecystokininlike peptides and their relation to 5-HT, CGRP, and bombesin immunoreactivities in the brainstem and spinal cord of lampreys.

The distribution of cholecystokinin (CCK)-like immunoreactivity in the brainstem and spinal cord of lampreys was studied by using CCK antisera with different properties. In the spinal cord, three separate systems reacted with CCK antisera: (1) A ventral and lateral fiber system descending from a group of neurons in the posterior reticular nucleus of the rhombencephalon was labeled by both a C-terminal-directed CCK antiserum and a monoclonal CCK antibody. (2) A dorsal root-dorsal column system of fibers originating from cell bodies in the dorsal root ganglia was labeled only by the C-terminal CCK antiserum. This CCK immunoreactivity could be abolished by preabsorption with calcitonin-gene-related peptide (CGRP), suggesting that it was due to cross-reactivity with a CGRP-like peptide. This system also contained 5-hydroxytryptamine (5-HT)-, bombesin-, and CGRP-like immunoreactivities. (3) An intraspinal system of 5-HT neurons was labeled with an antiserum to the midportion of CCK-33 but not by the other CCK antisera. The CCK labeling of this system was difficult to reduce by preabsorption with CCK peptide and thus appeared to be nonspecific. Groups of cell bodies in the middle reticular nucleus of the rhombencephalon, the reticular nucleus of the mesencephalon, and the hypothalamus were labeled by both the C-terminal and the monoclonal CCK antisera. The gut contained two types of CCK-like immunoreactivity, one of which appeared to be due to cross-reactivity with CGRP. A biochemical analysis showed that the content of CCK was low in the spinal cord compared to the brain, and these results agreed with the immunohistochemical findings.

Seasonal frequency of ciguatoxic barracuda in southwest Puerto Rico.

Ciguatoxicity of barracuda (Sphyraena barracuda) head, viscera and flesh tissues has been determined in 219 specimens caught along the southwest coast of Puerto Rico from March 1985 through May 1987. Twenty-nine percent of these specimens were toxic. Monthly frequencies of ciguatoxic barracuda showed an apparent seasonal variability, with peak values (60-70% toxic fish) in the late winter-early spring (January-May) and fall (August-November). Minimal frequencies (0-10% toxic fish) were observed during June-July and December. The most frequently toxic tissues in poisonous animals were the viscera and head. Viscera tissue was the only toxic tissue found in 31% of the poisonous fish assayed, and this tissue was poisonous in all toxic fish. In no case was a poisonous specimen found to have toxic flesh alone. Marked temporal variation in frequency of ciguatoxicity suggests that ciguatera toxins, at least in their active form, are not accumulated in barracuda tissues for extended periods of time. Variability in barracuda ciguatoxicity may reflect fluctuations in the toxicity of smaller reef fish prey, seasonal fluctuations in toxic benthic dinoflagellates and/or changes in the ability of the barracuda to detoxify ingested poisons or their precursors.

Immunohistochemical localization of insulin-like growth factor I in the adult rat.

Rabbit antisera against native human insulin-like growth factor I (IGF-I; somatomedin C) or a synthetic tetradecapeptide, representing the carboxyterminal amino acids 57-70 of human IGF-I, were used to map immunohistochemically the distribution of IGF-I immunoreactive material in adult rats. Both antisera were specific for IGF-I, as characterized by immunoabsorption, immunoblotting and radioimmunoassay. There was no cross-reactivity to IGF-II, relaxin or pro-insulin; substances having a high degree of structural homology with IGF-I. High IGF-I immunoreactivity was observed in spermatocytes of the testis; in oocytes, granulosa and theca interna cells of the ovary during early stages of follicle development; in some lymphocytes and in reticular cells of lymphoid and hematopoietic organs; in salivary gland duct cells; in the adrenal medulla, the parathyroid gland and the Langerhans' islets. Chondrocytes in the epiphyseal and rib growth plates and at articular surfaces showed strong IGF-I immunoreactivity. Brown but not white fat cells were stained. Nerve cells in the peripheral and autonomic nervous system showed faint to intense IGF-I immunoreactivity. In contrast, neurons and neuroglial cells in the central nervous system were generally negative; motor neurons being an exception. Erythropoietic, thrombocytopoietic and myeloic cells in the bone marrow showed IGF-I immunoreactivity, but only at defined developmental stages. Hepatocytes showed faint IGF-I immunoreactivity, but became more intensely stained after pretreatment with colchicine. The present results suggest that IGF-I is synthetized by cells in several tissues and organs in the adult rat. There was an apparent association between the localization of IGF-I and cell differentiation.(ABSTRACT TRUNCATED AT 250 WORDS)

Observation of inositol pentakis- and hexakis-phosphates in mammalian tissues by 31P NMR.

In analyzing the 31P NMR spectra of extracts of mammalian tissues and cells we have identified inositol pentakis- and hexakis-phosphates in essentially all of the samples examined. These compounds were present at concentrations of at least 5-15 microM. While the sources and functions of these compounds in mammalian cells are not clear, they may play an important role in phosphoinositol metabolism. For example, one obvious possibility is that these compounds may be sources of or sinks for the Ca++ mobilizing inositol tris- and tetrakis-phosphates.

Analysis of bovine udder, plate and viscera using near infrared reflectance spectroscopy.

The objective of this study was to evaluate near infrared reflectance spectroscopy (NIRS) as an accurate and inexpensive alternative to conventional chemical analyses of nonconsumer bovine tissue. Udder, plate and visceral samples were collected from mature, Charolais-Angus and Hereford-Angus crossbred beef cows at slaughter, ground and analyzed for concentrations of lipid, protein and dry matter using standard AOAC chemical procedures. Samples were analyzed using NIRS. The collection of samples was randomly split into separate calibration and validation sets. Nine calibration equations representing each constituent and tissue combination were developed, using first- or second-order derivative mathematical transformations, and those calibration equations were validated. Correlation coefficients of calibration (R) and validation (r) ranged from .95 to .98 and from .87 to .97, respectively, for all analyses except plate dry matter (r = .77). Standard errors of calibration and prediction ranged from 1.89 to 5.81%. Results from this study are interpreted to indicate that bovine udder, plate and visceral tissue composition can be accurately, quickly and efficiently predicted using NIRS technology.

Distribution of immunoreactive glia maturation factor-like molecule in organs and tissues.

Using the monoclonal antibody G2-09 raised against bovine glia maturation factor (GMF), we screened various rat organs and tissues for GMF-like immunoreactivity. In the adult animal, with the exception of the heart, GMF was found exclusively in the nervous system, with the cerebellum exhibiting higher specific activity than other brain regions. The nature of the immunoactivity in the heart is presently unclear. None of the body fluids collected from humans, including serum and cerebrospinal fluid, possessed detectable GMF immunoactivity. A phylogenetic comparison revealed the presence of GMF in the brain of al vertebrates studied, from fish to primates. GMF was absent from bacteria and yeast. An ontogenetic study on rats showed the highest GMF level in the fetal brain, with a gradual but steady decrease after birth. However, a substantial amount of GMF persisted even in older animals. GMF was localized in astrocytes and Bergmann glia in the rat brain, using immunostaining at the light microscopic level.

Iatrogenic generalized storage of dextran in patients on regular dialysis.

Intensive generalized intracellular storage of PAS positive birefringent material with an unusual ultrastructure was found in 17 patients deceased after long-term haemodialysis. Both direct and indirect evidence documented intracellular storage and "condensation" of dextran administered repeatedly for hypotension during dialysis. Intensity of storage was related to residual diuresis of the patients and not to the number of dialyses. Identification of the material was enabled by digestion with dextranase. Dextran cytoplasmatic inclusions in macrophages resisted to common solvents but dissolved in alkalies. The storage occurred not only in lymph nodes, but also in interstitial tissues of other organs, such as the heart, adrenals, bone marrow and others. The relation of massive dextran storage to frequent complications was discussed. Quantity, morphogenesis and location of storage are instructive with respect to the common knowledge of storing systems.

Cellular distribution and hormonal regulation of h-SBP in human hepatoma cells.

The cellular distribution of human Sex Steroid Binding Plasma Protein (h-SBP) was studied in human cells and tissues by indirect immunofluorescence. h-SBP was detected in the cytoplasm of hepatocytes, of prostate and epididymis epithelial cells and in endometrium. Sexual and non-sexual skin, intestine epithelium, striated muscle and some rodent organs were not labelled. The intracellular localization of h-SBP indicate that h-SBP could be taken up from the extracellular compartment or synthesized in situ in sex steroid target organs, where it may play a role in hormone uptake. The hormonal regulation of h-SBP secretion by a human hepatoma cell line, H5A, showed that tri-iodothyronine was more potent than estradiol or tamoxifen, which acted as estrogen agonist, in increasing secreted h-SBP and the combined effect of both thyroid and estrogen hormones resulted in an additive stimulation of h-SBP secretion. As shown by Northern blot analysis, oligonucleotides synthesized from the known sequence of h-SBP hybridized with a RNA of approximately 2 kb which was more represented in H5A cells than in normal human liver, and was increased 2-3 times after hormonal stimulation of the cells. The presence of a poly(A+)RNA coding for h-SBP in the human liver indicated the hepatic synthesis of this protein.

Immunohistochemical heterogeneity of alpha-tubulin in human epithelia revealed with monoclonal antibodies.

Four monoclonal antibodies raised to alpha subunit of pig brain tubulin (TU-01, TU-02, TU-03, TU-04) were used to study immunohistochemical heterogeneity of alpha tubulin in human epithelia. Selective reactivity was detected in the skin and trachea/bronchi, whereas all other epithelia investigated reacted uniformly with all four monoclonal antibodies. In the skin TU-01 reacted very strongly with all layers except the basal layer; TU-02 reacted strongly with granular layer and was unreactive or only weakly reactive with others; TU-03 reacted very strongly with basal layer and weakly to moderately with superficial layers; TU-04 reacted strongly with the granular layer of epidermis and was unreactive with other layers. In the trachea and major bronchi TU-01 reacted with the entire epithelial layer; TU-02 reacted only with superficial layer; TU-03 reacted with superficial and basal layer; TU-04 reacted only with superficial layer. Different staining patterns obtained with these four monoclonal antibodies indicate that there is immunohistochemical heterogeneity of alpha tubulin in some but not all normal human epithelia.