Protection of rats by clofibrate against the hypoglycaemic and toxic effects of hypoglycin and pent-4-enoate. An ultrastructural and biochemical study.
An ultrastructural and biochemical study of the toxic and hypoglycaemic effects of hypoglycin and pent-4-enoate was made on the livers of normal and clofibrate-fed rats. Injection of hypoglycin to rats doubles (from 22% to 44%) the volume fraction of mitochondria and decreases (from 1.05% to 0.26%) the volume fraction of peroxisomes in hepatocytes. The fast-acting toxin pent-4-enoate causes few ultrastructural changes except for the accumulation of lipids. In male adult rats fed with 0.5% clofibrate in their diet for 1-2 months, the volume fraction occupied by peroxisomes and mitochondria in hepatocytes rose to 6.26% and 29.5% respectively. Clofibrate feeding apparently protected the animals against the toxic, hypoglycaemic and hypothermic effects of hypoglycin and of pent-4-enoate, and completely prevented the ultrastructural damage caused by hypoglycin. After hypoglycin administration, hepatic mitochondrial butyryl-CoA dehydrogenase activity was inhibited by more than 90% and, surprisingly, the activity of the peroxisomal enzymes studied was largely preserved. When hypoglycin was given to rats fed on a clofibrate-containing diet, the oxidation of decanoylcarnitine, which was incomplete after hypoglycin treatment alone, remained incomplete with uncoupled mitochondria, but became apparently complete with coupled mitochondria. In the latter condition, there was a slowing of the rate during the last quarter of the pulse of oxygen uptake. Further, butyryl-CoA dehydrogenase activity was much less affected by hypoglycin in clofibrate-fed animals. Pent-4-enoate does not inhibit beta-oxidation in coupled mitochondria from clofibrate-treated rats.


Increases in central blood volume modulate carotid baroreflex resetting during dynamic exercise in humans.
We sought to determine if resetting of the carotid-vasomotor baroreflex function curve during exercise is modulated by changes in central blood volume (CBV). CBV was increased during exercise by altering: (1) subject posture (supine versus upright) and (2) pedal frequency (80 versus 60 revolutions min(-1) (r.p.m.)); while oxygen uptake ( ) was kept constant. Eight male subjects performed three exercise trials: upright cycling at 60 r.p.m. (control); supine cycling at 60 r.p.m. (SupEX) and upright cycling at 80 r.p.m. to enhance the muscle pump (80EX). During each condition, carotid baroreflex (CBR) function was determined using the rapid neck pressure (NP) and neck suction (NS) protocol. Although mean arterial pressure (MAP) was significantly elevated from rest (88 +/- 2 mmHg) during all exercise conditions (P < 0.001), the increase in MAP was lower during SupEX (94 +/- 2 mmHg) and 80EX (95 +/- 2 mmHg) compared with control (105 +/- 2 mmHg, P < 0.05). Importantly, the blood pressure responses to NP and NS were maintained around these changed operating points of MAP. However, in comparison to control, the carotid-vasomotor baroreflex function curve was relocated downward and leftward when CBV was increased during SupEX and 80EX. These alterations in CBR resetting occurred without any differences in or heart rate between the exercise conditions. Thus, increasing CBV and loading the cardiopulmonary baroreflex reduces the magnitude of exercise-induced increases in MAP and CBR resetting. These findings suggest that changes in cardiopulmonary baroreceptor load influence carotid baroreflex resetting during dynamic exercise.


Efficacy of Glucocorticoids and Calcineurin Inhibitors for Anti-aminoacyl-tRNA Synthetase Antibody-positive Polymyositis/dermatomyositis-associated Interstitial Lung Disease: A Propensity Score-matched Analysis.
The optimal treatment strategy for anti-aminoacyl-tRNA synthetase antibody-positive polymyositis/dermatomyositis-associated interstitial lung disease (anti-ARS-PM/DM-ILD) is yet to be established. We aimed to evaluate the efficacy of glucocorticoids and calcineurin inhibitors (CNI) in patients with ARS-PM/DM-ILD. Progression-free survival (PFS) and overall survival rates were retrospectively evaluated in 32 consecutive patients with ARS-PM/DM-ILD. Disease progression was defined as deterioration in PM/DM-ILD (including recurrence). Predictive factors associated with PFS were analyzed by Cox hazards analysis. The efficacy of first-line prednisolone (PSL) plus CNI therapy was compared with that of PSL monotherapy using propensity score-matched analysis. Overall, 20 (62.5%) and 12 (37.5%) patients received first-line therapy with PSL + CNI and PSL, respectively. The 2-year PFS and 5-year survival rates in the overall cohort were 68.8% and 96.9%, respectively. On multivariate analysis, arterial oxygen pressure (HR 0.86) and PSL monotherapy (vs PSL + CNI; HR 7.29) showed an independent association with PFS. Baseline characteristics of propensity score-matched PSL + CNI and PSL groups were similar. The 2-year PFS rate was significantly higher in the matched PSL + CNI group than in the matched PSL group. All patients who experienced disease progression during first-line therapy were subsequently treated with second-line therapies. The 5-year survival rates of both the matched PSL + CNI and PSL groups were favorable. Propensity score-matched analysis demonstrated that first-line PSL + CNI therapy for patients with ARS-PM/DM-ILD significantly improved the PFS compared with PSL monotherapy, although there was no significant difference regarding longterm survival.


GABA inactivation of visual area MT modifies the responsiveness and direction selectivity of V2 neurons in Cebus monkeys.
We investigated the contribution of the projections from area MT to the receptive field properties of cells in visual area V2 in anesthetized and paralyzed Cebus apella monkeys. We recorded extracellular single-unit activity using tungsten microelectrodes in three monkeys before and after pressure injection of a 0.25-mol/l GABA solution. The visual stimulus consisted of a single bar moving in one of eight directions. In total, 72 V2 neurons were studied in 18 sessions of GABA injection into area MT. A group of 22 neurons was investigated over a shorter period of time ranging from 15 to 60 min, during which the activity did not return to baseline levels. The remaining 50 neurons were studied over a period of at least 2 h, and no statistical difference was observed in the neuronal response before and long after GABA inactivation. The effects on these 50 neurons consisted of an early (1-20 min) significant general decrease in excitability with changes in either orientation or direction selectivity. The differential decrease in excitability resulted in an intermediate improvement (20-40 min) of the signal-to-noise ratio for the stimulus-driven activity. The inactivation depended on the quantity of GABA injected into area MT and persisted for a period of 2 h. The GABA inactivation in area MT produced inhibition of most cells (72%) and a significant change of direction tuning in the majority (56%) of V2 neurons. Both increases and also decreases in the direction tuning of V2 neurons were observed. These feedback projections are capable of modulating not only the levels of spontaneous and driven activity of V2 neurons but also the V2 receptive field properties, such as direction selectivity.


Evidence that clearance of hepatitis C virus RNA after alpha-interferon therapy in dialysis patients is sustained after renal transplantation.
To date, there is no available treatment of hepatitis C virus (HCV) infection after renal transplantation (RT). Among 55 anti-HCV-positive/HCV RNA-positive hemodialysis patients who were treated with IFN-alpha (9 MU/wk during 6 or 12 mo), 21 of them (38%) had a sustained virologic response. Of these, 16 (76%) underwent RT 38 mo (range, 2 to 57 mo) after alpha-IFN therapy. There were 13 men and 3 women aged 46 yr (range, 27 to 68 yr). At RT, HCV serology was still positive in 15 patients, and HCV viremia was negative in all patients. Immunosuppression relied on anticalcineurin agents with or without steroids and/or antimetabolites; in addition, 12 of them received induction therapy with antithymocyte globulins. At the last follow-up after RT, at 22.5 mo (range, 2 to 88 mo), HCV viremia remained negative in all patients. Moreover, HCV RNA was not present in peripheral blood mononuclear cells when assessed in eight patients. HCV serology was found to be still positive in 13 patients. Three patients presented with acute rejection, one presented with a suppurative lymphocele, one died from a sepsis, and four presented with a cytomegalovirus infection. None of them developed posttransplant diabetes mellitus. In conclusion, hemodialysis patients waiting for a RT need to be treated with alpha-IFN because when HCV RNA clearance occurred, they experienced no relapse after transplantation despite chronic immunosuppressive treatment.


Terson's syndrome.
Terson's syndrome is intraocular hemorrhage (IOH) subsequent to subarachnoid hemorrhage (SAH). Its presence is associated with higher mortality in SAH. We report a case of Terson's syndrome and review the literature. A 71-year-old Caucasian gentleman collapsed and became comatose. Past medical history was notable for chronic anticoagulation for previous transient ischemia attacks. CT head scans showed severe SAH of Fisher grade 4 and a lesion suspicious for aneurysm. Formal angiography confirmed a supraclinoid right internal carotid artery aneurysm which was coiled. ICU admission was complicated by a stormy course. The patient eventually regained consciousness and was transferred to a regular ward. On hospital day 20, impaired vision was noted. Review of CT head scans revealed previously missed retinal hemorrhages and funduscopy confirmed vitreous hemorrhage. However, the patient remained in a poor neurologic state and expired several days later. Terson's syndrome occurs in up to 40% of acute aneurysmal bleeds. The sudden spike in intracranial pressure (ICP) with aneurysmal rupture is thought to underlie the cause of IOH as well as the high incidence of coma, higher Hunt and Hess grades, and mortality in these patients. Gold-standard diagnosis is funduscopy, and retinal hemorrhages may occasionally be seen on CT. Terson's syndrome occurs frequently following SAH, although it is under-reported. Suspected visual loss following SAH should prompt a search for Terson's syndrome by funduscopy, as its presence is an adverse prognostic factor.


Regeneration of subcutaneous tissue-engineered mandibular condyle in nude mice.
To explore the feasibility of regenerating mandibular condyles based on cartilage cell sheet with cell bone-phase scaffold compared with cell-biphasic scaffolds. Tissue-engineered mandibular condyles were regenerated by the following: 1) cartilage cell sheet + bone-phase scaffold (PCL/HA) seeded with bone marrow stem cells (BMSCs) from minipigs (cell sheet group), and 2) cartilage phase scaffold (PGA/PLA) seeded with auricular chondrocytes + bone-phase scaffold seeded with BMSCs from minipigs (biphasic scaffold group). They were implanted subcutaneously in nude mice after being cultured in vitro for different periods of time. After 12 weeks, the mice were sacrificed, and the specimens were harvested and evaluated based on gross appearance and histopathologic observations with hematoxylin and eosin, safranin O-fast green and immumohistochemical staining for collagen I and II. The histopathologic assessment score of condylar cartilage and bone density were compared between the 2 groups using SPSS 17.0 software. The 2 groups' specimens all formed mature cartilage-like tissues with numerous chondrocytes, typical cartilage lacuna and abundant cartilage-specific extracellular matrix. The regenerated cartilage was instant, continuous, homogeneous and avascular. In the biphasic scaffold group, there were still a few residual PGA fibers in the cartilage layer. The cartilage and bone interface was established in the 2 groups, and the microchannels of the bone-phase scaffolds were filled with bone tissue. The score of cartilage regeneration in the cell sheet group was a little higher than that in the biphasic scaffold group, but the difference was not significant (p > 0.05). There was no significant difference in bone tissue formation between the 2 groups (p > 0.05). Both the cartilage cell sheet group and the biphasic scaffold group of nude mice underwent regeneration of condyle-shaped osteochondral composite. Without residual PGA fibers, the cell sheet group might have less chance of immunological rejection compared to biphasic scaffold group.


Single dose oral ibuprofen plus caffeine for acute postoperative pain in adults.
There is good evidence that combining two different analgesics in fixed doses in a single tablet can provide better pain relief in acute pain and headache than either drug alone, and that the drug-specific benefits are essentially additive. This appears to be broadly true in postoperative pain and migraine headache across a range of different drug combinations, and when tested in the same and different trials. Adding caffeine to analgesics also increases the number of people obtaining good pain relief. Combinations of ibuprofen and caffeine are available without prescription in some parts of the world. To assess the analgesic efficacy and adverse effects of a single oral dose of ibuprofen plus caffeine for moderate to severe postoperative pain, using methods that permit comparison with other analgesics evaluated in standardised trials using almost identical methods and outcomes. We searched the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, EMBASE, the Oxford Pain Relief Database, two clinical trial registries, and the reference lists of articles. The date of the most recent search was 1 February 2015. Randomised, double-blind, placebo- or active-controlled clinical trials of single dose oral ibuprofen plus caffeine for acute postoperative pain in adults. Two review authors independently considered trials for inclusion in the review, assessed risk of bias, and extracted data. We used the area under the pain relief versus time curve to derive the proportion of participants with at least 50% pain relief over six hours prescribed either ibuprofen plus caffeine or placebo. We calculated the risk ratio (RR) and number needed to treat to benefit (NNT). We used information on the use of rescue medication to calculate the proportion of participants requiring rescue medication and the weighted mean of the median time to use. We also collected information on adverse effects. We identified five randomised, double-blind studies with 1501 participants, but only four had been published and had relevant outcome data. These four studies were of high quality, although two of the studies were small.Both ibuprofen 200 mg + caffeine 100 mg and ibuprofen 100 mg + caffeine 100 mg produced significantly more participants than placebo who achieved at least 50% of maximum pain relief over six hours, and both doses significantly reduced remedication rates (moderate quality evidence). For at least 50% of maximum pain relief, the NNT was 2.1 (95% confidence interval 1.8 to 2.5) for ibuprofen 200 mg + caffeine 100 mg (four studies, 334 participants) and 2.4 (1.9 to 3.1) for ibuprofen 100 mg + caffeine 100 mg (two studies, 200 participants) (moderate quality evidence). These values were close to those predicted by published models for combination analgesics in acute pain, and were supported by low (good) NNT values for prevention of remedication.Adverse event rates were low, and no sensible analysis was possible. For ibuprofen 200 mg + caffeine 100 mg particularly, the low NNT value is among the lowest (best) values for analgesics in this pain model. The combination is not commonly available, but can be probably be achieved by taking a single 200 mg ibuprofen tablet with a cup of modestly strong coffee or caffeine tablets. In principle, this can deliver good analgesia at lower doses of ibuprofen.


Telehealth in Plastic Surgery: A Veterans Affairs Hospital Perspective.
Telemedicine is a rapidly growing tool since its invention in the 1950s. Recently, it has expanded to the field of plastic surgery. In the Connecticut VA System, there is 1 plastic surgeon at a central location for the state of Connecticut and southern Massachusetts. Our aim was to pilot a telehealth program for plastic surgery consultation within the VA to improve access to subspecialty care. We intend to discuss the value of telehealth as part of consultation services, and assess patient attitudes toward telemedicine. Patients in the Connecticut VA System referred for plastic surgery consultation for evaluation of nonurgent diagnoses, such as skin lesions, carpal tunnel syndrome, and chronic wounds, were invited to participate. After being appropriately consented according to Connecticut state law, patients completed a postvisit questionnaire rating their overall satisfaction, quality of interaction, and ability to communicate using a modified 10-point Likert scale. Means were calculated for numerical responses. Preference for future telehealth visits was reported as a percentage. Forty-one of 44 eligible patients elected to participate. Of those patients, 83% (n = 34) stated they prefer telemedicine services for future visits. Patients rated overall satisfaction 9.2/10, overall quality of interaction 9.2/10, ability to communicate 9.3/10, and sound and video quality 8.6/10 and 9.0/10, respectively. Remote video telemedicine is a feasible method of plastic surgery consultation and results in high patient satisfaction. Further studies should focus on cost-effectiveness and ways to broaden the use of telehealth services in plastic surgery.


[The antiblastic properties of reumycin (experimental research)].
The work deals with the results of study of the antineoplastic activity of a Soviet produced antineoplastic antibiotic reumycin in relation to 35 glial tumors of various degrees of malignancy in primary cultures and in heterotransplantation under the capsule of kidneys in mice. In tissue cultures 71.5% of gliomas were sensitive to reumycin, in the subcapsular test 75% of tumors proved to be reumycin sensitive with fluctuations of inhibited growth of heterotransplants within a range of 27-75%. Among the reumycin resistant tumors, the cultures of one of the glioblastomas showed stimulation of proliferation, which justifies the necessity for testing the individual sensitivity of the tumor to reumycin in each concrete patient.


[Biochemical parameters of blood and morpho-functional state of the liver of experimental animals by the actions of lead sulfide nanoparticles in different time study].
Biochemical studies revealed that alanine aminotransferase levels changing first during short action (30 injections) of lead sulfide nanoparticles of size 10 and 30 nm, and the ionic form of a 400 nm lead while the growth of both enzymes (aspartate aminotransferase and alanine aminotransferase) activity during long-term exposure (60 injections) is the same intensity. It it confirmed by the value of de Ritis coefficient, which is statistically the same as control. Morphological studies also confirm these data--degenerative changes of hepatocytes, reactive changes of the stroma and vascular responses were detected. It is shown that the severity of metabolic and morphological damages in the liver increased with prolonging the duration of lead nanoparticles intake.


Disparities in imaging utilization for acute ischemic stroke based on patient insurance status.
Previous studies have shown socioeconomic disparities in imaging utilization for both acute and chronic diseases. We studied a nationwide database to determine whether insurance-based disparities exist in the utilization of imaging for acute ischemic stroke. Inpatients with a primary diagnosis of acute ischemic stroke from November 2005 through December 2011 were identified from the Perspective database. Patients were stratified into four groups according to insurance status as follows: uninsured, Medicaid, Medicare, and private insurance. Utilization rates of head CT, perfusion CT, head MRI, noninvasive head angiography (including head CT angiography [CTA] and head MR angiography [MRA]), noninvasive neck angiography (including neck CTA and neck MRA), carotid ultrasound, and echocardiography were compared using a chi-square test. A multivariable logistic regression model adjusting for potential confounding variables was fit to determine the association between insurance status and imaging utilization. A total of 210,212 patients were included in this study: 10,396 patients (5.0%) were uninsured, 14,243 patients (6.8%) had Medicaid, 153,209 patients (72.9%) had Medicare, and 32,364 patients (15.4%) had private insurance. Even after we had controlled for confounding variables, significant disparities existed in imaging utilization. Compared with patients with private insurance, uninsured patients had significantly lower odds of noninvasive head angiography (odds ratio [OR] = 0.78, 95% CI = 0.74-0.81, p < 0.0001), neck angiography (OR = 0.79, 95% CI = 0.76-0.83, p < 0.0001), and head MRI (OR = 0.77, 95% CI = 0.74-0.81, p < 0.0001). The same was true for Medicaid and Medicare patients. Disparities exist in the utilization of noninvasive head and neck imaging, MRI, and echocardiography for patients with acute ischemic stroke based on patient insurance status. More research is needed to understand these disparities.


Purinergic receptor immunoreactivity in the rostral ventromedial medulla.
The rostral ventromedial medulla (RVM) has long been recognized to play a pivotal role in nociceptive modulation. Pro-nociception within the RVM is associated with a distinct functional class of neurons, ON-cells that begin to discharge immediately before nocifensive reflexes. Anti-nociceptive function within the RVM, including the analgesic response to opiates, is associated with another distinct class, OFF-cells, which pause immediately prior to nocifensive reflexes. A third class of RVM neurons, NEUTRAL-cells, does not alter firing in association with nocifensive reflexes. ON-, OFF- and NEUTRAL-cells show differential responsiveness to various behaviorally relevant neuromodulators, including purinergic ligands. Iontophoresis of semi-selective P2X ligands, which are associated with nociceptive transmission in the spinal cord and dorsal root ganglia, preferentially activate ON-cells. By contrast, P2Y ligands activate OFF-cells and P1 ligands suppress the firing of NEUTRAL cells. The current study investigates the distribution of P2X, P2Y and P1 receptor immunoreactivity in RVM neurons of Sprague-Dawley rats. Co-localization with tryptophan hydroxylase (TPH), a well-established marker for serotonergic neurons was also studied. Immunoreactivity for the four purinergic receptor subtypes examined was abundant in all anatomical subdivisions of the RVM. By contrast, TPH-immunoreactivity was restricted to a relatively small subset of RVM neurons concentrated in the nucleus raphe magnus and pallidus, as expected. There was a significant degree of co-localization of each purinergic receptor subtype with TPH-immunoreactivity. This co-localization was most pronounced for P2Y1 receptor immunoreactivity, although this was the least abundant among the different purinergic receptor subtypes examined. Immunoreactivity for multiple purinergic receptor subtypes was often co-localized in single neurons. These results confirm the physiological finding that purinergic receptors are widely expressed in the RVM. Purinergic neurotransmission in this region may play an important role in nociception and/or nociceptive modulation, as at other levels of the neuraxis.


Acetylcholine and bradykinin relax intrapulmonary arteries by acting on endothelial cells: role in lung vascular diseases.
Acetylcholine and bradykinin produced potent relaxation of isolated canine intrapulmonary arteries contracted by serotonin, norepinephrine, or phenylephrine-provided the endothelium was left intact. Selective mechanical destruction of the endothelium transformed the activity of these substances from vasodilatation to vasoconstriction. Acetylcholine-induced relaxations, in the presence of intact endothelium, could be selectively inhibited competitively by atropine, but could not be inhibited by cyclooxygenase inhibitors, a lipoxygenase inhibitor, adrenergic blocking drugs, or histaminergic antagonists. RElaxations produced by prostacyclin, prostaglandin E1, isoproterenol, papaverine, or histamine H2-receptor agonists were not modified, or attenuated, by selective destruction of pulmonary endothelial cells. These observations might provide insight into the etiology of the increased pulmonary resistance observed in pulmonary hypertension and shock lung.


Drugs to cure avian influenza infection--multiple ways to prevent cell death.
New treatments and new drugs for avian influenza virus (AIV) infection are developed continually, but there are still high mortality rates. The main reason may be that not all cell death pathways induced by AIV were blocked by the current therapies. In this review, drugs for AIV and associated acute respiratory distress syndrome (ARDS) are summarized. The roles of antioxidant (vitamin C) and multiple immunomodulators (such as Celecoxib, Mesalazine and Eritoran) are discussed. The clinical care of ARDS may result in ischemia reperfusion injury to poorly ventilated alveolar cells. Cyclosporin A should effectively inhibit this kind of damages and, therefore, may be the key drug for the survival of patients with virus-induced ARDS. Treatment with protease inhibitor Ulinastatin could also protect lysosome integrity after the infection. Through these analyses, a large drug combination is proposed, which may hypothetically greatly reduce the mortality rate.


Fully automatic detection of lung nodules in CT images using a hybrid feature set.
The aim of this study was to develop a novel technique for lung nodule detection using an optimized feature set. This feature set has been achieved after rigorous experimentation, which has helped in reducing the false positives significantly. The proposed method starts with preprocessing, removing any present noise from input images, followed by lung segmentation using optimal thresholding. Then the image is enhanced using multiscale dot enhancement filtering prior to nodule detection and feature extraction. Finally, classification of lung nodules is achieved using Support Vector Machine (SVM) classifier. The feature set consists of intensity, shape (2D and 3D) and texture features, which have been selected to optimize the sensitivity and reduce false positives. In addition to SVM, some other supervised classifiers like K-Nearest-Neighbor (KNN), Decision Tree and Linear Discriminant Analysis (LDA) have also been used for performance comparison. The extracted features have also been compared class-wise to determine the most relevant features for lung nodule detection. The proposed system has been evaluated using 850 scans from Lung Image Database Consortium (LIDC) dataset and k-fold cross-validation scheme. The overall sensitivity has been improved compared to the previous methods and false positives per scan have been reduced significantly. The achieved sensitivities at detection and classification stages are 94.20% and 98.15%, respectively, with only 2.19 false positives per scan. It is very difficult to achieve high performance metrics using only a single feature class therefore hybrid approach in feature selection remains a better choice. Choosing right set of features can improve the overall accuracy of the system by improving the sensitivity and reducing false positives.


CT diagnosis of internal mammary artery injury caused by blunt trauma.
To describe the computed tomography (CT) findings associated with active bleeding from the internal mammary artery (IMA) in blunt trauma victims and to assess complications related to IMA haemorrhage. All cases of active IMA haemorrhage identified in blunt trauma patients on admission CT were identified from a trauma radiology data base covering 1990-1999. Computed tomography examinations, operative and medical records were reviewed to ascertain CT findings, complications, and patient outcome. The determination of active bleeding required CT evidence of a central contrast blush of CT density within 10 HU of an adjacent artery surrounded by haematoma. Four patients with CT evidence of active IMA haemorrhage were identified. All cases had surgical confirmation of an IMA source of haemorrhage. There were three patients with unilateral and one patient with bilateral IMA disruption. Three patients exhibited clinical signs of cardiac tamponade related to compression of one or more cardiac chambers by the anterior mediastinal haematoma. Sudden clinical deterioration compatible with tamponade developed in all three patients. Early CT recognition of active bleeding within the chest can direct rapid surgical or angiographic intervention. On-going blood loss and, in particular, the threat of cardiac tamponade must be considered with IMA injury.


Voltammetric determination of polyphenolic content in pomegranate juice using a poly(gallic acid)/multiwalled carbon nanotube modified electrode.
A simple and sensitive poly(gallic acid)/multiwalled carbon nanotube modified glassy carbon electrode (PGA/MWCNT/GCE) electrochemical sensor was prepared for direct determination of the total phenolic content (TPC) as gallic acid equivalent. The GCE working electrode was electrochemically modified and characterized using scanning electron microscope (SEM), cyclic voltammetry (CV), chronoamperometry and chronocoulometry. It was found that gallic acid (GA) exhibits a superior electrochemical response on the PGA/MWCNT/GCE sensor in comparison with bare GCE. The results reveal that a PGA/MWCNT/GCE sensor can remarkably enhance the electro-oxidation signal of GA as well as shift the peak potentials towards less positive potential values. The dependence of peak current on accumulation potential, accumulation time and pH were investigated by square-wave voltammetry (SWV) to optimize the experimental conditions for the determination of GA. Using the optimized conditions, the sensor responded linearly to a GA concentration throughout the range of 4.97 × 10(-6) to 3.38 × 10(-5) M with a detection limit of 3.22 × 10(-6) M (S/N = 3). The fabricated sensor shows good selectivity, stability, repeatability and (101%) recovery. The sensor was successfully utilized for the determination of total phenolic content in fresh pomegranate juice without interference of ascorbic acid, fructose, potassium nitrate and barbituric acid. The obtained data were compared with the standard Folin-Ciocalteu spectrophotometric results.


[Exudative onycholysis and acute bacterial paronychia related to BIBF-1120 and paclitaxel: response to topical therapy].
A case of a 50 years-old breast cancer patient treated with weekly paclitaxel and BIBF 1120 is reported herein. At the end of the twelfth cycle of chemotherapy, the patient developed distal onycholysis with intense hyponychium serous exudates, pain and malodor in all her fingernails. It was treated with topical fusidic acid and 1% methylprednisolone aceponate two times daily, with an excellent clinical response from the first three days of treatment. Bacterial paronychia with nail plate loss of the fifth left fingernail was observed a week after the topical therapy was started, with positive cultures for Methicillin susceptible Staphylococcus aureus. There are few reported cases of exudative onycholysis associated with chemotherapy. However, these are especially related to paclitaxel. No recurrences of nail disturbances were observed weeks after the end of chemotherapy. Topical corticosteroids and fusidic acid could be considered as a therapeutic option when exudative onycholysis related to paclitaxel is established


Near-atomic resolution using electron cryomicroscopy and single-particle reconstruction.
Electron cryomicroscopy (cryo-EM) yields images of macromolecular assemblies and their components, from which 3D structures can be determined, by using an image processing method commonly known as "single-particle reconstruction." During the past two decades, this technique has become an important tool for 3D structure determination, but it generally has not been possible to determine atomic models. In principle, individual molecular images contain high-resolution information contaminated by a much higher level of noise. In practice, it has been unclear whether current averaging methods are adequate to extract this information from the background. We present here a reconstruction, obtained by using recently developed image processing methods, of the rotavirus inner capsid particle ("double-layer particle" or DLP) at a resolution suitable for interpretation by an atomic model. The result establishes single-particle reconstruction as a high-resolution technique. We show by direct comparison that the cryo-EM reconstruction of viral protein 6 (VP6) of the rotavirus DLP is similar in clarity to a 3.8-A resolution map obtained from x-ray crystallography. At this resolution, most of the amino acid side chains produce recognizable density. The icosahedral symmetry of the particle was an important factor in achieving this resolution in the cryo-EM analysis, but as the size of recordable datasets increases, single-particle reconstruction also is likely to yield structures at comparable resolution from samples of much lower symmetry. This potential has broad implications for structural cell biology.


Morphological and Chemical Profile of Three Tomato (Solanum lycopersicum L.) Landraces of A Semi-Arid Mediterranean Environment.
Puglia (Southern Italy), particularly rich in tomato agro-biodiversity, can be considered a typical region of the semi-arid Mediterranean environments. In this study, three local varieties of tomato (Manduria, Giallo di Crispiano and Regina) were characterized by using morphological descriptors according to international standards. Chemical (isoprenoids, ascorbic acid, total phenols, sugars and mineral content) and agronomic assessment were carried out to highlight the specific traits of these local varieties well adapted to a semi-arid environment. Data of morphological traits according to the "International Union for the Protection of New Varieties of Plants" (UPOV) guideline evidenced a clear distinctness among all three landraces, especially as regards fruits. Results also highlighted that a great part of variation in chemical traits was almost exclusively due to genotypes, while in a few cases observed differences resulted from the interaction between genotype and harvest time. The results of the present study may represent the first step toward the recognition of "conservation variety" status for Regina, Giallo di Crispiano and Manduria tomato landraces. At the same time, both quality traits and agronomic performance of these tomato genotypes suggest the possibility of their cultivation in other semi-arid environments also considering their quality traits, in view of a sustainable production.


The Effect of Fullerenol C60(OH)36 on the Antioxidant Defense System in Erythrocytes.
Background: Fullerenols (water-soluble derivatives of fullerenes), such as C60(OH)36, are biocompatible molecules with a high ability to scavenge reactive oxygen species (ROS), but the mechanism of their antioxidant action and cooperation with endogenous redox machinery remains unrecognized. Fullerenols rapidly distribute through blood cells; therefore, we investigated the effect of C60(OH)36 on the antioxidant defense system in erythrocytes during their prolonged incubation. Methods: Human erythrocytes were treated with fullerenol at concentrations of 50-150 µg/mL, incubated for 3 and 48 h at 37 °C, and then hemolyzed. The level of oxidative stress was determined by examining the level of thiol groups, the activity of antioxidant enzymes (catalase, glutathione peroxidase, glutathione reductase, and glutathione transferase), and by measuring erythrocyte microviscosity. Results: The level of thiol groups in stored erythrocytes decreased; however, in the presence of higher concentrations of C60(OH)36 (100 and 150 µg/mL), the level of -SH groups increased compared to the control. Extending the incubation to 48 h caused a decrease in antioxidant enzyme activity, but the addition of fullerenol, especially at higher concentrations (100-150 µg/mL), increased its activity. We observed that C60(OH)36 had no effect on the microviscosity of the interior of the erythrocytes. Conclusions: In conclusion, our results indicated that water-soluble C60(OH)36 has antioxidant potential and efficiently supports the enzymatic antioxidant system within the cell. These effects are probably related to the direct interaction of C60(OH)36 with the enzyme that causes its structural changes.


[The first appearance of Meckel's cartilage in the fetus].
Meckel's cartilage plays an important role in the topographical organisation and in the differentiation of the facial structure during the embryonal and even much later during the foetal period. Our observations on serial sections carried out in two human foetuses aged 12 and 16 weeks indicate that the two dorsal (tympanic) and ventral (mandibular) branches of Meckel's cartilage are perfectly defined at 16 weeks. In the dorsal branch, the primordia of the incus and of head of the malleus are still composed on non-ossified cartilage. In the ventral branch, it is also possible to describe at 16 weeks three posterior, medial and anterior parts which are composed of cartilage. The initiating role played by the ventral part of Meckel's cartilage on the ossification of the mandible leads during the embryonal period to the formation of the mandibular primary growth center, which is therefore clearly defined in our first stage at 12 weeks. The partial fibrous evolution and the regression of the major part of the ventral branch of Meckel's cartilage only start after 16 weeks of intrauterine life.


Contribution of parvalbumin and somatostatin-expressing GABAergic neurons to slow oscillations and the balance in beta-gamma oscillations across cortical layers.
Cortical interneurons are classified into several subtypes that contribute to cortical oscillatory activity. Parvalbumin (PV)-expressing cells, a type of inhibitory interneuron, are involved in the gamma oscillations of local field potentials (LFPs). Under ketamine-xylazine anesthesia or sleep, mammalian cortical circuits exhibit slow oscillations in which the active-up state and silent-down state alternate at ~1 Hz. The up state is composed of various high-frequency oscillations, including gamma oscillations. However, it is unclear how PV cells and somatostatin (SOM) cells contribute to the slow oscillations and the high-frequency oscillations nested in the up state. To address these questions, we used mice lacking glutamate decarboxylase 67, primarily in PV cells (PV-GAD67 mice) or in SOM cells (SOM-GAD67 mice). We then compared LFPs between PV-GAD67 mice and SOM-GAD67 mice. PV cells target the proximal regions of pyramidal cells, whereas SOM cells are dendrite-preferring interneurons. We found that the up state was shortened in duration in the PV-GAD67 mice, but tended to be longer in SOM-GAD67 mice. Firing rate tended to increase in PV-GAD67 mice, but tended to decrease in SOM-GAD67 mice. We also found that delta oscillations tended to increase in SOM-GAD67 mice, but tended to decrease in PV-GAD67 mice. Current source density and wavelet analyses were performed to determine the depth profiles of various high-frequency oscillations. High gamma and ripple (60-200 Hz) power decreased in the neocortical upper layers specifically in PV-GAD67 mice, but not in SOM-GAD67. In addition, beta power (15-30 Hz) increased in the deep layers, specifically in PV-GAD67 mice. These results suggest that PV cells play important roles in persistence of the up state and in the balance between gamma and beta bands across cortical layers, whereas SOM and PV cells may make an asymmetric contribution to regulate up-state and delta oscillations.


Daily vs twice daily enoxaparin in the prevention of venous thromboembolic disorders during rehabilitation following acute spinal cord injury.
Subcutaneous administration of low molecular weight heparin (eg, enoxaparin) has been shown to be safe and effective in the prevention of acute venous thromboembolic (TE) disease following acute spinal cord injury (SCI) in the rehabilitation setting. However, emerging evidence suggests that different dosing strategies may be equivalent. To determine whether subcutaneous enoxaparin, 40 mg once daily, when compared with subcutaneous enoxaparin, 30 mg twice daily, is equally safe and effective in the prevention of venous TE disease in patients with SCI. Retrospective chart review. A freestanding, inpatient, acute, rehabilitation hospital. One hundred and twenty-nine patients admitted from June 2000 through June 2002 for inpatient rehabilitation following an acute SCI who received either enoxaparin, 40 mg once daily, or enoxaparin, 30 mg twice daily, for prophylaxis for TE disease. Equivalent prophylaxis efficacy was seen in both enoxaparin groups. Symptomatic venous thromboembolism did not differ, with deep vein thromboses occurring in 1 of 49 (2.0%) patients receiving twice-daily enoxaparin, and 1 of 80 (1.25%) patients receiving once-daily enoxaparin (chi2 = 0.125, NS). Pulmonary embolism was seen in 1 of 49 (2.0%) patients treated with twice-daily enoxaparin and in none of the patients in the once-daily group (chi2 = 1.64, NS). Bleeding complications also did not differ between the 2 treatment groups; these were observed in 2 of 49 (4.1%) patients receiving twice-daily enoxaparin and in 5 of 80 (6.3%) patients receiving once-daily enoxaparin (chi2 = 0.228, NS). Subcutaneous enoxaparin administered once or twice daily is equally effective for the prevention of venous TE disease. Both dosing strategies are associated with a low incidence of bleeding in patients with SCI who are undergoing rehabilitation.


Expression of p27 and p53: comparative analysis of uterine carcinosarcoma and endometrial carcinoma.
The aim of the study was to assess both p27 and p53 expression in the stromal and epithelial component of carcinosarcoma and to assess if their expression in the latter is different than in endometrial carcinoma. Immunohistochemical staining for p27 and p53 was performed on paraffin-embedded tissue blocks of 18 uterine specimens with carcinosarcoma and their expression assessed. Their expression in the epithelial element was also compared to that in 35 paraffin-embedded tissue blocks of endometrial endometrioid carcinoma. Reduced p27 expression was observed in a similarly high proportion of the stromal (77.8%) as well as of the epithelial component (66.7%) of carcinosarcoma. Although statistically not significant, the proportion of reduced p27 expression in endometrial carcinoma (85.7%) was higher than in the epithelial element of carcinosarcoma. The percentage of p53 overexpression in both elements of carcinosarcomas and in endometrial carcinomas was low and also similar (27.8 and 20.0%, respectively). Our results indicate that reduced p27 expression is common and p53 overexpression is infrequent in carcinosarcoma. Their similar rates of expression in the stromal and epithelial elements of the tumor support the contention of a monoclonal origin of carcinosarcoma. Unexpectedly, reduced p27 expression is more common in endometrial carcinoma than in the epithelial element of carcinosarcoma, in spite of the less favorable prognosticators and outcome in the latter. Further studies of p27 expression in carcinosarcoma are indicated to establish its clinical value in this aggressive malignancy.


Middle-Range Theory: Coping and Adaptation with Active Aging.
Various disciplines focus on a multiplicity of aspects of aging: lifestyles, personal biological factors, psychological conditions, health conditions, physical environment, and social and economic factors. The aforementioned are all related to the determinants of active aging. The aim is to describe the development of a middle-range theory based on coping and adaptation with active aging. Concepts and relationships derived from Roy's model of adaptation are included. The proposed concepts are hope, health habits, coping with aging, social relations, and active aging.


THz and mid-IR spectroscopy of interstellar ice analogs: methyl and carboxylic acid groups.
A fundamental problem in astrochemistry concerns the synthesis and survival of complex organic molecules (COMs) throughout the process of star and planet formation. While it is generally accepted that most complex molecules and prebiotic species form in the solid phase on icy grain particles, a complete understanding of the formation pathways is still largely lacking. To take full advantage of the enormous number of available THz observations (e.g., Herschel Space Observatory, SOFIA, and ALMA), laboratory analogs must be studied systematically. Here, we present the THz (0.3-7.5 THz; 10-250 cm(-1)) and mid-IR (400-4000 cm(-1)) spectra of astrophysically-relevant species that share the same functional groups, including formic acid (HCOOH) and acetic acid (CH3COOH), and acetaldehyde (CH3CHO) and acetone ((CH3)2CO), compared to more abundant interstellar molecules such as water (H2O), methanol (CH3OH), and carbon monoxide (CO). A suite of pure and mixed binary ices are discussed. The effects on the spectra due to the composition and the structure of the ice at different temperatures are shown. Our results demonstrate that THz spectra are sensitive to reversible and irreversible transformations within the ice caused by thermal processing, suggesting that THz spectra can be used to study the composition, structure, and thermal history of interstellar ices. Moreover, the THz spectrum of an individual species depends on the functional group(s) within that molecule. Thus, future THz studies of different functional groups will help in characterizing the chemistry and physics of the interstellar medium (ISM).


Student reactions to a community-based experience for health professions education.
Student responses to a community health program in a rural Mexican-American town are presented. Off-campus training was provided for 104 health professions students, during which health evaluations were made of 741 elementary school children. The students reported that the program allowed them to apply clinical skills, that they were more interested in rural practice, and that their reaction to working in teams was overwhelmingly positive. The students reported that teaming offered as advantages greater knowledge and appreciation of other disciplines, more comprehensive patient care, increased knowledge of communication processes, and exposure to relevant clinical skills from other disciplines. Their perceptions of the advantages of teaming varied among the disciplines. The program provided a learning experience for the students and resulted in improved health care for the town.


Influence of various superhydrophilic treatments of titanium on the initial attachment, proliferation, and differentiation of osteoblast-like cells.
The purpose of this study was to investigate the influence of superhydrophilic treatments of titanium on the behavior of osteoblastlike cells. Superhydrophilic specimens were prepared with sandblast and acid-etching (DW), oxygen plasma (Plasma) and ultraviolet light (UV), and were stored in distilled water for 3 days immediately after these treatments. Specimens stored in air for 3 weeks were used as a control Air group. Initial cell attachment, proliferation, alkaline phosphatase activity, and osteocalcin secretion of mouse osteoblast-like cells MC3T3-E1 were enhanced more on superhydrophilic groups than were Air specimens. On confocal laser scanning microscope images of cell morphology, the expression of actin filaments was observed on the superhydrophilic groups, whereas relatively little actin filament expression was seen on the Air surfaces on all culture periods. These results indicate that DW, Plasma, or UV treatment has potential for the creation and maintenance of superhydrophilic surfaces and the enhancement of the initial attachment, proliferation, and differentiation of osteoblast-like cells.


Stereo disparity facilitates view generalization during shape recognition for solid multipart objects.
Current theories of object recognition in human vision make different predictions about whether the recognition of complex, multipart objects should be influenced by shape information about surface depth orientation and curvature derived from stereo disparity. We examined this issue in five experiments using a recognition memory paradigm in which observers (N = 134) memorized and then discriminated sets of 3D novel objects at trained and untrained viewpoints under either mono or stereo viewing conditions. In order to explore the conditions under which stereo-defined shape information contributes to object recognition we systematically varied the difficulty of view generalization by increasing the angular disparity between trained and untrained views. In one series of experiments, objects were presented from either previously trained views or untrained views rotated (15°, 30°, or 60°) along the same plane. In separate experiments we examined whether view generalization effects interacted with the vertical or horizontal plane of object rotation across 40° viewpoint changes. The results showed robust viewpoint-dependent performance costs: Observers were more efficient in recognizing learned objects from trained than from untrained views, and recognition was worse for extrapolated than for interpolated untrained views. We also found that performance was enhanced by stereo viewing but only at larger angular disparities between trained and untrained views. These findings show that object recognition is not based solely on 2D image information but that it can be facilitated by shape information derived from stereo disparity.


WOX2 and STIMPY-LIKE/WOX8 promote cotyledon boundary formation in Arabidopsis.
One of the key events in dicot plant embryogenesis is the emergence of the two cotyledon primordia, which marks the transition from radial symmetry to bilateral symmetry. In Arabidopsis thaliana, the three CUP-SHAPED COTYLEDON (CUC) genes are responsible for determining the boundary region between the cotyledons. However, the mechanisms controlling their transcription activation are not well understood. Previous studies found that several WOX family homeobox transcription factors are involved in embryo apical patterning and cotyledon development. Here we show that WOX2 and STIMPY-LIKE (STPL/WOX8) act redundantly to differentially regulate the expression of the CUC genes in promoting the establishment of the cotyledon boundary, without affecting the primary shoot meristem. Loss of both WOX2 and STPL results in reduced CUC2 and CUC3 expression in one side of the embryo, but an expansion of the CUC1 domain. Furthermore, we found that STPL is expressed in the embryo proper, and its activation is enhanced by the removal of WOX2, providing an explanation for the functional redundancy between WOX2 and STPL. Additional evidence also showed that WOX2 and STPL function independently in regulating different aspects of local auxin gradient formation during early embryogenesis.


Implant-based breast reconstruction with allograft.
Reconstructive breast surgeons often prefer autologous tissue for those patients who can tolerate it. However, reconstruction with implants is technically easier, quicker, less traumatic, and less costly. Nevertheless, inherent problems associated with implants present some limitations to their use in reconstruction. In this study, a multipurpose application of allogenic dermal grafts to solve the problems seen in immediate breast reconstruction with implants was examined. After skin-sparing mastectomy, a subpectoral pocket is created for the implant. After the origins of the pectoralis major muscle are released, AlloDerm is sewn to the lower pole of the released muscle. Grafts are tailored according to the dimensions of the defect. AlloDerm, an allogenic dermal graft, is fixed to the pectoralis major superiorly, to the chest wall inferiorly and inferomedially, and to the serratus anterior flap laterally. A postoperative adjustable implant is placed into the pocket that has bene created. Allografts were used to reconstruct 30 breasts in 24 patients immediately after skin- or nipple-sparing mastectomy. There was no complication related to simultaneous allograft application. No rippling, symmastia, or bottoming-out deformity was seen. Breast reconstruction with implant and AlloDerm enables the surgeon to create a submuscular pocket large enough to place a fully inflated breast implant. Interposition of a lower-pole internal sling, to increase the capacity of the pectoralis major pocket while providing immediate ability to achieve full or subtotal prosthetic fill and optimal aesthetic breast contouring and to maintain thicker muscle coverage in the upper and medial pole areas and stronger lower pole support, would obviate many of the current disadvantages to implant reconstruction.


Newer molecular approaches to detect known viruses and previously unrecognized viruses in cell substrates.
The theme of this brief presentation is that there are tremendous advances not only in PCR-based detection but in other novel ways of detecting viruses and other agents. Much of the research is now being driven by the bio-defence agenda. Somehow those with manufacturing and regulatory responsibilities have to keep abreast of these developments and incorporate them as they become available into defining the safety of cell substrates. I am concerned that these techniques are and may remain academic research tools without validation or standardization, and that individual academic laboratories will not develop the capacity and special skills necessary to satisfy regulatory requirements, but that there is a tremendous research incentive in this area that needs to be linked with the development of safe prophylactic and therapeutic biologicals. With that, I will finish what I would consider as a report rather than an original talk.


Genomics and microarray for detection and diagnostics.
Genomics provided biomedical scientists an inventory of all genes and sequences present in a living being. This provides an unique opportunity to the scientists to predict and study biological functions of these genes. The changes in the gene expression regulated by genomic sequences therefore reflect changes in the molecular processes working in a cell or tissue in response to external factors including exposure to toxic compounds and pathogens. Microarray offers a biotechnological revolution with the help of DNA chemistry, silicon chip technology and optics to be used to monitor gene expression for thousands of genes in one single experiment. Briefly, 20,000 to 100,000 unique DNA molecules get applied by a robot to the surface of silicon wafers (approximately the size of a microscope slide). Using a single microarray experiment, the expression level of 20,000 to 100,000 genes will be examined in one single experiment. Genomics and microarray have a significant role and impact on the design and development of modern detection and diagnostic tools in several different ways. Microarray tools are now used on regular basis for monitoring gene expression of large number of genes and also frequently applied to DNA sequence analysis, immunology, genotyping, and molecular diagnosing. For diagnostics, these tools can be used to distinguish and differentiate between different DNA fragments that differ by as little as a single nucleotide polymorphism (SNP). These microarrays can be divided based on the gene density spots that will be high density (>10,000 spots) per slide, medium (< 1000 > 100) and low density (< 100). High-density arrays have proven to be very useful in disease diagnosis especially in diagnosis and classification of different types of cancers. These microarray tools hold tremendous potential for pathogen detection, which will be comprised, of unique sets of genes (also referred to as "signatures") able to unambiguously identify the species and strain of pathogens of interest.


Geographical relationships between sociodemographic factors and incidence of cervical cancer in the Netherlands 1989-2003.
In many industrialized countries, with some degree of screening, cervical cancer nowadays is most frequent among women of lower socioeconomic status (SES), partly owing to their lower participation in screening. This study aims to provide support for specification of mass screening policy for cervical cancer by describing relationships between sociodemographic factors and the incidence of cervical cancer in the Netherlands based on geographical differences and by analysing the relationship between SES of neighbourhood and individual tumour characteristics. Municipality-specific, age-adjusted incidence rates for cervical cancer were calculated from the Netherlands Cancer Registry, and data on sociodemographic factors were obtained from Statistics Netherlands. Logistic regression analysis was performed to investigate determinants of variations in incidence at the ecological level. An additional analysis linked individual tumour characteristics to SES estimates at the postal code level by calculating relative risks (RR). The incidence was higher in municipalities with a high prevalence of immigrants [odds ratios 7.9, 1.4-47 95% confidence intervals (CI)] and with more individuals on welfare (odds ratios 8.6, 1.7-43 95% CI). Patients residing in neighbourhoods with lower SES had higher Federation of Gynecology and Obstetrics stages (RR 1.4, 1.2-1.6 95% CI) and fewer adenocarcinomas (RR 0.7, 0.6-0.9 95% CI), and were younger at diagnosis (P<0.001). Cervical cancer is more common among women of lower SES and immigrant women. This, together with the finding that lower SES is associated with more advanced cancer and consequently worse survival, emphasizes the importance of future cervical cancer prevention programmes targeted at women of lower SES who do not participate in opportunistic screening.


Anti-Obesity Effects of Artemisia annua Extract in Zucker Fatty Rats and High-Fat Diet Sprague Dawley Rats through Upregulation of Uncoupling Protein 1.
Obesity is a widespread disease and is caused mainly by excessive adipocyte differentiation and fat accumulation. Peroxisome proliferation-activated receptor γ (PPARγ) and CCAAT/enhancer-binding proteins (C/EBP) are major components for regulating adipocyte differentiation. Uncoupling protein 1 (UCP1) is a transmembrane protein that can convert white fat to brown adipose tissue. Artemisia annua L. has long been used in East Asia as an herbal drug for anti-oxidant, anti-bacterial, and anti-obesity purposes. We investigated the effects of water extracts of A. annua (WEAA) in C3H10T1/2, a mesenchymal stem cell line, by measuring the level of intracellular fat accumulation and the expression of genes associated with adipocyte differentiation. We also evaluated anti-obesity effects of WEAA in Zucker rats, a genetic model for the study of obesity, and in Sprague Dawley rats with high-fat diet (HFD)-induced obesity. In this study, WEAA reduced the expression levels of PPARγ and C/EBPα in C3H10T1/2 cells, as well as the expression of enzymes that regulate fatty acid metabolism. In the Zucker fatty rat model and the HFD-induced obesity rat model, WEAA significantly decreased adipogenic differentiation and white fat accumulation between the scapulae, in contrast to the brown fat that remained unchanged between the groups. A. annua suppressed the expression of the adipocyte differentiation-promoting genes, while increasing the expression of UCP1. These results indicated that WEAA could reduce adipocyte differentiation and fat accumulation in in vitro and in vivo model systems, resulting in suppression of obesity and the occurrence of fatty liver due to a HFD.


Nosocomial transmission of C. difficile in English hospitals from patients with symptomatic infection.
Recent evidence suggests that less than one-quarter of patients with symptomatic nosocomial Clostridium difficile infections (CDI) are linked to other in-patients. However, this evidence was limited to one geographic area. We aimed to investigate the level of symptomatic CDI transmission in hospitals located across England from 2008 to 2012. A generalized additive mixed-effects Poisson model was fitted to English hospital-surveillance data. After adjusting for seasonal fluctuations and between-hospital variation in reported CDI over time, possible clustering (transmission between symptomatic in-patients) of CDI cases was identified. We hypothesised that a temporal proximity would be reflected in the degree of correlation between in-hospital CDI cases per week. This correlation was modelled through a latent autoregressive structure of order 1 (AR(1)). Forty-six hospitals (33 general, seven specialist, and six teaching hospitals) located in all English regions met our criteria. In total, 12,717 CDI cases were identified; seventy-five per cent of these occurred >48 hours after admission. There were slight increases in reports during winter months. We found a low, but statistically significant, correlation between successive weekly CDI case incidences (phi = 0.029, 95%CI: 0.009-0.049). This correlation was five times stronger in a subgroup analysis restricted to teaching hospitals (phi = 0.104, 95%CI: 0.048-0.159). The results suggest that symptomatic patient-to-patient transmission has been a source of CDI-acquisition in English hospitals in recent years, and that this might be a more important transmission route in teaching hospitals. Nonetheless, the weak correlation indicates that, in line with recent evidence, symptomatic cases might not be the primary source of nosocomial CDI in England.


Effect of sweet chestnut tannin (SCT) on the performance, microbial status of intestine and histological characteristics of intestine wall in chickens.
1. In experiments carried out with 950 one-day-old male chickens, the effect of tannin supplementation (0, 250, 500 and 1000 mg/kg) on performance, microbial status of chickens small intestine and colon of 28- and 41-d-old chickens, as well as histological changes of jejunum walls at 41 d and carcase quality were determined. 2. Application of 250 or 500 mg of sweet chestnut tannin per kg of feed had an insignificant effect on body weight and feed conversion of 41-d-old chickens (30 and 26%) in comparison to control birds. The highest tannin supplement (1000 mg/kg) reduced final body weight. 3. No effects of tannin supplementation on feed conversion and carcase quality were found. 4. Addition of tannin increased dry matter content of the litter by 88 (Group II) and 77% (Group III) when compared to control. 5. Higher doses of tannins significantly reduced the number of E. coli and coliform bacteria in small intestine of 28-d-old chickens; in other microorganisms great variability of microbial populations in small intestine and colon were observed. 6. The histologies of jejunal walls in chickens of control, II (250 mg/kg) and III (500 mg/kg) groups were similar. The structure was characteristic of correctly developed and functioning tissues and the villi were formed correctly. Tannin applied at the highest dose (1000 mg) slowed down the proliferation rate in the mother-cell zone. Single cells and enterocyte complexes showed the features characteristic of degradation processes. These unfavourable symptoms indicated some disturbances in intestinal wall morphology and function.


Psip1/p52 regulates posterior Hoxa genes through activation of lncRNA Hottip.
Long noncoding RNAs (lncRNAs) have been implicated in various biological functions including the regulation of gene expression, however, the functionality of lncRNAs is not clearly understood and conflicting conclusions have often been reached when comparing different methods to investigate them. Moreover, little is known about the upstream regulation of lncRNAs. Here we show that the short isoform (p52) of a transcriptional co-activator-PC4 and SF2 interacting protein (Psip1), which is known to be involved in linking transcription to RNA processing, specifically regulates the expression of the lncRNA Hottip-located at the 5' end of the Hoxa locus. Using both knockdown and knockout approaches we show that Hottip expression is required for activation of the 5' Hoxa genes (Hoxa13 and Hoxa10/11) and for retaining Mll1 at the 5' end of Hoxa. Moreover, we demonstrate that artificially inducing Hottip expression is sufficient to activate the 5' Hoxa genes and that Hottip RNA binds to the 5' end of Hoxa. By engineering premature transcription termination, we show that it is the Hottip lncRNA molecule itself, not just Hottip transcription that is required to maintains active expression of posterior Hox genes. Our data show a direct role for a lncRNA molecule in regulating the expression of developmentally-regulated mRNA genes in cis.


Paper-based electrodes modified with cobalt phthalocyanine colloid for the determination of hydrogen peroxide and glucose.
Cobalt(ii) phthalocyanine (CoPc) was suspended in aqueous medium and the colloidal system was used as catalyst for the electrochemical determination of hydrogen peroxide on paper-based electrodes modified with carbon nanomaterials. H2O2 was oxidised at 0.275 V vs. Ag pseudoreference electrode. This system was adapted to develop a glucose sensor with glucose oxidase immobilized on the cellulose electrode. CoPc suspended nanoparticles acted as nanoenzyme mimicking peroxidase activity and were combined with different carbon nanomaterials to form hybrids with optimised catalytic performance. GO-CoPc paper-based electrodes yielded the best results with a linear range of ∼12 μM to 49 mM for H2O2 and 0.1 mM to 1 mM for glucose. Glucose was determined in physiological serum and juice samples with recoveries of 93.3 and 94.2% respectively. CoPc could replace HRP for the catalytic sensing of H2O2, without the need to be dissolved. This material can be used in situ in a simple protocol with other nanomaterials for electrode modification. The sensor described has the advantage of easy preparation, using the catalyst in colloidal form, long term stability, and versatility to be adapted to other low cost and disposable enzymatic systems.


Trk B signalling controls LTP but not LTD expression in the developing rat visual cortex.
Neurotrophins have been suggested to act as liaison molecules between activity-dependent synaptic plasticity and the establishment of patterns of synaptic connectivity during postnatal developmental in different brain areas, including the visual cortex. In particular, recent studies have shown that Trk B ligands are involved in the formation of the ocular dominance columns during postnatal development. Here, we examined the contribution of endogenous Trk B activation to the regulation of different forms of synaptic plasticity including long-term potentiation (LTP), long-term depression (LTD) and LTP after LTD in the developing visual cortex. Rat cortical slices were incubated with a soluble form of Trk B receptor (TrkB IgG) preventing Trk B activation by endogenous ligands. LTP expression was also studied at P23 (postnatal), when the expression of brain-derived neurotrophic factor (BDNF) reaches a peak and the LTP expression is normally downregulated. The present results demonstrate that Trk B activation is required for the long-term maintenance, > 30 min, of both LTP and LTP after LTD at P17. At P23, a higher concentration of TrkB IgG was necessary to impair LTP. In contrast, neither amplitude nor duration of LTD were affected by Trk B ligands blockade. Taken together, these results indicate that endogenous Trk B ligands are necessary for the expression of LTP but not LTD at a critical time during postnatal cortical development.


Cloning of the cDNA encoding the urotensin II precursor in frog and human reveals intense expression of the urotensin II gene in motoneurons of the spinal cord.
Urotensin II (UII) is a cyclic peptide initially isolated from the caudal neurosecretory system of teleost fish. Subsequently, UII has been characterized from a frog brain extract, indicating that a gene encoding a UII precursor is also present in the genome of a tetrapod. Here, we report the characterization of the cDNAs encoding frog and human UII precursors and the localization of the corresponding mRNAs. In both frog and human, the UII sequence is located at the C-terminal position of the precursor. Human UII is composed of only 11 amino acid residues, while fish and frog UII possess 12 and 13 amino acid residues, respectively. The cyclic region of UII, which is responsible for the biological activity of the peptide, has been fully conserved from fish to human. Northern blot and dot blot analysis revealed that UII precursor mRNAs are found predominantly in the frog and human spinal cord. In situ hybridization studies showed that the UII precursor gene is actively expressed in motoneurons. The present study demonstrates that UII, which has long been regarded as a peptide exclusively produced by the urophysis of teleost fish, is actually present in the brain of amphibians and mammals. The fact that evolutionary pressure has acted to conserve fully the biologically active sequence of UII suggests that the peptide may exert important physiological functions in humans.


Interaction of deoxycholate with the sodium channel of squid axon membranes.
Deoxycholate can react with sodium channels with a high potency. The apparent dissociation constant for the saturable binding reaction is 2 microM at 8 degrees C, and the heat of reaction is approximately -7 kcal/mol. Four independent test with Na-free media, K-free media, tetrodotoxin, and pancuronium unequivocally indicate that it is the sodium channel that is affected by deoxycholate. Upon depolarization of the membrane, the drug modified channel exhibits a slowly activating and noninactivating sodium conductance. The kinetic pattern of the modified channel was studied by increasing deoxycholate concentration, lowering the temperature, chemical elimination of sodium inactivation, or conditioning depolarization. The slow activation of the modified channel can be represented by a single exponential function with the time constant of 1--5 ms. The modified channel is inactivated only partially with a time constant of 1 S. The reversal potential is unchanged by the drug. Observations in tail currents and the voltage dependence of activation suggest that the activation gate is actually unaffected. The apparently slow activation may reflect an interaction betweem deoxycholate and the sodium channel in resting state.


ID1 contributes to cell growth invasion and migration in salivary adenoid cystic carcinoma.
Previous studies have reported that inhibitor of DNA binding 1 (ID1) exerts an oncogenic role in a number of tumors. In the present study, the role of ID1 in the growth, invasion and migration of salivary adenoid cystic carcinoma (SACC) cells was investigated. ID1 expression in clinical SACC samples was compared with that in normal salivary tissues using immunohistochemical staining, and the correlation between ID1 expression and clinical pathological characteristics was then determined. Subsequently, ID1 was overexpressed or silenced to investigate the effects of ID1 expression on SACC cell proliferation, invasion and migration. In addition, the gene expression levels of known ID1 target genes, including S100A9, CDKN2A and matrix metalloproteinase 1 (MMP1) was measured using reverse transcription‑quantitative polymerase chain reaction to elucidate the potential mechanisms of ID1 in SACC. The results of the present study indicated that the protein expression levels of ID1 were significantly increased in the SACC tissues compared with that in the normal salivary tissues (P<0.001), and a positive correlation between ID1 expression and tumor stage (P=0.001), tumor invasion (P=0.002) and metastasis (P=0.019) in SACC was observed. Knockdown of ID1 in SACC cells significantly inhibited cell growth, invasion and migration (all P<0.01), whereas overexpression of ID1 promoted cell proliferation, invasion and migration (all P<0.01). The gene expression level of MMP1 was significantly reduced following ID1 knockdown in SACC‑83 cells when compared with negative controls (P<0.05), whereas S100A9 and CDKN2A expression levels were significantly upregulated (both P<0.05). The results suggest that ID1 may regulate the growth, invasion and migration of SACC cells, and that MMP1, S100A9 and CDKN2A may serve as target genes of ID1 and mediate the effects of ID1 in SACC cells. Therefore, ID1 may present a potential target gene for the treatment of patients with SACC to inhibit cancer cell growth and metastasis.


The Ess1 prolyl isomerase is dispensable for growth but required for virulence in Cryptococcus neoformans.
Cryptococcus neoformans is an important human fungal pathogen that also serves as a model for studies of fungal pathogenesis. C. neoformans contains several genes encoding peptidyl-prolyl cis/trans isomerases (PPIases), enzymes that catalyse changes in the folding and conformation of target proteins. Three distinct classes of PPIases have been identified: cyclophilins, FK506-binding proteins (FKBPs) and parvulins. This paper reports the cloning and characterization of ESS1, which is believed to be the first (and probably only) parvulin-class PPIase in C. neoformans. It is shown that ESS1 from C. neoformans is structurally and functionally homologous to ESS1 from Saccharomyces cerevisiae, which encodes an essential PPIase that interacts with RNA polymerase II and plays a role in transcription. In C. neoformans, ESS1 was found to be dispensable for growth, haploid fruiting and capsule formation. However, ESS1 was required for virulence in a murine model of cryptococcosis. Loss of virulence might have been due to the defects in melanin and urease production observed in ess1 mutants, or to defects in transcription of as-yet-unidentified virulence genes. The fact that Ess1 is not essential in C. neoformans suggests that, in this organism, some of its functions might be subsumed by other prolyl isomerases, in particular, cyclophilins Cpa1 or Cpa2. This is supported by the finding that ess1 mutants were hypersensitive to cyclosporin A. C. neoformans might therefore be a useful organism in which to investigate crosstalk among different families of prolyl isomerases.


Considerations for the use of virally delivered genetic tools for in-vivo circuit analysis and behavior in mutant mice: a practical guide to optogenetics.
Optogenetics was the method of the year in 2010 according to Nature Neuroscience. Since then, this method has become widespread, the use of virally delivered genetic tools has extended to other fields such as pharmacogenetics, and optogenetic techniques have become frequently applied in genetically manipulated animals for in-vivo circuit analysis and behavioral studies. However, several issues should be taken into consideration when planning such experiments. We aimed to summarize the critical points concerning optogenetic manipulation of a specific brain area in mutant mice. First, the appropriate vector should be chosen to allow optimal optogenetic manipulation. Adeno-associated viral vectors are the most common carriers with different available serotypes. Light-sensitive channels are available in many forms, and the expression of the delivered genetic material can be influenced in many ways. Second, selecting the adequate stimulation protocol is also essential. The pattern, intensity, and timing could be determinative parameters. Third, the mutant strain might have a phenotype that influences the observed behavior. In conclusion, detailed preliminary experiments and numerous control groups are required to choose the best vector and stimulation protocol and to ensure that the mutant animals do not have a specific phenotype that can influence the examined behavior.


TRPC6 channels and their binding partners in podocytes: role in glomerular filtration and pathophysiology.
Loss or dysfunction of podocytes is a major cause of glomerular kidney disease. Several genetic forms of glomerular disease are caused by mutations in genes that encode structural elements of the slit diaphragm or the underlying cytoskeleton of podocyte foot processes. The recent discovery that gain-of-function mutations in Ca(2+)-permeable canonical transient receptor potential-6 channels (TRPC6) underlie a subset of familial forms of focal segmental glomerulosclerosis (FSGS) has focused attention on the basic cellular physiology of podocytes. Several recent studies have examined the role of Ca(2+) dynamics in normal podocyte function and their possible contributions to glomerular disease. This review summarizes the properties of TRPC6 and related channels, focusing on their permeation and gating properties, the nature of mutations associated with familial FSGS, and the role of TRPC channels in podocyte cell biology as well as in glomerular pathophysiology. TRPC6 interacts with several proteins in podocytes, including essential slit diaphragm proteins and mechanosensitive large-conductance Ca(2+)-activated K(+) channels. The signaling dynamics controlling ion channel function and localization in podocytes appear to be quite complex.


Preoperative predictors of late postoperative outcome among patients with nonischemic mitral regurgitation with 'high risk' descriptors and comparison with unoperated patients.
Among patients with chronic nonischemic mitral regurgitation (MR), high short-term mortality risk can be identified by left (LV) and/or right ventricular (RV) ejection fraction (EF) criteria (LVEF </=45% and/or RVEF </=30%). Mitral valve replacement or repair (MVR) significantly improves outcome in this subgroup, but predictors of late postoperative survival are not known, and the benefit of MVR has not been defined in patients matched for severity of LV and RV dysfunction. Therefore, prospective assessment of 14 consecutive high risk MR patients was performed before MVR and during 9 years (average) postoperatively to define echocardiographic and radionuclide angiographic predictors of survival; survival also was evaluated in a contemporaneous series of 9 high risk unoperated MR patients, and in subgroups of operated and unoperated patients matched for EF. Of 14 MVR patients, 4 died (3 cardiac: 1 sudden, 2 congestive heart failure). Only preoperative RVEF </=20% significantly predicted postoperative deaths (rest p = 0.032; exercise p = 0.05). Of 9 unoperated patients, 8 died. Mortality risk of unoperated patients remained higher than that of MVR patients when groups were matched for preoperative LVEF (p = 0.0001). Among patients with RVEF >20%, MVR significantly improved survival versus medical treatment (rest: p < 0.0001, exercise: p = 0.0003). In high risk MR patients, MVR improves survival; preoperative RV performance can define subgroups with different long-term postoperative survival.


Preventing renal complications in type 2 diabetes: results of the diabetics exposed to telmisartan and enalapril trial.
Patients with type 2 diabetes are prone to hypertension and persistent protein leakage from the kidney (microalbuminuria or macroalbuminuria). A progressive decline in renal function can lead to overt diabetic nephropathy and ESRD. The likelihood of cardiovascular disease also is increased. Control of hypertension is paramount to prevent these life-threatening complications. Agents that target the renin-angiotensin system--angiotensin-converting enzyme inhibitors and angiotensin II receptor blockers--have been shown to be renoprotective. The groundbreaking Diabetics Exposed to Telmisartan And enalaprIL (DETAIL) trial was designed to address the absence of comparative data on the long-term effects of an angiotensin II receptor blocker versus an angiotensin-converting enzyme inhibitor on renoprotection and survival in 250 patients with hypertension and early type 2 diabetic nephropathy. The primary purpose of the 5-yr double-blind, double-dummy, randomized study was to establish whether 40 to 80 mg of telmisartan conferred similar (i.e., noninferior) renoprotection to 10 to 20 mg of enalapril as determined by the change from baseline in GFR, measured by the plasma clearance of iohexol. Secondary end points included the emergence of ESRD and all-cause mortality. Telmisartan was not inferior to enalapril in reducing the decline in GFR: Mean annual declines in GFR were 3.7 and 3.3 ml/min per 1.73 m(2) with telmisartan and enalapril, respectively. During the 5-yr study period, no patient developed a serum creatinine >200 micromol/L, and none required dialysis. There were only six deaths in each treatment group during the study, with half being due to cardiovascular events.


DUSP1 is a novel target for enhancing pancreatic cancer cell sensitivity to gemcitabine.
Pancreatic ductal adenocarcinoma (PDAC) is a deadly cancer with a poor prognosis that is characterized by excessive mitogenic pathway activation and marked chemoresistance to a broad spectrum of chemotherapeutic drugs. Dual specificity protein phosphatase 1 (DUSP1) is a key negative regulator of mitogen activated protein kinases (MAPKs). Yet, DUSP1 is overexpressed in pancreatic cancer cells (PCCs) in PDAC where it paradoxically enhances colony formation in soft agar and promotes in vivo tumorigenicity. However, it is not known whether DUSP1 overexpression contributes to PDAC chemoresistance. Using BxPC3 and COLO-357 human PCCs, we show that gemcitabine activates c-JUN N-terminal kinase (JNK) and p38 mitogen activated protein kinase (p38 MAPK), key kinases in two major stress-activated signaling pathways. Gemcitabine-induced JNK and p38 MAPK activation mediates increased apoptosis, but also transcriptionally upregulates DUSP1, as evidenced by increased DUSP1 mRNA levels and RNA polymerase II loading at DUSP1 gene body. Conversely, shRNA-mediated inhibition of DUSP1 enhances JNK and p38 MAPK activation and gemcitabine chemosensitivity. Using doxycycline-inducible knockdown of DUSP1 in established orthotopic pancreatic tumors, we found that combining gemcitabine with DUSP1 inhibition improves animal survival, attenuates angiogenesis, and enhances apoptotic cell death, as compared with gemcitabine alone. Taken together, these results suggest that gemcitabine-mediated upregulation of DUSP1 contributes to a negative feedback loop that attenuates its beneficial actions on stress pathways and apoptosis, raising the possibility that targeting DUSP1 in PDAC may have the advantage of enhancing gemcitabine chemosensitivity while suppressing angiogenesis.


Heavy and light chains of a mouse monoclonal autoantibody express the same idiotype.
We report on the detection of the same idiotype (Id62) on the heavy and light chains of a mouse monoclonal autoantibody to thyroglobulin. This result was obtained by a simple, sensitive, and reproducible technique, namely immunoblot on nitrocellulose paper of electrophoretically separated heavy and light chains by using a panel of 125I-labeled anti-idiotype probes. These included polyclonal heterologous and syngeneic affinity-purified antibodies and a syngeneic monoclonal antibody. The immunoblot findings were confirmed through the solid-phase radioimmunoassay binding of the anti-idiotype probes to isolated H and L polypeptide chains prepared by conventional chromatography methods. Because anti-idiotype probes define a putative regulatory idiotype of the BALB/c autoimmune response to thyroglobulin, it is suggested that the simultaneous presence of the same idiotype on H and L chains may confer to antibody regulatory properties within the idiotypic network. The significance of the existence of the same idiotype on H and L chains and the implication of this finding for the regulation of the immune response to self antigens are discussed.


Identifying the snake: First scoping review on practices of communities and healthcare providers confronted with snakebite across the world.
Snakebite envenoming is a major global health problem that kills or disables half a million people in the world's poorest countries. Biting snake identification is key to understanding snakebite eco-epidemiology and optimizing its clinical management. The role of snakebite victims and healthcare providers in biting snake identification has not been studied globally. This scoping review aims to identify and characterize the practices in biting snake identification across the globe. Epidemiological studies of snakebite in humans that provide information on biting snake identification were systematically searched in Web of Science and Pubmed from inception to 2nd February 2019. This search was further extended by snowball search, hand searching literature reviews, and using Google Scholar. Two independent reviewers screened publications and charted the data. We analysed 150 publications reporting 33,827 snakebite cases across 35 countries. On average 70% of victims/bystanders spotted the snake responsible for the bite and 38% captured/killed it and brought it to the healthcare facility. This practice occurred in 30 countries with both fast-moving, active-foraging as well as more secretive snake species. Methods for identifying biting snakes included snake body examination, victim/bystander biting snake description, interpretation of clinical features, and laboratory tests. In nine publications, a picture of the biting snake was taken and examined by snake experts. Snakes were identified at the species/genus level in only 18,065/33,827 (53%) snakebite cases. 106 misidentifications led to inadequate victim management. The 8,885 biting snakes captured and identified were from 149 species including 71 (48%) non-venomous species. Snakebite victims and healthcare providers can play a central role in biting snake identification and novel approaches (e.g. photographing the snake, crowdsourcing) could help increase biting snake taxonomy collection to better understand snake ecology and snakebite epidemiology and ultimately improve snakebite management.


Lignin Induces ES Cells to Differentiate into Neuroectodermal Cells through Mediation of the Wnt Signaling Pathway.
Embryonic stem cells (ES cells) are characterized by their pluripotency and infinite proliferation potential. Ever since ES cells were first established in 1981, there have been a growing number of studies aimed at clinical applications of ES cells. In recent years, various types of differentiation inducement systems using ES cells have been established. Further studies have been conducted to utilize differentiation inducement systems in the field of regenerative medicine. For cellular treatments using stem cells including ES cells, differentiation induction should be performed in a sufficient manner to obtain the intended cell lineages. Lignin is a high-molecular amorphous material that forms plants together with cellulose and hemicelluloses, in which phenylpropane fundamental units are complexly condensed. Lignin derivatives have been shown to have several bioactive functions. In spite of these findings, few studies have focused on the effects of lignin on stem cells. Our study aimed to develop a novel technology using lignin to effectively induce ES cells to differentiate into neuroectodermal cells including ocular cells and neural cells. Since lignin can be produced at a relatively low cost in large volumes, its utilization is expected for more convenient differentiation induction technologies and in the field of regenerative medicine in the future.


Early socioeconomic adversity and young adult physical illness: the role of body mass index and depressive symptoms.
The present study investigated the psychophysiological inter- and intra-individual processes that mediate the linkage between childhood and/or adolescent socioeconomic adversities and adult health outcomes. Specifically, the proposed model examined the roles of youth depressive symptoms and body mass index (BMI) trajectories as mediators that explain the link between early adversity and young adults' general health and physical illnesses after controlling for gender, race or ethnicity, and earlier general health reports. Using a nationally representative sample of 12,424 from National Longitudinal Study of Adolescent Health (Add Health), this study used growth curve modeling to consider both the severity (initial level) and the change over time (deterioration or elevation) as psychophysiological mediators, thereby acknowledging multiple facets of depressive symptoms and BMI trajectories as psychophysiological mediators of early adversity to adult health. Results provide evidence for (1) the influence of early childhood and early adolescent cumulative socioeconomic adversity on both the initial levels and changes over time of depressive symptoms and BMI and (2) the independent influences depressive symptoms and BMI trajectories on the general health and the physical illnesses of young adults. These findings contribute valuable knowledge to existing research by elucidating how early adversity exerts an enduring long-term influence on physical health problems in young adulthood; furthermore, this information suggests that effective intervention and prevention programs should incorporate multiple facets (severity and change over time) of multiple mechanisms (psychological and physiological).


Development and Implementation of Advanced Fitting Methods for the Calculation of Accurate Molecular Structures.
The determination of accurate equilibrium molecular structures plays a fundamental role for understanding many physical-chemical properties of molecules, ranging from the precise evaluation of the electronic structure to the analysis of dynamical and environmental effects in tuning their overall behavior. For this purpose the so-called semiexperimental approach, based on a nonlinear least-squares fit of the moments of inertia associated with a set of available isotopologues, allows one to obtain very accurate results, without the unfavorable computational cost characterizing high-level quantum chemical methods. In the present work the MSR (Molecular Structure Refinement) software for the determination of equilibrium structures by means of the semiexperimental approach is presented, and its implementation is discussed in some detail. The software, which is interfaced with a powerful graphical user interface, includes different optimization algorithms, an extended error analysis, and a number of advanced features, the most remarkable ones concerning the choice of internal coordinates and the method of predicate observations. In particular, a new black-box scheme for defining automatically a suitable set of nonredundant internal coordinates of A1 symmetry in place of the customary Z-matrix has been designed and tested. Finally, the implementation of the method of the predicate observations is discussed and validated for a set of test molecules. As an original application, the method is employed for the determination of the semiexperimental structure for the most stable conformer of glycine.


Repeatability and Reproducibility of Total Retinal Blood Flow Measurements Using Bi-Directional Doppler OCT.
To investigate the repeatability and reproducibility of total retinal blood flow measurements using a custom-built dual-beam bidirectional Doppler optical coherence tomography (OCT) system in healthy subjects. Repeatability and reproducibility were analyzed in 10 and 34 healthy subjects, respectively. For repeatability, measurements were taken twice within 30 minutes, for reproducibility, twice within two to five weeks. Two analysis approaches were compared for calculation of absolute blood velocities: a previously published approach resulting in values for total arterial (QA,abs) and total venous blood flow (QV,abs) and a novel approach taking into account that there is a fixed relation between the phase shift in the two OCT channels (QA,new, QV,new). Repeatability and reproducibility were quantified using intraclass correlation coefficients (ICCs). For QA,abs and QV,abs, ICC values between 0.78 and 0.84 were obtained. QA,new and QV,new values revealed better repeatability and reproducibility as compared to the convential appoach. Repeatability ICCs for QA,new and QV,new were between 0.91 and 0.93, and reproducibility ICCs were between 0.87 and 0.91 indicating excellent reproducibility. Good agreement was observed between total retinal blood flow values as measured from retinal arteries and retinal veins. Measurement of total retinal blood flow using dual-beam Doppler OCT shows excellent reproducibility, which can further be improved by using a novel algorithm for calculating blood velocities in retinal vessels. Our data indicate that dual-beam Doppler OCT can be used for longitudinal studies. Hence, quantitative retinal blood flow may be established as a biomarker for progression vascular eye diseases.


Monoclonal antibodies specific for the amastigote stage of Leishmania pifanoi. I. Characterization of antigens associated with stage- and species-specific determinants.
Eight mAb were produced against membrane-enriched preparations of Leishmania pifanoi amastigotes either grown in axenic culture (P-1 through P-6) or isolated from macrophage cell culture (P-7 and P-8). Two mAb produced against promastigote membranes (P-9 and P-10) were found to be specific against this stage. Antibodies P-1 through P-8 on analysis by radioimmune binding only reacted with determinants present on amastigotes. mAb P-2, P-4, and P-8 also reacted with Leishmania amazonensis amastigotes but not promastigotes. No cross-reactions were found on any other species of Leishmania or with membranes of Trypanosoma cruzi epimastigotes or amastigotes. An indirect immunofluorescence assay using mAb P-1 through P-8 confirmed the stage specificity and binding to L. pifanoi axenically grown amastigotes, amastigotes within infected hamster tissue, and amastigotes within J774.1 macrophages. When Western blot analysis of amastigote membranes was conducted, one distinct group of molecules associated with L. pifanoi-specific determinants was identified. mAb P-1, P-3, P-5, P-7, and P-8 bound to molecules Mr 43 and 34 kDa. Promastigote-specific mAb P-9 recognized a diffuse pattern from 88 to greater than 200 kDa, and mAb P-10 localized a second class of proteins with Mr53 kDa. On immunoprecipitation of solubilized [35S]methionine-labeled amastigotes, mAb P-2 recognized a doublet of Mr 35 and 33 kDa and another doublet at Mr 17.5 and 13.5 kDa. mAb P-4 and P-7 each precipitated a band at Mr 34 kDa. These studies indicate that antigenically the axenically cultured amastigote is closely related to macrophage-derived amastigote. These mAb and/or purified protein Ag may be useful in studying stage differentiation, monitoring transformation, and for further taxonomic, epidemiologic, and immunologic studies of New World leishmaniasis.


Primary musculoskeletal sarcoidosis.
Sarcoidosis is a well-described clinical entity with several types of presentation among different organ systems. However, primary musculoskeletal manifestation is a rare and seldom-described manifestation. A retrospective review of patients seen at our institution from 1999 to 2009 was performed to identify patients with such presentation. Eleven patients with musculoskeletal sarcoidosis were identified. Seven patients presented with solitary lesions, while 4 presented with multiple foci of disease. Histologically, all lesions revealed granulomatous inflammation consistent with sarcoidosis. Cultures and gram stain were negative in all patients. One patient was found to have evidence of pulmonary sarcoidosis after diagnosis of musculoskeletal sarcoidosis. Eight patients had primarily soft tissue involvement with visible masses, while 3 patients had primarily osseous involvement with no mass. Pain was reported by 9 of 11 patients. On magnetic resonance imaging, all lesions were dark on T1, bright on T2, and poorly marginated. Magnetic resonance imaging was noted to be more accurate in revealing the pathology than radiographic imaging. Four patients underwent complete excision and 1 received a steroid injection. The remaining patients required no further treatment after diagnosis. Musculoskeletal sarcoidosis is rare and can present as single or multiple lesions. Definitive diagnosis is made by biopsy. The decision to operate should be based on symptoms, issues with cosmesis, impending fractures, growth, and/or involvement of neuro-vascular structures. Patients should be evaluated for pulmonary involvement.


Per-oral endoscopic myotomy (POEM): a new endoscopic treatment for achalasia.
Per-oral endoscopic myotomy (POEM) is a new minimally invasive technique to treat achalasia. We performed a review of the literature of POEM with a special focus on technical details and the results obtained with this technique in patients with achalasia and other esophageal motility disorders. Thousands of POEM procedures have been performed worldwide since its introduction in 2008. The procedure is based on the creation of a mucosal entry point in the proximal esophagus to reach the cardia through a submucosal tunnel and then perform a myotomy of the muscular layers of the cardia, esophagogastric junction and distal esophagus, as performed in a Heller myotomy. The clinical remission rate ranges from 82 to 100%. Although no randomized studies exist and available data are from single-center studies, no differences have been found between laparoscopic Heller myotomy (LHM) and POEM in terms of perioperative outcomes, short-term outcomes (12 months) and long-term outcomes (up to three years). Procedure time and length of hospital stay were lower for POEM. Post-POEM reflux is a concern, and controversial data have been reported compared to LHM. The technique is safe, with no reported deaths related to the procedure and an adverse event rate comparable to surgery. Potential complications include bleeding, perforation, aspiration and insufflation-related adverse events. Thus, this is a complex technique that needs specific training even in expert hands. The indication for this procedure is widening and other motor hypercontractil esophageal disorders have been treated by POEM with promising results. POEM can be performed in complicated situations such as in pediatric patients, sigmoid achalasia or after failure of previous treatments. POEM is an effective treatment for achalasia and is a promising tool for other motor esophageal disorders. It is a safe procedure but, due to its technical difficulty and possible associated complications, the procedure should be performed in referral centers by trained endoscopists.