Human Psoas Muscle Research Articles

Comparative histochemical and morphometric analysis of muscle fibers of the psoas muscle in individuals of both genders with ageing

Background/Aim. There is a reduction of the psoas muscle with aging, and histopathological analysis (HPA) on postmortem material also shows its significant changes with advancing age. The aim of this study was to determine the presence and distribution of types I and II muscle fibers in the human psoas muscle in individuals of different ages and genders. Methods. The material consisted of tissue samples of the right iliopsoas muscle taken from 30 adult cadavers (18 males and 12 females), aged from 30 to 90 years, divided into three age groups. The material was obtained from the Institute of Forensic Medicine, Faculty of Medicine, University of Nis, Serbia. Hematoxylin and eosin (H&E) staining was used in the H PA of muscle cells. Results. The values of astereological parameters (area, perimeter, and Feret's diameter) of type I and type II muscle fibers were greater in male cases in comparison with female ones, although with no statistically significant difference. Based on the histochemical and morphometric analysis, it was concluded that, after 70 years of life, a loss of type II muscle fibers occurred, which was more conspicuous in female cases. Conclusion. During aging, the loss of type II muscle fibers, as well as the atrophy of type I and type II muscle fibers, demonstrate similar dynamics in both genders.

Long-term room temperature preservation of corpse soft tissue: an approach for tissue sample storage

BackgroundDisaster victim identification (DVI) represents one of the most difficult challenges in forensic sciences, and subsequent DNA typing is essential. Collected samples for DNA-based human identification are usually stored at low temperature to halt the degradation processes of human remains. We have developed a simple and reliable procedure for soft tissue storage and preservation for DNA extraction. It ensures high quality DNA suitable for PCR-based DNA typing after at least 1 year of room temperature storage.MethodsFragments of human psoas muscle were exposed to three different environmental conditions for diverse time periods at room temperature. Storage conditions included: (a) a preserving medium consisting of solid sodium chloride (salt), (b) no additional substances and (c) garden soil. DNA was extracted with proteinase K/SDS followed by organic solvent treatment and concentration by centrifugal filter devices. Quantification was carried out by real-time PCR using commercial kits. Short tandem repeat (STR) typing profiles were analysed with 'expert software'.ResultsDNA quantities recovered from samples stored in salt were similar up to the complete storage time and underscored the effectiveness of the preservation method. It was possible to reliably and accurately type different genetic systems including autosomal STRs and mitochondrial and Y-chromosome haplogroups. Autosomal STR typing quality was evaluated by expert software, denoting high quality profiles from DNA samples obtained from corpse tissue stored in salt for up to 365 days.ConclusionsThe procedure proposed herein is a cost efficient alternative for storage of human remains in challenging environmental areas, such as mass disaster locations, mass graves and exhumations. This technique should be considered as an additional method for sample storage when preservation of DNA integrity is required for PCR-based DNA typing.

Localization of motor nerve branches of the human psoas muscle

Endplate-targeted botulinum toxin injections can achieve optimal neuromuscular blockade. The goal of this study was to identify the motor endplate (MEP) zone in the human psoas muscle through dissection. In 24 human cadaver psoas muscles the nerve branches and their intramuscular course were followed by stereoscopic microscopic dissection as far as their terminal ramifications. From the lumbar plexus, an average of 3.7 (range 2-7) nerve branches enter the psoas muscle. The proximal and distal limit of the MEP-zone are situated at about 30% and 70%, respectively, of the distance between the twelfth thoracic vertebra (Th12) and the passing of the psoas under the inguinal ligament. In reference to the sacral promontory (P), these limits are respectively from 50% of the Th12-P distance to 20% of the P-pubis distance. This study of the MEP zone of the human psoas muscle can allow the clinician to inject BTX-A close to its site of action.

Regression of polyneural innervation in the human psoas muscle.

During the early stages of mammalian ontogeny muscle fibres are innervated by more than one axon. This polyneural innervation is replaced by mononeural innervation in the course of development. The regression of polyneural innervation in the psoas muscle in the human is the topic of the present study. Innervation patterns were studied in fetuses from 15 1/2 weeks of post menstrual age (PMA) and in babies until 80 weeks PMA (40 weeks after term age) and compared to data from two adults. Motor endplates were stained by a combined acetylcholinesterase stain. Innervation patterns and motor endplate morphology were studied and the sizes of endplates were measured. As a main result of our study polyneural innervation of the psoas muscle remains at a level of about 2 endings per endplate (range 1-5 terminals) until 18-25 weeks PMA and decreases thereafter. From 52 weeks PMA (12 weeks post term) onwards, muscle fibres are predominantly mononeurally innervated. During development the morphology of the terminal patterns of the nerve endings becomes more complex and the size of endplates increases, implying that the adult pattern of muscle innervation is reached at the age at which a major functional transformation in the neurobehavioural repertoire occurs (i.e. the end of the second and the beginning of the third month.

Tissue dosimetry of 632 nm light delivered through brachytherapy needles

It would be useful to be able to deliver both interstitial photodynamic therapy (PDT) and brachytherapy with the same applicator system. At present, data on the attenuation of PDT-range light in human tissues are scarce and there are no data using brachytherapy needles. The purpose of this project is to evaluate the effective attenuation coefficient (μeff) of 632 nm light in human ex vivo muscle, liver, and prostate specimens with the light delivered through the type of needles that are commonly used to deliver interstitial brachytherapy. Specimens of human psoas muscle, liver, and prostate were obtained at autopsy. Light (632 nm) was delivered to the tissues and collected (measured) with standard optical fibers placed inside the type of plastic needles that are commonly used for interstitial brachytherapy. Light intensity, as a function of the distance between source and collection fiber, was measured at 0.1 mm increments along the catheter tract that was perpendicular to the light source. The μeffs for human muscle, liver, and prostate were 0.27, 1.52, and 0.17 mm−1. Radiat. Oncol. Invest. 4:281–286, 1996. © 1997 Wiley-Liss, Inc.

Quantification of carbonic anhydrase III and myoglobin in different fiber types of human psoas muscle.

Carbonic anhydrase (CA III) and myoglobin contents from isolated human muscle fibers were quantified using a sensitive time-resolved fluoroimmunoassay. Human psoas muscle specimens were freeze-dried, and single fibers were dissected out and classified into type I, IIA and IIB by myosin ATPase staining. Fiber typing was further confirmed by SDS-PAGE. CA III and myoglobin were found in all fiber types. Type I fibers contained higher concentrations of CA III and myoglobin than type IIA and IIB fibers. The relative concentrations of CA III in type IIA and IIB fibers were respectively 24% and 10% of that in type I fibers. The relative concentrations of myoglobin in type IIA and IIB fibers were 60% and 28% of that in type I fibers. Anti-CA III immunoblotting results from fiber-specific pooled samples agreed well with quantitative measurements. The results indicate that CA III is a more specific marker than myoglobin for type I fibers.

Postmortem interval estimation by creatinine levels in human psoas muscle

A new method was tested for estimating time after death in the middle range of postmortem interval (weeks). Creatinine muscular concentration in human cadaver is positively correlated to postmortem estimation. Temperature should be mainly related to the creatinine transformation rate. A stronger correlation was found at 11°C up to 30 days and at 20°Celsius up to 15 days.

Cloning and expression of human aldose reductase.

The complete amino acid sequence of human retina and muscle aldose reductase was determined by nucleotide analysis of cDNA clones isolated using synthetic oligonucleotide probes based on partial amino acid sequences of purified human psoas muscle aldose reductase. The cDNA sequence differs substantially in the noncoding and coding regions of recently published sequences of this enzyme. The mRNA for aldose reductase was abundantly expressed in HeLa cells, but only scarcely in a neuroblastoma cell line. Recombinant baculovirus containing one of the muscle cDNA clones was constructed and used to infect Spodoptera frugiperda (SF9) cells. A prominent protein with an apparent molecular size of 36 kDa was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the culture medium as well as in the homogenate of SF9 cells after 2 days of infection. Culture medium or the supernatant fraction of cell homogenates containing this protein had high aldose reductase activity which showed characteristics of the reported human enzyme. These findings indicate that the amino acid sequence reported in this paper represents human retina and muscle aldose reductase and that functional human aldose reductase can be expressed in large amounts in a baculovirus expression system. The result should facilitate refined structural analysis and the development of new specific aldose reductase inhibitors for the treatment of diabetic complications.

Muscle fiber typing in routinely processed skeletal muscle with monoclonal antibodies

Muscle fiber typing is conventionally performed using mATPase enzyme histochemistry on cryostat sections. After pre-incubation of sections at pH 4.3, 4.6 and 10.3, based on the pattern of enzyme reactivity, the fibers can be classified in types I, II (subtypes A, AB and B) and the intermediate C (I and II) fibers. We have attempted to perform fiber typing of human psoas muscle by immunohistochemistry, using monoclonal antibodies R11D10 (specific for cardiac and type I skeletal myosin) and MY-32 (specific for fast muscle fibers) on cryostat as well as on paraffin sections. Staining of consecutive cryostat sections showed that type I fibers are R11D10 reactive whereas type II fibers are MY-32 reactive. Subtyping of type II fibers could not be performed by immunohistochemistry. Quantitative analysis of type I and II fibers showed that enzyme histochemical and immunohistochemical analysis are in close agreement.

Aldose reductase from human psoas muscle: Purification, substrate specificity, immunological characterization, and effect of drugs and inhibitors

Aldose reductase (ALR2) has been purified to homogeneity from human psoas muscle. From sodium dodecyl sulfate-polyacrylamide electrophoresis the enzyme is monomeric and has a molecular weight of 37,000. ALR2 catalyzes the primarily NADPH-dependent reduction of a wide variety of aldehydes, although the enzyme can also utilize NADH. The best substrates for ALR2 are aromatic aldehydes (e.g. pyridine-3-aldehyde; Km = 9 microM; kcat/Km = 150,000 s-1 M-1), while among aldoses DL-glyceraldehyde is the preferred substrate (Km = 72 microM; kcat/Km = 17,250). Low (100 microM) concentrations of CaCl2 and CaSO4 cause a marked inhibition (90%) of ALR2 as do higher concentrations (0.2 M) of MgCl2. (NH4)2SO4 caused a 2-fold activation of ALR2. The enzyme is also inhibited by quercetin and the commercially developed aldose reductase inhibitors alrestatin and sorbinil. ALR2 is inhibited only very slightly by sodium valproate and barbiturates. ALR2 cross-reacts immunologically with human brain and human placental aldose reductase and with ALR2 from monkey tissue. There is no precipitin cross-reaction of ALR2 with aldose reductases from other species nor with human aldehyde reductase 1 (ALR1) or with ALR1 from other species. The data show that human muscle is a new and relatively rich source of a monomeric NADPH/NADH reductase which is clearly identifiable as aldose reductase.

Aldose reductase from human psoas muscle: Affinity labeling of an active site lysine by pyridoxal 5′-phosphate and pyridoxal 5′-diphospho-5′-adenosine

The reaction of aldose reductase from human psoas muscle with either pyridoxal 5'-phosphate (PLP) or pyridoxal 5'-diphospho-5'-adenosine (PLP-AMP) results in a pseudo first-order 2-fold activation of the enzyme with the stoichiometric incorporation of 1 mol of either reagent per mol of enzyme. However, in addition to an increase in Vmax there was also an increase in Km for both substrate, DL-glyceraldehyde, and coenzyme, NADPH. This resulted in an overall decrease in catalytic efficiency (kcat/Km). Spectral analysis indicated that activation by both PLP and PLP-AMP was accompanied by Schiff's base formation and epsilon-pyridoxyllysine was identified in hydrolysates of the reduced enzyme PLP-complex. Digestion of either PLP-modified or PLP-AMP-modified aldose reductase with endoproteinase Lys-C followed by high performance liquid chromatography purification and amino acid sequencing of the pyridoxyllated peptide revealed that PLP and PLP-AMP had modified the same lysine residue. A 32-residue peptide containing the essential lysine was found to be highly homologous with a segment of the sequence of both human liver aldehyde reductase and rat lens aldose reductase. A tetrapeptide (Ile-Pro-Lys-Ser) containing the essential lysine was identical in all three enzymes. These results highlight the close structural similarity between members of the aldehyde reductase family.

Purification and characterization of human muscle pyruvate kinase

The M1 isozyme of pyruvate kinase has been purified from human psoas muscle in a seven-step procedure. Fractionation by ammonium sulfate precipitation, heat treatment, acetone precipitation, diethylaminoethyl cellulose batchwise treatment followed by chromatography on carboxymethyl cellulose and Sephadex G-200 gave a product with a specific activity of 383 U/mg representing a 294-fold purification with a yield of 11%. The product formed orthorhombic crystals and was homogeneous on polyacrylamide gel electrophoresis with and without sodium dodecyl sulfate, sedimentation velocity, sedimentation equilibrium, and immunodiffusion. The purified enzyme has a molecular weight of 240700 and has a sedimentation coefficient (S20,W) of 10.04S. It contains four subunits with identical molecular weights of 61000. No free N-terminal amino acids could be detected. Antibody prepared against the purified human M1 isozyme does not cross-react by immunodiffusion or enzyme inactivation with the human erythrocyte isozyme and in the reverse experiment antibody prepared against human erythrocyte pyruvate kinase does not cross-react with the purified M1 isozyme. The amino acid composition of the M1 isozyme is presented.