Morphological aspects of ciliary motility.
In Elliptio complanatus lateral cilia, two distinct patterns of filament termination can be discerned. In one case, all nine filaments are present and all are single; in the second, at least one filament is missing but doublets are still present. These probably represent different configurations within one cilium in different stroke positions; to get from one to the other, some peripheral filaments must move with respect to others. The data are consistent with the hypothesis that the filaments themselves do not change length, but rather slide past one another to accommodate increasing curvature. The bent regions of the cilium are in the form of circular arcs. In a few cases, apparent displacement of filaments at the tip (Deltal) can be shown to be accounted for if we assume that all differences are generated within these arcs. The displacement per degree of bend is 35 A. Regions of bent arc are initially confined to the base of the cilium but move up the shaft as straight regions appear below them. From the relationship between arc length and radius of curvature, a shaft length that is the unit that initially bends and slides may be defined. Quantal displacements of the length of one 14S dynein may perhaps occur at sites between filaments at opposite sides of such a unit as sliding occurs.


Plexiform neurofibroma of the submandibular gland in patient with von Recklinghausen's disease.
Plexiform neurofibroma of the submandibular gland is an extremely rare tumor. Herein, we report a case of plexiform neurofibroma in a patient with a von Recklinghausen's disease (NF-1) who presented with a submandibular mass mimicking a submandibular gland tumor. Complete surgical excision provides the best treatment and final diagnosis. A neurofibroma should be considered in the differential diagnosis for submandibular mass.


Health Inequality between Migrant and Non-Migrant Workers in an Industrial Zone of Vietnam.
Vietnam has experienced massive internal migration waves from rural to industrialized zones. However, little efforts have been made to understand differences in health conditions and health-related quality of life (HRQOL) between local and migrant industrial workers. This study aimed to examine the inequality in health status and HRQOL between these workers. We conducted a cross-sectional study of 289 Vietnamese workers at three industrial areas in Hanoi and Bac Ninh. Self-reported health status and HRQOL were measured using the EuroQOL-5 dimensions-5 levels (EQ-5D-5L) instrument. Sociodemographic, working, and environmental factors were also investigated. Overall, the mean EQ-5D index was 0.74 (SD = 0.21) and the average number of health problems in the last 12 months in our sample was 1.91 (SD = 1.63) problems. Migrant people had a lower EQ-5D index (β = -0.08, p < 0.01) and more health problems (β = 0.20, p < 0.05) compared to local workers. Those being male, working in the same posture more than 60 min, and exposed to more hazards at work were correlated with a lower EQ-5D index and higher number of health problems. The results highlighted inequalities in health status and HRQOL between migrant and local workers. Reinforcing regular health check-ups, ensuring sufficient protective equipment and working conditions may help improve the health outcomes of the workers.


Complex muscle vibration patterns to induce gait-like lower-limb movements: proof of concept.
Muscle vibrations can induce motor responses and illusions of complex movements. However, inducing gait-like cyclical movements and illusions requires the application of multiple fast alternating vibrations to lower-limb muscles. The objectives were (1) to test the feasibility of delivering complex vibrations in a time-organized manner and (2) to illustrate the possibility of inducing alternate gait-in-place-like movements using these vibrations. Patterns of vibration, produced by 12 vibrators applied bilaterally on the flexor and extensor muscle groups of the lower limbs, were based on normal gait kinematics. We tested 1 s and 2 s cycle patterns of vibration. Vibrator responses were assessed using auto- and crosscorrelations and frequency analyses based on accelerometry measurements, and compared between patterns. High auto- (>0.8) and crosscorrelation (>0.6) coefficients demonstrated a good response by the vibrators to the control signal. Vibrations induced cyclical, low-amplitude stepping-in-place movements that mimicked alternate walking movements with both legs, with 1 s and 2 s cycle durations, in one nondisabled participant and one participant with American Spinal Injury Association Impairment Scale B spinal cord injury standing, relaxed, with body-weight support. Electromechanical vibrators can deliver complex cyclical vibrations and trigger gait-like lower-limb movements. These results warrant the application of these vibration patterns on individuals with sensorimotor impairments to test their potential in gait rehabilitation.


The outcome of pediatric patients undergoing congenital cardiac surgery under pulsatile cardiopulmonary bypass in different frequencies.
To investigate the influence and possible pathophysiological mechanism of pulsatile cardiopulmonary bypass (CPB) in various frequencies in pediatric patients undergoing congenital cardiac surgery. Clinical data and hemodynamic parameters were collected in 80 patients who underwent congenital cardiac surgeries and were perfused in different settings: pulsatile perfusion (PP) in frequencies of 30 beats/min, PP 60 beats/min, PP 100 beats/min and non-pulsatile perfusion (NP). Serum proteins, plasma-free hemoglobin (PFH), endothelin-1 (ET-1) and nitric oxide (NO) were collected to study possible pathophysiological changes, possible hematological injury and oxidative status under different perfusing conditions. Patients in all groups had similar baseline characteristics, aortic cross-clamping time and CPB duration. More effective pulse gradient (PG), energy-equivalent pressure (EEP) and surplus hemodynamic energy (SHE) were observed in pulsatility with lower frequency setting, under which more patients achieved physiologically normal mean arterial pressure (MAP), without the support of inotropic agents during bypass. Significant between-group differences of serum proteins and PFH were absent the whole time during and after bypass, while a relatively lower percentage of perioperative requirement of diuretics was observed in the low frequency pulsatile group. A better performance to oxidative stress was seen in the low frequency group with higher levels of NO and lower concentration of ET-1, and both intergroup differences were found (P<0.01). Satisfactory clinical outcome was obtained on post procedure course in all groups. Pulsatile perfusion with low frequency setting in pediatric patients undergoing congenital cardiac surgery showed better hemodynamic profiles, potential protective effects on vital organs, better oxidative status and satisfactory clinical outcome.


G-Protein binding domains of the angiotensin II AT1A receptors mapped with synthetic peptides selected from the receptor sequence.
The vascular angiotensin II type 1 receptor (AT1AR) is a member of the G-protein-coupled receptor superfamily. We mapped the G-protein binding domains of the AT1AR using synthetic peptides selected from the receptor sequence, which interfere with AT1AR-G-protein coupling. Membrane GTPase activity was used as a measure of the functional coupling in rat vascular smooth muscle cells. Peptides corresponding to the N-terminal region of the second intracellular loop (residues 125-137), the N-terminal region of the third intracellular loop (217-227) and the juxtamembranous region of the C-terminal tail (304-316) inhibited angiotensin II-induced GTPase activation by 30%, 30%, and 70%, respectively. The latter two domains (217-227 and 304-316) are predicted to form amphiphilic alpha-helices. Only the peptide representing residues 217-227 stimulated basal activity (45%). No synthetic peptide had a significant effect on either the number or the affinity of the AT1AR binding. These observations indicate that domains of the second and third regions and the cytoplasmic tail of the AT1AR interact with G-proteins, and that multiple contacts with these receptor domains may be important for binding and activation of the G-proteins.


The Timothy syndrome mutation differentially affects voltage- and calcium-dependent inactivation of CaV1.2 L-type calcium channels.
Calcium entry into excitable cells is an important physiological signal, supported by and highly sensitive to the activity of voltage-gated Ca2+ channels. After membrane depolarization, Ca2+ channels first open but then undergo various forms of negative feedback regulation including voltage- and calcium-dependent inactivation (VDI and CDI, respectively). Inactivation of Ca2+ channel activity is perturbed in a rare yet devastating disorder known as Timothy syndrome (TS), whose features include autism or autism spectrum disorder along with severe cardiac arrhythmia and developmental abnormalities. Most cases of TS arise from a sporadic single nucleotide change that generates a mutation (G406R) in the pore-forming subunit of the L-type Ca2+ channel Ca(V)1.2. We found that the TS mutation powerfully and selectively slows VDI while sparing or possibly speeding the kinetics of CDI. The deceleration of VDI was observed when the L-type channels were expressed with beta1 subunits prominent in brain, as well as beta2 subunits of importance for the heart. Dissociation of VDI and CDI was further substantiated by measurements of Ca2+ channel gating currents and by analysis of another channel mutation (I1624A) that hastens VDI, acting upstream of the step involving Gly406. As highlighted by the TS mutation, CDI does not proceed to completeness but levels off at approximately 50%, consistent with a change in gating modes and not an absorbing inactivation process. Thus, the TS mutation offers a unique perspective on mechanisms of inactivation as well as a promising starting point for exploring the underlying pathophysiology of autism.


Impact of a formal breastfeeding education program.
To investigate whether implementation of a formal breastfeeding education program including a breastfeeding protocol, a resource guide, and educational presentations would have an impact on knowledge, comfort level, and attitudes toward breastfeeding among healthcare providers, and on amount of exclusive breastfeeding at our hospital. Quasi-experimental study that included a survey to obtain a preintervention baseline measurement and a postintervention measurement 6 months after the implementation of the formal breastfeeding education program. The sample was obstetricians, pediatricians, and nurses who interact with breastfeeding mothers at our hospital. Additionally, we surveyed breastfeeding mothers who gave birth at our hospital to determine the impact on the nurse observation of breastfeeding, exclusivity of breastfeeding, and nighttime feedings. Healthcare providers showed increased levels of knowledge and comfort dealing with breastfeeding issues after the education program. Knowledge scores were significantly improved (pre- vs. postimplementation scores were 20.2 vs. 22.2, p < .05). Comfort level scores were also significantly improved (pre- vs. postimplementation scores were 20.7 vs. 29.4, p < .05). There was no statistically significant change in attitude toward breastfeeding (preimplementation score = 43.8, postimplementation score = 43.4, p = .075). For the mothers, the preintervention group reported exclusive breastfeeding 55% of the time versus 63% (p = .046) in the postintervention group. There was an increase in nurse observation of breastfeeding after the intervention: 84% versus 92% (p = .046), and breastfeeding patients reported an increase in nighttime breastfeeding after the intervention (55% vs. 71%, p = 0.001), with a resulting decrease in formula supplementation in the nursery at night (28% vs. 21%, p = .006). This study shows how an educational program and adherence to protocols can increase exclusive breastfeeding as well as improve healthcare provider knowledge, comfort level, and attitudes about breastfeeding. Our interventions also resulted in an increase in nurse observation of breastfeeding, nighttime breastfeeding, and a decrease in the use of formula supplementation at night. This study suggests that positive changes can be made with an educational program and protocols.


Contribution of dendritic cells to the autoimmune pathology of systemic lupus erythematosus.
Systemic lupus erythematosus (SLE) is a heterogeneous disease in which excessive inflammation, autoantibodies and complement activation lead to multisystem tissue damage. The contribution of the individual genetic composition has been extensively studied, and several susceptibility genes related to immune pathways that participate in SLE pathogenesis have been identified. It has been proposed that SLE takes place when susceptibility factors interact with environmental stimuli leading to a deregulated immune response. Experimental evidence suggests that such events are related to the failure of T-cell and B-cell suppression mediated by defects in cell signalling, immune tolerance and apoptotic mechanism promoting autoimmunity. In addition, it has been reported that dendritic cells (DCs) from SLE patients, which are crucial in the modulation of peripheral tolerance to self-antigens, show an increased ratio of activating/inhibitory receptors on their surfaces. This phenotype and an augmented expression of co-stimulatory molecules is thought to be critical for disease pathogenesis. Accordingly, tolerogenic DCs can be a potential strategy for developing antigen-specific therapies to reduce detrimental inflammation without causing systemic immunosuppression. In this review article we discuss the most relevant data relative to the contribution of DCs to the triggering of SLE.


[Diagnosis and treatment of orbital cellulitis].
Orbital cellulitis is a rare disease usually caused by sinusitis. Generally, the prognosis of both preseptal cellulitis and orbital cellulitis is favourable. Radiological distinction should be made between an orbital abscess and subperiosteal empyema. Theoretically, empyema is more accessible to antibiotics and, as it arises in a pre-existing anatomical space, it needs less aggressive treatment than an abscess. In contrast, the wall of an abscess created by the bacteria is scarcely permeable to antibiotics. Indications for surgical drainage should be based on clinical findings and not on Chandler's classification. Loss of vision, an unresponsive pupil or a densely packed orbit are indications for immediate surgical drainage. Drainage of an orbital abscess may speed up recovery.


Efficacy and safety of amoxycillin/clavulanate (Augmentin) twice daily versus three times daily in the treatment of acute otitis media in children. The Augmentin 454 Study Group.
This multicenter, randomized, single-blind study compared the efficacy and safety of a new, twice-daily formulation of amoxycillin/clavulanate (Augmenting) with the standard three-times-daily formulation. Children with a clinical diagnosis of acute otitis media, aged between 2 months and 12 years, received either amoxycillin/clavulanate 45/6.4 mg/kg/day twice-daily (b.d.) (range 38.3/5.5-76.2/10.9 mg/kg/day) or amoxycillin/clavulanate 40/10 mg/kg/day three-times-daily (t.d.s.) (range 25/6.25-56/14 mg/kg/day) for 7 or 10 days. Patients were evaluated during therapy (Days 3-5), at the end of therapy (Days 7-12) and at follow-up (Days 38-42). At the end of therapy, for the intent-to-treat and per-protocol populations, respectively, clinical success (cure) was achieved by approximately 94% of patients in both treatment groups. A successful bacteriological response at the end of therapy (Visit 3) was documented in 7/9 patients (77.8%) in the twice-daily group and in 11/13 patients (84.6%) in the three-times-daily group. At follow-up (Visit 4), 93.3% of patients in the twice-daily group and 87.9% in the three-times-daily group continued to have a clinically successful response. Both treatment regimens were well tolerated, with most adverse events being of a mild-moderate and transient nature. The most common treatment-related adverse event was diarrhea, occurring in 7.2% of patients in the twice-daily group and in 10.7% of the three-times-daily group. In total, 173 patients (82.8%) in the twice-daily group and 151 patients (73.3%) in the three-times-daily group were compliant with medication. In conclusion, this study confirms that b.d. amoxycillin/clavulanate is an effective treatment for pediatric acute otitis media and demonstrates that the b.d. and t.d.s. formulations of amoxycillin/clavulanate produce equivalent efficacy. Furthermore, there was a trend towards a higher level of compliance and a lower incidence of drug-related adverse events in the twice-daily compared with the three-times-daily treatment group.


Hsp60 in inflamed muscle tissue is the target of regulatory autoreactive T cells in patients with juvenile dermatomyositis.
Juvenile dermatomyositis (DM) is an autoimmune disease of unknown origin characterized by muscle weakness and skin manifestations. No definite autoantigen has yet been identified. Heat-shock proteins (HSPs) can be up-regulated at sites of inflammation, and immune reactivity to Hsp60 is suggested to play a regulatory role in various chronic inflammatory diseases. The purpose of this study was to determine whether Hsp60 could serve as an autoantigen in juvenile DM. Muscle tissue from 4 patients with juvenile DM and 1 healthy control subject without evidence of muscle disease was stained for Hsp60. Peripheral blood mononuclear cells (PBMCs) from 22 patients and 10 healthy control subjects were tested for T cell proliferation induced by human and microbial Hsp60. Cytokine production in response to Hsp60 was examined in 15 patients and 6 healthy controls. T cell reactivity to Hsp60 was determined in muscle biopsy samples from 2 patients. We found significantly increased T cell proliferation to human Hsp60 in PBMCs from juvenile DM patients, which was higher during disease remission. Following in vitro activation with Hsp60, significant amounts of tumor necrosis factor alpha, interleukin-1beta (IL-1beta), and IL-10 were produced. In contrast to muscle biopsy samples from healthy controls, samples from juvenile DM patients showed up-regulation of Hsp60, induction of T cell proliferation, and production of cytokines. Production of proinflammatory cytokines by muscle-derived cells in response to Hsp60 was associated with a poor clinical prognosis, whereas human Hsp60-specific induction of IL-10 was followed by clinical remission. These findings suggest that human (self) Hsp60 is a disease-relevant autoantigen in juvenile DM. The difference in T cell response with regard to disease activity indicates an immune regulatory effect of Hsp60-specific T cells, opening up perspectives for antigen-specific immunotherapy.


Isolation and hepatocyte differentiation of mesenchymal stem cells from porcine bone marrow--"surgical waste" as a novel MSC source.
Mesenchymal stem cells (MSC) isolated from bone marrow and differentiated into hepatocyte-like cells have increasingly gained attention for clinical cell therapy of liver diseases because of their high regenerative capacity. They are available from bone marrow aspirates of the os coxae after puncture of the crista iliaca or from bone marrow "surgical waste" gained from amputations or knee and hip operations. Thus, the aim of the study was to demonstrate whether these pBM-MSC (porcine bone marrow-derived mesenchymal stem cells) displayed mesenchymal features and hepatocyte differentiation potential. MSC were isolated either from crista iliaca punctures or after sampling and collagenase digestion of bone marrow from the os femoris. Mesenchymal features were assessed by flow cytometry for specific surface antigens and their ability to differentiate into at least 3 lineages. Functional properties, such as urea or glycogen synthesis and cytochrome P450 activity, as well as the cell morphology were examined during hepatocyte differentiation. pBM-MSC from both sources lacked the hematopoietic markers CD14 and CD45 but expressed the typical mesenchymal markers CD44, CD29, CD90, and CD105. Both cell types could differentiate into adipocyte, osteocyte, and hepatocyte lineages. After hepatocyte differentiation, CD105 expression decreased significantly and cells changed morphology from fibroblastoid into polygonal, displaying significantly increased glycogen storage, urea synthesis, and cytochrome activity. pBM-MSC from various sources were identical in respect to their mesenchymal features and their hepatocyte differentiation potential. Hence, long bones might be a particularly useful resource to isolate bone marrow mesenchymal stem cells for transplantation.


Patient-centered care: the jury is still out.
The patient-centered care model needs to retain a central focus on the patient. Process and system interfaces are key areas where alignment on behalf of the patient is required. Often, the current system is out of control. Departmental infrastructure and the need for resource reallocation must be assessed. No blueprint exists for implementing patient-centered care, although many incremental patient-focused initiatives are already underway. Impact on patients must be the balancing factor.


Anatomic distribution and pathologic characterization of small-volume prostate cancer (<0.5 ml) in whole-mount prostatectomy specimens.
Some investigators consider small-volume prostate cancer (0.5 ml or less) without Gleason pattern 4/5 elements as clinically insignificant. The objective of this study was to characterize the anatomic distribution and pathologic features of small tumors (aggregate volume of 0.5 ml or less) in whole-mount prostatectomy specimens. Between 1999 and 2003, 371 consecutive patients underwent radical prostatectomy at the Indiana University Hospitals for localized prostate cancer. Patients who received hormonal or radiation therapy prior to the surgery were excluded from the study. A total of 62 specimens with total tumor volume of 0.5 ml or less were identified and included in this study. All specimens were embedded and whole-mounted. Tumor volume was measured using the grid method. The mean age at the time of surgery was 59 years (median, 61 years; range, 37-72 years). The mean preoperative prostate-specific antigen (PSA) was 6.5 ng/ml (range: 0.3-18 ng/ml). The mean prostate weight was 53 g (range: 16-132 g). The mean tumor volume was 0.29 ml (median, 0.35 ml; range, 0.02-0.48 ml). Tumor multifocality and bilaterality were present in 69 and 37% of cases, respectively. Three (5%) had positive surgical margins. The largest tumor was located in the peripheral zone, transitional zone, and central zone in 79, 16, and 5% of cases, respectively. The largest tumor was located in the anterior prostate in 10 cases (16%) and in the posterior prostate in 52 cases (84%). The distribution of Gleason scores was 5 (12 cases, 19 %), 6 (40 cases, 65 %), and 7 (10 cases, 16 %). One case had a primary Gleason pattern 4. None had extraprostatic extension, seminal vesicle invasion, or lymph node metastasis. Small-volume prostate cancers are often multifocal and bilateral, with predilection for the peripheral zone. Of these small-volume cases, 16% had Gleason pattern 4 and might, therefore, be clinically significant.


Involvement of protein kinase C-alpha and -epsilon in extracellular Ca(2+) signalling mediated by the calcium sensing receptor.
The sensing of extracellular Ca(2+) concentration ([Ca(2+)](o)) and modulation of cellular processes associated with acute or sustained changes in [Ca(2+)](o) are cell-type specific and mediated by the calcium sensing receptor (CaR). [Ca(2+)](o) signalling requires protein kinase C (PKC), but the identity and role of PKC isoforms in CaR-mediated responses remain unclear. Here we show that high [Ca(2+)](o) activated PKC-alpha and PKC- in parathyroid cells and in human embryonic kidney (HEK293) cells overexpressing the CaR (HEK-CaR) and that this response correlated with the CaR-dependent activation of mitogen-activated protein kinases ERK1/2. Activation of ERK1/2 by acute high [Ca(2+)](o) required influx of Ca(2+)through Ni(2+)-sensitive Ca(2+)channels and phosphatidylinositol-dependent phospholipase C-beta activity. Inhibition of PKC by co-expression of dominant-negative (DN) mutants of PKC-alpha or - with the CaR attenuated sustained ERK1/2 activation. Overexpression of a PKC phosphorylation site (T888A) mutant CaR in HEK293 cells showed that this site was important for ERK1/2 activation at high [Ca(2+)](o). Activation of ERK1/2 by high [Ca(2+)](o) was not necessary for the [Ca(2+)](o)-regulated secretion of parathyroid hormone (PTH) in dispersed bovine parathyroid cells. These data suggest that the CaR-mediated [Ca(2+)](o) signal leading to regulated PTH secretion that requires diacylglycerol-responsive PKC isoforms is not mediated via the ERK pathway.


Metabolic engineering and transhydrogenase effects on NADPH availability in Escherichia coli.
The synthesis of several industrially useful compounds are cofactor-dependent, requiring reducing equivalents like NADPH in enzymatic reactions leading up to the synthesis of high-value compounds like polymers, chiral alcohols, and antibiotics. However, NADPH is costly and has limited intracellular availability. This study focuses on the study of the effect of the two transhydrogenase enzymes of Escherichia coli, PntAB and UdhA (SthA) on reducing equivalents-dependent biosynthesis. The production of (S)-2-chloropropionate from 2-chloroacrylate is used as a model system for monitoring NADPH availability because 2-haloacrylate reductase, the enzyme catalyzing the one-step conversion to (S)-2-chloropropionate in the synthesis pathway, requires NADPH as a cofactor. Results suggest that the presence of UdhA increases product yield and NADPH availability while the presence of PntAB has the opposite effect. A maximum product yield of 1.4 mol product/mol glucose was achieved aerobically in a pnt-deletion strain with udhA overexpression, a 150% improvement over the wild-type control strain.


Quality of life and treatment value in the management of hematologic malignancies.
This report is an overview of quality-of-life (QOL) concerns among people living with hematologic malignancies, and attempts to place these concerns in the context of overall therapeutic benefit and treatment value. There are two general categories of potential benefit offered by any treatment: time and QOL. If time can be added by a treatment, either by cure or prolonging survival, the value of that time makes toxicity more tolerable to patients, providers, payers, and society. If QOL can be improved, this will also make toxicity more acceptable. However, in the absence of time extension, this trade-off must demonstrate that the QOL benefit clearly outweighs the toxicity. In other words, the acceptability of toxicity increases as the likelihood of benefit improves. The range of possible symptoms and functional problems associated with hematologic malignancies is diverse, characterized by site and degree of involvement. Uncertainty of prognosis and the need to tolerate ambiguity frame many of the emotional challenges that face patients who must make difficult treatment and follow-up decisions. As more QOL data become available from clinical trials in hematologic malignancies, treatment decisions, while still difficult, can be made with more information about risk, type of improvements, toxicities, and probability of overall clinical benefit.


Percutaneous aspiration and alcohol sclerotherapy for symptomatic hepatic cysts. An alternative to surgical intervention.
Eight patients with 15 symptomatic nonneoplastic congenital hepatic cysts underwent ultrasound-guided percutaneous aspiration and temporary injection of 99% ethanol into the cyst. All cysts were treated at least twice at the same sitting. The volume of alcohol injected varied from 20 to 100 ml, depending on the size of the cyst. A cure was usually achieved with one alcohol sclerotherapy treatment. Only minor side effects such as transient pain and temperature elevation occurred. No recurrences were found during a follow-up period of 12 to 32 months. The results indicate that aspiration and alcohol sclerotherapy is a feasible alternative to surgical intervention in patients with symptomatic nonneoplastic congenital hepatic cysts. We recommend it as the treatment of choice in cases with high surgical risk or polycystic liver disease.


Myocardial Expression of Macrophage Migration Inhibitory Factor in Patients with Heart Failure.
Macrophage migration inhibitory factor (MIF) is a pleiotropic inflammatory protein and contributes to several different inflammatory and ischemic/hypoxic diseases. MIF was shown to be cardioprotective in experimental myocardial ischemia/reperfusion injury and its expression is regulated by the transcription factor hypoxia-inducible factor (HIF)-1α. We here report on MIF expression in the failing human heart and assess myocardial MIF in different types of cardiomyopathy. Myocardial tissue samples from n = 30 patients were analyzed by quantitative Real-Time PCR. MIF and HIF-1α mRNA expression was analyzed in myocardial samples from patients with ischemic (ICM) and non-ischemic cardiomyopathy (NICM) and from patients after heart transplantation (HTX). MIF expression was elevated in myocardial samples from patients with ICM compared to NICM. Transplanted hearts showed lower MIF levels compared to hearts from patients with ICM. Expression of HIF-1α was analyzed and was shown to be significantly increased in ICM patients compared to patients with NICM. MIF and HIF-1α mRNA is expressed in the human heart. MIF and HIF-1α expression depends on the underlying type of cardiomyopathy. Patients with ICM show increased myocardial MIF and HIF-1α expression.


Brugada phenocopies: Current evidence, diagnostic algorithms and a perspective for the future.
Brugada syndrome (BrS) is a congenital channelopathy associated with the development of malignant ventricular arrhythmias and sudden cardiac death. The diagnosis of BrS is made based on Brugada ECG pattern and clinical history. Brugada phenocopies (BrP) are clinical entities that are characterized by ECG patterns identical to those of BrS but arise from different underlying conditions such as metabolic abnormalities, myocardial ischemia and mechanical compression. Distinction between the two is important because BrS requires investigations for risk stratification whereas BrP requires appropriate treatment for the underlying conditions. In the present review, we summarized the current data in the literature, systematic diagnostic approach, gaps in the literature and future perspective on BrP.


Kramers problem: numerical Wiener-Hopf-like model characteristics.
Since the Kramers problem cannot be, in general, solved in terms of elementary functions, various numerical techniques or approximate methods must be employed. We present a study of characteristics for a particle in a damped well, which can be considered as a discretized version of the Melnikov [Phys. Rev. E 48, 3271 (1993)] turnover theory. The main goal is to justify the direct computational scheme to the basic Wiener-Hopf model. In contrast to the Melnikov approach, which implements factorization through a Cauchy-theorem-based formulation, we employ the Wiener-Levy theorem to reduce the Kramers problem to a Wiener-Hopf sum equation written in terms of Toeplitz matrices. This latter can provide a stringent test for the reliability of analytic approximations for energy distribution functions occurring in the Kramers problems at arbitrary damping. For certain conditions, the simulated characteristics are compared well with those determined using the conventional Fourier-integral formulas, but sometimes may differ slightly depending on the value of a dissipation parameter. Another important feature is that, with our method, we can avoid some complications inherent to the Melnikov method. The calculational technique reported in the present paper may gain particular importance in situations where the energy losses of the particle to the bath are a complex-shaped function of the particle energy and analytic solutions of desired accuracy are not at hand. In order to appreciate more readily the significance and scope of the present numerical approach, we also discuss concrete aspects relating to the field of superionic conductors.


Peptidylarginine deiminase 2 citrullinates MZB1 and promotes the secretion of IgM and IgA.
MZB1 is an endoplasmic reticulum residential protein preferentially expressed in plasma cells, marginal zone and B1 B cells. Recent studies on murine B cells show that it interacts with the tail piece of IgM and IgA heavy chain and promotes the secretion of these two classes of immunoglobulin. However, its role in primary human B cells has yet to be determined and how its function is regulated is still unknown. The conversion of peptidylarginine to peptidylcitrulline, also known as citrullination, by peptidylarginine deiminases (PADs) can critically influence the function of proteins in immune cells, such as neutrophils and T cells; however, the role of PADs in B cells remains to be elucidated. An unbiased analysis of human lung citrullinome was conducted to identify citrullinated proteins that are enriched in several chronic lung diseases, including rheumatoid arthritis-associated interstitial lung disease (RA-ILD), chronic obstructive pulmonary disease, and idiopathic pulmonary fibrosis, compared to healthy controls. Mass spectrometry, site-specific mutagenesis, and western blotting were used to confirm the citrullination of candidate proteins. Their citrullination was suppressed by pharmacological inhibition or genetic ablation of PAD2 and the impact of their citrullination on the function and differentiation of human B cells was examined with enzyme-linked immunosorbent assay, flow cytometry, and co-immunoprecipitation. Citrullinated MZB1 was preferentially enriched in RA-ILD but not in other chronic lung diseases. MZB1 was a substrate of PAD2 and was citrullinated during the differentiation of human plasmablasts. Ablation or pharmacological inhibition of PAD2 in primary human B cells attenuated the secretion of IgM and IgA but not IgG or the differentiation of IgM or IgA-expressing plasmablasts, recapitulating the effect of ablating MZB1. Furthermore, the physical interaction between endogenous MZB1 and IgM/IgA was attenuated by pharmacological inhibition of PAD2. Our data confirm the function of MZB1 in primary human plasmablasts and suggest that PAD2 promotes IgM/IgA secretion by citrullinating MZB1, thereby contributing to the pathogenesis of rheumatoid arthritis and RA-ILD.


Ultrashort Cell-Penetrating Peptides for Enhanced Sonophoresis-Mediated Transdermal Transport.
The skin is a key site for drug administration because of its large surface area and noninvasive accessibility. However, the dermal architecture serves as an excellent barrier, protecting from external mechanical, chemical, microbial, and physical perturbations. Most drugs display poor permeability through this barrier, thus making dermal and subdermal delivery challenging. Cell-penetrating peptides (CPPs), a diverse group of relatively short cationic and amphipathic membrane-interacting peptides, are fast becoming an important class of drug carriers and could potentially be developed for the dermal delivery of active molecules. However, the mechanism of CPP transdermal delivery is not fully understood, and there is a genuine need for a minimal model to understand this important phenomenon. Here, we demonstrate the potent membrane interactions of a minimal four-amino-acid-long CPP as well as the significance of guanidinium patterning and cationic nature of this palindromic peptide on its bioactivity. Furthermore, we demonstrate the biocompatibility of this peptide as well as its rapid cellular uptake and endosomal distribution. Finally, by utilizing a porcine full-thickness skin model, we demonstrate the substantial independent dermal and sonophoresis-based transdermal penetration of this minimal model. These results provide a minimal model for CPPs which can be easily manipulated for further biophysical and biochemical evaluations as well as a potent functional CPP with excellent skin permeability, which can be utilized for a wide variety of cosmetic and medical applications.


Sensory collectives in natural systems.
Groups of animals inhabit vastly different sensory worlds, or umwelten, which shape fundamental aspects of their behaviour. Yet the sensory ecology of species is rarely incorporated into the emerging field of collective behaviour, which studies the movements, population-level behaviours, and emergent properties of animal groups. Here, we review the contributions of sensory ecology and collective behaviour to understanding how animals move and interact within the context of their social and physical environments. Our goal is to advance and bridge these two areas of inquiry and highlight the potential for their creative integration. To achieve this goal, we organise our review around the following themes: (1) identifying the promise of integrating collective behaviour and sensory ecology; (2) defining and exploring the concept of a 'sensory collective'; (3) considering the potential for sensory collectives to shape the evolution of sensory systems; (4) exploring examples from diverse taxa to illustrate neural circuits involved in sensing and collective behaviour; and (5) suggesting the need for creative conceptual and methodological advances to quantify 'sensescapes'. In the final section, (6) applications to biological conservation, we argue that these topics are timely, given the ongoing anthropogenic changes to sensory stimuli (e.g. via light, sound, and chemical pollution) which are anticipated to impact animal collectives and group-level behaviour and, in turn, ecosystem composition and function. Our synthesis seeks to provide a forward-looking perspective on how sensory ecologists and collective behaviourists can both learn from and inspire one another to advance our understanding of animal behaviour, ecology, adaptation, and evolution.


Dynamics of unfolded nucleosomal fiber.
The "rigidity" of chromatin fiber solenoidal structure in different states of condensation was evaluated with the help of gel-electrophoresis. A new property of the unfolded nucleosomal fiber-the capacity to condense with temperature-was demonstrated. These results together with our previously obtained data (W.A. Krajewski et al., Mol. Gen. Genet. 230, pp. 442-448, 1991; W.A. Krajewski et al., Ibid. 231, pp. 17-22, 1991) testify that changes in DNA linking number of transcriptionally active minichromosomes arise in vivo from alteration of nucleosomal solenoid parameters (i.e. from supernucleosomal level of chromatin organization), rather than from core histone modifications only or from increased flexibility of DNA within nucleosomes.


[Influence of the container and environmental factors in the stability of sodium hypochlorite].
This study was performed to analyze the chemical stability of sodium hypochlorite (with 2.6% or 26.0 mg/mL of free residual chlorine) in the various types of containers when exposed to different light levels. Over a period of eight months, several samples of that solution were put into containers made of glass (amber or clear) and plastic (opaque white, opaque green, or clear) with and without a top, and were exposed to three different lighting conditions (continuous light, total dark, and ambient light of the laboratory where the study was carried out). The stability of the solution was analyzed by measuring the concentration of free residual chlorine in the samples exposed as described above. The environmental factors tested were chosen as representative of the various conditions under which this effective, cheap, and widely available household disinfectant is often stored and used by the general public. The results indicated that chemical instability increased with the presence of light, the absence of a top, and the length of time the solution was stored. The most suitable containers were those made of amber glass, opaque green plastic, or opaque white plastic, although the last one offered the least protection of the three.


Estrogen receptor α gene polymorphisms and risk of Alzheimer's disease: evidence from a meta-analysis.
Human estrogen receptor α (ESR1), a member of the nuclear receptor superfamily of ligand-activated transcription factors, is one of the key mediators of hormonal response in estrogen-sensitive tissues. Accumulating evidence has demonstrated that two of the most widely studied single-nucleotide polymorphisms in ESR1 - PvuII (T/C, rs223493) and Xbal (A/G, rs9340799) - are possibly associated with Alzheimer's disease (AD). However, individual study results are still controversial. We searched PubMed, Embase, Web of Science, Science Direct, SpringerLink, and the Chinese National Knowledge Infrastructure databases for eligible studies assessing the association of ESR1 polymorphisms and AD risk (last search performed in November 2013). Thereafter, a meta-analysis of 13,192 subjects from 18 individual studies was conducted to evaluate the association between ESR1 polymorphisms and susceptibility to AD. The results indicated that a significant association was found between the ESR1 PvuII polymorphism and AD risk in Caucasian populations (CC + CT versus TT, odds ratio [OR] 1.14, 95% confidence interval [CI] 1.02-1.28, P=0.03; CT versus TT, OR 1.16, 95% CI 1.02-1.31, P=0.02), whereas no evidence of association was found in Asian populations. Nevertheless, we did not find any significant association between the ESR1 XbaI polymorphism and AD risk for any model in Caucasian and Asian populations (all P>0.05). Based on this meta-analysis, we conclude that the ESR1 PvuII polymorphism might be a risk factor in AD development in Caucasian populations, not in Asian populations. Further confirmation is needed from better-designed and larger studies.


Beta 2-agonist-elevated stress response in human bronchial epithelial cells in vivo and in vitro.
Recent studies report high baseline levels of stress (heat shock) proteins in bronchial epithelial cells from asthmatic individuals. The promoter of the gene encoding the 72-kDa heat shock protein has an element responsive to cAMP, which may be affected by beta-agonists. This study examined stress protein levels in subjects enrolled in a segmental lung allergen challenge study to determine whether beta-agonist medication could contribute to a stress response. Subjects were divided on the basis of no premedication (n = 17), metered dose inhalations of albuterol (n = 24), or placebo inhalation (n = 3) prior to bronchoscopy. Levels of the inducible stress protein Hsp72 and constitutive Hsp73 were quantitated in bronchial epithelial cells from brush biopsy of allergic nonasthmatic, allergic asthmatic, and normal individuals. Mean levels were increased significantly (p < 0.003 and p < 0.004, respectively) in those subjects who received albuterol premedication. No significant differences were found between clinical groups of individuals or for placebo inhalation vs nonpremedication. Albuterol in vitro increased the levels of Hsp72 and Hsp73 in epithelial cells from either nonpremedicated or placebo-treated donors; the Hsp72 levels correlated linearly with increased albuterol concentration (r = 0.81, p < 0.01). Therefore, beta-agonists elevate or prolong an elevated stress response in epithelial cells, possibly through cAMP-mediated effects.


The autoimmune encephalitis-related cytokine TSLP in the brain primes neuroinflammation by activating the JAK2-NLRP3 axis.
NLRP3 inflammasome hyperactivation contributes to neuroinflammation in autoimmune disorders, but the underlying regulatory mechanism remains to be elucidated. We demonstrate that compared with wild-type (WT) mice, mice lacking thymic stromal lymphopoietin (TSLP) receptor (TSLPR) (Tslpr-/- mice) exhibit a significantly decreased experimental autoimmune encephalomyelitis (EAE) score, reduced CD4+ T cell infiltration, and restored myelin basic protein (MBP) expression in the brain after EAE induction by myelin oligodendrocyte glycoprotein35-55 (MOG35-55). TSLPR signals through Janus kinase (JAK)2, but not JAK1 or JAK3, to induce NLRP3 expression, and Tslpr-/- mice with EAE show decreased JAK2 phosphorylation and NLRP3 expression in the brain. JAK2 inhibition by ruxolitinib mimicked loss of TSLPR function in vivo and further decreased TSLP expression in the EAE mouse brain. The NLRP3 inhibitor MCC950 decreased CD4+ T cell infiltration, restored MBP expression, and decreased IL-1β and TSLP levels, verifying the pro-inflammatory role of NLRP3. In vitro experiments using BV-2 murine microglia revealed that TSLP directly induced NLRP3 expression, phosphorylation of JAK2 but not JAK1orJAK3, and IL-1β release, which were markedly inhibited by ruxolitinib. Furthermore, EAE induction led to an increase in the Th17 cell number, a decrease in the regulatory T (Treg) cell number in the blood, and an increase in the expression of the cytokine IL-17A in the WT mouse brain, which was drastically reversed in Tslpr-/- mice. In addition, ruxolitinib suppressed the increase in IL-17A expression in the EAE mouse brain. These findings identify TSLP as a prospective target for treating JAK2-NLRP3 axis-associated autoimmune inflammatory disorders.


A Novel Magnetometer Array-based wearable system for ASL gesture recognition.
Hand gesture classification is of high importance in any sign language recognition (SLR) system, which is expected to assist individuals suffering from hearing and speech impairment. American sign language (ASL) comprises of static and dynamic gestures representing many alphabets, phrases, and words. ASL recognition system allows us to digitize communication and use it effectively within or outside the hearing-deprived community. Developing an ASL recognition system has been a challenge since some of the involved hand gestures closely resemble each other, and thereby it demands high discriminability features to classify these gestures. SLR through surface-based electromyography (sEMG) signals is computationally intensive to process and using inertial measurement units (IMUs) or flex sensors for SLR occupies too much space on the patient's hand. Video-based recognition systems place restrictions on the users by requiring them to make gestures or motions within the camera's field of view. A novel approach with a precision preserved static gesture classification system is proposed to fulfill the much-needed gap. The paper proposes an array of magnetometers-enabled static hand gesture classification system that offers an average accuracy of 98.60% for classifying alphabets and 94.07% for digits using the KNN classification model. The magnetometer array-based wearable system is devised to minimize the electronics coverage around the hand, and yet establish robust classification results that are useful for ASL recognition. The paper discusses the design of the proposed SLR system and also looks into optimizations that can be made to reduce the cost of the system.Clinical relevance - The proposed novel magnetometer array-based wearable system is cost-effective and works well across different hand sizes. It occupies a negligible amount of space on the user's hand and thus does not interfere with the user's everyday tasks. It is reliable, robust, and error-free for easy adoption towards building ASL recognition system.


Cumulative risk assessment of pesticide residues in food.
There is increasing need to address the potential risks of combined exposures to multiple residues from pesticides in the diet. The available evidence suggests that the main concern is from dose addition of those compounds that act by the same mode of action. The possibility of synergy needs to be addressed on a case-by-case basis, where there is a biologically plausible hypothesis that it may occur at the levels of residues occurring in the diet. Cumulative risk assessment is a resource-intense activity and hence a tiered approach to both toxicological evaluation and intake estimation is recommended, and the European Food Safety Authority (EFSA) has recently published such a proposal. Where assessments have already been undertaken by some other authority, full advantage should be taken of these, subject of course to considerations of quality and relevance. Inclusion of compounds in a cumulative assessment group (CAG) should be based on defined criteria, which allow for refinement in a tiered approach. These criteria should include chemical structure, mechanism of pesticidal action, target organ and toxic mode of action. A number of methods are available for cumulating toxicity. These are all inter-related, but some are mathematically more complex than others. The most useful methods, in increasing levels of complexity and refinement, are the hazard index, the reference point index, the Relative Potency Factor method and physiologically based toxicokinetic modelling, although this last method would only be considered should a highly refined assessment be necessary. Four possible exposure scenarios are of relevance for cumulative risk assessment, acute and chronic exposure in the context of maximum residue level (MRL)-setting, and in relation to exposures from the actual use patterns, respectively. Each can be addressed either deterministically or probabilistically. Strategies for dealing with residues below the limit of detection, limit of quantification or limit of reporting need to be agreed. A number of probabilistic models are available, but some of there are geographically constrained due to the underlying datasets used in their construction. Guidance on probabilistic modelling needs to be finalised. Cumulative risk assessments have been performed in a number of countries, on organophosphate insecticides alone (USA) or together with carbamates (UK, DK, NL), triazines, chloroacetanilides, carbamates alone (USA), and all pesticides (DE). All identifiable assumptions and uncertainties should be tabulated and evaluated, at least qualitatively. Those likely to have a major impact on the outcome of the assessment should be examined quantitatively. In cumulative risk assessment, it is necessary, as in other risk assessments, for risk managers to consider what level of risk would be considered "acceptable", for example what percentile of the population should be below the reference value. Criteria for prioritising CAGs for cumulative risk assessment include frequency of detection in monitoring programmes, high usage, high exposure relative to the reference value, large number of compounds (e.g. five or more) in a group.


The Use of Photofunctionalized Implants for Low or Extremely Low Primary Stability Cases.
The objective of this study was to examine the success rate and development of implant stability of photofunctionalized implants with low and extremely low primary stability. Primary implant stability was evaluated by measuring implant stability quotients (ISQs) at placement (initial ISQ). Of 301 consecutive implants placed over 2 years, 55 implants (in 38 patients) with initial ISQs less than 60 were included. All implants were photofunctionalized by chairside treatment with ultraviolet light for 15 minutes using a photo device immediately prior to placement, and the generation of superhydrophilicity and hemophilicity was confirmed. The rate of implant stability development was evaluated by calculating the osseointegration speed index (OSI), defined as the ISQ increase per month. Most of the 55 implants (90.9%) were in complex cases and placed in site-developmental sites with simultaneous or staged guided bone regeneration and/or sinus elevation or fresh extraction sites. The success rate was 98.2% after 2 to 3 years of follow-up. The ISQ for all implants was 50.4 ± 7.7 at placement and 74.3 ± 5.7 at stage-two surgery (P < .001) after an average of 7.1 months of healing. All ISQ values were greater than 60 at the stage-two surgery. ISQ increases were inversely correlated with initial ISQ (R = 0.789) and resulted in consistently high ISQs at the stage-two surgery regardless of the degree of the initial ISQ. Implants with absent primary stability for which ISQ measurement was not possible at placement also attained ISQs of 75 or greater at the stage-two surgery. The OSIs of low initial stability (55 or lower) implants ranged from 3.9 to 4.7, which was considerably higher than the OSIs of as-received implants reported in the literature (0.36 to 2.8). Photofunctionalized implants placed with low, extremely low, or absent primary stability showed a high success rate, which was corroborated with a rapid increase in implant stability during the healing. Photofunctionalization was more effective for implants with lower primary stability, indicating that the technology was particularly effective in mitigating the challenging conditions.


Evaluation of behavior and development training for pediatric residents.
The evaluation of a behavior and development training program that was integrated into a general pediatric training residency is reported. Thirty-two residents who participated in the program were evaluated over one year on measures designed to assess residents' opinions about the relevance of behavior and development concepts, residents' perceptions of their own competence in behavior and development knowledge and skills, supervisors' ratings of residents' general clinical performance, and residents' attitudes about 10 specific illnesses. Results suggest that residents perceive a high relevance for psychosocial concepts and an increasing competence with these concepts after training. Establishing rapport with patients emerged as an important variable in supervisors' ratings of resident performance and in residents' self-ratings of competence. Attitudes were initially negative toward the more behavioral and psychophysiological disease entities but showed some positive change over the year. These results and their implications are discussed.


Resonance Analysis and Gain Estimation Using CMA-Based Even Mode Combination Method for Flexible Wideband Antennas.
This work presents an efficient design and optimization method based on characteristic mode analysis (CMA) to predict the resonance and gain of wideband antennas made from flexible materials. Known as the even mode combination (EMC) method based on CMA, the forward gain is estimated based on the principle of summing the electric field magnitudes of the first even dominant modes of the antenna. To demonstrate its effectiveness, two compact, flexible planar monopole antennas designed on different materials and two different feeding methods are presented and analyzed. The first planar monopole is designed on Kapton polyimide substrate and fed using a coplanar waveguide to operate from 2 to 5.27 GHz (measured). On the other hand, the second antenna is designed on felt textile and fed using a microstrip line to operate from about 2.99 to 5.57 GHz (measured). Their frequencies are selected to ensure their relevance in operating across several important wireless frequency bands, such as 2.45 GHz, 3.6 GHz, 5.5 GHz, and 5.8 GHz. On the other hand, these antennas are also designed to enable competitive bandwidth and compactness relative to the recent literature. Comparison of the optimized gains and other performance parameters of both structures are in agreement with the optimized results from full wave simulations, which process is less resource-efficient and more iterative.


Characterization of inward-rectifier K+ channel inhibition by antiarrhythmic piperazine.
Strong inward-rectifier K(+) (Kir) channels play a significant role in shaping the cardiac action potential: they help produce its long plateau and accelerate its rate of repolarization. Consequently, genetic deletion of the gene encoding the strongly rectifying K(+) channel IRK1 (Kir2.1) prolongs the cardiac action potential in mice. In principle, broadening the action potential lengthens the refractory period, which may in turn be antiarrhythmogenic. Interestingly, previous studies showed that piperazine, an inexpensive and safe anthelmintic, both inhibits IRK1 channels and is antiarrhythmic in some animal preparations. This potential pharmacological benefit motivated us to further characterize the energetic, kinetic, and molecular properties of IRK1 inhibition by piperazine. We show how its blocking characteristics, in particular, its shallow voltage dependence, allow piperazine to be effective even in the presence of high-affinity polyamine blockers. We also examine the channel selectivity of piperazine and its molecular determinants.


Patched regulation of axon guidance is by specifying neural identity in the Drosophila nerve cord.
Within an axon bundle, one or two are pioneering axons and the rest are follower axons. Pioneering axons are projected first and the follower axons are projected later but follow a pioneering axon(s) pathway. It is not clear whether the pioneering axons have a guidance role for follower axons. In this paper, we have investigated the role of Patched (Ptc) in regulating the guidance of medial tract, one of the longitudinal tracts in the nerve cord. In patched mutants the medial longitudinal tract fails to fasciculate on its own side along the nerve cord, instead it abnormally crosses the midline and fasciculates with the contralateral tract. Interestingly, the medial tracts cross the midline ignoring the axon-repellant Slit on the midline and Roundabout on growth cones. The medial tract is pioneered by neurons pCC and vMP2. Our results show that guidance defects of this tract are due to loss and mis-specification of vMP2, which results in the projection from pCC to either stall or project outward near the location of vMP2. Thus, both pioneering neurons are necessary for the proper guidance of pioneering and follower axons. We also show that the loss of Ptc activity in the neuroectoderm prior to the formation of S1 and S2 neuroblasts causes the majority of axon guidance defects. These results provide insight into how mis-specification and loss of neurons can non-autonomously contribute to defects in axon pathfinding.


Minimizing superficial thermal injury using bilateral cryogen spray cooling during laser reshaping of composite cartilage grafts.
Composite cartilage grafts were excised from New Zealand rabbit ears. Flat composite grafts (of cartilage and overlying skin graft on both surfaces) were obtained from each ear and cut into a rectangle measuring 50 mm by 25 mm (x by y) with an average thickness of approximately 1.3 mm (z), skin included. Specimens were manually deformed with a jig and maintained in this new position during laser illumination. The composite cartilage grafts were illuminated on the concave surface with an Nd:YAG laser (1,064 nm, 3 mm spot) at 10 W, 20 W, 30 W, 40 W, 50 W. Cryogen spray cooling (CSC) was applied to both exterior (convex) and interior (concave) surfaces of the tissue to reduce thermal injury to the grafts. CSC was delivered: (1) in controlled applications (cryogen released when surface reached 40 degrees C, and (2) receiving only laser at above wattage, no CSC [representing the control group]. The specimens were maintained in a deformation for 15 minutes after illumination and serially examined for 14 days. The control group with no CSC caused injury to all specimens, ranging from minor to full thickness epidermal thermal injury. Although most levels of laser and CSC yielded a high degree of reshaping over an acute time period, after 14 days specimens exposed to 30 W, 40 W, 50 W retained shape better than those treated at 10 W and 20 W. The specimens exposed to 50 W with controlled CSC retained its new shape to the highest degree over all others, and thermal injury was minimal. In conclusion, combinations of laser and CSC parameters were effective and practical for the reshaping of composite cartilage grafts.


Three Novel EGFR Mutations (750_758del, I759S, T751_I759delinsS) in One Patient with Metastatic Non-Small Cell Lung Cancer Responding to Osimertinib: A Case Report.
Generations of epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) can significantly improve the outcome of EGFR-positive NSCLC patients. However, acquired TKIs-resistant mutations are inevitable. Except the common EGFR alterations, more and more rare mutations are revealed by next-generation sequencing (NGS), the clinical significance of which are still unclear. Here, we report an advanced lung adenocarcinoma patient who harbored two novel EGFR exon 19 deletions (750_758del and I759S) at the beginning and exhibited a short response to icotinib for 7.0 months. Then, secondary resistance EGFR T751_I759delinsS occurred. Chemotherapy combined with bevacizumab and erlotinib was administered in turn but failed. Standard-dose osimertinib (80 mg daily) obtained durable clinical remission for 16 months, and high-dose osimertinib (160 mg daily) further prolonged the survival of 9 months after leptomeningeal metastases (LM) occurring. This study presented the first case of intractable terminal NSCLC in a patient with EGFR 750_758del, I759S and T751_I759delinsS mutations, who responded positively to osimertinib and achieved a prolonged OS of 52 months, providing a potential therapeutic option for the patients harboring these particular EGFR mutations.


Mechanical strain leads to condylar growth in adult rats.
Mechanical strain produced by forward mandibular positioning was found to enhance mandibular condylar growth in experimental animals and in patients. This study was designed to identify the changes in number and rate of the proliferating mesenchymal cells in mandibular condyles of adult rats and to correlate these changes to the expression of SOX9 and type II collagen under mechanical strain. Seventy-eight 120-day-old female Sprague-Dawley rats were randomly allotted to six groups, nine animals in each experimental group according to different time points. Cell kinetic studies for expression of PCNA were used to identify number and rate of proliferating mesenchymal cells. Immunostaining of SOX9 and in situ hybridization of Col2a1 gene were carried out. Results showed a significant increase in number of replicating mesenchymal cells and proliferation rate. The expression of SOX9 was enhanced and Col2a1 gene transcript was then activated. The proliferative layer became thicker on experimental day 21. The thickness of chondroblast layer and chondrocyte layer showed significant increase from experimental day 14 to day 30. In conclusion, mechanical strain produced by mandibular advancement in adult rats promotes the proliferation of mesenchymal cells. Under control of transcription factor SOX9, these mesenchymal cells are then committed to enter the chondrogenic route leading to condylar growth.


The adjuvant activity of alphavirus replicons is enhanced by incorporating the microbial molecule flagellin into the replicon.
Ligands of pattern recognition receptors (PRRs) including Toll-like receptors (TLRs) stimulate innate and adaptive immune responses and are considered as potent adjuvants. Combinations of ligands might act in synergy to induce stronger and broader immune responses compared to stand-alone ligands. Alphaviruses stimulate endosomal TLRs 3, 7 and 8 as well as the cytoplasmic PRR MDA-5, resulting in induction of a strong type I interferon (IFN) response. Bacterial flagellin stimulates TLR5 and when delivered intracellularly the cytosolic PRR NLRC4, leading to secretion of proinflammatory cytokines. Both alphaviruses and flagellin have independently been shown to act as adjuvants for antigen-specific antibody responses. Here, we hypothesized that alphavirus and flagellin would act in synergy when combined. We therefore cloned the Salmonella Typhimurium flagellin (FliC) gene into an alphavirus replicon and assessed its adjuvant activity on the antibody response against co-administered antigen. In mice immunized with recombinant alphavirus, antibody responses were greatly enhanced compared to soluble FliC or control alphavirus. Both IgG1 and IgG2a/c responses were increased, indicating an enhancement of both Th1 and Th2 type responses. The adjuvant activity of FliC-expressing alphavirus was diminished but not abolished in the absence of TLR5 or type I IFN signaling, suggesting the contribution of several signaling pathways and some synergistic and redundant activity of its components. Thus, we have created a recombinant adjuvant that stimulates multiple signaling pathways of innate immunity resulting in a strong and broad antibody response.


Targeting MDR1 gene: synthesis and cellular study of modified daunomycin-triplex-forming oligonucleotide conjugates able to inhibit gene expression in resistant cell lines.
Reversal of the multidrug-resistant (MDR) phenotype is very important for chemotherapy success. In fact, the expression of the MDR1 gene-encoded P-glycoprotein (P-gp) actively expels antitumor agents such as daunomycin (DNM) out of the cells, resulting in drug resistance. We show that upon conjugation to triplex-forming oligonucleotides, it is possible to address DNM in resistant cells (MCF7-R and NIH-MDR-G185). The oligonucleotide moiety of the conjugate changes the cellular penetration properties of the antitumor agent that is no more the target of P-gp in resistant cells. We observe an accumulation of conjugated DNM in cells up to 72 h. For more efficient delivery in the cells' nuclei, transfectant agents must be used. In addition, the conjugate recognizes a sequence located in exon 3 of MDR1, and it inhibits its gene expression as measured both by Western blot and by reverse transcription-polymerase chain reaction.


A versatile in vivo system for directed dissection of gene expression patterns.
Tissue-specific gene expression using the upstream activating sequence (UAS)–GAL4 binary system has facilitated genetic dissection of many biological processes in Drosophila melanogaster. Refining GAL4 expression patterns or independently manipulating multiple cell populations using additional binary systems are common experimental goals. To simplify these processes, we developed a convertible genetic platform, the integrase swappable in vivo targeting element (InSITE) system. This approach allows GAL4 to be replaced with any other sequence, placing different genetic effectors under the control of the same regulatory elements. Using InSITE, GAL4 can be replaced with LexA or QF, allowing an expression pattern to be repurposed. GAL4 can also be replaced with GAL80 or split-GAL4 hemi-drivers, allowing intersectional approaches to refine expression patterns. The exchanges occur through efficient in vivo manipulations, making it possible to generate many swaps in parallel. This system is modular, allowing future genetic tools to be easily incorporated into the existing framework.


Gene context analysis reveals functional divergence between hypothetically equivalent enzymes of the purine-ureide pathway.
A major problem of genome annotation is the assignment of a function to a large number of genes of known sequences through comparison with a relatively small number of experimentally characterized genes. Because functional divergence is a widespread phenomenon in gene evolution, the transfer of a function to homologous genes is not a trivial exercise. Here, we show that a family of homologous genes which are found in purine catabolism clusters and have hypothetically equivalent functions can be divided into two distinct groups based on the genomic distribution of functionally related genes. One group (UGLYAH) encodes proteins that are able to release ammonia from (S)-ureidoglycine, the enzymatic product of allantoate amidohydrolase (AAH), but are unable to degrade allantoate. The presence of a gene encoding UGLYAH implies the presence of AAH in the same genome. The other group (UGLYAH2) encodes proteins that are able to release ammonia from (S)-ureidoglycine as well as urea from allantoate. The presence of a gene encoding UGLYAH2 implies the absence of AAH in the same genome. Because (S)-ureidoglycine is an unstable compound that is only formed by the AAH reaction, the in vivo function of this group of enzymes must be the release of urea from allantoate (allantoicase activity), while ammonia release from (S)-ureidoglycine is an accessory activity that evolved as a specialized function in a group of genes in which the coexistence with AAH was established. Insights on the active site modifications leading to a change in the enzyme activity were provided by comparison of three-dimensional structures of proteins belonging to the two different groups and by site-directed mutagenesis. Our results indicate that when the neighborhood of uncharacterized genes suggests a role in the same process or pathway of a characterized homologue, a detailed analysis of the gene context is required for the transfer of functional annotations.


Stroke patterns and regulation of swim speed and energy cost in free-ranging Brünnich's guillemots.
Loggers were attached to free-ranging Brünnich's guillemots Uria lomvia during dives, to measure swim speeds, body angles, stroke rates, stroke and glide durations, and acceleration patterns within strokes, and the data were used to model the mechanical costs of propelling the body fuselage (head and trunk excluding wings). During vertical dives to 102-135 m, guillemots regulated their speed during descent and much of ascent to about 1.6+/-0.2 m s(-1). Stroke rate declined very gradually with depth, with little or no gliding between strokes. Entire strokes from 2 m to 20 m depth had similar forward thrust on upstroke vs downstroke, whereas at deeper depths and during horizontal swimming there was much greater thrust on the downstroke. Despite this distinct transition, these differences had small effect (<6%) on our estimates of mechanical cost to propel the body fuselage, which did not include drag of the wings. Work stroke(-1) was quite high as speed increased dramatically in the first 5 m of descent against high buoyancy. Thereafter, speed and associated drag increased gradually as buoyancy slowly declined, so that mechanical work stroke(-1) during the rest of descent stayed relatively constant. Similar work stroke(-1) was maintained during non-pursuit swimming at the bottom, and during powered ascent to the depth of neutral buoyancy (about 71 m). Even with adjustments in respiratory air volume of +/-60%, modeled work against buoyancy was important mainly in the top 15 m of descent, after which almost all work was against drag. Drag was in fact underestimated, as our values did not include enhancement of drag by altered flow around active swimmers. With increasing buoyancy during ascent above 71 m, stroke rate, glide periods, stroke acceleration patterns, body angle and work stroke(-1) were far more variable than during descent; however, mean speed remained fairly constant until buoyancy increased rapidly near the surface. For dives to depths >20 m, drag is by far the main component of mechanical work for these diving birds, and speed may be regulated to keep work against drag within a relatively narrow range.


Significant genetic differentiation of Gobiopterus lacustris, a newly recorded transparent goby in China.
In our recent survey, the transparent small Lacustrine goby, Gobiopterus lacustris had reported as the endemic species of Luzon, Philippines, was identified as an abundant species in mangroves of Leizhou Peninsula, China. Here, high diversity and significant differentiation of five sites of samples representing the west and east populations were revealed by mitochondrial DNA sequences. Five haplotypes of 56 cytochrome oxidase subunit I (Cox1) with the lengths of 623 base pairs (bp) have the high pairwise identity (>98.8%). Moreover, a total of 31 haplotypes for 129 partial D-loop regions were clustered into two clades corresponding to the east and west sampling sites. The strong population structure was confirmed (ΦST = 0.43017, p < .0001) with high haplotype diversity (h = 0.880 ± 0.017) and low nucleotide diversity (p=.00484). Moreover, both the mismatch distribution analysis and neutral test of D-loop revealed that the west group might experience a recent demographic expansion. Lastly, the isolation-with-migration analysis supported the expansion and indicated that the east-west split happened at approximately 7.1 kyr ago. Given the distribution and diversity, G. lacustris could be a good model for the study of the sea-level fluctuations and coast evolution of the South China Sea.


Aldose Reductase Mediates NLRP3 Inflammasome-Initiated Innate Immune Response in Hyperglycemia-Induced Thp1 Monocytes and Male Mice.
Despite recent studies that show oxidative stress-generated reactive oxygen species (ROS) regulate NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome-mediated innate immune response in various diabetic complications, the mechanism by which ROS activate innate immune response is not well understood. We have shown previously that aldose reductase (AR), besides reducing glucose, reduces lipid aldehydes and their glutathione conjugates and participates in various oxidative stress-induced inflammatory pathways. To understand the role of AR in ROS-induced innate immune response, we have investigated the mechanism(s) by which AR activates hyperglycemia-induced NLRP3 inflammsome-initiated innate immune response in Thp1 monocytes and in streptozotocin (STZ)-induced diabetic mice. In Thp1 monocytes, inhibition or ablation of AR prevented high-glucose-induced activation of NLRP3 inflammasome and caspase-1 and release of the innate immune cytokines interleukin (IL)-1β and IL-18. AR inhibition in Thp1 cells also prevented the high-glucose-induced generation of ROS, influx of Ca2+, efflux of K+, and activation of Lyn, Syk, and PI3K. Furthermore, the AR inhibitor fidarestat prevented the expression of NLRP inflammasome components in STZ-induced diabetic mouse heart and aorta, and also prevented the release of various cytokines in the serum. Collectively, our data suggest that AR regulates hyperglycemia-induced NLRP3 inflammasome-mediated innate immune response by altering the ROS/Lyn/Syk/PI3K/Ca2+/K+ signals.


[Mechanism of the activating action of noradrenaline on rat cerebral cortex Na, K-ATPase].
Effect of noradrenaline on kinetics of Na, K-ATPase activation in rat brain cortex was studied using ATP as a substrate and Na+ or K+ ions as cofactors. Noradrenaline (5 x 10(-5) M) increased the activating effect of ATP and Mg2+ as well as that of Na+ and K+. In presence of ATP and Mg2+ the hormone activated ATPase by the mixed type, and in response to Na+, K+ addition--by the non-competitive type. Noradrenaline appears to cause the conformational alterations in Na, K-ATPase molecule, which change the kinetics of the enzyme activation by ATP and the ions.


Seroprevalence and associated risk factors of chikungunya, dengue, and Zika in eight districts in Tanzania.
This study was conducted to determine the seroprevalence and risk factors of chikungunya (CHIKV), dengue (DENV), and Zika (ZIKV) viruses in Tanzania. The study covered the districts of Buhigwe, Kalambo, Kilindi, Kinondoni, Kondoa, Kyela, Mvomero, and Ukerewe in Tanzania. Blood samples were collected from individuals recruited from households and healthcare facilities. An ELISA was used to screen for immunoglobulin G antibodies against CHIKV, DENV, and ZIKV. A total of 1818 participants (median age 34 years) were recruited. The overall CHIKV, DENV, and ZIKV seroprevalence rates were 28.0%, 16.1%, and 6.8%, respectively. CHIKV prevalence was highest in Buhigwe (46.8%), DENV in Kinondoni (43.8%), and ZIKV in Ukerewe (10.6%) and Mvomero (10.6%). Increasing age and frequent mosquito bites were significantly associated with CHIKV and DENV seropositivity (P < 0.05). Having piped water or the presence of stagnant water around the home (P < 0.01) were associated with higher odds of DENV seropositivity. Fever was significantly associated with increased odds of CHIKV seropositivity (P < 0.001). Visiting mines had higher odds of ZIKV seropositivity (P < 0.05). These findings indicate that DENV, CHIKV, and ZIKV are circulating in diverse ecological zones of Tanzania. There is a need to strengthen the control of mosquito-borne viral diseases in Tanzania.


Impact of perioperative acute ischemic stroke on the outcomes of noncardiac and nonvascular surgery: a single centre prospective study.
Although ischemic stroke is a well-known complication of cardiovascular surgery it has not been extensively studied in patients undergoing noncardiac surgery. The aim of this study was to assess the predictors and outcomes of perioperative acute ischemic stroke (PAIS) in patients undergoing noncardiothoracic, nonvascular surgery (NCS). We prospectively evaluated patients undergoing NCS and enrolled patients older than 18 years who underwent an elective, non-daytime, open surgical procedure. Electrocardiography and cardiac biomarkers were obtained 1 day before surgery, and on postoperative days 1, 3 and 7. Of the 1340 patients undergoing NCS, 31 (2.3%) experienced PAIS. Only age (odds ratio [OR] 2.5, 95% confidence interval [CI] 1.01-3.2, p < 0.001) and preoperative history of stroke (OR 3.6, 95% CI 1.2-4.8, p < 0.001) were independent predictors of PAIS according to multivariate analysis. Patients with PAIS had more cardiovascular (51.6% v. 10.6%, p < 0.001) and noncardiovascular complications (67.7% v. 28.3%, p < 0.001). In-hospital mortality was 19.3% for the PAIS group and 1% for those without PAIS (p < 0.001). Age and preoperative history of stroke were strong risk factors for PAIS in patients undergoing NCS. Patients with PAIS carry an elevated risk of perioperative morbidity and mortality.


A qualitative study of the duty to care in communicable disease outbreaks.
Health care providers' (HCPs') duty to care during communicable disease outbreaks has resurfaced as an important and contentious topic. This renewed interest follows the re-emergence of communicable diseases, largely thought to have disappeared and therefore irrelevant to modern day practitioners. The 2003 SARS outbreak particularly presented propitious circumstances for reconsidering this issue. This study seeks to characterize the views of individuals on the nature and limits of this duty. The authors employed qualitative methods to gather lay and expert perspectives. Individual interviews were conducted with 67 participants consisting of HCPs, spiritual leaders, regulators, and members of the public from the greater Toronto area. Participants' views were analyzed and organized according to three main themes, constituting a framework that combines micro-, meso-, and macro-level structures and processes: the scope of obligations of HCPs, the roles of health care institutions, and the broader social context, respectively. Our data suggest that the duty to care must be placed in a wider context to include considerations that transcend individual provider obligations. It thus follows, based on our data, that the duty to care cannot be left to personal choice or an appeal to morality based on an ethic derived entirely from individual obligations. The micro-meso-macro analytical framework that we have developed can guide the articulation of accepted norms of duty to care during epidemics and the development of policy for public health crises. It can also enhance the focus of our current expectations of HCPs' duty during epidemics. This can be achieved by informing regulatory bodies, collaborating with policy makers and engaging the public.


A surgical perspective of ERAS guidelines in thoracic surgery.
Guidelines for enhanced recovery after surgery (ERAS) have recently been published for lung surgery. Although some of the recommendations are generic or focused on anesthetic and nursing care, other recommendations are more specific to a thoracic surgeon's practice. The present review concentrates on the surgical approach, optimal chest drain management, and the importance of early mobilization. Most lung cancer resections are still performed via an open thoracotomy approach. If a thoracotomy is to be used, a muscle-sparing approach may result in reduced pain and better postoperative function. Sparing of the intercostal bundle also reduces pain. There is now evidence that minimally invasive surgery for early lung cancer results in superior patient outcomes. Postoperatively, single chest tubes should be used without the routine application of external suction. Digital drainage systems are more reliable and may produce superior outcomes. Conservative chest drain removal policies are unnecessary and impair patient recovery. Early mobilization protocols should be instigated to reduce postoperative complications. The use of ERAS after lung surgery has the potential to improve patient outcomes. Although specific surgical elements are in the minority, thoracic surgeons should be involved in all aspects of perioperative care as part of the wider multidisciplinary team.


Subcellular fractionation of the longitudinal smooth muscle/myenteric plexus of dog ileum: dissociation of the distribution of two plasma membrane marker enzymes.
The distribution of plasma membrane markers, the sodium pump [evaluated as ouabain-sensitive, potassium-stimulated p-nitrophenyl phosphatase (K+-pNPPase)], [3H]saxitoxin binding, and 5'-AMPase, was studied in the subcellular fractions prepared from the homogenates of the longitudinal smooth muscle/myenteric plexus of dog ileum. The K+-pNPPase activity and [3H]-saxitoxin binding were found to be predominantly associated with the synaptosomal fraction as indicated by the high level of these activities in the crude synaptosomal fraction and by the copurification of K+-pNPPase and [3H]saxitoxin binding, but not 5'-AMPase, with several synaptosomal markers during the fractionation of the crude synaptosomal fraction on density gradients. In contrast to the K+-pNPPase activity and [3H]saxitoxin binding, the 5'-AMPase activity was found to be concentrated in the microsomal pellet. Further fractionation of microsomes on density gradient resulted in copurification of 5'-AMPase but not K+-pNPPase or [3H]saxitoxin binding, with other smooth muscle plasma membrane-bound enzymes, such as high-affinity Ca2+-ATPase, Mg2+-ATPase, and Ca2+-ATPase. It was concluded that in the longitudinal smooth muscle/myenteric plexus, the sodium pump activity is present in higher density in the neuronal plasma membranes whereas 5'-AMPase activity is concentrated in the smooth muscle plasma membranes.


Retinoic acid receptor beta induces terminal differentiation of squamous cell carcinoma lines in the absence of cyclin-dependent kinase inhibitor expression.
The chemotherapeutic agent and vitamin A metabolite retinoic acid (RA) has been used to treat many tumor types. The effects of RA are mediated by a family of ligand-dependent transcription factors, the retinoic acid receptors (RAR) and the retinoid X receptors. Alterations in RAR expression have been implicated in tumorigenesis. We showed previously that loss of RARbeta expression and target gene responsiveness to RA were consistent features of head and neck squamous cell carcinoma lines. To begin to elucidate the role of RARbeta in the malignant transformation of stratified squamous epithelia, we reexpressed RARbeta in squamous cell carcinoma (SCC) lines by stable transfection. SCC lines expressing RARbeta produced large numbers of flat cells with abundant cytoplasm that died and detached from the culture dish. These cells morphologically resembled the differentiated cells of normal stratified squamous epithelium in culture. These cells did not exhibit the characteristic DNA fragmentation pattern of apoptotic cells, nor did they label in a fluorescent apoptosis assay. Northern and Western blotting revealed induction of terminal differentiation markers in these cells. RARbeta produced alterations in levels of RA-responsive gene expression in these cells, and terminal differentiation occurred in the absence of detectable cyclin-dependent kinase inhibitor expression. We concluded that RARbeta expression induced terminal differentiation in SCC lines, suggesting a role for this transcription factor in the normal maturation of stratified squamous epithelium.


Electrooptic modulation methods for high sensitivity tunable diode laser spectroscopy.
A CdTe phase modulator and low power rf source have been used with Pb-salt tunable diode lasers operating near 8 microm to generate optical sidebands for high sensitivity absorption spectroscopy. Sweep averaged, first-derivative sample spectra of CH(4) were acquired by wideband phase sensitive detection of the electrooptically (EO) generated carrier-sideband beat signal. EO generated beat signals were also used to frequency lock the TDL to spectral lines. This eliminates low frequency diode jitter, and avoids the excess laser linewidth broadening that accompanies TDL current modulation frequency locking methods.


A material sparing test to predict punch sticking during formulation development.
A qualitative and material sparing test for punch sticking was developed as a predictive development screening tool for comparison of formulation strategies and processing methods to eliminate punch adhesion. To develop a predictive test for a formulation's potential to adhere to tablet punches during compression. Two proprietary compounds, A and B, with histories of punch sticking during manufacturing but not in development, were formulated with microcrystalline cellulose, lactose, croscarmellose sodium, and magnesium stearate. Matching placebos were also prepared by the same methods. Flat faced bevel edge (FFBE) punches of 1/2" diameter were compared with 1/4" SRC punches and 1/4" Flat Faced (FF) punches for adhesion potential using a Kilian rotary tablet press and a Huxley Bertram compaction simulator. Formulation accumulated on the punches after compression, which indicated potential for punch sticking that resulted in defective tablets with successive compactions. In all cases, only active formulations adhered to the FFBE and FF punches during tableting runs and with fewer compactions than the SRC punches. Those APIs were then reformulated and successfully manufactured using the predictive test results. Reducing the punch size to 1/4" permitted testing with 50 g of formulation, a ten-fold reduction in consumption compared to the 1/2" punches. Flat surfaced punches were shown to be more challenging to adhesive formulations than SRC punches. Adhesive formulations were reformulated and successfully manufactured without punch sticking.


Implementation of a Multifaceted Interactive Electrodiagnostic Medicine Workshop in a Physical Medicine and Rehabilitation Residency Program.
Electrodiagnostic medicine is a required component of Physical Medicine and Rehabilitation residency education, but limited resources exist to guide curriculum development. Our objective was to create a focused workshop to enhance our residency program's electrodiagnostic curriculum. We created two separate 1.5-day workshops, one basic and one advanced, for all residents. Each workshop included didactic sessions, case discussion, question and answer sessions, demonstrations, and hands-on participation with direct supervision and feedback. Presurveys and postsurveys were administered to evaluate the value of the workshops. We also assessed trends in electrodiagnostic self-assessment examination scores. Residents reported clinical electrodiagnostic rotations to be more valuable to their education than previous didactic sessions and independent learning. Self-reported knowledge of electrodiagnostic concepts, resident comfort level in planning, performing, and interpreting studies, and perceived value in independent learning of electrodiagnostic medicine improved after implementation of the workshops. There was a 7% improvement in the American Association of Neuromuscular and Electrodiagnostic Medicine electrodiagnostic self-assessment examination score compared with the previous year and a 15% improvement in the Physical Medicine and Rehabilitation self-assessment examination electrodiagnostic subscore compared with the previous 5 yrs. All participants recommended similar educational experience for other residents. This successful workshop may serve as a resource for other training programs.


Benign glandular cells in posthysterectomy vaginal smears: the incidence is higher than expected.
In this study, we have investigated the frequency and clinical significance of glandular cells in posthysterectomy vaginal smears. The slides of vaginal cuff smears of 290 patients were reviewed. The glandular cells were categorized into three groups: (1) squamous metaplastic-like cells; (2) columnar endocervical-like cells; and (3) small round cuboidal cells. Glandular cells were found in 39 (13%) of the 290 vaginal smears. Group 1 type cells were seen in 76% (n = 30), group 2 type cells in 38% (n = 15) and group 3 type cells in 47% (n = 19) of the smears. In 19 (48%) of the smears combination of two or three groups were seen. The presence of glandular cells showed a strong association with inflammation/repair as a background finding in the smears. No correlation could be found between the presence of atrophy and history of chemotherapy or radiotherapy. Apart from these there was a prominent increase in reporting benign glandular cells after the application of the current Bethesda 2001 reporting criteria in our laboratory. As a result our study showed that the finding of glandular cells in posthysterectomy vaginal smears is more frequent than expected and most of them could be related to inflammatory and regenerative processes in the absence of a clearly identified underlying cause.


Efficacy of a Novel Optimized Pulsed Light Source (MaxG) for the Treatment of Facial Vascular Lesions.
Facial vascular malformations can cause a number of functional problems, including difficulties in breathing, eating, speech, and mobility. Psychological problems can also arise due to the possible unpleasant appearance of such lesions. Further, these lesions can lead to a number of complications, including pain, ulceration, infection, and significant bleeding. Many treatments have been proposed in the literature. Laser therapy (and its related treatments by non-coherent light sources) is now considered the gold standard in the treatment of the majority of vascular lesions. Here, we present our experience with a novel optimized pulsed light source for the treatment of vascular anomalies. In this prospective study, we evaluate the clinical outcomes of 30 patients treated with this method. Our results confirm the efficacy and safety of this treatment for facial vascular lesions. Based on our experience and results, we believe this device could be considered as both an alternative monotherapy and a useful adjunctive to the already existing laser instruments.


Parathyroid hormone-related protein and calcium homeostasis in lactating mice.
Parathyroid hormone (PTH)-related protein has been shown to be a factor responsible for hypercalcemia of malignancy. Recent studies have shown the presence of mRNA for PTH-related protein in lactating breast tissue, suggesting a physiological role for this peptide during lactation. In the present study, we evaluated the effect of neutralization of PTH-related protein activity in lactating mice (by passive immunization) on various parameters of maternal and neonatal calcium homeostasis. PTH-related protein bioactivity, as tested in the adenylate cyclase assay, was present in mouse milk, and this activity was completely neutralized by the antisera used in the present study. In lactating mice, the effects of injection of PTH-related protein antisera on maternal serum calcium concentrations, milk calcium and phosphorus concentration, pup growth, dam femur calcium content, and pup calcium content were similar to those of the injection of normal rabbit serum. Therefore, maternal PTH-related protein does not appear to have a role in calcium homeostasis during lactation.


New opportunities for enhancing oral health: moving toward the 1990 objectives for the nation.
In July 1983, the Assistant Secretary for Health reviewed progress toward achievement of a dozen national objectives in fluoridation and dental health. These 12 objectives, classified under the categories of improved oral health status, reduced risk factors, increased public and professional awareness, improved services and protection, and improved surveillance-evaluation systems, hold promise for improved oral health in this country. It is noteworthy that the objective that 40 percent of 9-year-old children be caries-free in their permanent dentition has been accomplished (51 percent of 9-year-olds were caries-free according to a 1979-80 National Institute of Dental Research study). Still, dental caries is highly prevalent among teenaged children, and gingival and periodontal conditions are highly prevalent among children and adults. A number of highly effective methods are available for preventing dental decay in children as well as adults; they include community or school water fluoridation, the use of multiple forms of supplemental fluorides, avoidance of frequent consumption of foods that are high in sugar content, and the use of adhesive pit and fissure sealants. Personal use and professional provision of these methods in appropriate combinations can contribute significantly to future improvements in oral health. Meticulous personal oral hygiene practices combined with periodic professional care are the currently available means of protecting periodontal health. Through increased collaboration among various governmental, academic, and corporate entities, as well as active participation by individuals, the achievement of a number of these objectives becomes feasible to the benefit of national productivity, health care financing, and the quality of life for Americans.


Anomalous small angle x-ray scattering simulations: proof of concept for distance measurements for nanoparticle-labelled biomacromolecules in solution.
Anomalous small angle X-ray scattering can in principle be used to determine distances between metal label species on biological molecules. Previous experimental studies in the past were unable to distinguish the label-label scattering contribution from that of the molecule, because of the use of atomic labels; these labels contribute only a small proportion of the total scattering signal. However, with the development of nanocrystal labels (of 50-100 atoms) there is the possibility for a renewed attempt at applying anomalous small angle X-ray scattering for distance measurement. This is because the contribution to the scattered signal is necessarily considerably stronger than for atomic labels. Here we demonstrate through simulations, the feasibility of the technique to determine the end-to-end distances of labelled nucleic acid molecules as well as other internal distances mimicking a labelled DNA binding protein if the labels are dissimilar metal nanocrystals. Of crucial importance is the ratio of mass of the nanocrystals to that of the labelled macromolecule, as well as the level of statistical errors in the scattering intensity measurements. The mathematics behind the distance determination process is presented, along with a fitting routine than incorporates maximum entropy regularisation.


Gastric distension-induced c-fos expression in catecholaminergic neurons of rat dorsal vagal complex.
Functionally specific vagal afferents were stimulated by gastric balloon distension in unanesthetized rats, followed by double c-fos/dopamine beta-hydroxylase (DBH) immunocytochemistry, to identify second-order neurons in the dorsal vagal complex. Continuous and repeated phasic distension with similar volumes produced similar numbers and patterns of c-fos expression, with most of the activated neurons in the medial and commissural nucleus of the solitary tract (NTS) and dorsal motor nucleus (DMNX). Larger distension activated significantly more neurons in all responsive areas but there was no differential effect. In most NTS subnuclei and the DMNX, a small (3-5%) proportion of gastric distension-activated neurons was DBH-immunoreactive (DBH-IR), and this proportion did not significantly change with type of distension. With continuous and repeated small distensions, 10-12% and, with the large distension, 22-30% of all DBH-IR neurons expressed c-fos. The results suggest a large degree of convergence between rapidly adapting mucosal receptors and slowly adapting tension receptors, but not between low- and high-threshold tension receptors, and a relatively minor role of catecholaminergic second-order neurons in the dissemination of distension signals in the brain.