Estrogen depletion alters mineralization regulation mechanisms in an ovariectomized monkey animal model.

Ovariectomized animal models have been extensively used in osteoporosis research due to the resulting loss of bone mass. The purpose of the present study was to test the hypothesis that estrogen depletion alters mineralization regulation mechanisms in an ovariectomized monkey animal model. To achieve this we used Raman microspectroscopy to analyze humeri from monkeys that were either SHAM-operated or ovariectomized (N = 10 for each group). Measurements were made as a function of tissue age and cortical surface (periosteal, osteonal, endosteal) based on the presence of calcein fluorescent double labels. In the present work we focused on osteoid seams (defined as a surface with evident calcein labels, 1 µm distance away from the mineralizing front, and for which the Raman spectra showed the presence of organic matrix but not mineral), as well as the youngest mineralized tissue between the second fluorescent label and the mineralizing front, 1 µm inwards from the front with the phosphate mineral peak evident in the Raman spectra (TA1). The spectroscopically determined parameters of interest were the relative glycosaminoglycan (GAG) and pyridinoline (Pyd) contents in the osteoid, and the mineral content in TA1. At all three cortical surfaces, significant correlations were evident in the SHAM-operated animals between osteoid GAG (negative) and Pyd content, and mineral content, unlike the OVX animals. These results suggest that in addition to the well-established effects on turnover rates and bone mass, estrogen depletion alters the regulation of mineralization by GAGs and Pyd.

Epizootics of Streptococcus agalactiae infection in captive rays from Queensland, Australia.

The aim of this study was to describe two epizootics of high mortalities from infection with Streptococcus agalactiae, occurring in captive rays held in a marine display aquarium in south-east Queensland, Australia, in 2009 and 2010. Five different species of rays were affected, including mangrove whiprays (Himantura granulata), estuary rays (Dasyatis fluviorum), eastern shovelnose rays (Aptychotrema rostrata), white-spotted eagle rays (Aetobatus narinari) and blue-spotted mask rays (Neotrygon kuhlii). This report describes the history of both epizootics including collection, quarantine and husbandry of rays, the disease epizootics, clinico-pathological features of the disease, antimicrobial therapy, autogenous vaccine production, and laboratory studies including clinical and histopathology, bacteriology, PCR, molecular serotyping and sequencing of the bacterium S. agalactiae.

Genetic deletion of Sost or pharmacological inhibition of sclerostin prevent multiple myeloma-induced bone disease without affecting tumor growth.

Multiple myeloma (MM) causes lytic bone lesions due to increased bone resorption and concomitant marked suppression of bone formation. Sclerostin (Scl), an osteocyte-derived inhibitor of Wnt/ß-catenin signaling, is elevated in MM patient sera and increased in osteocytes in MM-bearing mice. We show here that genetic deletion of Sost, the gene encoding Scl, prevented MM-induced bone disease in an immune-deficient mouse model of early MM, and that administration of anti-Scl antibody (Scl-Ab) increased bone mass and decreases osteolysis in immune-competent mice with established MM. Sost/Scl inhibition increased osteoblast numbers, stimulated new bone formation and decreased osteoclast number in MM-colonized bone. Further, Sost/Scl inhibition did not affect tumor growth in vivo or anti-myeloma drug efficacy in vitro. These results identify the osteocyte as a major contributor to the deleterious effects of MM in bone and osteocyte-derived Scl as a promising target for the treatment of established MM-induced bone disease. Further, Scl did not interfere with efficacy of chemotherapy for MM, suggesting that combined treatment with anti-myeloma drugs and Scl-Ab should effectively control MM growth and bone disease, providing new avenues to effectively control MM and bone disease in patients with active MM.

Infection and pathology in Queensland grouper, Epinephelus lanceolatus, (Bloch), caused by exposure to Streptococcus agalactiae via different routes.

Since 2007, 96 wild Queensland groupers, Epinephelus lanceolatus, (Bloch), have been found dead in NE Australia. In some cases, Streptococcus agalactiae (Group B Streptococcus, GBS) was isolated. At present, a GBS isolate from a wild grouper case was employed in experimental challenge trials in hatchery-reared Queensland grouper by different routes of exposure. Injection resulted in rapid development of clinical signs including bilateral exophthalmia, hyperaemic skin or fins and abnormal swimming. Death occurred in, and GBS was re-isolated from, 98% fish injected and was detected by PCR in brain, head kidney and spleen from all fish, regardless of challenge dose. Challenge by immersion resulted in lower morbidity with a clear dose response. Whilst infection was established via oral challenge by admixture with feed, no mortality occurred. Histology showed pathology consistent with GBS infection in organs examined from all injected fish, from fish challenged with medium and high doses by immersion, and from high-dose oral challenge. These experimental challenges demonstrated that GBS isolated from wild Queensland grouper reproduced disease in experimentally challenged fish and resulted in pathology that was consistent with that seen in wild Queensland grouper infected with S. agalactiae.

Natural outbreak of Streptococcus agalactiae (GBS) infection in wild giant Queensland grouper, Epinephelus lanceolatus (Bloch), and other wild fish in northern Queensland, Australia.

Ninety-three giant Queensland grouper, Epinephelus lanceolatus (Bloch), were found dead in Queensland, Australia, from 2007 to 2011. Most dead fish occurred in northern Queensland, with a peak of mortalities in Cairns in June 2008. In 2009, sick wild fish including giant sea catfish, Arius thalassinus (Rüppell), and javelin grunter, Pomadasys kaakan (Cuvier), also occurred in Cairns. In 2009 and 2010, two disease epizootics involving wild stingrays occurred at Sea World marine aquarium. Necropsy, histopathology, bacteriology and PCR determined that the cause of deaths of 12 giant Queensland grouper, three wild fish, six estuary rays, Dasyatis fluviorum (Ogilby), one mangrove whipray, Himantura granulata (Macleay), and one eastern shovelnose ray, Aptychotrema rostrata (Shaw), was Streptococcus agalactiae septicaemia. Biochemical testing of 34 S. agalactiae isolates from giant Queensland grouper, wild fish and stingrays showed all had identical biochemical profiles. The 16S rRNA gene sequences of isolates confirmed all isolates were S. agalactiae; genotyping of selected S. agalactiae isolates showed the isolates from giant Queensland grouper were serotype Ib, whereas isolates from wild fish and stingrays closely resembled serotype II. This is the first report of S. agalactiae from wild giant Queensland grouper and other wild tropical fish and stingray species in Queensland, Australia.

Strontium ranelate does not stimulate bone formation in ovariectomized rats.

INTRODUCTION: Strontium ranelate (SrR) is suggested to function as a dual-acting agent in the treatment of postmenopausal osteoporosis with anti-resorptive and anabolic skeletal benefits. We evaluated the effects of SrR on the skeleton in ovariectomized (OVX) rats and evaluated the influence of dietary calcium. METHODS: Three-month old virgin female rats underwent ovariectomy (OVX, n = 50) or SHAM surgery (SHAM, n = 10). Four weeks post-surgery, rats were treated daily by oral gavage with distilled water (10 ml/kg/day) or SrR (25 or 150 mg/kg/day) for 90 days. Separate groups of animals for each dose of SrR were fed a low (0.1%) or normal (1.19%) calcium (Ca) diet. Static and dynamic histomorphometry, DXA, mu-CT, mechanical testing, and serum and skeletal concentrations of strontium were assessed. RESULTS: SrR at doses of 25 and 150 mg/kg/day did not increase bone formation on trabecular or periosteal bone surfaces, and failed to inhibit bone resorption of trabecular bone regardless of Ca intake. There were no improvements in bone mass, volume or strength with either dose of SrR given normal Ca. CONCLUSION: These findings demonstrate that SrR at dosages of 25 and 150 mg/kg/day did not stimulate an anabolic bone response, and failed to improve the bone biomechanical properties of OVX rats.

Germ-line transformation of the Mexican fruit fly.

Germ-line transformation of a major agricultural pest, the Mexican fruit fly (Anastrepha ludens Loew, Mexfly), was achieved using composite piggyBac transposable elements marked with green, yellow and red fluorescent proteins (CopGreen, PhiYFP and J-Red). We also investigated the possibility of generating transposon-free insertions, in order to address potential concerns relating to proposed field use of transgenic Mexfly. We describe a highly efficient method for transforming Mexfly, compare efficiency of piggyBac terminal sequences for transformation and also describe the derivation of a transposon-free insertion line. The development of an efficient transformation system for Mexfly holds great promise for improved applications of the sterile insect technique, a major component of the present control measures for this economically important pest species.

Detection of Chlamydiaceae DNA in veterinary specimens using a family-specific PCR.

AIMS: The aim of this work was to develop a rapid molecular test for the detection of the Chlamydiaceae family, irrespective of the species or animal host. METHODS AND RESULTS: The method described herein is a polymerase chain reaction targeting the 16S rRNA gene of the Chlamydiaceae family, and the results demonstrate that the test reacts with five reference Chlamydiaceae but none of the 19 other bacterial species or five uninfected animal tissues tested. The results also indicate the enhanced sensitivity of this test when compared with conventional culture or serology techniques. This is demonstrated through parallel testing of six real clinical veterinary cases and confirmatory DNA sequence analysis. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: This test can be used by veterinary diagnostic laboratories for rapid detection of Chlamydiaceae in veterinary specimens, with no restriction of chlamydial species or animal host. The test does not differentiate chlamydial species, and if required, speciation must be carried out retrospectively using alternate methods. However, for the purpose of prescribing therapy for chlamydiosis, this test would be an invaluable laboratory tool.

Involvement of MIIC-like late endosomes in B cell receptor-mediated antigen processing in murine B cells.

Currently, the involvement of classical vs novel endocytic compartments in the phenomenon of B cell receptor (BCR)-mediated Ag processing is a matter of considerable debate. In murine B cells, class II vesicles (CIIV) represent a novel endocytic compartment involved in BCR-mediated Ag processing and class II peptide loading. Alternatively, in human B cells, the MHC class II-enriched compartment (MIIC) represents a lysosome (L)-like endocytic compartment that appears to be involved in this process. Presently, the relationship between CIIV, MIIC, and classical endosomes and L remains to be determined. Using density gradient centrifugation, a subcellular compartment morphologically and immunologically similar to human MIIC has been identified, isolated, and characterized in murine B cells. These MIIC-like vesicles represent a population of class II-positive late endosomes (LE) and are distinct from CIIV. MIIC-like LE are uniquely marked by the thiol protease cathepsin B, and along with mature L, appear to be the major repository of DM molecules in these cells. Importantly, both MIIC-like LE and CIIV isolated from Ag-pulsed B cells contain BCR-internalized Ag as well as antigenic peptide-class II complexes.

Bilateral spontaneous rupture of flexor digitorum profundus tendons.

Spontaneous tendon rupture is an unusual condition usually associated with underlying disease processes such as rheumatoid arthritis, chronic renal failure or bony abnormalities of the hand. We report a case of spontaneous, non-concurrent bilateral rupture of flexor profundus tendons in an otherwise healthy individual. Treatment was successful and consisted of a two-stage reconstruction of the ruptured tendon.

Modification of native collagen reduces antigenicity but preserves cell compatibility.

Porcine intestinal collagen (ICL), derived from processed small intestine, is used as a part of a remodelable bilaminate biosynthetic vascular prosthesis. The process for the production of ICL involves mechanical cleaning of non-crosslinked porcine intestine (NC-ICL), disinfection with peracetic acid (PA-ICL), and crosslinking with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (PA/EDC-ICL). Two model systems were investigated to evaluate the effect of these agents on the humoral response to NC-ICL. First, the antibody titers of rabbits immunized with NC-ICL, PA-ICL, and PA/EDC-ICL were determined, and second, the humoral response of canines receiving collagenous vascular implants was examined. Collagenous and noncollagenous fractions were extracted from NC-ICL, PA-ICL, and PA/EDC-ICL and separated by SDS-PAGE. PA and EDC treatment decreased the number of extractable proteins as compared to NC-ICL. Immunoblot techniques demonstrated anti-NC-ICL antibodies recognized multiple immunoreactive proteins in NC-ICL, but not in PA-ICL or PA/EDC-ICL; and rabbits immunized with NC-ICL produced higher antibody titers to ICL proteins than rabbits immunized with either PA-ICL or PA/EDC-ICL. It was, therefore, apparent that NC-ICL was more antigenic than either PA-ICL or PA/EDC-ICL. The humoral immune response of canines to PA/EDC-ICL fabricated vascular grafts was determined. At 4 weeks, 8 weeks, and 12 weeks postimplant, serum antibodies to ICL proteins or type I collagen could not be detected. These data demonstrate a reduced humoral immune response to PA/EDC-ICL. (c) 1996 John Wiley & Sons, Inc.

The use of calcium alginate dressings in deep hand burns.

Deep burns of the hand are a common serious surgical problem with major occupational and economic implications. Control of haemorrhage during excision and grafting is difficult and postoperative haematoma may reduce graft take. Following excision, important deep structures such as extensor tendons or joints may be exposed. Such tissues do not take free skin grafts satisfactorily. We have found calcium alginate dressings can be of immense help in minimizing these technical problems. We describe our experience in one patient; three other patients have been treated with equal success.

Facial fractures in Gaelic football and hurling.

A one year, retrospective, epidemiological study of all facial fractures, sustained whilst playing the GAA sports of football and hurling, treated in the Cork Regional Hospital was undertaken. The results have been analysed and compared to a similar study performed in this unit in 1975. Of 332 patients treated for facial fractures, 110 (33%) were injured whilst playing sport and 47% of these occurred when playing Gaelic football or hurling (52 injuries in all). Eighty per cent of Gaelic football and hurling patients required operative treatment. All surgery was performed under general anaesthetic. The numbers of hurling fractures have more than halved since 1975-76. This coincides with an increase in the numbers hurling, an increase in the use of protective headgear and vastly improved coaching. This study demonstrates that improved safety can be achieved without diluting sporting enjoyment.

Value of wearing head protection gear while playing hurling.

One of the three national games of Ireland, hurling is a contact team sport (15 a side) played with a metre long ash stick and a small hard leather ball. Over a 12 month period, 413 players were treated for hurling-related injuries at Cork Regional Hospital. While hand and facial trauma predominate, the proportion between the two sites has changed substantially from previous reports with a reduction in the level of facial injuries. The wearing of a helmet, and optionally a supplementary faceguard, is seen to have contributed to this trend. Despite this welcome reduction of facial injuries, a certain resistance to the use of protective headgear is evident, particularly among older players.

Severe dog-bite injuries, introducing the concept of pack attack: a literature review and seven case reports.

Attacks on humans by dogs acting as a pack, though uncommon, result in severe, sometimes fatal, injuries. We report seven cases of attacks by packs of dogs (five on children and two on elderly women) including one fatal case. The dangers of dogs acting as a pack are highlighted, particularly when confronted with an unaccompanied child or elderly female. The pattern of injuries and principles of treatment are discussed.

Neonatal brain dopamine depletion and the cortical and behavioral consequences of enriched postweaning environment.

This study investigated the effects of neonatal intraventricular administration of 6-hydroxydopamine (6-OHDA, 15 micrograms total with and without desmethylimipramine pretreatment) on the cortical thickening and behavioral effects of 35 days of enriched postweaning housing (ENR) in the rat. The 6-OHDA treatment depleted cortical dopamine (DA) to about 40% of control. It did not affect the thickness of the cerebral cortex nor did it affect the capacity for the cortex to be thickened by ENR. In addition, it did not alter the superior performance on two spatial water maze tasks that was caused by ENR. Thus, the potential for neurobehavioral plasticity was not changed by neonatal DA depletion. ENR eliminated the spatial learning/memory deficits that were caused by neonatal DA depletion and that were manifested when the rat was raised in standard (impoverished) laboratory conditions. Hence, environmental factors can modulate the cognitive effects of neonatal DA depletion. ENR did not attenuate the hyperactivity of the neonatal DA-depleted rat. This may reflect the subcortical mediation of this behavioral abnormality.

The horn flap: a curved V-Y advancement flap with lateral pedicle.

The Horn Flap, a curved V-Y advancement flap with a radially based subcutaneous pedicle, has proven to be a versatile, reliable method of reconstruction following excision of small facial lesions, particularly in the medial canthal area. Entry of the pedicle on the radial, or concave, side makes significant advancement possible.

Nail re-growth in fingertip injuries.

One hundred and fifty-five fingertip injuries were treated over a one-year period. Seventy-six patients with 90 fingertip injuries attended for follow-up. Sixty-four of these injuries were suitable for study. In ten cases, the nail was either trimmed and left in-situ, or cleaned and replaced (Group A). In the remaining 54 cases, the nail was either missing, or had been removed at operation (Group B). The two groups were compared in terms of nail re-growth and nail appearance, but no statistically significant difference was found. There was a direct correlation between final outcome and the degree of crush injury. We conclude that it is the severity of the original injury and the accuracy of the primary repair of the nailbed which are the main determinants of the outcome of a nailbed injury. Non-replacement of the nail did not appear to adversely affect nail re-growth and nail appearance.