Experiences of stigma, discrimination, care and support among people living with HIV: a four country study.
While it is widely agreed that HIV-related stigma may impede access to treatment and support, there is little evidence describing who is most likely to experience different forms of stigma and discrimination and how these affect disclosure and access to care. This study examined experiences of interpersonal discrimination, internalized stigma, and discrimination at health care facilities among HIV-positive adults aged 18 years and older utilizing health facilities in four countries in Sub-Saharan Africa (N = 536). Prevalence of interpersonal discrimination across all countries was 34.6 %, with women significantly more likely to experience interpersonal discrimination than men. Prevalences of internalized stigma varied across countries, ranging from 9.6 % (Malawi) to 45.0 % (Burkina Faso). Prevalence of health care discrimination was 10.4 % across all countries. In multiple regression analyses, we found positive, significant, and independent associations between disclosure and interpersonal discrimination and disclosure and support group utilization, and positive associations between both internalized stigma and health care discrimination and referral for medications.


A sinusoidal family of unitary transforms.
A new family of unitary transforms is introduced. It is shown that the well-known discrete Fourier, cosine, sine, and the Karhunen-Loeve (KL) (for first-order stationary Markov processes) transforms are members of this family. All the member transforms of this family are sinusoidal sequences that are asymptotically equivalent. For finite-length data, these transforms provide different approximations to the KL transform of the said data. From the theory of these transforms some well-known facts about orthogonal transforms are easily explained and some widely misunderstood concepts are brought to light. For example, the near-optimal behavior of the even discrete cosine transform to the KL transform of first-order Markov processes is explained and, at the same time, it is shown that this transform is not always such a good (or near-optimal) approximation to the above-mentioned KL transform. It is also shown that each member of the sinusoidal family is the KL transform of a unique, first-order, non-stationary (in general), Markov process. Asymptotic equivalence and other interesting properties of these transforms can be studied by analyzing the underlying Markov processes.


A double-negative feedback loop between NtrBC and a small RNA rewires nitrogen metabolism in legume symbionts.
The nitrogen (N) status transduced via the NtrBC two-component system is a major signaling cue in the root nodule endosymbiosis of diazotrophic rhizobia with legumes. NtrBC is upregulated in the N-limiting rhizosphere environment at the onset of nodulation but silenced in nodules to favor the assimilation of the fixed N into plant biomass. We reported that the trans-acting sRNA NfeR1 (Nodule Formation Efficiency RNA) broadly influences the symbiotic performance of the α-rhizobium Sinorhizobium meliloti. Here, we show that NfeR1 is indeed an N-responsive sRNA that fine-tunes NtrBC output during the symbiotic transition. Biochemical and genetic approaches unveiled that NtrC and the LysR-type symbiotic regulator LsrB bind at distinct nearby sites in the NfeR1 promoter, acting antagonistically as repressor and activator of transcription, respectively. This complex transcriptional control specifies peak NfeR1 steady-state levels in N-starved and endosymbiotic bacteria. Furthermore, NfeR1 base pairs the translation initiation region of the histidine kinase coding mRNA ntrB, causing a decrease in both NtrB and NtrC abundance as assessed by double-plasmid genetic assays. In the context of endogenous regulation, NfeR1-mediated ntrBC silencing most likely amends the effective strength of the known operon autorepression exerted by NtrC. Accordingly, a lack of NfeR1 shifts the wild-type NtrBC output, restraining the fitness of free-living rhizobia under N stress and plant growth upon nodulation. The mixed NtrBC-NfeR1 double-negative feedback loop is thus an unprecedented adaptive network motif that helps α-rhizobia adjust N metabolism to the demands of an efficient symbiosis with legume plants. IMPORTANCE Root nodule endosymbioses between diazotrophic rhizobia and legumes provide the largest input of combined N to the biosphere, thus representing an alternative to harmful chemical fertilizers for sustainable crop production. Rhizobia have evolved intricate strategies to coordinate N assimilation for their own benefit with N2 fixation to sustain plant growth. The rhizobial N status is transduced by the NtrBC two-component system, the seemingly ubiquitous form of N signal transduction in Proteobacteria. Here, we show that the regulatory sRNA NfeR1 (nodule formation efficiency RNA) of the alfalfa symbiont Sinorhizobium meliloti is transcribed from a complex promoter repressed by NtrC in a N-dependent manner and feedback silences ntrBC by complementary base-pairing. These findings unveil a more prominent role of NtrC as a transcriptional repressor than hitherto anticipated and a novel RNA-based mechanism for NtrBC regulation. The NtrBC-NfeR1 double-negative feedback loop accurately rewires symbiotic S. meliloti N metabolism and is likely conserved in α-rhizobia.


Prospective study of esophageal botulinum toxin injection in high-risk achalasia patients.
Intrasphincteric injection of botulinum toxin has been reported as a safer treatment alternative to balloon dilation or myotomy in achalasia. We studied botulinum toxin injection in achalasia patients who are at high surgical risk because of age or concomitant medical problems. Consecutive patients who were elderly (age > 60 yr) or who had significant medical problems or both were enrolled after confirming achalasia by history, manometry, and esophageal scintigraphy. Patients underwent esophagogastroduodenoscopy, and 20 units of botulinum toxin were injected into each of four quadrants of the lower esophageal sphincter. Patients were interviewed at 1, 3, 5, and 6 months, and esophageal scintigraphy was repeated at 1 month. Sixteen patients with increased surgical risks were studied: many had serious coronary heart disease, diabetes, or obstructive lung disease. At 1 month, 12 of 16 patients had a clinical response but 5 developed recurrent symptoms within 6 months. One developed reflux, and two were found to have esophageal wall inflammation, loss of tissue planes, and mediastinal adhesions at subsequent myotomy. Intrasphincteric botulinum toxin injection may be appropriate in those achalasia patients who are elderly or have concomitant medical problems but concern persists regarding the length of the response and untoward side effects.


Effects of porous-hydroxyapatite incorporated into glass-ionomer sealants.
The purpose of the present study was to evaluate the mechanical and chemical properties of a novel glass ionomer cement for use as a pit and fissure sealant containing a porous hydroxyapatite, namely, apatite ionomer cement (AIC). Control sealant samples were used Fuji III (GIC-S). The experiment sealant samples (AIC-S) consisted of porous spherical hydroxyapatite (HApS) particles added at 28 wt% to GIC-S powder. The GIC-S and AIC-S samples were evaluated through mechanical strength measurements, scanning electron microscopy observations, energy dispersive X-ray spectroscopy analysis, fluoride ion release tests, and antibacterial tests. The flexural strength of the AIC-S was significantly higher than that of GIC-S for each period, 1 h, 24 h and 1 year. The fluoride release dose for AIC-S was consistently higher than that for GIC-S. In addition, the antibacterial properties of AIC-S were superior to those of GIC-S. The novel AIC-S may be a more suitable sealant material for pits and fissures with intact and/or infected enamel.


Patterning Networks of Grade IV Glioblastoma on Silicon Chip.
Glioblastoma (GBM) is the most aggressive high-grade brain cancer with a median survival time of <15 months. Due to GBMs fast and infiltrative growth patient prognosis is poor with recurrence after treatment common. Investigating GBMs ability to communicate, specifically via Ca2+ signaling, within its functional tumour networks may unlock new therapeutics to reduce the rapid infiltration and growth which currently makes treatment ineffective. This work aims to produce patterned networks of GBM cells such that the Ca2+ communication at a network level can be repeatedly and reliably investigated.


The Ethics of Motivational Neuro-Doping in Sport: Praiseworthiness and Prizeworthiness.
Motivational enhancement in sport - a form of 'neuro-doping' - can help athletes attain greater achievements in sport. A key question is whether or not that athlete deserves that achievement. We distinguish three concepts - praiseworthiness (whether the athlete deserves praise), prizeworthiness (whether the athlete deserves the prize), and admiration (pure admiration at the performance) - which are closely related. However, in sport, they can come apart. The most praiseworthy athlete may not be the most prizeworthy, and so on. Using a model of praiseworthiness as costly commitment to a valuable end, and situating prizeworthiness within the boundaries of the sport, we argue that motivational enhancement in some cases can be compatible with desert.


Metabolic enzymes in glial cells of the honeybee brain and their associations with aging, starvation and food response.
The honey bee has been extensively studied as a model for neuronal circuit and memory function and more recently has emerged as an unconventional model in biogerontology. Yet, the detailed knowledge of neuronal processing in the honey bee brain contrasts with the very sparse information available on glial cells. In other systems glial cells are involved in nutritional homeostasis, detoxification, and aging. These glial functions have been linked to metabolic enzymes, such as glutamine synthetase and glycogen phosphorylase. As a step in identifying functional roles and potential differences among honey bee glial types, we examined the spatial distribution of these enzymes and asked if enzyme abundance is associated with aging and other processes essential for survival. Using immunohistochemistry and confocal laser microscopy we demonstrate that glutamine synthetase and glycogen phosphorylase are abundant in glia but appear to co-localize with different glial sub-types. The overall spatial distribution of both enzymes was not homogenous and differed markedly between different neuropiles and also within each neuropil. Using semi-quantitative Western blotting we found that rapid aging, typically observed in shortest-lived worker bees (foragers), was associated with declining enzyme levels. Further, we found enzyme abundance changes after severe starvation stress, and that glutamine synthetase is associated with food response. Together, our data indicate that aging and nutritional physiology in bees are linked to glial specific metabolic enzymes. Enzyme specific localization patterns suggest a functional differentiation among identified glial types.


Effects of calcium on antioxidant activities and water relations associated with heat tolerance in two cool-season grasses.
Calcium (Ca2+) may be involved in plant tolerance to heat stress by regulating antioxidant metabolism or/and water relations. This study was designed to examine whether external Ca2+ treatment would improve heat tolerance in two C(3), cool-season grass species, tall fescue (Festuca arundinacea L.) and Kentucky bluegrass (Poa pratensis L.), and to determine the physiological mechanisms of Ca2+ effects on grass tolerance to heat stress. Grasses were treated with CaCl(2) (10 mM) or H(2)O by foliar application and then exposed to heat stress (35/30 degrees C) in growth chambers. Some of the Ca2+ -untreated plants were maintained at 20/15 degrees C as the temperature control. Heat stress reduced grass quality, relative water content (RWC), and chlorophyll (Chl) content of leaves in both species, but Ca2+ treatment increased all three factors under heat stress. The Ca2+ concentration in cell saps increased with heat stress and with external Ca2+ treatment in both species. Osmotic potential increased with heat stress, but external Ca2+ treatment had no effect. Osmotic adjustment increased during short-term heat stress, but then decreased with a prolonged period of stress; it was not influenced by Ca2+ treatment. The activity of superoxide dismutase (SOD) in both species increased transiently at 12 d of heat stress and then remained at a level similar to that of the control. External Ca2+ treatment had no effect on SOD activity. The activities of catalase (CAT), ascorbate peroxidase (AP), and glutathione reductase (GR) of both species decreased during heat stress. Plants treated with Ca2+ under heat stress had higher CAT, GR and AP activities than untreated plants. Lesser amounts of malondialdehyde (MDA) accumulated in Ca2+ -treated plants than in untreated plants during extended periods of heat stress. The results suggested that exogenous Ca2+ treatment enhanced heat tolerance in both tall fescue and Kentucky bluegrass. This enhancement was related to the maintenance of antioxidant activities and a decrease in membrane lipid peroxidation, but not to the regulation of osmotic potential and osmotic adjustment.


Neural underpinnings of superior action prediction abilities in soccer players.
The ability to form anticipatory representations of ongoing actions is crucial for effective interactions in dynamic environments. In sports, elite athletes exhibit greater ability than novices in predicting other players' actions, mainly based on reading their body kinematics. This superior perceptual ability has been associated with a modulation of visual and motor areas by visual and motor expertise. Here, we investigated the causative role of visual and motor action representations in experts' ability to predict the outcome of soccer actions. We asked expert soccer players (outfield players and goalkeepers) and novices to predict the direction of the ball after perceiving the initial phases of penalty kicks that contained or not incongruent body kinematics. During the task, we applied repetitive transcranial magnetic stimulation (rTMS) over the superior temporal sulcus (STS) and the dorsal premotor cortex (PMd). Results showed that STS-rTMS disrupted performance in both experts and novices, especially in those with greater visual expertise (i.e. goalkeepers). Conversely, PMd-rTMS impaired performance only in expert players (i.e. outfield players and goalkeepers), who exhibit strong motor expertise into facing domain-specific actions in soccer games. These results provide causative evidence of the complimentary functional role of visual and motor action representations in experts' action prediction.


Sensory-Targeted Ankle Rehabilitation Strategies for Chronic Ankle Instability.
Deficient sensory input from damaged ankle ligament receptors is thought to contribute to sensorimotor deficits in those with chronic ankle instability (CAI). Targeting other viable sensory receptors may then enhance sensorimotor control in these patients. The purpose of this randomized controlled trial was to evaluate the effects of 2 wk of sensory-targeted ankle rehabilitation strategies (STARS) on patient- and clinician-oriented outcomes in those with CAI. Eighty patients with self-reported CAI participated. All patients completed patient-oriented questionnaires capturing self-reported function as well as the weight-bearing lunge test and an eyes-closed single-limb balance test. After baseline testing, patients were randomly allocated to four STARS groups: joint mobilization, plantar massage, triceps surae stretching, or control. Each patient in the intervention groups received six 5-min treatments of their respective STARS over 2 wk. All subjects were reassessed on patient- and clinician-oriented measures immediately after the intervention and completed a 1-month follow-up that consisted of patient-oriented measures. Change scores of the three STARS groups were compared with the control using independent t-tests and Hedges' g effect sizes with 95% confidence intervals. The joint mobilization group had the greatest weight-bearing lunge test improvement. Plantar massage had the most meaningful single-limb balance improvement. All STARS groups improved patient-oriented outcomes with joint mobilization having the most meaningful effect immediately after the intervention and plantar massage at the 1-month follow-up. Each STARS treatment offers unique contributions to the patient- and clinician-oriented rehabilitation outcomes of those with CAI. Both joint mobilization and plantar massage appear to demonstrate the greatest potential to improve sensorimotor function in those with CAI.


Fecal Calprotectin as a Marker of the Gut Immune System Activation Is Elevated in Parkinson's Disease.
Alpha-synucleinopathy constituting a characteristic feature of Parkinson's disease (PD) occurs at all levels of the brain-gut axis including the enteric nervous system (ENS). Lesions in the ENS may be connected with gut inflammation, increased intestinal permeability and dysmotility contributing to the pathogenesis of PD and its gastrointestinal manifestations. To evaluate fecal calprotectin and zonulin as biomarkers of gut inflammation and intestinal barrier dysfunction in PD patients. Quantitative evaluation of fecal biomarkers was performed by ELISA tests in 35 PD patients and 20 healthy controls. Additionally, patients filled out a short questionnaire concerning gastrointestinal symptoms. Median fecal calprotectin level (μg/g) was significantly higher in PD patients compared to the controls: 54.5 (29.0-137.9) vs. 9.7 (5.2-23.3), p < 0.0001. Applying age-related reference ranges, the increased fecal calprotectin level was found in 43% of PD patients and in none of the control subjects (p < 0.001). No correlation between fecal calprotectin level and PD duration was observed. No statistically significant difference between the groups regarding zonulin level was found. The most frequent bowel symptoms reported by PD patients included constipation (69% of subjects), feeling of incomplete evacuation (51%), bloating (51%), abdominal pain (20%), and alternating bowel movement pattern (17%). The evaluation of fecal calprotectin level may be a useful tool to detect the signs of gut immune system activation present in a remarkable number of PD patients, also in the early stage of the disease. Calprotectin may constitute a critical link between amyloid formation and neuroinflammatory cascades serving as a prospective diagnostic and therapeutic target.


Surgical management of lumbar spinal stenosis.
Eighty consecutive patients with lumbar spinal stenosis surgically treated during a 5-year period by the author were reviewed. Patients were placed in the following categories: lateral spinal stenosis (10), central-mixed stenosis (29), spinal stenosis after laminectomy and/or fusion (32), and spinal stenosis with degenerative scoliosis (9). Contrast-enhanced computed tomographic (CT) scans were helpful in determining the levels requiring decompression. However, in the multiply operated patient, contrast-enhanced CT scans were misleading in six patients. Patients with lateral spinal stenosis were treated with unilateral laminectomy and partial facetectomy. The 29 patients with central-mixed stenosis underwent decompressive laminectomy and bilateral facetectomies. Six fusions were done. In the nine patients with spinal stenosis and scoliosis, concaveside partial facetectomies and laminectomies were done as well as spinal fusions. The 32 patients with spinal stenosis after previous laminectomy and spinal fusions were the most difficult group to analyze, and their treatment was the least standardized. There were 19 good, eight fair, and five poor results in those who had undergone previous surgery. Fifty-seven of the 80 patients (71%) experienced a good result from their surgical treatment.


Chance-type fractures of the axis.
A retrospective study was performed to identify horizontal fractures of the body of the axis, with special attention to their pattern, prevalence, and clinical outcome. The prevalence of this type of injury and the long-term clinical behavior are examined. Although isolated cases have been reported, horizontal Chance-type fractures of the body of the axis are not common cervical spine injuries. The medical records of 674 consecutive patients with fractures of the cervical spine admitted to the authors' institute from 1970 to 2002 were reviewed. Of them, 2 (0.3%) had a horizontal Chance-type fracture of the body of the axis. Neurologic deficits were not diagnosed at admission. Mechanism of injury, treatment, and long-term follow-up were evaluated. Both patients were treated nonoperatively. At the latest follow-up, 3 and 12 years, respectively, both patients had a satisfactory clinical outcome. Horizontal fractures of the Chance-type of the body of the axis are rare. Conservative treatment proved to be effective.


Minimizing adverse effects on patients of involuntary relocation from long-stay wards to community residences.
This study examined the incidence of, and variables associated with, relocation trauma among 85 patients who moved from long-stay psychiatric wards to community care units as part of a hospital closure in Melbourne, Australia. Some participants moved directly from wards and some moved first to transitional units on the hospital grounds. Preparation for the transition, such as visits to the community care unit before the move, was documented, and severity of symptoms, aggressive behaviors, and preferences for living environments one month before and one month after the move were compared. Relocation trauma was measured one month after the move. After significant factors associated with trauma were determined, additional tests further discriminated these factors. Although 66 percent of participants were happy with community care units one month after the move, 18 percent preferred hospital living. Of 81 participants who had data on relocation trauma, 20, or 25 percent, met relocation trauma criteria. Preparation in a transitional unit reduced the likelihood of trauma, as did making six or more premove visits to the new facility and having a preparation period of more than 16 weeks. When preparation in a transitional unit, number of premove visits, and duration of preparation were considered together, only the number of premove visits and duration of preparation remained significantly associated with relocation trauma. Adverse effects of relocation on long-stay psychiatric patients may be minimized by the preparation of patients over a period of four months or more, with inclusion of six or more visits to the new facility. Moving via a transitional environment is not essential.


Atypical Femur Fracture Risk versus Fragility Fracture Prevention with Bisphosphonates.
Bisphosphonates are effective in reducing hip and osteoporotic fractures. However, concerns about atypical femur fractures have contributed to substantially decreased bisphosphonate use, and the incidence of hip fractures may be increasing. Important uncertainties remain regarding the association between atypical femur fractures and bisphosphonates and other risk factors. We studied women 50 years of age or older who were receiving bisphosphonates and who were enrolled in the Kaiser Permanente Southern California health care system; women were followed from January 1, 2007, to November 30, 2017. The primary outcome was atypical femur fracture. Data on risk factors, including bisphosphonate use, were obtained from electronic health records. Fractures were radiographically adjudicated. Multivariable Cox models were used. The risk-benefit profile was modeled for 1 to 10 years of bisphosphonate use to compare associated atypical fractures with other fractures prevented. Among 196,129 women, 277 atypical femur fractures occurred. After multivariable adjustment, the risk of atypical fracture increased with longer duration of bisphosphonate use: the hazard ratio as compared with less than 3 months increased from 8.86 (95% confidence interval [CI], 2.79 to 28.20) for 3 years to less than 5 years to 43.51 (95% CI, 13.70 to 138.15) for 8 years or more. Other risk factors included race (hazard ratio for Asians vs. Whites, 4.84; 95% CI, 3.57 to 6.56), height, weight, and glucocorticoid use. Bisphosphonate discontinuation was associated with a rapid decrease in the risk of atypical fracture. Decreases in the risk of osteoporotic and hip fractures during 1 to 10 years of bisphosphonate use far outweighed the increased risk of atypical fracture among Whites but less so among Asians. After 3 years, 149 hip fractures were prevented and 2 bisphosphonate-associated atypical fractures occurred in Whites, as compared with 91 and 8, respectively, in Asians. The risk of atypical femur fracture increased with longer duration of bisphosphonate use and rapidly decreased after bisphosphonate discontinuation. Asians had a higher risk than Whites. The absolute risk of atypical femur fracture remained very low as compared with reductions in the risk of hip and other fractures with bisphosphonate treatment. (Funded by Kaiser Permanente and others.).


Macroscopic ionic currents within the human leg.
Recent research on developing and healing tissues suggests that small quasi-DC ionic currents (of magnitude 10-20 microA) may play a controlling role in the initiation and organisation of growing tissues, but the difficulties of measuring such small currents have led to confusing results. Sensitive magnetometry provides a method of demonstrating and, to some extent, locating such currents. A SQUID magnetometer system has been built and used to investigate the magnetic fields around the uninjured human leg. Analysis of the magnetic fields reveals the presence of slowly changing macroscopic current loops (of magnitude up to 12 microA) within the leg. These currents are broadly similar in all subjects, and show day-to-day reproducibility in individuals. They change predictably with time of muscle relaxation (over an hour), and revert to the original form on muscular exertion. These currents are of significance when considering the therapeutic use of injected current for the healing of non-union in bone.


Evaluation and Management of Adolescents With a Stiff Flatfoot.
While flatfeet are normal in children, persistence into adolescence with associated pain or asymmetry warrants additional evaluation. Rigidity of a flatfoot deformity, whether a clinical report or evident on examination, should raise suspicion for pathology. The differential diagnosis includes tarsal coalition, neurogenic planovalgus, and peroneal spasticity. History must include pointed inquiry into birth and neurologic histories to probe for a source of central spasticity. Examination must include standing assessment of hindfoot and midfoot alignment. Hindfoot rigidity may be assessed by the double limb heel rise test and manual examination. Radiographs should include standing ankle (anterior-posterior and mortise) and whole foot (anterior-posterior, external rotation oblique, and lateral) images. Magnetic resonance imaging is more sensitive for identifying coalitions and better characterizes adjacent cartilage, subchondral edema, and tendon pathology, yet CT better characterizes the anatomy of a bony coalition. Conservative treatments are pathology-dependent and play a more prominent role in neurogenic or peroneal spastic flatfoot. Surgical management of coalitions is centered on coalition resection coupled with arthrodesis in the case of a talocalcaneal coalition with a dysplastic subtalar joint; concomitant planovalgus reconstruction is considered on a case-by-case basis.


Characterization of the carbohydrate binding specificity and kinetic parameters of lectins by using surface plasmon resonance.
An accurate, rapid, and sensitive method for characterizing the carbohydrate binding properties of lectins using a BIAcore apparatus and the detection method of surface plasmon resonance is described. As a model study, the sialic acid binding lectins from Sambucus nigra and Maackia amurensis, which are specific for the epitopes Neu5Ac(alpha2-6)Gal and Neu5Ac(alpha2-3)Gal, respectively, were chosen as suitable candidates. Two systems, one for the analysis of oligosaccharides and the other for glycoproteins, were developed after a rigorous analysis and evaluation of such parameters as binding conditions, buffers, and regeneration conditions. The systems take into account nonspecific binding, using the respective denatured lectin as negative blank, and avoid loss of activity: regeneration of the surface using either 10 mM NaOAc (pH 4.3) buffer (oligosaccharide system) or 20 mM HCl (glycoprotein system). The specificity of the lectins is well illustrated, while the kinetics parameters are shown to be sensitive to subtle changes in the recognized epitopes, and to be affected by steric hindrance. Surface plasmon resonance is a suitable technique for the analysis and characterization of lectins.


Geldanamycin, radicicol, and chimeric inhibitors of the Hsp90 N-terminal ATP binding site.
Natural products have continued to drive the development of new chemotherapeutics and elucidation of new biological targets for the treatment of disease. Since Whitesell and Neckers' original discovery that geldanamycin does not directly inhibit v-Src, but instead manifests its biological activity through inhibition of the Hsp90 molecular chaperone, additional natural products and natural product derivatives have been identified and developed to inhibit the Hsp90 protein folding machinery. 17-AAG, a geldanamycin analogue, is currently in clinical trials for the treatment of several types of cancer. Recent work has produced improved radicicol analogues that show promising Hsp90 inhibitory activity in vitro. In addition, chimeric molecules of these two natural products are active in vitro and represent a novel class of Hsp90 inhibitors for cancer treatment. In addition to their chemotherapeutic uses, natural product inhibitors and their derivatives have been utilized to probe the biological mechanisms by which Hsp90 inhibition regulates tumor cell growth. As a consequence of these studies, the molecular chaperones have emerged as an exciting new class of therapeutic targets. This review will highlight the utility of the natural products, geldanamycin and radicicol, as well as improved analogues and the activities exhibited by these compounds against various cancer cell lines.


[Local administration of metronidazole before hysterectomy--effect on infectious morbidity. A prospective study].
A prospective clinical trial was designed to examine the influence of a local application of metronidazole on postoperative infectious morbidity in 134 women undergoing vaginal and in 108 women undergoing abdominal hysterectomy. 116 patients received on the evening before hysterectomy a vaginal suppository of metronidazole (500 mg). The control group (126 patients) had no metronidazole prophylaxis. All patients, who underwent vaginal hysterectomy, were given additionally a single-shot prophylaxis with 1500 mg cefuroxim. With regard to febrile morbidity, urinary tract infection and abdominal wound or pelvic infection, there was a significant decrease in infectious morbidity. In women undergoing vaginal hysterectomy the infectious morbidity was reduced from 35.7% to 20.3% (p less than 0.05) in spite of systemic prophylaxis. In cases with abdominal hysterectomy, we saw a decrease from 64.3% to 26.9% (p less than 0.001). There was no effect on the incidence of urinary tract infections. Local prophylaxis with metronidazole in abdominal hysterectomy and additional to a systemic prophylaxis in vaginal hysterectomy reduces the infectious morbidity in our patients.


Laser Capture Microdissection of Pancreatic Acinar Cells to Identify Proteomic Alterations in a Murine Model of Caerulein-Induced Pancreatitis.
Chronic pancreatitis (CP) is characterized by inflammation and fibrosis of the pancreas, leading to pain, parenchymal damage, and loss of exocrine and endocrine function. There are currently no curative therapies; diagnosis remains difficult and aspects of pathogenesis remain unclear. Thus, there is a need to identify novel biomarkers to improve diagnosis and understand pathophysiology. We hypothesize that pancreatic acinar regions contain proteomic signatures relevant to disease processes, including secreted proteins that could be detected in biofluids. Acini from pancreata of mice injected with or without caerulein were collected using laser capture microdissection followed by mass spectrometry analysis. This protocol enabled high-throughput analysis that captured altered protein expression throughout the stages of CP. Over 2,900 proteins were identified, whereas 331 were significantly changed ≥2-fold by mass spectrometry spectral count analysis. Consistent with pathogenesis, we observed increases in proteins related to fibrosis (e.g., collagen, P<0.001), several proteases (e.g., trypsin 1, P<0.001), and altered expression of proteins associated with diminished pancreas function (e.g., lipase, amylase, P<0.05). In comparison with proteomic data from a public data set of CP patients, a significant correlation was observed between proteomic changes in tissue from both the caerulein model and CP patients (r=0.725, P<0.001). This study illustrates the ability to characterize proteome changes of acinar cells isolated from pancreata of caerulein-treated mice and demonstrates a relationship between signatures from murine and human CP.


Surface parameterization in volumetric images for curvature-based feature classification.
Curvature-based surface features are well suited for use in multimodal medical image registration. The accuracy of such feature-based registration techniques is dependent upon the reliability of the feature computation. The computation of curvature features requires second derivative information that is best obtained from a parametric surface representation. We present a method of explicitly parameterizing surfaces from volumetric data. Surfaces are extracted, without a global thresholding, using active contour models. A monge/spl acute/ basis for each surface patch is estimated and used to transform the patch into local, or parametric, coordinates. Surface patches are fit to a bicubic polynomial in local coordinates using least squares solved by singular value decomposition. We tested our method by reconstructing surfaces from the surface model and analytically computing Gaussian and mean curvatures. The model was tested on analytical and medical data.


The Influence of Socioeconomic Factors on Access to Biologics in Psoriasis.
Since the introduction of biologics for psoriasis, uptake has been uneven and limited. Few studies have investigated the influence of socioeconomic factors on access to biologics. To investigate how socioeconomic factors influenced access to biologics. Biologic-naïve patients in the Swedish National Register for Systemic Treatment of Psoriasis (PsoReg) for the years 2006-2014 were included. For patients who remained on nonbiologic treatments during their entire registration (n = 1851), the most recent registration was analyzed. For patients who began treatment with biologics during registration in PsoReg (n = 665), the last observation before initiation of biologics was analyzed. A logistic regression model was used to investigate whether education and income influenced the probability of a switch to biologics, whilst adjusting for demographic and individual factors such as age, sex, disease severity, and clinical characteristics. The odds ratio of access to biologics was 1.8 (CI = 1.3-2.6) in the group with a high level of disposable income, compared with the middle-income group. No differences were found concerning educational levels. The odds ratios of access to biologics decreased with age. Patients with psoriatic arthritis had odds ratios of access to biologics which were more than 50 percent higher, controlling for other variables. High disease severity, in terms of physician- and patient-reported severity, increased the odds ratios of access to biologics. The higher-income group had better access to biologics than the middle-income group when adjusting for disease severity and lifestyle factors. This may not only be an equity problem, as a better allocation of society's resources might have resulted in a higher overall effectiveness of biologics.


Performance Evaluation of Montelukast Pediatric Formulations: Part I-Age-Related In Vitro Conditions.
This study aimed to explore the potential of biopharmaceutics in vitro tools to predict drug product performance in the pediatric population. Biorelevant dissolution set-ups were used to predict how age and medicine administration practices affect the in vitro dissolution of oral formulations of a poorly water-soluble compound, montelukast. Biorelevant age-appropriate dissolution studies of Singulair® (granules and chewable tablets) were conducted with the µDISS profiler™, USP 4 apparatus, USP 2 apparatus, and mini-paddle apparatus. Biorelevant simulating fluids representative of adult and pediatric conditions were used in the dissolution studies. The biorelevant dissolution conditions were appropriately selected (i.e. volumes, transit times, etc.) to mimic the gastrointestinal conditions of each of the subpopulations tested. Partial least squares regression (PLS-R) was performed to understand the impact of in vitro variables on the dissolution of montelukast. Montelukast dissolution was significantly affected by the in vitro hydrodynamics used to perform the dissolution tests (µDISS profiler™: positive effect); choice of simulation of gastric (negative effect) and/or intestinal conditions (positive effect) of the gastrointestinal tract; and simulation of prandial state (fasted state: negative effect, fed state: positive effect). Age-related biorelevant dissolution of Singulair® granules predicted the in vivo effect of the co-administration of the formulation with applesauce and formula in infants. This study demonstrates that age-appropriate biorelevant dissolution testing can be a valuable tool for the assessment of drug performance in the pediatric population.


Angiotensin-(1-9) prevents cardiomyocyte hypertrophy by controlling mitochondrial dynamics via miR-129-3p/PKIA pathway.
Angiotensin-(1-9) is a peptide from the noncanonical renin-angiotensin system with anti-hypertrophic effects in cardiomyocytes via an unknown mechanism. In the present study we aimed to elucidate it, basing us initially on previous work from our group and colleagues who proved a relationship between disturbances in mitochondrial morphology and calcium handling, associated with the setting of cardiac hypertrophy. Our first finding was that angiotensin-(1-9) can induce mitochondrial fusion through DRP1 phosphorylation. Secondly, angiotensin-(1-9) blocked mitochondrial fission and intracellular calcium dysregulation in a model of norepinephrine-induced cardiomyocyte hypertrophy, preventing the activation of the calcineurin/NFAT signaling pathway. To further investigate angiotensin-(1-9) anti-hypertrophic mechanism, we performed RNA-seq studies, identifying the upregulation of miR-129 under angiotensin-(1-9) treatment. miR-129 decreased the transcript levels of the protein kinase A inhibitor (PKIA), resulting in the activation of the protein kinase A (PKA) signaling pathway. Finally, we showed that PKA activity is necessary for the effects of angiotensin-(1-9) over mitochondrial dynamics, calcium handling and its anti-hypertrophic effects.


Factors associated with preterm delivery and low birth weight: a study from rural Maharashtra, India.
Background: Although preterm delivery and low birth weight (LBW) have been studied in India, findings may not be generalisable to rural areas such as the Marathwada region of Maharashtra state. There is limited information available on maternal and child health indicators from this region. We aimed to present some local estimates of preterm delivery and LBW in the Osmanabad district of Marathwada and assess available maternal risk factors. Methods: The study used routinely collected data on all in-hospital births in the maternity department of Halo Medical Foundation's hospital from 1 st January 2008 to 31 st December 2014. Multivariable logistic regression analysis provided odds ratios (OR) with 95% confidence intervals (CI) for preterm delivery and LBW according to each maternal risk factor. Results: We analysed 655 live births, of which 6.1% were preterm deliveries. Of the full term births (N=615), 13.8% were LBW (<2.5 kilograms at birth). The odds of preterm delivery were three times higher (OR=3.23, 95% CI 1.36 to 7.65) and the odds of LBW were double (OR=2.03, 95% CI 1.14 to 3.60) among women <22 years of age compared with older women. The odds of both preterm delivery and LBW were reduced in multigravida compared with primigravida women regardless of age. Anaemia (Hb<11g/dl), which was prevalent in 91% of women tested, was not significantly related to these birth outcomes. Conclusions: The odds of preterm delivery and LBW were much higher in mothers under 22 years of age in this rural Indian population. Future studies should explore other related risk factors and the reasons for poor birth outcomes in younger mothers in this population, to inform the design of appropriate public health policies that address this issue.


High-Order Visual Processing, Visual Symptoms, and Visual Hallucinations: A Possible Symptomatic Progression of Parkinson's Disease.
Objective: To determine whether Parkinson disease (PD) patients with (VH) have different clinical characteristics and gray-matter volume than those with visual misperceptions (VM) or other visual symptoms (OvS). Background: The spectrum of visual complaints in PD is broad and complex. Methods: We conducted a retrospective chart review of 525 PD patients to identify the frequency of visual symptoms and the association with clinical and radiological features. Brain volumetric MRI data was analyzed using multivariate logistic regression to differentiate cases with and without visual symptoms. Results: Among 525 PD cases, visual complaints were documented in 177 (33.7%). Among these, 83 (46.9%) had VH, 31 (17.5%) had VM, and 63 (35.6%) had OvS (diplopia, blurry vision, photophobia, dry eyes, and eye pain or soreness). When compared to OvS, patients with VH had significantly higher age, duration of disease, rate of REM sleep behavior disorder, and cognitive impairment. Visual hallucinations patients had decreased age-adjusted volumetric averages in 28/30 gray-matter regions when compared to PD without visual symptoms and 30/30 gray-matter regions when compared to VM patients. Conclusions: Visual symptoms in PD may represent a spectrum from OvS to VM to VH, with progression of the latter associated with older age, duration of disease, presence of REM sleep behavior disorder, cognitive impairment, and decreased gray-matter volume.


Temperature stress mediates decanalization and dominance of gene expression in Drosophila melanogaster.
The regulatory architecture of gene expression remains an area of active research. Here, we studied how the interplay of genetic and environmental variation affects gene expression by exposing Drosophila melanogaster strains to four different developmental temperatures. At 18°C we observed almost complete canalization with only very few allelic effects on gene expression. In contrast, at the two temperature extremes, 13°C and 29°C a large number of allelic differences in gene expression were detected due to both cis- and trans-regulatory effects. Allelic differences in gene expression were mainly dominant, but for up to 62% of the genes the dominance swapped between 13 and 29°C. Our results are consistent with stabilizing selection causing buffering of allelic expression variation in non-stressful environments. We propose that decanalization of gene expression in stressful environments is not only central to adaptation, but may also contribute to genetic disorders in human populations.


Changing substrate specificity and iteration of amino acid chain elongation in glucosinolate biosynthesis through targeted mutagenesis of Arabidopsis methylthioalkylmalate synthase 1.
Methylthioalkylmalate synthases catalyse the committing step of amino acid chain elongation in glucosinolate biosynthesis. As such, this group of enzymes plays an important role in determining the glucosinolate composition of Brassicaceae species, including Arabidopsis thaliana Based on protein structure modelling of MAM1 from A. thaliana and analysis of 57 MAM sequences from Brassicaceae species, we identified four polymorphic residues likely to interact with the 2-oxo acid substrate. Through site-directed mutagenesis, the natural variation in these residues and the effect on product composition were investigated. Fifteen MAM1 variants as well as the native MAM1 and MAM3 from A. thaliana were characterised by heterologous expression of the glucosinolate chain elongation pathway in Escherichia coli Detected products derived from leucine, methionine or phenylalanine were elongated with up to six methylene groups. Product profile and accumulation were changed in 14 of the variants, demonstrating the relevance of the identified residues. The majority of the single amino acid substitutions decreased the length of methionine-derived products, while approximately half of the substitutions increased the phenylalanine-derived products. Combining two substitutions enabled the MAM1 variant to increase the number of elongation rounds of methionine from three to four. Notably, characterisation of the native MAMs indicated that MAM1 and not MAM3 is responsible for homophenylalanine production. This hypothesis was confirmed by glucosinolate analysis in mam1 and mam3 mutants of A. thaliana.


Cadmium inhibits vacuolar H(+)ATPase-mediated acidification in the rat epididymis.
In rats, an acidic luminal pH maintains sperm quiescence during storage in the epididymis. We recently showed that vacuolar H(+)ATPase-rich cells in the epididymis and vas deferens are involved in the acidification of these segments. Treatment of rats with cadmium (Cd) leads to alkalinization of this fluid by an unknown mechanism. Because Cd may affect H(+)ATPase function, we examined 1) the in vivo effect of Cd poisoning on H(+)ATPase-rich cell morphology and on the abundance and distribution of the 31-kDa H(+)ATPase subunit in cells along the rat epididymis, and 2) the in vitro effect of Cd on H(+)ATPase activity and function in the isolated vas deferens. Immunofluorescence and immunoblotting data from rats treated with Cd for 14-15 days (2 mg Cd/kg body mass/day) showed that 1) H(+)ATPase-positive cells regressed to a prepubertal phenotype, and 2) H(+)ATPase was lost from the apical pole of the cell and was redistributed into an intracellular compartment. In experiments in vitro, Cd inhibited bafilomycin-sensitive ATPase activity in isolated total cell membranes and, as measured using a proton-selective extracellular microelectrode, inhibited proton secretion in isolated vas deferens. We conclude that alkalinization of the tubule fluid in the epididymis and vas deferens of Cd-treated rats may result from the loss of functional H(+)ATPase enzyme in the cell apical domain as well as from a direct inhibition of H(+)ATPase function by Cd.


Studying zebrafish nervous system structure and function in health and disease with electron microscopy.
Zebrafish (Danio rerio) is a well-established model for studying the nervous system. Findings in zebrafish often inform studies on human diseases of the nervous system and provide crucial insight into disease mechanisms. The functions of the nervous system often rely on communication between neurons. Signal transduction is achieved via release of signaling molecules in the form of neuropeptides or neurotransmitters at synapses. Snapshots of membrane dynamics of these processes are imaged by electron microscopy. Electron microscopy can reveal ultrastructure and thus synaptic processes. This is crucial both for mapping synaptic connections and for investigating synaptic functions. In addition, via volumetric electron microscopy, the overall architecture of the nervous system becomes accessible, where structure can inform function. Electron microscopy is thus of particular value for studying the nervous system. However, today a plethora of electron microscopy techniques and protocols exist. Which technique is most suitable highly depends on the research question and scope as well as on the type of tissue that is examined. This review gives an overview of the electron microcopy techniques used on the zebrafish nervous system. It aims to give researchers a guide on which techniques are suitable for their specific questions and capabilities as well as an overview of the capabilities of electron microscopy in neurobiological research in the zebrafish model.


[Results of the sequential use of Mexidol and Mexidol Forte 250 in patients with chronic cerebral ischemia].
To study the efficacy and safety of mexidol's intravenous injections (500 mg once a day) for 14 days, followed by oral administration of mexidol FORTE 250 at a dose of 250 mg (1 tablet) 3 times a day for 60 days, in treatment of chronic cerebral ischemia (CCI) in patients with hypertension and atherosclerosis of the brachiocephalic arteries. The observation program included 60 patients with an established diagnosis of CCI confirmed by neuroimaging methods. Patients of the main group (n=26) received mexidol along with basic therapy, patients of the comparison group (n=26) received only basic therapy. The results of the experience show the high efficacy and safety of sequential therapy (parenteral therapy followed by tablets of mexidol FORTE 250). The treatment improves emotional and cognitive status, decreases static-motor disorders and severity of subjective neurological symptoms. High adherence of patients to the therapy is shown.


Intracellular trafficking as a determinant of AS-DACA cytotoxicity in rhabdomyosarcoma cells.
Rhabdomyosarcoma (RMS) is a malignant soft tissue sarcoma derived from skeletal muscle precursor cells, which accounts for 5-8% of all childhood malignancies. Disseminated RMS represents a major clinical obstacle, and the need for better treatment strategies for the clinically aggressive alveolar RMS subtype is particularly apparent. Previously, we have shown that the acridine-4-carboxamide derivative AS-DACA, a known topoisomerase II poison, is potently cytotoxic in the alveolar RMS cell line RH30, but is 190-fold less active in the embryonal RMS cell line RD. Here, we investigate the basis for this selectivity, and demonstrate in these RMS lines, and in an AS-DACA- resistant subclone of RH30, that AS-DACA-induced cytotoxicity correlates with the induction of DNA double strand breaks. We show that inhibition of the multidrug-resistance associated protein (MRP1) has no effect on AS-DACA sensitivity. By exploiting the pH-dependent fluorescence properties of AS-DACA, we have characterized its intracellular distribution, and show that it concentrates in the cell nucleus, as well as in acidic vesicles of the membrane trafficking system. We show that fluorescence microscopy can be used to determine the localization of AS-DACA to the nuclear and cytoplasmic compartments of RMS cells grown as spheroids, penetrance being much greater in RH30 than RD spheroids, and that the vesicular signal leads the way into the spheroid mass. EEA1 and Rab5 proteins, molecular markers expressed on early-endosomal vesicles, are reduced by >50% in the sensitive cell lines. Taking the evidence as a whole, suggests that endosomal vesicle trafficking influences the toxicity of AS-DACA in RMS cells.


Long-term adaptation of epistatic genetic networks.
Gene networks are likely to govern most traits in nature. Mutations at these genes often show functional epistatic interactions that lead to complex genetic architectures and variable fitness effects in different genetic backgrounds. Understanding how epistatic genetic systems evolve in nature remains one of the great challenges in evolutionary biology. Here we combine an analytical framework with individual-based simulations to generate novel predictions about long-term adaptation of epistatic networks. We find that relative to traits governed by independently evolving genes, adaptation with epistatic gene networks is often characterized by longer waiting times to selective sweeps, lower standing genetic variation, and larger fitness effects of adaptive mutations. This may cause epistatic networks to either adapt more slowly or more quickly relative to a nonepistatic system. Interestingly, epistatic networks may adapt faster even when epistatic effects of mutations are on average deleterious. Further, we study the evolution of epistatic properties of adaptive mutations in gene networks. Our results show that adaptive mutations with small fitness effects typically evolve positive synergistic interactions, whereas adaptive mutations with large fitness effects evolve positive synergistic and negative antagonistic interactions at approximately equal frequencies. These results provide testable predictions for adaptation of traits governed by epistatic networks and the evolution of epistasis within networks.


Cardiovascular physiology and sleep.
Sleep is a natural periodic suspension of consciousness during which processes of rest and restoration occur. The cognitive, reparative and regenerative accompaniments of sleep appear to be essential for maintenance of health and homeostasis. This brief overview will examine the cardiovascular responses to normal and disordered sleep, and their physiologic and pathologic implications. In the past, sleep was believed to be a passive state. The tableau of sleep as it unfolds is anything but a passive process. The brain's activity is as complex as wakefulness, never "resting" during sleep. Following the demise of the 'passive theory of sleep' (the reticular activating system is fatigued during the waking day and hence becomes inactive), there arose the 'active theory of sleep' (sleep is due to an active general inhibition of the brain) (1). Hess demonstrated the active nature of sleep in cats, inducing "physiological sleep" with electrical stimulation of the diencephalon (2). Classical experiments of transection of the cat brainstem (3) at midpontine level inhibited sleep completely, implying that centers below this level were involved in the induction of sleep (1, 4). For the first time, measurement of sleep depth without awakening the sleeper using the electroencephalogram (EEG) was demonstrated in animals by Caton and in humans, by Berger (1). This was soon followed by discovery of the rapid eye movement sleep periods (REM) by Aserinski and Kleitman (5), demonstration of periodical sleep cycles and their association with REM sleep (6, 7). Multiple studies and steady discoveries (4) made polysomnography, with its ability to perform simultaneous whole night recordings of EEG, electromyogram (EMG), and electrooculogram (EOC), a major diagnostic tool in study of sleep disorders. This facility has been of further critical importance in allowing evaluation of the interaction between sleep and changes in hemodynamics and autonomic cardiovascular control. Consequently the effects of sleep could be objectively differentiated from the effects of rest and recumbency. Furthermore, the specific effects of sleep onset and termination, and the effects of different sleep stages, could be assessed. Technological advances, with consequently enhanced and relatively non-invasive approaches to cardiovascular regulation, have greatly broadened our understanding of the effects of sleep stage on cardiovascular function. Continuous monitoring of simultaneous measures of polysomnographic and cardiovascular variables enables characterization of the effects of dynamic changes and rapid transitions in sleep stage, such as arousals. The capacity for measuring acute and immediate changes in autonomic, EEG and hemodynamic responses to sleep and arousal on a continuous basis has played an important role in enabling us to understand the interplay between changes in EEG and changes in the more peripheral measurements of neural and circulatory variables, such as sympathetic nerve traffic, heart rate (HR) and blood pressure (BP). Measurements of heart rate variability (HRV) (8-10), baroreflex sensitivity (BRS) (11-16), and intraneural measurement of sympathetic nerve traffic to muscle (MSNA) (17-22) and skin (SSNA) (23-24) have further advanced our understanding of mechanisms linking sleep and cardiovascular physiology.


Multi-modal delivery of therapeutics using biomaterial scaffolds.
Functionalisation of biomaterials with therapeutic moieties (proteins, drugs, genes) is a pre-requisite to tissue regeneration and restoration of function following injury or disease. However, up until now, single-factor delivery has not proven to be clinically efficacious, most likely due to the complex nature of pathological states. In this regard, strategies that respect the complex nature of disease can prove successful, paving the way for the delivery of several factors to modulate several stages of the pathology over time. Biomaterials offer opportunities to deliver multiple therapeutics in a temporal manner (multi-modal release) using a number of strategies. The importance of these strategies will be described, as well as the methodologies used to achieve multi-modal release. Furthermore, strategies to engineer more programmed and responsive biomaterials as multi-modal delivery systems will be explored.


Comparison of house dust mite sensitization profiles in allergic adults from Canada, Europe, South Africa and USA.
Sensitization to house dust mite (HDM) is a leading cause of allergic rhinitis and asthma. Despite more than 30 HDM-derived allergens having been identified to date, specific therapeutic approaches do not yet take into account the local sensitization profiles of patients. This study aimed to identify patterns of HDM sensitization in HDM-allergic adults living in distinct geographic areas, to inform the development of targeted diagnostic and therapeutic tools. Serum samples from 685 HDM-allergic subjects from Canada, Europe, South Africa, and the USA were tested for levels of IgE specific for 17 micro-arrayed HDM allergens by ImmunoCAP Immuno Solid-phase Allergen Chip (ISAC) technology. The results confirmed significant geographical variability in sensitization patterns and levels of IgE. In all areas, the major sensitizers were the group 1 and group 2 allergens and Der p 23. Der p 23 was a frequent sensitizer: 64% of the subjects had IgE specific for Der p 23, and 2.3% were monosensitized to it. In South Africa, Der p 23 was the dominant HDM allergen (86% prevalence) and Der p 7 achieved major allergen status (56%). IgE sensitization to HDM was influenced by asthmatic status, levels of allergen exposure, age, race-ethnicity and smoking status, but not by BMI. Sensitization profiles to HDM allergens differ considerably among distinct geographic areas, with Der p 7 and Der p 23 being major sensitizers in South Africa. Such heterogeneity should be taken into account in the diagnosis and treatment of HDM-allergic patients.


H4IIE rat hepatoma cell bioassay-derived 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents in colonial fish-eating waterbird eggs from the Great Lakes.
Fish-eating waterbirds from the Great Lakes of North America have shown symptoms of poisoning similar to those observed in laboratory exposures of various avian species to planar halogenated hydrocarbons (PHHs). PHHs, include among others, polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins (PCDDs), and polychlorinated dibenzofurans (PCDFs), and have been implicated in some of the reproductive problems of Great Lakes water-birds. The objectives of this study were to assess the overall potencies of PCB-containing extracts from colonial water-bird eggs taken from the Great Lakes and to compare the potencies with the location and spatial distribution of the colonies. The potencies of the extracts were assessed by their ability to induce cytochrome P450IA1-associated ethoxyresorufin O-deethylase (EROD) activity in H4IIE rat hepatoma cells as compared to the standard, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). The H4IIE bioassay-derived TCDD-equivalents (TCCD-EQs) in the waterbird eggs concur with residue analyses and biological data from other studies. The greatest concentrations of TCDD-EQs were found in waterbird eggs from historically polluted, industrialized or urbanized areas in which the reproductive impairment of colonial waterbirds was most severe. However, significant concentrations of TCDD-EQs were detected at all sites tested; with a range of 49 to 415 pg TCDD-EQ/g egg, uncorrected for extraction efficiencies. The H4IIE bioassay proved to be a useful biomonitoring tool to assess the overall potency of complex PHH mixtures in environmental samples.


[The prostaglandin and cyclic nucleotide levels in the organs of mice with experimental anthrax intoxication].
In this work the influence of Bacillus anthracis toxin, introduced intraperitoneally in a dose of LD100, on the content of prostaglandins E and F2 alpha, 6-ketoprostaglandin F1 alpha, thromboxane, cAMP and cGMP in the lungs, heart, liver and spleen of BALB/c mice in the time course of experimental intoxication has been studied. The concentration and proportion of prostaglandins and cyclic nucleotides have been shown to undergo-sharp changes in all organs under study in the process of intoxication. The level and proportion of prostaglandins in the lungs ensures the development of vaso- and bronchodilatation processes even at early stages of the action of the toxin. B. anthracis toxin sharply increases the content of cGMP in the organs under study and cAMP in the liver. The activating effect on the adenylate cyclase system of tissue cells is not linked with the action of the edematous factor of the toxin. The role of cyclic nucleotides and prostaglandins in the development of pulmonary edema in intoxication with B. anthracis toxin is discussed.


Can right-sided atrioventricular sequential pacing provide benefit for selected patients with severe congestive heart failure?
This study reports on the methods and results of applying right-sided atrioventricular (AV) pacing in 26 patients with advanced cardiomyopathy. Ten of these patients had ischemic cardiomyopathy. Of the 16 patients with nonischemic cardiomyopathy, 10 were idiopathic and 6 were due to secondary causes. The patients had a mean age of 56 +/- 12 years and a left ventricular ejection fraction of 26 +/- 11%. Two transvenous stimulation electrodes were temporarily placed in the high right atrium and right ventricle, respectively. A Swan Ganz catheter was positioned into the pulmonary artery to determine cardiac output by the thermodilution method and to measure the pressure in the pulmonary artery and right atrium. In addition, aortic pressure was measured through a catheter sheath via the right femoral artery. Systemic and pulmonary vascular resistance were calculated. Stimulation was performed in VVI and DDD pacing modes using different AV intervals (40, 80, 125, 150, 175, 200, and 250 msec). No increase of cardiac output was observed for the overall study cohort (p = 0.51). At VVI pacing, stroke volume significantly decreased from 66 +/- 20 mL to 53 +/- 13 mL (p < 0.01). We distinguished between responders who developed an increase of cardiac output of > 1 L/min (n = 12, 46%) and nonresponders (n = 14, 54%). Etiology of either ischemic or nonischemic cardiomyopathy for responders, as well as conduction disturbances (first-degree AV block, LBBB, RBBB), were equally distributed among both groups. Using an AV delay of 150 and 175 msec, responders to DDD pacing derived a significant increase in cardiac output. An AV delay of 150 msec produced both a significant increase of stroke volume and decrease of systemic vascular resistance. In 46% of patients with dilated cardiomyopathy of either ischemic or nonischemic origin, right-sided AV-sequential pacing brought about an improvement of left ventricular function in terms of enhanced cardiac output. We suggest individual testing in all patients with severe left ventricular dysfunction to find responders.


Combination of voriconazole and anidulafungin for treatment of triazole-resistant aspergillus fumigatus in an in vitro model of invasive pulmonary aspergillosis.
Voriconazole is a first-line agent for the treatment of invasive pulmonary aspergillosis. Isolates with elevated voriconazole MICs are increasingly being seen, and the optimal treatment regimen is not defined. We investigated whether the combination of voriconazole with anidulafungin may be beneficial for the treatment of A. fumigatus strains with elevated voriconazole MICs. We used an in vitro model of the human alveolus to define the exposure-response relationships for a wild-type strain (voriconazole MIC, 0.5 mg/liter) and strains with defined molecular mechanisms of triazole resistance (MICs, 4 to 16 mg/liter). All strains had anidulafungin minimum effective concentrations (MECs) of 0.0078 mg/liter. Exposure-response relationships were estimated using galactomannan as a biomarker. Concentrations of voriconazole and anidulafungin were measured using high-performance liquid chromatography (HPLC). The interaction of voriconazole and anidulafungin was described using the Greco model. Fungal growth was progressively inhibited with higher drug exposures of voriconazole. Strains with elevated voriconazole MICs required proportionally greater voriconazole exposures to achieve a comparable antifungal effect. Galactomannan concentrations were only marginally reduced by anidulafungin monotherapy. An additive effect between voriconazole and anidulafungin was apparent. In conclusion, the addition of anidulafungin does not markedly alter the exposure-response relationship of voriconazole. A rise in serum galactomannan during combination therapy with voriconazole and anidulafungin should be interpreted as treatment failure and not attributed to a paradoxical reaction related to echinocandin treatment.


Engage In or Refrain From? A Qualitative Exploration of Premarital Sexual Relations Among Female College Students in Tehran.
This study explored the factors that educated young Iranian women consider when they decide whether to have premarital sex. Using a purposive sampling method, 30 in-depth interviews were conducted with female college students in both government and private universities in Tehran in 2005-2006. The respondents included unmarried women who had experienced sexual relations, unmarried women who had abstained, and married women. Young women's considerations for premarital sex included (1) marriage meaning and motivations; (2) compliance with family values and expectations; (3) perceived gender and social norms of premarital sex; (4) importance of religiosity; and (5) sexual knowledge and self-efficacy. Marriage meaning and motivation seem to be the central pillar in the complex decision to engage in or refrain from premarital sex among female college students. These considerations have theoretical implications for understanding premarital relationships and sex in a conservative setting. Finally, some shifts are occurring in the meaning of marriage and sexual mores among educated young women in Iran; these changes are discussed.


Clinical and endoscopic predictors of cytological dysplasia or cancer in a prospective multicentre study of large sessile serrated adenomas/polyps.
The serrated neoplasia pathway accounts for up to 30% of all sporadic colorectal cancers (CRCs). Sessile serrated adenomas/polyps (SSA/Ps) with cytological dysplasia (SSA/P-D) are a high-risk serrated CRC precursor with little existing data. We aimed to describe the clinical and endoscopic predictors of SSA/P-D and high grade dysplasia (HGD) or cancer. Prospective multicentre data of SSA/Ps ≥20 mm referred for treatment by endoscopic mucosal resection (September 2008-July 2013) were analysed. Imaging and lesion assessment was standardised. Histological findings were correlated with clinical and endoscopic findings. 268 SSA/Ps were found in 207/1546 patients (13.4%). SSA/P-D comprised 32.4% of SSA/Ps ≥20 mm. Cancer occurred in 3.9%. On multivariable analysis, SSA/P-D was associated with increasing age (OR=1.69 per decade; 95% CI (1.19 to 2.40), p0.004) and increasing lesion size (OR=1.90 per 10 mm; 95% CI (1.30 to 2.78), p0.001), an 'adenomatous' pit pattern (Kudo III, IV or V) (OR=3.98; 95% CI (1.94 to 8.15), p<0.001) and any 0-Is component within a SSA/P (OR=3.10; 95% CI (1.19 to 8.12) p0.021). Conventional type dysplasia was more likely to exhibit an adenomatous pit pattern than serrated dysplasia. HGD or cancer was present in 7.2% and on multivariable analysis, was associated with increasing age (OR=2.0 per decade; 95% CI 1.13 to 3.56) p0.017) and any Paris 0-Is component (OR=10.2; 95% CI 3.18 to 32.4, p<0.001). Simple assessment tools allow endoscopists to predict SSA/P-D or HGD/cancer in SSA/Ps ≥20 mm. Correct prediction is limited by failure to recognise SSA/P-D which may mimic conventional adenoma. Understanding the concept of SSA/P-D and the pitfalls of SSA/P assessment may improve detection, recognition and resection and potentially reduce interval cancer. NCT01368289.


Intravascular donor monocytes play a central role in lung transplant ischaemia-reperfusion injury.
Primary graft dysfunction in lung transplant recipients derives from the initial, largely leukocyte-dependent, ischaemia-reperfusion injury. Intravascular lung-marginated monocytes have been shown to play key roles in experimental acute lung injury, but their contribution to lung ischaemia-reperfusion injury post transplantation is unknown. To define the role of donor intravascular monocytes in lung transplant-related acute lung injury and primary graft dysfunction. Isolated perfused C57BL/6 murine lungs were subjected to warm ischaemia (2 hours) and reperfusion (2 hours) under normoxic conditions. Monocyte retention, activation phenotype and the effects of their depletion by intravenous clodronate-liposome treatment on lung inflammation and injury were determined. In human donor lung transplant samples, the presence and activation phenotype of monocytic cells (low side scatter, 27E10+, CD14+, HLA-DR+, CCR2+) were evaluated by flow cytometry and compared with post-implantation lung function. In mouse lungs following ischaemia-reperfusion, substantial numbers of lung-marginated monocytes remained within the pulmonary microvasculature, with reduced L-selectin and increased CD86 expression indicating their activation. Monocyte depletion resulted in reductions in lung wet:dry ratios, bronchoalveolar lavage fluid protein, and perfusate levels of RAGE, MIP-2 and KC, while monocyte repletion resulted in a partial restoration of the injury. In human lungs, correlations were observed between pre-implantation donor monocyte numbers/their CD86 and TREM-1 expression and post-implantation lung dysfunction at 48 and 72 hours. These results indicate that lung-marginated intravascular monocytes are retained as a 'passenger' leukocyte population during lung transplantation, and play a key role in the development of transplant-associated ischaemia-reperfusion injury.


Introducing 4s-2p Orbital Hybridization to Stabilize Spinel Oxide Cathodes for Lithium-Ion Batteries.
Oxides composed of an oxygen framework and interstitial cations are promising cathode materials for lithium-ion batteries. However, the instability of the oxygen framework under harsh operating conditions results in fast battery capacity decay, due to the weak orbital interactions between cations and oxygen (mainly 3d-2p interaction). Here, a robust and endurable oxygen framework is created by introducing strong 4s-2p orbital hybridization into the structure using LiNi0.5 Mn1.5 O4 oxide as an example. The modified oxide delivers extraordinarily stable battery performance, achieving 71.4 % capacity retention after 2000 cycles at 1 C. This work shows that an orbital-level understanding can be leveraged to engineer high structural stability of the anion oxygen framework of oxides. Moreover, the similarity of the oxygen lattice between oxide electrodes makes this approach extendable to other electrodes, with orbital-focused engineering a new avenue for the fundamental modification of battery materials.


Expression of heparanase in renal cell carcinomas: implications for tumor invasion and prognosis.
Heparanase activity has been detected in many malignant tumors, showing a correlation with the metastatic potential. The present study was undertaken to investigate the expression of heparanase and its prognostic significance in renal cell carcinomas (RCC). Nineteen RCCs and 6 nonneoplastic renal tissues were analyzed for heparanase mRNA expression by real-time PCR. Heparanase protein expression was semiquantitatively investigated by immunohistochemistry in 70 RCCs. Involvement of heparanase in the invasiveness of RCC cell lines, 786-O and Caki-2 cells, was examined by down-regulating the gene expression with small interfering RNA (siRNA) using the Matrigel invasion assay. The expression level of heparanase mRNA was significantly higher in clear cell RCCs than in papillary RCCs, chromophobe RCCs, and nonneoplastic renal tissues. Heparanase was predominantly immunolocalized to cell surface and cytoplasm of clear cell RCCs and mean expression levels of heparanase were significantly higher in clear cell RCCs than in papillary and chromophobe RCCs. The protein expression levels were positively correlated with primary tumor stage, distant metastasis, and histologic grade. Targeting of heparanase mRNA expression in 786-O and Caki-2 cells with siRNA down-regulated the mRNA expression and inhibited the Matrigel invasion by these cells, whereas nonsilencing siRNA showed no effect. Multivariate Cox analysis revealed that elevated heparanase expression was a significant and an independent predictor of disease-specific survival (odds ratio, 8.814; P = 0.019). These data suggest that heparanase plays an important role in invasion and metastasis and silencing of the gene might be a potential therapeutic target in clear cell RCCs.


In vitro survival and differentiation of neurons derived from epidermal growth factor-responsive postnatal hippocampal stem cells: inducing effects of brain-derived neurotrophic factor.
Neural stem cells proliferate in vitro and form neurospheres in the presence of epidermal growth factor (EGF), and are capable of differentiating into both neurons and glia when exposed to a substrate. We hypothesize that specific neurotrophic factors induce differentiation of stem cells from different central nervous system (CNS) regions into particular fates. We investigated differentiation of stem cells from the postnatal mouse hippocampus in culture using the following trophic factors (20 ng/mL): brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3) and glial-derived neurotrophic factor (GDNF). Without trophic factors, 32% of stem cells differentiated into neurons by 4 days in vitro (DIV), decreasing to 10% by 14 DIV. Addition of BDNF (starting at either day 0 or day 3) significantly increased neuron survival (31-43% by 14 DIV) and differentiation. Morphologically, many well-differentiated neurons resembled hippocampal pyramidal neurons. 5'-Bromodeoxyuridine labeling demonstrated that the pyramidal-like neurons originated from stem cells which had proliferated in EGF-containing cultures. However, similar application of NT-3 and GDNF did not exert such a differentiating effect. Addition of BDNF to stem cells from the postnatal cerebellum, midbrain, and striatum did not induce these neuronal phenotypes, though similar application to cortical stem cells yielded pyramidal-like neurons. Thus, BDNF supports survival of hippocampal stem cell-derived neurons and also can induce differentiation of these cells into pyramidal-like neurons. The presence of pyramidal neurons in BDNF-treated hippocampal and cortical stem cell cultures, but not in striatal, cerebellar, and midbrain stem cell cultures, suggests that stem cells from different CNS regions differentiate into region-specific phenotypic neurons when stimulated with an appropriate neurotrophic factor.


Impairment of learning and memory after photothrombosis of the prefrontal cortex in rat brain: effects of Noopept.
Experiments were performed on rats trained conditioned passive avoidance response. Acquisition and retention of memory traces were impaired after photothrombosis of the prefrontal cortex. The acyl-prolyl-containing dipeptide Noopept facilitated retention and retrieval of a conditioned passive avoidance response, normalized learning capacity in animals with ischemic damage to the cerebral cortex, and promoted finish training in rats with hereditary learning deficit. These results show that Noopept improves all three stages of memory. It should be emphasized that the effect of Noopept was most pronounced in animals with impaired mnesic function.


Novel germanium/polypyrrole composite for high power lithium-ion batteries.
Nano-Germanium/polypyrrole composite has been synthesized by chemical reduction method in aqueous solution. The Ge nanoparticles were directly coated on the surface of the polypyrrole. The morphology and structural properties of samples were determined by X-ray diffraction, scanning electron microscopy and transmission electron microscopy. Thermogravimetric analysis was carried out to determine the polypyrrole content. The electrochemical properties of the samples have been investigated and their suitability as anode materials for the lithium-ion battery was examined. The discharge capacity of the Ge nanoparticles calculated in the Ge-polypyrrole composite is 1014 mAh g(-1) after 50 cycles at 0.2 C rate, which is much higher than that of pristine germanium (439 mAh g(-1)). The composite also demonstrates high specific discharge capacities at different current rates (1318, 1032, 661, and 460 mAh g(-1) at 0.5, 1.0, 2.0, and 4.0 C, respectively). The superior electrochemical performance of Ge-polypyrrole composite could be attributed to the polypyrrole core, which provides an efficient transport pathway for electrons. SEM images of the electrodes have demonstrated that polypyrrole can also act as a conductive binder and alleviate the pulverization of electrode caused by the huge volume changes of the nanosized germanium particles during Li(+) intercalation/de-intercalation.


Calmodulin-dependent glycogen synthase kinase: identification in liver of normal and phosphorylase kinase-deficient rats.
We have purified a calmodulin-dependent glycogen synthase kinase from livers of normal and phosphorylase kinase-deficient (gsd/gsd) rats. No differences between normal and gsd/gsd rats were apparent in either (a) the ability of liver extracts to phosphorylate exogenous glycogen synthase in a Ca2+- and calmodulin-dependent manner or (b) the purification of the calmodulin-dependent synthase kinase. Although extracts from rat liver, when compared to rabbit liver extracts, had a significantly reduced ability to phosphorylate exogenous synthase, the calmodulin-dependent synthase kinase could be purified from rat liver using a protocol identical to that described for rabbit liver. Moreover, the synthase kinase purified from rat liver had properties very similar to those of the rabbit liver enzyme. The enzyme was completely dependent on calmodulin for activity against glycogen synthase, was unable to phosphorylate phosphorylase b, catalyzed the rapid incorporation of 0.4 mol phosphate/mol of glycogen synthase subunit, selectively phosphorylated sites 1b and 2 in the glycogen synthase molecule, had a Stokes' radius of about 70 A, and appeared to be composed of subunits of Mr 56,000 and 57,000. These observations led us to conclude that (1) calmodulin-dependent glycogen synthase kinase is distinct from other kinases previously described and (2) the rat liver kinase and the rabbit liver kinase are very similar enzymes.


The effects of fluoride on cell growth of two human cell lines and on DNA and protein synthesis in HeLa cells.
The effect of sodium fluoride on the growth of two continuous human cell lines, i.e. HeLa cells and human conjunctiva clone 1-5C-4 cells, was studied. The growth of HeLa cells and clone 1-5C-4 cells was arrested nearly completely by the addition of 0.95 and 1,90 mM of sodium fluoride, respectively. DNA synthesis in HeLa cells, determined by incorporation of 3H-thymidine, was not affected appreciably for the first 24-hr period after the addition of sodium fluoride. Markedly reduced incorporation, however, occurred during the next 24-hr period. Thus, there was a discrepancy between the immediate cessation of cell division and the delayed suppression of DNA synthesis. On the other hand, a suppressive effect of sodium fluoride on protein synthesis determined by 14C-leucine incorporation was evident already during the first 24-hr period. The results indicate that the inhibition of protein synthesis is the main cause of growth inhibition.


Certified safe farm: identifying and removing hazards on the farm.
This article describes the development of the Certified Safe Farm (CSF) on-farm safety review tools, characterizes the safety improvements among participating farms during the study period, and evaluates differences in background variables between low and high scoring farms. Average farm review scores on 185 study farms improved from 82 to 96 during the five-year study (0-100 scale, 85 required for CSF certification). A total of 1292 safety improvements were reported at an estimated cost of $650 per farm. A wide range of improvements were made, including adding 9 rollover protective structures (ROPS), 59 power take-off (PTO) master shields, and 207 slow-moving vehicle (SMV) emblems; improving lighting on 72 machines: placing 171 warning decals on machinery; shielding 77 moving parts; locking up 17 chemical storage areas, adding 83 lockout/tagout improvements; and making general housekeeping upgrades in 62 farm buildings. The local, trained farm reviewers and the CSF review process overall were well received by participating farmers. In addition to our earlier findings where higher farm review scores were associated with lower self-reported health outcome costs, we found that those with higher farm work hours, younger age, pork production in confinement, beef production, poultry production, and reported exposure to agrichemicals had higher farm review scores than those who did not have these characteristics. Overall, the farm review process functioned as expected. encouraging physical improvements in the farm environment, and contributing to the multi-faceted CSF intervention program.


Conserved cobalamin acquisition protein 1 is essential for vitamin B12 uptake in both Chlamydomonas and Phaeodactylum.
Microalgae play an essential role in global net primary productivity and global biogeochemical cycling. Despite their phototrophic lifestyle, over half of algal species depend for growth on acquiring an external supply of the corrinoid vitamin B12 (cobalamin), a micronutrient produced only by a subset of prokaryotic organisms. Previous studies have identified protein components involved in vitamin B12 uptake in bacterial species and humans. However, little is known about its uptake in algae. Here, we demonstrate the essential role of a protein, cobalamin acquisition protein 1 (CBA1), in B12 uptake in Phaeodactylum tricornutum using CRISPR-Cas9 to generate targeted knockouts and in Chlamydomonas reinhardtii by insertional mutagenesis. In both cases, CBA1 knockout lines could not take up exogenous vitamin B12. Complementation of the C. reinhardtii mutants with the wild-type CBA1 gene restored B12 uptake, and regulation of CBA1 expression via a riboswitch element enabled control of the phenotype. When visualised by confocal microscopy, a YFP-fusion with C. reinhardtii CBA1 showed association with membranes. Bioinformatics analysis found that CBA1-like sequences are present in all major eukaryotic phyla. In algal taxa, the majority that encoded CBA1 also had genes for B12-dependent enzymes, suggesting CBA1 plays a conserved role. Our results thus provide insight into the molecular basis of algal B12 acquisition, a process that likely underpins many interactions in aquatic microbial communities.


Enhanced primary treatment of concentrated black water and kitchen residues within DESAR concept using two types of anaerobic digesters.
Anaerobic digestion of concentrated domestic wastewater streams--black or brown water, and solid fraction of kitchen waste is considered as a core technology in a source separation based sanitation concept (DESAR--decentralised sanitation and reuse). A simple anaerobic digester can be implemented for an enhanced primary treatment or, in some situations, as a main treatment. Two reactor configurations were extensively studied; accumulation system (AC) and UASB septic tank at 15, 20 and 25 degrees C. Due to long retention times in an AC reactor, far stabilisation of treated medium can be accomplished with methanisation up to 60%. The AC systems are the most suitable to apply when the volume of waste to be treated is minimal and when a direct reuse of a treated medium in agriculture is possible. Digested effluent contains both liquid and solids. In a UASB septic tank, efficient separation of solids and liquid is accomplished. The total COD removal was above 80% at 25 degrees C. The effluent contains COD and nutrients, mainly in a soluble form. The frequency of excess sludge removal is low and sludge is well stabilised due to a long accumulation time.


MTERF4 regulates translation by targeting the methyltransferase NSUN4 to the mammalian mitochondrial ribosome.
Precise control of mitochondrial DNA gene expression is critical for regulation of oxidative phosphorylation capacity in mammals. The MTERF protein family plays a key role in this process, and its members have been implicated in regulation of transcription initiation and site-specific transcription termination. We now demonstrate that a member of this family, MTERF4, directly controls mitochondrial ribosomal biogenesis and translation. MTERF4 forms a stoichiometric complex with the ribosomal RNA methyltransferase NSUN4 and is necessary for recruitment of this factor to the large ribosomal subunit. Loss of MTERF4 leads to defective ribosomal assembly and a drastic reduction in translation. Our results thus show that MTERF4 is an important regulator of translation in mammalian mitochondria.


Parkin is a lipid-responsive regulator of fat uptake in mice and mutant human cells.
It has long been hypothesized that abnormalities in lipid biology contribute to degenerative brain diseases. Consistent with this, emerging epidemiologic evidence links lipid alterations with Parkinson disease (PD), and disruption of lipid metabolism has been found to predispose to α-synuclein toxicity. We therefore investigated whether Parkin, an E3 ubiquitin ligase found to be defective in patients with early onset PD, regulates systemic lipid metabolism. We perturbed lipid levels by exposing Parkin+/+ and Parkin-/- mice to a high-fat and -cholesterol diet (HFD). Parkin-/- mice resisted weight gain, steatohepatitis, and insulin resistance. In wild-type mice, the HFD markedly increased hepatic Parkin levels in parallel with lipid transport proteins, including CD36, Sr-B1, and FABP. These lipid transport proteins were not induced in Parkin-/- mice. The role of Parkin in fat uptake was confirmed by increased oleate accumulation in hepatocytes overexpressing Parkin and decreased uptake in Parkin-/- mouse embryonic fibroblasts and patient cells harboring complex heterozygous mutations in the Parkin-encoding gene PARK2. Parkin conferred this effect, in part, via ubiquitin-mediated stabilization of the lipid transporter CD36. Reconstitution of Parkin restored hepatic fat uptake and CD36 levels in Parkin-/- mice, and Parkin augmented fat accumulation during adipocyte differentiation. These results demonstrate that Parkin is regulated in a lipid-dependent manner and modulates systemic fat uptake via ubiquitin ligase-dependent effects. Whether this metabolic regulation contributes to premature Parkinsonism warrants investigation.


Reinitiation of growth in senescent mouse mammary epithelium in response to cholera toxin.
Several lines of mouse mammary tissue that had been serially transplanted until mitotic senescence was reached were exposed in vivo to plastic implants that slowly released cholera toxin. Gland tissue surrounding the implants displayed new end buds, indicating reinitiation of growth and morphogenesis. The ability of cholera toxin, which elevates intracellular adenosine 3',5'-monophosphate, to temporarily reverse the senescent phenotype suggests that this mitotic dysfunction results not from generalized cellular deterioration but from specific changes in cell regulation.


The male sterile 8 mutation of maize disrupts the temporal progression of the transcriptome and results in the mis-regulation of metabolic functions.
Maize anther ontogeny is complex, with the expression of more than 30,000 genes over 4 days of cell proliferation, cell fate acquisition and the start of meiosis. Although many male-sterile mutants disrupt these key steps, few have been investigated in detail. The terminal phenotypes of Zea mays (maize) male sterile 8 (ms8) are small anthers exhibiting meiotic failure. Here, we document much earlier defects: ms8 epidermal cells are normal in number but fail to elongate, and there are fewer, larger tapetal cells that retain, rather than secrete, their contents. ms8 meiocytes separate early, have extra space between them, occupied by excess callose, and the meiotic dyads abort. Thousands of transcriptome changes occur in ms8, including ectopic activation of genes not expressed in fertile siblings, failure to express some genes, differential expression compared with fertile siblings and about 40% of the differentially expressed transcripts appear precociously. There is a high correlation between mRNA accumulation assessed by microarray hybridization and quantitative real-time reverse transcriptase polymerase chain reaction. Sixty-three differentially expressed proteins were identified after two-dimensional gel electrophoresis followed by liquid chromatography tandem mass spectroscopy, including those involved in metabolism, plasmodesmatal remodeling and cell division. The majority of these were not identified by differential RNA expression, demonstrating the importance of proteomics in defining developmental mutants.


Autotransfusion of drained mediastinal blood.
The Sorenson equipment for transfusion of drained mediastinal blood has been widely used in the United States. We have tested its use postoperatively in 10 patients, assessing its ease of use from the medical and nursing views as well as analyzing the blood constituents which would be infused into the patients. The hemoglobin content of the infused drainage averaged 6 g/dl compared with 12 g/dl in the patients' blood. Platelet counts were very low, averaging 15 X 10(9)/l compared with levels of 110 X 10(9)/l for the patient. Platelet aggregation in response to ADP stimuli was assessed and was grossly deficient in the collected blood showing that the platelets that were present were not capable of aggregating. Protein and electrolyte levels were substantially the same in both series of specimens. Handling difficulties with the equipment together with these rather disappointing hematological results suggest that the equipment is only useful where excessive postoperative bleeding is encountered or where there is particular need to conserve red blood cells.


Peroxisomal compartmentalization of amino acid biosynthesis reactions imposes an upper limit on compartment size.
Cellular metabolism relies on just a few redox cofactors. Selective compartmentalization may prevent competition between metabolic reactions requiring the same cofactor. Is such compartmentalization necessary for optimal cell function? Is there an optimal compartment size? Here we probe these fundamental questions using peroxisomal compartmentalization of the last steps of lysine and histidine biosynthesis in the fission yeast Schizosaccharomyces japonicus. We show that compartmentalization of these NAD+ dependent reactions together with a dedicated NADH/NAD+ recycling enzyme supports optimal growth when an increased demand for anabolic reactions taxes cellular redox balance. In turn, compartmentalization constrains the size of individual organelles, with larger peroxisomes accumulating all the required enzymes but unable to support both biosynthetic reactions at the same time. Our reengineering and physiological experiments indicate that compartmentalized biosynthetic reactions are sensitive to the size of the compartment, likely due to scaling-dependent changes within the system, such as enzyme packing density.


Postfamine stature and socioeconomic status in Ireland.
Previous research has documented socioeconomic stratification of secular trend in height in historical populations. Using data from 4,900 males and 1,430 females born between 1840 and 1910 collected as part of the Harvard Anthropological Survey of Ireland (1934-1936), this study examined the secular changes in postfamine Ireland using several socioeconomic variables, including: occupation, migration, education, siblings, birthplace, and occupation of father and mother's father. Correlations were also calculated between height and various historical economic indices. Significant differences in the height of Irish males were found by occupation, education, and socioeconomic status of father and maternal grandfather. Males employed in agriculture, or whose fathers or grandfathers were so employed, were significantly taller than other males. For the smaller female sample, only occupation and grandfather's socioeconomic status had a significant impact on height. An inverse correlation was also found between the British Cost of Living Index (BCL) and male heights. Our results suggest that availability of resources plays an important role in the overall nutritional status reflected in terminal adult height.


Effect of amplitude modulation on selective excitation in NMR imaging.
If NMR imaging uses selective excitation, then it has to do it correctly as it affects the image resolution. We have related the properties of the selected slice, such as its width, its rate of attenuation, and its side lobes to the amplitude modulation of the radio frequency pulse. An interesting observation was that multiplying sinc(t) function by a triangle can give better results than multiplying it with a Gauss function. We have used computer simulation and tried to emphasize the qualitative relationship between the properties. We conclude with some practical requirements and show the need of a numerical design procedure if the slice properties have to be optimized.


Depolarization of polymorphonuclear leukocytes by Porphyromonas (Bacteroides) gingivalis 381 in the absence of respiratory burst activation.
Bacteroides spp. may contribute to the chronicity of mixed infections by affecting the normal functions of polymorphonuclear leukocytes (PMN). This study evaluated the physiologic and biochemical responses of human peripheral blood PMN to a variety of strains of the oral periodontal pathogen Porphyromonas (Bacteroides) gingivalis. Strain 381 and ATCC type strain 33277 caused rapid and lasting depolarization of the electrochemical potential that exists across the PMN membrane by a mechanism that was independent of activation of the pertussis toxin-sensitive G protein or protein kinase C. Membrane depolarization did not initiate increases in intracellular calcium or respiratory burst activation, and activity was not inhibited by surface proteolysis or sugars. However, membrane depolarization was associated with inhibition of PMN responses to the chemotactic peptide N-formylmethionyl leucyl phenylalanine. Membrane-depolarizing activity was isolated with the outer membrane of strain 381 by surface extraction of the bacteria by using Zwittergent 3,14, followed by Sephacryl S-200 gel filtration chromatography. The partially purified outer membrane components were heat stable, were not inhibited by tosyl-lysine chloromethyl ketone, and inhibited N-formylmethionyl leucyl phenylalanine-stimulated superoxide production. The results suggest that outer membrane components of P. gingivalis 381 and 33277 have porinlike activity that can depolarize PMN membranes and immobilize PMN responses to chemotactic peptides. This may prove to be an important virulence characteristic of these strains.


Cadmium uptake and zinc-cadmium antagonism in Australian tropical rock oysters: Potential solutions for oyster aquaculture enterprises.
Variable and occasionally high concentrations of cadmium in wild oysters at a remote location with the potential to develop aquaculture enterprises motivated research into the distribution and sources of metals in oysters, seawater, sediment, suspended solids and phytoplankton. Saccostrea mytiloides and Saccostrea mordax contained cadmium concentrations exceeding the food standard maximum level (ML) at three of four sites. At one site with high zinc levels in sediment, oyster cadmium levels were below the ML. Metal levels in seawater were not correlated with cadmium levels in oysters but high cadmium/zinc ratios were measured in Trichodesmium erythraeum blooms. We suggest that oysters accumulate cadmium mainly from annual phytoplankton blooms except at sites where zinc availability is sufficiently high to prevent uptake though a mechanism of antagonistic exclusion. Knowledge of the source and uptake mechanisms of cadmium in oysters should lead to new management strategies to reduce cadmium levels in farmed oysters.