Development of DexAide right ventricular assist device: update II.
The DexAide right ventricular assist device (RVAD) is a magnetically and hydrodynamically levitated implantable centrifugal pump. Recent progress includes 1) redesign of the inflow/outflow conduits, which yielded two successful 3-month experiments, 2) development of alternative journal bearing materials, and 3) completion of an 18-month duration of in vitro endurance testing. Verification testing of the RVAD electronics has been completed, and a prototype biventricular assist device (BVAD) system has been tested. Acute DexAide/CorAide BVAD implantations via median sternotomy in two calves documented BVAD control algorithms and anatomical fit. A drug-induced chronic calf heart failure model, currently under development in our laboratory, resulted in a successful BVAD implantation in a calf with heart failure. Our future plans are to complete in vitro and in vivo validation of alternative bearing materials, perform preclinical DexAide in vivo and in vitro reliability studies, and obtain Food and Drug Administration (FDA) approval for an Investigational Device Exemption to conduct a clinical pilot study. In conclusion, two successful 3 month in vivo experiments and an 18-month in vitro endurance test were completed. After final bearing material selection, the DexAide design will be "frozen" so that preclinical systems can be manufactured. BVAD experiments using a chronic heart failure model are in progress.


Characterization of mannose 6-phosphate receptors (MPRs) from opossum liver: opossum cation-independent MPR binds insulin-like growth factor-II.
Bovine, human, and rat cation-independent mannose 6-phosphate receptors (CI-MPRs) are capable of binding both mannose 6-phosphate and insulin-like growth factor-II (IGF-II). However, the receptor isolated from either chicken or frog lacks the high affinity IGF-II-binding site. To determine whether CI-MPRs isolated from a species that is closely related to placental mammals can bind IGF-II, the MPRs were purified from a marsupial, the American opossum (Didelphis virginiana), by phosphomannan-Sepharose affinity chromatography and then tested for their ability to bind IGF-II. Opossum liver expressed both the CI-MPR and the cation-dependent MPR (CD-MPR). Both receptors contained Asn-linked oligosaccharides. In contrast to CD-MPRs isolated from other species, the opossum CD-MPR displayed heterogeneity with respect to the number of Asn-linked oligosaccharide chains it contains. The CI-MPR isolated from opossum liver, like the CI-MPR from bovine liver, bound iodinated human recombinant IGF-II. However, Scatchard analysis revealed that the opossum CI-MPR bound IGF-II with a lower affinity (Kd = 14.5 nM) than the bovine receptor (Kd = 0.2 nM). The addition of excess IGF-II, but not IGF-I or insulin, inhibited binding to [125I]IGF-II, indicating that the opossum CI-MPR exhibits specificity for IGF-II. These results suggest that the emergence of a high affinity IGF-II-binding site in the CI-MPR occurred in evolution before the divergence of marsupials and placental mammals from their last common ancestor.


Production of a Mouse Monoclonal Antibody Against Mortalin by Whole Cell Immunization.
Pancreatic carcinoma is the fourth leading cause of cancer death and is characterized by early invasion and metastasis. Advances in molecular biology directed new strategies in targeted therapy using monoclonal antibodies. To identify new biomarkers, we generated a panel of monoclonal antibodies against the newly established cell line, Faraz-ICR, from a patient with acinar cell carcinoma. After immunization of BALB/c female mice with Faraz-ICR cell line and fusion of splenocytes with SP2/0 myeloma cell line, high reactive hybridoma producing antibodies to Faraz-ICR were detected using enzyme-linked immunosorbent assay, immunofluorescence staining and flow cytometry. Western blot and two-dimensional immunoblot were used for further characterization of the targets antibodies. Among high reactive clones, the reactivity of 1C11 clone was assessed with other epithelial tumors. The isotype of the antibody was revealed to be IgM, and the antibody reacted to a protein with a molecular weight of about 70 kDa in Western blot analysis. To further characterization of the target antigen, immunoproteome of the Faraz-ICR cell line was performed. By liquid chromatography-mass spectrometry (LC-MS) analysis, we identified that the target of 1C11 clone was HSP70. In conclusion, pancreatic cancer is a fatal malignancy with no reliable biomarker for early screening and diagnosis. In this study, by establishing a pancreatic cell line, a panel of monoclonal antibodies was generated aiming to explore specific or associated cancer targets. We then introduced 1C11 monoclonal antibody that can specifically recognize mortalin as a main tumor marker and may serve as a new tool for diagnostic kit and therapeutic strategies targeting this molecule.


Amphibian parasites of the Order Dermocystida (Ichthyosporea): current knowledge, taxonomic review and new records from Brazil.
Amphibian parasites of the Order Dermocystida (Ichthyosporea) are widespread pathogens known mainly from Europe and North America, which cause primarily a disease of skin and subcutaneous tissue in their hosts. The taxonomy of these organisms has been problematic given their conserved morphology, similar clinical disease and pathology. Currently recognized taxa belong to the three closely related genera, Amphibiocystidium, Amphibiothecum, and Rhinosporidium, whereas species of Dermocystidium and Sphaerothecum destruens include fish parasites. Here, we review the taxonomy of Dermocystida based on a molecular phylogenetic analysis, principally of amphibian parasites, including DNA sequences obtained from amphibian hosts collected in the central-eastern region of South America. A new taxonomic arrangement is proposed, which includes the designation of type material for Dermocystidium pusula, synonymization of Amphibiothecum with Dermocystidium, and the restriction of Amphibiocystidium to its type species A. ranae. We also review the taxonomic status of Dermosporidium hylarum until the present work included in the synonymy of the human and animal pathogen R. seeberi, and considered herein as a valid taxon, however. In addition, a new species of Sphaerothecum parasitic to amphibians is described, being the first record of this genus in the southern hemisphere and in an amphibian host.


System for measuring planar lipid bilayer properties.
We present a system for measuring planar lipid bilayer properties. The system is composed of a control unit, an output stage, an LCR meter, pumps for filling reservoirs, a bath with temperature regulation and a measurement chamber with four electrodes. The planar lipid bilayer is automatically formed using a folding method on apertures of different sizes. The automatization is assured by two syringes, which are clamped in actuators. Actuators are driven and controlled by a control unit via RS-232 communication. The temperature of the planar lipid bilayer can be regulated between 15 and 55 °C. The regulation is assured by insertion of the measurement chamber into the temperature-regulated bath. Different shapes of voltage- or current-clamp signals can be applied to the planar lipid bilayer. By measuring the response of the planar lipid bilayer to the applied signal, the capacitance and breakdown voltage of the planar lipid bilayer can be determined. The cutoff frequencies of the system output stage for voltage- and current-clamp methods are 11 and 17 kHz, respectively.


[Therapy of angina pectoris].
Treatment of angina pectoris should follow the clinical course: Stable angina is most often responding to nitropreparations, betablockers or calciumantagonists; differentialtherapy should consider the basic state of the patient: age, heart-rate, hypo- or hypertension, cardiac failure or asthma bronchiale etc. Unstable angina needs more attention and should be transferred to a coronary care unit. Analgetics, sedatives, oxygen and nitrates should be applied under hemodynamic monitoring. Only in rare cases with cardiac failure or with rapid atrial fibrillation glycosides will be necessary. In extreme bradiacardias a pacemaker can be helpful. Captopril might be a new substance in nonresponders.


Mechanical and current responses to neurotensin in the smooth muscle of guinea-pig intestine.
1. The effects of neurotensin (NT) on the mechanical activity of the smooth muscle and membrane currents of enzymatically-dispersed single smooth muscle cells of guinea-pig intestine were investigated by the isometric tension recording method and the whole-cell patch clamp technique, respectively. 2. NT (0.3-670 nM) produced an initial relaxation followed by a contraction in segment preparations of the jejunum and ileum. Atropine (0.2 microM) abolished the contraction. In 30% of ileal segments, NT produced a slowly-developed contraction in the presence of atropine. The atropine-resistant contraction was insensitive to tetrodotoxin (TTX, 0.2 microM). The relaxant effect of NT was unaffected by TTX (0.2 microM) and guanethidine (2 microM), but markedly reduced or abolished by apamin (67 nM). 3. In segment preparations of the colon and rectum, NT produced a biphasic response, similar to that in the small intestine, or a triphasic one (relaxation, contraction and relaxation). The contractile effect of NT was affected neither by atropine (0.2 microM) nor by TTX (0.2 microM). The first relaxation in response to NT was similar in pharmacological properties to that in the small intestine. 4. Responses to NT in longitudinal muscle strips were similar to those in segment preparations. The taenia caecum responded to NT with a contraction alone and the effect was unaffected by atropine (0.2 microM) and TTX (0.2 microM). 5. NT had little or no effect on the mechanical activity in the circular muscle of the small intestine. In the circular muscle of the large intestine, NT had a weak inhibitory effect on the spontaneous activity which was followed by a small rise in muscle tone. Apamin (67 nM) converted the biphasic response to a contraction. 6. In cells dialysed with a KCl-based solution under voltage clamp at 0 mV, NT (5 microM) produced a brief, outward current in a very small fraction of cells (4 out of 40 cells). When cells were dialysed with a CsCl-based solution under voltage clamp at -50 or -60 mV, no current response to NT (5 microM) was observed in all 37 cells, but NT increased inward Ca2+ currents evoked upon depolarization. 7. From these results, it appears that there is a regional difference in the mode of action of NT to contract and relax the smooth muscle of guinea-pig intestine, and NT acts on its receptors on the smooth muscle to enhance activation of voltage-dependent Ca2+ channels, which underlies the slow contraction in the longitudinal muscle of the intestine.


Measurement-device-independent quantum key distribution over 200 km.
Measurement-device-independent quantum key distribution (MDIQKD) protocol is immune to all attacks on detection and guarantees the information-theoretical security even with imperfect single-photon detectors. Recently, several proof-of-principle demonstrations of MDIQKD have been achieved. Those experiments, although novel, are implemented through limited distance with a key rate less than 0.1 bit/s. Here, by developing a 75 MHz clock rate fully automatic and highly stable system and superconducting nanowire single-photon detectors with detection efficiencies of more than 40%, we extend the secure transmission distance of MDIQKD to 200 km and achieve a secure key rate 3 orders of magnitude higher. These results pave the way towards a quantum network with measurement-device-independent security.


Use of natural remedies to treat oral diseases among female patients in Al Madinah, western Saudi Arabia.
Background The study aimed to investigate natural remedies use (NRU) for oral diseases among female patients. Methods Dental patients attending the female section of Taibah University Dental Hospital were invited to complete an online questionnaire which explored demographic data, and pattern of NRU for oral diseases. Results A total of 540 patients participated in the survey. A total of 232 patients (43%) used natural remedies to treat oral diseases. The most commonly cited reasons for NRU were financial reasons (n=79, 34.1%). The practice had a statistically significant association with older participants (p=0.003), and those working as employees (p=0.031). Most of the natural remedies used (n=17) were herbal, with cloves (71.1%) and miswak (26.7%) being the most frequently reported. Acute dental pain (n=157, 67.7%) and halitosis (n= 51, 22%) were the most frequently cited oral conditions associated with NRU. A percentage of 87.1% of patients using natural remedies (n=202) perceived improvement of symptoms upon use of natural remedies, however, 2.2% (n=5) reported complications. Conclusions Substantial proportion of patients use natural remedies to treat their oral disease. Patients were mostly satisfied with natural remedies in terms of effectiveness and lack of complications.


Radioisotope scanning in inflammatory muscle disease.
Fourteen whole-body rectilinear bone scans using technetium 99m-polyphosphate were done in nine patients with well-documented inflammatory myopathy (either polymyositis or dermatomyositis). In all nine patients, the scans showed evidence of increased muscle labeling. Muscle uptake was markedly increased in one patient, moderately increased in two patients, and minimally increased in six patients. The degree of muscle labeling correlated with the severity of the muscle weakness at the time the scan was done. In four patients, who received high-dose corticosteroid treatment, muscle uptake was decreased following therapy. These findings suggest that radioisotope scanning may be useful in the diagnosis and management of patients with inflammatory muscle diseases.


Autocrine regulation of neural crest cell development by steel factor.
Steel factor (SLF) and its cognate receptor, c-kit, have been implicated in the generation of melanocytes from migrating neural crest (NC) cells during early vertebrate embryogenesis. However, the source of SLF in the early avian embryo and its precise role in melanogenesis are unclear. We report here that NC cells themselves express and release SLF protein, which in turn acts as an autocrine factor to induce melanogenesis in nearby NC cells. These results indicate that NC cell subpopulations play an active role in the determination of their cell fate and suggest a different developmental role for the embryonic microenvironment than what has been previously proposed.


Chaotic dynamics of a swirling flame front instability generated by a change in gravitational orientation.
We have intensively examined the dynamic behavior of flame front instability in a lean swirling premixed flame generated by a change in gravitational orientation [H. Gotoda, T. Miyano, and I. G. Shepherd, Phys. Rev. E 81, 026211 (2010)PLEEE81539-375510.1103/PhysRevE.81.026211] from the viewpoints of complex networks, symbolic dynamics, and statistical complexity. Here, we considered the permutation entropy in combination with the surrogate data method, the permutation spectrum test, and the multiscale complexity-entropy causality plane incorporating a scale-dependent approach, none of which have been considered in the study of flame front instabilities. Our results clearly show the possible presence of chaos in flame front dynamics induced by the coupling of swirl-buoyancy interaction in inverted gravity. The flame front dynamics also possesses a scale-free structure, which is reasonably shown by the probability distribution of the degree in ε-recurrence networks.


NACCHO GP Network--enhancing communication in Aboriginal health.
Communication between general practitioners, information sharing, and GP support are important issues, especially for those in rural and remote areas. Internet based links can facilitate enhanced communication. This article describes the new internet based communication forum for GPs working, or interested in the health of, Aboriginal and Torres Strait Islander peoples. The NACCHO GP Network is the first on-line service to provide focussed information for GPs in the Aboriginal community controlled health sector. It also provides a place for GPs, locums and general practice registrars to raise issues and share ideas and information on Aboriginal health practice. The NACCHO GP Network has the potential to reduce isolation and address the support needs of GPs, and enhance the interaction of the Aboriginal community controlled sector with general practice organisations.


Disparate folding and stability of the ankylosing spondylitis-associated HLA-B*1403 and B*2705 proteins.
To investigate the folding, assembly, maturation, and stability of HLA-B*1402 and B*1403, which differ by 1 amino acid change and are differentially associated with ankylosing spondylitis (AS), and to compare these features with those of B*2705. Stable transfectants expressing B*1402, B*1403, and B*2705 were used. Folding rates were estimated from the ratio of unfolded heavy chains to folded heavy chains that had been immunoprecipitated with specific antibodies in pulse-chase experiments. Heavy chain misfolding was measured as the half-life of endoglycosidase H (Endo H)-sensitive beta2-microglobulin-free heavy chains. Maturation/export rates were measured by acquisition of Endo H resistance. Association with calnexin or tapasin was analyzed by coprecipitation with chaperone-specific antibodies, and surface expression was estimated by flow cytometry. Thermostability of HLA-peptide complexes was assessed by immunoprecipitation after incubation at various temperatures. Heavy chain expression was quantified by Western blotting. The folding rates of B*1402 and B*1403 were similar, and both were faster and more efficient than B*2705, but some unfolded heavy chains from both B14 subtypes remained in the endoplasmic reticulum (ER) with a long half-life. The export rates of B*1402 and B*1403 were slow, and the heterodimers partially dissociated after exiting the ER, as revealed by significant amounts of Endo H-resistant and surface-expressed free heavy chains. Both interaction with tapasin and thermostability were higher for B*2705 than for B*1402 and higher for B*1402 than for B*1403, suggesting that the repertoires of the B*1402-bound peptide and especially the B*1403-bound peptide were less optimized than that of B*2705. Our results indicate that the folding, maturation, and stability of B*1403 differ more from B*2705 than from B*1402. Thus, these features cannot account for the fact that only the 2 former allotypes are associated with AS.


An anatomical, histochemical, and autoradiographic study of the ever-growing molar dentition of Microtus with comments on the role of structure in growth and eruption.
An analysis of the microanatomy of the molar dentition of Microtus utilizing histological, histochemical and autoradiographic techniques reveals a complex architecture with distinctive morphogenic mechanisms which respond to the functional requirements of the organism. These mechanisms include; the maintenance of continued growth and eruption of the molars to compensate for continued hard tissue loss from wear at the occlusal surface of the crown throughout the entire lifespan of the orgainism and a positive feedback repair mechanism to protect the growth systems from the potential destruction this normal occlusal wear could initiate. An awareness and understanding of these phenomena is of significant value for interpreting palentological specimens and formulating a theoretical model for interpreting the evolution of Microtine molar dentitions.


Interleukin-4 Induces CpG Site-Specific Demethylation of the Pendrin Promoter in Primary Human Bronchial Epithelial Cells.
Pendrin is upregulated in bronchial epithelial cells following IL-4 stimulation via binding of STAT6 to an N4 GAS motif. Basal CpG methylation of the pendrin promoter is cell-specific. We studied if a correlation exists between IL-4 sensitivity and the CpG methylation status of the pendrin promoter in human bronchial epithelial cell models. Real-time PCR and pyrosequencing were used to respectively quantify pendrin mRNA levels and methylation of pendrin promoter, with and without IL-4 stimulation, in healthy and diseased primary HBE cells, as well as NCI-H292 cells. Increases in pendrin mRNA after IL-4 stimulation was more robust in NCI-H292 cells than in primary cells. The amount of gDNA methylated varied greatly between the cell types. In particular, CpG site 90 located near the N4 GAS motif was highly methylated in the primary cells. An additional CpG site (90bis), created by a SNP, was found only in the primary cells. IL-4 stimulation resulted in dramatic demethylation of CpG sites 90 and 90bis in the primary cells. IL-4 induces demethylation of specific CpG sites within the pendrin promoter. These epigenetic alterations are cell type specific, and may in part dictate pendrin mRNA transcription.


Influence of βS-Globin Haplotypes and Hydroxyurea on Arginase I Levels in Sickle Cell Disease.
Introduction. Sickle cell disease (SCD) is characterized by hemoglobin S homozygosity, leading to hemolysis and vasoocclusion. The hemolysis releases arginase I, an enzyme that decreases the bioavailability of nitric oxide, worsening the symptoms. The different SCD haplotypes are related to clinical symptoms and varied hemoglobin F (HbF) concentration. The aim of this study was to evaluate the impact of the βS gene haplotypes and HbF concentration on arginase I levels in SCD patients. Methods. Fifty SCD adult patients were enrolled in the study and 20 blood donors composed the control group. Arginase I was measured by ELISA. The βS haplotypes were identified by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Statistical analyses were performed with GraphPad Prism program and the significance level was p < 0.05. Results. Significant increase was observed in the arginase I levels in SCD patients compared to the control group (p < 0.0001). The comparison between the levels of arginase I in three haplotypes groups showed a difference between the Bantu/Bantu × Bantu/Benin groups; Bantu/Bantu × Benin/Benin, independent of HU dosage. An inverse correlation with the arginase I levels and HbF concentration was observed. Conclusion. The results support the hypothesis that arginase I is associated with HbF concentration, also measured indirectly by the association with haplotypes.


Multi-omics Analysis of Periodontal Pocket Microbial Communities Pre- and Posttreatment.
Periodontitis is a polymicrobial infectious disease that causes breakdown of the periodontal ligament and alveolar bone. We employed a meta-omics approach that included microbial 16S rRNA amplicon sequencing, shotgun metagenomics, and tandem mass spectrometry to analyze sub- and supragingival biofilms in adults with chronic periodontitis pre- and posttreatment with 0.25% sodium hypochlorite. Microbial samples were collected with periodontal curettes from 3- to 12-mm-deep periodontal pockets at the baseline and at 2 weeks and 3 months. All data types showed high interpersonal variability, and there was a significant correlation between phylogenetic diversity and pocket depth at the baseline and a strong correlation (rho = 0.21; P = 0.008) between metabolite diversity and maximum pocket depth (MPD). Analysis of subgingival baseline samples (16S rRNA and shotgun metagenomics) found positive correlations between abundances of particular bacterial genera and MPD, including Porphyromonas, Treponema, Tannerella, and Desulfovibrio species and unknown taxon SHD-231. At 2 weeks posttreatment, we observed an almost complete turnover in the bacterial genera (16S rRNA) and species (shotgun metagenomics) correlated with MPD. Among the metabolites detected, the medians of the 20 most abundant metabolites were significantly correlated with MPD pre- and posttreatment. Finally, tests of periodontal biofilm community instability found markedly higher taxonomic instability in patients who did not improve posttreatment than in patients who did improve (UniFrac distances; t = -3.59; P = 0.002). Interestingly, the opposite pattern occurred in the metabolic profiles (Bray-Curtis; t = 2.42; P = 0.02). Our results suggested that multi-omics approaches, and metabolomics analysis in particular, could enhance treatment prediction and reveal patients most likely to improve posttreatment. IMPORTANCE Periodontal disease affects the majority of adults worldwide and has been linked to numerous systemic diseases. Despite decades of research, the reasons for the substantial differences among periodontitis patients in disease incidence, progressivity, and response to treatment remain poorly understood. While deep sequencing of oral bacterial communities has greatly expanded our comprehension of the microbial diversity of periodontal disease and identified associations with healthy and disease states, predicting treatment outcomes remains elusive. Our results suggest that combining multiple omics approaches enhances the ability to differentiate among disease states and determine differential effects of treatment, particularly with the addition of metabolomic information. Furthermore, multi-omics analysis of biofilm community instability indicated that these approaches provide new tools for investigating the ecological dynamics underlying the progressive periodontal disease process.


Draft Genome Sequences of Three O157 Enteropathogenic Escherichia coli Isolates.
We report the draft genome sequences of three enteropathogenic Escherichia coli (EPEC) isolates that display the O157 serogroup but do not have the Shiga toxin genes (stx), which are characteristic of O157 enterohemorrhagic E. coli (EHEC). E. coli strain RN587/1 has the O157:H8 serotype and possesses the EAF plasmid characteristic of typical EPEC (J. B. Kaper, J. P. Nataro, and H. L. Mobley, Nat. Rev. Microbiol. 2:123-140, 2004). The other two isolates, strains C844-97 and C639-08, are both O157:H45 and possess the locus of enterocyte effacement (LEE) pathogenicity island; however, they do not contain the EAF plasmid or the stx-carrying phage.


The role of medical comorbidity in outcome of major depression in primary care: the PROSPECT study.
The authors described the influence of specific medical conditions on clinical remission and response of major depression (MDD) in a clinical trial evaluating a care-management intervention among older primary-care patients. Adults age 60 years and older were randomly selected and screened for depression. Participants were randomly assigned to Usual Care or to an Intervention with a depression care-manager offering algorithm-based care for MDD. In all, 324 adults meeting criteria for MDD were included in these analyses. Remission and response was defined by a score on the Hamilton Rating Scale for Depression <10 and by a decrease from baseline of > or =50%, respectively. Medical comorbidity was ascertained through self-report. Cognitive impairment was defined by a score <24 on the Mini-Mental State Exam (MMSE). In Usual Care, rates of remission were faster in persons who reported atrial fibrillation (AF) than in persons who did not report AF and slower in persons who reported chronic pulmonary disease than in persons who did not report chronic pulmonary disease; rates of response were less stable in persons with MMSE <24 than in those with MMSE > or =24. In the Intervention condition, none of the specific chronic medical conditions were significantly associated with outcomes for MDD. Because disease-specific findings were observed in persons who received Usual Care but not in persons who received more intensive treatment in the Intervention condition, our results suggest that the association of medical comorbidity and treatment outcomes for MDD may be determined by the intensity of treatment for depression.


Degenerate optical parametric amplifier delivering sub 30 fs pulses with 2GW peak power.
Degenerated optical parametric amplification (OPA) is a well known technique to achieve broadband amplification necessary to generate ultrashort pulses. Here we present a parametric amplifier pumped by the frequency doubled output of a state-of-the-art fiber chirped pulse amplification system (FCPA) delivering mJ pulse energy at 30 kHz repetition rate and 650 fs pulse duration. The parametric amplifier and the FCPA system are both seeded by the same Yb:KGW oscillator. Additional spectral broadening of the OPA seed provides enough bandwidth for the generation of ultrashort pulses. After amplification in two 1mm BBO crystals a pulse energy of 90 microJ is yielded at 30 kHz. Subsequent compression with a sequence of chirped mirrors shortens the pulses to 29 fs while the pulse energy is as high as 81 microJ resulting in 2GW of peak power.


Simulation of the kinetics of neuromuscular block: implications for speed of onset.
The onset time for paralysis varies 3-fold among nondepolarizing muscle relaxants. Possible explanations include: (a) pharmacokinetic differences among drugs and (b) buffering of drug molecules by acetylcholine receptors as they diffuse into the neuromuscular junction. Although some pharmacokinetic models consider buffered diffusion, these models do not account for either the high density of receptors or synapse geometry. Here, I used computer simulations to calculate the kinetics of buffered diffusion. The goal was to determine the conditions under which buffered diffusion could account for differences in onset time among nondepolarizing muscle relaxants. Monte Carlo simulation was used along with a realistic 3-dimensional model of the rat neuromuscular junction. Simulations determined the time dependence of the number of drug-bound receptors. A 1000-fold range of drug potency was examined. In some simulations, the drug concentration outside the junction was changed instantaneously. In other simulations, the concentration changed according to predictions of pharmacokinetic models assuming time-dependent changes in plasma drug concentration. The rate constant for equilibration of drug between plasma and muscle, keo, was varied between 0.15 and 0.6 min(-1). Twitch amplitude was calculated from receptor occupancy assuming a high safety margin for neuromuscular transmission. Some simulations used a synaptic model with an increased nerve-muscle contact width. Simulations with instantaneous changes in drug concentration at the synapse, indicated that the time to 50% twitch depression (onset time) was 0.1 to 30 seconds and was proportional to drug potency. This corresponds to iontophoretic application of drug to isolated neuromuscular junctions, but is too fast to explain onset times in humans. When pharmacokinetic models were used to calculate the drug concentration outside the synapse, buffered diffusion increased onset times of potent drugs (drugs for which the effective concentration at 50% twitch height is <600 nM). Simulations using keo = 0.6 min(-1) and a model with a 2- to 3-fold wider nerve-muscle contact width indicated that buffered diffusion could account for the differences in clinical onset times among the nondepolarizing muscle relaxants. Monte Carlo simulation provides a biophysically appropriate way to incorporate buffered diffusion into pharmacokinetic modeling. The simulations indicated that buffered diffusion could account for differences in onset time among drugs. However, a better understanding of the geometry of the human neuromuscular junction is needed before the magnitude of the effect of buffered diffusion can be quantified.


The monoclonal nature of lymphocytes in multiple myeloma. Effects of therapy.
Despite the increased amounts of serum immumoglobulins (Ig) in multiple myeloma, the percentage of bone marrow-dependent (B) cells carrying surface Ig in the peripheral blood compartment were decreased, when compared with that in control patients with polyclonal gammopathy or with that in normals (5 plus or minus 2% in patients with multiple myeloma and 19 plus or minus 5% or 22 plus or minus 7% in controls). Most circulating B cells in patients with multiple myeloma were shown to carry individually specific idiotypic Ig. The monoclonal idiotypic nature of the surface Ig was shown by using antiserums raised against the monoclonal Ig. The idiotypic antiserum was capable of actively binding to and stimulating the idiotypic lymphocytes to divide in vitro. Commercial anti-Ig antiserums raised against normal Ig did not bind to the myeloma lymphocytes. After chemotherapy, lymphocytes carrying the idiotypic Ig decreased in numbers, and this correlated with other variables of clinical improvements. Its role in the pathogenesis of myeloma and its importance for diagnosis and follow-up of patients with myeloma are discussed.


Biotin-tagged proteins: Reagents for efficient ELISA-based serodiagnosis and phage display-based affinity selection.
The high-affinity interaction between biotin and streptavidin has opened avenues for using recombinant proteins with site-specific biotinylation to achieve efficient and directional immobilization. The site-specific biotinylation of proteins carrying a 15 amino acid long Biotin Acceptor Peptide tag (BAP; also known as AviTag) is effected on a specific lysine either by co-expressing the E. coli BirA enzyme in vivo or by using purified recombinant E. coli BirA enzyme in the presence of ATP and biotin in vitro. In this paper, we have designed a T7 promoter-lac operator-based expression vector for rapid and efficient cloning, and high-level cytosolic expression of proteins carrying a C-terminal BAP tag in E. coli with TEV protease cleavable N-terminal deca-histidine tag, useful for initial purification. Furthermore, a robust three-step purification pipeline integrated with well-optimized protocols for TEV protease-based H10 tag removal, and recombinant BirA enzyme-based site-specific in vitro biotinylation is described to obtain highly pure biotinylated proteins. Most importantly, the paper demonstrates superior sensitivities in indirect ELISA with directional and efficient immobilization of biotin-tagged proteins on streptavidin-coated surfaces in comparison to passive immobilization. The use of biotin-tagged proteins through specific immobilization also allows more efficient selection of binders from a phage-displayed naïve antibody library. In addition, for both these applications, specific immobilization requires much less amount of protein as compared to passive immobilization and can be easily multiplexed. The simplified strategy described here for the production of highly pure biotin-tagged proteins will find use in numerous applications, including those, which may require immobilization of multiple proteins simultaneously on a solid surface.


Polyamine depletion prevents camptothecin-induced apoptosis by inhibiting the release of cytochrome c.
We have shown previously that depletion of polyamines delays apoptosis induced by camptothecin in rat intestinal epithelial cells (IEC-6). Mitochondria play an important role in the regulation of apoptosis in mammalian cells because apoptotic signals induce mitochondria to release cytochrome c. The latter interacts with Apaf-1 to activate caspase-9, which in turn activates downstream caspase-3. Bcl-2 family proteins are involved in the regulation of cytochrome c release from mitochondria. In this study, we examined the effects of polyamine depletion on the activation of the caspase cascade, release of cytochrome c from mitochondria, and expression and translocation of Bcl-2 family proteins. We inhibited ornithine decarboxylase, the first rate-limiting enzyme in polyamine synthesis, with alpha-difluoromethylornithine (DFMO) to deplete cells of polyamines. Depletion of polyamines prevented camptothecin-induced release of cytochrome c from mitochondria and decreased the activity of caspase-9 and caspase-3. The mitochondrial membrane potential was not disrupted when cytochrome c was released. Depletion of polyamines decreased translocation of Bax to mitochondria during apoptosis. The expression of antiapoptotic proteins Bcl-x(L) and Bcl-2 was increased in DFMO-treated cells. Caspase-8 activity and cleavage of Bid were decreased in cells depleted of polyamines. These results suggest that polyamine depletion prevents IEC-6 cells from apoptosis by preventing the translocation of Bax to mitochondria, thus preventing the release of cytochrome c.


Evaluation of Cytotoxicity and Cell Death Induced In Vitro by Saxitoxin in Mammalian Cells.
Since the cyanotoxin saxitoxin (STX) is a neurotoxin and induces ecological changes in aquatic environments, a potential risk to public and environmental health exists. However, data on STX-mediated cytotoxic and genotoxic effects are still scare. In order to gain a better understanding of the effects of this toxin, the cytotoxic and genotoxic potential of STX was examined in two mammalian cell lines. Neuro 2A (N2A), a neuroblastoma mouse cell line, and Vero cell line, derived from Vero green monkey kidney cells, were exposed to several concentrations of STX ranging from 0.5 to 64 nM to determine cell viability, induction of apoptosis (DNA fragmentation assay), and formation of micronuclei (MN) (cytokinesis-block micronucleus assay; CBMN) following 24 h of incubation. The half maximal effective concentration (EC50) values for STX calculated in cell viability tests were 1.01 nM for N2A and 0.82 nM for Vero cells. With increasing STX concentration there was evidence of DNA fragmentation indicating apoptosis induction in Vero cells with a 50% increase in DNA fragmentation compared to control at the highest STX concentration tested (3 nM). The results demonstrated no significant changes in the frequency of micronucleated binucleated cells in N2A and Vero cells exposed to STX, indicating the absence of genotoxicity under these test conditions. There was no apparent cellular necrosis as evidenced by a lack of formation of multinucleated cells. In conclusion, data reported herein demonstrate that STX produced death of both cell types tested through an apoptotic process.


Evaluation of dietary metabolizable energy concentrations on meat quality and lipid metabolism-related gene expression in yellow-feathered chickens.
This study evaluated the effects of different dietary metabolizable energy (ME) concentrations on the meat quality, carcass traits, volatile flavour and lipid metabolism-related gene expression levels in yellow-feathered chickens. In total, 600 Huxu female chickens aged 90 days were randomly assigned to six dietary treatments, each with 10 replicates of 10 birds. During the finisher phase, the birds were fed diets containing 2880 (low), 2940, 3000, 3060, 3120 and 3180 (high) kcal ME/kg. The results showed that the average daily gain of chickens increased as the dietary ME concentration increased, while the feed to gain improved (p < 0.05), and the intramuscular fat content of breast muscle increased (p < 0.05). The energy concentration had no effect on the breast muscle pH (45 min and 24 h), colour parameter (L*) or percentage of drip loss (p > 0.05), but the shear force values decreased significantly (p < 0.05). The diameter and area of the breast muscle fiber decreased and the muscle fibre density increased as the dietary ME concentration increased (p < 0.05). The highest ME concentration (3180 kcal) increased the percentages of aldehydes (hexanal, heptanal, 2,4-nonadienal, octanal, nonanal and 2-decenal), alcohols (2-nonen-1-ol, trans-2-undecen-1-ol, 7-hexadecenal, 2-hexyl-1-decanoal and n-nonadecanol-1,3,7,11-trimethyl-1-dodecanol), alkanes (2,6-dimethyl-heptadecane) and carboxylic acids (9-hexadecenoic acid), but reduced the percentages of octadecanal, octadecane, heneicosane and tetradecanal (p < 0.05). In addition, the mRNA gene expression levels of fatty acid-binding protein 3 and apolipoprotein B were significantly upregulated in the liver, whereas that of cholesteryl ester transfer protein was significantly downregulated. In conclusion, increasing the ME diet to 3180 kcal/kg significantly improved the quality and flavour of the meat from yellow-feathered broilers. our finding may help poultry producers to improve the taste of meat by regulating genes related to lipid metabolism, thereby achieving the flavour and taste characteristics preferred by consumers.


Anemia in adults: a contemporary approach to diagnosis.
There are numerous ways of classifying the causes of anemia, and no one way is necessarily superior to another. It is equally important to appreciate the differences in the approaches to diagnosis between children and adults, men and women, and persons of different ethnic backgrounds. Regardless of the specific algorithm followed in evaluating anemia, it is essential that easily remediable causes such as nutritional deficiencies, hemolysis, and anemia of renal insufficiency are identified early and treated appropriately. In general, the differential diagnosis of anemia can be substantially narrowed by subcategorization into "microcytic," "normocytic," and "macrocytic" subtypes on the basis of mean corpuscular volume. However, such classification is a starting point and not infallible. Each category then can be deciphered using a stepwise approach that utilizes readily accessible laboratory tests.


Analysis and in vivo disruption of the gene coding for calmodulin in Schizosaccharomyces pombe.
Calmodulin is a low molecular weight calcium-binding protein that modulates many enzyme systems in eukaryotes. We have cloned the gene encoding calmodulin from the fission yeast, Schizosaccharomyces pombe, by using synthetic oligonucleotide probes that correspond to three distinct regions of Tetrahymena calmodulin. A 1.6-kilobase (kb) DNA fragment that hybridized to all of them contains a gene whose deduced product possesses 74% amino acid homology with bovine calmodulin. This gene, which is unique in the S. pombe genome and is named cam1, encodes 149 amino acids excluding the first methionine and is transcribed into mRNA of 1.2-kb length. It has an intron that apparently starts immediately after the initiation codon and is 126 bp long. S. pombe calmodulin exhibits more homology to vertebrate calmodulin than to that of the budding yeast, Saccharomyces cerevisiae. Gene disruption experiments revealed that cam1 gene function is essential for vegetative growth of S. pombe. Spores bearing disrupted cam1 halt growth soon after germination and rarely carry out the first cell division, indicating that calmodulin does not exist in excess in those cells.


Minor Flavanones from Erythrina abyssinica
Four new prenylated flavanones, abyssinone-V 4'-methyl ether (1) and abyssinoflavanones IV (2), V (3), and VI (4), have been isolated as minor flavanones from the African medicinal plant, Erythrina abyssinica, together with a known flavanone, sigmoidin D. The structure elucidation of compounds 1-4 by spectroscopic studies is described.


Low-melting ionic solids: versatile materials for ion-sensing devices.
Ionic liquids that melt slightly above room temperature (and may be called low-melting ionic solids, LMISs) were used as sensing materials for the detection of ions in aqueous solutions. A simple procedure based on the consecutive melting and further solidification of the LMISs was applied to prepare solid-contact ion-sensitive electrodes. A potentiometric response toward a number of anions was observed, and the possibility of altering the selectivity by incorporating additional ionophores into the LMIS matrix was demonstrated.


Hypothyroidism reduces content and increases in vitro release of pro-thyrotropin-releasing hormone peptides from the median eminence.
To determine the effect of thyroid status on proTRH-derived peptide processing and secretion, the content and release of TRH and prepro-TRH25-50 (PYE27), as well as somatostatin (SRIF) from median eminence (ME) or olfactory lobe (OL) tissue was studied in the rat. In hypothyroid animals treated by thyroidectomy (Tx), the ME content of TRH and PYE27 was reduced by more than 50%; further, when compared with euthyroid controls there was a significant 2-fold enhancement of the in vitro release of these peptides from ME fragments in response to depolarizing concentrations (60 mM) of potassium. Hyperthyroidism (T4 treatment) caused either no change or an increase in the ME content of these peptides and their response to K+ in vitro did not differ from control animals. The OL content of TRH and PYE27 was unaffected by thyroid status. SRIF levels in both ME and OL as well as in vitro secretion from the ME did not change with either Tx or T4 treatment. The ratio of TRH/PYE27 secretion throughout release and content studies remained stable at 3:1 to 4:1. These findings support the view that TRH in the hypothalamus but not OL is regulated by thyroid hormone. In this location hypothyroidism enhances not only pro TRH synthesis but also release of TRH and another proTRH-derived peptide. The consistent ratio of TRH/PYE27 suggests that regulation of TRH production by thyroid hormone occurs predominantly at the transcriptional level and not through posttranslation processing.


Influence of surgical preparation on the in-vivo response of osteochondral defects.
Cartilage has an extremely poor capacity to heal, which has lead to intensive research into biomaterials and tissue engineering for the purpose of regenerating cartilage in vivo. Many of these techniques have shown great promise in vitro; however, the results do not always carry across to the in-vivo scenario. Healthy cartilage autografts often do not integrate with the adjacent cartilage, suggesting that cartilage is rarely capable of healing even under ideal conditions. It is hypothesized in this study that the surgical creation of defects in cartilage causes significant damage to the adjacent tissues, leading to further degradation of the cartilage and poor outcome for the repair in general. This study compares the healing response of osteochondral defects created with either a punch or a drill in the weight-bearing region of the sheep knee at 4 and 26 weeks following surgery. The use of a drill to create the defect creates a more aggressive inflammatory response at 4 weeks compared with a punch. However, by 26 weeks, defects created with a punch scored higher on the O'Driscoll cartilage grading scale. Tissue damage at the time of surgery plays an important part in the sequence of events for healing of cartilage defects. This knowledge will help to characterize and refine the ovine model for cartilage regeneration and may have an influence on surgical technique and instrumentation for clinical cartilage repair.


[Diagnostic problems in the case of dementia with coexisting hematological disorders].
The case report presents a 61-year-old female patient with profound dementia and coexisting hematological disorders suggesting Addison-Biermer's disease. Two possible diagnoses were considered; dementia of Alzheimer type or dementia secondary to pernicious anaemia. This case report reflects diagnostic difficulties concerning dementia syndrome in the presence of other causative factors.


MDSCs are involved in the protumorigenic potentials of GM-CSF in colitis-associated cancer.
Chronic inflammation is thought to be a major driving force for the development of colitis-associated colorectal cancer (CAC). As one member of proinflammatory cytokine family, granulocyte macrophage colony-stimulating factor (GM-CSF) has been identified to play a key role in CAC pathogenesis recently. The underlying mechanisms, however, remain largely unknown. In this study, we found that myeloid-derived suppressor cells (MDSCs) accumulated increasingly in the lesions during the progression from colitis to cancer, which was critical for CAC formation. Importantly, this MDSC accumulation was controlled by GM-CSF. MDSC number decreased significantly in GM-CSF-deficient mice suffering from CAC induction, and transfusion of MDSCs from wild-type CAC-bearing mice into GM-CSF-deficient counterparts led to recurrence of CAC. Furthermore, the supernatants of CAC lesions or GM-CSF alone was sufficient to differentiate hematopoietic precursors into MDSCs. Addition of neutralizing anti-GM-CSF antibody impaired the MDSC-differentiating effects of the supernatants of CAC lesions. Overall, these findings shed new insights into the mechanisms of GM-CSF underlying CAC development, by inducing/recruiting CAC-promoting MDSCs. Blocking GM-CSF activity or MDSC function may represent new therapeutic strategies for CAC in clinic.


Altered gastric emptying and prevention of radiation-induced vomiting in dogs.
The relation between radiation-induced vomiting and gastric emptying is unclear and the treatment of this condition is not established. We explored, therefore, (a) the effect of cobalt 60 irradiation on gastric emptying of solids and liquids and (b) the possibility of preventing radiation-induced vomiting with the dopamine antagonist, domperidone. Twenty dogs were studied on two separate days, blindly and in random order, after i.v. injection of either a placebo or 0.06 mg/kg domperidone. On a third day, they received 8 Gy (800 rads) whole body irradiation with cobalt 60 gamma-rays after either placebo (n = 10) or domperidone (n = 10). Before each study, each dog was fed chicken liver tagged in vivo with 99mTc-sulfur colloid (solid marker), and water containing 111In-diethylenetriamine pentaacetic acid (liquid marker). Dogs were placed in a Pavlov stand for the subsequent 3 h and radionuclide imaging was performed at 10-min intervals. Irradiation produced vomiting in 9 of 10 dogs given placebo but only in 1 of 10 dogs pretreated with domperidone (p less than 0.01). Gastric emptying of liquids and solids was significantly suppressed by irradiation (p less than 0.01) after both placebo and domperidone. These results demonstrate that radiation-induced vomiting is accompanied by suppression of gastric emptying. Furthermore, domperidone prevents vomiting produced by ionizing radiation but does not alter the accompanying delay of gastric emptying.


Comparative Characteristics of Napsin A, TTF 1 and EGFR Mutation Expression in Mucinous Lung Cell Carcinomas.
Invasive mucinous lung adenocarcinomas are rare and account for 2%-10% of all lung adenocarcinoma cases. It is believed that Napsin A exhibits a weaker expression in mucinous adenocarcinomas compared with TTF1, but such correlation is still poorly researched. The aim of the study was to determine the frequency of mucinous to nonmucinous adenocarcinomas and compare specificity and sensitivity of monoclonal Napsin A with TTF1 in mucinous adenocarcinomas and define the frequency of EGFR mutations. Eighty-four resected lung carcinomas were prospectively evaluated. All biopsies were analysed with p63, TTF1, monoclonal Napsin A, CK7, CK20 and CDX2 and were studied with real-time PCR technology. In resected material we detected 49/84 (58.3%) adenocarcinomas and selected 21 mucinous adenocarcinomas out of 46 non-mucinous adenocarcinomas (45.6%). The most common pattern of mucinous adenocarcinomas is papillary - 24% and colloidal - 24%, followed by acinar - 19.2% and lepidic - 19.2%. mNapsin A was positive in 18/21 (85.7%) mucinous adenocarcinomas v/s 17/21 TTF1 positive (80.9%). EGFR mutations were detected in 3/21 cases with mucinous adenocarcinomas (14.3%): mucinous papillary, mucinous acinar and "salivary gland-like". Our study demonstrates a high proportion of primary mucinous lung adenocarcinomas to primary non-mucinous adenocarcinomas. Sensitivity and specificity of mNapsin A and TTF1 did not show significant difference in pulmonary mucinous and non-mucinous adenocarcinomas, as mNapsin A gave greater sensitivity to mucinous adenocarcinomas. Our results indicate the same mutation frequency of EGFR in mucinous adenocarcinomas as mutation frequency detected in non-mucinous adenocarcinomas in the Bulgarian region.


Experimental study on dynamics of coherent structures formed by inertial solid particles in three-dimensional periodic flows.
We present experimental results obtained under normal gravity on the dynamics of solid particles in periodic oscillatory thermocapillary-driven flows in a non-isothermal liquid bridge made of decane. Inertial particles of different densities and in the size range approximately 0.75-75 μm are able to form stable coherent structures (particle accumulation structures, or PASs). Two image processing techniques were developed and successfully applied to compute time required for an ensemble of particles to form a structure. It is shown that the formation time grows with the decrease of the Stokes number. The observations indicate the probable irrelevance of the memory term for these experiments. Two types of PAS were observed-single (SL-I) and double-loop (SL-II)-which sometimes co-existed. Only large or very dense particles may form an SL-II type structure. A number of novel features of the system were perceived. In some cases, intermittently stable structures emerged (their dynamics is characterized by alternating time intervals during which a structure exists and is destroyed). Whereas in most experiments we observed a conventional symmetric and centered PAS, there were cases when a long-term stable asymmetric structure appeared. Experiments wherein two different types of PAS-forming particles were used simultaneously revealed the destructive role of collisions between the particles on formation of structures.


Helping people to live well with chronic kidney disease.
Reduced glomerular filtration rate and presence of albuminuria are both associated with increased risk of several poor outcomes. People with chronic kidney disease also commonly suffer from lower quality of life than their age-matched peers. The experiences reported by patients with chronic kidney disease include being shocked by the diagnosis, being uncertain about the cause and worrying about progression and future treatment. Issues such as depression, pain and fatigue are common in people with chronic kidney disease. Helping people to live well with a long-term condition like chronic kidney disease should include efforts to reduce the risk of adverse events occurring in the future, and consider what can be done to enhance quality of life now. Clinicians can help by being aware of the patient perspective, communicating clearly and recommending interventions that reduce future risk as well as recognising and treating symptoms. Assessing overall treatment burden is an important component of management and non-pharmacological interventions that may improve mobility, strength and pain should be considered.


Tricho-dento-osseous syndrome and precocious eruption.
Tricho-dento-osseous syndrome (TDO), an uncommon form of ectodermal dysplasia is an autosomal dominant genetic disorder which is characterized by inherited defects in tissues arising from epithelial-mesenchymal interaction. Genetic studies have revealed that it is caused by mutation in the DLX3 gene. TDO presents with a great phenotypic heterogeneity and studies have suggested that this heterogeneity is the result of environmental factors or other genetic modifiers. In this article, we report a case of TDO in which the child had typical clinical features of hair, teeth and bone defects, as seen in TDO. Parents of the child were unaffected. Genetic analysis of the child revealed mutation in DLX3 gene. The child also showed precocious eruption of the permanent molars, a clinical feature which has been rarely reported. We suggest that the precocious eruption seen in TDO is probably due to a markedly increased osteoblastic activity. Key words:Tricho-dento-osseous syndrome, DLX3 gene, precocious eruption.


Depression Treatment among Adults with Multiple Sclerosis and Depression in Ambulatory Care Settings in the United States.
Background. There is little information regarding depression treatment patterns among adults with MS and depression in ambulatory settings at national level in the United States (US). Objectives. The objectives of this study were to identify patterns and predictors of depression treatment in ambulatory settings in US among adults with MS and depression. Methods. A cross-sectional study was conducted by pooling multiple years (2005-2011) of National Ambulatory Medical Care Survey and the outpatient department of the National Hospital Ambulatory Medical Care Survey data. The final study sample was comprised of ambulatory visits among adults with MS and depression. Dependent variable of this study was pharmacological treatment for depression with or without psychotherapy. Predictors of depression treatment were determined by conducting multivariable logistic regression. Results. Out of all ambulatory visits involving MS diagnosis, 20.59% also involved a depression diagnosis. Depression treatment was observed in 57.25% of the study population. Fluoxetine was the most prescribed individual antidepressant. Age and total number of chronic diseases were significant predictors of depression treatment. Conclusion. Approximately six out of ten ambulatory visits involving MS and depression recorded some form of depression treatment. Future longitudinal studies should examine health outcomes associated with depression treatment in this population.


Small-bowel perforation secondary to metastatic carcinoma of the breast.
Perforation from a solitary metastatic lesion of the small bowel is rare. We report a case of acute perforation with no evidence of metastatic disease within the abdomen. Resection of the small bowel was performed.


An emotional Stroop functional MRI study of euthymic bipolar disorder.
To identify the brain regions associated with emotional processing in euthymic bipolar patients. The study examined 12 euthymic bipolar patients using functional magnetic resonance imaging (fMRI) while performing an emotional Stroop (eStroop) task. The task comprised emotionally valent and neutral words presented in alternating blocks that was designed to implicitly induce affect. In conjunction with fMRI, galvanic skin responses (GSR) were measured to monitor arousal. Euthymic bipolar patients had diminished activation in response to the affective stimuli in both cortical and subcortical brain regions when compared with healthy subjects. In particular, patients had less activation in the left ventral prefrontal cortex suggesting a potential trait deficit. Patients were slower to react than healthy controls, but did not differ with respect to accuracy. Euthymic bipolar patients are perhaps constrained in their ability to engage affective processing. Diminished ventral prefrontal cortex activation corroborates previous reports of a potential trait deficit, suggesting that 'all is not well in euthymia', although the effects of medication cannot be overlooked.


Magnetic-actuated "capillary container" for versatile three-dimensional fluid interface manipulation.
Fluid interfaces are omnipresent in nature. Engineering the fluid interface is essential to study interfacial processes for basic research and industrial applications. However, it remains challenging to precisely control the fluid interface because of its fluidity and instability. Here, we proposed a magnetic-actuated "capillary container" to realize three-dimensional (3D) fluid interface creation and programmable dynamic manipulation. By wettability modification, 3D fluid interfaces with predesigned sizes and geometries can be constructed in air, water, and oils. Multiple motion modes were realized by adjusting the container's structure and magnetic field. Besides, we demonstrated its feasibility in various fluids by performing selective fluid collection and chemical reaction manipulations. The container can also be encapsulated with an interfacial gelation reaction. Using this process, diverse free-standing 3D membranes were produced, and the dynamic release of riboflavin (vitamin B2) was studied. This versatile capillary container will provide a promising platform for open microfluidics, interfacial chemistry, and biomedical engineering.


Bioassayable and immunoassayable prolactin responses to thyrotropin-releasing hormone: use of the Nb2 lymphoma cell bioassay.
The peak prolactin response to thyrotropin-releasing hormone (TRH) varies among patients. "Exaggerated" responses have been described and linked to ovulatory dysfunction. Herein we describe our initial observations on bioassayable prolactin (BA-PRL) versus immunoassayable prolactin (RIA-PRL) in women with normal baseline RIA-PRL concentrations but with varying peak RIA-PRL responses to TRH. Twenty-three women of reproductive age with baseline RIA-PRL of =25 ng/mL were administered 500 microg of TRH, and baseline and peak RIA-PRL concentrations were determined. Aliquots of the baseline sample and the sample representing the peak RIA-PRL were used for measuring BA-PRL by means of the Nb2 lymphoma cell bioassay. For each sample, BA/RIA-PRL ratios were determined. Positive correlations were found between peak RIA-PRL and baseline BA/RIA-PRL ratios (P<0.05) and also between baseline and peak BA/RIA-PRL ratios (P<0.001). Negative correlations were found between baseline RIA-PRL and both baseline and peak BA/RIA-PRL ratios (P<0.001 and P<0.05, respectively). We conclude that (1) the lactotroph response to TRH in women with normal RIA-PRL may, in part, be governed by the amount of biologically active prolactin at baseline and (2) the relative proportion of BA-PRL to RIA-PRL produced at baseline is maintained at peak response. Finally, in light of the greater availability of bioactive prolactin in women with exaggerated TRH responses, our findings support the use of bromocriptine in those patients with such responses and ovulatory dysfunction.


Biologically variable bypass reduces enzymuria after deep hypothermic circulatory arrest.
Renal injury is common after open-heart surgery. Cardiopulmonary bypass contributes to the problem. We compared conventional nonpulsatile perfusion (NP) to biologically variable perfusion (BVP), which uses a computer controller to restore physiological beat-to-beat variability to roller pump flow. We hypothesized BVP would decrease renal injury after deep hypothermic circulatory arrest. Pigs were randomly assigned to either BVP (n = 9) or NP (n = 9), cooled, arrested at 18 degrees C (1 hour), reperfused, and rewarmed and maintained normothermic (3 hours). Additional pigs had NP for a similar time as above, but without circulatory arrest (n = 3), or were sham-treated without bypass (n = 3). Hemodynamics, acid-base status, temperature, and urine volumes were measured. Urinary enzyme markers of tubular injury were compared post-hoc for gamma glutamyl transpeptidase, alkaline phosphatase, and glutathione S-transferase and by urine proteomics using mass spectrometry. Urine output at 1 hour after arrest was 250 +/- 129 mL with BVP versus 114 +/- 66 mL with NP (p < 0.02). All three renal enzyme markers were higher with NP after arrest compared with BVP. In animals on bypass without arrest or those sham-treated, no elevations were seen in renal enzymes. Urine proteomics revealed abnormal proteins, persisting longer with NP. Biologically variable perfusion decreased cooling to 21.0 +/- 9.0 minutes versus 31.7 +/- 7.5 minutes (p < 0.002), and decreased rewarming to 22.1 +/- 3.9 minutes versus 31.2 +/- 5.1 minutes (p < 0.002). Biologically variable perfusion improved urine output, decreased enzymuria, and attenuated mass spectrometry urine protein signal with more rapid temperature changes. This strategy could potentially shorten bypass duration and may decrease renal tubular injury with deep hypothermic circulatory arrest.


A novel TRH analog, YM14673, stimulates intracellular signaling systems in the brain more potently than predicted by its pituitary actions.
YM14673, a novel analog of TRH, substituting the (oxo-azetidinyl)carbonyl moiety for the pyroglutamyl moiety of TRH, was synthesized and its brain action was evaluated using rat brain slices in vitro. The addition of YM14673 engendered significant and dose-dependent increases in cyclic AMP formation in the hypothalamus and inositol phosphates (IP) accumulation in the cerebellum. The actions of YM14673 did not statistically differ from those of TRH on brain tissues. However, YM14673 did not show a direct interference with TRH binding in the brain membranes. [3H]YM14673 underwent to the same extent the enzymatic degradation by type II pyroglutamyl aminopeptidase as did TRH. As compared with TRH, YM14673 had a lower activity with respect to stimulation of thyrotropin secretion and accumulation of IP in the anterior pituitaries. The data imply that YM14673 possessed potent stimulatory actions on brain intracellular signaling systems with weak hypophyseal activities. These actions of YM14673 were not due to a direct interference with brain TRH binding or a nature impervious to a TRH-degrading enzyme. The present study indicates that the larger doses of YM14673 than TRH can be used for medical treatment of neurological and neuromuscular disorders without strong influences on pituitary functions.


[Control of arterial hypertension: effectiveness of an intervention performed by family practitioners].
To evaluate the effectiveness of a program to improve hypertension control practices in primary care. Retrospective quasi-experimental study. Three hospital-based family medicine centres (FMCs) Two study groups of 100 randomly-selected adult patients each, who visited the study FMC before implementation of the hypertension program (from April 1, 1983 to March 31, 1984) or afterward (from April 1, 1986 to March 31, 1987). These patients were compared to patients from control FMCs A and B seen during the same time frames (100 patients before and after at FMC A and 60 at FMC B). 1) Educational sessions for physicians to standardize knowledge of the recommendations of the Canadian Hypertension Society on hypertension treatment and 2) specific operational incentives to improve hypertension control, including a reference guide placed in each physician's office, a specific hypertension follow-up form placed with each patient's chart, a recall card file, and hypertension information handouts. Blood pressure measurements recorded in patient charts. Hypertension control is determined from the Canadian Hypertension Society recommendations. The hypertension control rate was 52% in the study group before program implementation and 34.3% afterward (p = 0.01); the corresponding rates in the two control groups moved from 47.4% to 59.8% (p > 0.05) in Group A and from 40.7% to 39.3% (p > 0.05) in Group B. Patients listed in the recall card file were not controlled more frequently (33.3%) than those not listed (35.3%). This intervention did not improve physician practice regarding hypertension control. Clinicians did not follow the protocol as recommended. Physicians must be convinced that a change in their practice is needed before any specific strategies are introduced to support the change. Different suggestions and alternatives related to hypertension management are discussed.


Clinical utility of hyperbaric oxygen therapy in dentistry.
This fuller impact of the use of hyperbaric oxygen therapy within dentistry is taking greater notice with newer research findings. There are new advancements in research regarding postradiotherapy cases, osteonecrosis of the jaw, osteomyelitis, periodontal disease, and dental implants. Hyperbaric oxygen therapy can even be used in conjunction with other procedures such as bone grafting. Although the research and clinical utility has come a long way, there are several complications to be mindful of during the application of hyperbaric oxygen therapy.


Mutational analysis of human BAFF receptor TNFRSF13C (BAFF-R) in patients with common variable immunodeficiency.
BAFF receptor (BAFF-R/BR3/TNFRSF13C) is a recently identified molecule that specifically binds BLyS, a protein belonging to the tumor necrosis factor (TNF) family, and is involved in survival and maturation of B cells. Recent studies have demonstrated that mice defective in BAFF-R gene exhibit an altered profile of the B cell pool, a phenotype observed in BLyS knockout mice as well. These features suggest that mutations in this gene may result in humoral immunodeficiency. To test this hypothesis, we sequenced the BAFF-R gene in 48 patients with common variable immunodeficiency (CVID) along with 57 healthy controls. We have identified three novel variants present at the heterozygous state leading to amino acid substitutions, and have also confirmed the existence of a previously reported intronic variant. The hereby described novel variants were also present in healthy controls and in the healthy patients' parents. These variants do not affect the expression of BAFF-R neither at the mRNA nor at the protein level, suggesting that these variants represent novel polymorphic variants of the BAFF-R gene.


Virtual Screening with Docking Simulations and Biochemical Evaluation of VHY Phosphatase Inhibitors.
Although VH1-related member Y (VHY) phosphatase is responsible for the pathogenesis of neuroinflammatory diseases, no small-molecule inhibitor of VHY has been reported so far. Here we first report eight VHY inhibitors identified from molecular docking-based virtual screening and subsequent enzyme inhibition assays. These inhibitors exhibit good biochemical potencies against VHY, with associated IC50 values ranging from 1 to 9 µM. Because all these inhibitors were also screened in silico for having desirable physicochemical properties as a drug candidate, they deserve further investigation by structure-activity relationship studies to develop new medicines for the treatment of neuroinflammatory diseases. The structural features of VHY-inhibitor interactions relevant to the micromolar-level inhibitory activity are addressed in detail.


Studying Parkinson's disease using Caenorhabditis elegans models in microfluidic devices.
Parkinson's disease (PD) is a progressive neurological disorder associated with the loss of dopaminergic neurons (DNs) in the substantia nigra and the widespread accumulation of α-synuclein (α-syn) protein, leading to motor impairments and eventual cognitive dysfunction. In-vitro cell cultures and in-vivo animal models have provided the opportunity to investigate the PD pathological hallmarks and identify different therapeutic compounds. However, PD pathogenesis and causes are still not well understood, and effective inhibitory drugs for PD are yet to be discovered. Biologically simple but pathologically relevant disease models and advanced screening technologies are needed to reveal the mechanisms underpinning protein aggregation and PD progression. For instance, Caenorhabditis elegans (C. elegans) offers many advantages for fundamental PD neurobehavioral studies including a simple, well-mapped, and accessible neuronal system, genetic homology to humans, body transparency and amenability to genetic manipulation. Several transgenic worm strains that exhibit multiple PD-related phenotypes have been developed to perform neuronal and behavioral assays and drug screening. However, in conventional worm-based assays, the commonly used techniques are equipment-intensive, slow and low in throughput. Over the past two decades, microfluidics technology has contributed significantly to automation and control of C. elegans assays. In this review, we focus on C. elegans PD models and the recent advancements in microfluidic platforms used for manipulation, handling and neurobehavioral screening of these models. Moreover, we highlight the potential of C. elegans to elucidate the in-vivo mechanisms of neuron-to-neuron protein transfer that may underlie spreading Lewy pathology in PD, and its suitability for in-vitro studies. Given the advantages of C. elegans and microfluidics technology, their integration has the potential to facilitate the investigation of disease pathology and discovery of potential chemical leads for PD.


Bilateral orbital varices: an approach to management.
The authors report a case of bilateral orbital varices in a 19-year-old man with a 7-year history of intermittent left proptosis and dystopia. CT demonstrated enhancing lesions with phleboliths and gadolinium-enhanced MRI showed characteristic hyperintense lesions. The asymptomatic right lesion was treated conservatively; the left lesion was excised following intralesional injection of cyanoacrylate. Bilaterality in orbital varices may not be obvious clinically and only diagnosed radiologically. A multidisciplinary approach involving the ophthalmologist and interventional radiologist using intraoperative fluoroscopy enabled accurate characterization, delineation, embolizationm, and excision of the lesion with good hemostatic control.


An evolutionary model of carcinogenesis.
A quantitative model of carcinogenesis based on methods from population biology and game theory demonstrates normal cells in vivo occupy a ridge-shaped maximum in a well-defined tissue fitness landscape, a novel configuration that allows cooperative coexistence of multiple cellular populations. This state, although necessary for development of functioning multicellular organisms, is subject to invasion by fitter, mutant phenotypes permitting somatic evolution of cancer. The model demonstrates carcinogenesis is an emergent phenomenon requiring a sequence of evolutionary steps as cellular proliferation follows successful adaptation to varying environmental constraints. In the initial development of preneoplastic lesions, cellular proliferation is controlled exclusively by interactions with other cells, the extracellular matrix, and soluble or insoluble growth factors so that gain of function mutations in oncogenes, loss of function mutations in tumor suppressor genes, and disruption of normal senescence pathways will permit clonal expansion. This provides explicit selection mechanisms for the mutations depicted in the classical Fearon-Vogelstein model of colorectal carcinogenesis. The model demonstrates neoplastic cellular proliferation can also be promoted by alterations in the somatic landscape that reduce inhibitory signals produced by the normal cells and extracellular matrix. This is consistent with experimental evidence for a strong microenvironmental influence in tumorigenesis independent of genomic changes in the neoplastic populations. However, we find that these changes alone produce only self-limited neoplastic growth because cellular crowding alters system dynamics so that proliferation is dependent on substrate availability. Consequent cellular competition for critical nutrients results in Darwinian selection pressures favoring phenotypes that increase substrate delivery (e.g., angiogenesis) or uptake (e.g., amplify membrane transporters). These previously unknown substrate dynamics in the later stages of carcinogenesis provide a mechanism for evolution of cellular properties typically found in invasive cancers including the angiogenic and glycolytic phenotypes.


Quality of Life in Thyroid Cancer Patients.
Thyroid cancer ranks as the ninth most common cancer worldwide and third in Saudi Arabia. Given thyroid cancer's high incidence, thyroid cancer patients' quality of life (QoL) has been a concern for many years. However, no study has been dedicated to assessing thyroid cancer patients' QoL in Saudi Arabia. Thus, we aimed to fill this gap by assessing thyroid cancer patients' QoL in Saudi Arabia. This cross-sectional study was conducted among thyroid cancer patients attending outpatient clinics at the Diabetes and Endocrinology Centre, King Fahad Specialist Hospital, Qassim region, Saudi Arabia, from 2017 to 2023. The European Organization for the Research and Treatment of Cancer Quality of Life Questionnaire C30 (EORTC QLQ-C30) assessed the patients' QoL through an individual interviewer-administered technique. We used RStudio to perform statistical analysis. Statistical differences between groups based on QoL scores were assessed using the Wilcoxon and Kruskal-Wallis rank-sum tests. Our results showed that for QoL scores of functional domains, the lowest was in social functioning, and the highest was in cognitive functioning. Regarding the subscales of symptoms, the highest scores were fatigue and insomnia. In conclusion, numerous factors affect thyroid cancer patients' QoL. Recognition and appropriate management of these factors will improve the overall QoL; there is a need to understand what is driving these factors in further clinical studies.


Development of Database Assisted Structure Identification (DASI) Methods for Nontargeted Metabolomics.
Metabolite structure identification remains a significant challenge in nontargeted metabolomics research. One commonly used strategy relies on searching biochemical databases using exact mass. However, this approach fails when the database does not contain the unknown metabolite (i.e., for unknown-unknowns). For these cases, constrained structure generation with combinatorial structure generators provides a potential option. Here we evaluated structure generation constraints based on the specification of: (1) substructures required (i.e., seed structures); (2) substructures not allowed; and (3) filters to remove incorrect structures. Our approach (database assisted structure identification, DASI) used predictive models in MolFind to find candidate structures with chemical and physical properties similar to the unknown. These candidates were then used for seed structure generation using eight different structure generation algorithms. One algorithm was able to generate correct seed structures for 21/39 test compounds. Eleven of these seed structures were large enough to constrain the combinatorial structure generator to fewer than 100,000 structures. In 35/39 cases, at least one algorithm was able to generate a correct seed structure. The DASI method has several limitations and will require further experimental validation and optimization. At present, it seems most useful for identifying the structure of unknown-unknowns with molecular weights <200 Da.


Neglect determined by the relative location of responses.
The aim of the present study was to assess the relevance of relative position coding of the effectors in the determination of neglect for the space of response. Two groups of patients were selected: an experimental group, consisting of right brain-damaged patients with left unilateral neglect, and a control group, consisting of right brain-damaged patients without unilateral neglect. These patients participated in two different experiments. In Experiment 1 the stimuli were horizontally aligned to the left and the right of a fixation stimulus; in Experiment 2 they were vertically aligned above and below the fixation stimulus. The stimuli consisted either of the number '1' or the number '2'. In both experiments, the patient was required to give a relative left key press response upon presentation of number '1' and a relative right key press response upon presentation of number '2'. The responses were recorded using the index and middle fingers of the right hand and the response keyboard was located on the right side of body midline. In each experiment there were two hand response conditions: standard and reversed. Under the former condition the hand was in the normal upright position; under the latter, the hand and the response keyboard were rotated by approximately 180 degrees. Under the standard condition, the two effectors were located on the typical anatomical spatial position, i.e. the index finger, which was located on the left relative position, pressed the left key, and the middle finger, which was located on the right relative position, pressed the right key. Under the reversed condition, the positions of the two effectors were reversed, i.e. the middle finger located on the left relative spatial position pressed the left key and the index finger, located on the right relative spatial position, pressed the right key. The results for both experiments showed that neglect patients were always slower to respond when the task required a left key response. The effect was manifest under both the standard and the reversed conditions. The neglect was therefore for the space of response and not for the anatomical spatial position of the effectors.


Dose-response dependencies of OSL dosimeters in conventional linacs and 1.5T MR-linacs: an experimental and Monte Carlo study.
Objective. This work focuses on the optically stimulated luminescence dosimetry (OSLD) dose-response characterization, with emphasis on 1.5T MR-Linacs.Approach. Throughout this study, the nanoDots OSLDs (Landauer, USA) were considered. In groups of three, the mean OSLD response was measured in a conventional linac and an MR-Linac under various irradiation conditions to investigate (i) dose-response linearity with and without the 1.5T magnetic field, (ii) signal fading rate and its dependencies, (iii) beam quality, detector orientation and dose rate dependencies in a conventional linac, (iii) potential MR imaging related effects on OSLD response and (iv) detector orientation dependence in an MR-Linac. Monte Carlo calculations were performed to further quantify angular dependence after rotating the detector around its central axis parallel to the magnetic field, and determine the magnetic field correction factors,kB,Q,for all cardinal detector orientations.Main results. OSLD dose-response supralinearity in an MR-Linac setting was found to agree within uncertainties with the corresponding one in a conventional linac, for the axial detector orientation investigated. Signal fading rate does not depend on irradiation conditions for the range of 3-30 d considered. OSLD angular (orientation) dependence is more pronounced under the presence of a magnetic field. OSLDs irradiated with and without real-time T2w MR imaging enabled during irradiation yielded the same response within uncertainties.kB,Qvalues were determined for all three cardinal orientations. Corrections needed reached up to 6.4%. However, if OSLDs are calibrated in the axial orientation and then irradiated in an MR-Linac placed again in the axial orientation (perpendicular to the magnetic field), then simulations suggest thatkB,Qcan be considered unity within uncertainties, irrespective of the incident beam angle.Significance. This work contributes towards OSLD dose-response characterization and relevant correction factors availability. OSLDs are suitable for QA checks in MR-based beam gating applications andin vivodosimetry in MR-Linacs.


Impact of tall cell variant histology on predicting relapse and changing the management of papillary thyroid carcinoma patients.
There has been much discussion recently about the risk category of tall cell variant (TVC) histology and its effects on the management of papillary thyroid carcinoma (PTC). We, therefore, undertook a retrospective study to compare stage-matched risk factors and recurrence rates between classical PTC (cPTC) patients and patients with TCV histology. A total of 3128 well-differentiated thyroid carcinoma patients who were treated and followed-up for more than 5 years in our clinic from 1995 to 2016 were included in this study. There were 2783 PTC (89%) patients, 1113 (40%) of them were cPTC and 56 (2%) of them were TCV patients. In all stages, the stage-matched incidence of extrathyroidal extension (ETE), lymphovascular invasion and initial lymph node metastases were significantly higher in TCV patients than in cPTC patients (P<0.001). Recurrence was in 10 of 27 patients (37%) with TCV and in 91 of 890 (10%) patients with cPTC diagnosed in stage I (odds ratio (OR)=5.16); in 4 of 6 patients with TCV and 18 of 84 (21%) patients with cPTC in stage II (OR=7.33); in 5 of 6 patients with TCV and 11 of 46 (23%) patients with cPTC in stage III (OR=15.90); and in 13 of 17 patients with TCV and 31 of 93 (33%) patients with cPTC in stage IV (OR=6.50). Stage-matched recurrence rates were found significantly higher in all stages of TCV patients than in cPTC patients (OR=8.49, P<0.001). Recurrence with distant metastases was seen more frequently in TCV patients than in cPTC patients (P<0.001) and treatment of metastatic disease was more difficult in TCV patients. Tall cell variant was an independent poor prognostic factor in papillary thyroid carcinoma patients even if they were diagnosed at early stages of the disease. Patients with tall cell variant histology required more aggressive therapeutic approach and closer follow-up than classical patients.


Pregnancy during therapy for childhood acute lymphoblastic leukemia: two case reports and a review of the literature.
The incidence and consequences of pregnancy during therapy for childhood acute lymphoblastic leukemia (ALL) are largely unknown. To explore the issues involved in this complication of ALL treatment, two recent cases are presented. Two 15-year-old girls with "high risk" ALL became pregnant while receiving maintenance therapy. In one case, the patient experienced a spontaneous abortion at approximately 5 to 6 weeks gestation. The patient completed maintenance therapy and is in remission 8 months after the end of treatment. The second patient, known to be non-compliant during therapy, was found to be 5 months pregnant at the end of maintenance therapy. She developed HELLP syndrome (hemolysis, elevated liver enzymes, and low platelet count), was induced at approximately 34 weeks, and delivered an apparently normal baby girl. Both the patient and her baby continue to do well 10 months after delivery. A variety of factors may influence the incidence of pregnancy during ALL therapy. Gonadal function, which is likely to return to normal during maintenance therapy, may also be affected by alterations in the dose intensity of treatment. Social factors may also alter the incidence of pregnancy. Adverse effects on the fetus are more likely to occur in the first trimester, depending on the drug or drugs used. Although all chemotherapies may have mutagenic and teratogenic effects, they do not invariably cause abnormalities. Survival of adolescents who become pregnant during treatment does not appear to be adversely affected when therapy is not modified or discontinued.


Delta-9-tetrahydrocannabinol decreases masticatory muscle sensitization in female rats through peripheral cannabinoid receptor activation.
This study investigated whether intramuscular injection of delta-9-tetrahydrocannabinol (THC), by acting on peripheral cannabinoid (CB) receptors, could decrease nerve growth factor (NGF)-induced sensitization in female rat masseter muscle; a model which mimics the symptoms of myofascial temporomandibular disorders. Immunohistochemistry was used to explore the peripheral expression of cannabinoid receptors in the masseter muscle while behavioural and electrophysiology experiments were employed to assess the functional effects of intramuscular injection of THC. It was found that CB1 and CB2 receptors are expressed by trigeminal ganglion neurons that innervate the masseter muscle and also on their peripheral endings. Their expression was greater in TRPV1-positive ganglion neurons. Three days after intramuscular injection of NGF, ganglion neuron expression of CB1 and CB2, but not TPRV1, was decreased. In behavioural experiments, intramuscular injection (10 μL) of THC (1 mg/mL) attenuated NGF-induced mechanical sensitization. No change in mechanical threshold was observed in the contralateral masseter muscles and no impairment of motor function was found after intramuscular injections of THC. In anaesthetized rats, the same concentration of THC increased the mechanical thresholds of masseter muscle mechanoreceptors. Co-administration of the CB1 antagonist AM251 blocked the effect of THC on masseter muscle mechanoreceptors while the CB2 antagonist AM630 had no effect. These results suggest that reduced inhibitory input from the peripheral cannabinoid system may contribute to NGF-induced local myofascial sensitization of mechanoreceptors. Peripheral application of THC may counter this effect by activating the CB1 receptors on masseter muscle mechanoreceptors to provide analgesic relief without central side effects. Our results suggest THC could reduce masticatory muscle pain through activating peripheral CB1 receptors. Peripheral application of cannabinoids could be a novel approach to provide analgesic relief without central side effects.


Multidimensional human capital formation in a developing country: Health, cognition and locus of control in the Philippines.
Economic success depends on multiple human capital stocks whose production is interrelated and occurs over many life stages. Yet, much empirical work fails to account for human capital's multidimensional nature and limits its focus to specific childhood stages. Using longitudinal data from the Philippines, I estimate a model of multidimensional human capital formation from birth through adulthood where health, cognitive, and noncognitive dimensions are jointly produced. I examine during which developmental stages parental investment is most influential and address the endogeneity of investment using a policy function where investment depends on child characteristics, exogenous conditions at birth and local prices. Findings imply that not only will early human capital disparities persist into adulthood without early remediation but also that cognitive gains yielded from early remediation will be lost without complementary investment in adolescence. Findings further suggest that interventions will be undervalued if their multidimensional effects are not accounted for.


Achieving Predictable Hydraulic Sinus Condensing Using an Innovative Functional Sinus Implant: A Case Report.
Inadequate alveolar bone height/volume and sinus anatomy can complicate the predictable placement of conventional implants and subsequent restoration in posterior maxillary sites. A variety of sinus lift and vertical bone height augmentation materials and techniques have been utilized for this purpose; however, generally they are clinically challenging, demand well-trained surgical proficiency, and require the use of multiple armamentaria. Simultaneously, clinicians must meticulously handle the sinus membrane to achieve elevation without perforation, attain successful bone grafting, and ensure predictable implant osseointegration. The hydraulic sinus condensing (HSC) technique via the osteotomy site is designed to be less invasive than conventional sinus lift techniques, reduce trauma to patients, improve implant osseointegration into grafted alveolar bone, prevent sinus membrane perforation, and enhance the overall efficiency of the associated surgical process. This article briefly reviews the evolution of sinus lift and posterior maxillary implant insertion procedures and presents a case in which the HSC technique and a uniquely designed sinus implant were used.


Liver protection by silymarin: in vitro effect on dissociated rat hepatocytes.
The liver-protective effects of silymarin have been investigated in isolated hepatocytes suspended in vitro in sucrose solutions of graded hypotonicity. Cells preincubated with the drug at the concentration of 10(-4) M were found to be entirely resistant or relatively little affected when exposed to hypotonic solutions which have a lethal effect on a considerable proportion of the control liver cells, as shown by the determination in the cell preparations (1) of the proportion trypan blue-positive (broken) cells, (2) of the protein synthesizing capacity. The protection was reduced but not abolished upon removal of the drug and cell washing. A smaller effect was observed with a drug concentration of 10(-5) M. At 10(-6) M the effect was no longer detectable. Chlorpromazine, a drug which belongs to the group of the so called "unspecific membrane stabilizers", was without effect in this test.


Azilsartan treatment improves insulin sensitivity in obese spontaneously hypertensive Koletsky rats.
Hypertension often coexists with insulin resistance. However, most metabolic effects of the antihypertensive agents have been investigated in nomotensive animals, in which different conclusions may arise. We investigated the metabolic effects of the new angiotensin II type 1 receptor blocker azilsartan using the obese Koletsky rats superimposed on the background of the spontaneously hypertensive rats. Male Koletsky rats were treated with azilsartan (2 mg/kg/day) over 3 weeks. Blood pressure was measured by tail-cuff. Blood biochemical and hormonal parameters were determined by enzymatic or ELISA methods. Gene expression was assessed by RT-PCR. In Koletsky rats, azilsartan treatment lowered blood pressure, basal plasma insulin concentration and the homeostasis model assessment of insulin resistance index, and inhibited over-increase of plasma glucose and insulin concentrations during oral glucose tolerance test. These effects were accompanied by decreases in both food intake and body weight (BW) increase. Although two treatments showed the same effect on BW gain, insulin sensitivity was higher after azilsartan treatment than pair-feeding. Azilsartan neither affected plasma concentrations of triglyceride and free fatty acids, nor increased adipose mRNA levels of peroxisome proliferator-activated receptor (PPAR)γ and its target genes such as adiponectin, aP2. In addition, azilsartan downregulated 11β-hydroxysteroid dehydrogenase type 1 expression. These results show the insulin-sensitizing effect of azilsartan in obese Koletsky rats. This effect is independent of decreases in food intake and BW increase or of the activation of adipose PPARγ. Our findings indicate the possible usefulness of azilsartan in the treatment of metabolic syndrome.


Wake-promoting effects of vagus nerve stimulation after traumatic brain injury: upregulation of orexin-A and orexin receptor type 1 expression in the prefrontal cortex.
Orexins, produced in the lateral hypothalamus, are important neuropeptides that participate in the sleep/wake cycle, and their expression coincides with the projection area of the vagus nerve in the brain. Vagus nerve stimulation has been shown to decrease the amounts of daytime sleep and rapid eye movement in epilepsy patients with traumatic brain injury. In the present study, we investigated whether vagus nerve stimulation promotes wakefulness and affects orexin expression. A rat model of traumatic brain injury was established using the free fall drop method. In the stimulated group, rats with traumatic brain injury received vagus nerve stimulation (frequency, 30 Hz; current, 1.0 mA; pulse width, 0.5 ms; total stimulation time, 15 minutes). In the antagonist group, rats with traumatic brain injury were intracerebroventricularly injected with the orexin receptor type 1 (OX1R) antagonist SB334867 and received vagus nerve stimulation. Changes in consciousness were observed after stimulation in each group. Enzyme-linked immunosorbent assay, western blot assay and immunohistochemistry were used to assess the levels of orexin-A and OX1R expression in the prefrontal cortex. In the stimulated group, consciousness was substantially improved, orexin-A protein expression gradually increased within 24 hours after injury and OX1R expression reached a peak at 12 hours, compared with rats subjected to traumatic brain injury only. In the antagonist group, the wake-promoting effect of vagus nerve stimulation was diminished, and orexin-A and OX1R expression were decreased, compared with that of the stimulated group. Taken together, our findings suggest that vagus nerve stimulation promotes the recovery of consciousness in comatose rats after traumatic brain injury. The upregulation of orexin-A and OX1R expression in the prefrontal cortex might be involved in the wake-promoting effects of vagus nerve stimulation.


University of Puerto Rico - School of Pharmacy: one hundred years of history and service.
The University of Puerto Rico School of Pharmacy was initially established as the Department of Pharmacy at the Río Piedras Campus on September 22, 1913, being the first healthcare program founded in Puerto Rico. The history of the School of Pharmacy reveals a remarkable evolution over a 100 years period in order to respond to the patients' and societal needs. The programs offered by the school have adapted to the wealth of scientific knowledge, emerging technologies, patients' needs and changes in the healthcare system. This historical note highlights the most important milestones of this centennial institution.


Identification of heterozygotic carriers of 21-hydroxylase deficiency: sensitivity of ACTH stimulation tests.
Congenital adrenal hyperplasia due to 21-hydroxylase deficiency is a common autosomal-recessive disorder. To ascertain carrier status, adrenocorticotropin (ACTH) stimulation tests are often used. To determine the sensitivity of ACTH stimulation to detect heterozygotes and to correlate stimulated 17-hydroxyprogesterone responses with molecular genotype, we compared molecular genetic analysis of the 21-hydroxylase (CYP21) gene with 17-hydroxyprogesterone responses at 30 min in 51 individuals. Molecular genotype analysis and ACTH stimulation tests were performed in healthy volunteers (n = 20) and relatives of patients with congenital adrenal hyperplasia (n = 31). Polymerase chain reaction (PCR) amplification, single-strand conformational polymorphism (SSCP) analysis, allele-specific oligonucleotide hybridization (ASOH) analysis, and restriction fragment length polymorphism (RFLP) analysis were utilized to screen for 14 CYP21 mutations which account for >90% of the mutations associated with 21-hydroxylase deficiency. Molecular genotype analysis classified 28 individuals as heterozygotic carriers and 23 individuals as normal for all mutations tested. As a group, the heterozygotes had significantly greater stimulated 17-hydroxyprogesterone responses at 10 and 30 min (P < 0.0005). However, on an individual basis, 14/28 (50%) genotyped heterozygotic carriers had stimulated 17-hydroxyprogesterone concentrations, 17-hydroxyprogesterone/cortisol ratios, and 17-hydroxyprogesterone incremental elevations indistinguishable from the genotyped normal individuals. Thus, a normal 17-hydroxyprogesterone response to ACTH stimulation testing does not exclude carrier status for 21-hydroxylase deficiency. Molecular genotype analysis is a more reliable method to determine 21-hydroxylase heterozygotes.