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A microfluidic chip for generation of gradients of dissolved oxygen was designed, fabricated and tested. The novel way of active oxygen depletion through a gas permeable membrane was applied. Numerical simulations for generation of O(2) gradients were correlated with measured oxygen concentrations. The developed microsystem was used to study growth patterns of the bacterium Pseudomonas aeruginosa in medium with different oxygen concentrations. The results showed that attachment of Pseudomonas aeruginosa to the substrate changed with oxygen concentration. This demonstrates that the device can be used for studies requiring controlled oxygen levels and for future studies of microaerobic and anaerobic conditions.

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Although treponemes are consistently identified in tissue from bovine digital dermatitis (DD) lesions, the definitive etiology of this debilitating polymicrobial disease is still unresolved. To study the microbiomes of 27 DD-infected and 10 healthy interdigital skin samples, we used a combination of different molecular methods. Deep sequencing of the 16S rRNA gene variable regions V1-V2 showed that Treponema, Mycoplasma, Fusobacterium and Porphyromonas were the genera best differentiating the DD samples from the controls. Additional deep sequencing analysis of the most abundant genus, Treponema, targeting another variable region of the 16S rRNA gene, V3-V4, identified 15 different phylotypes, among which Treponema phagedenis-like and Treponema refringens-like species were the most abundant. Although the presence of Treponema spp., Fusobacterium necrophorum and Porphyromonas levii was confirmed by fluorescence in situ hybridization (FISH), the results for Mycoplasma spp. were inconclusive. Extensive treponemal epidermal infiltration, constituting more than 90% of the total bacterial population, was observed in 24 of the 27 DD samples. F. necrophorum and P. levii were superficially located in the epidermal lesions and were present in only a subset of samples. RT-qPCR analysis showed that treponemes were also actively expressing a panel of virulence factors at the site of infection. Our results further support the hypothesis that species belonging to the genus Treponema are major pathogens of DD and also provide sufficient clues to motivate additional research into the role of M. fermentans, F. necrophorum and P. levii in the etiology of DD.

Hypercalcemia is defined as a serum calcium concentration that is greater than two standard deviations above the normal mean, which in children may vary with age and sex, reflecting changes in the normal physiology at each developmental stage. Hypercalcemic disorders in children may present with hypotonia, poor feeding, vomiting, constipation, abdominal pain, lethargy, polyuria, dehydration, failure to thrive, and seizures. In severe cases renal failure, pancreatitis and reduced consciousness may also occur and older children and adolescents may present with psychiatric symptoms. The causes of hypercalcemia in children can be classified as parathyroid hormone (PTH)‐dependent or PTH‐independent, and may be congenital or acquired. PTH‐independent hypercalcemia, ie, hypercalcemia associated with a suppressed PTH, is commoner in children than PTH‐dependent hypercalcemia. Acquired causes of PTH‐independent hypercalcemia in children include hypervitaminosis; granulomatous disorders, and endocrinopathies. Congenital syndromes associated with PTH‐independent hypercalcemia include idiopathic infantile hypercalcemia (IIH), William's syndrome, and inborn errors of metabolism. PTH‐dependent hypercalcemia is usually caused by parathyroid tumors, which may give rise to primary hyperparathyroidism (PHPT) or tertiary hyperparathyroidism, which usually arises in association with chronic renal failure and in the treatment of hypophosphatemic rickets. Acquired causes of PTH‐dependent hypercalcemia in neonates include maternal hypocalcemia and extracorporeal membrane oxygenation. PHPT usually occurs as an isolated nonsyndromic and nonhereditary endocrinopathy, but may also occur as a hereditary hypercalcemic disorder such as familial hypocalciuric hypercalcemia, neonatal severe primary hyperparathyroidism, and familial isolated primary hyperparathyroidism, and less commonly, as part of inherited complex syndromic disorders such as multiple endocrine neoplasia (MEN). Advances in identifying the genetic causes have resulted in increased understanding of the underlying biological pathways and improvements in diagnosis. The management of symptomatic hypercalcemia includes interventions such as fluids, antiresorptive medications, and parathyroid surgery. This article presents a clinical, biochemical, and genetic approach to investigating the causes of pediatric hypercalcemia. © 2017 The Authors. Journal of Bone and Mineral Research Published by Wiley Periodicals Inc.

The bacteria associated with the infectious claw disease bovine digital dermatitis (DD) are spirochetes of the genus Treponema; however, their environmental reservoir remains unknown. To our knowledge, the current study is the first report of the discovery and phylogenetic characterization of rRNA gene sequences from DD-associated treponemes in the dairy herd environment. Although the spread of DD appears to be facilitated by wet floors covered with slurry, no DD-associated treponemes have been isolated from this environment previously. Consequently, there is a lack of knowledge about the spread of this disease among cows within a herd as well as between herds. To address the issue of DD infection reservoirs, we searched for evidence of DDassociated treponemes in fresh feces, in slurry, and in hoof lesions by deep sequencing of the V3 and V4 hypervariable regions of the 16S rRNA gene coupled with identification at the operational-taxonomic-unit level. Using treponeme-specific primers in this high-throughput approach, we identified small amounts of DNA (on average 0.6% of the total amount of sequence reads) from DD-associated treponemes in 43 of 64 samples from slurry and cow feces collected from six geographically dispersed dairy herds. Species belonging to the Treponema denticola/Treponema pedis-like and Treponema phagedenis-like phylogenetic clusters were among the most prevalent treponemes in both the dairy herd environment and the DD lesions. By the high-throughput approach presented here, we have demonstrated that cow feces and environmental slurry are possible reservoirs of DD-associated treponemes. This method should enable further clarification of the etiopathogenesis of DD. Bovine digital dermatitis (DD) was first described in 1974 and has since become a growing problem worldwide (1). The disease is characterized by focal proliferative to ulcerative dermatitis that is typically located on the plantar aspect of the hoof (1). The consequences of this disease are decreased animal welfare and serious economic losses for the farmers due to factors such as reduced milk yield and premature culling (2-4). DD is considered a multifactorial disease, and moist, unhygienic conditions, such as the slurry to which the animals are frequently exposed, are considered predisposing factors (5, 6). The bacteria most often associated with DD are spirochetes of the genus Treponema, which predominate in the deepest regions of DD lesions (7,8). To date, at least 20 different phylotypes have been identified from DD biopsy specimens; among these, Treponema phagedenis-like, Treponema medium/Treponema vincentii-like, Treponema denticolalike, and Treponema pedis phylotypes are highly associated with progression of the disease (9-12). DD also appears to be highly infectious based on the rapid intra-and interherd spread of the disease (6, 13, 14). The spread of DD in dairy cattle is facilitated by wet walking surfaces covered with feces (15), although the causative agents have not yet been found in slurry (16). Because the Treponema ...

Systematic studies of autosomal dominant osteopetrosis (ADO) were followed by the identification of underlying mutations giving unique possibilities to perform translational studies. What was previously designated ADO1 turned out to be a high bone mass phenotype caused by a missense mutation in the first propeller of LRP5, a region of importance for binding inhibitory proteins. Thereby, ADO1 cannot be regarded as a classical form of osteopetrosis but must now be considered a disease of LRP5 activation. ADO (Albers-Schönberg disease, or previously ADO2) is characterized by increased number of osteoclasts and a defect in the chloride transport system (ClC-7) of importance for acidification of the resorption lacuna (a form of Chloride Channel 7 Deficiency Osteopetrosis). Ex vivo studies of osteoclasts from ADO have shown that cells do form normally but have reduced resorption capacity and an expanded life span. Bone formation seems normal despite decreased osteoclast function. Uncoupling of formation from resorption makes ADO of interest for new strategies for treatment of osteoporosis. Recent studies have integrated bone metabolism in whole-body energy homeostasis. Patients with ADO may have decreased insulin levels indicating importance beyond bone metabolism. There seems to be a paradigm shift in the treatment of osteoporosis. Targeting ClC-7 might introduce a new principle of dual action. Drugs affecting ClC-7 could be antiresorptive, still allowing ongoing bone formation. Inversely, drugs affecting the inhibitory site of LRP5 might stimulate bone formation and inhibit resorption. Thereby, these studies have highlighted several intriguing treatment possibilities, employing novel modes of action, which could provide benefits to the treatment of osteoporosis.

This section opens with a nationwide Danish study on the risk of fractures in prostate cancer, as well as assessing the impact of exposure to androgen deprivation. The authors found that there was a marked increase in the risk of fractures, especially of the hip. Authors from the USA address the outcome of cytoreductive nephrectomy for metastatic RCC, finding that it can be predicted by the fraction of tumour volume removed. They give an informative review of the topic and describe their experience in this type of surgery. Treatment options for hormonerefractory prostate cancer are rather limited and authors from the USA present the results of a phase II trial into the use of gefitinib as a single therapy in patients with non‐metastatic refractory disease. They found an absence of PSA response but confirmed the well‐established favourable tolerability profile of this agent. OBJECTIVE To assess the risk of fracture attributable to prostate cancer, and the impact of exposure to prescribed gonadotrophin‐releasing hormone agonists and antiandrogens on this risk in a nationwide, population‐based case‐control study. PATIENTS AND METHODS Data from the Danish National Hospital Discharge Register, the National Bureau of Statistics, and the National Prescriptions Database were merged. The analysis covered 15 716 men aged >50 years presenting with a fracture at any hospital in Denmark in 2000, and 47 149 age‐matched control men. A previous diagnosis of prostate cancer had been recorded in 1.3% of controls and 2.5% of those with a fracture. RESULTS Prostate cancer was associated with an increased odds ratio (95% confidence interval) for all fractures of 1.8 (1.6–2.1), for hip fractures of 3.7 (3.1–4.4), but no increased risk of vertebral fractures. The increased fracture risk became apparent early after diagnosis and remained pronounced even in long‐term survivors. Androgen deprivation therapy (ADT) with an odds ratio of 1.7  (1.2–2.5; P < 0.01) and orchidectomy, at 1.7 (1.2–2.4; P < 0.01) added to the overall fracture risk. In all, 3.1% of hip fractures in Danish men aged >50 years are attributable to prostate cancer. CONCLUSION Prostate cancer, orchidectomy and the use of ADT are associated with a markedly greater risk of fractures, especially of the hip. The risk of hip fracture is not confined to the very old, neither is the risk made negligible by the excess mortality in patients with advanced prostate cancer.