Microbial production of natural δ-decalactone and δ-dodecalactone from the corresponding α,β-unsaturated lactones in Massoi bark oil

Summary The hydrogenation of ,-unsaturated Massoi lactones to natural -lactones by various microorganisms belonging to the Basidiomycetes and by Saccharomyces ce cerevisiae has been studied. Natural -decalactone is an important constituent of several natural flavourings. Process parameters for the hydrogenation by baker's yeast have been characterized on a 2-1 fermentation scale. High hydrogenation activity by baker's yeast was observed at pH 5.5, a temperature of 35° C, no oxygen limitation and constant addition of glucose. By stepwise addition of 2-decen-5-olide about 1.2 g/l of 5-decanolide could be obtained in a fermentation of 16 h. The same concentration could be obtained in 8 h by adding all the substrate at once (1.7 g/l) in the presence of 2% cyclodextrin.Offprint requests to: P. H. van der Schaft     

Separation of the pineal vesicle from the wall of the third ventricle during the post-hatching development of the chicken

Summary According to light- and electron-microscopic observations the pineal organ of the 3-day-old chicken consists of a prominent end vesicle and a tapering parenchymal stalk. During this stage the pineal lumen is in open communication with the third ventricle. However, in the 40-day-old chicken, which still possesses a well-developed end vesicle, the proximal portion of the pineal stalk displays regressive changes leading to local fragmentation. At this stage the pineal stalk is reduced, and the pineal lumen is missing. In 1-year-old chickens the parenchyma of the proximal portion of the stalk is further diminished, and in 3-year-old domestic fowl is completely displaced by bundles of collagenous fibers, only some nerve fibers being present. This post-hatching pineal development may reflect the sequence of changes leading from pineal sense organs to pineal glands.This work was supported by a grant-in-aid for Scientific Research from the Ministry of Education, Science and Culture of Japan     

Anatomic contribution to the study on the variety of fibrous formations of the arm in humans

The authors have examined an anatomical preparation of a human upper limb preserved in the Anatomical Museum in Bologna. The specimen was formed by bone, fibrous formations, musculus biceps brachii and musculus coracobrachialis. The soft parts were kept in situ and dried by mummification. The humerus showed abnormalities at its proximal extremity (Fig. 1) and the muscles displayed fibrous varieties: a) a fibrous sheet (Fig. 1, 2) connected the caput brevis of the musculus biceps brachii to the articular capsule of the shoulder joint; b) an aponevrosis (Fig. 1, 3) connected the musculus coracobrachialis to the same articular capsule and the humerus. These observations were discussed from an evolutionary and functional point of view.     

The respective roles of plastic and orthopedic surgery in limb salvage

The evolution of techniques in plastic surgery and orthopedic surgery over the past few decades has enabled a great level of success in limb salvage. Limb salvage can now be achieved when faced with trauma, tumor, sepsis, or vascular disease. In fact, "What can be salvaged?" is now a less common debate among clinicians than "What should be salvaged?" Often discussions among surgeons from various subspecialties, including orthopedics, plastics, trauma, and vascular surgery, are characterized by how each of them can perform their respective part of the salvage operation, be it bony fixation, revascularization, or soft-tissue coverage, but none of them is certain whether it should be attempted. What is needed in these clinical situations is an interdisciplinary team approach led by individual or groups of clinicians who are familiar not only with their own subspecialized skills but also with those of their colleagues and the outcomes associated with integrated efforts at limb salvage. The concept of orthoplastic surgery is based on such an idea, where the combined skills and techniques of the orthopedic surgeon and reconstructive microsurgeon are used in concert to direct efforts toward limb salvage or decide against it when it is not indicated. This article presents a review of the roles of the two subspecialties and how an orthoplastic team can function with the current techniques to improve outcomes in limb salvage surgery.     

T-cell repertoire in a strain of transgenic C57BL/6 mice with the HLA-DRA gene on the X-chromosome

We have established a strain of transgenic mice in which the HLA-DRA gene was integrated into the X-chromosome and the xenogeneic mixed isotype molecule, DRE^b, was expressed in a cell type-specific manner, although the transgenic DRA gene contained only 268 base pairs of the 5-flanking region. The DRE^b molecules expressed in the transgenic mice functioned as major histocompatibility complex (MHC) class II to select T-cell repertoire, and to stimulate mixed lymphocyte reaction. In female transgenic mice homozygous for HLA-DRA (DR-B6-F-homo) and male transgenic mice (DR-B6-M), DRE^b molecules were expressed in almost all of the MHC class II A^b-positive cells. In contrast, the expression of DRE^b molecules in female transgenic mice hemizygous for HLA-DRA (DR-B6-F-hemi) was found only in part of the A^b positive cells, and the proportion of cells expressing the DRE^b molecules varied due to random inactivation of one of the X-chromosomes. Clonal deletions of the T cells and mature thymocytes bearing Tcrb-V5 and Tcrb-V11, which are eliminated from the peripheral repertoire in mice expressing self-superantigen and MHC class II E molecules, were incomplete in DR-B6-F-hemi as compared with those in DR-B6-F-homo, and were correlated with the proportion of DRE^b-positive spleen cells. These observations suggested that the number of bone marrow-derived cells expressing DRE^b molecules was critical for clonal deletions of Tcrb-V5⁺ and Tcrb-V11⁺ T cells in the thymus.     

The role of the zone of polarizing activity in controlling the differentiation of the apical mesenchyme of the chick wing-bud: histochemical thechniques in the analysis of a developmental problem

Summary Following excision of the posterior half of the three-day chick wing bud, the anterior half which normally forms humerus (part), radius and digit 2, forms only a single skeletal element (humerus, of humerus fused with reduced radius). If part of the zone of polarizing activity is included in the remaining anterior part of the wing bud, a normal wing with normal skeleton forms. Excision of the anterior half of the chick wing-bud results in the posterior half forming humerus (part), ulna and digits 3–5. This is confirmed as the normal prospective fate of the posterior half by chimeric quail-chick wing-buds in which Feulgen staining of the nucleolus-associated heterochromatin of quail cells enables their contribution to the resultant skeleton to be identified. Beginning at 18 h alter posterior half amputation, the anterior distal mesenchyme becomes necrotic and the apical e todernal ridge regresses. By contrast, following anterior half amputation, posterior halves develop no more cell death than control wing-buds.Anterior half regression is characterized by cell fragmentation and phagocytosis. First, in both the apical ectodermal ridge and distal mesenchyme cells, acid phosphatase-rich autophagic bodies appear, the cells then becoming autolytic (with diffuse acid phosphatase activity) and fragmenting. Neighbouring cells phagocytose the dead cell fragments, the mesenchyme cells forming large non-professional macrophages containing many acid phosphatase-rich vacuoles.These experiments show that for survival and differentiation, the anterior and distal mesenchyme of the wing bud requires a factor from the posterior part, thus suggesting that the zone of polarizing activity controlsantero-posterior differentiation in the normal wing.     

No evidence for linkage between the loci for coagulation factor XIII-A and HLA

Summary The linkage analysis between the locus for coagulation factor XIII-A (F13A) and HLA region genes (HLA-A,-C,-B) was performed. In males, the maximum of lod scores between F13A and HLA was 0.33 at =0.30, and in females lod scores were negative at all values of . The results provided no evidence for close linkage between F13A and HLA genes.     

Normalization of electromyogram in the neck-shoulder region

Linear and curvilinear electromyogram (EMG) normalization methods were compared among ten healthy men during a simulated work cycle demanding attention and static holding of the arm (Solitaire test). Maximal voluntary contractions (MVC) and gradually increasing contractions up to 70% of MVC were used for normalization in different arm postures. The test contractions studied included inward and outward rotations, abduction, shoulder elevation, and flexion in different arm positions. The shoulder load moment was calculated for the flexion tests using a simple two-dimensional model. The effect of arm posture on the EMG versus shoulder load moment relationship was studied on the following muscles: supraspinatus, infraspinatus, trapezius (three parts), deltoid (two parts) and pectoralis major. All muscles participated in the MVC tests performed, and its was not possible to suggest a single recommended test for each muscle. Differences in normalized EMG median values ranging up to 30% of MVC were found between linear and curvilinear normalization methods. Short-term repeatability of normalization based on a contraction with gradually increasing force was good. Arm posture affected the relationships between shoulder load moment and EMG activity of all muscles studied. Arm posture did not, however, have a significant effect on the estimated amplitude probability distribution functions during the simulated work task. Therefore, at least for the tasks studied, the principle of normalizing in the middle position of the range of movement was deemed acceptable.     

Development and evaluation of the measurement system for the human shoulder joint based on the 6 DOF kinematic modelling

Although numerical models on the shoulder complex joint are currently available, many are impractical because of the procedural complexity coupled with limited and mere simple simulations. The present study defined the clavicle-scapula system as the "base of the humerus" in determining the position of proximal head of humerus, rendering conclusive innovation of a six degree of freedom (DOF) shoulder complex joint model. Furthermore, a complete measurement system where evaluation by calibrating the actual values via the use of an electromagnetic tracking device (ETD) was developed based on the innovated model. The special calibration method using optimizing calculation to work out the rotational center of humerus was employed and actually tested if the theoretical consideration was practically available. As a result of accuracy check experiments, the measurement error was defined within 2-3 mm, indicating sufficient accuracy in studies for human movement. Our findings strongly advocate that the benefit of this novel measurement system would contribute to studies related to shoulder movements in physiological anthropology.     

Evidence for sequential appearance of cartilage matrix proteins in developing mouse limbs and in cultures of mouse mesenchymal cells

The initiation of synthesis and the accumulation of four cartilage matrix proteins (type II collagen and three noncollagenous proteins, one of Mr 148, one of Mr 59, and an oligometric protein of Mr above 500 with 100-kDa subunits, respectively) were studied in developing mouse limbs and in cultures of limb bud mesenchyme by means of immunolocalization. On day 13 of gestation, type II collagen was observed throughout the entire humerus, whereas the 148-kDa protein was localized only in the central portion. Neither the 100-kDa-subunit protein nor the 59-kDa protein could be demonstrated in the humerus at that stage. On day 14 1/2, type II collagen and the 148-kDa protein were codistributed throughout the humerus. The 100-kDa-subunit protein was detectable in the periphery of the humerus, whereas little 59-kDa protein could yet be demonstrated. On day 18, all four proteins being studied could be detected immunologically in the developing mouse humerus. They differed in immunolocalization. Type Ii collagen, the 148-kDa protein, and the 100-kDa-subunit protein were codistributed throughout the distal and proximal parts of the cartilage. However, the 148-kDa protein could no longer be detected immunochemically in the outermost part of the cartilage in the proximal shoulder joint. The 148-kDa protein codistributed with type II collagen and the 100-kDa-subunit protein in the distal cartilaginous region, where joint development was less advanced. On the other hand, the 59-kDa protein was not demonstrated directly within the hyaline cartilaginous structures, but surrounded the entire structure. This protein was also present in the same part of the proximal joint region as that in which the 148-kDa protein was not detected. To develop an in vitro model for studies of skeletogenesis, mesenchymal cells prepared from mouse limb buds were cultured as micromass cultures at high initial cell density to favor chondrogenesis. On day 3 of culture, type II collagen was the only protein that could be detected immunochemically in the cultures, whereas on day 6 the 148-kDa protein was demonstrated, and a few chondrocytes in the central portion of each cartilaginous nodule were associated with the 100-kDa-subunit protein. The 59-kDa protein could not yet be immunochemically detected.(ABSTRACT TRUNCATED AT 400 WORDS)     

The Mechanical Benefit of Medial Support Screws in Locking Plating of Proximal Humerus Fractures

Background

The purpose of this study was to evaluate the biomechanical advantages of medial support screws (MSSs) in the locking proximal humeral plate for treating proximal humerus fractures.

Methods

Thirty synthetic left humeri were randomly divided into 3 subgroups to establish two-part surgical neck fracture models of proximal humerus. All fractures were fixed with a locking proximal humerus plate. Group A was fixed with medial cortical support and no MSSs; Group B was fixed with 3 MSSs but without medial cortical support; Group C was fixed with neither medial cortical support nor MSSs. Axial compression, torsional stiffness, shear stiffness, and failure tests were performed.

Results

Constructs with medial support from cortical bone showed statistically higher axial and shear stiffness than other subgroups examined (P<0.0001). When the proximal humerus was not supported by medial cortical bone, locking plating with medial support screws exhibited higher axial and torsional stiffness than locking plating without medial support screws (P≤0.0207). Specimens with medial cortical bone failed primarily by fracture of the humeral shaft or humeral head. Specimens without medial cortical bone support failed primarily by significant plate bending at the fracture site followed by humeral head collapse or humeral head fracture.

Conclusions

Anatomic reduction with medial cortical support was the stiffest construct after a simulated two-part fracture. Significant biomechanical benefits of MSSs in locking plating of proximal humerus fractures were identified. The reconstruction of the medial column support for proximal humerus fractures helps to enhance mechanical stability of the humeral head and prevent implant failure.     

Histochemical detection of sugar residues in the chick embryo mesonephros with lectin-horseradish peroxidase conjugates

Summary Fragments of mesonephros were taken from chick embryos and studied from the 4th to the 21st day of incubation. A battery of seven different horseradish peroxidase-labelled lectins was used to study the distribution of carbohydrate residues in glycoconjugates along the mesonephric nephron during the period of excretory activity and the period of involution. ConA and WGA reacted at every site of the nephron thus showing the ubiquitous presence of -D-mannose andN-acetyl-d-glucosamine. SBA was a good marker of the proximal tubule. Other lectins, such as PNA and LTA, reacted only for a short time at some sites during the considered period of incubation. The presence of sialic acid was detected in the podocytes, capillary wall and mesangial cells. From the 10th-11th day of incubation changes were noted in the proximal tubule as shown by PNA reactivity. This may be significant as regards the exact stage of incubation during which the involution of mesonephros begins.     

Quantitative immunogold localization of Na,K-ATPase along rat nephron

Summary Ultrastructural localization of Na, K-ATPase -subunit along rat nephron segments was investigated quantitatively by immunogold electron microscopy on LR-White ultrathin sections using affinity-purified antibody against -subunit of the enzyme. Ultrathin sections were incubated with the antibody at a saturation level and the number of gold particles bound per m of the plasma membrane (particle density) of the tubular epithelial cells from the proximal tubule to the collecting duct was determined. In all the tubular epithelial cells, gold particles were located exclusively on the basolateral surface, and no significant binding of gold particles to the apical surface was observed. Distribution of gold particles on the basolateral membranes was quite heterogeneous; lateral membranes and infolded basal membranes were highly labeled, whereas the basal membranes which are in direct contact with the basal lamina were scarcely labeled. The average particle density on the basal surface was highest in the distal straight tubule cells (11.4 units), very high in the distal convoluted tubule cells (9.8 units), intermediate in the proximal tubule cells (3.3 units), in the connecting tubule cells (4.3 units), and in the principal cells of the collecting duct (5.6–3.8 units), low in the thin limb of Henle's loop (1.0 unit), and at the control level in the intercalated cells in the connecting and collecting duct. The relative number of gold particles/mm nephron segment and the relative number of gold particles in the various nephron segments were calculated using quantitative morphological data. The estimated distribution profile of the former was in good agreement with the Na, K-ATPase activity profile in rat nephron, which was determined biochemically with a microenzymatic method.     

Purification and initial characterization of phycobiliproteins from the endophytic cyanobacterium of Azolla

The endophytic cyanobacterium, Anabaena azollae, isolated from laboratory cultures of Azolla caroliniana Willd., contains three spectroscopically distinct biliproteins. About 70% of the biliprotein is c-phycocyanin (_max 610 nm) and 13% is allophycocyanin (_max 647 nm, shoulder 620 nm). A third pigment corresponds to phycoerythrocyanin (_max 570 nm, shoulder 590 nm). In very dilute solutions of allophycocyanin, at constant pH and buffer strength, the 647 nm maximum disappears and a single _max occurs at 615–620 nm. The 647 nm absorption maximum reappears upon concentrating the dilute solution. Very dilute solutions of phycoerythrocyanin exhibit a broad peak between 570 and 590 nm. Absorption spectra of c-phycocyanin are not significantly altered upon dilution. Fluorescence emission maxima of phycoerythrocyanin, c-phycocyanin, and allophycocyanin occur at 630 nm, 643 nm and 660 nm respectively, using 540 nm excitation. Two subunits, of molecular weight 16,500 () and 20,600 (), are seen in c-phycocyanin upon dissociation with SDS. Dissociation of allophycocyanin and phycoerythrocyanin with SDS yields one sizeclass of subunits, with a molecular weight of about 17,500 for allophycocyanin and 18,000 for phycoerythrocyanin.Contribution No. 684 Offprint requests to: G. A. Peters     

Walking on a ‘peg leg’: extensor muscle activities and sensory feedback after distal leg denervation in cockroaches

Previous studies in insects demonstrated that leg coordination changes following complete ablation of distal limb segments. However, normal coordination was restored when small peg leg prostheses were attached to leg stumps to permit substrate contact. We have adapted this paradigm to preserve appropriate leg mass and inertia by severing all nerves and muscle tendons in the femur of the cockroach hind leg and converting the animals own limb into a peg leg. Recordings of muscle activities and leg movements before and after denervation showed that: (1) the peg leg is actively used in walking and regular bursts occur in motoneurons to leg extensor muscles; (2) driving of motoneuron activity is sufficient to produce fictive bursting in a muscle whose tendon (apodeme) is cut in the ablation; and (3) similar motoneuron activities are found in walking on an oiled glass surface, when the effects of body weight and mechanical coupling are minimized. When distal segments were completely severed in these preparations, leg use and muscle bursting were disrupted but could be restored if the stumps were pressed against the substrate. These results support the hypothesis that feedback from receptors in proximal leg segments indicating forces allows for active leg use in walking.     

Asparagine-linked sugar chains of erythrocyte membrane glycoproteins from Wiskott-Aldrich syndrome are not impaired

In both healthy controls and patients with Wiskott-Aldrich syndrome, the main oligosaccharide in asparagine-linked sugar chains of the membrane glycoproteins of erythrocytes was biantennary sugar chain with bisected G1cNAc (Gal₂-GlcNAc₂-Man₃-GlcNAc-GlcNAc-Fuc-GlcNAc_OT). Biantennary sugar chain with an-fucosyl residue linked at the proximal GlcNAc was seen but biantennary sugar chain without an-fucosyl residue at the proximal GlcNAc was not detected in each subject. There was no difference in quality and quantity of asparagine-linked sugar chains of erythrocyte membrane glycoproteins between healthy controls and the patients. These results suggest that asparagine-linked sugar chains in membrane glycoproteins of hematopoietic cells may not be impaired in Wiskott-Aldrich syndrome.     

Axillary artery injury combined with delayed brachial plexus palsy due to compressive hematoma in a young patient: a case report

Aim

To prove the possibility of axillary nerve conduction changes following shoulder subluxation due to hemiplegia, in order to investigate the usefulness of screening nerve conduction studies in patients with hemiplegia for finding peripheral neuropathy.

Methods

Forty-four shoulders of twenty-two patients with a first-time stroke having flaccid hemiplegia were tested, 43 ± 12 days after stroke onset. Wasting and weakness of the deltoid were present in the involved side. Motor nerve conduction latency and compound muscle action potential (CMAP) amplitude were measured along the axillary nerve, comparing the paralyzed to the sound shoulder. The stimulation was done at the Erb's point whilst the recording needle electrode was inserted into the deltoid muscle 4 cm directly beneath the lateral border of the acromion. Wilcoxon signed rank test was used to compare the motor conduction between the sound and the paralytic shoulder. Mann-Whitney test was used to compare between plegic and sound shoulder in each side.

Results

Mean motor nerve conduction latency time to the deltoid muscle was 8.49, SD 4.36 ms in the paralyzed shoulder and 5.17, SD 1.35 ms in the sound shoulder (p < 0.001). Mean compound muscle action potential (CMAP) amplitude was 2.83, SD 2.50 mV in the paralyzed shoulder and was 7.44, SD 5.47 mV in the sound shoulder (p < 0.001). Patients with right paralyzed shoulder compared to patients with right sound shoulder (p < 0.001, 1-sided for latency; p = 0.003, 1-sided for amplitude), and patients with left paralyzed shoulder compared to patients with left sound shoulder (p = 0.011, 1-sided for latency, p = 0.001, 1-sided for amplitude), support the same outcomes. The electro-physiological changes in the axillary nerve may appear during the first six weeks after stroke breakout.

Conclusion

Continuous traction of the axillary nerve, as in hypotonic shoulder, may affect the electro-physiological properties of the nerve. It most probably results from subluxation of the head of the humerus, causing demyelinization and even axonopathy. Slowing of the conduction velocities of the axillary nerve in the paralyzed shoulders may be related also to the lowering of the skin temperature and muscular atrophy in the same limb. The usefulness of routine screening nerve conduction studies in the shoulder of hemiplegic patients seems to be advocated.     

Biological Healing of Large Diaphyseal Deep-frozen Allograft Transplants

The biological healing of large, deep-frozen, diaphyseal allografts was studied in 24 cats. Deep-frozen allograft stored at –80°C was used to replace a large defect, at least two-thirds of the cat's tibial diaphysis. Intra-medullary rod fixation was used. The healing was studied at observation periods of 4, 6, 8, 12, 16, 24 and 36 weeks. Biological parameters studied included microangiography for revascularisation and fracture healing, bone resorption, new bone formation and callus encasement. Deep-frozen allografts united without difficulty by 12 weeks. However, minimal revascularisation of the allograft occurred even at 9 months. Likewise, minimal biological repair activity was seen even at 9 months as shown by the parameters of bone resorption, cortical new bone formation and callus encasement. Biologically, deep-frozen cortical allografts remained inert for a long period of time in the adult cat.     

Biliprotein assembly in the hemidiscoidal phycobilisomes of the thermophilic cyanobacterium Mastigocladus laminosus Cohn. Characterization of dissociation products with special reference to the peripheral phycoerythrocyanin-phycocyanin complexes

The dissociation products of isolated phycobilisomes of Mastigocladus laminosus were separated and analyzed by ultracentrifugation and, in part, by isoelectric focusing. With the exception of the allophycocyanin core, the sedimentation constants of peripheral phycocyanin- and phycoerythrocyanin-phycocyanin complexes lay in the range of 6 to 17S. The latter was represented by a 17S aggregate of two hexameric phycocyanins (dodecamer, dipartite unit). A complex with an absorption maximum at 610 nm (phycocyanin) and a shoulder at 580 nm (phycoerythrocyanin), a fluorescence emission maximum at 645 nm and a sedimentation constant of 11 S is described as a heterogeneously composed hexamer of ()₃-phycoerythrocyanin-()₃-phycocyanin. It was stable under extended dissociation in the cold and under isoelectric focusing. An aggregate of 14 S with an absorption maximum at 576 nm and a shoulder in the fluorescence emission spectrum at 625 nm (phycoerythrocyanin) in addition to the maximum at 645 nm (phycocyanin) is interpreted as a polar phycoerythrocyanin/ phycoerythrocyanin-phycocyanin complex. Combining these complexes with phycocyanin dodecamers creates peripheral rods of the phycobilisome. A proposal of the phycobiliprotein distribution within the phycobilisome of M. laminosus is presented.Abbreviations APC allophycocyanin - PC phycocyanin - PE phycoerythrin - PEC phycoerythrocyanin     

Asparagine-linked sugar chains of plasma membrane glycoproteins from healthy and common variable immunodeficiency B lymphoblastoid cell lines

Asparagine-linked sugar chains of plasma membrane glycoproteins, which are formed by glycosylation during B cell maturation, were examined with B lymphoblastoid cell lines (LCLs) transformed by Epstein-Barr virus derived from healthy controls and patients with common variable immunodeficiency (CVI). Both two patients with CVI showed hypogammaglobulinemia and impaired B cell functions. LCLs from healthy controls and the patients showed CD19+ and HLA/DR+ in the cell surface and secreted IgM. In both healthy controls and the patients, the main oligosaccharide in asparagine-linked sugar chains of the membrane glycoproteins of LCLs was biantennary sugar chain with bisected GlcNAc (Gal₂-GlcNAc₂-Man₃-GlcNAc-GlcNAc-Fuc-GlcNAc_OT). Biantennary sugar chain with an-fucosyl residue linked at the proximal GIcNAc was seen but biantennary sugar chain without an-fucosyl residue at the proximal GlcNAc was little detected in each LCL. There was no difference in quality and quantity of asparagine-linked sugar chains between healthy controls and the patients. These results suggest that glycosylation during B cell maturation may not be impaired in patients with CVI.