Microtubule-Dependent Mitochondria Alignment Regulates Calcium Release in Response to Nanomechanical Stimulus in Heart Myocytes.
Arrhythmogenesis during heart failure is a major clinical problem. Regional electrical gradients produce arrhythmias, and cellular ionic transmembrane gradients are its originators. We investigated whether the nanoscale mechanosensitive properties of cardiomyocytes from failing hearts have a bearing upon the initiation of abnormal electrical activity. Hydrojets through a nanopipette indent specific locations on the sarcolemma and initiate intracellular calcium release in both healthy and heart failure cardiomyocytes, as well as in human failing cardiomyocytes. In healthy cells, calcium is locally confined, whereas in failing cardiomyocytes, calcium propagates. Heart failure progressively stiffens the membrane and displaces sub-sarcolemmal mitochondria. Colchicine in healthy cells mimics the failing condition by stiffening the cells, disrupting microtubules, shifting mitochondria, and causing calcium release. Uncoupling the mitochondrial proton gradient abolished calcium initiation in both failing and colchicine-treated cells. We propose the disruption of microtubule-dependent mitochondrial mechanosensor microdomains as a mechanism for abnormal calcium release in failing heart.


Neurotoxic potential of reactive astrocytes in canine distemper demyelinating leukoencephalitis.
Canine distemper virus (CDV) causes a fatal demyelinating leukoencephalitis in young dogs resembling human multiple sclerosis. Astrocytes are the main cellular target of CDV and undergo reactive changes already in pre-demyelinating brain lesions. Based on their broad range of beneficial and detrimental effects in the injured brain reactive astrogliosis is in need of intensive investigation. The aim of the study was to characterize astrocyte plasticity during the course of CDV-induced demyelinating leukoencephalitis by the aid of immunohistochemistry, immunofluorescence and gene expression analysis. Immunohistochemistry revealed the presence of reactive glial fibrillary acidic protein (GFAP)+ astrocytes with increased survivin and reduced aquaporin 4, and glutamine synthetase protein levels, indicating disturbed blood brain barrier function, glutamate homeostasis and astrocyte maladaptation, respectively. Gene expression analysis revealed 81 differentially expressed astrocyte-related genes with a dominance of genes associated with neurotoxic A1-polarized astrocytes. Accordingly, acyl-coA synthetase long-chain family member 5+/GFAP+, and serglycin+/GFAP+ cells, characteristic of A1-astrocytes, were found in demyelinating lesions by immunofluorescence. In addition, gene expression revealed a dysregulation of astrocytic function including disturbed glutamate homeostasis and altered immune function. Observed findings indicate an astrocyte polarization towards a neurotoxic phenotype likely contributing to lesion initiation and progression in canine distemper leukoencephalitis.


Relationships between absorption efficiency of elements in mammals and chemical properties.
Oral absorption efficiency is an important factor to consider in human risk assessment and varies widely between elements. Linking absorption efficiency to chemical properties facilitates the understanding of underlying processes and enables extrapolation across elements. In our study, oral absorption efficiency in humans was predicted for a number of elements based on their ionization energy and electronegativity. Data on oral absorption efficiency in humans were retrieved via a literature survey. A model was developed based on the assumption that ionic species readily react with biotic ligands. Accordingly, ionization energy was presumed to represent the reactivity and absorption of atoms in the gastrointestinal tract. The coefficients of the model were parameterized by fitting the quantitative relationship between absorption efficiency and ionization energy to data collected from well-standardized studies. Generally, absorption efficiency was strongly related to ionization energy, explaining 94% of the variability in absorption efficiency between elements reported by the International Commission on Radiological Protection (ICRP). In addition, the absorption efficiencies predicted based on ionization energy were within a factor of two of those given by the ICRP (ME = -0.05; RMSE = 0.31). However, the model is not applicable to alkaline metals and molybdenum because of the uniquely high solubility of their compounds or the flexible electron configuration of these elements. Approximately 56% of the variability in absorption efficiency between elements could be explained by electronegativity. These strong relationships between absorption efficiency and ionization energy and, to a lesser extent, electronegativity indicate potential for extrapolation across elements using atomic properties.


Genetic modifiers of cardiorespiratory fitness response to lifestyle intervention.
Numerous prospective studies indicate that improved cardiorespiratory fitness reduces type 2 diabetes risk and delays disease progression. We hypothesized that genetic variants modify fitness response to an intensive lifestyle intervention (ILI) in the Action for Health in Diabetes (Look AHEAD) randomized clinical trial, aimed to detect whether ILI will reduce cardiovascular events in overweight/obese subjects with type 2 diabetes compared with a standard of care. Polymorphisms in established fitness genes and in all loci assayed on the Illumina CARe iSelect chip were examined as predictors of change in MET level, estimated using a treadmill test, in response to a 1-yr intervention in 3899 participants. We identified a significant signal in previously reported fitness-related gene RUNX1 that was associated with 1-yr METs response in ILI (0.19 ± 0.04 MET less improvement per minor allele copy; P = 1.9 × 10(-5)) and genotype-intervention interaction (P = 4.8 × 10(-3)). In the chipwide analysis, FKBP7 rs17225700 showed a significant association with ILI response among subjects not receiving beta-blocker medications (0.47 ± 0.09 METs less improvement; P = 5.3 × 10(-5)) and genotype-treatment interaction (P = 5.3 × 10(-7)). The Gene Relationships Among Implicated Loci pathway-based analysis identified connections between associated genes, including those influencing vascular tone, muscle contraction, cardiac energy substrate dynamics, and muscle protein synthesis. This is the first study to identify genetic variants associated with fitness responses to a randomized lifestyle intervention in overweight/obese diabetic individuals. RUNX1 and FKBP7, involved in erythropoesis and muscle protein synthesis, respectively, are related to change in cardiorespiratory fitness in response to exercise.


Prostate cancer localization by novel magnetic resonance dispersion imaging.
Diagnosis and focal treatment of prostate cancer, the most prevalent form of cancer in men, is hampered by the limits of current clinical imaging. Angiogenesis imaging is a promising option for detection and localization of prostate cancer. It can be imaged by dynamic contrast-enhanced (DCE) MRI, assessing microvascular permeability as an indicator for angiogenesis. However, information on microvascular architecture changes associated with angiogenesis is not available. This paper presents a new model enabling the combined assessment of microvascular permeability and architecture. After the intravenous injection of a gadolinium-chelate bolus, time-concentration curves (TCCs) are measured by DCE-MRI at each voxel. According to the convective dispersion equation, the microvascular architecture is reflected in the dispersion coefficient. A solution of this equation is therefore proposed to represent the intravascular blood plasma compartment in the Tofts model. Fitting the resulting model to TCCs measured at each voxel leads to the simultaneous generation of a dispersion and a permeability map. Measurement of an arterial input function is no longer required. Preliminary validation was performed by spatial comparison with the histological results in seven patients referred for radical prostatectomy. Cancer localization by the obtained dispersion maps provided an area under the receiver operating characteristic curve equal to 0.91. None of the standard DCE-MRI parametric maps could outperform this result, motivating towards an extended validation of the method, also aimed at investigating other forms of cancer with pronounced angiogenic development.


Bark flammability as a fire-response trait for subalpine trees.
Relationships between the flammability properties of a given plant and its chances of survival after a fire still remain unknown. We hypothesize that the bark flammability of a tree reduces the potential for tree survival following surface fires, and that if tree resistance to fire is provided by a thick insulating bark, the latter must be few flammable. We test, on subalpine tree species, the relationship between the flammability of bark and its insulating ability, identifies the biological traits that determine bark flammability, and assesses their relative susceptibility to surface fires from their bark properties. The experimental set of burning properties was analyzed by Principal Component Analysis to assess the bark flammability. Bark insulating ability was expressed by the critical time to cambium kill computed from bark thickness. Log-linear regressions indicated that bark flammability varies with the bark thickness and the density of wood under bark and that the most flammable barks have poor insulating ability. Susceptibility to surface fires increases from gymnosperm to angiosperm subalpine trees. The co-dominant subalpine species Larix decidua (Mill.) and Pinus cembra (L.) exhibit large differences in both flammability and insulating ability of the bark that should partly explain their contrasted responses to fires in the past.


Segmentation of the Himalayas as revealed by arc-parallel gravity anomalies.
Lateral variations along the Himalayan arc are suggested by an increasing number of studies and carry important information about the orogen's segmentation. Here we compile the hitherto most complete land gravity dataset in the region which enables the currently highest resolution plausible analysis. To study lateral variations in collisional structure we compute arc-parallel gravity anomalies (APaGA) by subtracting the average arc-perpendicular profile from our dataset; we compute likewise for topography (APaTA). We find no direct correlation between APaGA, APaTA and background seismicity, as suggested in oceanic subduction context. In the Himalayas APaTA mainly reflect relief and erosional effects, whereas APaGA reflect the deep structure of the orogen with clear lateral boundaries. Four segments are outlined and have disparate flexural geometry: NE India, Bhutan, Nepal &India until Dehradun, and NW India. The segment boundaries in the India plate are related to inherited structures, and the boundaries of the Shillong block are highlighted by seismic activity. We find that large earthquakes of the past millennium do not propagate across the segment boundaries defined by APaGA, therefore these seem to set limits for potential rupture of megathrust earthquakes.


Neuropathic Pain.
Neuropathic pain is a frequently encountered condition that is often resistant to treatment and is associated with poor patient satisfaction of their treatment. Several medications have been shown to be effective in treating neuropathic pain associated with diabetic neuropathy and postherpetic neuralgia, and these medications are often used to treat neuropathic pain associated with other conditions as well. This article summarizes the diagnosis and assessment of patients with neuropathic pain as well as available pharmacologic and interventional treatment options. Evidence-based recommendations for the treatment of neuropathic pain have been published, and first-line medications include antidepressants, anticonvulsants, topical agents, as well as opioid analgesics. Interventional options include anesthetic and steroid injections, nerve blocks, and spinal cord stimulation. Essential to the treatment algorithm of neuropathic pain is the assessment and treatment of psychosocial comorbidities and the utilization of a multidisciplinary team approach, including cognitive-behavioral and rehabilitative therapies. Questions remain about the comparative effectiveness of various medications and combination therapies. Increasing interest also exists in the optimization and personalization of pharmacotherapy based upon the underlying mechanism(s) of neuropathic pain according to the quality of the patient's symptoms. The management of chronic neuropathic pain is challenging and is best achieved with the use of a multidisciplinary team. Pain is a subjective experience, and it is important to validate a patient's pain, address psychosocial comorbidities, and set realistic treatment goals. Evidence-based guidelines are available to guide treatment, but frequently, high-quality evidence-based recommendations are lacking.


Aminoglycoside resistance and aminoglycoside usage: ten years of experience in one hospital.
For 10 years the 700-bed Minneapolis Veterans Affairs Medical Center has conducted a policy of carefully controlled aminoglycoside usage and monitoring of resistance of over 25,000 aerobic and facultative gram-negative bacillary isolates to the aminoglycosides. On two occasions during the 1980s, our experience of introducing amikacin at a high level of usage was associated with a significant reduction in resistance to gentamicin and tobramycin among gram-negative bacilli. Rapid reintroduction of gentamicin usage in 1982 after the first amikacin period was associated with a significant and rapid increase in gentamicin and tobramycin resistance. However, in 1986, gentamicin was again reintroduced to this institution at an initially modest level, and the percentage of usage of gentamicin was gradually increased over a 15-month period without a significant change in resistance to gentamicin, tobramycin, or amikacin while maintaining an overall 68% gentamicin usage and 30% amikacin usage. Aminoglycoside usage (measured as patient days) rose steadily from under 2,000 patient days per quarter in 1980 and 1981 to over 3,000 days per quarter in 1985. Since 1985, usage has declined to under 2,500 patient days per quarter in 1990. This usage rise and fall occurred during a steadily declining daily patient census that was 590 in 1980 and 465 in 1989. A move to a new hospital building in June 1988 was associated with an additional significant decline in resistance to all aminoglycosides (P less than 0.05), continuing a trend that was evident for the year preceding the move. Resistance to aminoglycoside antibiotics is now at the lowest level in 10 years at this institution, with only one gram-negative organism, Pseudomonas aeruginosa, that exhibits more than 5% resistance to gentamicin and no gram-negative species that are more than 5% resistant to amikacin and tobramycin.


Phencyclidine (PCP)-induced deficits of prepulse inhibition in monkeys.
Prepulse inhibition (PPI) of the acoustic startle reflex is a measure of sensorimotor gating which occurs in both rodents and humans. PPI is deficient in severe neuropsychiatric disorders such as schizophrenia. We investigated PPI in 10 adult monkeys (Cebus apella). Stimuli were 115 dB white noise startle pulses, either alone or preceded by 120 ms with a prepulse of either 8 or 16 dB above the 70 dB background noise. Experiments included a pretreatment baseline session and a session following treatment with either phencyclidine (PCP, 0.12 mg/kg, i.m.) or saline. Comparison of peak amplitudes indicated a significant intensity-dependent decrease in startle response that was similar to that observed in humans under similar experimental conditions. PCP treatment significantly disrupted PPI, but did not reduce responses to startle pulses alone. These results provide the first demonstration of PPI in monkeys. The ability of PCP to induce schizophrenia-like deficits in PPI suggests that PPI in nonhuman primates may provide an important animal model for the development of novel anti-schizophrenia medications.


A manufacturer's perspective: Hewlett Packard Y2K action plan.
Medical device manufacturers must ensure that their devices are safe and effective including investigating issues involved with the century rollover. Manufacturers must begin early to evaluate their products in order to allow time to correct and distribute these product corrections and communicate to their customers so they can prepare for the Y2K event.


Inositol phosphate release and steroidogenesis in rat adrenal glomerulosa cells. Comparison of the effects of endothelin, angiotensin II and vasopressin.
Endothelin has steroidogenic activity in adrenal glomerulosa cells, as do two other vasoconstrictor peptides, angiotensin II and vasopressin. The steroidogenic activities of angiotensin II and vasopressin are probably mediated via the phosphatidylinositol-turnover pathway and associated changes in cytosolic Ca2+ concentration. Endothelin caused a steroidogenic response, which was small compared with that to angiotensin II and quantitatively similar to the vasopressin response. Cytosolic free Ca2+ responses were similarly higher to angiotensin II than to either of the other two peptides. However, total inositol phosphate responses to endothelin and angiotensin II were similar when these were measured over 20 min, and were quantitatively greater than the vasopressin response. A detailed study has been made of the phosphatidylinositol-turnover response to endothelin in comparison with responses to angiotensin II and vasopressin. Each of the three peptides produced a rapid and transient rise in Ins(1,4,5)P3 (max. 5-15 s), followed by a slow sustained rise. Ins(1,4,5)P3 was metabolized by both dephosphorylation and phosphorylation pathways, but the relative importance of the two metabolic pathways was different under stimulation by each of the three peptides. These findings show that adrenal glomerulosa cells can distinguish between the stimulation of phosphatidylinositol turnover by three different effectors. These differences in the pathway may be associated with the observed different steroidogenic and Ca2+ responses to the three peptides.


Bcl-xL regulates CD1d-mediated antigen presentation to NKT cells by altering CD1d trafficking through the endocytic pathway.
NKT cells are a unique subset of T cells that recognize glycolipid Ags presented in the context of CD1d molecules. NKT cells mount strong antitumor responses and are a major focus in developing effective cancer immunotherapy. It is known that CD1d molecules are constantly internalized from the cell surface, recycled through the endocytic compartments, and re-expressed on the cell surface. However, little is known about the regulation of CD1d-mediated Ag processing and presentation in B cell lymphoma. Prosurvival factors of the Bcl-2 family, such as Bcl-xL, are often upregulated in B cell lymphomas and are intimately linked to sphingolipid metabolism, as well as the endocytic compartments. We hypothesized that Bcl-xL can regulate CD1d-mediated Ag presentation to NKT cells. We found that overexpression or induction of Bcl-xL led to increased Ag presentation to NKT cells. Conversely, the inhibition or knockdown of Bcl-xL led to decreased NKT cell activation. Furthermore, knockdown of Bcl-xL resulted in the loss of CD1d trafficking to lysosome-associated membrane protein 1(+) compartments. Rab7, a late endosomal protein, was upregulated and CD1d molecules accumulated in the Rab7(+) late endosomal compartment. These results demonstrate that Bcl-xL regulates CD1d-mediated Ag processing and presentation to NKT cells by altering the late endosomal compartment and changing the intracellular localization of CD1d.


Fat distribution of overweight persons in relation to morbidity and subjective health.
The association between fat distribution, morbidity and subjective health was studied in 95 overweight adult men and 210 overweight adult women. Retrospective morbidity data were taken from a continuous morbidity registration made by general practitioners over a period of maximally 17 years. In addition information about subjective health and weight history was obtained from a self-administered questionnaire. Anthropometric measurements were taken and, on the basis of waist-hip and waist-thigh circumference ratios, subjects were classified into upper body segment obesity, intermediate obesity, and lower body segment obesity. It was found that, adjusted for age and body mass index, a high waist-thigh circumference ratio was a risk factor for hypertension and for gout or diabetes in women and arthrosis in men. A low waist-thigh ratio was associated with a high prevalence of varicose veins in women. The associations of waist-hips circumference ratio with morbidity were less pronounced, with the exception of hypertension in men. Information from the questionnaire revealed that persons with upper body segment obesity (especially men) felt less healthy and had more health complaints. These findings were more pronounced for subjects less than 50 years of age than for those of 50 years and older. The weight histories suggest that women with lower body segment obesity had a longer history of obesity than women with upper body segment obesity. This was not found in men. It is concluded that classification of obesity on the basis of circumference ratios is useful for the evaluation of health hazards of overweight subjects.


Evaluation of pediatric use patterns and performance of lap shoulder belt systems in the center rear.
Lap and shoulder belts have been required in rear outboard positions since 1989. A recent congressional mandate encouraged the requirement of a lap and shoulder belt in the center rear seat position. This study utilized Data from the Partners for Child Passenger Safety study, a large-scale child-focused crash surveillance system, to quantify changes in seating patterns for children in vehicles that already have this feature compared to those which do not and measured the safety benefit associated with the provision of a shoulder belt in the center rear seat position. The data demonstrate that the presence of a shoulder belt in the center rear seating position influences seating practices only when there is a single child occupant in the vehicle. Belted children in the center rear of vehicles equipped with a lap shoulder belt are at an 81% reduction in risk of injury than those belted in the center rear equipped with a lap only belt. The data suggest that by requiring lap shoulder belts in the center rear, benefits would be realized to belted children, specifically the 4-8 year old group.


[Embolization of iatrogenic renal vascular lesions with coaxial catheters and microcoils].
We report our experience with the superselective embolization of iatrogenic vascular injuries. We used new coaxial catheters and new microcoils to assess their technical features and efficacy. Diagnostic angiography by catheterization of the main renal artery was performed in 7 patients with suspected renal vascular injury to detect injury presence and site. After super-selective catheterization of the feeding arteries with a coaxial catheter (Tracker 18), we performed embolization with platinum microcoils inserted using a coil pusher. The procedure efficacy was assessed at angiography and by comparing serum hemoglobin and hematocrit levels before and after embolization; posttreatment renal function was also studied. Selective embolization permitted complete occlusion of the arteries feeding the vascular injury in all patients, preserving as much parenchyma as possible. Hemoglobin and hematocrit levels were improved or unchanged after the maneuver (average hemoglobin level before embolization: 9.01 mg/dl versus 9.95 mg/dl after embolization; average hematocrit level before embolization: 27.3% versus 30.06% after embolization). We had no difficulty during microcoil insertion and the coaxial catheter was always stable. Superselective embolization of iatrogenic vascular injuries is an effective and definitive treatment. We recommend it in the patients with renal bleeding due to iatrogenic vascular injury after percutaneous diagnostic or therapeutic procedures.


Effects of photodynamic therapy using verteporfin on experimental choroidal neovascularization and normal retina and choroid up to 7 weeks after treatment.
To study the long-term effects of photodynamic therapy (PDT), using liposomal benzoporphyrin derivative (BPD) or Verteporfin, on experimental choroidal neovascularization (CNV) and on normal retina and choroid (with no CNV) in the cynomolgus monkey eye. Photodynamic therapy was performed in 8 cynomolgus monkey eyes with experimental CNV induced by laser injury. The effect of PDT on normal retina and choroid (with no CNV) was studied in 9 monkey eyes. Liposomal BPD was administered intravenously (0.375 mg/kg) either as a bolus, as a slow infusion over 32 minutes, or as a fast infusion over 10 minutes. Photodynamic therapy was performed using light at a wavelength of 689 or 692 nm, with an irradiance of 600 mW/cm2 and fluence of 150 J/cm2. Follow-up studies, including fundus photography and FA, were performed at 24 hours after PDT and then weekly. Indocyanine green and BPD angiography were performed in selected cases. Tissues were examined with light and electron microscopy at the end of follow-up. Twenty-three of the 32 areas of CNV treated with PDT showed absence of angiographic leakage at 24 hours. Twenty-eight areas of CNV were followed for 4 weeks; 22 of 28 showed absence of angiographic leakage at 2 weeks; and 20 of 28 at 4 weeks of follow-up. Forty spots on the normal retina and choroid were treated with PDT and were followed for 4 to 7 weeks. These spots showed pigment-laden cells in the outer retina, variably pigmented retinal pigment epithelium (RPE) in the treated area, intact neurosensory retina, and reperfusion of the choriocapillaris. Photodynamic therapy leads to absence of angiographic leakage for at least 4 weeks in experimental CNV in the monkey model. In the normal monkey eye the RPE and choriocapillaris show generalized recovery with preservation of the neurosensory retina 7 weeks after PDT.


Antibacterial, anthelmintic, and analgesic activities of Piper sylvaticum (Roxb.) leaves and in silico molecular docking and PASS prediction studies of its isolated compounds.
Background In the present study, we investigated the antibacterial, anthelmintic, and analgesic activities of methanol extract of P. sylvaticum leaves (MEPSL) in experimental models. Then, computational analysis (in silico molecular docking and PASS prediction) was performed to determine the potent phytoconstituents of total six isolated compounds of this plant for antibacterial and anthelmintic activities. Methods Qualitative and quantitative phytochemical studies were carried out by established methods. In vitro antibacterial activity was determined by disc diffusion technique and anthelmintic activity was tested against Tubifex tubifex worm whereas analgesic activity was determined by the acetic acid-induced writhing test in mice. Molecular docking study was performed using Schrödinger Maestro 10.1 and an online tool used for PASS prediction. Results Our phytochemical study revealed the presence of alkaloids, flavonoids, saponins, and also indicated a substantial amount of phenols (65.83 mg), flavonoids (102.56 mg), and condensed tannins (89.32 mg). MEPSL showed good antibacterial activity against both gram-positive and gram-negative bacteria. Our result exhibited that MEPSL has strong anthelmintic action compared to standard levamisole. In addition, the extract also showed a dose-dependent and statistically significant analgesic activity at the doses of 200 and 400 mg/kg, body weight. Docking studies showed that piperine and piperlonguminine have the best scores for the tested enzymes. PASS predicted the antibacterial and anthelmintic activity of both phytoconstituents. Conclusions This study suggests that MEPSL possess significant antibacterial, anthelmintic, and analgesic activities which could be related to the presence of several phytochemicals. The phytoconstituents, i.e. piperine and piperlonguminine were found to be most effective in computational studies.


Is baseline medication resistance associated with potential for relapse after successful remission of a depressive episode with ECT? Data from the Consortium for Research on Electroconvulsive Therapy (CORE).
To test whether pre-electroconvulsive therapy (ECT) medication resistance is associated with post-ECT relapse rates. In a post hoc analysis of data from a large multicenter trial of post-ECT relapse prevention strategies (conducted from May 1997 to July 2004), we assessed whether response to antidepressant medications prior to ECT for a unipolar nonpsychotic depressive episode (DSM-IV) was associated with differential relapse rates after remission with ECT. Baseline (i.e., pre-ECT) medication use was assessed with the Antidepressant Treatment History Form. Following remission with ECT that was stable for 1 week, patients were randomly assigned to receive 6 months of treatment with either combination lithium carbonate/nortriptyline or continuation ECT. Relapse was assessed with the 24-item Hamilton Rating Scale for Depression. There were 146 patients followed in the first week after remission (termed the interim week in this study), and 73 in the randomized phase of the study. For the purposes of this trial, medication resistance is defined as not having responded to at least 1 adequate trial of an antidepressant medication before ECT. In the first week after acute remission, 9.8% of patients not having at least 1 antidepressant medication trial met relapse criteria, while 31.4% of medication-resistant patients met relapse criteria, a difference that was statistically significant (p = .026). In the randomized phase of the study, 34.6% of non-medication-resistant patients relapsed, while 50.0% of medication-resistant patients relapsed, a difference that was not significant (p = .434). We conclude that nonpsychotic patients who had at least 1 adequate antidepressant medication trial before ECT may be especially prone to early relapse after successful acute remission with ECT.


Host immune and apoptotic responses to avian influenza virus H9N2 in human tracheobronchial epithelial cells.
The avian influenza virus H9N2 subtype has circulated in wild birds, is prevalent in domestic poultry, and has successfully crossed the species boundary to infect humans. Phylogenetic analyses showed that viruses of this subtype appear to have contributed to the generation of highly pathogenic H5N1 viruses. Little is known about the host responses to H9N2 viruses in human airway respiratory epithelium, the primary portal for viral infection. Using an apically differentiated primary human tracheobronchial epithelial (TBE) culture, we examined host immune responses to infection by an avian H9N2 virus, in comparison with a human H9N2 isolate. We found that IFN-β was the prominent antiviral component, whereas interferon gamma-induced protein 10 kDa (IP-10), chemokine (C-C motif) ligand (CCL)-5 and TNF-α may be critical in proinflammatory responses to H9N2 viruses. In contrast, proinflammatory IL-1β, IL-8, and even IL-6 may only play a minor role in pathogenicity. Apparently Toll-like receptor (TLR)-3, TLR-7, and melanoma differentiation-associated gene 5 (MDA-5) contributed to the innate immunity against the H9N2 viruses, and MDA-5 was important in the induction of IFN-β. We showed that the avian H9N2 virus induced apoptosis through the mitochondria/cytochrome c-mediated intrinsic pathway, in addition to the caspase 8-mediated extrinsic pathway, as evidenced by the cytosolic presence of active caspase 9 and cytochrome c, independent of truncated BH3 interacting domain death agonist (Bid) activation. Further, we demonstrated that FLICE-like inhibitory protein (FLIP), an apoptotic dual regulator, and the p53-dependent Bcl-2 family members, Bax and Bcl-x(s), appeared to be involved in the regulation of extrinsic and intrinsic apoptotic pathways, respectively. The findings in this study will further our understanding of host defense mechanisms and the pathogenesis of H9N2 influenza viruses in human respiratory epithelium.


From concept to reality: the long road to c-Met and RON receptor tyrosine kinase inhibitors for the treatment of cancer.
c-Met and RON are receptor tyrosine kinases (RTK) that are closely related, both from a homology as well as from a functional stand point. Both receptors can induce cell migration, invasion, proliferation and survival in response to their respective ligand. Moreover, both possess oncogenic activity in vitro, in animal models in vivo and are often deregulated in human cancers. c-Met attracted a lot of interest shortly after its discovery in the mid-1980s because of its unusual role in cell motility. Moreover, a causal role for c-Met activating mutations in human cancer propelled an intensive drug discovery effort throughout the research and pharmaceutical communities to find inhibitors of c-Met. While c-Met is now a well-accepted target for an anti-cancer drug, less is known about the role of RON in cancer. Interestingly, despite their many common attributes, c-Met and RON are activated by different mechanisms in cancer cells. Because of the homology between the two RTKs, some small molecule kinase inhibitors of c-Met have inhibitory activity on RON, opening the door to exploring the role of both receptors in human cancers. In this review we will discuss the relevance of both c-Met and RON deregulation in human cancers and the progress so far in identifying small molecule kinase inhibitors that can block the activity of these targets in vitro and lead to anti-tumor effects in animal models.


Characterizing the bioburden of ESBL-producing organisms in a neonatal unit using chromogenic culture media: a feasible and efficient environmental sampling method.
Infections due to extended spectrum beta-lactamase producing organisms (ESBL) have emerged as the leading cause of sepsis among hospitalized neonates in Botswana and much of sub-Saharan Africa and south Asia. Yet, ESBL reservoirs and transmission dynamics within the neonatal intensive care unit (NICU) environment are not well-understood. This study aimed to assess the efficiency and feasibility of a chromogenic-culture-media-based environmental sampling approach to characterize the ESBL bioburden within a NICU. A series of four point-prevalence surveys were conducted at a 36-bed NICU at a public tertiary referral hospital in Botswana from January-June 2021. Samples were collected on 4 occasions under semi-sterile technique using 1) flocked swabs & templates (flat surfaces); 2) sterile syringe & tubing (water aspiration); and 3) structured swabbing techniques (hands & equipment). Swabs were transported in physiological saline-containing tubes, vortexed, and 10 µL was inoculated onto chromogenic-agar that was selective and differential for ESBL (CHROMagar™ ESBL, Paris, France), and streaking plates to isolate individual colonies. Bacterial colonies were quantified and phenotypically characterized using biochemical identification tests. In total, 567 samples were collected, 248 (44%) of which grew ESBL. Dense and consistent ESBL contamination was detected in and around sinks and certain high-touch surfaces, while transient contamination was demonstrated on medical equipment, caregivers/healthcare worker hands, insects, and feeding stations (including formula powder). Results were available within 24-72 h of collection. To collect, plate, and analyse 50 samples, we estimated a total expenditure of $269.40 USD for materials and 13.5 cumulative work hours among all personnel. Using basic environmental sampling and laboratory techniques aided by chromogenic culture media, we identified ESBL reservoirs (sinks) and plausible transmission vehicles (medical equipment, infant formula, hands of caregivers/healthcare workers, & insects) in this NICU environment. This strategy was a simple and cost-efficient method to assess ESBL bioburden and may be feasible for use in other settings to support ongoing infection control assessments and outbreak investigations.


Giant labial fibroepithelial stromal polyp.
We report an18-year-old girl with a four-year history of a slow-growing labial mass with a sudden increase in size in the last year. Examination revealed a large fleshy 20 cm perineal mass centering on the left labia majora and attached to it by a 1cm pedicle. It was associated with pain, ulceration and discharge. The lesion was excised via diathermy at the base of the stalk. The excised specimen weighed 1.112kg and measured 20.5 x 17 x 5cm. The lesion showed a solid, soft whitish, cut surface. Histology revealed a hypocellular tumour with focally oedematous fibrous stroma in which were scattered large and small blood vessels, mast cells and other chronic inflammatory cells. True myxoid matrix was not observed. The stromal cells had a spindle to stellate morphology. There was no significant cytological atypia, mitotic activity or necrosis. The tumour cells were negative for SMA, desmin, CD34, S100 protein, EMA and PR. The diagnosis was clinically and histologically challenging because various vulvovaginal soft tissue tumours often have overlapping clinicopathological features. However, based on strict histological criteria and the absence of worrisome cytological features, a diagnosis of fi broepithelial stromal polyp was rendered despite the unusual size. A review of the literature shows that whilst vulvovaginal fibroepithelial stromal polyps are well described, giant variants are rare. Awareness of the extraordinary size that can be attained by such polyps can facilitate swift clinical and histological diagnosis.


[Periorbital pallor post application of mydriatic in infants with hydrocephalus associated to systemic effects].
Adequate pupil dilation is needed to evaluate some neonates at risk of developing illness during this stage. However, this procedure is not free of adverse effects, either local or systemic. One of these complications is the local vasoconstriction of the preterm baby's skin following the application of mydriatic eye drops. To describe secondary local and systemic complications of pharmacological pupil dilation in 2 newborns. Clinical case 1: Full term baby with diagnosis of low-birth weight and hydrocephalia. An ophtalmological evaluation was performed at 5 days of age due to the presence of corneal opacities. Peri ocular pallor was observed during the procedure, as well as tachycardia and hypertension 2 hours later, spontaneosly recovered. Case 2: Preterm newborn, 27 weeks of gestational age. Neonatal respiratory distress syndrome, patent ductus arteriosus, intraventricular hemorrhage and hydrocephalia were diagnosed at birth. At 28 days of life an ophtalmological evaluation was performed. After 10 minutes of mydriatic drops administration to evaluate preterm retinopathy, peri ocular pallor was observed, with spontaneous resolution; however, 24 hours later, the patient showed abdominal distention and feeding intolerance. Necrotizing enterocolitis was discarded, and symptoms were spontaneosly recovered. The establishment of protocols in relation to the number of drops to apply for dilation is needed to reduce deleterious effects on high risk infants, such as premature babies and those with hydrocephalus. Therefore this monitoring practice should be performed during the evaluation.


Apoptotic and anti-apoptotic synaptic signaling mechanisms.
Although several prominent morphological features of apoptosis are evident in the cell body (e.g., cell shrinkage, membrane blebbing, and nuclear DNA condensation and fragmentation) the biochemical and molecular cascades that constitute the cell death machinery can be engaged in synaptic terminals and neurites. Initiating events such as oxyradical production and calcium influx, and effector processes such as Par-4 production, mitochondrial alterations and caspase activation, can be induced in synapses and neurites. Several prominent signal transduction pathways in synaptic terminals play important roles in either promoting or preventing neuronal death in physiological and pathological conditions. For example, activation of glutamate receptors in postsynaptic spines can induce neuronal apoptosis, whereas local activation of neurotrophic factor receptors in presynaptic terminals can prevent neuronal death. Factors capable of inducing nuclear chromatin condensation and fragmentation can be produced locally in synaptic terminals and neurites, and may propogate to the cell body. Recent findings suggest that, beyond their roles in inducing or preventing cell death, apoptotic and anti-apoptotic cascades play roles in synaptic plasticity (structural remodelling and long-term functional changes). For example, caspase activation results in proteolysis of glutamate receptor (AMPA) subunits, which results in altered neuronal responsivity to glutamate. Activation of neurotrophic factor receptors in synaptic terminals can result in local changes in energy metabolism and calcium homeostasis, and can induce long-term changes in synaptic transmission. The emerging data therefore suggest that synapses can be considered as autonomous compartments in which both pro- and anti-apoptotic signaling pathways are activated resulting in structural and functional changes in neuronal circuits. A better understanding of such synaptic signaling mechanisms may reveal novel approaches for preventing and treating an array of neurodegenerative conditions that are initiated by perturbed synaptic homeostasis.


Linking proactive personality and the Big Five to motivation to learn and development activity.
The authors investigated links between the Big Five, proactive personality, and motivation to learn. Web-based survey data were collected at 2 points in time from 183 employees of a financial services firm. Results showed that proactive personality was, only in part, a composite of Big Five facets, which accounted for 26% of its variance. Structural equation modeling results demonstrated that proactive personality, openness, extraversion, and conscientiousness predicted motivation to learn. In addition, motivation to learn was positively related to objectively assessed development activity. Proactive personality, extraversion, and openness had significant indirect links to development activity. Hierarchical regression results suggested that proactive personality had significant incremental validity in the prediction of motivation to learn over all relevant Big Five facets.


Colonoscopy after curative resection of colorectal cancer.
Colonoscopy is generally considered to be an important part of the follow-up program for patients who have undergone curative resection of colorectal cancer. However, there are few data available concerning the frequency with which colonoscopy should be performed and for what length of time after operation. Since 1978, our policy has been to examine the colon annually in these patients using colonoscopy alternating with barium enema. We have evaluated the results in 100 patients over a four-year period. Based on size and histology, the significant colonoscopic findings were new colon cancers in three patients and 11 polyps demonstrating increased risk for malignancy in nine patients. This represents an interval yield of 3% per year. From these results and other reports, we recommend that these patients undergo total colonoscopy in the perioperative period to identify and remove synchronous lesions of the colon, and that examination of the remaining colon should be performed annually, preferably with colonoscopy, for at least the first four years after curative resection.


In vitro and in vivo imaging with quantum dots.
Quantum dots (QDs), also named semiconductor nanocrystals, have initiated a new realm of bioscience by combining nanomaterials with biology, which will profoundly influence future biological and biomedical research. In this review, we describe the extraordinary optical properties of QDs and developments in methods for their synthesis. We focus on fluorescent imaging with QDs both in vitro and in vivo, and the cytotoxicity of QDs and potential barriers to their use in practical biomedical applications. Finally, we provide insights into improvements aimed at decreasing the cytotoxicity of QDs and the future outlook of QD applications in biomedical fields.


Prevalence and incidence rates of atrial fibrillation in Norway 2004-2014.
To study time trends in incidence of atrial fibrillation (AF) in the entire Norwegian population from 2004 to 2014, by age and sex, and to estimate the prevalence of AF at the end of the study period. A national cohort of patients with AF (≥18 years) was identified from inpatient admissions with AF and deaths with AF as underlying cause (1994-2014), and AF outpatient visits (2008-2014) in the Cardiovascular Disease in Norway (CVDNOR) project. AF admissions or out-of-hospital death from AF, with no AF admission the previous 10 years defined incident AF. Age-standardised incidence rates (IR) and incidence rate ratios (IRR) were calculated. All AF cases identified through inpatient admissions and outpatient visits and alive as of 31 December 2014 defined AF prevalence. We identified 175 979 incident AF cases (30% primary diagnosis, 69% secondary diagnosis, 0.6% out-of-hospital deaths). AF IRs (95% confidence intervals) per 100 000 person years were stable from 2004 (433 (426-440)) to 2014 (440 (433-447)). IRs were stable or declining across strata of sex and age with the exception of an average yearly increase of 2.4% in 18-44 year-olds: IRR 1.024 (1.014-1.034). In 2014, the prevalence of AF in the adult population was 3.4%. We found overall stable IRs of AF for the adult Norwegian population from 2004 to 2014. The prevalence of AF was 3.4% at the end of 2014, which is higher than reported in previous studies. Signs of an increasing incidence of early-onset AF (<45 years) are worrying and need further investigation.


[Bullous pemphigoid with figurate erythema. A case report].
Bullous pemphigoid is an autoimmune dermatosis characterized by deposition of immunoglobulin G (IgG) and/or complement along basement membrane zone. The lesions of bullous pemphigoid are tense blisters on trunk and extremities, oral lesions are unusual. Figurate erythema includes some dermatologic pictures characterized by annular and polycyclic lesions. Figurate erythema can be confused with erythema gyratum repens, dermatosis associated statistically to malignant neoplasias. The objective for reporting this case is to inform the simultaneous association between bullous pemphigoid and figurate erythema in 83 year old man without underlying malignancy.


[Submaxillary streptococcal B cellulitis in young infants].
The authors report 3 cases of young infants with B streptococcal cellulitis revealed by submandibular inflammatory induration. The clinical findings, similar to the cases reported in literature, were characteristic, due to the infants' ages, the aspect and localization of the cellulitis. Bacteriologic diagnosis relies on blood cultures and aspiration of the lesion.


Mandibular asymmetry: a proposal of radiographic analysis with public domain software.
This preliminary study aimed to propose a new analysis of digital panoramic radiographs for a differential diagnosis between functional and morphological mandibular asymmetry in children with and without unilateral posterior crossbite. Analysis is based on linear and angular measurements taken from nine anatomic points, demarcated in sequence directly on digital images. A specific plugin was developed as part of a larger public domain image processing software (ImageJ) to automate and facilitate measurements. Since panoramic radiographs are typically subject to magnification differences between the right and left sides, horizontal linear measurements were adjusted for greater accuracy in both sides by means of a Distortion Factor (DF). In order to provide a preliminary assessment of proposed analysis and the developed plugin, radiographs of ten patients (5 with unilateral posterior crossbite and 5 with normal occlusion) were analyzed. Considerable divergence was found between the right and left sides in the measurements of mandibular length and position of condyles in patients with unilateral posterior crossbite in comparison to individuals with normal occlusion. Although there are more effective and accurate diagnostic methods, panoramic radiography is still widespread, especially in emerging countries. This study presented initial evidence that the proposed analysis can be an important resource for planning early orthodontic intervention and, thus, avoid progression of asymmetries and their consequences.


Preparation of Cell Extracts for Purification of Proteins Expressed in Pichia pastoris.
Recovery of intracellular proteins requires disruption of the host cell before the target protein is extracted and isolated. Disruption methods vary depending on the type of cells, the total volume, and the number of samples being processed. For cells enveloped in cell walls (such as yeast), mild techniques such as hypotonic shock are not sufficient to achieve adequate lysis. More vigorous methods are often required. Although the preferred medium- or large-scale method of breaking yeast cells is mechanical shearing, lysis with the aid of glass beads in a BeadBeater is described here.


Routine surgical telepathology in the Department of Veterans Affairs: experience-related improvements in pathologist performance in 2200 cases.
To determine whether diagnostic concordance, case deferral rate, and/or time required to review slides changed significantly as telepathologists gained additional experience using a hybrid dynamic/store-and-forward (HDSF) telepathology (TP) system on the 2000 cases following an initial 200 consecutive surgical cases, previously reported. Gross surgical pathology specimens were prepared by specially trained personnel in Iron Mountain, Michigan. For TP, glass slides were placed on the stage of a robotic microscope at the Iron Mountain VAMC (remote site); control of the motorized microscope was then transferred to a pathologist located 220 miles away at the Milwaukee, Wisconsin, VAMC (host site). For each case, a telepathologist had the option of either rendering a diagnosis or deferring the case for later analysis by conventional light microscopy (LM). After the slides were read by TP and a surgical pathology report had been generated (for nondeferred cases), the slides were transported to Milwaukee, where they were reexamined by the same pathologist, now using LM. When there was disagreement between the TP and LM diagnosis, a supplemental or revised report was issued, and the referring physician was notified by telephone immediately. All supplemental and revised reports were reviewed by a third pathologist in the group. The slides were then reviewed by the pathology group practice or, when there was no consensus, by the Armed Forces Institute of Pathology to establish a "truth" diagnosis. To determine changes in telepathologist performance with experience after the initial start-up of the service, their performance in handling 10 consecutive sets of 200 surgical pathology cases was analyzed. Concordance rates for clinically significant TP and LM diagnoses were high for all 10 sets, ranging from 99% to 100%. Comparing the first set (Cases 201-400) with the last set (Cases 2001-2200), viewing times per case were reduced from 10.26 min to 3. 58 min. Viewing times per slide were reduced from 3.44 min to 1.13 min per slide, comparing the first and last sets. Case turnaround times (TAT) decreased from 2.46 days to < or =1.5 days. Thes results demonstrate that improvements in TP services occur over time as the result of additional experience using the TP system. The high diagnostic concordance and low rate of case deferral lend additional support to the proposal that a host-site pathologist using HDSF TP can substitute effectively for an on-site pathologist as a service provider.


[Clinical-morphological aspects of suppurative inflammatory diseases of the adnexa uteri].
Draining of pyo-inflammatory foci of the uterine tubes, monitored by transvaginal echography, computer-aided tomography and laparoscopy, or surgery were carried out in 169 patients. Morphologic analysis helped define the characteristic features of the most prevalent clinical forms of pyo-inflammatory conditions of the uterine appendages and thus promoted a more accurate diagnosis of such conditions and their differentiated therapy. Immediate and late results of therapeutic draining of the pyo-inflammatory foci in the uterine tubes, monitored by the afore-said methods, are analyzed.


Controlling the orientation of a cell-synthesized extracellular matrix by using engineered gelatin-based building blocks.
In tissue engineering there is growing interest in fabricating highly engineered platforms designed to instruct cells towards the synthesis of tissues that reproduce their natural counterpart. In this context, a fundamental factor to take into account is the control over the final tissue orientation, especially for what concerns the replication of load-bearing tissues whose functions are strictly related to their microstructural organization. Starting from this point, in this work we have engineered a gelatin-based hydrogel in order to be patterned by 2-photon polymerization (2PP) lithography for the fabrication of instructive free standing building blocks designed to produce anisotropic collagen-based μtissues. Biological results clearly highlighted the strong relationship between μtissue orientation and such topographies, which resulted in a crucial element in the production of highly anisotropic μtissues.


The SV40 core sequence functions as a repressor element in yeast.
Our previous studies showed that the AP-1 recognition element (ARE) present within the SV40 72-base pair (bp) enhancer will activate transcription in yeast when placed upstream of a truncated CYC1 promoter. However, the AP-2/AP-3 recognition element (also known as the core sequence TGTGGAAAG) from the SV40 enhancer was not able to activate CYC1-dependent transcription. In this report, we show that the core sequence, when cloned next to a yeast UAS (upstream activation sequence), can inhibit the transcriptional stimulatory activity of the UAS. We refer to this sequence as the upstream repressor element (URE) in yeast. Repression occurs in an orientation-independent fashion and irrespective of the placement of the URE between the UAS and TATA box or upstream of both of these elements. Furthermore, repression is seen when the URE is separated from the UAS by up to 214 bp. Interestingly, multiple copies of an activator site can overcome this repression. Gel-shift analysis and URE-probed proteins blots indicate the presence of two polypeptide chains capable of binding the URE in yeast. The experimental evidence suggests that either the repression associated with the URE sequence is mediated by a direct, one-to-one interaction between the proteins recognizing the URE and GCRE, or alternatively, that there is a direct interaction between the activator and repressor for a general transcription factor.


Molecular investigation against the resistant mechanism of PncA mutated pyrazinamide resistance and insight into the role of pH environment for pyrazinamide activation.
Pyrazinamide (PZA), a crucial component of anti-TB therapy, is a prodrug. PZA interacts with PncA protein to be converted into its functional form i.e. pyrazinoic acid (POA). It has unique feature to kill dormant tubercle bacilli of acidic environment. Although significance of pH environment in PZA activation has been investigated in several of previous studies, insight into the significant atomistic variations in the interaction pattern of PZA with PncA, at different pH environments, are still required to be explored. On the other hand, continuously emerging PncA mutants, associated with PZA resistance, have also become a serious threat for global TB control program. Therefore, the current study was designed to understand the role of pH environment in the PZA activation and to explore the PZA resistance mechanism in various PncA mutants. The study included various in silico experiments like molecular docking, MD simulation, binding free energy estimation, PCA and FEL. In our study, we have found pH-3 and pH-5 environment as a highly significant environment for PZA activation. It was found that protonation or deprotonation of PZA activation site (PAS) residues, majorly K48, D56, K96 and E107, resulted in rearrangement of the PAS according to the pH conditions. It has also been observed that positioning of PZA binding near to Fe2+ and residues of catalytic triad (i.e. D8, K96 and C138) also play a very crucial role in the activation of PZA. The overall insight from the current study may help to develop new therapeutics against PncA mutated PZA resistance.Communicated by Ramaswamy H. Sarma.


Temporal asymmetries in auditory coding and perception reflect multi-layered nonlinearities.
Sound recognition relies not only on spectral cues, but also on temporal cues, as demonstrated by the profound impact of time reversals on perception of common sounds. To address the coding principles underlying such auditory asymmetries, we recorded a large sample of auditory cortex neurons using two-photon calcium imaging in awake mice, while playing sounds ramping up or down in intensity. We observed clear asymmetries in cortical population responses, including stronger cortical activity for up-ramping sounds, which matches perceptual saliency assessments in mice and previous measures in humans. Analysis of cortical activity patterns revealed that auditory cortex implements a map of spatially clustered neuronal ensembles, detecting specific combinations of spectral and intensity modulation features. Comparing different models, we show that cortical responses result from multi-layered nonlinearities, which, contrary to standard receptive field models of auditory cortex function, build divergent representations of sounds with similar spectral content, but different temporal structure.


Insulin receptor substrate-2 (Irs2) in endothelial cells plays a crucial role in insulin secretion.
Endothelial cells are considered to be essential for normal pancreatic β-cell function. The current study attempted to demonstrate the role of insulin receptor substrate-2 (Irs2) in endothelial cells with regard to insulin secretion. Endothelial cell-specific Irs2 knockout (ETIrs2KO) mice exhibited impaired glucose-induced, arginine-induced, and glucagon-induced insulin secretion and showed glucose intolerance. In batch incubation and perifusion experiments using isolated islets, glucose-induced insulin secretion was not significantly different between the control and the ETIrs2KO mice. In contrast, in perfusion experiments, glucose-induced insulin secretion was significantly impaired in the ETIrs2KO mice. The islet blood flow was significantly impaired in the ETIrs2KO mice. After the treatment of these knockout mice with enalapril maleate, which improved the islet blood flow, glucose-stimulated insulin secretion was almost completely restored to levels equal to those in the control mice. These data suggest that Irs2 deletion in endothelial cells leads to a decreased islet blood flow, which may cause impaired glucose-induced insulin secretion. Thus, Irs2 in endothelial cells may serve as a novel therapeutic target for preventing and ameliorating type 2 diabetes and metabolic syndrome.


Precautions in familial transmissible dementia: including familial Alzheimer's disease.
Recent studies suggest that some cases of familial Alzheimer's disease may be associated with a transmissible dementia. Animal experiments show that presymptomatic carriers of "slow virus" agents can transmit disease. Because of these findings, we have extended the precautions previously delineated to include those at risk of acquiring transmissible dementia, specifically, to the descendants of those affected with familial Alzheimer's disease or familial Creutzfeldt-Jakob's disease. Blood donation from such persons may pose a danger, because transmissible spongioform encephalopathy has been passed from animal to animal by blood serum and by the WBC layer of frozen whole blood.


The validity and reliability of a screening questionnaire for Parkinson's disease in a community.
Parkinson's disease is one of the most common neurodegenerative diseases in the elderly population. In order to estimate the prevalence of Parkinson's disease in the community, the application of a good screening tool is essential. We evaluated the validity and reliability of a Parkinson's disease screening questionnaire and propose an alternative measure to improve its validity for use in community surveys. We designed the study in a three-phase approach consisting of a screening questionnaire, neurologic examination, and confirmatory examination. A repeated survey was administered to patients with disease detected in the community and on 150 subjects. We examined internal consistency using Cronbach's alpha test, test-retest reliability using the kappa statistic, and validity using sensitivity, specificity, and ROC curves. Unadjusted odds ratios were utilized for the estimation of weights for each questionnaire item. The Cronbach's alpha of the questionnaire was 0.708. The kappa statistic for test-retest reliability was good to generally fair in most of the items. When newly proposed weighting scores were used, the optimum cut-off value was 7/8. When cut-off value was 5/6 for surveying prevalence in a community, the sensitivity was 0.98, and the specificity was 0.61, with simultaneous improvement in reliability. We recommend 5/6 as the ideal cut-off value for the survey of PD prevalence in community. This questionnaire designed for the Korean community could help future epidemiologic studies of PD.


Echinococcus infections in Chinese dogs: a comparison of coproantigen kits.
Coproantigen test kits for Echinococcus spp. worms in dogs, designed for commercial use, were obtained from three different Chinese producers, and were compared with a laboratory kit using reagents from New Zealand. None of the three producers would provide details of their test validation. From a known set of dog faeces obtained at necropsy from infected and uninfected dogs, and from faeces collected from dogs necropsied in the field, results differed between the kits. For field material, the Tiankang kit showed the best specificity but lacked sensitivity. The Combined kit showed best sensitivity but lacked specificity. Results for the Haitai kit were intermediate. With samples from experimentally infected dogs, both the Haitai and Combined kits lacked sensitivity. Kits will need to be validated by the user before they can be relied on to predict progress in Echinococcus spp. control in China or in other countries.


Dunce mutants of Drosophila melanogaster: mutants defective in the cyclic AMP phosphodiesterase enzyme system.
The cyclic AMP and cyclic GMP phosphodiesterase activities present in flies of six mutant strains of the dunce gene and in the parent wild-type strains are characterized. All of the mutants exhibit aberrant cyclic AMP metabolism. The mutant strains dunceM14, dunceM11, and dunceML appear to be amorphic, because they completely lack the cAMP-specific phosphodiesterase normally present in adult flies. These strains exhibit extremely high levels of cAMP. The mutant strains dunce1, dunce2, and dunceCK are hypomorphic and exhibit reduced levels of the cAMP-specific phosphodiesterase. These strains exhibit less marked increases in cAMP content compared with the three amorphic strains. The dunce2 strain possesses a residual enzyme activity that exhibits anomalous kinetics compared with those of the normal enzyme. The possibility that the dunce locus is the structural gene for the cAMP-specific phosphodiesterase is discussed.


Inhibitors of platelet function in the prevention of arterial thrombosis.
The use of platelet inhibiting drugs in the prevention of arterial thromboembolic occlusive disease demonstrates that only a few trials have been adequately designed to provide conclusive results. Dipyridamole is effective in preventing the thromboembolic complications of prosthetic heart valves and sulfinpyrazone reduces occlusive events in A-V silastic cannulae, while clofibrate is clearly ineffective in secondary prevention of myocardial infarction. No antiplatelet agent has yet been convincingly shown to exert any antithrombotic effect in venous thromboembolism, although the results with hydroxychloroquine deserve further study. Currently, only two antiplatelet drugs, dipyridamole and sulfinpyrazone are clearly proven clinically effective in reducing arterial thromboembolism. The promising results using ASA are under active study.


Tuning optimum transfection of gemini surfactant-phospholipid-DNA nanoparticles by validated theoretical modeling.
Gemini nanoparticles (NPs) are a family of non-viral gene delivery systems with potential for applications in non-invasive gene therapy. Translation of these non-viral gene delivery systems requires improvement of transfection efficiency (TE) through fine-tuning of their physicochemical properties such as electric charge and exact ratios of their components. Since high-throughput experimental screening of minute differences in NP compositions is not routinely feasible, we have developed a coarse-grained model to quantitatively study the energetics of the formation of gene delivery complexes with cationic gemini surfactants (G) (m-s-m type) and helper lipids (H) (1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) and DOPE/1,2-dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC)), in order to use it as a tool to predict effective compositions. The model is based on the polymorphic structural conformational flip of NPs and incorporates the electrostatic, entropic and elastic energies, to predict the formation energy and stability of different polymorphic structures as a function of the electric charge of cationic surfactants and concentration of constituent helper lipids. Our results show that these two factors are intertwined in determining the behavior of gene delivery vectors. Specifically, we show that increasing H/G lowers free energy per DNA base pair and increases the stability of the complex. At pH 7, low H/G and charge ratio (ρ±), where the lamellar structure is favored, the formation free energy per DNA base pair is between 0 and -14kBT. At higher values of H/G (2-3) and ρ±, where HII and cubic structures are formed, the formation free energy drops down to values ≈-50kBT, indicating the stable existence of these polymorphic structures in the NPs. At pH 5, the structural transformation of NPs in the endosomes to the lamellar/HII structure with free energy values of about -40kBT is beneficial for endosomal escape, and correlates with increased transfection efficiency. The theoretical model is supported by transfection data in A7 astrocytes with a panel of 16-3-16 gemini NPs, which validates the mathematical model and supports the hypothesis that the NP polymorphic phase transition increases transfection efficiency.


Implementing methadone medical maintenance in community-based clinics: disseminating evidence-based treatment.
Methadone medical maintenance (MMM) is an effective intervention that minimizes the demands of opioid agonist treatment without compromising good treatment response. Despite the benefits of MMM to both patients and treatment programs, little information is available to help community-based programs implement MMM and select patients who might benefit from this intervention. This study evaluates the impact of a seven-session seminar presentation combined with optional on-site consultation on subsequent changes in clinical programming and on the opinions of community-based treatment staff (n = 96) in five methadone maintenance treatment clinics regarding both the adoption of an MMM protocol and the use of an adaptive stepped care model to deliver it. The presentations were developed based on results from a randomized clinical trial (King, V. L., Kidorf, M. S., Stoller, K. B., Schwartz, R., Kolodner, K., Brooner, R. K. (2006) A 12-month controlled trial of methadone medical maintenance integrated into an adaptive treatment model. Journal of Substance Abuse Treatment 31, 385-393.) together with other studies of MMM to help program staff (a) understand the risks and benefits of MMM, (b) develop criteria to choose who may benefit from MMM, and (c) implement an adaptive stepped care delivery system that includes MMM as the least restrictive level of care. A survey of clinic staff opinion about MMM and stepped care was administered at baseline and at five other points over the course of the 1-year project. Overall, the presentations were rated highly favorable for content and presentation (3.3 on a 4-point scale). At the 12-month follow-up, staff were more likely to believe that MMM facilitates patient participation in community-based rehabilitation oriented activities (p = .026) and that MMM patients receive adequate counseling services (p = .025) and were more likely to support treatment that matches patients who are stable with minimal intensities of care (p = .041). One clinic modified its routine care to an adaptive stepped care model in response to the presentations, and 3 of the 5 clinics used MMM levels of treatment intensity at the end of the project. The results suggest that seminar presentations combined with on-site consultation may be a beneficial mechanism for helping staff at community-based programs learn about and adopt effective interventions developed and tested using rigorous research designs.


[Morphologic features of the alcoholic encephalopathy of Gaillet-Wernicke and problems concerning its diagnosis at autopsy].
The paper is concerned with a study of 4 cases of Gaillet-Wernicke alcoholic encephalopathy. In all cases there was a profound liver pathology. The changes found in the brain were quite typical for this disease, although they differed in the degree and severity of the vessel damage, glial elements and neurons. In one case there was a "hepatic" glia and in one (developing with hemochromatosis) - a deposition of iron salts, especially in the area of posterior hypothalamus and in the crus cerebri tentorium.


[Pilot study on predictive value of plasmatic levels of 9 angiogenetic biomarkers in selection of patients candidate to prostate biopsy].
To reduce the number of negative prostate biopsies in patients with elevated PSA serum levels represents a major challenge in urological oncology. Angiogenetic factors might be involved in initial stages of prostate cancer and might represent useful tools in patients' selection for prostate biopsy. The plasmatic levels of Angiopoietin-2, Follistatin, G-CSF, HGF, IL-8, Leptin, PDGF-BB, PECAM-1 and VEGF were measured by BioPlex immunoassay in patients undergoing prostate biopsy for palpable prostate nodule and/or elevated PSA levels (≥4 ng/mL). They were related with biopsy results. ROC curve analysis was exploited to test the diagnostic accuracy of each biomarker by AUC calculation. A potential cut-off level was computed. Fifty patients were entered. Median PSA was 6.8 ng/mL. A prostate nodule was palpable in 18 (36%) patients. The median number of biopsy cores was 12. Prostate cancer was detected in 25 (50%) and ASAP and PIN in 2 more patients (4%) respectively. Among the 9 considered biomarkers, only leptin showed an interesting diagnostic performance with an AUC of 0.781, at a cut-off value of 2.11 ng/mL, demonstrating a sensitivity of 78%, a specificity of 77% and a positive predictive value of 85%. Main limitations of our study are the exploratory design and the criteria adopted for patients' selection determining a detection rate for prostate cancer above the usual range. Leptin only, in our preliminary study, shows promising diagnostic accuracy for the selection of patients candidate to prostate biopsy. Further studies are required to confirm its diagnostic value and its relation with BMI.


An Efficient Dynamic Programming Algorithm for Phosphorylation Site Assignment of Large-Scale Mass Spectrometry Data.
Phosphorylation site assignment of large-scale data from high throughput tandem mass spectrometry (LC-MS/MS) data is an important aspect of phosphoproteomics. Correct assignment of phosphorylated residue(s) is important for functional interpretation of the data within a biological context. Common search algorithms (Sequest etc.) for mass spectrometry data are not designed for accurate site assignment; thus, additional algorithms are needed. In this paper, we propose a linear-time and linear-space dynamic programming strategy for phosphorylation site assignment. The algorithm, referred to as PhosSA, optimizes the objective function defined as the summation of peak intensities that are associated with theoretical phosphopeptide fragmentation ions. Quality control is achieved through the use of a post-processing criteria whose value is indicative of the signal-to-noise (S/N) properties and redundancy of the fragmentation spectra. The algorithm is tested using experimentally generated data sets of peptides with known phosphorylation sites while varying the fragmentation strategy (CID or HCD) and molar amounts of the peptides. The algorithm is also compatible with various peptide labeling strategies including SILAC and iTRAQ. PhosSA is shown to achieve > 99% accuracy with a high degree of sensitivity. The algorithm is extremely fast and scalable (able to process up to 0.5 million peptides in an hour). The implemented algorithm is freely available at http://helixweb.nih.gov/ESBL/PhosSA/ for academic purposes.


Superresolution modeling using an omnidirectional image sensor.
Recently, many virtual reality and robotics applications have been called on to create virtual environments from real scenes. A catadioptric omnidirectional image sensor composed of a convex mirror can simultaneously observe a 360-degree field of view making it useful for modeling man-made environments such as rooms, corridors, and buildings, because any landmarks around the sensor can be taken in and tracked in its large field of view. However, the angular resolution of the omnidirectional image is low because of the large field of view captured. Hence, the resolution of surface texture patterns on the three-dimensional (3-D) scene model generated is not sufficient for monitoring details. To overcome this, we propose a high resolution scene texture generation method that combines an omnidirectional image sequence using image mosaic and superresolution techniques.


A circuitry and biochemical basis for tuberous sclerosis symptoms: from epilepsy to neurocognitive deficits.
Tuberous sclerosis complex (TSC) is an autosomal dominant monogenetic disorder that is characterized by the formation of benign tumors in several organs as well as brain malformations and neuronal defects. TSC is caused by inactivating mutations in one of two genes, TSC1 and TSC2, resulting in increased activity of the mammalian Target of Rapamycin (mTOR). Here, we explore the cytoarchitectural and functional CNS aberrations that may account for the neurological presentations of TSC, notably seizures, hydrocephalus, and cognitive and psychological impairments. In particular, recent mouse models of brain lesions are presented with an emphasis on using electroporation to allow the generation of discrete lesions resulting from loss of heterozygosity during perinatal development. Cortical lesions are thought to contribute to epileptogenesis and worsening of cognitive defects. However, it has recently been suggested that being born with a mutant allele without loss of heterozygosity and associated cortical lesions is sufficient to generate cognitive and neuropsychiatric problems. We will thus discuss the function of mTOR hyperactivity on neuronal circuit formation and the potential consequences of being born heterozygous on neuronal function and the biochemistry of synaptic plasticity, the cellular substrate of learning and memory. Ultimately, a major goal of TSC research is to identify the cellular and molecular mechanisms downstream of mTOR underlying the neurological manifestations observed in TSC patients and identify novel therapeutic targets to prevent the formation of brain lesions and restore neuronal function.


The FDA extended warning for intravenous haloperidol and torsades de pointes: how should institutions respond?
In September 2007, the Food and Drug Administration (FDA) strengthened label warnings for intravenous (IV) haloperidol regarding QT prolongation (QTP) and torsades de pointes (TdP) in response to adverse event reports. Considering the widespread use of IV haloperidol in the management of acute delirium, the specific FDA recommendation of continuous electrocardiogram (ECG) monitoring in this setting has been associated with some controversy. We reviewed the evidence for the FDA warning and provide a potential medical center response to this warning. Cases of intravenous haloperidol-related QTP/TdP were identified by searching PubMed, EMBASE, and Scopus databases (January 1823 to April 2009) and all FDA MedWatch reports of haloperidol-associated adverse events (November 1997 to April 2008). A total of 70 of IV haloperidol-associated QTP and/or TdP were identified. There were 54 reports of TdP; 42 of these events were reportedly preceded by QTP. When post-event QTc data were reported, QTc was prolonged >450 msec in 96% of cases. Three patients experienced sudden cardiac arrest. Sixty-eight patients (97%) had additional risk factors for TdP/prolonged QT, most commonly receipt of concomitant proarrhythmic agents. Patients experiencing TdP received a cumulative dose of 5 mg to 645 mg, patients with QTP alone received a cumulative dose of 2 mg to 1540 mg. While administration of IV haloperidol can be associated with QTP/TdP, this complication most often took place in the setting of concomitant risk factors. Importantly, the available data suggest that a total cumulative dose of IV haloperidol of <2 mg can safely be administered without ongoing electrocardiographic monitoring in patients without concomitant risk factors.


Terahertz spectroscopy of anisotropic materials using beams with rotatable polarization.
We introduce a polarization-resolved terahertz time-domain spectrometer with a broadband (0.3-2.5 THz), rotatable THz polarization state, and which exhibits minimal change in the electric field amplitude and polarization state upon rotation. This was achieved by rotating an interdigitated photoconductive emitter, and by detecting the orthogonal components of the generated THz pulse via electro-optic sampling. The high precision (<0.1°) and accuracy (<1.0°) of this approach is beneficial for the study of anisotropic materials without rotating the sample, which can be impractical, for instance for samples held in a cryostat. The versatility of this method was demonstrated by studying the anisotropic THz optical properties of uniaxial and biaxial oxide crystals. For uniaxial ZnO and LaAlO3, which have minimal THz absorption across the measurement bandwidth, the orientations of the eigenmodes of propagation were conveniently identified as the orientation angles that produced a transmitted THz pulse with zero ellipticity, and the birefringence was quantified. In CuO, a multiferroic with improper ferroelectricity, the anisotropic THz absorption created by an electromagnon was investigated, mapping its selection rule precisely. For this biaxial crystal, which has phonon and electromagnon absorption, the polarization eigenvectors exhibited chromatic dispersion, as a result of the monoclinic crystal structure and the frequency-dependent complex refractive index.


Prolonged hypothalamic-pituitary-adrenal axis activation after acute coronary syndrome in the GENESIS-PRAXY cohort.
Background Glucocorticoid excess has been linked with cardiovascular disease. Little is known about the long-term cortisol response in patients after acute coronary syndrome. Design The objective of this study was to describe the distribution of salivary cortisol in the post-acute phase of acute coronary syndrome and to describe the association of late-night salivary cortisol with cardiovascular risk factors. Methods We used late-night salivary cortisol measurements post-discharge to estimate hypothalamic-pituitary-adrenal axis activity in 309 patients aged 18-55 years enrolled in the GENESIS-PRAXY study from January 2009-April 2013. We evaluated hypothalamic-pituitary-adrenal axis activity and its association with hypertension, dyslipidemia, diabetes, smoking, family history, prior acute coronary syndrome, psychiatric diseases, acute coronary syndrome severity, as well as mortality and rate of rehospitalization at 12 months. Results Persistently elevated late-night salivary cortisol>2.92 nmol/l was seen in 99 (32.0%) patients: within the range of what may be seen in Cushing's disease. Elevated late-night salivary cortisol was associated with previous acute coronary syndrome (13.3% vs 24.2%, p = 0.02), peripheral vascular disease (3.8% vs 13.1%, p = 0.002), and smoking (32.9% vs 46.5% p = 0.02). Elevated late-night salivary cortisol was associated with higher hemoglobin A1c values (5.6 ± 3.0 vs 6.1 ± 2.9, p = 0.008) and lower high density lipoprotein values (0.94 ± 0.53 vs 0.86 ± 0.50, p = 0.01). There were no differences in psychiatric symptom scores, acute coronary syndrome severity or mortality, and rate of rehospitalization at 12 months. Conclusions Many patients post-acute coronary syndrome have prolonged, marked activation of the hypothalamic-pituitary-adrenal axis. Late-night salivary cortisol co-associates with several cardiovascular risk factors. Further studies are needed to confirm the exact role of hypothalamic-pituitary-adrenal axis activity in the pathophysiology of cardiovascular disease.


Radiotherapy practices in Sweden compared to the scientific evidence.
A systematic assessment of radiotherapy for cancer was conducted by The Swedish Council on Technology Assessment in Health Care (SBU) in 2001. The assessment included a critical review of the literature on radiotherapy for cancer published in 1994-2001 and a prospective survey of radiotherapy practice in Sweden during 12 weeks in the autumn of 2001. The results of the survey were compared with the evidence derived from the scientific literature, and the following conclusions could be drawn: Radiotherapy is currently given to approximately 47% of new cancer cases This figure is on a par with rates reported from other countries. For most tumour types, curative radiotherapy practices in Sweden are generally supported by the literature. Interstitial brachytherapy has been included in the treatment of non-gynaecological malignancies, and prostate cancer is now the most common indication. Palliative radiotherapy has increased and is today given in a more rational way using single or few fractions However, it still seems to be under-utilized in Sweden. The need for radiotherapy can be expected to increase until the year 2010.


Incidence and Medicolegal Significance of Wormian Bones in Human Skulls in North India Region.
Wormian bones (Wbs) are small bones located in or near the sutures of the skull which are irregular in size, shape, and number. The present study is to look into the morphological details of such supernumerary bones of skull with regards to their incidence, number, and topography in skulls from the Haryana region. This observational study was carried out in the tertiary care hospital, in the region of Haryana and the total of 147 human skulls were examined during routine autopsy procedures with respect to sutures and topographic distribution of Wbs. The data were statistically analyzed. In the present study, the overall incidence of Wbs was seen in 52 skulls (35.3%) with males having 23.8% and females having 11.5%. The knowledge of incidence and distribution of Wbs in the skull could be useful to radiologists, causality medical officers, and autopsy surgeons in successfully differentiating a skull fracture from normal suture with Wbs or exit gunshot wound from fractured small Wb and thus ruling out the possibilities of presence or absence of fractures due to physical abuse, brittle bones, and exit gunshot wounds. It is also helpful to neurosurgeons in timely diagnosis and management of diseases or fractures in relation to the Wbs.


Bioinformatics and medical informatics: collaborations on the road to genomic medicine?
In this report, the authors compare and contrast medical informatics (MI) and bioinformatics (BI) and provide a viewpoint on their complementarities and potential for collaboration in various subfields. The authors compare MI and BI along several dimensions, including: (1) historical development of the disciplines, (2) their scientific foundations, (3) data quality and analysis, (4) integration of knowledge and databases, (5) informatics tools to support practice, (6) informatics methods to support research (signal processing, imaging and vision, and computational modeling, (7) professional and patient continuing education, and (8) education and training. It is pointed out that, while the two disciplines differ in their histories, scientific foundations, and methodologic approaches to research in various areas, they nevertheless share methods and tools, which provides a basis for exchange of experience in their different applications. MI expertise in developing health care applications and the strength of BI in biological "discovery science" complement each other well. The new field of biomedical informatics (BMI) holds great promise for developing informatics methods that will be crucial in the development of genomic medicine. The future of BMI will be influenced strongly by whether significant advances in clinical practice and biomedical research come about from separate efforts in MI and BI, or from emerging, hybrid informatics subdisciplines at their interface.


Performance of high-throughput DNA quantification methods.
The accuracy and precision of estimates of DNA concentration are critical factors for efficient use of DNA samples in high-throughput genotype and sequence analyses. We evaluated the performance of spectrophotometric (OD) DNA quantification, and compared it to two fluorometric quantification methods, the PicoGreen assay (PG), and a novel real-time quantitative genomic PCR assay (QG) specific to a region at the human BRCA1 locus. Twenty-Two lymphoblastoid cell line DNA samples with an initial concentration of approximately 350 ng/uL were diluted to 20 ng/uL. DNA concentration was estimated by OD and further diluted to 5 ng/uL. The concentrations of multiple aliquots of the final dilution were measured by the OD, QG and PG methods. The effects of manual and robotic laboratory sample handling procedures on the estimates of DNA concentration were assessed using variance components analyses. The OD method was the DNA quantification method most concordant with the reference sample among the three methods evaluated. A large fraction of the total variance for all three methods (36.0-95.7%) was explained by sample-to-sample variation, whereas the amount of variance attributable to sample handling was small (0.8-17.5%). Residual error (3.2-59.4%), corresponding to un-modelled factors, contributed a greater extent to the total variation than the sample handling procedures. The application of a specific DNA quantification method to a particular molecular genetic laboratory protocol must take into account the accuracy and precision of the specific method, as well as the requirements of the experimental workflow with respect to sample volumes and throughput. While OD was the most concordant and precise DNA quantification method in this study, the information provided by the quantitative PCR assay regarding the suitability of DNA samples for PCR may be an essential factor for some protocols, despite the decreased concordance and precision of this method.


Astrocytic gap junctional communication decreases neuronal vulnerability to oxidative stress-induced disruption of Ca2+ homeostasis and cell death.
We investigated the effect of uncoupling astrocytic gap junctions on neuronal vulnerability to oxidative injury in embryonic rat hippocampal cell cultures. Mixed cultures (neurons growing on an astrocyte monolayer) treated with 18-alpha-glycyrrhetinic acid (GA), an uncoupler of gap junctions, showed markedly enhanced generation of intracellular peroxides (2,7-dichlorofluorescein fluorescence), impairment of mitochondrial function [(dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reduction], and cell death (lactate dehydrogenase release) following exposure to oxidative insults (FeSO4 and 4-hydroxynonenal). GA alone had little or no effect on basal levels of peroxides, mitochondrial function, or neuronal survival. Intercellular dye transfer analyses revealed extensive astrocyte-astrocyte coupling but no astrocyte-neuron or neuron-neuron coupling in the mixed cultures. Studies of pure astrocyte cultures and microscope analyses of neurons in mixed cultures showed that the increased oxidative stress and cell death in GA-treated cultures occurred only in neurons and not in astrocytes. Antioxidants (propyl gallate and glutathione) blocked the death of neurons exposed to FeSO4/GA. Elevations of neuronal intracellular calcium levels ([Ca2+]i) induced by FeSO4 were enhanced in neurons in mixed cultures exposed to GA. Removal of extracellular Ca2+ and the L-type Ca2+ channel blocker nimodipine prevented impairment of mitochondrial function and cell death induced by FeSO4 and GA, whereas glutamate receptor antagonists were ineffective. Finally, GA exacerbated kainate- and FeSO4-induced injury to pyramidal neurons in organotypic hippocampal slice cultures. The data suggest that interastrocytic gap junctional communication decreases neuronal vulnerability to oxidative injury by a mechanism involving stabilization of cellular calcium homeostasis and dissipation of oxidative stress.


Relating protein adduction to gene expression changes: a systems approach.
Modification of proteins by reactive electrophiles such as the 4-hydroxy-2-nonenal (HNE) plays a critical role in oxidant-associated human diseases. However, little is known about protein adduction and the mechanism by which protein damage elicits adaptive effects and toxicity. We developed a systems approach for relating protein adduction to gene expression changes through the integration of protein adduction, gene expression, protein-DNA interaction, and protein-protein interaction data. Using a random walk strategy, we expanded a list of responsive transcription factors inferred from gene expression studies to upstream signaling networks, which in turn allowed overlaying protein adduction data on the network for the prediction of stress sensors and their associated regulatory mechanisms. We demonstrated the general applicability of transcription factor-based signaling network inference using 103 known pathways. Applying our workflow on gene expression and protein adduction data from HNE-treatment not only rediscovered known mechanisms of electrophile stress but also generated novel hypotheses regarding protein damage sensors. Although developed for analyzing protein adduction data, the framework can be easily adapted for phosphoproteomics and other types of protein modification data.


Evaluation of Vestibular Functions in a Case of Vestibular Migraine With Successful Treatment With Erenumab.
This is the first report of vestibular examinations before and after the successful treatment of vestibular migraine (VM), a common cause of recurrent vertigo, with calcitonin gene-related peptide (CGRP) receptor inhibitor. We evaluated a 42-year-old female with VM and concomitant probable Meniere's disease, whose headache and dizziness have improved promptly with the administration of erenumab, a CGRP receptor inhibitor. The sensorineural hearing loss in pure-tone audiometry, dysfunctions shown in vestibular examinations (cervical and ocular vestibular evoked myogenic potentials), and mild endolymphatic hydrops shown in gadolinium-enhanced inner ear magnetic resonance imaging, all in the right ear, revealed no change compared with those observed before treatment. This case suggests that VM may be treated by blocking CGRP in the trigeminal ganglion, which suppresses the effects on the vestibular nucleus; herein, no effects were observed in the inner ear despite the clear amelioration of dizziness.


"Be ready against cancer, now": direct-to-consumer advertising for genetic testing.
A recent addition to the debate about the benefits and harms of direct-to-consumer (DTC) advertising of medicines and pharmaceuticals is a growing critique of DTC marketing and sale of genetic tests. Academic and policy literatures exploring this issue have, however, tended to focus on the sale of genetic tests, paying rather less attention to the particular implications of advertising. The globalization of broadcast media and ever increasing access to the Internet mean that public exposure to advertising for medical technologies is a reality that national regulatory bodies will be hard pressed to constrain. Working through a case study detailing Myriad Genetics' 2002 pilot advertising campaign for their BRACAnalysis genetic susceptibility test for hereditary breast and ovarian cancer, this paper highlights some of the diverse and often overlooked and unregulated approaches to DTC advertising, and the associated social, ethical and policy implications.


Inhibition of endothelium-derived relaxing factor enhances myocardial stunning in conscious dogs.
Impaired endothelial-dependent vascular responses after coronary artery occlusion (CAO) and reperfusion (CAR) have been investigated extensively. However, it is not known whether impaired endogenous endothelium-derived relaxing factor production affects postischemic myocardial dysfunction, ie, myocardial stunning. Eight dogs were instrumented with an intracoronary catheter and an hydraulic occluder on the left circumflex coronary artery. The effects of a 10-minute CAO randomized with and without intracoronary administration of NG-nitro-L-arginine (L-NA), a nitric oxide (NO) synthesis inhibitor, were compared in the same conscious dogs. Postischemic regional contractile dysfunction in subendocardial and subepicardial as well as transmural wall thickening was measured with ultrasonic dimension crystals, and myocardial blood flow was measured with radioactive microspheres. Intracoronary infusion of L-NA did not affect systemic hemodynamics, and transmural myocardial blood flow was reduced slightly (-8%), but significantly, only in the left circumflex territory. The recovery of wall thickening was significantly delayed in the presence of L-NA compared with the absence of L-NA, eg, at 30-minute CAR, not only in the subendocardium (-76 +/- 9% versus -49 +/- 9%) but also in the subepicardium (-52 +/- 8% versus -29 +/- 7%). During CAO, blood flow was decreased identically in both conditions, and during CAR, the differences in blood flow were minor (7%). Inhibition of NO synthesis enhanced myocardial stunning transmurally in conscious dogs, potentially independent of its effects on blood flow.


Interactions between integrin receptors and fibronectin are required for calvarial osteoblast differentiation in vitro.
We previously showed that anti-fibronectin antibodies or soluble fibronectin fragments containing the central cell-binding domain inhibit formation of mineralized nodules by fetal calvarial osteoblasts in vitro. These findings suggest a critical role for fibronectin in osteoblast differentiation and morphogenesis. In this study we tested the hypothesis that fibronectin's effects on osteogenesis are mediated via direct interactions with integrin receptors for fibronectin on osteoblasts. Immunocytochemical analysis identified the integrin fibronectin receptor alpha5ss1 in fetal rat calvarial tissue and in cultured osteoblasts at all stages of differentiation. Three other integrins, alpha3ss1, alpha8ss1 and alphavss3, which can bind fibronectin, as well as other matrix components, were also identified in tissue and at all stages of cell culture. Immunoprecipitation data showed that alpha5ss1 levels are constant throughout osteoblast differentiation whereas levels of alpha3ss1 and alpha8ss1 decline in mature mineralized cultures. To determine whether integrin fibronectin receptors are required for osteoblast formation of mineralized nodules, we examined the extent of nodule formation in the presence and absence of function-perturbing anti-integrin antibodies. The antibodies were present continuously in cultures beginning at confluence (day 3), and nodule formation was measured at days 10 and 20. An anti-alpha5 integrin subunit antibody reduced nodule formation to less than 5% of control values at both time points. Inhibition of nodule formation was reversible and did not affect cell attachment and viability. Function-perturbing antibodies against alpha3ss1 and alpha8ss1 also reduced nodule formation, to less than 20% of control values. In contrast, function-perturbing antibodies to alphavss3 and alphavss5 did not affect nodule formation, indicating that the inhibitions noted were indeed specific. To determine the effect of antibody treatment on gene expression, steady-state mRNA expression was examined and found to be suppressed for osteoblast markers alkaline phosphatase and osteocalcin. Together, these results indicate that direct osteoblast interactions with the extracellular matrix are mediated by a select group of integrin receptors that includes alpha5ss1, alpha3ss1 and alpha8ss1. We further conclude that the specific alpha5ss1 fibronectin receptor mediates critical interactions between osteoblasts and fibronectin required for both bone morphogenesis and osteoblast differentiation.