Current thinking: return to play and transient quadriplegia.
Athletes that participate in contact and collision sports assume risk of serious injury each time they take the field. For those athletes that have sustained an episode of transient quadriplegia, the decision of whether to return to competition can be a difficult one. Some athletes, realizing how close they may have come to permanent injury, may decide that further participation is not in their best interest. Others may be somewhat undecided, and some may want to return at all costs. As the treating physician, the goal is to identify those athletes who after a single episode of transient quadriplegia are at increased risk for further injury and consequently should discontinue participation in contact sports. Factors that may contribute to that determination include mechanism of injury, prior history of neurologic symptoms or injury, and anatomic features that may predispose to further injury such as disc herniation, fracture, or cervical stenosis.


Porous silicon-graphene oxide core-shell nanoparticles for targeted delivery of siRNA to the injured brain.
We report the synthesis, characterization, and assessment of a nanoparticle-based RNAi delivery platform that protects siRNA payloads against nuclease-induced degradation and efficiently delivers them to target cells. The nanocarrier is based on biodegradable mesoporous silicon nanoparticles (pSiNPs), where the voids of the nanoparticles are loaded with siRNA and the nanoparticles are encapsulated with graphene oxide nanosheets (GO-pSiNPs). The graphene oxide encapsulant delays release of the oligonucleotide payloads in vitro by a factor of 3. When conjugated to a targeting peptide derived from the rabies virus glycoprotein (RVG), the nanoparticles show 2-fold greater cellular uptake and gene silencing. Intravenous administration of the nanoparticles into brain-injured mice results in substantial accumulation specifically at the site of injury.


Diagnostic utility of neutrophil CD64 as a marker for early-onset sepsis in preterm neonates.
Neutrophil CD64 has been proposed as an early marker of sepsis. This study aims to evaluate the diagnostic utility of neutrophil CD64 for identification of early-onset sepsis in preterm neonates. The prospective study was conducted in a neonatal intensive care unit between November 2010 and June 2011. Preterm neonates in whom infection was suspected when they were <12 hours of age were enrolled. Complete blood count with differential, blood culture, neutrophil CD11b and CD64 measurement were performed. Receiver operating characteristic curve analysis was performed to evaluate the performance of neutrophil CD64 as biomarker of sepsis. A total of 158 preterm neonates was enrolled, 88 of whom were suspected infection. The suspected sepsis group was of lesser gestational age (P<0.001) and lower birth weight (P<0.001), compared with controls. The hematologic profiles of the suspected sepsis group were characterized by higher white blood cell count, neutrophil counts and C-reactive protein. The suspected sepsis neonates had significantly higher neutrophil CD64 expression compared with controls. Neutrophil CD64 had an area value under the curve of 0.869 with an optimal cutoff values of 1010 phycoerythrin molecules bound/cell and it had a high sensitivity (81.82%) and negative predictive value (77.4%). The level of neutrophil CD64 was independent of antibiotic therapy within 24 hours after the onset of sepsis in preterm neonates. Neutrophil CD64 is a highly sensitive marker for suspected early-onset sepsis in preterm neonates. Our study suggests that neutrophil CD64 may be incorporated as a valuable marker to diagnose infection.


Leptin and hyperleptinemia - from friend to foe for cardiovascular function.
The obese gene product, leptin, plays a central role in food intake and energy metabolism. The physiological roles of leptin in human bodily function have been broadened over the past decade since leptin was first discovered in 1994. Evidence has suggested that leptin plays a specific role in the intricate cascade of cardiovascular events, in addition to its well-established metabolic effects. Leptin, a hormone linking adiposity and central nervous circuits to reduce appetite and enhance energy expenditure, has been shown to increase overall sympathetic nerve activity, facilitate glucose utilization and improve insulin sensitivity. In addition, leptin is capable of regulating cardiac and vascular contractility through a local nitric oxide-dependent mechanism. However, elevated plasma leptin levels or hyperleptinemia, have been demonstrated to correlate with hyperphagia, insulin resistance and other markers of the metabolic syndrome including obesity, hyperlipidemia and hypertension, independent of total adiposity. Elevated plasma leptin levels may be an independent risk factor for the development of cardiovascular disease. Although mechanisms leading to hyperleptinemia have not been well described, factors such as increased food intake and insulin resistance have been shown to rapidly enhance plasma leptin levels and subsequently tissue leptin resistance. These findings have prompted the speculation that leptin in the physiological range may serve as a physiological regulator of cardiovascular function whereas elevated plasma leptin levels may act as a pathophysiological trigger and/or marker for cardiovascular diseases due to tissue leptin resistance.


Analysis of LOXL1 polymorphisms in a United States population with pseudoexfoliation glaucoma.
To identify if recently described LOXL1 (lysyl oxidase-like 1) polymorphisms are associated with pseudoexfoliation glaucoma (XFG) in a United States (U.S.) Caucasian patient population. Individuals with XFG were identified using standard clinical examination techniques. TaqMan allelic discrimination assays were used to genotype 13 single nucleotide polymorphisms (SNPs) that tag LOXL1 in Caucasian individuals. The coding region of exon 1 that includes the previously associated SNP, rs1048661, was sequenced. Allele and genotype frequencies were compared between cases and unrelated controls. Fifty affected individuals and 235 control individuals were recruited into this study. We replicated the previously reported association of three SNPs (rs1048661, rs2165241, and rs3825942) in our independent XFG population (single SNP p-values were 0.001-0.02). The risk alleles at these three and several other intragenic SNPs are part of an extended XFG-associated LOXL1 haplotype with a frequency of 32.0% in XFG patients and 21.6% in controls. We have performed an analysis of LOXL1 and XFG in a United States patient population and have confirmed the strong association previously reported for Icelandic and Swedish samples. However, due to the high frequency of risk alleles in non-XFG individuals, this association should not form the basis of a diagnostic test for XFG. It is likely that additional genetic or environmental factors modulate the penetrance of LOXL1 susceptibility alleles.


Human bocaviruses are highly diverse, dispersed, recombination prone, and prevalent in enteric infections.
A new species of parvovirus, tentatively named human bocavirus 4 (HBoV4), was genetically characterized. Among 641 feces samples obtained from children and adults, the most commonly detected bocavirus species were, in descending order, HBoV2, HBoV3, HBoV4, and HBoV1, with an HBoV2 prevalence of 21% and 26% in Nigerian and Tunisian children, respectively. HBoV3 or HBoV4 species were found in 12 of 192 patients with non-polio acute flaccid paralysis in Tunisia and Nigeria and 0 of 96 healthy Tunisian contacts (P = .01). Evidence of extensive recombination at the NP1 and VP1 gene boundary between and within bocavirus species was found. The high degree of genetic diversity seen among the human bocaviruses found in feces specimens, relative to the highly homogeneous HBoV1, suggest that this worldwide-distributed respiratory pathogen may have recently evolved from an enteric bocavirus after acquiring an expanded tropism favoring the respiratory tract. Elucidating the possible role of the newly identified enteric bocaviruses in human diseases, including acute flaccid paralysis and diarrhea, will require further epidemiological studies.


GM-CSF action in the CNS decreases food intake and body weight.
Many proinflammatory cytokines, such as leptin, play key roles in dynamic regulation of energy expenditure and food intake. The present work tested a role for the proinflammatory cytokine GM-CSF. Central but not peripheral administration of GM-CSF to adult rats significantly decreased food intake and body weight for at least 48 hours. Similar results were observed following central administration of GM-CSF in mice. GM-CSF receptor immunoreactivity was found on neurons within the paraventricular and arcuate nuclei of the hypothalamus. GM-CSF-deficient (GM-/-) mice weighed more and had significantly higher total body fat than wild-type (GM+/+) mice. Energy expenditure in GM-/- mice was decreased compared with that in GM+/+ mice. Taken together, these findings demonstrate that GM-CSF signaling in the CNS can regulate energy homeostasis.


Bevacizumab and cyclosphosphamide, doxorubicin, vincristine and prednisone in combination for patients with peripheral T-cell or natural killer cell neoplasms: an Eastern Cooperative Oncology Group study (E2404).
Peripheral T-cell lymphoma (PTCL) and natural killer (NK) cell lymphoma have poor survival with conventional cytotoxic chemotherapy. Because angiogenesis plays an important role in the biology of PTCL, a fully humanized anti-vascular endothelial growth factor (VEGF) antibody, bevacizumab (A), was studied in combination with standard cyclosphosphamide, doxorubicin, vincristine and prednisone (CHOP) chemotherapy (ACHOP) to evaluate its potential to improve outcome in these patients. Patients were treated with 6-8 cycles of ACHOP followed by eight doses of maintenance A (15 mg/kg every 21 days). Forty-six patients were enrolled on this phase 2 study from July 2006 through March 2009. Forty-four patients were evaluable for toxicity and 39 were evaluable for response, progression and survival. A total of 324 cycles (range: 2-16, median 7) were administered to 39 evaluable patients and only nine completed all planned treatment. The overall response rate was 90% with 19 (49%) complete response/complete response unconfirmed (CR/CRu) and 16 (41%) a partial response (PR). The 1-year progression-free survival (PFS) rate was 44% at a median follow-up of 3 years. The median PFS and overall survival (OS) rates were 7.7 and 22 months, respectively. Twenty-three patients died (21 from lymphoma, two while in remission). Grade 3 or 4 toxicities included febrile neutropenia (n = 8), anemia (n = 3), thrombocytopenia (n = 5), congestive heart failure (n = 4), venous thrombosis (n = 3), gastrointestinal hemorrhage/perforation (n = 2), infection (n = 8) and fatigue (n = 6). Despite a high overall response rate, the ACHOP regimen failed to result in durable remissions and was associated with significant toxicities. Studies of novel therapeutics are needed for this patient population, whose clinical outcome remains poor.


Diffusion patterns of new anti-diabetic drugs into hospitals in Taiwan: the case of thiazolidinediones for diabetes.
Diffusion of new drugs in the health care market affects patients' access to new treatment options and health care expenditures. We examined how a new drug class for diabetes mellitus, thiazolidinediones (TZDs), diffused in the health care market in Taiwan. Assuming that monthly hospital prescriptions of TZDs could serve as a micro-market to perform drug penetration studies, we retrieved monthly TZD prescription data for 580 hospitals in Taiwan from Taiwan's National Health Insurance Research Database for the period between March 1, 2001 and December 31, 2005. Three diffusion parameters, time to adoption, speed of penetration (monthly growth on prescriptions), and peak penetration (maximum monthly prescription) were evaluated. Cox proportional hazards model and quantile regressions were estimated for analyses on the diffusion parameters. Prior hospital-level pharmaceutical prescription concentration significantly deterred the adoption of the new drug class (HR: 0.02, 95%CI = 0.01 to 0.04). Adoption of TZDs was slower in district hospitals (HR = 0.43, 95%CI = 0.24 to 0.75) than medical centers and faster in non-profit hospitals than public hospitals (HR = 1.79, 95%CI = 1.23 to 2.61). Quantile regression showed that penetration speed was associated with a hospital's prior anti-diabetic prescriptions (25%Q: 18.29; 50%Q: 25.57; 75%Q: 30.97). Higher peaks were found in hospitals that had adopted TZD early (25%Q: -40.33; 50%Q: -38.65; 75%Q: -32.29) and in hospitals in which the drugs penetrated more quickly (25%Q: 16.53; 50%Q: 24.91; 75%Q: 31.50). Medical centers began to prescribe TZDs earlier, and they prescribed more TZDs at a faster pace. The TZD diffusion patterns varied among hospitals depending accreditation level, ownership type, and prescription volume of Anti-diabetic drugs.


Effect of Acetabular Component Positioning on Functional Outcomes in Primary Total Hip Arthroplasty.
Acetabular component positioning is crucial to a successful total hip arthroplasty (THA). This study evaluated the effect of absolute acetabular component position as well as acetabular position relative to bony anatomy on patient-reported functional outcomes after primary THA. Primary, press fit, hemispherical metal-on-polyethylene THA performed between 2003 and 2011 were analyzed. Western Ontario and McMaster Osteoarthritis Index (WOMAC), Harris Hip Score (HHS), Short Form-12 scores, and radiographs between 2 and 3 years after the index procedure were assessed. Of the 1241 primary THA included, the mean abduction and anteversion angles were 44.4 ± 6.94 and 21.7 ± 11.9 degrees, respectively. The mean anterior and lateral overhang were 1.9 ± 3.6 and 2.5 ± 3.4 mm, respectively. There was no correlation between functional outcomes and acetabular inclination. A weak positive correlation between anteversion and HHS (P < .001) and WOMAC (P = .02) scores was found. For relative position, anterior overhang of the acetabular component beyond the bone resulted in inferior Short Form-12 physical function (P = .001), HHS (P = .004), and WOMAC (P < .001) scores compared to those with bony coverage. Mean HHS pain score was 41.20 ± 5.69 in patients with lateral overhang and 41.97 ± 5.04 in those who had bony coverage of the lateral edge of the acetabular component (P = .02). The tribology and biomechanics of acetabular component position have been extensively studied without examination of how it affects patient function. Although statistical significance was seen, clinical outcome scores were not sensitive enough to show a clinically significant effect of the absolute or relative position of the acetabular component.


Hydrocephalus in children with ruptured cerebral arteriovenous malformation.
Hydrocephalus is a strong determinant of poor neurological outcome after intracerebral hemorrhage (ICH). In children, ruptured brain arteriovenous malformations (bAVMs) are the dominant cause of ICH. In a large prospective cohort of pediatric patients with ruptured bAVMs, the authors analyzed the rates and predictive factors of hydrocephalus requiring acute external ventricular drainage (EVD) or ventriculoperitoneal shunt (VPS). The authors performed a single-center retrospective analysis of the data from a prospectively maintained database of children admitted for a ruptured bAVM since 2002. Admission clinical and imaging predictors of EVD and VPS placement were analyzed using univariate and multivariate statistical models. Among 114 patients (mean age 9.8 years) with 125 distinct ICHs due to ruptured bAVM, EVD and VPS were placed for 55/125 (44%) hemorrhagic events and 5/114 patients (4.4%), respectively. A multivariate nominal logistic regression model identified low initial Glasgow Coma Scale (iGCS) score, hydrocephalus on initial CT scan, the presence of intraventicular hemorrhage (IVH), and higher modified Graeb Scale (mGS) score as strongly associated with subsequent need for EVD (all p < 0.001). All children who needed a VPS had initial hydrocephalus requiring EVD and tended to have higher mGS scores. In a large cohort of pediatric patients with ruptured bAVM, almost half of the patients required EVD and 4.4% required permanent VPS. Use of a low iGCS score and a semiquantitative mGS score as indicators of the IVH burden may be helpful for decision making in the emergency setting and thus improve treatment.


Oxidative stress induces apoptosis via calpain- and caspase-3-mediated cleavage of ATM in pancreatic acinar cells.
Oxidative stress-induced DNA cleavage and apoptosis in pancreatic acinar cells has been implicated in the pathogenesis of acute pancreatitis. Thus, an efficient DNA repair process is key to prevention of apoptotic pancreatic acinar cell death. Ataxia telangiectasia mutated (ATM), a sensor of DNA breaks, functions by recruiting DNA repair proteins to initiate the DNA repair process. In the present study, we investigated whether H2O2 produced by the action of glucose oxidase on α-D-glucose (G/GO) induces apoptosis in pancreatic acinar AR42J cells through an alteration of the level of ATM. As a result, G/GO induced apoptosis by promoting a loss of cell viability, increase in Bax, decrease in Bcl-2, cleavage of poly (ADP-ribose) polymerase (PARP) and fragmentation of DNA. In addition, ATM cleavage along with elevated levels of calpain and caspase-3 activity was induced by G/GO. By using ATM siRNA, we demonstrated that reduction in ATM levels enhanced G/GO-induced apoptosis. Moreover, inhibition of calpain activity by calpeptin or calpastatin, or by inhibition of caspase-3 with z-DEVD, suppressed G/GO-induced apoptosis and ATM cleavage. Collectively, these findings suggest that proteolysis of ATM is the underlying mechanism of apoptosis of pancreatic acinar cells caused by exposure to oxidative stress.


The discovery of the virulence gene ToxA in the wheat and barley pathogen Bipolaris sorokiniana.
Bipolaris sorokiniana is the causal agent of multiple diseases on wheat and barley and is the primary constraint to cereal production throughout South Asia. Despite its significance, the molecular basis of disease is poorly understood. To address this, the genomes of three Australian isolates of B. sorokiniana were sequenced and screened for known pathogenicity genes. Sequence analysis revealed that the isolate BRIP10943 harboured the ToxA gene, which has been associated previously with disease in the wheat pathogens Parastagonospora nodorum and Pyrenophora tritici-repentis. Analysis of the regions flanking ToxA within B. sorokiniana revealed that it was embedded within a 12-kb genomic element nearly identical to the corresponding regions in P. nodorum and P. tritici-repentis. A screen of 35 Australian B. sorokiniana isolates confirmed that ToxA was present in 12 isolates. Sequencing of the ToxA genes within these isolates revealed two haplotypes, which differed by a single non-synonymous nucleotide substitution. Pathogenicity assays showed that a B. sorokiniana isolate harbouring ToxA was more virulent on wheat lines that contained the sensitivity gene when compared with a non-ToxA isolate. This work demonstrates that proteins that confer host-specific virulence can be horizontally acquired across multiple species. This acquisition can dramatically increase the virulence of pathogenic strains on susceptible cultivars, which, in an agricultural setting, can have devastating economic and social impacts.


Optimization of planar self-collimating photonic crystals.
Self-collimation in photonic crystals has received a lot of attention in the literature, partly due to recent interest in silicon photonics, yet no performance metrics have been proposed. This paper proposes a figure of merit (FOM) for self-collimation and outlines a methodical approach for calculating it. Performance metrics include bandwidth, angular acceptance, strength, and an overall FOM. Two key contributions of this work include the performance metrics and identifying that the optimum frequency for self-collimation is not at the inflection point. The FOM is used to optimize a planar photonic crystal composed of a square array of cylinders. Conclusions are drawn about how the refractive indices and fill fraction of the lattice impact each of the performance metrics. The optimization is demonstrated by simulating two spatially variant self-collimating photonic crystals, where one has a high FOM and the other has a low FOM. This work gives optical designers tremendous insight into how to design and optimize robust self-collimating photonic crystals, which promises many applications in silicon photonics and integrated optics.


Evaluating an insurer-based health coaching program: Impact of program engagement on healthcare utilization and weight loss.
Insurers and employers are increasingly offering lifestyle and weight-loss coaching programs; however, few evaluations have examined their effectiveness. Our objectives were to determine whether level of program engagement was associated with differences in healthcare utilization and weight pre/post coaching. We conducted a retrospective evaluation of enrollees in an insurer-based telephonic health coaching program in Maryland (2013-2014). Our independent variables were program engagement benchmarks (≥3 and ≥6 sessions). Our dependent variables included change in outpatient and emergency department (ED) visits (more visits post program, fewer visits post, or no change pre-post) and associated costs (difference pre-post) using claims data. We calculated mean percent weight change from baseline. We used multivariate-adjusted linear and multinomial logistic regression, as appropriate, to examine the association between outcomes and engagement benchmarks. We included 225 enrollees with mean age 50.7 years, 81.3% women, and mean body mass index of 35.0 kg/m2. Most participants focused on weight management (75.6%) and improving general health (57.8%). Few individuals had outpatient or ED visits, and no significant changes in healthcare utilization were associated with program engagement. Among the weight management subgroup (n = 170), mean weight change was -2.1% (SD 5.1). Participants achieved significantly greater weight loss if they met the 6-session engagement benchmark (β -3.5%, p < 0.01). Weight management is a popular focus for health coaching participants, and these programs can achieve modest weight loss. Programs should consider designing and testing strategies that promote engagement, given that weight-loss success was improved if participants completed at least 6 coaching sessions.


Posterior fossa craniotomy for lesions of the cerebellopontine angle. Technical note.
A safe technique is described for performing a lateral posterior fossa craniotomy to gain access to the cerebellopontine angle. The method makes use of currently available high-speed air drills. Thus, it is possible to replace the removed bone at the conclusion of the procedure and to re-establish normal tissue planes while providing rigid protection to the posterior fossa.


RNase H cleavage of the 5' end of the human immunodeficiency virus type 1 genome.
The synthesis of retroviral DNA is initiated near the 5' end of the RNA. DNA synthesis is transferred from the 5' end to the 3' end of viral RNA in an RNase H-dependent step. In the case of human immunodeficiency virus type 1 (HIV-1) (and certain other retroviruses that have complex secondary structures at the ends of the viral RNA), there is the possibility that DNA synthesis can lead to a self-priming event that would block viral replication. The extent of RNase H cleavage must be sufficient to allow the strand transfer reaction to occur, but not so extensive that self-priming occurs. We have used a series of model RNA substrates, with and without a 5' cap, to investigate the rules governing RNase H cleavage at the 5' end of the HIV-1 genome. These in vitro RNase H cleavage reactions produce an RNA fragment of the size needed to block self-priming but still allow strand transfer. The cleavages seen in vitro can be understood in light of the structure of HIV-1 reverse transcriptase in a complex with an RNA/DNA substrate.


Equity in use of home-based or facility-based skilled obstetric care in rural Bangladesh: an observational study.
Few studies have assessed whether the poorest people in developing countries benefit from giving birth at home rather than in a facility. We analysed whether socioeconomic status results in differences in the use of professional midwives at home and in a basic obstetric facility in a rural area of Bangladesh, where obstetric care was free of charge. We routinely obtained data from Matlab, Bangladesh between 1987 and 2001. We compared the benefits of home-based and facility-based obstetric care using a multinomial logistic and binomial log link regression, controlling for multiple confounders. Whether or not a midwife was used at home or in a facility differed significantly with wealth (adjusted odds ratio comparing the wealthiest and poorest quintiles 1.94 [95% CI 1.69-2.24] for home-based care, and 2.05 [1.72-2.43] for facility-based care). The gap between rich and poor widened after the introduction of facility-based care in 1996. The risk ratio (RR) between the wealthiest and poorest quintiles was 1.91 (adjusted RR 1.49 [95% CI 1.16-1.91] when most births with a midwife took place at home compared with 2.71 (1.66 [1.41-1.96]) at the peak of facility-based care. In this area of Bangladesh, a shift from home-based to facility-based basic obstetric care is feasible but might lead to increased inequities in access to health care. However, there is also evidence of substantial inequities in home births. Before developing countries reinforce home-based births with a skilled attendant, research is needed to compare the feasibility, cost, effectiveness, acceptability, and implications for health-care equity in both approaches.


Biobanking, consent, and commercialization in international genetics research: the Type 1 Diabetes Genetics Consortium.
and This article describes several ethical, legal, and social issues typical of international genetics biobanking, as encountered in the Type 1 Diabetes Genetics Consortium (T1DGC). By studying the examples set and lessons learned from other international biobanking studies and by devoting considerable time and resources to identifying, addressing, and continually monitoring ethical and regulatory concerns, T1DGC was able to minimize the problems reported by some earlier studies. Several important conclusions can be drawn based on the experience in this study: (1) Basic international standards for research ethics review and informed consent are broadly consistent across developed countries. (2) When consent forms are adapted locally and translated into different languages, discrepancies are inevitable and therefore require prompt central review and resolution before research is initiated. (3) Providing separate 'check-box' consent for different elements of a study creates confusion and may not be essential. (4) Creating immortalized cell lines to aid future research is broadly acceptable, both in the US and internationally. (5) Imposing some limits on the use of stored samples aids in obtaining ethics approvals worldwide. (6) Allowing potential commercial uses of donated samples is controversial in some Asian countries. (7) Obtaining government approvals can be labor-intensive and time-consuming, and can require legal and diplomatic skills.


Attachment and mental health symptoms among U.S. Afghanistan and Iraq veterans seeking health care services.
Attachment theory has become a primary framework for understanding adjustment to traumas. In a convenience sample of 157 U.S. service members from the Afghanistan and Iraq wars seeking health care services at a Veterans Administration (VA) hospital, this study examined (a) the impact of attachment characteristics on several key mental health symptoms in this new generation of veterans, (b) the relative frequencies of prominent attachment styles in the sample, and (c) how these higher order orientations related to study outcomes. First, with demographic and military background factors in the model, attachment-related anxiety and avoidance were each uniquely associated with posttraumatic stress symptoms (PTSS), psychiatric distress, and alcohol misuse, βs = .25 to .60. Second, latent class analysis highlighted the underrepresentation of avoidant veterans of a dismissive type in the sample. Third, of the different possible types of attachment insecurities, veterans with a fearful disorganized orientation were also shown to be particularly vulnerable to PTSS and other problems, even when compared to preoccupied-dependent persons. These findings yield further support for the protective benefit of attachment security, while also suggesting the importance of both attachment anxiety and avoidance in the context of postdeployment adjustment.


Future perspectives in bioreactor development.
The paper discusses conversion capacities of both anaerobic and aerobic wastewater treatment systems in relation to growth kinetics, hydrodynamics and biomass concentration. In the current modern anaerobic high-rate reactors the conversion potentials are optimally exploited. This is not yet true for aerobic systems since operation of aerobic systems under conditions of low biomass growth reduces the maximum applicable loading rates significantly. Both the concept of granulation and the introduction of fluidised bed systems have increased conversion capacities for both anaerobic and aerobic systems significantly. One of the latest development concerns the SBR with granular biomass. The grazing concept, in which ciliates convert aerobically grown dispersed cells, offers a possibility for significant improvement of aerobic systems. In the fields of psychrophilic and thermophilic anaerobic treatment, specific reactor development may contribute to further enhance volumetric conversion capacities. Due to reduced water usage, both COD and salt concentrations tend to increase for industrial effluents. As a consequence, there is a need for the development of anaerobic reactors retaining flocculant biomass. The membrane bioreactors offer a solution for certain niches in wastewater treatment. However the oxygen transfer economy is poor. There is a need for fundamental knowledge development to obtain a realistic image of this technology.


Assigning function to natural allelic variation via dynamic modeling of gene network induction.
More and more natural DNA variants are being linked to physiological traits. Yet, understanding what differences they make on molecular regulations remains challenging. Important properties of gene regulatory networks can be captured by computational models. If model parameters can be "personalized" according to the genotype, their variation may then reveal how DNA variants operate in the network. Here, we combined experiments and computations to visualize natural alleles of the yeast GAL3 gene in a space of model parameters describing the galactose response network. Alleles altering the activation of Gal3p by galactose were discriminated from those affecting its activity (production/degradation or efficiency of the activated protein). The approach allowed us to correctly predict that a non-synonymous SNP would change the binding affinity of Gal3p with the Gal80p transcriptional repressor. Our results illustrate how personalizing gene regulatory models can be used for the mechanistic interpretation of genetic variants.


To be on the safe site - Ungroomed spots on the bee's body and their importance for pollination.
Flower-visiting bees collect large quantities of pollen to feed their offspring. Pollen deposited in the bees' transport organs is lost for the flowers' pollination. It has been hypothesised that specific body areas, bees cannot groom, serve as 'safe sites' for pollen transfer between flowers. For the first time, we experimentally demonstrated the position, area and pollen amount of safe sites at the examples of Apis mellifera and Bombus terrestris by combining artificial contamination of the bees' body with pine or sunflower pollen and the subsequent bees' incomplete grooming. We found safe sites on the forehead, the dorsal thorax and waist, and on the dorsal and ventral abdomen of the bees. These areas were less groomed by the bees' legs. The largest amount of pollen was found on the waist, followed by the dorsal areas of thorax and abdomen. At the example of Salvia pratensis, S. officinalis and Borago officinalis, we experimentally demonstrated with fluorescent dye that the flowers' pollen-sacs and stigma contact identical safe sites. These results confirm that pollen deposition on the bees' safe sites improves pollen transfer to stigmas of conspecific flowers sti. Future research will demonstrate the importance of safe sites for plant pollination under field conditions.


Efficacy of subthreshold micropulse laser in the treatment of diabetic macular edema is influenced by pre-treatment central foveal thickness.
To determine if the severity of diabetic macular edema influences the effectiveness of subthreshold micropulse (STMP) laser treatment. A total of 63 eyes of 58 patients with diabetic macular edema were divided into two groups based on their initial central foveal thickness (CFT). Group 1 had CFT ≤400 μm, group 2 had CFT >400 μm. The change from baseline in CFT and visual acuity were compared at 3, 6 and 12 months follow-up. Patients were considered for retreatment with micropulse laser at 3 months if macular edema had not improved. Patients were considered for rescue anti-VEGF injections if there was clinically significant macular edema at 6 months follow-up. Number of laser retreatments, injections, and any adverse effects from STMP laser were recorded. Group 1 (n=33) experienced an average of 55 μm reduction in CFT and 0.2 log MAR gain in visual acuity at 12 months (P<0.001). No patient required rescue anti-VEGF injections. Group 2 (n=30) experienced no significant change in CFT or visual acuity by 6 months despite retreatment with STMP in 19 eyes. From 6 to 12 months follow-up, all the patients in group 2 received rescue Bevacizumab injections that resulted in 307 μm reduction in CFT and 0.3 log MAR improvement in visual acuity (P<0.001). No adverse effects from STMP laser were recorded. Severity of edema can influence the effects of STMP laser. STMP monotherapy is safe and effective in treating edema of mild to moderate severity.


Proteomic analysis of exosomes from human neural stem cells by flow field-flow fractionation and nanoflow liquid chromatography-tandem mass spectrometry.
Exosomes, small membrane vesicles secreted by a multitude of cell types, are involved in a wide range of physiological roles such as intercellular communication, membrane exchange between cells, and degradation as an alternative to lysosomes. Because of the small size of exosomes (30-100 nm) and the limitations of common separation procedures including ultracentrifugation and flow cytometry, size-based fractionation of exosomes has been challenging. In this study, we used flow field-flow fractionation (FlFFF) to fractionate exosomes according to differences in hydrodynamic diameter. The exosome fractions collected from FlFFF runs were examined by transmission electron microscopy (TEM) to morphologically confirm their identification as exosomes. Exosomal lysates of each fraction were digested and analyzed using nanoflow LC-ESI-MS-MS for protein identification. FIFFF, coupled with mass spectrometry, allows nanoscale size-based fractionation of exosomes and is more applicable to primary cells and stem cells since it requires much less starting material than conventional gel-based separation, in-gel digestion and the MS-MS method.


Fractional clearance of urate: validation of measurement in spot-urine samples in healthy subjects and gouty patients.
Hyperuricemia is the greatest risk factor for gout and is caused by an overproduction and/or inefficient renal clearance of urate. The fractional renal clearance of urate (FCU, renal clearance of urate/renal clearance of creatinine) has been proposed as a tool to identify subjects who manifest inefficient clearance of urate. The aim of the present studies was to validate the measurement of FCU by using spot-urine samples as a reliable indicator of the efficiency of the kidney to remove urate and to explore its distribution in healthy subjects and gouty patients. Timed (spot, 2-hour, 4-hour, 6-hour, 12-hour, and 24-hour) urine collections were used to derive FCU in 12 healthy subjects. FCUs from spot-urine samples were then determined in 13 healthy subjects twice a day, repeated on 3 nonconsecutive days. The effect of allopurinol, probenecid, and the combination on FCU was explored in 11 healthy subjects. FCU was determined in 36 patients with gout being treated with allopurinol. The distribution of FCU was examined in 118 healthy subjects and compared with that from the 36 patients with gout. No substantive or statistically significant differences were observed between the FCUs derived from spot and 24-hour urine collections. Coefficients of variation (CVs) were both 28%. No significant variation in the spot FCU was obtained either within or between days, with mean intrasubject CV of 16.4%. FCU increased with probenecid (P < 0.05), whereas allopurinol did not change the FCU in healthy or gouty subjects. FCUs of patients with gout were lower than the FCUs of healthy subjects (4.8% versus 6.9%; P < 0.0001). The present studies indicate that the spot-FCU is a convenient, valid, and reliable indicator of the efficiency of the kidney in removing urate from the blood and thus from tissues. Spot-FCU determinations may provide useful correlates in studies investigating molecular mechanisms underpinning the observed range of efficiencies of the kidneys in clearing urate from the blood. ACTRN12611000743965.


Structure, dynamics and functions of UBQLNs: at the crossroads of protein quality control machinery.
Cells rely on protein homeostasis to maintain proper biological functions. Dysregulation of protein homeostasis contributes to the pathogenesis of many neurodegenerative diseases and cancers. Ubiquilins (UBQLNs) are versatile proteins that engage with many components of protein quality control (PQC) machinery in cells. Disease-linked mutations of UBQLNs are most commonly associated with amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), and other neurodegenerative disorders. UBQLNs play well-established roles in PQC processes, including facilitating degradation of substrates through the ubiquitin-proteasome system (UPS), autophagy, and endoplasmic-reticulum-associated protein degradation (ERAD) pathways. In addition, UBQLNs engage with chaperones to sequester, degrade, or assist repair of misfolded client proteins. Furthermore, UBQLNs regulate DNA damage repair mechanisms, interact with RNA-binding proteins (RBPs), and engage with cytoskeletal elements to regulate cell differentiation and development. Important to the myriad functions of UBQLNs are its multidomain architecture and ability to self-associate. UBQLNs are linked to numerous types of cellular puncta, including stress-induced biomolecular condensates, autophagosomes, aggresomes, and aggregates. In this review, we focus on deciphering how UBQLNs function on a molecular level. We examine the properties of oligomerization-driven interactions among the structured and intrinsically disordered segments of UBQLNs. These interactions, together with the knowledge from studies of disease-linked mutations, provide significant insights to UBQLN structure, dynamics and function.


The sentinel hyperplastic polyp: a marker for synchronous neoplasia in the proximal colon.
We prospectively screened 129 asymptomatic subjects (mean age 64 yr) with flexible sigmoidoscopy. Colonoscopy was performed at a later date, regardless of the sigmoidoscopic result. Our intent was 1) to establish the prevalence of proximal neoplasms in patients with and without hyperplastic polyps within reach of the 60-cm sigmoidoscope and 2) to determine whether a distal (sentinel) hyperplastic polyp predicts the presence of synchronous neoplastic polyps higher up in the colon. Our results show that 15% of asymptomatic adult subjects without polyps on sigmoidoscopy have adenomas in proximal colonic segments that can be diagnosed only by colonoscopy. By comparison, proximal neoplasms were detected in 32% (p less than 0.05) and 37% (p less than 0.05) of patients when hyperplastic or adenomatous polyps, respectively, were present on the sigmoidoscopic examination. This finding suggests that a distal (sentinel) hyperplastic polyp by itself may be a marker for neoplastic polyps in proximal colonic segments. Also, the "index" adenoma and "sentinel" hyperplastic polyp may be equivalent for predicting the presence of proximal neoplasms. The observed detection rates for these polyps were both significantly higher than expected when compared to patients who did not have polyps in the distal colon or rectum. If these results can be confirmed by a larger prospective trial, then full colonoscopy for detection of proximal neoplasms may be indicated when either an index adenoma or sentinel hyperplastic polyp is detected by sigmoidoscopy.


Gratuitous mRNA localization in the Drosophila oocyte.
Many of the genes that control pattern formation in Drosophila encode mRNAs that are localized to discrete regions of the oocyte during oogenesis. While such localization is generally assumed to be important for the pattern-forming activities of these genes, this has been rigorously demonstrated in only a few cases. Here we address the role of mRNA localization for the dorsoventral patterning gene K10. K10 mRNA is localized to the oocyte's anterior cortex following its transport into the cell during early stages of oogenesis. We show that mutations in cappuccino and spire, which permit K10 mRNA transport, but prevent subsequent anterior localization, do not disrupt the synthesis or localization of K10 protein. We also show that modified K10 transgenes that produce transcripts which are uniformly distributed throughout the oocyte, or which are mislocalized to the oocyte's posterior pole, produce localized and functional K10 protein. We conclude that the anterior localization of K10 mRNA is not important for K10 protein targeting or gene function. We propose that the anterior localization of K10, and probably other mRNAs, is a by-product of mRNA transport and does not necessarily reflect a requirement for localization per se.


Persistent left superior vena cava: Review of embryologic anatomy and considerations for cardiopulmonary bypass.
The purpose of this report is to alert the perfusionist to the anatomy of persistent left superior vena cava (PLSVC) and the intraoperative complications that this anomaly may present. The perfusionist should be able to devise a venous circuit that will accommodate three cannulae. He should also be aware that, during cardiopulmonary bypass, a copious volume of deoxygenated blood from the left heart sump, with simultaneous cerebral congestion, may signal the presence of undocumented PLSVC.


Management of malunion of the proximal humerus: current concepts.
Proximal humerus fractures remain one of the most common orthopaedic injuries, particularly in the elderly. Displaced fractures often require surgery, and management can be challenging because of comminution and poor bone quality. Despite advances in surgical technique and implant design, reoperation for malunion or nonunion of the tuberosity (arthroplasty) or screw penetration (open reduction and internal fixation) remains problematic. Recent studies have demonstrated acceptable results following nonsurgical management of displaced proximal humerus fractures in elderly, low-demand patients. In younger, more active patients, reduced function and pain that accompany select proximal humeral malunions are generally poorly tolerated. Surgical options for symptomatic, malunited tuberosities include osteotomy, tuberoplasty with rotator cuff repair and subacromial decompression, or decompression alone. Surgical neck malunion can be managed with corrective osteotomy and preservation of the native joint. Arthroplasty is reserved for complex malunions with joint incongruity. Surgical management of symptomatic proximal humeral malunion remains challenging, but good outcomes can be achieved with proper patient selection.


Coherent thermal emission by excitation of magnetic polaritons between periodic strips and a metallic film.
The present paper theoretically demonstrates coherent thermal emission in the infrared region by exciting magnetic polaritons between metallic gratings and an opaque metallic film, separated by a dielectric spacer. The coupling of the metallic strips and the film induces a magnetic response that is characterized by a negative permeability and positive permittivity. On the other hand, the metallic film intrinsically exhibits a negative permittivity and positive permeability in the near infrared. This artificial structure is equivalent to a pair of single-negative materials. By exciting surface magnetic polaritons, large emissivity peaks can be achieved at the resonance frequencies and are almost independent of the emission angle. The resonance frequency of the magnetic response can be predicted by an analogy to an inductor and capacitor circuit. The proposed structure can be easily constructed using micro/nanofabrication.


Validation of web-based, multiple 24-h recalls combined with nutritional supplement intake questionnaires against nitrogen excretions to determine protein intake in Dutch elite athletes.
Information on dietary composition is vitally important for elite athletes to optimise their performance and recovery, which requires valid tools. The aim of the present study was to investigate the validity of assessing protein intake using three web-based 24-h recalls and questionnaires, by comparing these with three urinary N excretions on the same day. A total of forty-seven Dutch elite top athletes, both disabled and non-disabled, aged between 18 and 35 years, with a BMI of 17·5-31 kg/m2, exercising >12 h/week were recruited. Estimated mean dietary protein intake was 109·6 (sd 33·0) g/d by recalls and questionnaires v. 141·3 (sd 38·2) g/d based on N excretions in urine; the difference was 25·5 (sd 21·3) % between the methods (P<0·05). We found a reasonably good association between methods for protein intake of 0·65 (95 % CI 0·45, 0·79). On an individual level, under-reporting was larger with higher protein intakes than with lower intakes. No significant differences were found in reporting absolute differences between subcategories (sex, under-reporting, BMI, collection of recalls within a certain amount of time and using protein supplements or not). In conclusion, combined, multiple, 24-h recalls and questionnaires underestimated protein intake in these young elite athletes more than that reported for non-athlete populations. The method proved to be suitable for ranking athletes according to their protein intake as needed in epidemiological studies. On an individual level, the magnitude of underestimation was about equal for all athletes except for those with very high protein intakes.


Multi-ancestry genome-wide gene-smoking interaction study of 387,272 individuals identifies new loci associated with serum lipids.
The concentrations of high- and low-density-lipoprotein cholesterol and triglycerides are influenced by smoking, but it is unknown whether genetic associations with lipids may be modified by smoking. We conducted a multi-ancestry genome-wide gene-smoking interaction study in 133,805 individuals with follow-up in an additional 253,467 individuals. Combined meta-analyses identified 13 new loci associated with lipids, some of which were detected only because association differed by smoking status. Additionally, we demonstrate the importance of including diverse populations, particularly in studies of interactions with lifestyle factors, where genomic and lifestyle differences by ancestry may contribute to novel findings.


Tuberculosis verrucosa cutis in a patient with keloid over ear lobule.
Tuberculosis (TB) is a global health problem with predominance in resource-poor countries. Extrapulmonary TB constitutes approximately 10% of the cases and can present as different forms depending on the route of entry. Cutaneous TB is a small subset of the extrapulmonary TB. Cutaneous TB can either be of primary origin or secondary origin or it can be a part of a systemic infection. TB verrucosa cutis (TBVC) is the exogenous reinfection of TB in a previously sensitised patient. Routine cultures do not have a high yield in TBVC due to the paucibacillary nature of the organism. Here we are presenting a rare case of a patient who developed in a keloid over the right ear lobule following ear piercing. This has only been presented once in the previous literature and for the first time in the head and neck region.


Paravertebral foramen screw fixation for posterior cervical spine fusion: biomechanical study and description of a novel technique.
OBJECTIVE Fusion surgery with instrumentation is a widely accepted treatment for cervical spine pathologies. The authors propose a novel technique for subaxial cervical fusion surgery using paravertebral foramen screws (PVFS). The authors consider that PVFS have equal or greater biomechanical strength than lateral mass screws (LMS). The authors' goals of this study were to conduct a biomechanical study of PVFS, to investigate the suitability of PVFS as salvage fixation for failed LMS, and to describe this novel technique. METHODS The authors harvested 24 human cervical spine vertebrae (C3-6) from 6 fresh-frozen cadaver specimens from donors whose mean age was 84.3 ± 10.4 years at death. For each vertebra, one side was chosen randomly for PVFS and the other for LMS. For PVFS, a 3.2-mm drill with a stopper was advanced under lateral fluoroscopic imaging. The drill stopper was set to 12 mm, which was considered sufficiently short not to breach the transverse foramen. The drill was directed from 20° to 25° medially so that the screw could purchase the relatively hard cancellous bone around the entry zone of the pedicle. The hole was tapped and a 4.5-mm-diameter × 12-mm screw was inserted. For LMS, 3.5-mm-diameter × 14-mm screws were inserted into the lateral mass of C3-6. The pullout strength of each screw was measured. After pullout testing of LMS, a drill was inserted into the screw hole and the superior cortex of the lateral mass was pried to cause a fracture through the screw hole, simulating intraoperative fracture of the lateral mass. After the procedure, PVFS for salvage (sPVFS) were inserted on the same side and pullout strength was measured. RESULTS The CT scans obtained after screw insertion revealed no sign of pedicle breaching, violation of the transverse foramen, or fracture of the lateral mass. A total of 69 screws were tested (23 PVFS, 23 LMS, and 23 sPVFS). One vertebra was not used because of a fracture that occurred while the specimen was prepared. The mean bone mineral density of the specimens was 0.29 ± 0.10 g/cm3. The mean pullout strength was 234 ± 114 N for PVFS, 158 ± 91 N for LMS, and 195 ± 125 N for sPVFS. The pullout strength for PVFS tended to be greater than that for LMS. However, the difference was not quite significant (p = 0.06). CONCLUSIONS The authors introduce a novel fixation technique for the subaxial cervical spine. This study suggests that PVFS tend to provide stronger fixation than LMS for initial applications and fixation equal to LMS for salvage applications. If placement of LMS fails, PVFS can serve as a salvage fixation technique.


The effects of margination and red cell augmented platelet diffusivity on platelet adhesion in complex flow.
The effects of platelet margination and enhanced platelet diffusivity, as induced by red blood cells, on the adhesion of platelets, were investigated for a range of haematocrits, under 2D axi-symmetric flow, simulating previous in vitro experiments [Microvasc. Res. 17 (1979), 238-262]. The effect of margination was incorporated via use of an elevated platelet inlet mass fraction, Phie, in a manner similar to that of Wootton [Ann. Biomed. Eng. 29 (2001), 321-329], and a shear and haematocrit dependent platelet diffusivity, according to the model presented by Zydney and Colton [Physico Chem. Hydrodyn. 10 (1988), 77-96] was used. A combination of the two models was required to simulate the deposition of platelets to a collagen coated surface, under the complex flow, which exhibited a recirculation zone and stagnation point. Results obtained showed qualitative agreement with in vitro results, for a range of haematocrits (11-50%), and also showed that the effects of margination were not linearly dependent on haematocrit. Agreement may be improved in future simulations by incorporating the effects of depleted cell concentrations in the vortex which have been observed previously [Phil. Trans. Roy. Soc. (Lond.) B279 (1977), 413-445]. It would also be advantageous to devise a full mathematical description for platelet margination effects as a function of shear rate and haematocrit and a description of the accompanying effect of apparent blood viscosity.


Identification of Ideal Allele Combinations for the Adaptation of Spring Barley to Northern Latitudes.
The northwards expansion of barley production requires adaptation to longer days, lower temperatures and stronger winds during the growing season. We have screened 169 lines of the current barley breeding gene pool in the Nordic region with regards to heading, maturity, height, and lodging under different environmental conditions in nineteen field trials over 3 years at eight locations in northern and central Europe. Through a genome-wide association scan we have linked phenotypic differences observed in multi-environment field trials (MET) to single nucleotide polymorphisms (SNP). We have identified an allele combination, only occurring among a few Icelandic lines, that affects heat sum to maturity and requires 214 growing degree days (GDD) less heat sum to maturity than the most common allele combination in the Nordic spring barley gene pool. This allele combination is beneficial in a cold environment, where autumn frost can destroy a late maturing harvest. Despite decades of intense breeding efforts relying heavily on the same germplasm, our results show that there still exists considerable variation within the current breeding gene pool and we identify ideal allele combinations for regional adaptation, which can facilitate the expansion of cereal cultivation even further northwards.


Hippocampal HDAC4 contributes to postnatal fluoxetine-evoked depression-like behavior.
Fluoxetine treatment in adulthood evokes antidepressant and anxiolytic responses. Paradoxically, postnatal fluoxetine (PNFlx) induces persistent depression- and anxiety-like behaviors. The mechanistic underpinnings of this paradox remain poorly understood. Here, we examined specific molecular changes in the rat hippocampus that accompany perturbed emotionality observed across life following PNFlx. PNFlx-induced hippocampal gene regulation observed in microarray and quantitative PCR studies indicate functional enrichment of genes involved in response to organic substances, protein kinase pathways, DNA binding, and transcriptional repression. We noted specific transcripts (Hdac4, mammalian target of rapamycin (mTOR), Gnai1, protein kinase C gamma (Prkcc), and hyperpolarization-activated cyclic nucleotide-gated channel 1 (Hcn1)) that were consistently dysregulated across life, and selectively influenced by postnatal, but not adult, fluoxetine. Increased histone deacetylase-4 (HDAC4) recruitment, accompanied by decreased activating histone acetylation marks at the mTOR and Gnai1 promoters, indicate a role for HDAC4 in PNFlx-mediated gene dysregulation. Strikingly, coadministration of the HDAC inhibitor sodium butyrate with PNFlx prevented the dysregulation of Hdac4 and mTOR, and the emergence of depression- and anxiety-like behavior. Importantly, we also find that retreatment of PNFlx animals with fluoxetine in adulthood reversed the increased Hdac4 expression, prevented HDAC4 recruitment to the mTOR and Gnai1 promoters, and attenuated the decline in mTOR and Gnai1 expression, coincident with normalization of PNFlx-evoked depression- and anxiety-like behavior. Further, we show that viral-mediated hippocampal overexpression of Hdac4 was sufficient to induce depression-, but not anxiety-, like behavior in adulthood. Our results highlight the unique nature of molecular signatures evoked by PNFlx, and implicate HDAC4 in the dysregulated gene expression and emergence of perturbed emotionality following fluoxetine exposure in early life.


Impact of disasters on child stunting in Nepal.
Stunting is a major public health problem that results from inadequate nutritional intake over a long period of time. Disasters have major implications in poor and vulnerable children. The aim of this study was, therefore, to assess the impact of disasters on child stunting in Nepal. A sample consisting of 2,111 children aged 6-59 months was obtained from the 2011 Nepal Demographic and Health Survey. We used bivariate and multivariate analyses to examine moderate and severe stunting against disaster, controlling for all possible confounders. Out of the total study sample, 43% were stunted (17.1% severely and 25.9% moderately). The final model, after adjusting for confounders, showed that epidemics have no impact on child stunting (adjusted odds ratio [OR] =1.14, 95% confidence interval [CI]: 0.66, 1.97 and adjusted OR =1.04, 95% CI: 0.66, 1.65 for severe and moderate stunting, respectively). Floods have impact on child stunting (adjusted OR =0.57, 95% CI: 0.31, 0.96 and adjusted OR =0.66, 95% CI: 0.41, 0.94 for severe and moderate stunting, respectively). However, children aged 6-11 months, nonvaccinated children, children of working women, children who live in mountainous areas, and children from the poorest households were more likely to be moderately stunted. Similarly, children aged 36-47 months, Dalit and other ethnic groups, children from rural settings, and children from the poorest households were more likely to be severely stunted. This article illustrates the need to rethink about child stunting in Nepal. This study suggests need for further research, integration of disaster data in the Nepal Demography Health Survey, educational interventions, public awareness, promotion of vaccination, and equity in health service delivery.


The complex and intriguing lives of PIP2 with ion channels and transporters.
Phosphatidylinositol-4,5-bisphosphate (PIP(2)), the precursor of several signaling molecules in eukayotic cells, is itself also used by cells to signal to membrane-associated proteins. PIP(2) anchors numerous signaling molecules and cytoskeleton at the cell membrane, and the metabolism of PIP(2) is closely connected to membrane trafficking. Recently, ion transporters and channels have been discovered to be regulated by PIP(2). Systems reported to be activated by PIP(2) include (i) plasmalemmal calcium pumps (PMCA), (ii) cardiac sodium-calcium exchangers (NCX1), (iii) sodium-proton exchangers (NHE1-4), (iv) a sodium-magnesium exchanger of unknown identity, (v) all inward rectifier potassium channels (KATP, IRK, GIRK, and ROMK channels), (vi) epithelial sodium channels (ENaC), and (vii) ryanodine-sensitive calcium release channels (RyR). Systems reported to be inhibited by PIP(2) include (i) cyclic nucleotide-gated channels of the rod (CNG), (ii) transient receptor potential-like (TRPL) Drosophila phototransduction channels, (iii) capsaicin-activated transient receptor potential (TRP) channels (VR1), and (iv) IP(3)-gated calcium release channels (IP3R). Systems that appear to be completely insensitive to PIP(2) include (i) voltage-gated sodium channels, (ii) most voltage-gated potassium channels, (iii) sodium-potassium pumps, (iv) several neurotransmitter transporters, and (v) cystic fibrosis transmembrane receptor (CFTR)-type chloride channels. Presumably, local changes of the concentration of PIP(2) in the plasma membrane represent cell signals to those mechanisms sensitive to PIP(2) changes. Unfortunately, our understanding of how local PIP(2) concentrations are regulated remains very limited. One important complexity is the probable existence of phospholipid microdomains, or lipid rafts. Such domains may serve to localize PIP(2) and thereby PIP(2) signaling, as well as to organize PIP(2) binding partners into signaling complexes. A related biological role of PIP(2) may be to control the activity of ion transporters and channels during biosynthesis or vesicle trafficking. Low PIP(2) concentrations in the secretory pathway would inactivate all of the systems that are stimulated by PIP(2). How, in detail, is PIP(2) used by cells to control ion channel and transporter activities? Further progress requires an improved understanding of lipid kinases and phosphatases, how they are regulated, where they are localized in cells, and with which ion channels and transporters they might localize.


Representational accuracy of stochastic neural populations.
Fisher information is used to analyze the accuracy with which a neural population encodes D stimulus features. It turns out that the form of response variability has a major impact on the encoding capacity and therefore plays an important role in the selection of an appropriate neural model. In particular, in the presence of baseline firing, the reconstruction error rapidly increases with D in the case of Poissonian noise but not for additive noise. The existence of limited-range correlations of the type found in cortical tissue yields a saturation of the Fisher information content as a function of the population size only for an additive noise model. We also show that random variability in the correlation coefficient within a neural population, as found empirically, considerably improves the average encoding quality. Finally, the representational accuracy of populations with inhomogeneous tuning properties, either with variability in the tuning widths or fragmented into specialized subpopulations, is superior to the case of identical and radially symmetric tuning curves usually considered in the literature.


Gender differences in responses to moral dilemmas: a process dissociation analysis.
The principle of deontology states that the morality of an action depends on its consistency with moral norms; the principle of utilitarianism implies that the morality of an action depends on its consequences. Previous research suggests that deontological judgments are shaped by affective processes, whereas utilitarian judgments are guided by cognitive processes. The current research used process dissociation (PD) to independently assess deontological and utilitarian inclinations in women and men. A meta-analytic re-analysis of 40 studies with 6,100 participants indicated that men showed a stronger preference for utilitarian over deontological judgments than women when the two principles implied conflicting decisions (d = 0.52). PD further revealed that women exhibited stronger deontological inclinations than men (d = 0.57), while men exhibited only slightly stronger utilitarian inclinations than women (d = 0.10). The findings suggest that gender differences in moral dilemma judgments are due to differences in affective responses to harm rather than cognitive evaluations of outcomes.


The microbial metabolites, short-chain fatty acids, regulate colonic Treg cell homeostasis.
Regulatory T cells (Tregs) that express the transcription factor Foxp3 are critical for regulating intestinal inflammation. Candidate microbe approaches have identified bacterial species and strain-specific molecules that can affect intestinal immune responses, including species that modulate Treg responses. Because neither all humans nor mice harbor the same bacterial strains, we posited that more prevalent factors exist that regulate the number and function of colonic Tregs. We determined that short-chain fatty acids, gut microbiota-derived bacterial fermentation products, regulate the size and function of the colonic Treg pool and protect against colitis in a Ffar2-dependent manner in mice. Our study reveals that a class of abundant microbial metabolites underlies adaptive immune microbiota coadaptation and promotes colonic homeostasis and health.


The Social Value of Health Research and the Worst Off.
In this article we argue that the social value of health research should be conceptualized as a function of both the expected benefits of the research and the priority that the beneficiaries deserve. People deserve greater priority the worse off they are. This conception of social value can be applied for at least two important purposes: (1) in health research priority setting when research funders, policy-makers, or researchers decide between alternative research projects; and (2) in evaluating the ethics of proposed research proposals when research ethics committees (RECs) assess whether the social value of the research is sufficient to justify the risks and burdens to research participants and others. In assessing how far a proposed research project will advance the interests of people who are more disadvantaged, research priority setters and RECs should examine (at least) the diseases that the research targets and the type of research. Just as certain diseases impose a greater burden on people who are more disadvantaged, so certain types of intervention and forms of research are more likely to benefit people who are more disadvantaged. We outline which populations are likely to be representative of the global worst off and identify what types of health research, and which disease categories, are priorities for these populations.


Late toxicity rates following definitive radiotherapy for prostate cancer.
Most patients survive many years following external beam radiotherapy (RT) for nonmetastatic prostate cancer and are therefore at risk for late treatment sequelae. The relationships between RT dose, treatment technique, and late toxicity rates are incompletely understood. Here we perform a meta-analysis and systematic review to characterize those effects. We performed a review of published series that report late gastrointestinal (GI) and genitourinary (GU) toxicity rates following definitive RT for prostate cancer using the RTOG Late Radiation Morbidity Scoring Schema. Univariate analyses were performed to test RT technique, RT dose, pelvic irradiation, and androgen deprivation therapy (ADT) as predictors of moderate (grade ≥ 2) and severe (grade ≥ 3) GI and GU toxicity. To isolate the effect of radiotherapy dose on late toxicity, we also performed a meta-analysis restricted to randomized trials that tested RT dose escalation. Statistical analyses were repeated using the subset of studies that utilized escalated RT doses. Twenty published reports detailing the treatment techniques and toxicity outcomes of 35 patient series including a total of 11,835 patients were included in this analysis. Median rates of moderate late toxicity were 15% (GI) and 17% (GU). For severe effects, these values were 2% (GI) and 3% (GU). Meta-analysis of five randomized trials revealed that an 8-10 Gy increase in RT dose increases the rate of both moderate (OR = 1.63, 95% CI: [1.44 to 1.82], p < 0.001) and severe (OR = 2.03, 95% CI: [1.64 to 2.42], p < 0.001) late GI toxicity. Among 17 series where doses of at least 74 Gy were utilized, use of intensity-modulated radiotherapy (IMRT) or proton beam radiotherapy (PBRT) was associated with a significant decrease in the reported rate of severe GI toxicity compared to 3-D RT. Meta-analysis of randomized dose escalation trials demonstrates that late toxicity rates increase with RT dose. Series where dose escalated RT is delivered using IMRT or PBRT have relatively short follow up but report lower late GI toxicity rates than those employing 3-D RT.


Allometric equations for integrating remote sensing imagery into forest monitoring programmes.
Remote sensing is revolutionizing the way we study forests, and recent technological advances mean we are now able - for the first time - to identify and measure the crown dimensions of individual trees from airborne imagery. Yet to make full use of these data for quantifying forest carbon stocks and dynamics, a new generation of allometric tools which have tree height and crown size at their centre are needed. Here, we compile a global database of 108753 trees for which stem diameter, height and crown diameter have all been measured, including 2395 trees harvested to measure aboveground biomass. Using this database, we develop general allometric models for estimating both the diameter and aboveground biomass of trees from attributes which can be remotely sensed - specifically height and crown diameter. We show that tree height and crown diameter jointly quantify the aboveground biomass of individual trees and find that a single equation predicts stem diameter from these two variables across the world's forests. These new allometric models provide an intuitive way of integrating remote sensing imagery into large-scale forest monitoring programmes and will be of key importance for parameterizing the next generation of dynamic vegetation models.


The stromal cell antigen CD248 (endosialin) is expressed on naive CD8+ human T cells and regulates proliferation.
CD248 (endosialin) is a transmembrane glycoprotein that is dynamically expressed on pericytes and fibroblasts during tissue development, tumour neovascularization and inflammation. Its role in tissue remodelling is associated with increased stromal cell proliferation and migration. We show that CD248 is also uniquely expressed by human, but not mouse (C57BL/6), CD8(+) naive T cells. CD248 is found only on CD8(+) CCR7(+) CD11a(low) naive T cells and on CD8 single-positive T cells in the thymus. Transfection of the CD248 negative T-cell line MOLT-4 with CD248 cDNA surprisingly reduced cell proliferation. Knock-down of CD248 on naive CD8 T cells increased cell proliferation. These data demonstrate opposing functions for CD248 on haematopoietic (CD8(+)) versus stromal cells and suggests that CD248 helps to maintain naive CD8(+) human T cells in a quiescent state.


Spotting sporotrichosis skin infection: The first Australian paediatric case series.
Sporotrichosis is a dermatomycosis caused by the dimorphic fungus, Sporothrix schenckii, with various outbreaks across Australia attributed to mouldy hay. Our objective was to investigate the clinical presentation and management of cutaneous sporotrichosis in a paediatric population of Western Australia. A retrospective case review was performed for S. schenckii infections in children below 18 years, between January 2000 and November 2017. Cases were identified from the state-wide laboratory database and additional clinical data obtained from medical records. Thirty-two cases of microbiologically proven S. schenckii infection were identified, mostly from rural areas (n = 20, 63%). Complete clinical data were available for 11 cases (34%). The most common risk factors were exposure to farm animals and hay, arthropod bites and outdoor activities. The median duration from symptom onset to correct diagnosis was 6 weeks (interquartile range: 4-7 weeks). Most cases were initially treated with multiple, broad-spectrum antibacterial agents (n = 7, 64%). Targeted therapy (itraconazole) was used in all cases once the diagnosis was made, with a median treatment duration of 5 months (interquartile range: 4-6 months). Morbidity included scarring (n = 4, 31%), itraconazole associated diarrhoea (n = 1, 8%) and mild hepatotoxicity (n = 1, 8%). Summarising the clinical experience of these cases is a useful guide for clinical recognition and may serve to shorten the interval between onset and diagnosis, and avoid the need for antibacterial therapy. These data highlight the importance of recognising Sporotrichosis in children outside an outbreak setting, leading to timely diagnosis and appropriate treatment with antifungal agents.


Genetic heterogeneity in ten families with myoclonus-dystonia.
Myoclonus-dystonia (M-D) is a movement disorder with autosomal dominant inheritance and reduced penetrance but may also occur sporadically. Recently, mutations in the epsilon-sarcoglycan gene (SGCE) were shown to cause M-D. Furthermore, single variants in the dopamine D2 receptor (DRD2) and DYT1 genes were found in combination with SGCE mutations in two M-D families, and another M-D locus was recently mapped to chromosome 18p11 in one family. The authors clinically and genetically characterised ten consecutive cases with myoclonus-dystonia; seven familial and three sporadic. Twenty nine M-D patients and 40 unaffected family members underwent a standardised clinical examination by a movement disorder specialist. Index cases were screened for mutations in the SGCE, DYT1, and DRD2 genes and for deletions of the SGCE gene. Suitable mutation negative families were tested for linkage to the SGCE region and to chromosome 18p11. Two SGCE mutations were detected among the seven familial but no mutation in the sporadic cases. Haplotype analysis at the new M-D locus was compatible with linkage in two families and excluded in another family, suggesting at least one additional M-D gene. There were no obvious clinical differences between M-D families with and without detected mutations. M-D is genetically heterogeneous with SGCE mutations accounting for the disease in only part of the clinically typical cases.


Effects of global intermittency on laser propagation in the atmosphere.
The statistical description of atmospheric turbulence and laser propagation through the atmosphere is complicated by global intermittency, the slow variations in space and time of the basic turbulence parameters. The effects of global intermittency of atmospheric turbulence on the mutual coherence function and the irradiance spectrum are investigated with numerical simulation. The effects of global intermittency are negligible when the path length is of the order of 10 correlation scales of global intermittency. This condition is readily satisfied for typical atmospheric parameters. The higher moments of irradiance appear to be more sensitive to the effects of global intermittency.


The burden of mental health-related mortality in the Baltic States in 2007-2018.
The problem of underestimating the burden of mental health-related mortality is widely discussed in the public health literature. Relevant scientific evidence from societies experiencing the largest burden of mental health mortality is important for better understanding global and national mental health challenges and improving policies. Three Baltic States - Estonia, Lithuania, and Latvia - are countries in the Central and Eastern European region that experienced post-soviet transition trauma and showed among the highest suicide and alcohol-related mortality rates in Europe. This study aimed to examine the change in the burden of mental health-related mortality in three Baltic States in the context of consistent growth in life expectancy in 2007-2018. We calculated age-standardized years of life lost due to specific mental health-related causes of death in three Baltic countries from 2007 to 2018. Four mental health-related causes of death groups were analyzed: (i) all mental and behavioural disorders; (ii) intentional self-harm; (iii) main substance use-related causes of death; and (iv) external causes of death. The number of deaths came from the WHO Mortality Database; population exposures were extracted from the Human Mortality Database. We found that the proportion of age-standardized years of life lost due to mental disorders was relatively low in all three countries. It varied from 0.2% for females in Lithuania in 2009 to 2.4% for males in Estonia in 2007. However, the proportion of age-standardized years of life lost from self-harm and substance use remained high. In 2018, the proportion of age-standardized years of life lost due to self-harm was highest among males in Lithuania (4.1%) while the highest proportion due to substance use-related causes of death was among males in Estonia (7.3%). Our findings indicate that the burden of mental health-related mortality remained high and showed divergent temporal changes across the three countries. In the context of the Baltic States and other post-soviet countries, fractions of various external causes of death and alcohol-related causes of death should be considered in assessing the total burden of mental health-related mortality.


Three-dimensional kinematics of skeletal elements in avian prokinetic and rhynchokinetic skulls determined by Roentgen stereophotogrammetry.
Several different types of cranial kinesis are present within modern birds, enabling them to move (part of) the upper bill relative to the braincase. This movement of the upper bill results from movement of the quadrate and the pterygoid-palatine complex (PPC). The taxon Palaeognathae is characterised by a very distinct PPC and a special type of cranial kinesis (central kinesis) that is very different from that found in the Neognathae. This has led some authors to hypothesise that there is a functional relationship between the morphology of the PPC and the type of cranial kinesis. This hypothesis is tested here by analysing the movement pattern of both the upper bill and the PPC in birds with three different types of cranial kinesis: prokinesis, distal rhynchokinesis and central rhynchokinesis. Movement patterns were determined using a Roentgen stereophotogrammetry method, which made it possible to detect very small displacements (0.5 mm) of bony elements in three dimensions, while the jaw muscles and ligaments remained intact. We found that in all types of kinesis investigated the movements of the quadrate, jugal bars and PPC are similar. Movement of the quadrate is transferred to the upper beak by the jugal bar and the PPC, which moves almost exclusively forwards and backwards, thereby elevating or depressing the upper bill. The differences between the types of kinesis lie only in the position of the point of rotation. These findings indicate that there is no correlation between the specific morphology of the PPC and the type of cranial kinesis. Several other factors, including the external forces applied during food acquisition, may influence the morphology of the PPC. Differences in PPC morphology therefore appear to be the result of different functional demands acting on the system simultaneously but with different strengths, depending on the species.


Isolation of avian influenza H5N1 virus from vaccinated commercial layer flock in Egypt.
Uninterrupted transmission of highly pathogenic avian influenza virus (HPAIV) H5N1 of clade 2.2.1 in Egypt since 2006 resulted in establishment of two main genetic clusters. The 2.2.1/C group where all recent human and majority of backyard origin viruses clustered together, meanwhile the majority of viruses derived from vaccinated poultry in commercial farms grouped in 126.96.36.199 clade. In the present investigation, an HPAIV H5N1 was isolated from twenty weeks old layers chickens that were vaccinated with a homologous H5N1 vaccine at 1, 7 and 16 weeks old. At twenty weeks of age, birds showed cyanosis of comb and wattle, decrease in egg production and up to 27% mortality. Examined serum samples showed low antibody titer in HI test (Log2 3.2 ± 4.2). The hemagglutinin (HA) and neuraminidase (NA) genes of the isolated virus were closely related to viruses in 2.2.1/C group isolated from poultry in live bird market (LBM) and backyards or from infected people. Conspicuous mutations in the HA and NA genes including a deletion within the receptor binding domain in the HA globular head region were observed. Despite repeated vaccination of layer chickens using a homologous H5N1 vaccine, infection with HPAIV H5N1 resulted in significant morbidity and mortality. In endemic countries like Egypt, rigorous control measures including enforcement of biosecurity, culling of infected birds and constant update of vaccine virus strains are highly required to prevent circulation of HPAIV H5N1 between backyard birds, commercial poultry, LBM and humans.


What Makes for Successful Registry Implementation: A Qualitative Comparative Analysis.
Registry implementation is an important component of successfully achieving patient-centered medical home designation and an important part of population-based health. The purpose of this study was to examine what factors are evident in the successful implementation of a registry in a selection of Colorado practices involved in quality-improvement activities. In-depth, small-group interviews occurred at 13 practices. The data were recorded, transcribed, and qualitatively analyzed to identify key themes regarding elements of successful registry implementation. Key elements were described as conditions, then calibrated and analyzed using qualitative comparative analysis (QCA). The QCA revealed several formulas to successful registry implementation. Key conditions included the importance of Resources and Leadership along with either a Quality Improvement Mindset or a Key Person driving efforts (or both). Health System membership affected the specific formula. This study is innovative in that it examines which factors and in what combination are necessary for successful implementation of a registry. The findings have implications for primary care quality-improvement efforts.


Intrinsic and synaptic long-term depression of NTS relay of nociceptin- and capsaicin-sensitive cardiopulmonary afferents hyperactivity.
The nucleus tractus solitarius (NTS) in the caudal medulla is a gateway for a variety of cardiopulmonary afferents important for homeostatic regulation and defense against airway and cardiovascular insults and is a key central target potentially mediating the response habituation to these inputs. Here, whole-cell and field population action potential recordings and infrared imaging in rat brainstem slices in vitro revealed a compartmental pain-pathway-like organization of capsaicin-facilitated vs. nocistatin-facilitated/nociceptin-suppressed neuronal clusters in an NTS region, which receives cardiopulmonary A- and C-fiber afferents with differing capsaicin sensitivities. All capsaicin-sensitive neurons and a fraction of nociceptin-sensitive neurons expressed N-methyl-D: -aspartate (NMDA) receptor-dependent synaptic long-term depression (LTD) following afferent stimulation. All neurons also expressed activity-dependent decrease of excitability (intrinsic LTD), which converted to NMDA receptor-dependent intrinsic long-term potentiation after GABA(A) receptor blockade. Thus, distinct intrinsic and synaptic LTD mechanisms in the NTS specific to the relay of A- or C-fiber afferents may underlie the response habituation to persistent afferents hyperactivity that are associated with varying physiologic challenges and cardiopulmonary derangements-including hypertension, chronic cough, asthmatic bronchoconstriction, sustained elevated lung volume in chronic obstructive pulmonary disease or in continuous positive-airway-pressure therapy for sleep apnea, metabolic acidosis, and prolonged exposure to hypoxia at high altitude.


Prediction of the coding sequences of mouse homologues of KIAA gene: III. the complete nucleotide sequences of 500 mouse KIAA-homologous cDNAs identified by screening of terminal sequences of cDNA clones randomly sampled from size-fractionated libraries.
We have conducted a human cDNA project to predict protein-coding sequences (CDSs) in large cDNAs (> 4 kb) since 1994, and the number of newly identified genes, known as KIAA genes, already exceeds 2000. The ultimate goal of this project is to clarify the physiological functions of the proteins encoded by KIAA genes. To this end, the project has recently been expanded to include isolation and characterization of mouse KIAA-counterpart genes. We herein present the entire sequences and the chromosome loci of 500 mKIAA cDNA clones and 13 novel cDNA clones that were incidentally identified during this project. The average size of the 513 cDNA sequences reached 4.3 kb and that of the deduced amino acid sequences from these cDNAs was 816 amino acid residues. By comparison of the predicted CDSs between mouse and human KIAAs, 12 mKIAA cDNA clones were assumed to be differently spliced isoforms of the human cDNA clones. The comparison of mouse and human sequences also revealed that four pairs of human KIAA cDNAs are derived from single genes. Notably, a homology search against the public database indicated that 4 out of 13 novel cDNA clones were homologous to the disease-related genes.


Epidemiology and genetic diversity of rotavirus strains in children with acute gastroenteritis in Lahore, Pakistan.
Pakistan harbors high disease burden of gastro-enteric infections with majority of these caused by rotavirus. Unfortunately, lack of proper surveillance programs and laboratory facilities have resulted in scarcity of available data on rotavirus associated disease burden and epidemiological information in the country. We investigated 1306 stool samples collected over two years (2008-2009) from hospitalized children under 5 years of age for the presence of rotavirus strains and its genotypic diversity in Lahore. The prevalence rate during 2008 and 2009 was found to be 34% (n = 447 out of 1306). No significant difference was found between different age groups positive for rotavirus (p>0.05). A subset of EIA positive samples was further screened for rotavirus RNA through RT-PCR and 44 (49.43%) samples, out of total 89 EIA positive samples, were found positive. G and P type prevalence was found as follows: G1P [4] = 3(6.81%); G1P [6] = 9(20.45%); G1P [8] = 1(2.27%); G2P [4] = 21(47.72%); G2P [8] = 1(2.27%); G9P [4] = 1(2.27%); G9P [6] = 1(2.27%) and G9P [8] = 7(15.90%). Phylogenetic analysis revealed that the VP7 and VP4 sequences clustered closely with the previously detected strains in the country as well as Belgian rotaviruses. Antigenic characterization was performed by analyzing major epitopes in the immunodominant VP7 and VP4 gene segments. Although the neutralization conferring motifs were found variable between the Pakistani strains and the two recommended vaccines strains (Rotarix™ and RotaTeq™), we validate the use of rotavirus vaccine in Pakistan based on the proven and recognized vaccine efficacy across the globe. Our findings constitute the first report on rotavirus' genotype diversity, their phylogenetic relatedness and epidemiology during the pre-vaccination era in Lahore, Pakistan and support the immediate introduction of rotavirus vaccine in the routine immunization program of the country.


Religion, Culture and Meaning-Making Coping: A Study Among Cancer Patients in Malaysia.
The present study aimed to explore the use of meaning-making coping mechanisms (existential, spiritual and religious coping) among ethnic Malay cancer patients in Malaysia and to investigate the impact of culture on their choice of coping methods. Twenty-nine participants with various kinds of cancer were interviewed. Four kinds of coping resources emerged from analyses of the interview transcripts: (1) relying on transcendent power, (2) supernatural or mystical beliefs, (3) finding oneself in relationships with others and (4) nature. In this article, the two first resources are in focus. The present findings suggest that Malay culture, which is imbued with Islamic belief, strongly influences cancer patients' coping methods and ways of looking at their experience of being cancer patients.


Mitochondrial oxidative stress promotes atrial fibrillation.
Oxidative stress has been suggested to play a role in the pathogenesis of atrial fibrillation (AF). Indeed, the prevalence of AF increases with age as does oxidative stress. However, the mechanisms linking redox state to AF are not well understood. In this study we identify a link between oxidative stress and aberrant intracellular Ca(2+) release via the type 2 ryanodine receptor (RyR2) that promotes AF. We show that RyR2 are oxidized in the atria of patients with chronic AF compared with individuals in sinus rhythm. To dissect the molecular mechanism linking RyR2 oxidation to AF we used two murine models harboring RyR2 mutations that cause intracellular Ca(2+) leak. Mice with intracellular Ca(2+) leak exhibited increased atrial RyR2 oxidation, mitochondrial dysfunction, reactive oxygen species (ROS) production and AF susceptibility. Both genetic inhibition of mitochondrial ROS production and pharmacological treatment of RyR2 leakage prevented AF. Collectively, our results indicate that alterations of RyR2 and mitochondrial ROS generation form a vicious cycle in the development of AF. Targeting this previously unrecognized mechanism could be useful in developing effective interventions to prevent and treat AF.


THE INFLUENCE OF AGE AND OF DURATION OF TREATMENT ON THE PRODUCTION AND REPAIR OF BONE LESIONS IN EXPERIMENTAL HYPERPARATHYROIDISM.
These studies have shown that the bones of guinea pigs given daily injections of parathormone from the age of 2 to 7 days to the age of 110 to 120 days, show relatively very little effect after receiving 20 units daily during the last 65 to 87 days of treatment. But it is probable that their bones underwent decalcification early in the treatment and that subsequently the parathormone, continued at the same dosage, did not maintain the effects on the bones. Healing finally occurred despite it. The bones of guinea pigs treated with intermittent injections of large doses of parathormone from the time they were 1 week old to the age of 95 to 145 days also showed relatively few changes at the end of the treatment. The injections were given at intervals of 7 to 11 days, and were stepped up from 60 units to 140 units. From our previous experience (1) we infer that the earlier injections of parathormone produced very extensive bone changes which healed in the intervals between the injections. As the guinea pigs became older the injections of parathormone did not produce as severe effects. We have found in our studies of experimental hyperparathyroidism that the bone changes after a single large dose of parathormone in young guinea pigs are soon healed. The study of a series of animals shows that healing begins at about the 48th hour after injection, and proceeds rapidly. Between the 8th and 14 days, callus may be observed at the costochondral junctions, where fractures had occurred. Now the endosteum may be lined by osteoblasts and the vessel canals by new formed bone. In adult guinea pigs extremely large single doses had little effect on the bones in 48 hours, even though the dose killed the animal. It was only when three doses pyramided over a period of 48 hours and totaling 2580 units of parathormone were given, that moderately severe bone resorption could be demonstrated in the adult. The elevation of serum calcium may be considered as one of the indices of calcium mobilization in experimental hyperparathyroidism. When the rate of calcium excretion exceeds the rate of its mobilization, or when the animal is on a low calcium diet, hypercalcemia may be absent. It is possible to raise the serum calcium of adult guinea pigs by large single doses of parathormone, but the resulting rise is not as great as in the young (2). This is confirmatory evidence of the fact that calcium is mobilized much less rapidly from the bones of old animals than from those of young ones. Collip pointed out that young normal dogs are more susceptible to parathormone (6). This observation was corroborated by Morgan and Garrison (7). We found that the same difference held also in experimental hyperparathyroidism produced in dogs by repeated doses of parathormone (8). In man, clinical experience likewise indicates the necessity of using relatively large doses of parathormone to raise the serum calcium of adults. The serum calcium of middle-aged or old adults does not rise significantly unless as much as 100 units or more of parathormone are given daily for a number of days. Charts VI and VII, in a recent paper by Merritt and Bauer (9), support our findings of the relative difficulty of obtaining a significant elevation of serum calcium in adults. If adult guinea pigs are given daily injections of parathormone which are rapidly stepped up, the animals may be killed by the ensuing acute hyperparathyroidism, only slight bone changes being produced. However, a careful avoidance of the induction of acute hyperparathyroidism by gradual stepping up of the parathormone dose permits the employment of doses continued over a long period of time that could not possibly have been tolerated otherwise. Furthermore, healing of the lesions thus produced may occur, in spite of the continuance of parathormone at this level. It seems likely that the difference in response of young and old guinea pigs to single doses of parathormone, as indicated by the bone changes, as well as by the serum calcium and phosphorus, is related to the more rapid rate of mineral metabolism in the young, actively growing animals. The calcium mobilizing effect of parathormone is most prominent in actively growing young animals, the calcium being withdrawn from the most readily available stores-the regions of most active new bone formation and most active bone reconstruction (10). In the adult animal the calcium reserves (in the formed bone) are less susceptible to the calcium mobilizing effect of parathormone. The adult guinea pig will show relatively slight bone changes even as a result of extremely large, fatal doses of parathormone. Repeated doses, as is well known, will produce, by pyramiding, greater effects than the entire amount administered at one time. In this type of experiment the young again show greater susceptibility of the bone than the adult. In time, however, some compensation takes place, and the effects of the same doses are decreased until finally healing may occur in spite of the continued treatment. Increase of the dose, however, again elicits the parathormone effects upon the bone, as well as upon the serum calcium and phosphorus, without toxic changes (1, 8). It would seem that some compensation sets in which may be overcome by increasing the dose. This compensation is especially evident in the experiments in which the parathormone doses were stepped up gradually from small amounts. In addition to the compensation observed in young and adult animals as a result of repeated injections of parathormone, we must also consider the possibility that there is a compensating mechanism in adult animals more effective than in the young. That compensation occurs is unquestionable but its nature is not clear. Apparently it is less effective during pregnancy, doses of parathormone which produce only slight bone changes in ordinary adults causing very severe lesions in advanced pregnancy (11). Parathormone has been shown to produce only one primary effect on bone, and that is decalcification. This may come about as the result of a change in the circulating tissue fluids, the salts being dissolved out of the organic matrix, and the latter disappearing secondarily. The process is most rapid in the vicinity of most active bone formation. The osteoblasts disappear from the surfaces of bone where dissolution is occurring, and at the same time the marrow connective tissue proliferates. Fusion of cells produces osteoclasts (12), which then proceed to remove the decalcified organic matrix, with the production of the deep lacunae of Howship. Frequently leucocytes are also observed actively phagocyting the decalcified organic matrix, and often leucocytes are observed within the osteoclasts (12). Healing is associated with the complete reversal of the process. The osteoclasts disappear, the connective tissue diminishes, osteoblasts reappear, and bone formation is resumed. As we have previously stated (13), parathormone produces a more continuous effect than experimental acidosis and greater changes than are seen in experimental osteoporosis. A pronounced decalcification results from it which, with its sequelae, simulates von Recklinghausen's disease. The emphasis which the older pathologists laid on osteoclasts as a special feature of ostitis fibrosa cystica is justified, for in the experimental condition the appearance of great numbers of osteoclasts is a constant feature, whenever decalcification occurs (13). There seems to be no doubt that the giant cell tumors found in ostitis fibrosa cystica are expressions of the same pathological response. The other features of the bone changes of hyperparathyroidism-marrow hemorrhage, cysts, fractures, and osteoid proliferation-are secondary to the primary decalcification. Progress of the pathological changes leads to circulatory stasis and cyst formation. Stresses and strains exerted on the progressively weakening bone may result in microscopical or gross fractures. Osteoid tissue is, as we have previously pointed out (13), merely reparative in nature, being laid down as support to the weakened or fractured bone, or as a part of healing. Osteoid borders appear on bone surfaces 48 hours after one large dose of parathormone. The mosaic picture which we have observed in the bones of some of our animals is produced by short and irregularly disposed cement lines resulting from rapid bone transformation. Schmorl (14) recently emphasized the mosaic-like appearance of the newly formed lamellar bone in Paget's disease (ostitis fibrosa deformans). The mosaic-like appearance of bone has also been described in local bone conditions, as e.g. syphilitic periostitis, and in bone in the vicinity of cysts and giant cell tumors in von Recklinghausen's disease (ostitis fibrosa cystica). However, Schmorl claims that in no disease is the mosaic appearance so constant and the arrangement of the cement lines so irregular as in Paget's disease. In chronic experimental hyperparathyroidism (von Recklinghausen's disease), the mosaic structure is not a prominent feature because of the progressive decalcification. But the bones of our young guinea pigs which received intermittent injections showed a mosaic-like appearance indicative of the periodic decalcifications and restorations which they had undergone.