Anatomical variations of the biceps brachii insertion: a proposal for a new classification.

BACKGROUND: The biceps brachii (BB) muscle is one of the three muscles located in the anterior compartment of the arm. Its insertion consists of two parts. The first part - main tendon - attached in the radial tuberosity and the second part - lacertus fibrosus (LF) - in the fascia of the forearm flexors. The intention of research was to reveal the morphological diversity of the insertion of this muscle. Thanks to the results of this work, have been created a classification of the distal attachment of BB. The results of that research can be used to further develop surgical procedures in the given region. MATERIALS AND METHODS: Eighty (40 left, and 40 right, 42 female, 38 male) upper limbs fixed in 10% formalin solution were examined. RESULTS: We observed three types of the insertion of the BB. Type I was characterised by a single tendon and occurred most frequently in 78.75% of the examined limbs. The second most common type was type II which was characterised by a double tendon and was observed in 13.75% of all the limbs. The last and least common was type III which was characterised by three tendons and occurred in 7.5% of the examined limbs. Additionally, the type of LF was analysed. In 8 (10%) specimens it was absent, i.e. in 2 specimens with type II insertion and 6 specimens with type III (p = 0.0001). Therefore, it may be deduced that type III BB insertion tendon predisposes to LF deficiency. CONCLUSIONS: The BB tendon is characterised by high morphological variability. The new classification proposes three types of distal attachment: type I - one tendon; type II - two separated band-shaped tendons; type III - three separated band-shaped tendons. The presence of type III BB tendon predisposes to a lack of LF.

A guideline to medical photography: a perspective on digital photography in an orthopaedic setting.

PURPOSE: Quality photographs are essential for clinical documentation, research, and publication in scientific journals and teaching. Oftentimes, non-ideal lighting and a sterile environment restrict the medical photographer, resulting in lower-quality photographs. This article aims to provide a clear and comprehensible guideline for medical photography in an orthopaedic setting. METHODS: This article is based on extensive photographic involvement in operating and laboratory settings, in close collaboration with medical professionals from the Steadman Clinic (Vail, Colorado, USA), Gothenburg University (Göteborg, Sweden) and Erasmus MC (Rotterdam, the Netherlands). Background literature was searched through Google Scholar and PubMed. RESULTS: Three relevant journal articles, and one book on medical photography, were used to write this paper. Seventeen Internet articles were used for background information. CONCLUSION: A relevant, up-to-date and comprehensive guideline to medical photography for medical professionals, with or without photographic experience, is provided. LEVEL OF EVIDENCE: Expert opinion, Level V.

Properties of the porin of Haemophilus influenzae type b in planar lipid bilayer membranes.

The major outer membrane protein (40 kDa) of the bacterium Haemophilus influenzae type b is a porin which forms transmembrane permeability channels. It has an exclusion limit for oligosaccharides of about 1.4 kDa. When this protein was added to the aqueous phase which was bathing a planar lipid bilayer, it caused the conductance of the membrane to increase by several orders of magnitude. At low protein concentrations (2-10 pM), the conductance of the membrane increased in a stepwise fashion with an average single-channel conductance of 1.1 nS in 1 M KCl. Single-channel experiments were performed with a variety of different salts. The conductance of single channels was proportional to the specific conductance of the aqueous solution which was bathing the membrane. Current through the pores was proportional to the applied voltage, indicating that these pores are not voltage-controlled. The 40 kDa porin was very slightly cation-selective: the pores were about 1.6-times more permeable to potassium ions than to chloride ions. These properties of the 40 kDa porin are those of large water-filled channels and are characteristic of most bacterial porins. The single-channel conductance of the porin is, however, much smaller than might be expected from its exclusion limit. A model is proposed which could explain the differences in apparent pore size.

Unfolding affects insect cell permeabilization by Bacillus thuringiensis Cry1C toxin.

Bacillus thuringiensis Cry toxins are efficient, environment-friendly biological insecticides. Their molecular mode of action on target insect cells remains largely unknown. The aim of this study was to investigate the relation between the conformational state of the Cry1C toxin and its ionophoric activity on live Sf9 cells of Spodoptera frugiperda, a target insect for this protein. Potassium ion movement induced by Cry1C across the cell membrane was measured with a fluorescent assay developed previously and the conformation of the toxin was studied using tryptophan spectroscopy. Following treatment with 4 M guanidinium hydrochloride, which resulted in the unfolding of its N-terminal half, the toxin retained its full capacity to permeabilize the cells while the fully unfolded toxin did not induce potassium leakage. Therefore, permeabilization of Sf9 cells by Cry1C requires the integrity of the C-terminal half of the toxin and may depend on an initial unfolding step provided by the acidic environment of the cells.

Mechanism of neomycin stimulation of D-glucose uptake in rabbit intestinal brush border membrane.

In order to study the effect of the antibiotic neomycin on the intestinal epithelium, D-glucose was used as a probe molecule and its transport into rabbit brush border membrane vesicles was measured by a rapid filtration method. Treatment of the epithelium with neomycin sulfate prior to the preparation of the brush border membrane enhanced the D-glucose uptake, whereas neutral N-acetylated neomycin did not. This action of neomycin was related to its polycationic character and not to its bactericidal action. No significant difference could be demonstrated between the protein content or disaccharidase-specific activities of the brush border fractions from treated or non-treated intestines. Electrophoretic protein patterns of SDS-solubilized membrane were not significantly different after neomycin treatment. To gain more information on the mechanism involved in the stimulation of D-glucose transport, experiments were conducted on phosphatidyl glycerol artificial membranes and the results compared with those obtained with brush border membrane. At a concentration of 10(-7) M, neomycin decreased the nonactin-induced K+ conductance by a factor of approx. 100. The membrane conductance was linearly dependent on the neomycin concentration and the conductance in 10(-2) M KCl was 10 times that in 10(-3) M KCl. The valence of neomycin was estimated, from the slope of these curves, to be between 6 and 4. In contrast, acetylated neomycin had no effect on the nonactin-induced K+ membrane conductance. Therefore, the effect of neomycin on artificial membrane is related to its 4 to 6 positive charges. It is proposed that the stimulation of sugar transport in brush border membrane is related to screening of the membrane negative charges by the positively-charged neomycin. Accumulation of anions at the membrane surface then occurs and their diffusion into the intravesicular space would increase the transmembrane potential which, in turn, stimulates the entry of D-glucose.

Voltage gating of porins from Haemophilus influenzae type b.

The major outer membrane protein of Haemophilus influenzae type b (Hib) is porin (M(r) 37,782; 341 amino acids). Porins were purified from Hib strains representative of the three outer membrane protein subtypes 1H, 2L and 6U, reconstituted into artificial planar bilayers, and tested for their voltage dependency. At membrane potentials of 50-80 mV, individual Hib 2L and 6U porin channels showed a high probability of undergoing a reversible change to one of several lower conducting substates. Such behaviour was not observed for Hib 1H porin with transmembrane potentials up to 80 mV. The voltage dependence of Hib 2L and 6U porins was asymmetric: it occurred at only one polarity. The asymmetry was also observed for membranes with numerous porins incorporated, suggesting that Hib porin inserted asymmetrically into the bilayer. At macroscopic levels the voltage gating reduced the conductance by 25-50%, implying that the channels closed only partially. Hib 2L porin differs from Hib 1H porin by the substitution Arg166Gln and Hib 6U porin differs from Hib 1H porin by substitutions at ten amino acids including the change Arg166Leu. We conclude that substitutions at Arg166 residue, which is localized to surface-exposed loop number four, are associated with a lowered threshold potential for the voltage gating of Hib porin. This surface-exposed loop may play some role in the conformational changes that occur during voltage gating.

Activation of an ATP-dependent K(+) conductance in Xenopus oocytes by expression of adenylate kinase cloned from renal proximal tubules.

In rabbit proximal convoluted tubules, an ATP-sensitive K(+) (K(ATP)) channel has been shown to be involved in membrane cross-talk, i.e. the coupling (most likely mediated through intracellular ATP) between transepithelial Na(+) transport and basolateral K(+) conductance. This K(+) conductance is inhibited by taurine. We sought to isolate this K(+) channel by expression cloning in Xenopus oocytes. Injection of renal cortex mRNA into oocytes induced a K(+) conductance, largely inhibited by extracellular Ba(2+) and intracellular taurine. Using this functional test, we isolated from our proximal tubule cDNA library a unique clone, which induced a large K(+) current which was Ba(2+)-, taurine- and glibenclamide-sensitive. Surprisingly, this clone is not a K(+) channel but an adenylate kinase protein (AK3), known to convert NTP+AMP into NDP+ADP (N could be G, I or A). AK3 expression resulted in a large ATP decrease and activation of the whole-cell currents including a previously unknown, endogenous K(+) current. To verify whether ATP decrease was responsible for the current activation, we demonstrated that inhibition of glycolysis greatly reduces oocyte ATP levels and increases an inwardly rectifying K(+) current. The possible involvement of AK in the K(ATP) channel's regulation provides a means of explaining their observed activity in cytosolic environments characterized by high ATP concentrations.

Video imaging analysis of the plasma membrane permeabilizing effects of Bacillus thuringiensis insecticidal toxins in Sf9 cells.

The size and ionic selectivity of the pores formed by the insecticidal crystal protein Cry1C from Bacillus thuringiensis in the plasma membrane of Sf9 cells, an established cell line derived from the fall armyworm Spodoptera frugiperda, were analyzed with a video imaging technique. Changes in the permeability of the membrane were estimated from the rate of osmotic swelling of the cells. In the presence of Cry1C, which is toxic to Sf9 cells, the permeability of the cell membrane to KCl and glucose increased in a dose-dependent manner. In contrast, Cry1Aa, Cry1Ab and Cry1Ac, toxins to which Sf9 cells are not susceptible, had no detectable effect. Pores formed by Cry1C allowed the diffusion of sucrose, but were impermeable to the trisaccharide raffinose. On the basis of the hydrodynamic radii of these substances, the diameter of the pores was estimated to be 1.0-1.2 nm. In the presence of salts, the rate of swelling of cells exposed to Cry1C was about equally influenced by the size of the anion as by that of the cation, indicating that the ionic selectivity of the pores is low.

Temperature dependence of endocytosis in renal epithelial cells in culture.

Temperature dependence of fluid-phase endocytosis was determined in two renal epithelial cell lines, MDCK cells and LLC-PK1 cells, using Lucifer Yellow or horseradish peroxidase as markers. For both cell lines, grown on solid support as a confluent monolayer, biphasic curves of marker uptake vs. temperature were obtained. The changes in slope occurred around 27 degrees C, a critical temperature at which the lipids of the plasma membrane of MDCK cells enter in the gel state. Activation energies were significantly higher above 27 degrees C (15-22 kcal/mol) than below that critical temperature (9-12 kcal/mol). These data indicate that changes in membrane physical state have marked effects on endocytic processes. They suggest that two mechanisms, with different activation energies are involved in the fluid phase endocytosis by renal epithelial cells in culture.

Ionic channels induced by surfactin in planar lipid bilayer membranes.

Surfactin is a lipopeptide produced by certain strains of Bacillus subtilis and has potent surface activity. Here, we present the first results showing that ion-conducting pores can be formed by surfactin in artificial lipid membranes. With a low aqueous concentration of surfactin (1 microM) and a restricted membrane area (5.10(-5) cm2) we observed conductance jumps that indicate the formation of individual ionic channels in the presence of K+, Rb+, Cs+, Na+ or Li+ chlorides. Although for every salt concentration (Ci), the distribution in amplitude of the conductance steps (lambda i) may be rather broad, there is always a step amplitude which is more frequent than the others. In addition, the channels corresponding to this most frequent step amplitude are the longest in duration. For Ci = 1 M, the cationic selectivity sequence deduced from these most frequent events is K+ greater than Rb+ greater than Na+ greater than Cs+ = Li+ with respective values for lambda Mi: 130, 110, 80 and 30 pS. In KCl solutions lambda MKCl increases as a function of Ci for low Ci, and shows a plateau for Ci greater than 0.5 M. When measured on larger area membranes (10(-2)cm2) with 1 M solutions of the monovalent salts KCl, NaCl, RbCl and CsCl or the divalent salt CaCl2, the macroscopic low voltage conductance (G0) increases with a slope of 2 on a log-log plot as a function of surfactin concentration. These results demonstrate that surfactin produces selective cationic channels in lipid bilayer membranes and suggest that at higher salt concentration, a dimer is involved in this functional channel-forming process.

Molecular size of the renal sodium/phosphate symporter in native and reconstituted systems.

The size of the renal sodium/phosphate symporter was estimated with the radiation inactivation technique in isolated bovine brush border membrane vesicles and after reconstitution in proteoliposomes. The functional unit of the native phosphate carrier had a radiation inactivation size of 172 +/- 17 kDa. Identical values were obtained for the reconstituted carrier whether it was irradiated before or after the formation of the proteoliposomes (161 +/- 9 and 159 +/- 11 kDa, respectively). The sodium-independent uptake of phosphate was not affected significantly by radiation doses up to 10 Mrad. This activity is therefore not due to the reconstitution of a large phosphate-binding protein such as alkaline phosphatase. Furthermore, bromotetramisole, a specific inhibitor of phosphate binding to this enzyme, had no significant effect on the uptake of phosphate by the proteoliposomes.

Factors affecting the stability of the renal sodium/phosphate symporter during its solubilization and reconstitution.

Phosphate is reabsorbed across the brush-border membrane of the proximal tubule by a specific sodium-dependent symporter. Like the other brush-border membrane transport proteins of the kidney, the phosphate carrier remains to be isolated in a functional state. To establish a set of parameters that allow to preserve its biological activity, the phosphate carrier was solubilized under systematically varied conditions and reconstituted into proteoliposomes. Successful reconstitution was achieved only when the extraction buffer contained lipids extracted from the renal brush-border membrane. Glycerol, an osmolyte which reduces the water activity of the solution, was also required. It could however be replaced by 150 mM sodium or potassium phosphate. Below this concentration and in the presence of glycerol, the ionic strength of the solution had little effect on the stability of the transporter, but sodium phosphate could not be replaced by sodium chloride. Phosphate transport in reconstituted vesicles depended on the concentration of detergent and pH of the extraction buffer. Finally, transport activity was increased when solubilization was carried out in the presence of a reducing agent, dithiothreitol. These results should be helpful during the purification and further characterization of the renal phosphate symporter.

Purification and refolding of recombinant Haemophilus influenzae type b porin produced in Bacillus subtilis.

The major diffusion channel in the outer membrane of Haemophilus influenzae type b (Hib) is porin (341 amino acids; Mr 37 782). The Hib porin gene was cloned and overexpressed in Bacillus subtilis. Recombinant Hib porin (Bac porin), having aggregated into inclusion bodies, was purified under denaturing conditions and subsequently refolded. To compare Bac porin that is intrinsically devoid of lipooligosaccharides versus native Hib porin, the properties of Bac porin were assessed by the following four criteria: circular dichroism spectroscopy, channel formation in planar bilayers, resistance to trypsin digestion and formation of the conformational epitope recognized by an anti-Hib porin monoclonal antibody. We conclude that in the absence of lipooligosaccharides, Bac porin was refolded into a functional form which closely resembled the structure of Hib porin.

Ion channels formed in planar lipid bilayers by Bacillus thuringiensis toxins in the presence of Manduca sexta midgut receptors.

A purified, GPI-linked receptor complex isolated from Manduca sexta midgut epithelial cells was reconstituted in planar lipid bilayers. CryIAa, CryIAc and CryIC, three Bacillus thuringiensis insecticidal proteins, formed channels at much lower doses (0.33-1.7 nM) than in receptor-free membranes. The non-toxic protein CryIB also formed channels, but at doses exceeding 80 nM. The channels of CrylAc, the most potent toxin against M. sexta, rectified the passage of cations. All other toxin channels displayed linear current-voltage relationships. Therefore, reconstituted Cry receptors catalyzed channel formation in phospholipid membranes and, in two cases, were involved in altering their biophysical properties.

Cry1Ac, a bacillus thuringiensis toxin, triggers extracellular Ca2+ influx and Ca2+ release from intracellular stores in Cf1 cells

Intracellular Ca2+ concentration was measured in single Cf1 cells (Choristoneura fumiferana, spruce budworm) loaded with Fura-2, a Ca2+-sensitive fluorescent probe. Cf1 cells displayed Ca2+ surges in response to Cry1Ac and Cry1C proteins, two Cf1-toxic Bacillus thuringiensis products, but not to Cry1Aa and Cry3A, which are not toxic to Cf1 cells. In the presence of extracellular Ca2+, the toxin-induced Ca2+ response was insensitive to methoxyverapamil, a voltage-dependent Ca2+ channel blocker, but was abolished by lanthanum, a general inhibitor of Ca2+ transport. In the absence of external Ca2+, Cry1Ac induced a small intracellular Ca2+ transient which was inhibited by TMB-8, a blocker of Ca2+ release from inositol-1,4,5-trisphosphate-sensitive pools. Under these conditions, thapsigargin, which inhibits intracellular Ca2+-ATPases, elicited a Ca2+ surge when applied alone. However, subsequent addition of Cry1Ac failed to induce a Ca2+ signal, indicating a depletion of intracellular Ca2+ pools. In Cf1 cells, therefore, bioactive B. thuringiensis toxins triggered intracellular Ca2+ surges which were mainly due to the influx of extracellular Ca2+ through toxin-made pores, as confirmed by planar lipid bilayer experiments. Furthermore, TMB-8- and thapsigargin-sensitive Ca2+ stores contributed to the Cry1Ac-induced Ca2+ signal.

Injury surveillance at the USTA Boys' Tennis Championships: a 6-yr study.

Injuries that required physical or medical assistance were recorded for participants at the United States Tennis Association National Boys' Tennis Championships from 1986-1988, 1990-1992 (N = 1440, 240 athletes per year). Over the 6-yr period, a total of 304 athletes (or 21.1%) sustained new or recurrent injuries that required evaluation by the medical team. New injuries alone numbered 145 (incidence rate of 9.9 per 100 athletes). The analysis of injuries showed a higher rate of lower than upper extremity injuries. When evaluated by anatomic regions, back injuries were most common followed by thigh, shoulder, and ankle injuries, respectively. When evaluated by injury type, strains and sprains were most common (71% of all injuries) with fractures and dislocations being rare (1.3% of all injuries). The lower extremity provided the majority of sprain type injuries with 87.5% of ligament sprains coming from the knee and ankle. Injuries with tennis eponyms (i.e., tennis toe, tennis leg, tennis elbow, and tennis shoulder) were rare (0%-5% of all injuries). It would appear that these young elite athletes are at significant risk of injury.

The anatomy of the deep infrapatellar bursa of the knee.

Disorders of the deep infrapatellar bursa are important to include in the differential diagnosis of anterior knee pain. Knowledge regarding its anatomic location can aid the clinician in establishing a proper diagnosis. Fifty cadaveric knees were dissected, and the deep infrapatellar bursa had a consistent anatomic location in all specimens. The deep infrapatellar bursa was located directly posterior to the distal 38% of the patellar tendon, just proximal to its insertion on the tibial tubercle. There was no communication to the knee joint. Its average width at the most proximal margin of the tibial tubercle was slightly wider than the average distal width of the patellar tendon. It was found to be partially compartmentalized, with a fat pad apron extending down from the retropatellar fat pad to partially divide it into anterior and posterior compartments. The recommended approach to this bursa, either for aspiration, injection, or surgery, is along the lateral edge of the patellar tendon just proximal to the tibial tubercle.

Arthroscopic evaluation of the lateral compartment of knees with grade 3 posterolateral knee complex injuries.

The purpose of this study was to evaluate prospectively the arthroscopic findings in the lateral compartment of knees with posterolateral knee complex injuries, to help identify individual injured anatomic structures, and to assist in the clinical identification of these injuries. Thirty of 33 consecutive knees noted to have grade 3 posterolateral knee complex injuries on preoperative evaluations and examinations under anesthesia underwent arthroscopic evaluation concurrently with open reconstruction. The arthroscopic evaluation revealed a significant number of pathologic changes in the lateral compartment that may have gone undetected if only an open reconstruction had been performed. Tears were identified arthroscopically in 25 (83%) of the anteroinferior, 22 (73%) of the posterosuperior, and 14 (47%) of the posteroinferior popliteomeniscal fascicles. Injuries to the coronary ligament (80%) and meniscotibial portion of the midthird lateral capsular ligament (73%) were also frequently seen. Other structures injured included the meniscofemoral portion of the posterior capsule (37%), the ligament of Wrisberg (33%), and the meniscofemoral portion of the midthird lateral capsular ligament (10%). Ten avulsions (33%) of the popliteal tendon origin off the femur were also identified. Identification of these injured components greatly facilitated open reconstruction of injuries to the posterolateral complex of the knee. All 30 knees were noted to have greater than 1 cm of lateral joint laxity with application of a varus stress. When an unexpected amount of lateral joint laxity is seen arthroscopically (a "drive-through" sign) in a patient with suspected ligamentous instability, one should consider a diagnosis of posterolateral knee complex injury.