Percutaneous pinning of proximal humerus fractures: a biomechanical study

Mechanical testing of two-part surgical neck fractures fixed with four different pin configurations was performed. Ten fresh, frozen, unembalmed humeri stripped of all soft tissues were used; the surgical neck was osteotomized perpendicular to the humerus long axis. Terminally threaded 2.5-mm AO pins were used to fix the fracture. Humeri then were tested in both torsion and bending on a custom-made jig using Instron 1331 to assess the rigidity of pinning constructs. In torsion, two lateral pin construct was significantly less rigid than all other pin configurations. The addition of an anterior pin to two lateral pins did not increase bending rigidity, but significantly increased torsional stiffness. The addition of two bicortical tuberosity pins or two bicortical tuberosity pins and one anterior pin to two lateral pins significantly increased rotational and bending rigidity. Results confirm clinical data, and the authors conclude that multiplanar pins are needed to augment torsional stiffness, and that the addition of two bicortical tuberosity pins enhances bending rigidity.     

Controlled gene delivery by DNA-gelatin nanospheres

A novel system for gene delivery, based on the use of DNA-gelatin nanoparticles (nanospheres) formed by salt-induced complex coacervation of gelatin and plasmid DNA, has been developed. These particles were spherical, with a size range of 200-700 nm, contained 25-30% (w/w) DNA, and were stabilized by cross-linking of gelatin. As a consequence of being controlled by the cross-linking density of the gelatin matrix, the average release rate of DNA from nanospheres synthesized under standard conditions was 2.2%/day in serum. Nanosphere DNA incubated in bovine serum was more resistant to nuclease digestion than was naked DNA. Various bioactive agents could be encapsulated in the nanospheres by ionic interaction with the matrix components, physical entrapment, or covalent conjugation. Transfection of cultured cells with a luciferase plasmid was enhanced by conjugating human transferrin onto the nanosphere and coencapsulating the endolysolytic agent chloroquine. Under our experimental conditions, gene expression in mice subsequent to intramuscular injection of nanospheres containing 1 microg of a beta-galactosidase plasmid was greater and more prolonged than was observed after injection of an equal amount of naked DNA or DNA complexed with Lipofectamine.     

Research in gastrointestinal and abdominal radiology: barium and beyond

BACKGROUND: In The People's Republic of China in 1990, the age-specific seroprevalence of hepatitis A was investigated in eight large cities. METHODS: A stochastic model, the two-state Markov chain, was applied to hepatitis A virus seroprevalence data by age group. An age-specific risk rate, Markov Risk Rate (MRR), and its weighted sum, Total MRR, are defined and used as novel measure indices to prioritize age groups for allocating vaccine or to decide in which cities vaccine should be used to prevent hepatitis A. RESULTS: In 1990, the MRR1- in Xi'an, Jinan, Ha'erbin and Huhehaote, and the MRR10- in Chongqing were over 40. The MRR10- in Xi'an, Nanjing, Jinan and Ha'erbin and the MRR20- in Chongqing and Nanjing were over 20. The Total MRR in Chongqing and Ha'erbin were over 160, which was higher than the warning value. CONCLUSIONS: All age groups whose MRR was over 20 are strongly recommended to be vaccinated first. Chongqing and Ha'erbin are cities at high risk of a hepatitis A virus epidemic in the 1990s and therefore should be under close surveillance.     

Calcium current and GABA(B) receptors in dorsal sensory cells of the lamprey spinal cord

GABA and GABAB receptor agonists were shown to reduce the peak calcium current amplitude with its subsequent recovery, whereas glycine and taurine, the GABAA receptor agonists, did not modify the current. The findings suggest that the GABAB receptors mediate a presynaptic inhibition by suppression of the Calcium currents in the cyclostome spinal cord.     

Fatty acid carbon isotope ratios in humans on controlled diets

Carbon stable isotope ratios for six serum fatty acids (FA) are reported for human subjects on controlled fat diets to determine the range of natural isotope abundance and to demonstrate the leveling effect of a well-controlled diet. Twenty-nine subjects were randomly assigned to one of three controlled diets containing high, medium, or low fat. Diets were consumed for 8 wk. Serum samples were collected at baseline (0), 5, 6, 7, and 8 wk. FA were extracted and methylated. Isotope ratios were analyzed by high-precision gas chromatography combustion-isotope ratio mass spectrometry. At baseline, mean delta 13C for 16:0b, 16:1a, 18:0a, 18:1c,18:2n-6d and 20:4n-6bc were -24.1, -21.7, -21.6, -25.6, -29.6, and -25.0/1000, respectively, with an average standard deviation of 1.9/1000. Most delta 13C decreased during the diet period and appeared to have stabilized by week 5 at -25.3, -21.9, -22.3, -26.5, -30.1, and -24.5/1000, respectively. Between-subjected variability decreased from 1.74 to 1.20/1000 on the controlled diets. Measurement variability was 0.53/1000. The within-subject variability during weeks 5-8 was 0.57/1000 (range of 0.32-0.84/1000), showing a minimum biological fluctuation on controlled diets. There was no diet group effect on delta 13C of serum FA. Except for 18:2, the delta 13C of experimental diets was lower than that of serum FA, consistent with observations in animals. These data show that carbon isotope ratios stabilize in response to controlled diets within 5 wk, reflecting the isotope ratio of their dietary source, and establish isotope ratio fluctuations for endogenous compounds for future studies.     

Identification and characterization of glycosylation sites in human serum clusterin

Clusterin is a ubiquitous, heterodimeric glycoprotein with multiple possible functions that are likely influenced by glycosylation. Identification of oligosaccharide attachment sites and structural characterization of oligosaccharides in human serum clusterin has been performed by mass spectrometry and Edman degradation. Matrix-assisted laser desorption ionization mass spectrometry revealed two molecular weight species of holoclusterin (58,505 +/- 250 and 63,507 +/- 200). Mass spectrometry also revealed molecular heterogeneity associated with both the alpha and beta subunits of clusterin, consistent with the presence of multiple glycoforms. The data indicate that clusterin contains 17-27% carbohydrate by weight, the alpha subunit contains 0-30% carbohydrate and the beta subunit contains 27-30% carbohydrate. Liquid chromatography electrospray mass spectrometry with stepped collision energy scanning was used to selectively identify and preparatively fractionate tryptic glycopeptides. Edman sequence analysis was then used to confirm the identities of the glycopeptides and to define the attachment sites within each peptide. A total of six N-linked glycosylation sites were identified, three in the alpha subunit (alpha 64N, alpha 81N, alpha 123N) and three in the beta subunit (beta 64N, beta 127N, and beta 147N). Seven different possible types of oligosaccharide structures were identified by mass including: a monosialobiantennary structure, bisialobiantennary structures without or with one fucose, trisialotriantennary structures without or with one fucose, and possibly a trisialotriantennary structure with two fucose and/or a tetrasialotriantennary structure. Site beta 64N exhibited the least glycosylation diversity, with two detected types of oligosaccharides, and site beta 147N exhibited the greatest diversity, with five or six detected types of oligosaccharides. Overall, the most abundant glycoforms detected were bisialobiantennary without fucose and the least abundant were monosialobiantennary, trisialotriantennary with two fucose and/or tetrasialotriantennary. Clusterin peptides accounting for 99% of the primary structure were identified from analysis of the isolated alpha and beta subunits, including all Ser- and Thr-containing peptides. No evidence was found for the presence of O-linked or sulfated oligosaccharides. The results provide a molecular basis for developing a better understanding of clusterin structure-function relationships and the role clusterin glycosylation plays in physiological function.     

The helix-hinge-helix structural motif in human apolipoprotein A-I determined by NMR spectroscopy

The conformation of a synthetic peptide of 46 residues from apoA-I was investigated by fluorescence, CD, and 2D NMR spectroscopies in lipid-mimetic environments. ApoA-I(142-187) is mainly unstructured in water but helical in SDS or dodecylphosphocholine (DPC), although the peptide only associates with DPC at approximately the critical micellar concentration. Solution structures of apoA-I(142-187) were determined by distance geometry calculations based on 450 (in DPC-d38) or 397 (in SDS-d25) NOE-derived distance restraints, respectively. Backbone RMSDs for superimposing the two helical regions 146-162 and 168-182 are 0.98 +/- 0.22 (2.38 +/- 0.20) and 1.99 +/- 0.42 (2.02 +/- 0.21) A in DPC (SDS), respectively. No interhelical NOE was found, suggesting that helix-helix interactions between the two helical domains in apoA-I(142-187) are unlikely. Similar average, curved helix-hinge-helix structures were found in both SDS and DPC micelles with the hydrophobic residues occupying the concave face, indicating that hydrophobic interactions dominate. Intermolecular NOESY experiments, performed in the presence of 50% protonated SDS, confirm that the two amphipathic helices and Y166 in the hinge all interact with the micelle. The involvement of Y166 in lipid binding is supported by fluorescence spectroscopy as well. On the basis of all the data above, we propose a model for the peptide-lipid complexes wherein the curved amphipathic helix-hinge-helix structural motif straddles the micelle. The peptide-aided signal assignment achieved for apoA-I(122-187) (66mer) and apoA-I suggests that such a structural motif is retained in the longer peptide and most likely in the intact protein.     

Short-term exercise enhances insulin-stimulated GLUT-4 translocation and glucose transport in adipose cells

This investigation examined the effects of short-term exercise training on insulin-stimulated GLUT-4 glucose transporter translocation and glucose transport activity in rat adipose cells. Male Wistar rats were randomly assigned to a sedentary (Sed) or swim training group (Sw, 4 days; final 3 days: 2 x 3 h/day). Adipose cell size decreased significantly but minimally (approximately 20%), whereas total GLUT-4 increased by 30% in Sw vs. Sed rats. Basal 3-O-methyl-D-[14C]glucose transport was reduced by 62%, whereas maximally insulin-stimulated (MIS) glucose transport was increased by 36% in Sw vs. Sed rats. MIS cell surface GLUT-4 photolabeling was 44% higher in the Sw vs. Sed animals, similar to the increases observed in MIS glucose transport activity and total GLUT-4. These results suggest that increases in total GLUT-4 and GLUT-4 translocation to the cell surface contribute to the increase in MIS glucose transport with short-term exercise training. In addition, the results suggest that the exercise training-induced adaptations in glucose transport occur more rapidly than previously thought and with minimal changes in adipose cell size.     

Signalling functions of protein palmitoylation

Covalent lipid modifications anchor numerous signalling proteins to the cytoplasmic face of the plasma membrane. These modifications mediate protein-membrane and protein-protein interactions and are often essential for function. Protein palmitoylation, due to its reversible nature, may be particularly important for modulating protein function during cycles of activation and deactivation. Despite intense investigation, the exact functions of protein palmitoylation are not well understood. However, it is clear that palmitoylation can affect a protein's affinity for membranes, subcellular localization, and interactions with other proteins. In this review, recent advances in understanding the functions and mechanisms of protein palmitoylation are discussed, with particular emphasis on how this lipid affects the biochemistry and cell biology of signalling proteins.     

Enhancement of antipsychoticlike properties of raclopride in rats using the selective serotonin2A receptor antagonist MDL 100,907

OBJECTIVE: To study the superior-inferior stabilizing functions of the coracohumeral ligament (CHL) and the rotator interval capsule (RIC) with use of a material testing machine. MATERIAL AND METHODS: The axial translations of the humerus with the superior-inferior translation force of 30 N applied were recorded under the following joint capsule conditions: (1) intact, (2) vented, (3) the CHL sectioned, and (4) the RIC incised in six cadaver shoulders. The order of sectioning was changed for conditions 3 and 4 in six other cadaver shoulders. RESULTS: With the arm in internal and neutral rotations, venting the capsule significantly increased the superior-inferior translation, which was unaffected by further sectioning of the CHL and the RIC. With the arm in external rotation, only the CHL contributed significantly to inferior stability, whereas both this ligament and the RIC contributed to superior stability to a lesser degree. CONCLUSION: The CHL is a stabilizer in superior inferior directions with the arm in external rotation, and the intra-articular pressure that is maintained by the intact RIC is a stabilizer in superior-inferior directions with the arm in internal and neutral rotations. These findings may provide a scientific background to support closure of the interval space to stabilize the shoulder and may explain part of the superior instability observed in shoulders with rotator cuff tears.