Profilin-Mediated Actin Allocation Regulates the Growth of Epithelial Microvilli.
Transporting epithelial cells, like those that line the intestinal tract, are specialized for solute processing and uptake. One defining feature is the brush border, an array of microvilli that serves to amplify apical membrane surface area and increase functional capacity. During differentiation, upon exit from stem-cell-containing crypts, enterocytes build thousands of microvilli, each supported by a parallel bundle of actin filaments several microns in length. Given the high concentration of actin residing in mature brush borders, we sought to determine whether enterocytes were resource (i.e., actin monomer) limited in assembling this domain. To examine this possibility, we inhibited Arp2/3, the ubiquitous branched actin nucleator, to increase G-actin availability during brush border assembly. In native intestinal tissues, Arp2/3 inhibition led to increased microvilli length on the surface of crypt, but not villus, enterocytes. In a cell culture model of brush border assembly, Arp2/3 inhibition accelerated the growth and increased the length of microvilli; it also led to a redistribution of F-actin from cortical lateral networks into the brush border. Effects on brush border growth were rescued by treatment with the G-actin sequestering drug, latrunculin A. G-actin binding protein, profilin-1, colocalized in the terminal web with G-actin, and knockdown of this factor compromised brush border growth in a concentration-dependent manner. Finally, the acceleration in brush border assembly induced by Arp2/3 inhibition was abrogated by profilin-1 knockdown. Thus, brush border assembly is limited by G-actin availability, and profilin-1 directs unallocated actin monomers into microvillar core bundles during enterocyte differentiation.


Study on the water structure and blood compatibility of poly(acryloylmorpholine-r-butyl methacrylate).
We have been studying the blood compatibility of polymeric materials from the viewpoint of their water structure, and have proposed that freezable water interacting with polymer molecules plays an important role in determining that compatibility. As we found that water-soluble poly(acryloylmorpholine) interacted with water, resulting in the formation of 'bound water', we newly prepared water-non-soluble poly(acryloylmorpholine-r-butyl methacrylate) (denoted as ACMO co-polymer) with various composition ratios. In addition, the properties of a co-polymer based on N,N-diethylacrylamide (DEA co-polymer), where DEA has a similar chemical structure to ACMO, except that DEA has no ether oxygen, were compared with that of the ACMO co-polymer. Contact angle and DSC analysis revealed that an increase in the content of an N-substituted acrylamide unit in the co-polymers enhanced the hydrophilicity of the polymer and that the hydrophilicity of the ACMO co-polymer was stronger than that of the DEA co-polymer. As for the water structure, it was found that the ACMO co-polymer had a lot of bound water compared to the DEA co-polymer. The difference in these properties between the ACMO and DEA co-polymers was due to the ether oxygen of the morpholine group. At the same time, in vitro blood compatibility tests showed that the ACMO co-polymer exhibited a much better performance than the DEA co-polymer. The water structure and blood compatibility is discussed in detail.


Synthesis and applications of helical polymers with dynamic and static memories of helicity.
This review mainly highlights our studies on the synthesis of one-handed helical polymers with a static memory of helicity based on the noncovalent helicity induction with a helical-sense bias and subsequent memory of the helicity approach that we developed during the past decade. Apart from the previous approaches, an excess one-handed helical conformation, once induced by nonracemic molecules, is immediately retained ("memorized") after the complete removal of the nonracemic molecules, accompanied by a significant amplification of the asymmetry, providing novel switchable chiral materials for chromatographic enantioseparation and asymmetric catalysis as well as a highly sensitive colorimetric and fluorescence chiral sensor. A conceptually new one-handed helix formation in a racemic helical polymer composed of racemic repeating units through the deracemization of the pendants is described.


Risk of Axillary Nerve Injury in Standard Anterolateral Approach of Shoulder: Cadaveric Study.
Introduction: The anterolateral acromion approach of the shoulder is popular for minimally invasive plate osteosynthesis (MIPO) technique. However, there are literatures describing the specific risks of injury of the axillary nerve using this approach. Nevertheless, most of the studies were done with Caucasian cadavers. So, the purpose of this study was to evaluate the risk of iatrogenic axillary nerve injury from using the anterolateral shoulder approach and further investigate the location of the axillary nerve, associated with its location and arm length in the Asian population that have shorter arm length compared to the Caucasian population. Materials and Methods: Seventy-nine shoulders in fourty-two embalmed cadavers were evaluated. The bony landmarks were drawn, and a vertical straight incision was made 5cm from tip of the acromion (anterolateral approach), to the bone. The iatrogenic nerve injury status and the distance between the anterolateral edge of the acromion to the axillary nerve was measured and recorded. Results: In ten of the seventy-nine shoulders, the axillary nerve were iatrogenically injured. The average anterior distance was 6.4cm and the average arm length was 30.2cm. The anterior distance and arm length ratio was 0.2. Conclusion: Our results demonstrated that the recommended safe zone at 5cm from tip of acromion was not suitable with Asian population due to shorter arm length, compared to Caucasian population. The location of axillary nerve could be predicted by 20% of the total arm-length.


Upregulation of Vesicular Glutamate Transporter 2 and STAT3 Activation in the Spinal Cord of Mice Receiving 3,3'-Iminodipropionitrile.
Chronic administration of 3,3'-iminodipropionitrile (IDPN) causes axonal impairment. Although controversy still remains, it has been suggested that IDPN intoxication mimics the axonopathy of amyotrophic lateral sclerosis (ALS). Interestingly, recent studies including our own showed that signal transducer and activator of transcription 3 (STAT3) in spinal α-motoneurons was activated in both IDPN-treated mice and SOD1 G93A mice, a genetic model of familial ALS. Because activation of STAT3 occurs in response to various stimuli, such as axonal injury, ischemia, and excessive glutamate, here we focused on a potential link between phosphorylated STAT3 (pSTAT3, an active form) and vesicular glutamate transporter 2 (VGluT2, a regulator of glutamate storage and release) in IDPN-treated mice and SOD1 G93A mice. Impairment of axonal transport was confirmed by western blot analysis: the expression levels of phosphorylated neurofilament H were elevated in both models. As shown in SOD1 G93A mice, the expression frequencies of VGluT2 in synaptophysin-positive (SYP)+ presynaptic terminals around spinal α-motoneurons were significantly higher in IDPN-treated mice than in vehicle controls. The coverages of spinal α-motoneurons by VGluT2+ presynaptic terminals were more elevated around pSTAT3+ cells than around pSTAT3- cells in IDPN-treated mice and SOD1 G93A mice. Considering that excessive glutamate is shown to be involved in axonal impairment and STAT3 activation, the present results suggest that IDPN-induced upregulation of VGluT2 may result in an increase in glutamate, which might cause axonopathy and induction of pSTAT3. The link between upregulation of VGluT2 and activation of STAT3 via glutamate may represent a common pathological feature of IDPN-treated mice and SOD1 G93A mice.


Differential distribution of a glucuronyltransferase, involved in glucuronoxylan synthesis, within the Golgi apparatus of pea (Pisum sativum var. Alaska).
The subcellular location of a glucuronyltransferase (GT) involved in glucuronoxylan synthesis in pea (Pisum sativum) has been investigated. Most of the GT activity was found in the Golgi fraction, but activity was also detected in the plasma-membrane fraction. Separation of Golgi membranes on a shallow continuous sucrose density gradient resulted in three distinct subfractions, with GT activity being confined to Golgi membranes of a density similar to that of smooth endoplasmic reticulum. The differential distribution of GT within the Golgi stack indicates that glucuronoxylan synthesis occurs in specific cisternae and that there is functional compartmentalization of the Golgi with respect to hemicellulose biosynthesis.


The efficacy of antidepressants in the treatment of late-life depression.
This review addresses the question of whether there is evidence that antidepressants are more efficacious than placebo in the treatment of late-life depression and what is the rate of response that physician and patient can expect when antidepressant medication is prescribed in a typical clinical setting. To date, 5 placebo-controlled and 10 comparison trials have study designs of sufficient rigor to provide evidence of antidepressant efficacy and effectiveness in the treatment of late-life depression. The results suggest that antidepressant medications are more effective than placebo. However, placebo-controlled trials are not a simple comparison of only medication versus placebo. Rather, the amount of nonspecific psychosocial interventions included in these trials is considerable and often not systematically measured. Trial design also affects outcome: response and remission rates in comparison trials consistently are 20% to 30% higher than those reported in placebo-controlled trials. Clinical trials do not consistently assess the many moderators that are believed to affect treatment outcome in late-life depression, and therefore, comparisons across studies are problematic because of an inability to determine whether patient samples are truly comparable. For future clinical trials to have maximal relevance, study design should evolve to reflect as closely as possible a typical clinical setting especially with respect to frequency and duration of patient visits.


18α-Glycyrrhetinic acid (GA) ameliorates fructose-induced nephropathy in mice by suppressing oxidative stress, dyslipidemia and inflammation.
Excessive fructose (FRU) intake can result in insulin resistance and metabolic disorder, which are related to renal injury.18α-Glycyrrhetinic acid (GA) is a bioactive component mainly extracted from Glycyrrhiza radix, and has anti-oxidant and anti-inflammatory activities. However, its effects on FRU-induced renal injury still remain unclear. In this study, we found that 18α-GA treatments could significantly ameliorate the cell viability in FRU-treated tubule epithelial cells, accompanied with improved mitochondrial membrane potential. Furthermore, reactive oxygen species (ROS) accumulation in FRU-stimulated cells was markedly reduced by 18α-GA, which were associated with the activation of nuclear factor (erythroid-derived-2)-like 2 (Nrf-2) and the blockage of MAPKs signaling. Additionally, dyslipidemia detected in FRU-treated cells was greatly inhibited by 18α-GA. We also found that 18α-GA significantly ameliorated FRU-induced inflammation in cells through reducing the expression of pro-inflammatory cytokines and chemokine. The anti-inflammatory effects regulated by 18α-GA were mainly related to the repression of nuclear factor-κB(NF-κB) signaling. Furthermore, the protective effects of 18α-GA against ROS production, lipid accumulation and inflammation were verified in renal tissues from FRU-challenged mice, consequently improving metabolic disorder and kidney injury. Taken together, these findings demonstrated that 18α-GA exerted renal protective effects through reducing oxidative stress, lipid deposition and inflammatory response, and thus could be considered as a promising therapeutic strategy for metabolic stress-induced kidney injury.


[Diagnosis of shunt dysfunction after transjugular intrahepatic portosystemic shunt by color Doppler ultrasonography: usefulness change of maximum flow velocity in portal vein].
To clarify whether color doppler ultra sonography (CDUS) is useful for the diagnosis of shunt dysfunction after transjugular intrahepatic portosystemic shunt (TIPS), we investigated CDUS in 20 patients who were diagnosed as shunt dysfunction and 12 patients with patent shunt by follow-up portal pressure measurements. In patients with shunt dysfunction, maximum flow velocity in the portal vein (PV-Vmax) and maximum flow velocity in the shunt were significantly decreased in comparison to just after TIPS placement, but not in patients with patent shunt. Grade of color image of the shunt flow and flow direction in the portal vein branches were not significantly different between the shunt dysfunction group and the patent shunt group. Comparing the just after TIPS to the follow-up phase, the decreasing rate of PV-Vmax was negatively correlated with the increasing rate of portal vein pressure caused by shunt dysfunction. CDUS could detect shunt dysfunction in 95% sensitivity, 83% specificity and 91% overall accuracy on the basis of a diagnostic criterion for shunt stenosis : more than 40% decrease of PV-Vmax from just after TIPS insertion. These results suggest that follow-up observation using CDUS is useful for the diagnosis of shunt dysfunction after TIPS procedure, and PV-Vmax is an especially important indicator to detect shunt dysfunction.


Combination versus alternating chemotherapy in small-cell lung cancer.
The major obstacles in the treatment of small-cell lung cancer (SCLC) are tumor recurrence and the emergence of drug resistance to chemotherapeutic agents. Using alternating non-cross-resistant combination regimens is an attractive strategy, but the efficacy and toxicities are still controversial. Previously untreated SCLC patients were randomly assigned to standard CEOV therapy (cyclophosphamide, epirubicin, vincristine = oncovin, and etoposide = VP-16) or CEOV alternating with CAM therapy (cyclophosphamide, ACNU and methotrexate). Chemotherapy was repeated every 4 weeks for a total of eight cycles. Complete and partial responders were administered thoracic irradiation. Between July 1987 and July 1993, 150 patients were enrolled and 123 were evaluable. There was no significant superiority in the treatment outcome for CEOV-CAM therapy over CEOV therapy in terms of overall response rate (51% vs 71%), complete response rate (9% vs 35%), median survival time (9.5 months vs 12.0 months) and 2-year survival rate (7% vs 13%). Both patients with limited disease and extensive disease had equal response rates to the CEOV regimen (70% and 71%, respectively). However, patients with limited disease had a significantly higher response rate to the CEOV-CAM regimen than did patients with extensive disease (65% vs 36%, p = 0.04). Toxicities for both treatment arms were comparable. Patients who achieved complete or partial response received thoracic irradiation and had significantly longer survival than responders who did not receive thoracic irradiation (14.0 months vs 12.0 months, p = 0.02). There were equal chances of tumor recurrence in the chest and at distant sites (66% vs 67%) in the CEOV arm. However, in the CEOV-CAM arm, tumors were more likely to occur in the chest than in distant sites (86% vs 36%) (p = 0.0016). We concluded that alternating chemotherapy provides no therapeutic advantage compared with four-regimen combination chemotherapy in terms of tumor response, median survival and 2-year survival rate for SCLC patients. Early thoracic irradiation should be applied to achieve better local control and less distant recurrence for chemotherapy responders.


Increasing late diagnosis in HIV infection in South Korea: 2000-2007.
The number of Koreans diagnosed with human immunodeficiency virus (HIV) infections is increasing annually; however, CD4+ T-cell counts at diagnosis have decreased. The purpose of the present study was to identify clinical and epidemiologic associations with low CD4+ T-cell counts at the time of HIV diagnosis in a Korean population. Data from 2,299 HIV-infected individuals with initial CD4+ T-cell counts measured within 6 months of HIV diagnosis and reason for HIV testing were recorded and measured from 2000 to 2007. Data were selected from the database of the Korea Centers for Disease Control and Prevention. Late diagnosis was defined by CD4+ T-cell counts <200 cells/mm3. Reasons for HIV testing were analyzed using logistic regression including epidemiologic variables. A total of 858 individuals (37.3%) were included in the late diagnosis group. Individuals with a late diagnosis were older, exposed through heterosexual contact, and demonstrated clinical manifestations of acquired immunodeficiency syndrome (AIDS). The primary reason for HIV testing was a routine health check-up (41%) followed by clinical manifestations (31%) of AIDS. The proportion of individuals with a late diagnosis was higher in individuals tested due to clinical symptoms in public health centers (adjusted odds ratio [AOR], 17.3; 95% CI, 1.7-175) and hospitals (AOR, 4.9; 95% CI, 3.4-7.2) compared to general health check-up. Late diagnosis annually increased in individuals diagnosed by voluntary testing both in public health centers (PHCs, P = 0.017) and in hospitals (P = 0.063). Routine testing due to risky behaviors resulted in earlier detection than testing secondary to health check-ups, although this difference was not statistically significant (AOR, 0.7; P = 0.187). Individuals identified as part of hospital health check-ups more frequently had a late diagnosis (P = 0.001) HIV infection was primarily detected by voluntary testing with identification in PHCs and by testing due to clinical symptoms in hospitals. However, early detection was not influenced by either voluntary testing or general health check-up. It is important to encourage voluntary testing for early detection to decrease the prevalence of HIV infection and AIDS progression.


Recurrent laryngeal never monitoring versus non-monitoring in parathyroid surgery.
Although intraoperative neural monitoring (IONM) is well established in thyroid surgery, it is less commonly analyzed in parathyroid operations. This study presents the results of IONM for primary and secondary hyperparathyroidism surgery. We retrospectively assessed 270 patients with primary hyperparathyroidism (PHPT), 53 patients with secondary hyperparathyroidism (SHPT), and 300 patients with thyroid cancer from June 2010 to June 2022 in one hospital in China. The follow-up was 12 months. Demographic, electromyography data from IONM, laboratory, and clinical information were collected. Laryngoscopy was collected from 109 patients with PHPT in whom IONM was not used. All groups were assessed by Pearson's chi-square test and Fisher's exact probability method to verify the relationship between parathyroid size and location, duration of surgery, preoperative concordant localization, laryngeal pain, IONM outcomes, cure rate, and RLN injury. Visual analog scale (VAS) assessed laryngeal pain. RLN outcomes were measured according to nerves at risk (NAR). The study comprehended 918 NAR, that is 272, 105, 109, and 432 NAR for PHPT, SHPT with IONM, PHPT without IONM, and thyroid surgery control group, respectively. IONM successfully prevented RLN injury (P<0.001, P=0.012): Fifteen (5.51%) RLNs experienced altered nerve EMG profiles during surgery, and five (1.84%) experienced transient RLN injury in PHPT patients. Five (4.76%) RLNs were found to have altered EMG profiles during surgery, and one (0.95%) RLN had a transient RLN injury in SHPT patients. There was no permanent nerve injury (0.00%) in this series. There was no association between location, gland size, preoperative concordant localization, cure rate, duration of surgery, and IONM (P >0.05). Duration of surgery was associated with postoperative pharyngeal discomfort (P=0.026, P=0.024). Transient RLN injury was significantly lower in patients with PHPT who underwent IONM than in those who did not. Intraoperative neuromonitoring played an effective role in protecting the recurrent laryngeal nerve (P=0.035). Compared with parathyroidectomy, thyroidectomy had a higher rate of RLN injury (5.32%, P<0.001). IONM for SHPT and PHPT offers rapid anatomical gland identification and RLN functional results for effective RLN protection and reduced RLN damage rates.


CORECLUST: identification of the conserved CRM grammar together with prediction of gene regulation.
Identification of transcriptional regulatory regions and tracing their internal organization are important for understanding the eukaryotic cell machinery. Cis-regulatory modules (CRMs) of higher eukaryotes are believed to possess a regulatory 'grammar', or preferred arrangement of binding sites, that is crucial for proper regulation and thus tends to be evolutionarily conserved. Here, we present a method CORECLUST (COnservative REgulatory CLUster STructure) that predicts CRMs based on a set of positional weight matrices. Given regulatory regions of orthologous and/or co-regulated genes, CORECLUST constructs a CRM model by revealing the conserved rules that describe the relative location of binding sites. The constructed model may be consequently used for the genome-wide prediction of similar CRMs, and thus detection of co-regulated genes, and for the investigation of the regulatory grammar of the system. Compared with related methods, CORECLUST shows better performance at identification of CRMs conferring muscle-specific gene expression in vertebrates and early-developmental CRMs in Drosophila.


Evaluation of 3D Printed Soft Robots in Radiation Environments and Comparison With Molded Counterparts.
Robots have an important role during inspection, clean-up, and sample collection in unstructured radiation environments inaccessible to humans. The advantages of soft robots, such as body morphing, high compliance, and energy absorption during impact, make them suitable for operating under extreme conditions. Despite their promise, the usefulness of soft robots under a radiation environment has yet to be assessed. In this work, we evaluate the effectiveness of soft robots fabricated from polydimethylsiloxane (PDMS), a common fabrication material, under radiation for the first time. We investigated gamma-induced mechanical damage in the PDMS materials' mechanical properties, including elongation, tensile strength, and stiffness. We selected three radiation environments from the nuclear industry to represent a wide range of radiation and then submerged a 3D printed hexapus robot into a radiation environment to estimate its operation time. Finally, to test the reliability of the 3D printed soft robots, we compared their performances with molded counterparts. To analyze performance results in detail, we also investigated dimensional errors and the effects of fabrication methods, nozzle size, and print direction on the stiffness of PDMS material. Results of this study show that with increasing exposure to gamma irradiation, the mechanical properties of PDMS decrease in functionality but are minimally impacted up to 20 kGy gamma radiation. Considering the fractional changes to the PDMS mechanical properties, it is safe to assume that soft robots could operate for 12 h in two of the three proposed radiation environments. We also verified that the 3D printed soft robots can perform better than or equal to their molded counterparts while being more reliable.


Inverse Problem of Ultrasound Beamforming With Sparsity Constraints and Regularization.
Ultrasound (US) beamforming is the process of reconstructing an image from acquired echo traces on several transducer elements. Typical beamforming approaches, such as delay-and-sum, perform simple projection operations, while techniques using statistical information also exist, e.g., adaptive, phase coherence, delay-multiply-and-sum, and sparse coding approaches. Inspired by the feasibility and success of inverse problem (IP) formulations in several image reconstruction problems, such as computed tomography, we herein devise an IP approach for US beamforming. We define a linear forward model for the synthesis of the beamformed image, and solve its IP thanks to several intuitive and physics-based constraints and regularization terms proposed. These reflect the prior knowledge about the spectra of carrier signal and spatial coherence of modulated signal. These constraints admit effective formulation through sparse representations. Our proposed method was evaluated for plane-wave imaging (PWI) that transmits unfocused waves, enabling high frame rates with large field of view at the expense of much lower image quality with conventional beamforming techniques. Results are evaluated in numerical simulations, as well as tissue-mimicking phantoms and in vivo data provided by PWI challenge in medical US. The best results achieved by our proposed techniques are 0.39-mm full-width at half-maximum for spatial resolution and 16.3-dB contrast-to-noise ratio, using a single plane-wave transmit.


Osteoblast Response of Additively Manufactured Zirconia and Alumina-Toughened Zirconia.
Zirconia ceramics have been widely used in dentistry. Herein, we assess the surface morphology, surface texture, and osteoblast response of additively manufactured zirconia and alumina-toughened zirconia (ATZ) in comparison with titanium. The surface roughness, contact angle, and surface microstructure of titanium sandblasted with large-grit alumina and subsequently acid-etched using 18% HCl and 49% H2SO4 (SLA-titanium), uniaxially pressed zirconia (UP zirconia), additively manufactured zirconia (AM zirconia), and additively manufactured ATZ (AM ATZ) were investigated. Moreover, the cell viability, alkaline phosphatase (ALP) activity, and gene expression of type I collagen on these materials were evaluated. The data were statistically analyzed using one-way ANOVA with Tukey's post hoc test. SLA-titanium showed the highest surface roughness and contact angle. The other three materials showed comparable surface roughness and contact angles. Micro- and nanoroughness were observed on the surface of SLA-titanium. UP zirconia and AM zirconia had similar surface morphologies. The cell viability, ALP activity, and gene expression of type I collagen on AM zirconia were comparable to or better than those on SLA-titanium. Our results indicate that AM zirconia is a promising material for zirconia dental implants.


Effects of month of kidding, parity number, and litter size on milk yield of commercial dairy goats in Australia.
The aim of this observational study was to identify the influence of key nongenetic factors such as month of kidding, parity, and litter size on milk yield and composition of Australian dairy goats throughout lactation. The study was conducted over 4 consecutive kidding seasons from June 2016 to March 2017. Data from 940 lactations of Saanen goats from a commercial herd were used to observe the effects of month of kidding, parity number, and litter size on total milk yield (L/goat) in early lactation (kidding to 90 d in milk; DIM), mid lactation (91-180 DIM), and late lactation (181-270 DIM), cumulative milk yield (from kidding to 270 DIM; CMY), average lactation length, proportion (%) of does reaching their target lactation length (270 DIM), somatic cell count (SCC), and percentages of milk fat and protein in early lactation. The mean herd responses throughout the entire study were as follows: CMY = 519 L/goat; lactation length = 233 d, with 70% of does reaching 270 DIM; milk fat = 4.2%; milk protein = 2.9%; and SCC = 6.2 × 105 cells/mL. Average milk production peaked in February and was lowest in June (2.4 vs. 1.8 L/goat per day, respectively). Milk yield was affected by month of kidding, parity number, and litter size in all phases of lactation. November kidders had the greatest CMY, and March kidders had the lowest CMY. March kidders had the shortest lactation length and the lowest proportion of does reaching 270 DIM. June kidders had the longest lactation length, whereas September kidders had the highest proportion of does reaching 270 DIM. Maximum milk yield was attained in third parity. Goats in fourth or greater parity had the shortest lactation length, the lowest proportion of does reaching 270 DIM, and the highest SCC. Goats delivering single kids had lower CMY, lower SCC, and higher percentages of fat and protein than does delivering multiple kids. Our findings indicate that milk yield was primarily influenced by month of kidding, and the effects of month of kidding on milk yield were accentuated during mid lactation. However, the effects of month of kidding on milk yield varied significantly among parities.


Phosphotransferase-mediated regulation of carbohydrate utilization in Escherichia coli K12: the nature of the iex (crr) and gsr (tgs) mutations.
Mutants of Escherichia coli K12 defective in the gene iex (crr) no longer utilize glucose or N-acetylglucosamine in preference to lactose, but competition between either of these sugars and another that also enters by a phosphotransferase (PT) mechanism is not affected. In this they differ from gsr (tgs) mutants. In gsr mutants, glucose does not exclude any other sugar, though N-acetylglucosamine still does so. In gsr mutants that are also ptsM the phosphoenolpyruvate-dependent phosphorylation of glucose or methyl alpha-glucoside is reduced by 90%: N-acetylglucosamine phosphorylation is not affected. The iex mutation does not affect the phosphorylation of either of these compounds. The wild-type alleles iex+ and gsr+ are dominant in lambda heterozygotes. Glucose inhibits the lactose permease of wild-type cells, but only when the permease is present in low amounts. The inhibition is also relieved (1) by induction of another transport system that is subject to regulation by the iex system or (2) by an iex mutation. We suggest that the iex gene specifies a protein that, in cells transporting certain sugars by a PT mechanism, acts to inhibit active transport systems. The protein is present in limiting concentration in the cell, sufficient only to inhibit the basal, uninduced, level of the active transport systems. In consequence the inducer (or its precursor) may be excluded from the cell and induction thus prevented.


The Sensitivity of Clinical Outcomes to Centration on the Light-Constricted Pupil for a Shape-Changing Corneal Inlay.
To assess the clinically acceptable range of inlay decentration with respect to the light-constricted pupil center and the coaxially sighted corneal light reflex (CSCLR) for an inlay (Raindrop Near Vision Inlay; ReVision Optics, Inc., Lake Forest, CA) that reshapes the anterior corneal surface. In this retrospective, observational cohort study of 115 patients with emmetropic or low hyperopic presbyopia who were implanted with a shape-changing corneal inlay, visual acuity, task performance (in good and dim light), reports of halos and glare, and satisfaction data were collected from the preoperative and 3-month postoperative examinations. Inlay centration with respect to the pupil center and CSCLR was determined from the center of the inlay effect derived from iTrace (Tracey Technologies, Houston, TX) wavefront measurements. Multivariate regression models assessed the influence of inlay position on visual outcomes. On average, monocular uncorrected near visual acuity (UNVA) improved 4.9 ± 1.7 lines in the treated eye, with no loss in binocular distance vision. Eighty-three percent of implants were centered radially within 0.5 mm of the pupil center. Multivariate analysis of decentration with respect to both the pupil center and CSCLR revealed no significant interaction with the above clinical outcomes, with the exception of UNVA in the treated eye (all P > .05, α = 0.05). For decentration of less than 0.75 mm, the change in UNVA was less than 1 line. Distance and near visual acuity, task performance, severity of halos and glare, and satisfaction were independent of radial decentration of the Raindrop Near Vision Inlay of less than 0.75 mm from the light-constricted pupil. [J Refract Surg. 2018;34(3):164-170.].


The effects of continuous and pulsed exposures of suspended clay on the survival, growth, and reproduction of Daphnia magna.
Suspended sediments are a natural component of aquatic ecosystems, but anthropogenic activity such as land development can result in significant increases, especially after rain events. Continuous exposures of suspended clay and silt have been shown to affect growth and reproduction of Cladocera, leading to a decrease in population growth rate. The mechanism of clay toxicity in these filter-feeding organisms is clogging of the gut tract, resulting in decreased food uptake and assimilation. When placed in clean water, daphnids can purge clay from their gut and recover. In many surface waters, aquatic organisms experience episodic exposures of high concentrations of suspended solids driven by rain events. However, little is known about the consequences of pulsed exposures on individuals and populations. The objective of the present study was to characterize the effects of continuous and pulsed exposures of natural and defined clays on survival, growth, and reproduction of Daphnia magna. Two defined clays, montmorillonite and kaolinite, as well as clay isolated from the Piedmont region of South Carolina, USA, were used. Continuous exposures of clays elicited a dose dependent decrease in survival. Toxicity varied depending on clay source with montmorillonite > natural clay > kaolinite. Pulsed exposures caused a decrease in survival in a 24 h exposure of 734 mg/L kaolinite. Exposure to 73.9 mg/L also caused an increase in the time to gravidity, although there was not a corresponding decrease in neonate production over 21 d. No significant effects resulted from 12 h exposures even at 730 mg/L, almost 10 times the 24-h reproductive effects concentration. This suggests that exposure duration impacted toxicity more than exposure concentration in these pulsed exposures.


Low-dose exposure of glyphosate-based herbicides disrupt the urine metabolome and its interaction with gut microbiota.
Glyphosate-based herbicides (GBHs) can disrupt the host microbiota and influence human health. In this study, we explored the potential effects of GBHs on urinary metabolites and their interactions with gut microbiome using a rodent model. Glyphosate and Roundup (equal molar for glyphosate) were administered at the USA glyphosate ADI guideline (1.75 mg/kg bw/day) to the dams and their pups. The urine metabolites were profiled using non-targeted liquid chromatography-high resolution mass spectrometry (LC-HRMS). Our results found that overall urine metabolite profiles significantly differed between dams and pups and between female and male pups. Specifically, we identified a significant increase of homocysteine, a known risk factor of cardiovascular disease in both Roundup and glyphosate exposed pups, but in males only. Correlation network analysis between gut microbiome and urine metabolome pointed to Prevotella to be negatively correlated with the level of homocysteine. Our study provides initial evidence that exposures to commonly used GBH, at a currently acceptable human exposure dose, is capable of modifying urine metabolites in both rat adults and pups. The link between Prevotella-homocysteine suggests the potential role of GBHs in modifying the susceptibility of homocysteine, which is a metabolite that has been dysregulated in related diseases like cardiovascular disease or inflammation, through commensal microbiome.


Block-dependent sedation during epidural anaesthesia is associated with delayed brainstem conduction.
Neuraxial anaesthesia produces a sedative and anaesthetic-sparing effect. Recent evidence suggests that spinal cord anaesthesia modifies reticulo-thalamo-cortical arousal by decreasing afferent sensory transmission. We hypothesized that epidural anaesthesia produces sensory deafferentation-dependent sedation that is associated with impairment of brainstem transmission. We used brainstem auditory evoked potentials (BAEP) to evaluate reticular function in 11 volunteers. Epidural anaesthesia was induced with 2-chloroprocaine 2%. Haemodynamic and respiratory responses, sensory block level, sedation depth and BAEP were assessed throughout induction and resolution of epidural anaesthesia. Sedation was evaluated using verbal rating score (VRS), observer's assessment alertness/sedation (OAA/S) score, and bispectral index score (BIS). Prediction probability (PK) was used to associate sensory block with sedation, as well as BIS with other sedation measures. Spearman's rank order correlation was used to associate block level and sedation with the absolute and interpeak BAEP latencies. Sensory block level significantly predicted VRS (PK=0.747), OAA/S score (PK=0.748) and BIS. BIS predicted VRS and OAA/S score (PK=0.728). The latency of wave III of BAEP significantly correlated with sedation level (rho=0.335, P<0.01) and sensory block (rho=0.394, P<0.01). The other BAEP parameters did not change during epidural anaesthesia. Haemodynamic and respiratory responses remained stable throughout the study. Sedation during epidural anaesthesia depends on sensory block level and is associated with detectable block-dependent alterations in the brainstem auditory evoked responses. Sensory deafferentation may reduce CNS alertness through mechanisms related to brainstem neural activity.


Cerebral blood flow determinations using fluorescent microspheres: variations on the sedimentation method validated.
We validate a modification of the sedimentation method for measuring fluorescent microspheres (FM) that improves the determination of regional cerebral blood flow (rCBF). Our FM method for rCBF determination is compared to the radioactive microspheres (RM) method for rCBF measurement by simultaneous injection of one radioactive and two fluorescent labeled doses, at two separate time points, into the left ventricle of a pig. The pig was killed, the brain and spinal cord removed, and divided into 92 pieces averaging 0.83 g. Our modifications to FM analysis by sedimentation includes: 2 instead of 1 week of autolysis, pellet washing with 1% Triton X-100 instead of 0.25% Tween 80, phosphate buffer addition during rinse, fluorescent dye extraction using 2-ethoxyethylacetate instead of 2-(2-ethoxyethoxy)ethyl acetate and polypropylene instead of glass tubes. Comparing rCBF using Sc46 RM, to yellow-green and orange FM, yielded mean differences of 0.026 and 0.021 ml/min per piece, respectively. Sn(113) RM compared to blue-green and scarlet FM gave mean differences of -0.010 and 0.137 ml/min per piece, respectively. All RM-FM differences, except those for scarlet FM, are within acceptable limits. This assay provides a reliable method for determining rCBF.


Acetyl coenzyme A carboxylase: filamentous nature of the animal enzymes.
Acetyl coenzyme A carboxylases purified from several animal tissues exist as enzymatically active polymeric filaments of high molecular weight and have simillar electron microscopic, hydrodynamic, and catalytic properties. These filaments reversibly dissociate into inactive protomers of uniform size. Their re-assembly into catalytically active filaments is promoted by the presence of an allosteric activator.


Adaptability and stability of conilon coffee in areas of high altitude.
In view of the predicted models of global climate change and differences in prices and production costs, there is increased interest in Coffea canephora cultivation in areas of high altitude. However, this species is sensitive to low temperatures, where genotypes vary regarding adaptation/tolerance mechanisms, demonstrating genotype x environment interaction. The aim of this study was to evaluate the stability and adaptability of C. canephora varieties in high-altitude areas. The experiments were carried out in February 2004, in Bom Jesus do Itabapoana, Rio de Janeiro State, Brazil, at an altitude of 725 m. Four clonal varieties (EMCAPA 8111, EMCAPA 8121, EMCAPA 8131, and EMCAPA 8151) and five harvests (2006 to 2010) were used. The experimental design was randomized blocks with four treatments and six plots, with 12 plants in each plot and spaced 2.5 x 1.2 m. Adaptability and stability parameters were determined using methods based on nonparametric analysis and analysis of variance. The results showed that the EMCAPA 8131 had the best performance according to stability and adaptability parameters and may be promising for high-altitude regions.


Women With Synchronous or Metachronous Lung and Ovarian Cancer: A Multi-Institutional Report.
Double diagnosis of lung cancer (LC) and ovarian cancer (OC) is rare. Here, we describe patients with synchronous/metachronous LC and OC to identify common clinical and pathological patterns. Clinical, pathological and molecular data of patients diagnosed and treated at 30 European Institutions from 2008 to 2018 were retrieved and analysed. Whenever tissue was available, centralized pathology revision was performed. A total of 19 cases were found; one was excluded at pathology revision. Most LCs were adenocarcinomas (15/18) and most OCs were high-grade serous (15/18) carcinomas. Of the 9 patients analysed, 7 carried oncogene-addicted LC (4 EGFR, 1 B-RAF and 2 ALK) and five out of 7 carried BRCA mutations. One patient with a germline-BRCA1 mutation received olaparib, resulting in a durable response of both malignancies. Median overall survival was 33 months. In our series, most synchronous/metachronous LCs and OCs showed genetic alterations. Further analyses with wide NGS panel could shed light on the biological mechanisms driving their occurrence.


Peroxidasin and eosinophil peroxidase, but not myeloperoxidase, contribute to renal fibrosis in the murine unilateral ureteral obstruction model.
Renal fibrosis is the pathological hallmark of chronic kidney disease (CKD) and manifests as glomerulosclerosis and tubulointerstitial fibrosis. Reactive oxygen species contribute significantly to renal inflammation and fibrosis, but most research has focused on superoxide and hydrogen peroxide (H2O2). The animal heme peroxidases myeloperoxidase (MPO), eosinophil peroxidase (EPX), and peroxidasin (PXDN) uniquely metabolize H2O2 into highly reactive and destructive hypohalous acids, such as hypobromous and hypochlorous acid. However, the role of these peroxidases and their downstream hypohalous acids in the pathogenesis of renal fibrosis is unclear. Our study defines the contribution of MPO, EPX, and PXDN to renal inflammation and tubulointerstitial fibrosis in the murine unilateral ureteral obstruction (UUO) model. Using a nonspecific inhibitor of animal heme peroxidases and peroxidase-specific knockout mice, we find that loss of EPX or PXDN, but not MPO, reduces renal fibrosis. Furthermore, we demonstrate that eosinophils, the source of EPX, accumulate in the renal interstitium after UUO. These findings point to EPX and PXDN as potential therapeutic targets for renal fibrosis and CKD and suggest that eosinophils modulate the response to renal injury.


Visceral leishmaniasis on the Indian Subcontinent: Efficacy of fipronil-based cattle treatment in controlling sand fly populations is dependent on specific aspects of sand fly ecology.
Visceral leishmaniasis (VL) is a deadly disease transmitted by the sand fly Phlebotomus argentipes on the Indian subcontinent, with a promising means of vector control being orally treating cattle with fipronil-based drugs. While prior research investigating the dynamic relationship between timing of fipronil-based control schemes and the seasonality of sand flies provides insights into potential of treatment on a large scale, ecological uncertainties remain. We investigated how uncertainties associated with sand fly ecology might affect our ability to assess efficacy of fipronil-based control schemes. To do this, we used a previously-described, individual-based, stochastic sand fly model to quantify how uncertainties associated with 1) the percentage of female sand flies taking blood meals from cattle, and 2) the percentage of female sand flies ovipositing in organic matter containing feces from treated cattle might impact the efficacy of fipronil-based sand fly control schemes. Assuming no prior knowledge of sand fly blood meal and oviposition sites, the probabilities of achieving effective sand fly population reduction with treatments performed 3, 6 and 12 times per year were ≈5-22%, ≈27-36%, and ≈46-54%, respectively. Assuming ≥50% of sand flies feed on cattle, probabilities of achieving efficacious control increased to ≈8-31%, ≈15-42%, and ≈52-65%. Assuming also that ≥50% of sand flies oviposit in cattle feces, the above probabilities increased further to ≈14-53%, ≈31-81%, and ≈89-97%. Our assessments of the efficacy of fipronil-based cattle treatments in controlling sand fly populations depend on our assumptions regarding key aspects of sand fly ecology. Assessments are most sensitive to assumptions concerning the percentage of sand flies ovipositing in feces of treated cattle, thus emphasizing the importance of identifying sand fly oviposition sites. Our results place the evaluation of fipronil-based cattle treatment within a broader ecological context, which could aid in the planning and execution of a largescale field trial.


Acetate-mediated growth inhibition in sterol 14alpha-demethylation-deficient cells of Candida albicans.
Candida albicans is a fungus thought to be viable in the presence of a deficiency in sterol 14alpha-demethylation. We showed in a strain of this species that the deficiency, caused either by a mutation or by an azole antifungal agent, made the cells susceptible to growth inhibition by acetate included in the culture medium. Studies with a mutant demonstrated that the inhibition was complete at a sodium acetate concentration of 0.24 M (20 g/liter) and was evident even at a pH of 8, the latter result indicating the involvement of acetate ions rather than the undissociated form of acetic acid. In fluconazole-treated cells, sterol profiles determined by thin-layer chromatography revealed that the minimum sterol 14alpha-demethylation-inhibitory concentrations (MDICs) of the drug, thought to be the most important parameter for clinical purposes, were practically identical in the media with and without 0.24 M acetate and were equivalent to the MIC in the acetate-supplemented medium. The acetate-mediated growth inhibition of azole-treated cells was confirmed with two additional strains of C. albicans and four different agents, suggesting the possibility of generalization. From these results, it was surmised that the acetate-containing medium may find use in azole susceptibility testing, for which there is currently no method capable of measuring MDICs directly for those fungi whose viability is not lost as a result of sterol 14alpha-demethylation deficiency. Additionally, the acetate-supplemented agar medium was found to be useful in detecting reversions from sterol 14alpha-demethylation deficiency to proficiency.


Two-photon correlations in a mixture of gaussian and laser light.
Measurement of the two-photon time distribution in a mixture of light from a He-Ne laser and a Gaussian source of the same central frequency agrees well with the prediction of coherence theory when the laser is represented by a single coherent state. The peak in the distribution near zero time difference is made up of contributions due to the second-order interference between pairs of Gaussian photons and between laser and Gaussian photons.


The "angina-producing" myocardial segment: An approach to the interpretation of results of coronary bypass surgery.
The first 153 cases of saphenous vein aortocoronary bypass surgery performed at The Johns Hopkins Hospital were reviewed. Eighty-eight percent of the 140 late survivors reported significant symptomatic improvement. Seventy-one unselected patients consented to complete reevaluation at a mean interval of 6.1 months postoperatively. Vein bypass patency in this group was 66 percent. Eighty-two percent of these 71 patients had improved performance on electrocardiographic stress testing. There were no significant differences between hemodynamic status on pre- and postoperative studies regardless of the status of vein bypass patency. Repeat coronary angiography revealed a 30 percent incidence of new total occlusions of the intrinsic coronary circulation. Segmental wall motion on ventriculography was improved in 12 percent, unchanged in 50 percent and decreased in 38 percent of the segments analyzed. Localized electrocardiographic changes compatible with myocardial damage developed in the immediate postoperative period in 45 of these patients (63 percent); in 38 of the 45, these changes corresponded to new angiographic abnormalities. Physiologic mechanisms underlying symptomatic improvement were sought by identification of an "angina-producing" myocardial segment. In most instances, alleviation of angina could be related to either: (1) a patent bypass graft into an unchanged intrinsic coronary artery with presumed increased blood flow to the distal coronary segment, or (2) occlusion of the bypass graft and the corresponding coronary artery with probable infarction of previously ischemic myocardium. Current criteria for the selection of patients undergoing coronary bypass surgery are reviewed in the light of these findings. It is concluded that relief of disabling angina is the major indication for surgery. Prevention of myocardial infarction and improvement in left ventricular function are at present less reliable objectives.


Neonatal characteristics and perinatal complications in neonates with Down syndrome.
The annual rate of Down syndrome (DS) births in Jerusalem is stable, regardless of prenatal screening, and diagnostic measures. We aimed to evaluate our historical cohort for obstetrical characteristics and the neonatal course and complications. We reviewed computerized medical files of neonates with the diagnosis of DS born in the four main hospitals in Jerusalem between the years 2000 and 2010 and evaluated for maternal history and primary neonatal hospitalization. A total of 403 neonates were diagnosed with DS. The average maternal age was 35.6 years, 73% were born via spontaneous vaginal delivery. In all gestational ages, the mean birth weight and head circumference percentiles were significantly lower than the general population (P < 0.001 for both) and at each week the HC percentile was lower than the weight percentile (P < 0.0001), worse among males. Mortality during the primary hospitalization was 3.7%. The most common anomalies were cardiac (79%) with either congenital defects or functional abnormalities, neither influenced the length of hospitalization. The main reasons for prolonged hospitalization were prematurity and anomalies of other (non-cardiac) organs. Common perinatal complications included respiratory failure or need for oxygen supplementation (32%), hyperbilirubinemia (23%), sepsis (6.4%), and feeding difficulties (13%). About 84% were fed by human milk; of those, two thirds were exclusively breast-fed and one third were supplemented with infant formula. In conclusion, infants with DS were small for gestational age with relatively reduced head circumference. Despite the increased rate of congenital anomalies and perinatal complications, most infants were discharged home in good medical condition and were exclusively breastfed.


Investigating Instructor Talk among Graduate Teaching Assistants in Undergraduate Biology Laboratory Classrooms.
Instructor Talk-noncontent and nonlogistical language that is focused on shaping the classroom learning environment-is a recently defined variable that may play an important role in how undergraduates experience courses. Previous research characterized Instructor Talk used by faculty teaching in biology lecture classrooms. However, graduate teaching assistants (GTAs) and laboratory classrooms represent critical factors in undergraduate education, and Instructor Talk in this context has yet to be explored. Here, we present findings analyzing Instructor Talk used by GTAs teaching in undergraduate biology laboratory classrooms. We characterized the Instructor Talk used by 22 GTA instructors across 24 undergraduate biology laboratory courses in the context of a single, urban, Hispanic-serving and Asian American and Pacific Islander-serving Institution. We found that Instructor Talk was present in every course studied, GTAs with pedagogical training and prior teaching experience used more Instructor Talk than those without, and GTAs teaching laboratory courses used more Instructor Talk than previous observations of faculty teaching lecture courses. Given the widespread use of Instructor Talk and its varying use across contexts, we predict that Instructor Talk may be a critical variable in teaching, specifically in promoting equity and inclusion, which merits continued study in undergraduate science education.


(2R,6R)-Hydroxynorketamine Alleviates Electroconvulsive Shock-Induced Learning Impairment by Inhibiting Autophagy.
Learning impairment after electroconvulsive therapy (ECT) is common. Ketamine, an anesthetic used for ECT, has been demonstrated to attenuate cognitive impairment after ECT. However, the mechanism by which ketamine occurs in this case is still unknown. We aimed to explore the role of ketamine metabolite (2R,6R)-hydroxynorketamine [(2R,6R)-HNK] in the protection against learning impairment and investigate whether autophagy is involved in the protective effect. A rat depression model received electroconvulsive shock (ECS; simulated ECT in animal models) daily for 3 days. The Morris water maze was used to assess the spatial learning function of the rats. Western blotting was used to detect the expression of Beclin-1, light chain (LC)3-II/LC3-I, p62, mammalian target of rapamycin (mTOR), and p-mTOR in the hippocampus. The escape latency for the maze in the ECS group was significantly longer than that in the sham ECS group (P=0.042). Meanwhile, the escape latency in the (2R,6R)-HNK+ECS group was significantly shorter than that in the ECS group (P=0.005). The LC3-II/LC3-I ratio and Beclin-1 expression level significantly increased, and the p62 expression level significantly decreased in the ECS group, compared with those in the sham ECS group (all P<0.001). The (2R,6R)-HNK+ECS group showed lower LC3-II/LC3-I ratio (P<0.001) and Beclin-1 expression level (P<0.001) and higher p62 (P<0.001) and p-mTOR expression levels (P=0.048) than did the ECS group. After small-molecule enhancer of rapamycin 28 (SMER28) administration, the role of (2R,6R)-HNK in protecting against learning impairment and inhibiting autophagy was abrogated, showing no difference in the escape latency; the difference in the LC3-II/LC3-I ratio and p62 expression level between the SMER28+(2R,6R)-HNK+ECS and ECS groups was not as significant as that between the (2R,6R)-HNK+ECS and ECS groups (P<0.05-0.01 vs P<0.001). (2R,6R)-HNK yields cognitive protection by suppressing autophagy through the mTOR signaling pathway in the ECS-treated rat hippocampus.


The effect of geometrical parameters of skin brachytherapy patch source on depth dose distribution using Monte Carlo simulation.
Brachytherapy of superficial skin tumors using beta-emitting sources is a method that has been investigated by some researchers in both simulation and experimental studies with promising results. In the current study, the effect of geometrical parameters of some relevant radionuclides including Y-90, Re-188, P-32, and Ho-166 on the depth dose distribution in skin tissue has been investigated through Monte Carlo simulation. MCNPX Monte Carlo code was employed to model the above-mentioned patch sources in cylindrical format and then the effect of patch geometrical parameters including the source-to-skin distance (SSD), patch thickness, and patch diameter on depth dose distribution was assessed through modeling and calculation of the dose inside a cubic phantom mimicking the skin tissue. The obtained results demonstrated that increasing the SSD, patch thickness, and patch diameter (with the same activity) will reduce the depth dose distribution. Changing the SSD has a more significant effect on the dose gradient within the depth than other geometrical parameters. It was also observed that the effect of patch diameter on the skin-delivered dose gets less sensible as the patch size goes beyond the range of beta radiation inside tissue. Finally, it can be concluded that the patch source geometrical parameters can affect the depth dose distribution inside the skin tissue. This fact may be of concern regarding the delivery of a high radiation dose in a single treatment session. Therefore, variations of patch source geometrical parameters should be considered during the skin dose calculation plan.


A novel evaluation of residue and protein volumes by means of Laguerre tessellation.
Amino acids control the protein folding process and maintain its functional fold. This study underlines the interest of the Laguerre tessellation to determine relevant amino acid volumes in proteins. Previous studies used a limited number of proteins and only buried residues. The present computations improve the method and results on three main points: (i) a large, high-quality updated and refined data bank of proteins is used; (ii) all residues are taken into account, including those at the protein surface, thanks to (iii) the addition of a realistic solvent. The new values of the average and standard deviation of amino acid volumes show significant corrections with respect to previous studies. Another issue of the method is the polyhedral protein/water interface area (PIA) which quantifies the exposure of atoms or residues to the solvent. We propose this PIA as a new, parameter-free, alternative for measuring accessibility. The comparison with NACCESS is satisfactory; however, the methods disagree in pointing out buried residues: where NACCESS evaluates to zero, the exposure given by PIA ranges from 0 to 20%. Variations of average residue volumes have been analyzed under several conditions, e.g., how they depend on protein size and on secondary structure environments. As it is based on strong mathematical grounds and on numerous high-quality protein structures, our work gives a reliable methodology and up-to-date values of amino acid volumes and surface accessibility.


Synergistic metabolism in hybrid corn indicates bottlenecks in the carotenoid pathway and leads to the accumulation of extraordinary levels of the nutritionally important carotenoid zeaxanthin.
Lutein and zeaxanthin cannot be synthesized de novo in humans, and although lutein is abundant in fruit and vegetables, good dietary sources of zeaxanthin are scarce. Certain corn varieties provide adequate amounts because the ratio of endosperm β:ε lycopene cyclase activity favours the β-carotene/zeaxanthin branch of the carotenoid pathway. We previously described a transgenic corn line expressing the early enzymes in the pathway (including lycopene β-cyclase) and therefore accumulating extraordinary levels of β-carotene. Here, we demonstrate that introgressing the transgenic mini-pathway into wild-type yellow endosperm varieties gives rise to hybrids in which the β:ε ratio is altered additively. Where the β:ε ratio in the genetic background is high, introgression of the mini-pathway allows zeaxanthin production at an unprecedented 56 μg/g dry weight. This result shows that metabolic synergy between endogenous and heterologous pathways can be used to enhance the levels of nutritionally important metabolites.


Rationale and design of the Medication adherence Improvement Support App For Engagement-Blood Pressure (MedISAFE-BP) trial.
Hypertension is a major contributor to the health and economic burden imposed by stroke, heart disease, and renal insufficiency. Antihypertensives can prevent many of the harmful effects of elevated blood pressure, but medication nonadherence is a known barrier to the effectiveness of these treatments. Smartphone-based applications that remind patients to take their medications, provide education, and allow for social interactions between individuals with similar health concerns have been widely advocated as a strategy to improve adherence but have not been subject to rigorous testing. The MedISAFE-BP study is a prospective, randomized control trial designed to evaluate the impact on blood pressure and medication adherence of an mhealth application (Medisafe). Four hundred thirteen patients with uncontrolled hypertension have been enrolled and randomized in a 1:1 fashion to usual care or to the use of the Medisafe mhealth platform. Patients will be followed up for 12 weeks and the trial's co-primary outcomes will be change in systolic blood pressure and self-reported medication adherence. The MedISAFE-BP trial is the first study to rigorously evaluate an mhealth application's effect on blood pressure and medication adherence. The results will inform the potential effectiveness of this simple system in improving cardiovascular disease risk factors and clinical outcomes.


A mammalian type I fatty acid synthase acyl carrier protein domain does not sequester acyl chains.
The synthases that produce fatty acids in mammals (FASs) are arranged as large multidomain polypeptides. The growing fatty acid chain is bound covalently during chain elongation and reduction to the acyl carrier protein (ACP) domain that is then able to access each catalytic site. In this work we report the high-resolution nuclear magnetic resonance (NMR) solution structure of the isolated rat fatty acid synthase apoACP domain. The final ensemble of NMR structures and backbone (15)N relaxation studies show that apoACP adopts a single, well defined fold. On conversion to the holo form, several small chemical shift changes are observed on the ACP for residues surrounding the phosphopantetheine attachment site (as monitored by backbone (1)H-(15)N correlation experiments). However, there are negligible chemical shift changes when the holo form is modified to either the hexanoyl or palmitoyl forms. For further NMR analysis, a (13)C,(15)N-labeled hexanoyl-ACP sample was prepared and full chemical shift assignments completed. Analysis of two-dimensional F(2)-filtered and three-dimensional (13)C-edited nuclear Overhauser effect spectroscopy experiments revealed no detectable NOEs to the acyl chain. These experiments demonstrate that unlike other FAS ACPs studied, this Type I ACP does not sequester a covalently linked acyl moiety, although transient interactions cannot be ruled out. This is an important mechanistic difference between the ACPs from Type I and Type II FASs and may be significant for the modulation and regulation of these important mega-synthases.


[Latero-lateral meso-caval anastomosis in portal hypertension. Report concerning 20 cases (author's transl)].
A new technique of latero-lateral meso-caval technique for the treatment of portal hypertension is described and 20 consecutive patients have been treated using this method with success. There were no operative deaths or severe complications. Follow up over periods ranging from one and a half to three years indicates that all the patients are well. Oesophageal varices disappeared in 4 cases and 8 patients have been able to return to work. The author briefly discusses the technique of porto-systemic shunt and the necessary features of the procedure. The advantages of this technique are summarised. If there is a wide gap between the superior mesenteric vein and the inferior vena cava, the author suggests taking a graft in the area in order to perform a meso-caval bypass by interposition.


Low-temperature sputtered nickel oxide compact thin film as effective electron blocking layer for mesoscopic NiO/CH3NH3PbI3 perovskite heterojunction solar cells.
We introduce the use of low temperature sputtered NiOx thin film, which substitutes the PEDOT-PSS and solution-processed NiOx as an effective electron blocking layer for mesoscopic NiO/CH3NH3PbI3 perovskite solar cells. The influences of film thickness and oxygen doping on the photovoltaic performances are scrutinized. The cell efficiency has been improved from 9.51 to 10.7% for devices using NiOx fabricated under pure argon atmosphere. With adequate doping under 10% oxygen flow ratio, we achieved power conversion efficiency of 11.6%. The procedure is large area scalable and has the advantage for cost-effective perovskite-based photovoltaics.


General practice patients--their readiness to quit smoking.
This article examines the prevalence of smoking among general practice patients and assesses their stage of readiness to quit. Descriptive study involving eight general practice registrars working in teaching practices in metropolitan Sydney (New South Wales) who surveyed 1069 consecutive patients over 16 years of age to determine their smoking status; and for smokers, their stage of readiness to stop smoking. Of these patients 375 (35%) were current smokers, with smoking more common among men (40%) than women (33%). Proportions of smokers in each stage of change were: 137 in precontemplation (36.5%), 158 in contemplation (42%) and 79 in preparation (21%). The majority of patients in preparation (67%) and contemplation (53%) were assessed as willing to further discuss their smoking, whereas only 16% of those in the contemplation stage were willing. Smoking rates among general practice patients were higher than in community samples. Most of the smokers were either contemplating or preparing to quit, and the majority of smokers in these groups were willing to receive advice about smoking cessation.


Correcting population stratification in genetic association studies using a phylogenetic approach.
The rapid development of genotyping technology and extensive cataloguing of single nucleotide polymorphisms (SNPs) across the human genome have made genetic association studies the mainstream for gene mapping of complex human diseases. For many diseases, the most practical approach is the population-based design with unrelated individuals. Although having the advantages of easier sample collection and greater power than family-based designs, unrecognized population stratification in the study samples can lead to both false-positive and false-negative findings and might obscure the true association signals if not appropriately corrected. We report PHYLOSTRAT, a new method that corrects for population stratification by combining phylogeny constructed from SNP genotypes and principal coordinates from multi-dimensional scaling (MDS) analysis. This hybrid approach efficiently captures both discrete and admixed population structures. By extensive simulations, the analysis of a synthetic genome-wide association dataset created using data from the Human Genome Diversity Project, and the analysis of a lactase-height dataset, we show that our method can correct for population stratification more efficiently than several existing population stratification correction methods, including EIGENSTRAT, a hybrid approach based on MDS and clustering, and STRATSCORE , in terms of requiring fewer random SNPs for inference of population structure. By combining the flexibility and hierarchical nature of phylogenetic trees with the advantage of representing admixture using MDS, our hybrid approach can capture the complex population structures in human populations effectively. Codes can be downloaded from http://people.pcbi.upenn.edu/ approximately lswang/phylostrat/ firstname.lastname@example.com; email@example.com. Supplementary data are available at Bioinformatics online.


Clinical outcomes of minimally invasive endoscopic and conventional sternotomy approaches for atrial septal defect repair.
Concerns remain that minimally invasive atrial septal defect (ASD) repair may compromise patient outcomes. We compared clinical outcomes of adult patients undergoing ASD repair via a minimally invasive endoscopic approach versus a "gold standard" sternotomy. We retrospectively reviewed the clinical outcomes of consecutive patients who underwent ASD patch repair at our institution between 2002 and 2012. We compared in-hospital/30-day mortality, postoperative complications, length of stay in hospital and in the intensive care unit and blood product requirements between patients who underwent right mini-thoracotomy (MT) and those who underwent conventional sternotomy. During the study period, 73 consecutive patients underwent ASD patch repair at our institution: 51 (age 47 ± 16 yr, 66.7% women) in the MT group and 22 (age 46 ± 21 yr, 59.1% women) in the sternotomy group. In-hospital mortality was similar between the 2 groups (MT 0% v. sternotomy 4.5%, p = 0.30). There were no significant differences in any postoperative complications or blood product requirements. No patients in the MT group suffered stroke, retrograde aortic dissection or leg ischemia. Mean intensive care unit (MT 1.2 ± 1.2 d v. sternotomy 1.7 ± 2.2 d, p = 0.26) and hospital length of stays (MT 5.1 ± 2.2 d v. sternotomy 6.3 ± 3.6 d, p = 0.17) were similar between the groups; however, there was a trend toward fewer patients requiring prolonged hospital stays (> 10 d) in the MT group (3.9% v. 18.2%, p = 0.06). Repair of ostium secundum and sinus venosus ASD can be performed safely via MT endoscopic approach with similar outcomes as sternotomy. Patient preference for a more cosmetically appealing incision may be considered without concern of compromised outcomes.


Achievement of safer palladium nanocrystals by enlargement of {100} crystallographic facets.
Developing catalytic and safe nanomaterials is very necessary for the reduction of potential risk to human health; however, this strategy has been found extremely challenging because the enhancement in catalytic activity of nanomaterials is inevitably accompanied with more potent cell injury. The relationship of physicochemical properties and biological responses in catalytic nanomaterials needs to be clarified at the nano-bio interface for achieving the safe application. Herein, high-energy crystallographic facets of palladium (Pd) nanocrystals that have been known to significantly contribute to the catalytic activity were introduced to attenuate the toxicity, and the underlying mechanism was unraveled. Polyhedral Pd nanocrystals with morphology evolution from truncated octahedron to cuboctahedron and cube were prepared for elaborately tuning the extents of high-energy {100} facets, and hierarchical in vitro and in vivo biological evaluation were performed to clarify that Pd nanocrystals exposed with the more {100} facets could show the less toxicity to cells and animals. Density functional theory (DFT) calculation revealed {100} facet exposure was endowed with a strong oxygen adsorption, which weakens the breakage of the water molecule and suppresses the hazardous water dissociation and hydroxyl radical generation, which was supported by electron spin resonance (ESR)-based radical evaluation and X-ray photoelectron spectroscopy (XPS)-based oxygen identification. This means high-energy facet-based catalytic Pd nanocrystals can deliver low toxicity due to their unique surface properties.


Including Family Caregivers In Seriously Ill Veterans' Care: A Mixed-Methods Study.
Family caregivers often serve as unpaid members of the home and community-based care workforce for people with serious illness; as key partners in the home-clinic continuum, they should be included in health care teams. The Campaign for Inclusive Care is an initiative within the Veterans Affairs health care system to improve provider practices for including caregivers of military members in treatment planning and decisions. We defined inclusive care using a literature review, provider interviews, and a caregiver survey. We found that inclusive care involves clear definition of the caregiver role, system policies for inclusion, assessment of caregivers' capacity, explicit involvement of caregivers, and mutuality in caregiver-provider communication. We recommend solutions based on this definition that can inform development of a national caregiver strategy, required of the Department of Health and Human Services by the Recognize, Assist, Include, Support, and Engage Family Caregivers Act of 2018.


Intraoperative monitoring for tethered cord surgery: an update.
Intraoperative neurophysiological recording techniques have found increasing use in neurosurgical practice. The development of new recording techniques feasible while the patient receives a general anesthetic have improved their practical use in a similar way to the use of digital recording, documentation, and video technology. This review intends to provide an update on the techniques used and their validity. Two principal methods are used for intraoperative neurophysiological testing during tethered cord release. Mapping identifies functional neural structures, namely nerve roots, and monitoring provides continuous information on the functional integrity of motor and sensory pathways as well as reflex circuitry. Mapping is performed mostly by using direct electrical stimulation of a structure within the surgical field and recording at a distant site, usually a muscle. Sensory mapping can also be performed with peripheral stimulation and recording within the surgical site. Monitoring of the motor system is achieved with motor evoked potentials. These are evoked by transcranial electrical stimulation and recorded from limb muscles and the external anal sphincter. The presence or absence of muscle responses are the parameters monitored. Sensory potentials evoked by tibial or pudendal nerve stimulation and recorded from the dorsal columns via an epidurally inserted electrode and/or from the scalp as cortical responses are used to access the integrity of sensory pathways. Amplitudes and latencies of these responses are then interpreted. The bulbocavernosus reflex, with stimulation of the pudendal nerve and recording of muscle responses in the external anal sphincter, is used for continuous monitoring of the reflex circuitry. Presence or absence of this response is the pertinent parameter that is monitored. Intraoperative neurophysiology provides a wide and reliable set of techniques for intraoperative identification of neural structures and continuous monitoring of their functional integrity.


The role of innovation and entrepreneurship in promoting common prosperity in China: Empirical evidence from a two-way fixed effects model.
Common prosperity stands as a pivotal concept and objective within China's socialism with distinctive characteristics, serving as a fundamental assurance and basis for ensuring its people's happiness and comprehensive development. This research employs a Coupled Coordination Degree Model to construct a common prosperity Index using data from China between 2010 and 2020. The study investigates the influence of innovation and entrepreneurship on common prosperity while examining the regulating roles played by the government and market during this process. The outcomes demonstrate that innovation substantially facilitates the realization of common prosperity. The relationship between entrepreneurship and common prosperity follows a positive "U"-shaped curve, where entrepreneurship significantly contributes to common prosperity upon reaching a particular scale. Further investigations reveal heterogeneity in the impact of innovation and entrepreneurship on common prosperity. Specifically, innovation significantly contributes to common prosperity in the northern regions, whereas entrepreneurship has a noteworthy impact on common prosperity in the southern regions. Moreover, it is worth noting that both innovation and entrepreneurship have a significant influence on common prosperity in areas characterized by low economic development levels and a scarcity of fixed capital. The fiscal effects of the government attenuate the promoting effect of innovation on common prosperity but enhance the adverse influence of entrepreneurship. On the contrary, market mechanisms mitigate the negative impact of entrepreneurship on common prosperity. Consequently, achieving common prosperity requires strengthened regional innovation cooperation, encouraging advanced regions to lead underdeveloped regions, and leveraging the regulatory roles of both the government and the market, thus progressing gradually towards common prosperity.


Fluorosis of deciduous teeth and first permanent molars in a rural Kenyan community.
The severity and distribution of fluorosis in the deciduous dentition of 76 children in a low-income community near Nairobi were studied. Seventeen children comprised a low-F (fluoride) group (water less than 0.7 ppm F) and 59 a high-F group (water approximately 9 ppm F). The high-F group had scores greater than or equal to 5 in the Thylstrup & Fejerskov classification system for 29% of the deciduous tooth surfaces, compared with 7% in the low-F group. Comparison between the scores of the second deciduous and the first permanent molars showed no significant difference in the high-F group (p greater than 0.001), whereas the deciduous molar was significantly less severely affected in the low-F group (p less than 0.001). The deciduous molars of the two groups differed significantly (p less than 0.002), but not the permanent molars (p greater than 0.10). Early introduction of tea might have been a major contributor to the distributions of fluorosis, particularly in the low-F group.


Protocol for cross-platform characterization of human and murine extracellular vesicles and particles.
Characterization of isolated extracellular vesicles and particles (EVPs) is crucial for determining functions and biomarker potential. Here, we present a protocol to analyze size, number, morphology, and EVP protein cargo and to validate EVP proteins in both humans and mice. We describe steps for nanoparticle tracking analysis, transmission electron microscopy, single-EVP immunodetection, EVP proteomic mass spectrometry and bioinformatic analysis, and EVP protein validation by ExoELISA and western blot analysis. This allows for EVP cross-validation across different platforms. For complete details on the use and execution of this protocol, please refer to Hoshino et al.1.


Norepinephrine regulates calcium signals and fate of oligodendrocyte precursor cells in the mouse cerebral cortex.
Oligodendrocyte precursor cells (OPCs) generate oligodendrocytes, contributing to myelination and myelin repair. OPCs contact axons and respond to neuronal activity, but how the information relayed by the neuronal activity translates into OPC Ca2+ signals, which in turn influence their fate, remains unknown. We generated transgenic mice for concomitant monitoring of OPCs Ca2+ signals and cell fate using 2-photon microscopy in the somatosensory cortex of awake-behaving mice. Ca2+ signals in OPCs mainly occur within processes and confine to Ca2+ microdomains. A subpopulation of OPCs enhances Ca2+ transients while mice engaged in exploratory locomotion. We found that OPCs responsive to locomotion preferentially differentiate into oligodendrocytes, and locomotion-non-responsive OPCs divide. Norepinephrine mediates locomotion-evoked Ca2+ increases in OPCs by activating α1 adrenergic receptors, and chemogenetic activation of OPCs or noradrenergic neurons promotes OPC differentiation. Hence, we uncovered that for fate decisions OPCs integrate Ca2+ signals, and norepinephrine is a potent regulator of OPC fate.


Two-dimensional arrays for medical ultrasound using multilayer flexible circuit interconnection.
The development of 2-D array transducers has received much recent interest. Unfortunately, fabrication of high density 2-D arrays is difficult due to the large number of electrical interconnections which must be made to the back side of the elements. A typical array operating at 2.2 MHz may have 256 or more connections within a 16.4 mm circular footprint. Interconnection becomes even more challenging as operating frequencies increase. To solve this problem, we have developed a multilayer flexible (MLF) circuit interconnect consisting of a polyimide dielectric with inter-laminar vias routing signals vertically and etched metal traces routing signals horizontally. A transducer is fabricated from an MLF by bonding a PZT chip to its surface and dicing the chip into individual elements, with the saw kerf extending partially into the top polyimide layer to ensure physical and electrical isolation of the elements. The KLM model was used to compare the performance of an MLF 2-D array to a conventional hand wired 2-D array. MLF and wire guide transducers were fabricated, each with 256 active elements, 0.4 mm interelement spacing, and 2.2 MHz center frequency. Vector impedance, pulse length, bandwidth, angular response, and cross-coupling were found to be comparable in both types of arrays. Using the MLF, however, fabrication time was reduced dramatically. More importantly, MLF technology may be used to increase 2-D array connection density beyond the limitations of current of hand wired fabrication techniques. Thus MLF circuits provide a means for the interconnection of current and future high frequency 2-D arrays.


Heparin washout in the pediatric Cell Saver bowl.
The possibility of residual heparin in washed red cells transfused to neonatal or pediatric cardiac patients following bypass prompted a measurement of heparin concentrations. Samples were taken during 10 adult and 10 neonatal and pediatric bypass cases. Sample A was from the bypass circuit, Sample B from the Haemonetics Cell Saver bowl inlet before washing, Sample C from the Cell Saver bowl outlet after washing, and Sample D from the patient ten minutes after protamine. Heparin concentrations were measured by a chromogenic assay using activated Factor X. There was no significant difference between the adult and pediatric groups in the levels of heparin concentration on bypass, pre-washing and post-washing, and in the patients following protamine. In the pediatric group, only .002% of the pre-washed heparin remained after washing. This extremely low level of heparin (.0027 units/ml) is only 0.34 units in a 125 ml pediatric unit of Cell Saver blood. Based on post bypass patient samples, this has no clinical significance. Therefore, the Cell Saver can be used safely with neonates and pediatric patients without concern regarding residual heparin when properly processed.


Evaluation of organic anion transporting polypeptide 1B1 and 1B3 humanized mice as a translational model to study the pharmacokinetics of statins.
Organic anion transporting polypeptide (Oatp) 1a/1b knockout and OATP1B1 and -1B3 humanized mouse models are promising tools for studying the roles of these transporters in drug disposition. Detailed characterization of these models will help to better understand their utility for predicting clinical outcomes. To advance this approach, we carried out a comprehensive analysis of these mouse lines by evaluating the compensatory changes in mRNA expression, quantifying the amounts of OATP1B1 and -1B3 protein by liquid chromatography-tandem mass spectrometry, and studying the active uptake in isolated hepatocytes and the pharmacokinetics of some prototypical substrates including statins. Major outcomes from these studies were 1) mostly moderate compensatory changes in only a few genes involved in drug metabolism and disposition, 2) a robust hepatic expression of OATP1B1 and -1B3 proteins in the respective humanized mouse models, and 3) functional activities of the human transporters in hepatocytes isolated from the humanized models with several substrates tested in vitro and with pravastatin in vivo. However, the expression of OATP1B1 and -1B3 in the humanized models did not significantly alter liver or plasma concentrations of rosuvastatin and pitavastatin compared with Oatp1a/1b knockout controls under the conditions used in our studies. Hence, although the humanized OATP1B1 and -1B3 mice showed in vitro and/or in vivo functional activity with some statins, further characterization of these models is required to define their potential use and limitations in the prediction of drug disposition and drug-drug interactions in humans.


[Somatic cell hybridization as 1 of the mechanisms explaining the inadequacy of immunologic surveillance].
The literature analysis has shown that somatic cell hybridization is one of the mechanisms of tumour progression in vivo. This mechanism is suggested to underlie the establishment and development of human tumours, in particular of human breast cancer. Somatic hybridization of cells in human breast cancer is supposed to be one of the mechanisms responsible for the inadequacy of immunologic surveillance. The latter appears to occur in those cases when antigenic determinants of unchanged original cells predominate on the surface of hybrid formations.


An internal ribosome entry site directs translation of the murine gammaherpesvirus 68 MK3 open reading frame.
The gammaherpesviruses characteristically drive the proliferation of latently infected lymphocytes. The murine gammaherpesvirus 68 (MHV-68) MK3 protein contributes to this process in vivo by evading CD8(+)-T-cell recognition during latency, as well as during lytic infection. We analyzed some of the molecular mechanisms that control MK3 expression. No dedicated MK3 mRNA was detected. Instead, the MK3 open reading frame (ORF) was transcribed as part of a bicistronic mRNA, downstream of a previously unidentified ORF, 13M. The 13M/MK3 promoter appeared to extend approximately 1 kb 5' of the transcription start site and included elements both dependent on and independent of the ORF50 lytic transactivator. MK3 was translated from the bicistronic transcript by virtue of an internal ribosome entry site (IRES) element. RNA structure mapping identified two stem-loops between 13M and MK3 that were sufficient for IRES activity in a bicistronic reporter plasmid and a third stem-loop just within the MK3 coding sequence, with a subtler, perhaps regulatory role. Overall, translation of the MHV-68 MK3 bore a striking resemblance to that of the Kaposi's sarcoma-associated herpesvirus vFLIP, suggesting that IRES elements are a common theme of latent gammaherpesvirus immune evasion in proliferating cells.


The Biology of Malignant Mesothelioma and the Relevance of Preclinical Models.
Malignant mesothelioma (MM), especially its more frequent form, malignant pleural mesothelioma (MPM), is a devastating thoracic cancer with limited therapeutic options. Recently, clinical trials that used immunotherapy strategies have yielded promising results, but the benefits are restricted to a limited number of patients. To develop new therapeutic strategies and define predictors of treatment response to existing therapy, better knowledge of the cellular and molecular mechanisms of MM tumors and sound preclinical models are needed. This review aims to provide an overview of our present knowledge and issues on both subjects. MM shows a complex pattern of molecular changes, including genetic, chromosomic, and epigenetic alterations. MM is also a heterogeneous cancer. The recently described molecular classifications for MPM could better consider inter-tumor heterogeneity, while histo-molecular gradients are an interesting way to consider both intra- and inter-tumor heterogeneities. Classical preclinical models are based on use of MM cell lines in culture or implanted in rodents, i.e., xenografts in immunosuppressed mice or isografts in syngeneic rodents to assess the anti-tumor immune response. Recent developments are tumoroids, patient-derived xenografts (PDX), xenografts in humanized mice, and genetically modified mice (GEM) that carry mutations identified in human MM tumor cells. Multicellular tumor spheroids are an interesting in vitro model to reduce animal experimentation; they are more accessible than tumoroids. They could be relevant, especially if they are co-cultured with stromal and immune cells to partially reproduce the human microenvironment. Even if preclinical models have allowed for major advances, they show several limitations: (i) the anatomical and biological tumor microenvironments are incompletely reproduced; (ii) the intra-tumor heterogeneity and immunological contexts are not fully reconstructed; and (iii) the inter-tumor heterogeneity is insufficiently considered. Given that these limitations vary according to the models, preclinical models must be carefully selected depending on the objectives of the experiments. New approaches, such as organ-on-a-chip technologies or in silico biological systems, should be explored in MM research. More pertinent cell models, based on our knowledge on mesothelial carcinogenesis and considering MM heterogeneity, need to be developed. These endeavors are mandatory to implement efficient precision medicine for MM.


Commercial Disinfectants During Disinfection Process Validation: More Failures than Success.
Disinfection process validation is mandatory before introduction of a new disinfectant in hospital services. Commercial disinfection brands often question existing hospital policy claiming greater efficacy and lack of toxicity of their products. Inadvertent inadequate disinfection leads to morbidity, patient's economic burden, and the risk of mortality. To evaluate commercial disinfectants for high, intermediate and low-level disinfection so as to identify utility for our routine situations. This laboratory based experiment was conducted at St Stephen Hospital, Delhi during July-September 2013. Twelve commercial disinfectants: Sanidex®, Sanocid®, Cidex®, SekuSept Aktiv®, BIB Forte®, Alprojet W®, Desnet®, Sanihygiene®, Incidin®, D125®, Lonzagard®, and Glutishield® were tested. Time-kill assay (suspension test) was performed against six indicator bacteria (Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Salmonella Typhi, Bacillus cereus, and Mycobacterium fortuitum). Low and high inoculum (final concentrations 1.5X10(6) and 9X10(6) cfu/ml) of the first five bacteria while only low level of M. fortuitum was tested. Cidex® (2.4% Glutaraldehyde) performed best as high level disinfectant while newer quarternary ammonium compounds (QACs) (Incidin®, D125®, and Lonzagard®) were good at low level disinfection. Sanidex® (0.55% Ortho-pthalaldehyde) though mycobactericidal took 10 minutes for sporicidal activity. Older QAC containing BIB Forte® and Desnet® took 20 minutes to fully inhibit P. aeruginosa. All disinfectants effectively reduced S. Typhi to zero counts within 5 minutes. Cidex® is a good high-level disinfectant while newer QACs (Incidin®, D125®, and Lonzagard®) were capable low-level disinfectants.


Sites, rates, and limits of starch digestion and glucose metabolism in growing cattle.
Growing cattle in the United States consume up to 6 kg of starch daily, mainly from corn or sorghum grain. Total tract apparent digestibility of starch usually ranges from 90 to 100% of starch intake. Ruminal starch digestion ranges from 75 to 80% of starch intake and is not greatly affected by intake over a range of 1 to 5 kg of starch/d. Starch apparently digested in the small intestine decreases from 80 to 34% as starch entering the small intestine increases from 0.2 to 2 kg/d. Starch apparently digested in the large intestine ranges from 44 to 46% of starch entering the large intestine. Approximately 70% of starch digested in the small intestine appears as glucose in the bloodstream. Within the range of starch intakes that do not cause rumen upsets, increasing starch (and energy) intake increases the amount of starch digested in the rumen, increases the supply of starch to the small intestine, increases starch digested in small intestine (albeit at reduced efficiency), and increases starch digested in the large intestine, such that total tract digestibility remains relatively constant. With increased starch intake, most of the starch is still digested in the rumen, but increasing amounts of starch escape ruminal and intestinal digestion, and disappear distal to the ileocecal junction. Again, within the range of starch intakes that do not cause rumen upsets, as starch intake increases, hepatic gluconeogenesis increases, glucose entry increases, and glucose irreversible loss increases, with a significant portion lost as CO2. The ability to increase use of dietary starch to support greater weight gains or improved marbling could come from increasing starch digestion in a healthy rumen or in the small intestine, but we conclude that the main limit to use of dietary starch to support live weight gain is digestion and absorption from the small intestine. Increased oxidation of glucose at greater starch intakes may alter energetic efficiency by sparing other oxidizable substrates, like amino acids.


Voltage clamp of rat and human skeletal muscle: measurements with an improved loose-patch technique.
Intact fibres of human intercostal and rat omohyoid muscles were studied at 23 degree C with a loose-patch voltage-clamp technique that employed two concentric micropipettes to electrically isolate small-diameter (10-15 microns) patches of sarcolemma. This method allows investigation of membrane excitability under highly physiological conditions. Step depolarizations to 0 mV elicited sodium inward currents that reached peak values of up to 20 mA/cm2 within 250 microseconds, and then declined. In human muscle, the reversal potential (ENa) was approximately 40 mV, and maximal conductances (GNa) ranged from 44 to 360 mS/cm2. In rat muscle, ENa was 42 mV and GNa ranged from 100 to 250 mS/cm2. Sodium channels in rat and human muscle were indistinguishable in most aspects of their kinetic behaviour and voltage dependence. Outward potassium currents were small by comparison (usually less than 2 mA/cm2) and saturated at positive potentials. The maximum potassium conductance (GK) ranged from 0 to 19 mS/cm2 (human) and from 4 to 12 mS/cm2 (rat muscle).


[Otoacoustic emission and auditory efferent function testing in normal subjects and patients with sensori-neural hearing loss].
To examine the role of otoacoustic emission and function test of the auditory efferent system in evaluating of auditory functional status in both normal and diseased conditions. Spontaneous and transient evoked otoacoustic emission and efferent function in terms of contralateral white noise induced transient evoked otoacoustic emission suppressions were tested and cross compared in 32 normal ears, 24 ears with cochlear hearing loss and 16 ears with retrocochlear hearing loss. Transient evoked otoacoustic emission amplitude and contralateral suppression, spontaneous otoacoustic emission incidence, peaks and frequency range were significantly reduced in cochlear hearing loss, while in retrocochlear hearing loss transient evoked otoacoustic emission and spontaneous otoacoustic emission levels were significantly higher than in cochlear hearing loss group and showed no suppression. Contralateral acoustic stimulation induced transient evoked otoacoustic emission suppression as an index of efferent function is clinically feasible. Combined otoacoustic emission and efferent tests are of great significance in evaluating cochlea status and auditory central mechanisms. Auditory efferent function is weakened in cochlear lesions and severely damaged in retrocochlear lesions.


Transfer region of pO113 from enterohemorrhagic Escherichia coli: similarity with R64 and identification of a novel plasmid-encoded autotransporter, EpeA.
Enterohemorrhagic Escherichia coli (EHEC) is a prominent, food-borne cause of diarrhea, bloody diarrhea, and the hemolytic uremic syndrome in industrialized countries. Most strains of EHEC carry the locus for enterocyte effacement (LEE) pathogenicity island, but a proportion of isolates from patients with severe disease do not carry LEE and very little is known about virulence factors in these organisms. LEE-negative strains of EHEC typically express Shiga toxin 2 and carry a large plasmid that encodes the production of EHEC hemolysin. In this study, we determined the nucleotide sequence of the transfer region of pO113, the large hemolysin plasmid from LEE-negative EHEC O113:H21 (EH41). This 63.9-kb region showed a high degree of similarity with the transfer region of R64, and pO113 was capable of self-transmission at low frequencies. Unlike R64 and the related dot/icm system of Legionella pneumophila, however, pO113 was unable to mobilize RSF1010. In addition, the pO113 transfer region encoded a novel high-molecular-weight serine protease autotransporter of Enterobacteriaceae (SPATE) protein, termed EpeA. Like other SPATEs, EpeA exhibited protease activity and mucinase activity, but expression was not associated with a cytopathic effect on epithelial cells. Analysis of a second high-molecular-weight secreted protein revealed that pO113 also encodes EspP, a cytopathic SPATE identified previously in EHEC O157:H7. The nucleotide sequences encoding the predicted beta-domains of espP and epeA were identical and also shared significant homology with a third SPATE protein, EspI. Both espP and epeA were detected in several LEE-negative clinical isolates of EHEC and thus may contribute to the pathogenesis of this subset of EHEC.