The effects of endogenous proteases within abdominal muscle parts on the rheological properties of thermally induced gels from white croaker (Pennahia argentata).

Three types of material meats were prepared from a so-called normal muscle part of white croaker (Pennahia argentata) containing 0, 4.2 and 8.4% of an abdominal muscle part. Thermally induced gels were then prepared from these materials by pre-heating at 65 °C for 30 or 60 min and subsequent heating at 85 °C for 20 min. The breaking strength and breaking strain rate of thermally induced gels decreased with increasing contamination levels of the abdominal muscle part, in which degradation of myosin heavy chains was observed. The proteolytic activity in the abdominal muscle part homogenate was highest at 62.5 °C. These results suggest that the abdominal muscle part contains proteases that induce the modori phenomenon. Technical experts assume that a contaminated abdominal muscle part leads to quality deterioration in surimi production industries. Our findings will aid the production of high-quality surimi-based products.

Altered Integrins Expression of Patients Affected by Cryptorchidism.

OBJECTIVES: The study aimed to investigate the expression of the integrin isoforms α7A and ß1A, expressed by myogenic precursor cells, and α7B and ß1D, expressed by mature muscle cells in the cremaster of patients affected by an undescended testis. METHODS: Fifteen samples of cremaster were obtained from patients undergoing surgery for an undescended testis. Thirty control specimens of cremaster were harvested from patients with congenital hydrocele or inguinal hernia. Immunofluorescent analysis was carried out using anti-α7A, ß1A, α7B, and ß1D integrin antibodies. Sections were observed using confocal laser scanning microscopy. RESULTS: As compared with controls, a significant loss of a α7B (p = 0.0355) and ß1D (p = 0.0069) integrins and a higher expression of α7A (p = 0.0003) and ß1A (p = 0.0150) was detected in the cremaster of patients affected by an undescended testis. CONCLUSIONS: Our data document a critical alteration of the cytoskeleton of cremasteric smooth muscle cells in patients with an undescended testis. This might explain the altered function in smooth muscle cells in cremaster implied during testicular descent. We therefore speculate that the postnatal splicing of α7A to α7B and of ß1A to ß1D integrins is delayed. This could account for the common clinical scenario of spontaneous descent of the testes in the first months of life.

Sodium storage in human tissues is mediated by glycosaminoglycan expression.

The current paradigm regarding sodium handling in animals and humans postulates that total body sodium is regulated predominately via regulation of extracellular volume. Active sodium storage independent of volume retention is thought to be negligible. However, studies in animals, hypertensive patients, and healthy humans suggest water-free storage of sodium in skin. We hypothesized that tissue sodium concentrations ([Na]T) found in humans vary and reflect regulation due to variable glycosaminoglycan content due to variable expression of XYLT-1. Twenty seven patients on dialysis and 21 living kidney transplant donors free of clinically detectable edema were studied. During surgery, abdominal skin, muscle, and arteries were biopsied. [Na]T was determined by inductively coupled plasma-optical emission spectrometry, semiquantitative glycosaminoglycan content with Alcian stain, and XYLT-1 expression by real-time PCR. [Na]T of arteries were ranging between 0.86 and 9.83 g/kg wet wt and were significantly higher in arteries (4.52 ± 1.82 g/kg) than in muscle (2.03 ± 1.41 g/kg; P < 0.001) or skin (3.24 ± 2.26 g/kg wet wt; P = 0.038). For individual patients [Na]T correlated for skin and arterial tissue (r = 0.440, P = 0.012). [Na]T also correlated significantly with blinded semiquantitative analysis of glycosaminoglycans staining (r = 0.588, P = 0.004). In arteries XYLT-1 expression was also correlated with [Na]T (r = 0.392, P = 0.003). Our data confirm highly variable [Na]T in human skin and muscle and extend this observation to [Na]T in human arteries. These data support the hypothesis of water-independent sodium storage via regulated glycosaminoglycan synthesis in human tissues, including arteries.

Influence of tobacco, alcohol and diabetes on the collagen of cremaster muscle in patients with inguinal hernias / Influência do tabaco, álcool e diabete sobre a interação célula-colágeno em músculo cremaster de humanos com hérnias inguinais

ABSTRACT Background: New findings point out that the mechanism of formation of the hernias can be related to the collagenous tissues, under activity of aggressive agents such as the tobacco, alcohol and diabetes. Aim: To analyze the collagen present in the cremaster muscle in patients with inguinal hernias, focusing the effect of tobacco, alcohol, and diabetes. Methods: Fifteen patients with inguinal hernia divided in three groups were studied: group I (n=5) was control; group II (n=5) were smokers and/or drinkers; and group III (n=5) had diabetes mellitus. All subjects were underwent to surgical repair of the inguinal hernias obeying the same pre, intra and postoperative conditions. During surgery, samples of the cremaster muscle were collected for analysis in polarized light microscopy, collagen morphometry and protein. Results: The area occupied by the connective tissue was higher in groups II and III (p<0.05). The collagen tissue occupied the majority of the samples analyzed in comparison to the area occupied by muscle cells. The content of total protein was higher in groups II and III compared to the control group (p<0.05). Conclusion: The tobacco, alcohol and diabetes cause a remodel the cremaster muscle, leading to a loss of support or structural change in this region, which may enhance the occurrences and damage related to inguinal hernias.

RESUMO Racional: Estudos recentes sinalizam que o mecanismo de formação das hérnias pode estar relacionado aos tecidos colagenosos, sob a ação de agentes agressores como o tabaco, o álcool e o diabete. Objetivo: Avaliar o colágeno presente no músculo cremaster em pacientes com hérnias inguinais enfocando o efeito do tabaco, álcool e diabete. Métodos: Foram estudados 15 pacientes com hérnias inguinais divididos em: grupo I (n=5) controles; grupo II (n=5) indivíduos fumantes e/ou etilistas; e grupo III (n=5) indivíduos que apresentavam diabete melito. Todos foram submetidos à correção cirúrgica das hérnias inguinais obedecendo às mesmas condições pré, intra e pós-operatórias. Durante o procedimento cirúrgico, amostras do músculo cremaster foram coletadas para análises em microscopia de luz polarizada, morfometria do colágeno e de proteínas. Resultados: A área ocupada por tecido conjuntivo foi maior nos grupos II e III (p<0,05). O tecido colágeno ocupou a maior parte das amostras analisadas, em comparação à área ocupada pelas células musculares. O conteúdo de proteínas totais foi maior nos grupos II e III, quando comparado com o grupo controle (p<0,05). Conclusão: O tabaco, o álcool e o diabete ocasionam remodelação no músculo cremaster, levando à perda de suporte ou alteração estrutural nesta região, podendo intensificar as ocorrências e os danos relacionados às hérnias inguinais.

INFLUENCE OF TOBACCO, ALCOHOL AND DIABETES ON THE COLLAGEN OF CREMASTER MUSCLE IN PATIENTS WITH INGUINAL HERNIAS.

Background: New findings point out that the mechanism of formation of the hernias can be related to the collagenous tissues, under activity of aggressive agents such as the tobacco, alcohol and diabetes. Aim: To analyze the collagen present in the cremaster muscle in patients with inguinal hernias, focusing the effect of tobacco, alcohol, and diabetes. Methods: Fifteen patients with inguinal hernia divided in three groups were studied: group I (n=5) was control; group II (n=5) were smokers and/or drinkers; and group III (n=5) had diabetes mellitus. All subjects were underwent to surgical repair of the inguinal hernias obeying the same pre, intra and postoperative conditions. During surgery, samples of the cremaster muscle were collected for analysis in polarized light microscopy, collagen morphometry and protein. Results: The area occupied by the connective tissue was higher in groups II and III (p<0.05). The collagen tissue occupied the majority of the samples analyzed in comparison to the area occupied by muscle cells. The content of total protein was higher in groups II and III compared to the control group (p<0.05). Conclusion: The tobacco, alcohol and diabetes cause a remodel the cremaster muscle, leading to a loss of support or structural change in this region, which may enhance the occurrences and damage related to inguinal hernias.

Racional: Estudos recentes sinalizam que o mecanismo de formação das hérnias pode estar relacionado aos tecidos colagenosos, sob a ação de agentes agressores como o tabaco, o álcool e o diabete. Objetivo: Avaliar o colágeno presente no músculo cremaster em pacientes com hérnias inguinais enfocando o efeito do tabaco, álcool e diabete. Métodos: Foram estudados 15 pacientes com hérnias inguinais divididos em: grupo I (n=5) controles; grupo II (n=5) indivíduos fumantes e/ou etilistas; e grupo III (n=5) indivíduos que apresentavam diabete melito. Todos foram submetidos à correção cirúrgica das hérnias inguinais obedecendo às mesmas condições pré, intra e pós-operatórias. Durante o procedimento cirúrgico, amostras do músculo cremaster foram coletadas para análises em microscopia de luz polarizada, morfometria do colágeno e de proteínas. Resultados: A área ocupada por tecido conjuntivo foi maior nos grupos II e III (p<0,05). O tecido colágeno ocupou a maior parte das amostras analisadas, em comparação à área ocupada pelas células musculares. O conteúdo de proteínas totais foi maior nos grupos II e III, quando comparado com o grupo controle (p<0,05). Conclusão: O tabaco, o álcool e o diabete ocasionam remodelação no músculo cremaster, levando à perda de suporte ou alteração estrutural nesta região, podendo intensificar as ocorrências e os danos relacionados às hérnias inguinais.

Factors affecting the tissues composition of pork belly.

Bellies derived from the commercial population of pig carcasses are diverse in terms of tissue composition. Knowledge of the factors influencing it and the expected results, permits quick and easy evaluation of raw material. The study was designed to determine the factors affecting the tissues composition of pork bellies and to estimate their lean meat content. The research population (n=140 pig carcasses) was divided into groups according to sex (gilts, barrows), half-carcass mass (<40, 40 to 43.9, 44 to 46.9, ⩾47 kg) and lean meat content class: S (⩾60%), E (55% to 60%), U (50% to 55%), R (<50%). Bellies were subjected to a detailed dissection. Half-carcass mass affected the levels of all the analysed parameters. The only exception was the mass of the fat with the skin in the 40 to 43.9 kg group, for which the value did not differ statistically between the two groups <40 and 44 to 46.9 kg. Decrease in lean meat content affected the growth of the fat and skin mass in a linear way. No differences were observed between class S and E in terms of belly muscle mass. A 0.37% higher share of belly in the half-carcass was found for barrows (P<0.001), although bellies issued from barrows were characterized by a higher proportion of fat with skin compared with gilts (P=0.02). Interactions were observed between sex and half-carcass mass, so the sex of heavy half-carcasses becomes an important determinant for conditioning the muscle content. Equations were calculated and allow a fast and highly accurate determination of the lean meat content in bellies, suggesting they may be used directly in the production line.

The occurrence of two types of fast skeletal myosin heavy chains from abdominal muscle of kuruma shrimp Marsupenaeus japonicus and their different tissue distribution.

Shrimps belong to the class Crustacea, which forms a large, diverse group in the invertebrates. However, the physiology and biochemistry of their skeletal muscles have been poorly understood compared with those from vertebrates including mammals and fish. The present study focused on myosin, the major protein in skeletal muscle, from adult specimens of kuruma shrimp Marsupenaeus japonicus. Two types of the gene encoding myosin heavy chain (MHC), a large subunit of the myosin molecule, were cloned from abdominal fast skeletal muscle and defined as MHCa and MHCb. Protein analysis revealed that the MHCa isoform was expressed at a higher level than the MHCb isoform. The full-length cDNA clones of MHCa and MHCb consisted of 5929 bp and 5955 bp, respectively, which encoded 1912 and 1910 amino acids, respectively. Both were classified into fast muscle type by comparison with the partially deduced amino acid sequences of fast-type and slow-type (S(1), slow twitch) MHCs reported previously for the American lobster Homarus americanus. The amino acid identities between MHCa and MHCb of kuruma shrimp were 78%, 60% and 72% in the regions of subfragment-1, subfragment-2 and light meromyosin, respectively, and 71% in total. In situ hybridisation using anti-sense RNA-specific probes, along with northern blot analysis using different tissues from abdominal muscle, revealed the different localisation of MHCa and MHCb transcripts in abdominal fast skeletal muscle, suggesting their distinct physiological functions.

Abdominal vagal afferent pathways and their distributions of intraganglionic laminar endings in the rat duodenum.

We examined abdominal vagal afferents (n = 33) and the distributions of their intraganglionic laminar endings (IGLEs) in the duodenum. Rats (male, Sprague-Dawley) received a partial subdiaphragmatic vagotomy that spared a single branch. Wheat germ agglutinin-horseradish peroxidase (0.5-1.0 µl) was injected into the nodose ganglion ipsilateral to the vagotomized side. We observed that the hepatic branch does not project to the stomach, that the accessory celiac and celiac branches course along the celiac artery and innervate the intestines, and that the left nodose afferents innervate predominantly the duodenum. The hepatic branch innervates the duodenum via the "hepatoduodenal" subbranch and has the densest IGLE distribution in both the dorsoventral and the rostrocaudal extensions of the first 4-cm segment. Both gastric branches have two subbranches that innervate the duodenum; the "lesser curvature" subbranches follow the lesser curvature artery and may join the "hepatoduodenal" subbranch, whereas the "pyloric" subbranches run through the antrum and pylorus to reach the proximal duodenum. Moreover, the subbranches of ventral and dorsal gastric branches innervate more in the ventral and dorsal parts of the duodenum, respectively, and have more IGLEs in the rostral region than in the caudal. A posteriori comparisons indicate that, in the first-centimeter segment, the ventral gastric branch has significantly more IGLEs, whereas, in the third- and fourth-centimeter segments, the hepatic branch has more IGLEs. The finding that three different vagal branches innervate the duodenum with different densities of afferent endings might indicate a viscerotopic receptive field that coordinates digestive functions in feeding.

Effects of L-carnitine supplementation on quality characteristics of fresh pork bellies from pigs fed 3 levels of corn oil.

Crossbred pigs (n = 216) were used to test the effect of supplemental L-carnitine (CARN) on the fatty acid composition and quality characteristics of fresh pork bellies from pigs fed diets formulated with different inclusion levels of corn oil. Pigs were blocked by BW (43.6 ± 1.0 kg) and allotted randomly to pens of 6 pigs within blocks. Then, within blocks, pens were assigned randomly to 1 of 6 dietary treatments in a 2 × 3 factorial arrangement, with either 0 or 100 mg/kg of supplemental CARN and 3 dietary inclusion levels (0, 2, or 4%) of corn oil (CO). When the lightest block weighed 125.0 kg, all pigs were slaughtered, and left-side bellies were captured during carcass fabrication for quality data collection. Fresh pork bellies were evaluated for length, width, thickness, and firmness (bar-suspension and Instron-compression methods) before a 2.5-cm-wide strip of belly was removed and subsequently dissected into subcutaneous fat, primary lean (latissimus dorsi), secondary lean (cutaneous trunci), and intermuscular fat for fatty acid composition determination. Although belly length, width, and thickness of fresh pork bellies were not affected by CARN (P ≥ 0.128) or CO (P ≥ 0.073), belly firmness decreased linearly (P < 0.001) with increasing dietary CO, but there was no (P ≥ 0.137) effect of CARN on any belly firmness measure. Dietary CARN increased (P < 0.05) the proportion of total SFA in the intermuscular fat layer, increased (P < 0.05) the proportion of total MUFA in the primary and secondary lean layers, and decreased (P < 0.05) the proportion of total PUFA in the intermuscular fat and secondary lean layers of pork bellies. Moreover, the SFA and MUFA compositions decreased linearly (P < 0.001) with increasing dietary CO, and the rate of the decrease in SFA composition was greater (P < 0.001) in the fat layers than the lean layers. Conversely, the PUFA content increased linearly (P < 0.001) with increasing dietary CO, and the rate of the increase in PUFA was greater (P < 0.001) in the fat than the lean layers, and greater (P = 0.022) in the primary than secondary lean layer. Results from this study would indicate that differences in the amount and rate of fatty acid deposition associated with feeding increased amounts of CO, along with moisture differences among the belly layers, combine to negatively affect fresh pork belly firmness.

Compositional and instrumental firmness variations within fresh pork bellies.

Fresh pork bellies (n=24) were cut into 15 sections to measure the intra-belly variation in compositional and mechanical firmness characteristics. Length and width of each belly was measured before the belly was divided into 3 rows (D = dorsal; C = central; and V = ventral) and 5 columns (labeled 1, 2, 3, 4, and 5 from cranial to caudal), resulting in 15 belly sections of equal dimensions. The belly section with the greatest compression value was D-1, whereas the lowest compression value was found in the V-4 section (column×row, P<0.001). Conversely, the greatest and least puncture values were observed in the C-2 and V-5 locations, respectively (column×row, P=0.016). The D-3 section had the lowest proportion of lean and the greatest proportion of fat, but the greatest lean and lowest fat percentages were found in the V-1 and C-4 sections, respectively (column×row, P<0.001). The greatest proportions of saturated fatty acids (SFA) were found in the V-4 and V-5, and the lowest proportions of SFA were in D-1 (column×row, P<0.001). Moreover, C-4 and V-1 had the greatest percentages of monounsaturated fatty acids (MUFA), whereas the lowest MUFA content was observed in D-1, D-2, and D-3 (column×row, P<0.001). The D row (columns 1, 2, 3, and 5) also had the greatest proportion of polyunsaturated fatty acids (PUFA), but the lowest proportions of PUFA were located in C-4, V-4, and V-5 (column×row, P<0.001). Consequently, the iodine value was greatest in D-1 and lowest in V-4, V-5, and C-5 (column×row, P<0.001). It is apparent from these results that there is an obvious fatty acid composition gradient within bellies, which results in considerable intra-belly variation in composition and firmness.

The specific binding sites of eyestalk- and pericardial organ-crustacean hyperglycaemic hormones (CHHs) in multiple tissues of the blue crab, Callinectes sapidus.

Crustacean hyperglycaemic hormone from the pericardial organ (PO-CHH) is a CHH-related neuropeptide but its function and target tissues are not known in crustaceans. To investigate this issue, we employed radiolabelled ligand binding and cGMP assays, using eyestalk-CHH (ES-CHH) as a reference neuropeptide. The membranes were prepared from various tissues of Callinectes sapidus: hepatopancreas, hindgut, midgut, gills, heart, abdominal muscles and scaphognathites. Like ES-CHH, recombinant PO-CHH (rPO-CHH) specifically bound to the membranes of scaphognathites=abdominal muscles>midgut>gills> heart>hindgut and hepatopancreas (list order corresponds to the number of binding sites). The specific binding sites of (125)I-ES-CHH in hepatopancreas and gills were saturable and displaceable. The abdominal muscle membrane binding sites were specific and saturable to both CHHs. These binding sites were displaced by homologous neuropeptides, but poorly displaced by the heterologous counterpart. As for the second messenger, the expected increment (3- to >20-fold) in the amount of cGMP produced by ES-CHH was noted in most tissues tested except midgut. Recombinant PO-CHH increased cGMP production 1.5- to 4-fold in scaphognathites, heart, midgut, hindgut and abdominal muscles. The results obtained from the binding study suggest that PO-CHH also has multiple target tissues of which abdominal muscles and scaphognathites are the primary ones. The differences in the primary amino acid sequences of PO-CHH and ES-CHH, particularly in the C-terminal region and in the amidation at C-terminus, may contribute to the truncated responses of hyperglycaemia, cGMP stimulation and binding affinity.

Contents of selected heavy metals in the liver, kidneys and abdominal muscle of the brown hare (Lepus europaeus Pallas, 1778) in Central Pomerania, Poland.

The content of iron, zinc, copper, manganese, lead, and cadmium was determined in the kidney, liver, and abdominal muscle of the brown hare (Lepus europaeus), a species known for its bioindicative potential in environmental quality assessment. The animals assayed were divided into immature (in their first year of life; n=25) and adult (n=39). The hares were acquired in an area situated far away from major cities and direct impacts of industrial and transport pollution. The zinc content found in the assayed hare's muscles (25.9 microg/g w.w.) and that of manganese in the kidneys, liver, and muscles (2, 2.51 i 0.85 microg/g w.w., respectively) tended to be higher than those reported from the brown hare elsewhere in Europe. The content of cadmium (particularly in the kidneys) and lead proved substantial and close to those typical of the hare inhabiting industry-affected areas. Analysis of the data pooled for all the individuals showed Zn and Cd contents to increase with age in the kidneys, the liver Cd content increasing with age as well; on the other hand, the age-Cu content was negative. In addition, a number of significant correlations between the metals themselves were revealed, particularly with respect to Zn-Cd correlation in kidneys (r(s)=0.47), Fe-Cd, Zn-Cu, Mn-Zn, Mn-Cu, and Mn-Pb in the liver (r(s) of 0.42, 0.86, 0.72, 0.79, and 0.42, respectively), and Zn-Cu in the muscle (r(s)=0.56). The kidney cadmium content was higher by 81% in the adult than in the immature hare, the adult hare muscle copper content being lower by 15.5%.

Effects of pH and molecular charge on dipolar coupling interactions of solutes in skeletal muscle observed by DQF, 1H NMR spectroscopy.

In this study we tested the effect of molecular charge and chirality as well as tissue pH on dipolar coupling interaction in skeletal muscle. These results were demonstrated by double quantum filtered, DQF, 1H NMR spectra acquired on permeable skeletal muscle samples dialyzed against buffered solutions containing three classes of solutes-electrolytes (lactate and Tris), zwitterions (alanine and glycine), and non-electrolytes (dioxane and ethanol)-as a function of pH ranging from 5.0 to 8.5. The results show that charge density on the protein filaments strongly influences dipolar coupling of solutes in muscle whereas charge on the solutes themselves has only a small effect. The frequency splitting of the dipolar coupled peaks for all the molecules tested was strongly affected by muscle pH. Higher pH increased negative charge density on the filaments and resulted in weaker dipolar coupling for anions and zwitterions but stronger coupling for the cation TRIS. Molecular charge per se or chirality did not affect the frequency splitting of the dipolar coupled peaks. The molecules, lactate, ethanol, and alanine, have scalar coupled spins and consequently a double quantum signal in solution. However, spectra acquired from these molecules in muscle showed an additional frequency splitting due to additional dipolar coupling interactions. Due to lack of scalar coupling, spectra from Tris, glycine, and dioxane showed no double quantum signal in solution but did when in muscle. All these observations can be explained by the fact that the net charge on protein filaments dominates the mechanism of dipolar coupling interactions in the highly anisotropic structures in muscle.

Actinomicosis primaria de pared abdominal con peritonitis secundaria / Primary abdominal wall actinomycosis with secondary peritonitis

La actinomicosis es una entidad causada por el Actinomyces israelii que se presenta sobre todo en la zona cervicofacial, abdominal y torácica. En la región abdominal se transmite a partir de zonas de lesión tisular, siendo lo más frecuente la aparición de clínica en el hemiabdomen derecho, a pesar de existir una relativa tendencia en la actualidad hacia la diversificación de localizaciones. Sin embargo, la aparición de un cuadro de actinomicosis primaria de pared abdominal es una entidad poco habitual, motivo por el cual presentamos nuestro caso. El tratamiento de la actinomicosis abdominal consiste en la combinación de una adecuada antibioterapia con el correspondiente desbridamiento quirúrgico (AU)

Collagen fibers in linea alba and rectus sheaths. I. General scheme and morphological aspects.

BACKGROUND: The anatomy of the anterior abdominal wall plays a most significant role in surgery. Thus the three-dimensional architecture of the collagen fibers in linea alba and rectus sheaths was investigated in 12 human cadavers. MATERIAL AND METHODS: The linea alba was divided into 14 different anatomical segments in the craniocaudal direction. Two-hundred-micrometer-thick, eosin-stained sections from these segments were analyzed by confocal laser scanning microscopy. In this way the direction of the collagen fibers was estimated in the midline of the linea alba and in the medial parts of the rectus sheaths. Width and thickness of the linea alba and thickness of the rectus sheaths were measured. RESULTS: In the ventral rectus sheath essentially oblique fibril bundles intermingle with each other, while the dorsal rectus sheath consists chiefly of transverse fibril bundles. In the linea alba three different zones of fiber orientation follow each other from ventral to dorsal: The lamina fibrae obliquae consists of intermingling oblique fibers. The lamina fibrae transversae contains mainly transverse fibril bundles, while an inconstant, small lamina fibrae irregularium is composed of oblique fibers. Different regions can be distinguished in the craniocaudal course of the linea alba: supraumbilical part, umbilical part, transition zone, and infraarcuate part. CONCLUSIONS: A new model of fiber architecture of the linea alba was developed that describes the fiber architecture as a three-dimensional, highly structured meshwork of collagen fibers. In contrast to former models, no separate lines of decussation of the fibers could be found.

Molecular species of collagen in the intramuscular connective tissues of the marine crab, Scylla serrata.

Type V like collagens are widely distributed in marine invertebrates, particularly crustaceans and molluscs. We have been investigating the nature of collagens in the muscular tissues of crustaceans. The presence of type V like homotrimeric collagen in prawn muscle was noted before. We report here a comparative analysis of collagens purified from the pepsin digest of abdominal and pereiopod muscle tissues of the crab, Scylla serrata. The major collagen in either muscle precipitated at 1.2 M NaCl at acid pH, suggestive of a type V like property. The homotrimeric collagen was then purified to near homogeneity by precipitation with 20% ammonium sulphate. Solubility characteristics and biochemical studies indicated the leg muscle collagens to be highly crosslinked and stabilised by more bound carbohydrates, as compared to the abdominal muscle collagen. Analysis of amino acid composition revealed a close similarity to known type V collagens and the leg muscle collagen was characterised by more lysine hydroxylation and slightly reduced glycine content. The leg muscle collagen had a higher denaturation temperature and intrinsic viscosity than the abdominal muscle collagen. Our results confirm the similarity of major crustacean muscle collagens to vertebrate type V collagen. Further, the relative complexity of leg muscle collagen, unlike the abdominal muscle collagen, correlates to the specific functional requirements, where the former is involved in locomotion and preying and the latter in normal growth and development.

[Tissue amino acids in patients with colorectal carcinoma]. / Tkánový aminoacidogram u nemocných s kolorektálním karcinomem.

Different catabolic conditions have their special characteristics of intracellular biochemical changes. The objective of the presented work is to assess the concentration of free amino acids in muscles of patients with colorectal carcinoma. In a group of 17 patients the free amino acid level was assessed in tissues and plasma. Material was collected on operation by biopsy of the rectus abdominis muscle, the concentration of different amino acids was assessed by the HPLCV method with fluorescent detection. For statistical evaluation the T-test was used. From the results ensues that in patients with colorectal cancer in plasma statistically significantly lower taurine, glutamine, valine, tyrosine levels were found, intracellularly significantly reduced levels of taurine, glutamic acid, methionine and ornithine were recorded. Significantly elevated intracellular levels of valine, isoleucine, leucine, tyrosine and phenylalanine were found. Assessment of the tissue aminoacidogram is the first step when attempting to influence the intracellular amino acid concentration by defined dietetic preparations.

Cloning of tropomyosins from lobster (Homarus americanus) striated muscles: fast and slow isoforms may be generated from the same transcript.

Complementary DNAs encoding fibre-type-specific isoforms of tropomyosin (Tm) have been isolated from lobster (Homarus americanus) striated muscle expression libraries made from poly(A)+ RNA purified from deep abdominal (fast-type) and crusher-claw closer (slow-type) muscles. A cDNA of slow-muscle Tm (sTm1), containing a complete open reading frame (ORF) and portions of the 5' and 3' untranslated regions (UTRs), encodes a protein of 284 amino acid residues with a predicted mass of 32,950, assuming acetylation of the amino terminus. The nucleotide sequence of a fast-muscle tropomyosin (fTm cDNA), which includes the entire ORF and part of the 3' UTR, is identical to that of sTm1 cDNA, except in the region encoding amino acid residues 39-80 (equivalent to exon 2 of mammalian and Drosophila muscle tropomyosin genes). The deduced amino acid sequences, which display the heptameric repeats of nonpolar and charged amino acids characteristic of alpha-helical coiled-coils, are highly homologous to tropomyosins from rabbit, Drosophila, and shrimp (57% to 99% identities, depending on species). Northern blot analysis showed that two transcripts (1.1 and 2.1 kb) are present in both fibre types. Mass spectrometry indicated that fast muscle contains one major isoform (fTm: 32,903), while slow muscle contains two major isoforms (sTm1 and sTm2: 32,950 and 32,884 respectively). Both Tm preparations contained minor species with a mass of about 32,830. Sequences of peptides derived from purified slow and fast Tms were identical to the deduced amino acid sequences of the sTm1 and fTm cDNAs, respectively, except in the C-terminal region of fTm. The difference in mass between that predicted by the deduced sequence (32,880) and that measured by mass spectrometry (32,903) suggests that fTm is posttranslationally modified, in addition to acetylation of the N-terminal methionine. These data are consistent with the hypothesis that the fTm and sTm1 are generated by alternative splicing of two mutually-exclusive exons near the 5' end of the same gene.

Concentrations of prophylactic ceftriaxone in abdominal tissues during pancreatic surgery.

Ceftriaxone concentrations in abdominal tissues were evaluated after administration as antibiotic prophylaxis for pancreatic surgery. Ten patients were given ceftriaxone (1 g i.v.) 30 min before surgery. Ceftriaxone concentrations in fatty tissues ranged from 2.5 to 6.2 microg/g. Ceftriaxone concentrations were 6.0 +/- 8.6 microg/g in pancreatic tissues, 2.1 +/- 2.5 mg/L in pancreatic fluid, 1179 +/- 1271 mg/L in pancreatic bile, and 18 +/- 16 microg/g in the liver. In fatty tissues, 8-10 patients had tissue levels greater than the MIC90 for Staphylococcus aureus and Escherichia coli and the 10 patients had tissue levels greater than the MIC90 for Klebsiella pneumoniae and Proteus mirabilis. In other tissues, penetration was greater than the MIC90 for potential pathogens in 50-100% of the patients.

Submucosal collagen in experimental gastroschisis.

Gastroschisis is a congenital anomaly in which the intestines are exposed to amniotic fluid throughout fetal life. Previous studies in animal models have demonstrated smooth muscle thickening and decreased contractility, epithelial dysfunction, and submucosal thickening. The present studies were done to further define the mechanism of submucosal changes by investigating collagen deposition and gene expression in a rabbit model. Gastroschisis was surgically created in fetal rabbits at 24 days gestation (term is 31 days). Sham-operated and unoperated fetuses served as controls. Fetuses were sacrificed and bowels were harvested at 26, 28, and 31 days gestation. Animal weight and gross and histologic appearance were assessed. Submucosal collagen content was measured using the van Geison stain. In situ hybridization of the expression of alpha (1) procollagen RNA was done to determine the distribution and source of submucosal collagen. At term, submucosal thickening was present in animals with gastroschisis, associated with a significantly increased collagen content. Collagen distribution was also more diffuse in the gastroschisis animals than in controls. In situ hybridization revealed procollagen expression in round cells located in the submucosa and not in smooth muscle. These cells did not resemble fibroblasts, and their identity is uncertain. Experimental gastroschisis is characterized by submucosal thickening which is associated with changes in collagen, including increased deposition and more diffuse distribution in the submucosa. The cells responsible for production of procollagen are round, nonfibroblast cells which are located in the submucosa and not in the smooth muscle layer. These findings may have some importance in understanding the mechanisms responsible for intestinal malfunction in infants with gastroschisis.