Clinical application of NGS-based SNP haplotyping for the preimplantation genetic diagnosis of primary open angle glaucoma.
The current study describes a successful case of preimplantation genetic diagnosis (PGD) of primary open angle glaucoma (POAG) and verifies the efficiency of next-generation sequencing (NGS)-based haplotyping for PGD of POAG. In this study, we applied NGS as part of PGD to effectively detect POAG prior to embryo implantation and avoid the prospect of pregnancy termination in event of vertical inheritance of POAG. We used the technique of multiple annealing and looping based amplification cycles (MALBAC) to conduct whole genome amplification (WGA) and to reduce the allele dropout (ADO). We also employed Sanger sequencing to directly detect the mutation c.1109 C > T in MYOC and NGS-based single nucleotide polymorphism (SNP) haplotyping to distinguish the chromosomes that carried the mutation. Copy number variation (CNV) analysis was carried out to determine the copy number of embryos' chromosomes. Of the 4 blastocysts obtained in this study, only 2 (sample 5 and 7) could be successfully amplified by WGA. CNV results indicated that chromosomes of both these samples were balanced (46, XN). Sanger sequencing and NGS-based SNP haplotyping confirmed that sample 7 carried the mutation c.1109 C > T in MYOC, while sample 5 did not. Moreover, no ADO was observed. Thus, blastocyst 5 was transferred into the uterus of the patient, and a healthy baby without the MYOC mutation c. 1109C>T was born 39 weeks after transplantation. Our study suggests that NGS-based SNP haplotyping is an effective technique for the PGD of POAG. Abbreviations: PGD: preimplantation genetic diagnosis; POAG: primary open angle glaucoma; NGS: next-generation sequencing; MALBAC: multiple annealing and looping based amplification cycles chemistry; WGA: whole genome amplification; ADO: allele dropout; SNP: single nucleotide polymorphism; CNV: copy number variation; MYOC: Myocilin; OPTN: Optineurin; WDR36: WD repeat domain 36; CYP1B1: Cytochrome P450 1 B Chain; ICSI: intracytoplasmic sperm injection; TFNA: testicular fine-needle aspiration; TE: trophectoderm; PCR: polymerase chain reaction.


Influence of place of treatment on diagnosis, treatment, and survival in three pediatric solid tumors.
This study examines differences between cancer centers (CC) and noncancer centers (NCC) in terms of management procedures and outcomes for three pediatric solid tumors: Wilms' tumor (N = 147), rhabdomyosarcoma (N = 87), and medulloblastoma (N = 76). Data were derived for the period 1970-1979 from the population-based Greater Delaware Valley Pediatric Tumor Registry maintained at the Children's Cancer Research Center, which routinely collects data on all childhood neoplasms that occur in a 31-county region. Management measures reviewed included the degree to which important pretreatment evaluations were performed, types of therapy used, and extent of follow-up examinations conducted. Outcome variables were three-year disease-free survival and frequency of deaths related to complications of therapy. Differences in three-year disease-free survival between CC and NCC were noted for medulloblastoma (52% v 24%) and rhabdomyosarcoma (48% v 10%), but not for Wilms' tumor (79% v 68%). Among medulloblastoma patients, differences were detected in the frequency of pretreatment evaluations and in the therapy used. The principal management contrast found in rhabdomyosarcoma was that multiagent chemotherapy was used less often in NCC. Wilms' tumor patients were evaluated and treated similarly in the community versus the CC, except for some contrasts in the surgical approach and the frequency of follow-up for the detection of late complications.


MURCS in a male: a further case.
A third case of male with MURCS is described. The case is compared with two others previously reported and it is suggested that ARCS (azoospermia, renal anomaly and cervicothoracic spine disorder) might be a more appropriate acronym.


Internal medicine residents' training in substance use disorders: a survey of the quality of instruction and residents' self-perceived preparedness to diagnose and treat addiction.
Resident physicians are the direct care providers for many patients with addiction. This study assesses residents' self-perceived preparedness to diagnose and treat addiction, measures residents' perceptions of the quality of addictions instruction, and evaluates basic knowledge of addictions. A survey was e-mailed to 184 internal medicine residents at Massachusetts General Hospital in May 2012. Responses were obtained from 55% of residents. Residents estimated that 26% of inpatients they cared for met criteria for a substance use disorder (SUD). Twenty-five percent of residents felt unprepared to diagnose and 62% felt unprepared to treat addiction. Only 13% felt very prepared to diagnose addiction. No residents felt very prepared to treat addiction. Preparedness to diagnose or treat addiction did not differ significantly across postgraduate year (PGY) level. Fifty-five percent rated the overall instruction in addictions as poor or fair. Seventy-two percent of residents rated the quality of addictions training as poor or fair in the outpatient clinical setting, and 56% in the inpatient setting. No resident answered all 6 knowledge questions correctly. Slightly more than half correctly identified the mechanism of buprenorphine and 19% correctly answered a question about naltrexone. Nine percent of residents responded that someone had expressed concern about the respondent's substance use. Despite providing care for a substantial population with addiction, the majority of internal medicine residents in this study feel unprepared to treat SUDs. More than half rate the quality of addictions instruction as fair or poor. Structured and comprehensive addictions curriculum and faculty development are needed to address the deficiencies of the current training system.


Plastidial (p)ppGpp Synthesis by the Ca2+-Dependent RelA-SpoT Homolog Regulates the Adaptation of Chloroplast Gene Expression to Darkness in Arabidopsis.
In bacteria, the hyper-phosphorylated nucleotide, guanosine 3',5'-bis(pyrophosphate) (ppGpp), functions as a secondary messenger under stringent conditions. ppGpp levels are controlled by two distinct enzymes, namely RelA and SpoT, in Escherichia coli. RelA-SpoT homologs (RSHs) are also conserved in plants where they function in the plastids. The model plant Arabidopsis thaliana contains four RSHs: RSH1, RSH2, RSH3 and Ca2+-dependent RSH (CRSH). Genetic characterizations of RSH1, RSH2 and RSH3 were undertaken, which showed that the ppGpp-dependent plastidial stringent response significantly influences plant growth and stress acclimation. However, the physiological significance of CRSH-dependent ppGpp synthesis remains unclear, as no crsh-null mutant has been available. Here, to investigate the function of CRSH, a crsh-knockout mutant of Arabidopsis was constructed using a site-specific gene-editing technique, and its phenotype was characterized. A transient increase in ppGpp was observed for 30 min in the wild type (WT) after the light-to-dark transition, but this increase was not observed in the crsh mutant. Similar analyses were performed with the rsh2-rsh3 double and rsh1-rsh2-rsh3 triple mutants and showed that the transient increments of ppGpp in the mutants were higher than those in the WT. The increase in ppGpp in the WT and rsh2 rsh3 accompanied decrements in the mRNA levels of some plastidial genes transcribed by the plastid-encoded plastid RNA polymerase. These results indicate that the transient increase in ppGpp at night is due to CRSH-dependent ppGpp synthesis and that the ppGpp level is maintained by the hydrolytic activities of RSH1, RSH2 and RSH3 to accustom plastidial gene expression to darkness.


Where do I stand? Examining the effects of leader-member exchange social comparison on employee work behaviors.
Taking an approach integrating principles of leader-member exchange (LMX) differentiation with social comparison theory, we contend that subjective ratings by individuals of their LMX compared to the LMXs of coworkers (labeled LMX social comparison, or LMXSC) explain unique and meaningful variance in outcomes beyond LMX and the actual standing of those individuals in the LMX distribution, referred to as relative LMX, or RLMX. Our findings demonstrate that employees' perceptions of LMXSC are positively related beyond the effects of LMX and RLMX to job performance and citizenship behaviors. Further, we argue that LMXSC mediates the RLMX→outcomes relationships. Analyses showed that, in a sample of 254 employees nested in 50 work groups, a significant part of the effects of RLMX on job performance and citizenship behaviors was mediated through LMXSC after controlling for LMX.


Why do they do it? A grounded theory study of the use of low-value care among primary health care physicians.
The use of low-value care (LVC) is widespread and has an impact on both the use of resources and the quality of care. However, few studies have thus far studied the factors influencing the use of LVC from the perspective of the practitioners themselves. The aim of this study is to understand why physicians within primary care use LVC. Six primary health care centers in the Stockholm Region were purposively selected. Focus group discussions were conducted with physicians (n = 31) working in the centers. The discussions were coded inductively using a grounded theory approach. Three main reasons for performing LVC were identified. Uncertainty and disagreement about what not to do was related to being unaware of the LVC status of a practice, guidelines perceived as conflicting, guidelines perceived to be irrelevant for the target patient population, or a lack of trust in the guidelines. Perceived pressure from others concerned patient pressure, pressure from other physicians, or pressure from the health care system. A desire to do something for the patients was associated with the fact that the visit in itself prompts action, symptoms to relieve, or that patients' emotions need to be reassured. The three reasons are interdependent. Uncertainty and disagreement about what not to do have made it more difficult to handle the pressure from others and to refrain from doing something for the patients. The pressure from others and the desire to do something for the patients enhanced the uncertainty and disagreement about what not to do. Furthermore, the pressure from others influenced the desire to do something for the patients. Three reasons work together to explain primary care physicians' use of LVC: uncertainty and disagreement about what not to do, perceived pressure from others, and the desire to do something for the patients. The reasons may, in turn, be influenced by the health care system, but the decision nevertheless seemed to be up to the individual physician. The findings suggest that the de-implementation of LVC needs to address the three reasons from a systems perspective.


Epidural anesthesia for repeat cesarean delivery in a parturient with Klippel-Feil syndrome.
A patient with Klippel-Feil syndrome, morbid obesity, and scoliosis required cesarean delivery. Her previous cesarean deliveries were performed under general anesthesia. She desired a regional technique. Following aspiration prophylaxis and placement of standard monitors, ultrasound was used to identify midline and L(2-3) interspace. Unintentional dural puncture occurred at 10 cm, with an inability to advance the catheter. On second attempt, an epidural catheter was placed easily. After negative test dose, 18 ml of 2% lidocaine with epinephrine was administered to the patient. A T4 level was achieved. The patient tolerated surgery well. Complete block resolution occurred at 4 hours with no neurologic sequelae.


Effect of sucrose ingestion on plasma glutamate concentrations in humans administered monosodium L-glutamate.
Plasma glutamate concentrations in human subjects are markedly lower when monosodium L-glutamate (MSG) is ingested in consomme with starch than when ingested in consomme alone. This study investigated whether sucrose had a similar effect. Six normal adult subjects (three male, three female) ingested two servings of beef consomme each providing 50 mg MSG/kg body weight in a randomized crossover design. One serving of consomme contained no added carbohydrate; the other provided 0.5 g sucrose/kg body weight. Ingestion of the consomme without sucrose significantly (p less than 0.05) increased the mean plasma glutamate concentration from baseline (4.44 +/- 0.97 mumol/dl) to a peak value of 18.1 +/- 6.99 mumol/dl 30 min after dosing. The area under the plasma glutamate concentration-time curve was 553 +/- 238 mumol/dl X min. When the consomme contained 0.5 g sucrose/kg body weight, both the mean peak plasma glutamate concentration (5.48 +/- 2.19 mumol/dl) and the area under the curve (105 +/- 46 mumol/dl X min) were significantly lower. These data confirm that metabolizable carbohydrate has a significant effect on plasma glutamate concentration response after MSG loading.


[The effect of the vasoconstrictor glycylpressin on the bioavailability of mitomycin C].
During an intra-arterial chemotherapy of patients with liver metastases the plasma concentration-time curves of Mitomycin C (1) after occlusion of the common hepatic artery by the vasoconstricting agent Glycylpressin (2) were investigated. Vasoconstriction by 2 leads to significantly (p less than 0.01) lower blood levels of 1 in contrast to bolus injection of 1 without 2 under identical dosage regimens. This reduction of systemic availability of about 50% leads to a corresponding higher availability for the tumor region.


Expanded utility of the native chemical ligation reaction.
The post-genomic era heralds a multitude of challenges for chemists and biologists alike, with the study of protein functions at the heart of much research. The elucidation of protein structure, localization, stability, post-translational modifications, and protein interactions will steadily unveil the role of each protein and its associated biological function in the cell. The push to develop new technologies has necessitated the integration of various disciplines in science. Consequently, the role of chemistry has never been so profound in the study of biological processes. By combining the strengths of recombinant DNA technology, protein splicing, organic chemistry, and the chemoselective chemistry of native chemical ligation, various strategies have been successfully developed and applied to chemoselectively label proteins, both in vitro and in live cells, with biotin, fluorescent, and other small molecule probes. The site-specific incorporation of molecular entities with unique chemical functionalities in proteins has many potential applications in chemical and biological studies of proteins. In this article, we highlight recent progress of these strategies in several areas related to proteomics and chemical biology, namely, in vitro and in vivo protein biotinylation, protein microarray technologies for large-scale protein analysis, and live-cell bioimaging.


Subcutaneous placement of a free jejunum and pedicled colon segment to create a diversionary conduit for total esophageal reconstruction.
In this report, we describe a case of difficult esophageal reconstruction with a pedicled colon segment interposition and a free jejunal flap. Laryngectomy and bilateral neck dissection for larynx carcinoma had been attempted in a 59-year-old patient 6 years previously. The patient then received radiotherapy. One year later, large resection was performed due to recurrence of the tumor. Since then the patient had been fed through a gastrostomy tube. Previous attempts at esophageal reconstruction in other institutions were unsuccessful. We reconstructed the total esophagus with subcutaneously tunneled pedicled colon segment interposition and a free jejunal flap using the diversionary loop technique to divert the passage of the foot from the pharynx to the new inlet at the buccogingival sulcus, thus keeping the native esophagus untouched. Following a postoperative training period, the patient learned to swallow successfully and smoothly via the new inlet. The patency of the newly reconstructed esophagus was corroborated by radiological imaging. In summary, although the technique requires complex surgical procedures, it is effective and may be considered as an alternative and reliable option in selected cases.


The bioactive lipid 4-hydroxyphenyl retinamide inhibits flavivirus replication.
Dengue virus (DENV), a member of the Flaviviridae family, is a mosquito-borne pathogen and the cause of dengue fever. The increasing prevalence of DENV worldwide heightens the need for an effective vaccine and specific antivirals. Due to the dependence of DENV upon the lipid biosynthetic machinery of the host cell, lipid signaling and metabolism present unique opportunities for inhibiting viral replication. We screened a library of bioactive lipids and modulators of lipid metabolism and identified 4-hydroxyphenyl retinamide (4-HPR) (fenretinide) as an inhibitor of DENV in cell culture. 4-HPR inhibits the steady-state accumulation of viral genomic RNA and reduces viremia when orally administered in a murine model of DENV infection. The molecular target responsible for this antiviral activity is distinct from other known inhibitors of DENV but appears to affect other members of the Flaviviridae, including the West Nile, Modoc, and hepatitis C viruses. Although long-chain ceramides have been implicated in DENV replication, we demonstrate that DENV is insensitive to the perturbation of long-chain ceramides in mammalian cell culture and that the effect of 4-HPR on dihydroceramide homeostasis is separable from its antiviral activity. Likewise, the induction of reactive oxygen species by 4-HPR is not required for the inhibition of DENV. The inhibition of DENV in vivo by 4-HPR, combined with its well-established safety and tolerability in humans, suggests that it may be repurposed as a pan-Flaviviridae antiviral agent. This work also illustrates the utility of bioactive lipid screens for identifying critical interactions of DENV and other viral pathogens with host lipid biosynthesis, metabolism, and signal transduction.


Endoscopic management of upper tract urothelial carcinoma.
While radical nephroureterectomy remains the gold standard of treatment for patients with upper tract urothelial tumors, technological advances have made endoscopic management possible. The careful selection of patients for such an approach is dependent upon an accurate diagnosis and an understanding of the natural history of the disease. High grade tumors behave aggressively and warrant radical extirpation unless an absolute contraindication exists. Motivated patients with low grade tumors and relative contraindications to nephroureterectomy can be managed with percutaneous or retrograde ureteroscopic techniques. High recurrence rates in the ipsilateral upper tract and bladder mandate close surveillance of patients treated conservatively. We review the important diagnostic, staging, technical, and surveillance issues in the endoscopic treatment of upper tract urothelial carcinoma.


Expression of alpha-thymosins in human tissues in normal and abnormal growth.
Radioimmunoassays specific for the N and C termini of human prothymosin alpha and the N terminus of human parathymosin alpha were employed for the measurement of the levels of alpha-thymosins in human thymus, spleen, and liver during normal growth and intestine and breast in malignant growth. A differential expression of the two alpha-thymosins was observed in thymus (prothymosin alpha-rich) and liver (parathymosin alpha-rich). A decline in the levels of both alpha-thymosins was found with age, with prothymosin alpha in thymus showing the sharpest change (15- to 30-fold). The levels of both alpha-thymosins were higher in malignant tissues as compared with healthy ones. In breast cancer, in particular, the mean increase for prothymosin alpha and parathymosin alpha was 17.9- and 11.5-fold, respectively. The major crossreactive material was characterized in all cases as intact prothymosin alpha and parathymosin alpha. These results suggest an in vivo relationship of the expression of alpha-thymosins with the human tissue cell proliferation activity.


Human in vivo-activated CD45R0(+) CD4(+) T cells are susceptible to spontaneous apoptosis that can be inhibited by the chemokine CXCL12 and IL-2, -6, -7, and -15.
The number of T cells that have undergone proliferation after antigen stimulation in vivo must be controlled to prevent excessive accumulation of T cells, autoimmunity, and T cell neoplasia. We describe here that primary human adenotonsillar memory phenotype CD45R0(+) CD4(+) T cells, but not adenotonsillar naive-phenotype CD45RA(+) CD4(+) T cells, or peripheral blood naive or memory CD4(+) T cells, express high levels of activation-associated antigens CD38, CD69, CD71, and HLA-DR. These in vivo-activated CD45R0(+) CD4(+) T cells were susceptible to spontaneous and rapid apoptosis in vitro. Apoptosis could not be inhibited by the disruption of Fas-Fas ligand engagement or by the pan-caspase inhibitor ZVAD. Cross-linking of the T cell antigen receptor did not rescue cells from apoptosis. Apoptosis could be partially inhibited by the chemokine CXCL12/SDF-1, by IL-6, and by the IL-2 receptor common gamma chain-signaling cytokines IL-2, -7, and -15. Inhibitors of phosphatidylinositol 3-kinase accelerated apoptosis. We conclude that after in vivo activation of CD45R0(+) CD4(+) T cells, the cells experience a period of intrinsically elevated sensitivity to apoptosis and that multiple external signals control their survival.


New developments in the understanding and management of persistent pain.
It is proposed that central rather than peripheral factors may be important in pain chronicity. We review recent empirical findings on these processes and discuss implications for treatment and prevention. The literature on neuroimaging of pain and on learning processes shows that learning-induced functional and structural brain changes involving sensorimotor, as well as limbic and frontal, areas are important in the transition from acute to chronic pain. These alterations share many similarities with brain changes in emotional disorders and the specificity for pain needs to be determined. Further important contributors to chronic pain may be disturbed processing of the body image, impaired multisensory integration and faulty feedback from interoceptive processes. These findings have led to new treatment approaches that focus on the extinction of aversive memories, restoration of the body image and normal brain function and include approaches such as brain stimulation, mirror training, virtual reality applications or behavioral extinction training. We propose that chronic pain is characterized by learning-related and memory-related plastic changes of the central nervous system with concomitant maladaptive changes in body perception. These alterations require new treatments that focus on the alteration of central pain memories and maladaptive body perception.


Search for picornaviruses at onset of inflammatory myopathy.
Picornaviruses may not play a role as persistent agents in the inflammatory myopathies, but it is still thought likely that they may act as triggers of an autoimmune process. Forty one muscle biopsy specimens, taken from three weeks to six months (mean four months) after onset, were examined using three different picornaviral primers and PCR. Moderate to severe disease activity was evident in all specimens. The results were compared with those of 18 biopsy specimens examined later in the disease course, and with specimens from 27 patients with non-inflammatory myopathies. All results were negative. Thus, even as early as three weeks after clinical disease appears, picornaviruses are not detectable in these disorders.


The measurement and control of high-risk host cell proteins for polysorbate degradation in biologics formulation.
Nonionic surfactant polysorbates, including PS-80 and PS-20, are commonly used in the formulation of biotherapeutic products for both preventing surface adsorption and acting as stabilizer against protein aggregation. Trace levels of residual host cell proteins (HCPs) with lipase or esterase enzymatic activity have been shown to degrade polysorbates in biologics formulation. The measurement and control of these low abundance, high-risk HCPs for polysorbate degradation are an industry-wide challenge to achieve desired shelf life of biopharmaceuticals in liquid formulation, especially for high-concentration formulation product development. Here, we reviewed the challenges, recent advances, and future opportunities of analytical method development, risk assessment, and control strategies for polysorbate degradation during formulation development with a focus on enzymatic degradation. Continued efforts to advance our understanding of polysorbate degradation in biologics formulation will help develop high-quality medicines for patients.


[Laparoscopic sigmoidectomy].
The laparoscopic approach has emerged as an alternative to trephine stoma construction. In the present series, eight patients underwent laparoscopic faecal diversion. We performed two endsigmoidostomies and six sigmoid loop ostomies. Indications for diversion included inoperable obstructing pelvic carcinoma in three patients, perianal fistulas in another three--and faecal incontinence in one patient. A temporary loop-sigmoidostomy was performed in one patient after anal sphincteroplasty. Three or four ports were used. After mobilisation, the sigmoid colon was pulled through the abdominal wall. When an end-sigmoidostomy was performed the bowel was transected intra-abdominally. There were no operative complications and all stomas functioned early and well. Laparoscopic sigmoidostomy is safe and allows intra-abdominal exploration and mobilisation of the colon without the inconvenience of a mid-line surgical wound.


Wearable Tendon Kinetics.
This study introduces a noninvasive wearable system for investigating tendon loading patterns during outdoor locomotion on variable terrain. The system leverages shear wave tensiometry, which is a new approach for assessing tendon load by tracking wave speed within the tissue. Our wearable tensiometry system uses a battery-operated piezoelectric actuator to induce micron-scale shear waves in a tendon. A data logger monitors wave propagation by recording from two miniature accelerometers mounted on the skin above the tendon. Wave speed is determined from the wave travel time between accelerometers. The wearable system was used to record Achilles tendon wave speed at 100 Hz during 1-km outdoor walking trials in nine young adults. Inertial measurement units (IMUs) simultaneously monitored participant position, walking speed, and ground incline. An analysis of 5108 walking strides revealed the coupled biomechanical effects of terrain slope and walking speed on tendon loading. Uphill slopes increased the tendon wave speed during push-off, whereas downhill slopes increased tendon wave speeds during early stance braking. Walking speed significantly modulated peak tendon wave speed on uphill slopes but had less influence on downhill slopes. Walking speed consistently induced greater early stance wave speeds for all slopes. These observations demonstrate that wearable shear wave tensiometry holds promise for evaluating tendon tissue kinetics in natural environments and uncontrolled movements. There are numerous practical applications of wearable tensiometry spanning orthopedics, athletics, rehabilitation, and ergonomics.


GSK-3β-mediated fatty acid synthesis enhances epithelial to mesenchymal transition of TLR4-activated colorectal cancer cells through regulation of TAp63.
Glycogen synthase kinase-3β (GSK-3β) in cancer cells is a critical regulatory component of both cellular metabolism and epithelial-mesenchymal transition (EMT) processes via regulation of the β-catenin/E-cadherin and phosphoinositide 3-kinase (PI3K)/AKT signaling pathway. Lipogenesis of cancer cells also plays a critical role in survival and metastasis. We investigated the role of GSK-3β-mediated intracellular fatty acid synthesis to control EMT in TLR4-activated colorectal cancer cells and the underlying regulatory mechanism. Engagement of TLR4 with lipopolysaccharide (LPS) in colon cancer cells promoted the induction of phosphorylated GSK-3β and related lipogenic enzymes as well as the expression of CD74, CD44 and macrophage inhibitory factor (MIF), but decreased expression of transcriptionally active p63 (TAp63). In addition, targeted inhibition of GSK-3β using SB216763 was accompanied by decreased intracellular fatty acid synthesis and blockage of CD74 and CD44 expression, whereas it reversed the level of TAp63. Although TAp63 overexpression had no effect on the expression of CD74 and CD44 in LPS-treated colon cancer cells, GSK-3β-dependent fatty acid synthesis and invasive activity were significantly suppressed. Notably, inhibition of CD44 or CD74 by siRNA not only attenuated de novo lipogenesis and migratory activity but also restored the expression of TAp63 in LPS-activated colon cancer cells. These results suggest that TAp63-mediated GSK-3β activation induced by TLR4 stimulation triggers migration and invasion of colon cancer cells through the regulation of lipid synthesis and GSK-3β-mediated CD74/CD44 expression could be a target to control fatty acid-related EMT process through the modulation of TAp63 expression.


Risk factors for Plasmodium falciparum hyperparasitaemia in malarious children.
Hyperparasitaemia is a feature of childhood severe malaria but there is little information on the risk factors for hyperparasitaemia in malarious children The risk factors associated with Plasmodium falciparum hyperparasitaemia, defined as asexual parasitaemia > 250,000/μl, at presentation were evaluated in 3338 malarious children enrolled prospectively between 2008 and 2010 in an endemic area of southwestern Nigeria. At enrolment, 97 (3%) of 3338 malarious children had hyperparasitaemia. In a multiple regression model, 3 factors were found to be independent risk factors for the presence of hyperparasitaemia at enrolment: an age ≤ 11 years (Adjusted odds ratio [AOR] = 2.85, 95% confidence interval [CI] 1.23-6.61, P = 0.014), fever (AOR = 2.02, 95% CI 1.23-3.29, P = 0.005), and enrolment after year 2008 (AOR = 0.42, 95% CI 0.24-0.73, P = 0.002). Duration of illness ≤ 3 d was associated with increased risk of hyperparasitaemia. There was no association between season and hyperparasitaemia. Compared to non-hyperparasitaemia, hyperparasitaemia was associated with an increased risk of progression to cerebral malaria (P < 0.0001). The risk of progression in hyperparasitaemic children was higher in < 5-year olds (P = 0.02). Young age and presence of fever are independent risk factors for hyperparasitaemia which is associated with an increased risk of progression to cerebral malaria. The findings have implications for case and community management of childhood hyperparasitaemia and for malaria control efforts in sub-Saharan Africa where severe malaria is relatively common.


Stereoselective synthesis of 2,4,5-trisubstituted piperidines by carbonyl ene and Prins cyclisations.
An approach to 2,4,5-trisubstituted piperidines is reported, in which the key step is the Prins or carbonyl ene cyclisation of aldehydes of the type 1. Prins cyclisation catalysed by concentrated hydrochloric acid in CH(2)Cl(2) at -78 degrees C afforded good yields of two of the four possible diastereomeric piperidines, with the 4,5-cis product 7 predominating in a diastereomeric ratio of up to 94:6. The diastereoselectivity of the cyclisation decreased as the 2-substituent increased in size, becoming unselective for very bulky 2-substituents. In contrast, cyclisation catalysed by MeAlCl(2) in CH(2)Cl(2) or CHCl(3) at temperatures of between 20-60 degrees C, favoured the 4,5-trans diastereomer 8, in a diastereomeric ratio of up to 99:1. The low-temperature cyclisations catalysed by HCl proceed under kinetic control via a mechanism involving the development of significant carbocationic character, in which the 4,5-cis cation is more stable than the 4,5-trans cation as a result of overlap with the neighbouring oxygen. The cyclisations catalysed by MeAlCl(2) proceed under thermodynamic control, affording the product in which both the 4- and 5-substituents are equatorial.


Directed self-assembly at the 10 nm scale by using capillary force-induced nanocohesion.
We demonstrated a new nanoassembly strategy based on capillary force-induced cohesion of high-aspect ratio nanostructures made by electron-beam lithography. Using this strategy, ordered complex pattern were fabricated from individual nanostructures at the 10 nm length scale. This method enables the formation of complex designed networks from a sparse array of nanostructures, suggesting a number of potential applications in fabrication of nanodevices, nanopatterning, and fluid-flow investigations.


Antinociceptive effects of matrine on neuropathic pain induced by chronic constriction injury.
Sophora alopecuroides L. (Leguminosae) is a commonly used Chinese herbal drug that possesses antipyretic, anti-inflammatory and analgesic effects. Among various alkaloids isolated from S. alopecuroides, matrine has been identified as the major bioactive component contributing to a variety of pharmacological effects, and studies have also shown that matrine has an analgesic effect. To investigate the antinociceptive effects of matrine on neuropathic pain induced by chronic constriction injury (CCI) in mice. The von Frey, plantar, cold-plate, locomotor activity and rota-rod test were performed to assess the degree of mechanical, radiant, thermal, spontaneous locomotor activity and motor coordination changes respectively, at different time intervals, i.e., one day before surgery and 7, 8, 10, 12 and 14 days post surgery. Matrine was administered from the 8th day after the surgery for seven days. Our present study shows that matrine at the dose of 30 mg/kg i.p. increased the paw withdrawal threshold (0.88 ± 0.16), paw withdrawal latency (7.01 ± 0.11) and the counts of paw withdrawal (19.7 ± 1.15) from the day 8 for the nerve injured paw compared to the CCI group (0.18 ± 0.04, 4.62 ± 0.18, 44.3 ± 2.99, respectively). Matrine, in a dose-dependent effect, was also found to produce a protective role in both plantar and cold-plate tests. The analysis of the effect supports the hypothesis that matrine is useful in neuropathic pain therapy. The results of this study suggest that matrine could be useful in the treatment of different kinds of neuropathic pains as an adjuvant to conventional medicines.


Estimating the impact of mandatory folic acid fortification on the folic acid intake of Australian women of childbearing age.
The primary aim of this study was to estimate the impact of mandatory folic acid (FA) fortification of bread-making flour on the FA intake of Australian women of childbearing age (16-44 years). The secondary objective was to investigate the relationship between estimated FA intake and socio-economic status (SES) and age. Dietary modelling was used to estimate FA intake under four mandatory fortification scenarios--no supplement use, supplement use unrelated to FA intake, supplement use only among the highest consumers of bread, and increased supplement use. Data were obtained from the 1995 National Nutrition Survey for food intake patterns, the 2007 Victorian Population Health Survey for FA supplement use, and a marketplace survey. It is estimated that the National Health and Medical Research Council (NHMRC) recommendation for an additional 400 μg/day [DOSAGE ERROR CORRECTED] FA will be achieved by a minimum of 3.9, 25.4, 21.7 and 30% of the target population under scenarios 1-4, respectively. The FA upper level of intake is exceeded by a maximum of 0.1, 1.7, 6.1 and 4.1% of the target population for scenarios 1-4, respectively. Conclusions : Mandatory FA fortification is not sufficient for the NHMRC recommendations for minimum and maximum intakes to be met by all of the target population under a number of plausible behaviour scenarios. Targeted nutrition education campaigns are needed for SES and age sub-groups and research of this nature should be extended to other population groups. Monitoring and evaluation of this policy will be important to ensure appropriate FA intake.


The effect of donor age on survival after lung transplantation.
Historically, donor age above 55 years has been considered to be a relative contraindication for organ transplantation. The shortage of organs for transplantation has led to the expansion of the donor pool by accepling older donors. To compare the 1 year follow-up in patients after lung transplantation from older donors (> 50 years old) and in patients after transplantation from younger donors (< or = 50 years). The study group comprised all adult patients who underwent lung transplantation at the Rabin Medical Center between May 1997 and August 2001. Donors were classified into two groups according to their age: < or = 50 years (n = 20) and > 50 years (n = 9). Survival, number and total days of hospitalization, development of bronchiolitis obliterans syndrome, and pulmonary function tests, were examined 1 year after transplantation. We performed 29 lung transplantations in our center during the observed period. Donor age had no statistically significant impact on 1 year survival after lung transplantation. There was no statistically significant effect on lung function parameters, the incidence of hospitalization or the incidence of bronchiolitis obliterans between both donor age groups at 1 year after transplantation. Donor age did not influence survival or important secondary end-points 1 year after lung transplantation By liberalizing donor criteria of age up to 65 years, we can expand the donor pool, while assessing other possible mechanisms to increase donor availability.


Effect of 6 Months of Physical Training on the Physical Fitness of Young Brazilian Army Cadets.
Following the increase in the employment of women in conflicts around the world, the federal government of Brazil enacted a law which determines the participation of women in the military. The aim of this study was to analyze the effect of six months of physical training (PT) on the physical fitness of young Brazilian Army cadets to carry out the physical assessments provided in military training. Sixty-eight members of the (19.4 ± 1.0 years) military from the Brazilian Army (BA), with BMI of (23.61 ± 2.17/21.81 ± 2.26) respectively and divided in two groups (men/women) participated in the study. PT was conducted by Manual EB20-MC10.350. Anthropometric measurements and assessment of body composition by dual X-ray absorptiometry were performed. The Student's t test, percentage evolution equation, and Levene test were used. Results showedasignificant increase in anthropometric variables and cardiorespiratory fitness in both groups. Bone health variables and visceral fat presented a significant increase in the malegroup. In terms of muscle fitness handgrip and isometric strength there was no significant variation between the groups and push-up and pull-up there was significant variation between the groups. Percentage evolution was greater in female group. The conclusion shows PT was able to cause beneficial changes, promoting positive improvement in bone health, especially in women. Also, PT was shown to enhance cardiorespiratory capacity, and muscle fitness of the upper limbs in all participants.


Liver transplantation following hepatic artery avulsion in a trauma patient.
Hepatic artery avulsion following politrauma is an extremely rare condition with a very high mortality rate. Management is based on damage control surgery given the precarious situation of these patients. Ligating the artery is one option under such circumstances, despite potential consequences including ischemic cholangiopathy (IC). Ischemic cholangiopathy, which can be caused by an insufficient blood supply to the bile duct, generally results in stricture and recurrent cholangitis, and the need for a liver transplant in extreme cases. We present the case of a 37-year-old male with multiple traumas after falling from the third floor of a building. He was hemodynamically unstable upon arrival at the emergencies department, with no improvement on administration of aggressive fluid therapy. A Echo-FAST exam evidenced fluid in all quadrants, so the patient was transferred to the operating room where a 4-litre hemoperitoneum secondary to total avulsion of the proper hepatic artery was observed. The patient required massive transfusion and vasoactive drugs, with instability throughout the intervention; therefore, we decided to ligate the proper hepatic artery. Hepatic dysfunction and diffuse IC with multiple episodes of recurrent cholangitis were observed during the postoperative period. Given the irreversible clinical picture, we opted for a liver transplant 70 days after the patient's initial admission. The patient died on Day 34 post-transplant due to irreversible ischemic brain damage and a right occipital hemorrhage. Hepatic artery avulsion due to trauma is very rare and its management very complex, and in certain situations the artery must be ligated. The main consequence of ligating the hepatic artery is IC, which is more frequently observed secondary to iatrogenic lesions or systemic diseases, while very few cases have been published in which IC is secondary to hepatic artery avulsion caused by hepatic trauma. Treatment depends on the extent of ischemia, and when the damage is diffuse, as in our case, it may involve a liver transplant.


New variables reported to the UNOS registry and their impact on cadaveric renal transplant outcomes - a preliminary study.
Several new factors collected by UNOS since 1994 were examined with regard to their effects on early graft function and 6-month graft survival in this study. Medicare patients had a lower 6-month graft survival rate than private insurance patients, though the difference was not statistically significant. Medicaid patients had a significantly lower graft survival rate than private insurance patients (p=0.04). Blacks had the same graft survival as Whites when they had comparable coverage. Both Blacks and Whites had higher graft survival with private insurance and lower survival with government funding. Non-heartbeating cadaver-donor kidneys yielded a lower graft survival rate than heartbeating donors (p=0.03). Experience with non-heartbeating donors is just beginning with only 89 kidneys compared with 8,766 heartbeating donor kidneys. Kidneys from donors with hypertension resulted in lower graft survival rates than those from donors without hypertension (p=0.01). In these preliminary studies, donor history of diabetes, smoking, alcohol dependency, I.V. drug use and cancer did not have a significant impact on graft survival. Patients who had a high body mass index (>26) had a lower 6-month graft survival rate (p=0.009). Patients who received kidneys from cadaver donors with a low body mass index (<20) had a lower 6-month graft survival rate (p=0.02). When the recipient was more than 30 kg heavier than the donor, a lower 6-month graft survival rate was obtained (p=0.007). When the weight ratio was greater than 2.0, again a lower survival was noted (p<0.001). A recipient-to-donor height ratio of more than 1.25 resulted in an 80% 6-month graft survival which was significantly less than the 87% survival rate among those with a ratio less than 1.25 (p<0.001). This cut-off was found to identify patients with the lowest 6-month survival rate, suggesting that donor-recipient pairs with a height ratio above 1.25 should not be transplanted. A 6-month graft survival rate of 90% was reported with a SMI of 16-20, where SMI = weight of recipient divided by the square of the donor height. From this preliminary study, the SMI appears to be the best predictor of high graft survival since 1,450 patients were in this category with a 90% survival at 6 months.


Physical-chemical properties of furosemide nanocrystals developed using rotation revolution mixer.
Recently, several approaches have been reported to improve the dissolution rate and bioavailability of furosemide, a class IV drug. However, to the best of our knowledge, none of them proposed nanocrystals. In the last decade, nanocrystals successfully addressed solubility issues by increasing surface area and saturation solubility, both leading to an increase in the dissolution rate of poor water soluble drugs. The preparation of furosemide nanocrystals was by a rotation revolution mixer method. Size distribution and morphology were performed using laser diffraction and scanning electron microscopy, respectively. In addition, differential scanning calorimetry, thermogravimetry, X-ray powder diffraction (XRD) and low frequency shift-Raman spectroscopy allowed investigating the thermal properties and crystalline state. Solubility saturation and intrinsic dissolution rate (IDR) studies were conducted. The thermal analysis revealed lower melting range for the nanocrystals comparing to furosemide. Moreover, a slight crystalline structure change to the amorphous state was observed by XRD and confirmed by low frequency shift Raman. The particle size was reduced to 231 nm with a polydispersity index of 0.232, a 30-fold reduction from the original powder. Finally, the saturation solubility and IDR showed a significant increase. Furosemide nanocrystals showed potential for development of innovative formulations as an alternative to the commercial products.


Combined treatment with chenodeoxycholic acid and pravastatin improves plasma cholestanol levels associated with marked regression of tendon xanthomas in cerebrotendinous xanthomatosis.
We studied the effect of chenodeoxycholic acid (CDCA) and a competitive HMG-CoA reductase inhibitor, pravastatin, on clinical symptoms and sterol metabolism in a 36-year-old Japanese man with cerebrotendinous xanthomatosis (CTX). He had marked tendon xanthomas and mild dementia, with obvious electroencephalographic (EEG) abnormalities. He was treated for 2 years with CDCA alone (0.6 g/d) and then for a further year with the combination of pravastatin (10 mg/d) and CDCA (0.6 g/d). For the following year, he was given pravastatin alone, and then was returned to combined treatment again. The plasma cholestanol level before treatment was 3.12 mg/dL, which was 20 times above the control level. After CDCA alone, the plasma cholestanol was reduced to 1.96 mg/dL, and this was further reduced to 0.92 mg/dL by combination therapy with CDCA and pravastatin. However, after the discontinuation of CDCA, his cholestanol levels returned to the pretreatment levels despite the continuing of pravastatin treatment. When the combination therapy was restarted, his cholestanol level was once again markedly reduced. His clinical symptoms showed a close association with the plasma cholestanol level; the xanthomas regressed remarkably and the mental retardation improved in association with normalization of EEG findings during treatment with CDCA alone or in combination with pravastatin. However, during treatment with pravastatin alone, his tendon xanthomas enlarged again and slow waves reappeared on the EEG. Because inhibition of cholesterol synthesis by treatment with the HMG-CoA reductase inhibitor alone was not effective in causing a reduction of cholestanol, the increase in plasma cholestanol levels in CTX may not have been solely due to increased cholesterol synthesis.(ABSTRACT TRUNCATED AT 250 WORDS)


Four-wavelength laser based on intracavity BaWO₄ Raman conversions of a dual-wavelength Q-switched Nd:YLF laser.
By using diode-end-pumped acousto-optically Q-switched intracavity Raman laser configurations, we demonstrate a four-wavelength laser emitting at 1047.0, 1053.0, 1159.4 and 1166.8 nm. Two Nd:YLiF₄ crystals are employed to generate 1047.0-nm and 1053.0-nm laser radiations. These two lasers are then frequency converted by a BaWO₄ Raman crystal to generate 1159.4-nm and 1166.8-nm first-Stokes waves. With pulse synchronization realized, we obtain the maximum output powers of 427, 418, 423 and 332 mW for 1047.0-nm, 1053.0-nm, 1159.4-nm and 1166.8-nm lasers, respectively. The total optical-to-optical conversion efficiency is 15.1%.


Lysosomal-associated transmembrane protein 5 deficiency exacerbates cerebral ischemia/reperfusion injury.
Lysosomal-associated transmembrane protein 5 (LAPTM5) has been demonstrated to be involved in regulating immunity, inflammation, cell death, and autophagy in the pathophysiological processes of many diseases. However, the function of LAPTM5 in cerebral ischemia-reperfusion (I/R) injury has not yet been reported. In this study, we found that LAPTM5 expression was dramatically decreased during cerebral I/R injury both in vivo and in vitro. LAPTM5 knockout (KO) mice were compared with a control, and they showed a larger infarct size and more serious neurological dysfunction after transient middle cerebral artery occlusion (tMCAO) treatment. In addition, inflammatory response and apoptosis were exacerbated in these processes. Furthermore, gain- and loss-of-function investigations in an in vitro model revealed that neuronal inflammation and apoptosis were aggravated by LAPTM5 knockdown but mitigated by its overexpression. Mechanistically, combined RNA sequencing and experimental verification showed that the apoptosis signal-regulating kinase 1 (ASK1)-c-Jun N-terminal kinase (JNK)/p38 pathway was mainly involved in the detrimental effects of LAPTM5 deficiency following I/R injury. Specifically, LAPTM5 directly interacts with ASK1, leading to decreased ASK1 N-terminal dimerization and the subsequent reduced activation of downstream JNK/p38 signaling. In conclusion, LAPTM5 was demonstrated to be a novel modulator in the pathophysiology of brain I/R injury, and targeting LAPTM5 may be feasible as a stroke treatment.


miR-10b promotes migration and invasion in nasopharyngeal carcinoma cells.
MicroRNA-10b (miR-10b) has been reported to play an important role in some types of cancer, but the effects and possible mechanisms of action of miR-10b in the metastasis of nasopharyngeal carcinoma cells (NPC) have not been explored. The aim of the present study was to investigate the function of miR-10b in nasopharyngeal carcinoma and to determine the molecular mechanisms underlying its action. The MTT assay was used to assess proliferation of CNE-2Z cells. Wound healing and transwell migration assays were applied to assess cell migration and invasion, while and expression of E-cadherin and MMP-9 were detected using Western blot analysis. Real-time PCR was employed to detect the expression of genes related to migration and invasion and the 2-??Ct method was used to calculate the degree of expression. MTT assay showed the expression of miR-10b to have no effect on the proliferation of NPC cell lines. The wound healing assay showed that miR-10b mimics promoted the mobility and invasion of NPC cell lines. Inhibitors of miR-10b reduced the ability of NPC cell lines to migrate and invade. In addition, the expression of genes related to migration and invasion, such as E-cadherin, vimentin, and MMP-9, were confirmed to be different in the CNE-2Z NPC cell line transfected with miR-10b mimics and with miR-10b inhibitors. In the present study, miR-10b was found to upregulate the expression of MMP-9 and knockdown of miR-10b was found to significantly downregulate the expression of E-cadherin. On the whole, these results showed that miR-10b plays an important role in the invasion and metastasis of NPC cells.


Design, development and optimization of a novel time and pH-dependent colon targeted drug delivery system.
The aim of present study was to develop a time- and pH-dependent system for delivering mesalamine to the colon. The system consists of the core tablet of mesalamine which is compression coated with hydroxypropyl methylcellulose (HPMC K4M) (time-dependent factor). This is then coated with pH-dependent polymer Eudragit L100. The simplex lattice design was adopted to optimize the independent variables i.e. amount of HPMC (X1), dextrose (X2) and polyvinyl pyrollidone (PVP) (X3) and to study their effect on the dependent variables i.e. lag time and time for 50% drug dissolution (t50). The results of the linear interactive model and graphical representation revealed that as the amount of HPMC increases, the lag time and t50 value also increases and as the amount of dextrose and PVP were increased the lag time and t50 value decreases.


Effect of pindolol, a beta-adrenoceptor blocking agent, on lipid metabolism.
Pindolol, a beta-adrenoceptor blocking agent, was administered to rats to determine the effects of the drug on lipid and lipoprotein in plasma and tissues. The drug caused significant reductions in plasma triglyceride, free fatty acid and very low density lipoprotein. As for lipoprotein lipase activity, the drug produced varying effects; an increase in the heart and a reduction in the adipose tissue. No significant changes in plasma cholesterol, low density lipoprotein and high density lipoprotein cholesterol were observed. The disturbance of growth was noted following pindolol treatment.


Mechanisms involved in the inhibitory effect of chronic alcohol exposure on pancreatic acinar thiamin uptake.
Pancreatic acinar cells (PAC) obtain thiamin from the circulation via a carrier-mediated process that involves thiamin transporters 1 and 2 (THTR-1 and THTR-2; products of SLC19A2 and SLC19A3, respectively). Chronic alcohol exposure of PAC inhibits thiamin uptake, and, on the basis of in vitro studies, this inhibition appears to be transcriptionally mediated. The aim of this study was to confirm the involvement of a transcriptional mechanism in mediating the chronic alcohol effect in in vivo settings and to delineate the molecular mechanisms involved. Using transgenic mice carrying full-length SLC19A2 and SLC19A3 promoters, we found that chronic alcohol feeding led to a significant reduction in the activity of SLC19A2 and SLC19A3 promoters (as well as in thiamin uptake and expression of THTR-1 and -2). Similar findings were seen in 266-6 cells chronically exposed to alcohol in vitro. In the latter studies, the alcohol inhibitory effect was found to be mediated via the minimal SLC19A2 and SLC19A3 promoters and involved the cis-regulatory elements stimulating protein 1 (SP1)/gut-enriched Kruppel-like factor and SP1-GG-box and SP1/GC, respectively. Chronic alcohol exposure of PAC also led to a significant reduction in the expression of the SP1 transcription factor, which upon correction (via expression) led to the prevention of alcohol inhibitory effects on not only the activity of SLC19A2 and SLC19A3 promoters but also on the expression of THTR-1 and -2 mRNA and thiamin uptake. These results demonstrate that the inhibitory effect of chronic alcohol exposure on physiological/molecular parameters of thiamin uptake by PAC is mediated via specific cis-regulatory elements in SLC19A2 and SLC19A3 minimal promoters.


A Promising Role of TGF-β Pathway in Response to Regorafenib in Metastatic Colorectal Cancer: A Case Report.
Colorectal cancer (CRC) is one of the most common cancer types around the world. The prognosis of patients with advanced diseases is still poor in spite of currently available therapeutic options. Regorafenib is an oral tyrosine kinase inhibitor (TKI) approved to treat refractory metastatic colorectal cancer (mCRC). We investigated Somatic mutations in several genes involved in immunological response and cancer progression in both long/short responder mCRC patients who underwent third-line therapy with regorafenib to identify predictive biomarkers of response using Ion Torrent PGM sequencing and bioinformatic tools. We found Somatic mutations in TGFBR1, TGFBR2, and TGFBR3 genes in primary tumor and metastases samples of long-responder patients. Furthermore, our bioinformatic results show that they were mainly enriched in immune response, cell junction, and cell adhesion in long responder patients, particularly in primary tumor and metastatic sites. These data suggest that the TGF-b pattern could be the leading actor of a prolonged response to this drug.


A national audit of the management pathways for new HIV diagnoses.
The British HIV Association recommends that new diagnoses be reviewed by an HIV specialist within two weeks. NHS England outcome measures include the proportion of new diagnoses commencing antiretroviral therapy (ART) within 91 days. We aimed to review the extent to which these recommendations were followed, to explore the topics discussed with new diagnoses, and to identify reasons behind delayed ART initiation. UK specialist HIV services were invited to retrospectively review the notes of their last 40 new diagnoses over a 15-month period. One-hundred and thirty-two services provided data for 2281 eligible individuals. Most new diagnoses were reviewed by a specialist within two weeks (67.7%) and were commenced on ART within 91 days (83%), however, there were some concerning delays in those tested at home and in general practice. Partner notification and treatment benefits were discussed with most individuals, unlike the availability of community support and U = U ("undetectable equals untransmittable"). Lengthy delays in ART initiation were mostly due to individuals initially declining ART or missing appointments. Our findings suggest a need for more streamlined pathways into HIV care, review of new diagnoses who have not commenced ART within 8 weeks, and protocol development to ensure discussion of relevant topics.


Adverse childhood experiences of persons at risk for Huntington's disease or BRCA1/2 hereditary breast/ovarian cancer.
Huntington's disease (HD) is known to have a negative impact on family life. Offspring of HD patients may be exposed to adversity in childhood because of the parent's disease and its psychological consequences. BRCA1/2 hereditary breast and ovarian cancer (BRCA1/2) increases the risk for offspring of being exposed to parental disease or loss. Childhood adversity is associated with psychopathology and various other problems in later life. Adverse childhood experiences (ACEs) before age 16 were assessed in adults at 50% risk for HD (n = 74) or BRCA1/2 (n = 82) and in controls (n = 101), using the Negative Life Events Scale. Mean number and occurrence of ACEs were compared between groups. The odds of having experienced adversity in childhood were higher in HD offspring and BRCA1/2 offspring than in controls. HD offspring reported a higher mean number of ACEs than controls or BRCA1/2 offspring. In HD offspring, the prevalence of parental disease and parental dysfunction experienced before age 16 was higher than in controls. In BRCA1/2 offspring, the prevalence of parental loss before age 16 was higher than in controls. This study indicates that 53% of HD offspring and 45% of BRCA1/2 offspring are exposed to adversity in childhood or adolescence. The relevance of these findings for counseling in predictive testing programs, reproductive decision-making, and child rearing matters is discussed.


Implicit perceptual anticipation triggered by statistical learning.
Our environments are highly regular in terms of when and where objects appear relative to each other. Statistical learning allows us to extract and represent these regularities, but how this knowledge is used by the brain during ongoing perception is unclear. We used rapid event-related fMRI to measure hemodynamic responses to individual visual images in a continuous stream that contained sequential contingencies. Sixteen human observers encountered these statistical regularities while performing an unrelated cognitive task, and were unaware of their existence. Nevertheless, the right anterior hippocampus showed greater hemodynamic responses to predictive stimuli, providing evidence for implicit anticipation as a consequence of unsupervised statistical learning. Hippocampal anticipation based on predictive stimuli correlated with subsequent processing of the predicted stimuli in occipital and parietal cortex, and anticipation in additional brain regions correlated with facilitated object recognition as reflected in behavioral priming. Additional analyses suggested that implicit perceptual anticipation does not contribute to explicit familiarity, but can result in predictive potentiation of category-selective ventral visual cortex. Overall, these findings show that future-oriented processing can arise incidentally during the perception of statistical regularities.


Experimental masticatory muscle pain.
Ten female subjects with no history of temporomandibular disorders performed five exercises designed to induce masticatory muscle pain. Three of the exercises were replications of Christensen's tooth-clenching (1970) and tooth-grinding (1971) studies and Scott and Lundeen's mandibular protrusion (1980) study. The other two exercises were designed to induce specific unilateral masticatory muscle pain. The amount of muscle pain as well as the time of onset and the duration were assessed by a clinical palpation exam and a pain questionnaire. These five pain-inducing methods did not consistently produce masticatory muscle pain in non-pain subjects. However, certain individuals appeared to be very susceptible to developing pain during or after most of the exercises. These susceptible individuals demonstrated a bilateral muscle pain pattern after the unilaterally stressful exercises. None of the five exercises produced a statistically significant amount of site-specific masticatory muscle pain as determined by either the palpation examination or the pain questionnaire, even when the exercise was intended to produce such specificity. The fact that some subjects did develop masticatory muscle pain is indicative that muscle exercise and fatigue may lead to TMD-like symptoms.


Tinnitus in noise-exposed workers.
Tinnitus is said to be a common complaint of workers who are exposed to noise. We studied the prevalence and characteristics of tinnitus in 647 noise-exposed workers who had been notified as cases of noise-induced deafness. One hundred and fifty-one had tinnitus, giving a prevalence of 23.3 per cent. The tinnitus was bilateral in 42.4 per cent of cases, and of high frequency in 44.4 per cent. In 23.8 per cent it was associated with other symptoms. About 30 per cent of those with tinnitus complained that it interfered with daily activities like telephone conversation and sleep. The workers with tinnitus had consistently higher hearing thresholds at both high and low frequencies than those with no tinnitus. This finding remained even after adjusting for differences in sex, age and ethnic group composition and in the noise exposure duration. Workers are often told that noise exposure causes deafness, but little is mentioned about tinnitus. Awareness of the possible occurrence of tinnitus may encourage workers to cooperate more actively in a company hearing conservation programme.


Detection of antibodies in sera from patients with Opisthorchiasis.
The indirect immunofluorescent antibody technique (IFA) was used for detection of antibodies in sera of patients with Opisthorchiasis. Antibodies to fluke worm and egg antigens were detected in 166 of 205 (81%) patients. The test showed that only the IgG class of antibodies reacting exclusively with integumental wall of the worm (AW) were positive in 46.8% (96/205), reacting only with the wall of intact eggs in 11 out of 205 (5.4%) and antibodies to both fluke and their egg antigens were present in 28.8% (59/205). In addition, 5.4% (11/205) of patients' sera were positive for autoantibodies producing a speckled antinuclear antibodies (ANA) pattern. The sera positive for only AW contained detectable autoantibodies to other cell antigens including: anti-smooth muscle antibodies of 9.4% (9/96), antimitochondrial antibodies of 3.1% (3/96), anti-liver/kidney microsomes of 1% (1/96) and anti-parietal cell antibodies of 1% (1/96). Autoantibodies were undetectable in sera from normal subjects. Among the ANA positive sera, 55% (6/11) exhibited antibodies against an extractable nuclear antigen (ENA) by a tanned red cell hemagglutination assay. This finding may suggest that the autoantibody response was due to the cross reaction between worm antigen and self antigen or it may be the result of polyclonal activation of B lymphocytes in these patients.


[Rapid prototyping and bone reconstruction].
The authors describe the main techniques for rapid prototyping used in the medical field, with particular emphasis on Laser stereolithography. They describe the basics of the technique as well as the procedures they currently use to construct anatomic specimens from scanned images. For more than 10 years, the targets of researchers in the field of bone reconstruction in craniofacial surgery have been: to design a system to acquire data directly from CT-scans, to optimize modelling, processing and materials, to verify the possibility of producing prefabricated prostheses with data from preoperative models (22 cases). The target of integrating this technique into therapeutic protocols has been reached. The current trend in research is to produce prostheses from biocompatible materials directly from CT scan data by Laser stereolithography.


Cycloadduct formation of α,β-unsaturated esters with azides catalyzed by NHC systems.
NHC-catalyzed cycloadduct formation of α,β-unsaturated esters with azides has been developed. This strategy could generate 1,2,3-triazoles and dihydropyrazoles with high yields and regioselectivities in the presence of an N-heterocyclic carbene catalyst. The broad substrate scope and mechanistic survey of this process are also presented.


Ocular ischemia and the effects of allopurinol on functional recovery in the retina of the arterially perfused cat eye.
This study sought to examine the acute effects of ocular ischemia and reperfusion on retinal function and determine the extent to which recovery during reperfusion is improved by allopurinol (AP), a blocker of xanthine oxidase (XO). The latter is presumed to be a major factor in the formation of free radicals associated with reperfusion of ischemic tissue. Electroretinographic (ERG) responses were recorded simultaneously from the two isolated, arterially perfused eyes obtained from the same cat. One eye served as the control and received only the standard perfusate; the other eye was infused with AP before and after a 3 hr period of total ischemia. After the prolonged period of nonperfusion, recovery of the electroretinographic components was incomplete to varying degrees. Maximum b-wave amplitudes recovered only to 17 +/- 5% (mean +/- SEM) of baseline, whereas the a-wave grew to 60 +/- 10% of its baseline value. For both measures, the recovery of electroretinographic amplitude was significantly greater in AP-treated eyes than in the control eyes. Generation of free radicals by XO contributes to the retinal damage and loss of function that occurs after a period of ischemia and subsequent reperfusion.


A macromolecular synthesis-dependent late phase of long-term potentiation requiring cAMP in the medial perforant pathway of rat hippocampal slices.
Memory storage consists of a short-term phase that is independent of new protein synthesis and a long-term phase that requires the synthesis of new proteins and RNA. A cellular representation of these two phases has been demonstrated recently for long-term potentiation (LTP) in both the Schaffer collateral and the mossy fibers of the hippocampus, a structure widely thought to contribute to memory consolidation. By contrast, much less information is available about the medial perforant pathway (MPP), one of the major inputs to the hippocampus. We found that both a short-lasting and a long-lasting potentiation (L-LTP) can be induced in the MPP of rat hippocampal slices by applying repeated tetanization in reduced levels of magnesium. This potentiation was dependent on the activation of NMDA receptors. The early, transient phase of LTP in the MPP did not require either protein or RNA synthesis, and it was independent of protein kinase A activation. By contrast, L-LTP required the synthesis of proteins and RNA, and was selectively blocked by inhibitors of cAMP-dependent protein kinase (PKA). Forskolin, an adenylate cyclase activator, also induced a L-LTP that was attenuated by inhibition of transcription. Our results demonstrate that, like LTP in the Schaffer collateral and mossy fiber pathways, MPP LTP also consists of a late phase that is dependent on protein and RNA synthesis and PKA activity. Thus, cAMP-mediated transcription appears to be a common mechanism for the late form of LTP in all three pathways within the hippocampus.


Depressive symptoms and heart failure: examining the sociodemographic variables.
The purpose of this study was to determine the differences in depressive symptoms among a sample of heart failure outpatients by examining sociodemographic and clinical variables: sex, race, marital status, living arrangement/status, heart failure severity, and age. The most frequently reported depressive symptoms were also examined. A descriptive, cross-sectional design was used. Patients were enrolled in a larger research study from 5 clinics in the Midwest (1 adult primary care medicine clinic, 1 heart clinic, and 3 heart failure clinics). The sample included 150 patients with mean age of 61.3 years; 88 (59%) were men, and 62 (41%) were women; 47 (31%) were African American, 101 (67%) were white, and 2 (2%) were Asian patients. Forty-seven percent of the patient sample were New York Heart Association class III. Approximately half (51%) of the patient sample were married. The Patient Health Questionnaire 8 was used to measure depressive symptoms. Heart failure severity was assessed using the New York Heart Association classification. Patients with class III and IV had significantly more depressive symptoms than patients with class I and II (P < .0001). Age was negatively correlated with depressive symptoms scores (P < .0002). There were no significant differences in depressive symptoms among the variables of sex, race, marital status, or living arrangement. The most frequently reported depressive symptom was "feeling tired/no energy." The findings from this study may contribute to the development of a broader knowledge base regarding depressive symptoms and its correlates in heart failure and may be used as a foundation for further research.


PhoP/PhoQ transcriptional repression of Salmonella typhimurium invasion genes: evidence for a role in protein secretion.
Previously, the PhoP-repressed locus prgH was identified as important for signalling epithelial cells to endocytose Salmonella typhimurium. Characterization of prgH revealed that it is an operon of four genes encoding polypeptides of 392 (prgH), 80 (prgI), 101 (prgJ) and 252 amino acid residues (prgK). Synthesis of the 2.6 kb prgHIJK transcript was repressed in bacteria that activate PhoP/PhoQ, indicating that PhoP/PhoQ regulates prgHIJK by transcriptional repression. The prgI, prgJ and prgK predicted gene products were similar to Shigella flexneri and Yersinia enterocolitica proteins required for secretion of Ipa and Yop virulence factors. Analysis of the culture supernatants from wild-type S. typhimurium demonstrated that at least 25 polypeptides larger than 14 kDa could be detected. In contrast, prgH1::TnphoA, phoP-constitutive and hil-deletion mutants had significant defects in their supernatant protein profiles. The invasion and supernatant protein profile defects of the prgH1::TnphoA mutant were both complemented by a 5.1 kb plasmid that included prgHIJK. These results suggest that PhoP/PhoQ regulates extracellular transport of proteins by transcriptional repression of secretion determinants and that secreted proteins may be involved in signalling epithelial cells to endocytose bacteria.


Congenital adrenal tissue in the lung with adrenal cytomegaly. Case report and review of the literature.
Adrenal cytomegaly is a cellular abnormality found in the fetal and neonatal adrenal gland. It is characterized by large polyhedral cells with granular eosinophilic cytoplasm and irregular pleomorphic nuclei. A case of bilateral adrenal cytomegaly in five-day-old infant with two foci of pulmonary involvement is reported with a review of the literature. The question of whether the pulmonary lesion represents metastasis or cytomegalic change in ectopic adrenal tissue is discussed.


A family of bombinin-related peptides from the skin of Bombina variegata.
A peptide fraction was isolated from the skin of Bombina variegata that showed antimicrobial activity. This fraction contained several molecular species, all of them consisting of 27 amino acid residues, with a constant C-terminal region (from residues 14-27), including an amidated carboxyl end and a variable N-terminal segment. These peptides are related but not identical to bombinin [Csordas, A. & Michl, H. (1970) Monatsh. Chem. 101, 182-189]. By using synthetic oligonucleotides corresponding to the C-terminal region of the peptides, a cDNA library from the skin of B. variegata was screened and several positive clones coding for the corresponding peptide precursors were isolated and sequenced. Each clone contained the genetic information for a different bombinin-like peptide. The antimicrobial activity towards different bacterial species of a synthetic peptide corresponding to one of the variants deduced from cDNA sequences was tested. This peptide was found to be mainly active against different isolates of Staphylococci and Escherichia coli.


Genetic structure across urban and agricultural landscapes reveals evidence of resource specialization and philopatry in the Eastern carpenter bee, Xylocopa virginica L.
Human activity continues to impact global ecosystems, often by altering the habitat suitability, persistence, and movement of native species. It is thus critical to examine the population genetic structure of key ecosystemservice providers across human-altered landscapes to provide insight into the forces that limit wildlife persistence and movement across multiple spatial scales. While some studies have documented declines of bee pollinators as a result of human-mediated habitat alteration, others suggest that some bee species may benefit from altered land use due to increased food or nesting resource availability; however, detailed population and dispersal studies have been lacking. We investigated the population genetic structure of the Eastern carpenter bee, Xylocopa virginica, across 14 sites spanning more than 450 km, including dense urban areas and intensive agricultural habitat. X. virginica is a large bee which constructs nests in natural and human-associated wooden substrates, and is hypothesized to disperse broadly across urbanizing areas. Using 10 microsatellite loci, we detected significant genetic isolation by geographic distance and significant isolation by land use, where urban and cultivated landscapes were most conducive to gene flow. This is one of the first population genetic analyses to provide evidence of enhanced insect dispersal in human-altered areas as compared to semi-natural landscapes. We found moderate levels of regional-scale population structure across the study system (G'ST = 0.146) and substantial co-ancestry between the sampling regions, where co-ancestry patterns align with major human transportation corridors, suggesting that human-mediated movement may be influencing regional dispersal processes. Additionally, we found a signature of strong site-level philopatry where our analyses revealed significant levels of high genetic relatedness at very fine scales (<1 km), surprising given X. virginica's large body size. These results provide unique genetic evidence that insects can simultaneously exhibit substantial regional dispersal as well as high local nesting fidelity in landscapes dominated by human activity.


Effect of antioxidants on enzyme-catalysed biodegradation of carbon nanotubes.
The growing applications of carbon nanotubes (CNTs) inevitably increase the risk of exposure to this potentially toxic nanomaterial. In an attempt to address this issue, research has been implemented to study the biodegradation of CNTs. In particular, myeloperoxidase (MPO), an enzyme expressed by inflammatory cells of animals including humans, catalyse the degradation of oxidized carbon nanomaterials. While reactive intermediates generated by MPO efficiently degrade oxidized single-walled carbon nanotubes (o-SWCNTs); the exact mechanism of enzyme-catalysed biodegradation remains ambiguous. In this work, we tried to explain enzymatic oxidation in terms of redox potentials by employing competitive substrates for MPO such as chloride, which is oxidized by MPO to form a strong oxidant (hypochlorite), and antioxidants that have lower redox potentials than CNTs. Employing transmission electron microscopy, Raman spectroscopy, and vis-NIR absorption spectroscopy, we demonstrate that the addition of antioxidants, L-ascorbic acid and L-glutathione, with or without chloride significantly mitigates MPO-catalysed biodegradation of o-SWCNTs. This study focuses on a fundamental understanding of the mechanisms of enzymatic biodegradation of CNTs and the impact of antioxidants on these pathways.


Topological Charge Pumping in Excitonic Insulators.
We show that in excitonic insulators with s-wave electron-hole pairing, an applied electric field (either pulsed or static) can induce a p-wave component to the order parameter, and further drive it to rotate in the s+ip plane, realizing a Thouless charge pump. In one dimension, each cycle of rotation pumps exactly two electrons across the sample. Higher dimensional systems can be viewed as a stack of one-dimensional chains in momentum space in which each chain crossing the Fermi surface contributes a channel of charge pumping. Physics beyond the adiabatic limit, including in particular dissipative effects is discussed.


Transcriptomics and Alternative Splicing Analyses Reveal Large Differences between Maize Lines B73 and Mo17 in Response to Aphid Rhopalosiphum padi Infestation.
Maize (Zea mays L.) is a staple crop worldwide with extensive genetic variations. Various insects attack maize plants causing large yield loss. Here, we investigated the responses of maize B73, a susceptible line, and Mo17, a resistant line, to the aphid Rhopalosiphum padi on metabolite and transcriptome levels. R. padi feeding had no effect on the levels of the defensive metabolites benzoxazinoids (Bxs) in either line, and Mo17 contained substantially greater levels of Bxs than did B73. Profiling of the differentially expressed genes revealed that B73 and Mo17 responded to R. padi infestation specifically, and importantly, these two lines showed large gene expression differences even without R. padi herbivory. Correlation analysis identified four transcription factors (TFs) that might account for the high Bx levels in Mo17. Similarly, genome-wide alternative splicing (AS) analyses indicated that both B73 and Mo17 had temporally specific responses to R. padi infestation, and these two lines also exhibited large differences of AS regulation under normal condition, and 340 genes, including 10 TFs, were constantly differentially spliced. This study provides large-scale resource datasets for further studies on the mechanisms underlying maize-aphid interactions, and highlights the phenotypic divergence in defense against aphids among maize varieties.


Determination of the enthalpy of solute-solvent interaction from the enthalpy of solution: aqueous solutions of erythritol and L-threitol.
In this work the enthalpy of the solute-solvent interaction of erythritol and L-threitol in aqueous solution was determined from the values obtained for the enthalpy of solvation. The values for this property were calculated from those determined for the enthalpies of solution and sublimation. To determine the values of the enthalpy of solute-solvent interaction, the solvation process is considered as taking place in three steps: opening a cavity in the solvent to hold the solute molecule, changing the solute conformation when it passes from the gas phase into solution, and interaction between the solute and the solvent molecules. The cavity enthalpy was calculated by the scaled particle theory and the conformational enthalpy change was estimated from the value of this function in the gas phase and in solution. Both terms were determined by DFT calculations. The solvent effect on the solute conformation in solution was estimated using the CPCM solvation model. The importance of the cavity and conformational terms in the interpretation of the enthalpy of solvation is noted. While the cavity term has been used by some authors, the conformational term is considered for the first time. The structural features in aqueous solution of erythritol and L-threitol are discussed.


[Human cultured lens cells. II. Characterization of established lens cell lines and test of their suitability for cytotoxicity tests].
Human lens cells in culture may be valuable tools to discover cataractogenic risk factors. Here we report on the characterization of established human lens cells and their use in cytotoxicity tests. Adhesion dependence was tested by an autoradiographic method. Soft agar test was performed to secure anchorage-independent growth. Laser scan microscopy was used for measuring the nucleus-cytoplasmic relationship. The cytoskeleton was visualized by fluorescence microscopy. Investigations on cytotoxicity were done by neutral red cytotoxicity assay and 3H-thymidine incorporation. The labeling index was determined by the BrdU-method. Two lens cell lines were transformed into fast growing cells. They are characterized by a partial loss of adhesion dependence, cell growth in soft agar, high cloning efficiency and reduced serum requirement. However, the nucleus-cytoplasmic relationship did not change very much in comparison to non-transformed lens cells. The organization of the cytoskeleton was cell shape dependent. The intermediate filaments were from the vimentin type. The established cell lines proved to be highly sensitive to ethanol and mitomycin C. The established human lens cell lines are well suited to screen for cytotoxic substances in vitro which might be cataractogenic risk factors in vivo.


Anti-N-methyl-D-aspartate receptor and anti-ribosomal-P autoantibodies contribute to cognitive dysfunction in systemic lupus erythematosus.
Autoantibodies against N-methyl-D-aspartate receptor (anti-NMDAR) and ribosomal-P (anti-P) antigens are potential pathogenic factors in the frequently observed diffuse brain dysfunctions in patients with systemic lupus erythematosus (SLE). Although studies have been conducted in this area, the role of anti-NMDAR antibodies in SLE cognitive dysfunction remains elusive. Moreover, the specific contribution of anti-P antibodies has not been reported yet. The present study attempts to clarify the contribution of anti-NMDAR and anti-P antibodies to cognitive dysfunction in SLE. The Cambridge Neuropsychological Test Automated Battery (CANTAB) was used to assess a wide range of cognitive function areas in 133 Chilean women with SLE. ANCOVA models included autoantibodies, patient and disease features. Cognitive deficit was found in 20%. Higher SLEDAI-2K scores were associated with impairment in spatial memory and learning abilities, whereas both anti-NMDAR and anti-P antibodies contributed to deficits in attention and spatial planning abilities, which reflect fronto-parietal cortex dysfunctions. These results reveal an association of active disease together with specific circulating autoantibodies, such as anti-NMDAR and anti-P, with cognitive dysfunction in SLE patients.


Comparative analysis of ear-hole closure identifies epimorphic regeneration as a discrete trait in mammals.
Why mammals have poor regenerative ability has remained a long-standing question in biology. In regenerating vertebrates, injury can induce a process known as epimorphic regeneration to replace damaged structures. Using a 4-mm ear punch assay across multiple mammalian species, here we show that several Acomys spp. (spiny mice) and Oryctolagus cuniculus completely regenerate tissue, whereas other rodents including MRL/MpJ 'healer' mice heal similar injuries by scarring. We demonstrate ear-hole closure is independent of ear size, and closure rate can be modelled with a cubic function. Cellular and genetic analyses reveal that injury induces blastema formation in Acomys cahirinus. Despite cell cycle re-entry in Mus musculus and A. cahirinus, efficient cell cycle progression and proliferation only occurs in spiny mice. Together, our data unite blastema-mediated regeneration in spiny mice with regeneration in other vertebrates such as salamanders, newts and zebrafish, where all healthy adults regenerate in response to injury.


Lack of uniformity in the mutational spectra of chlorohydroxyfuranones in Salmonella typhimurium strain TA100.
The mutational specificity of three chlorohydroxyfuranones found in chlorinated drinking water, 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone (MX), 3-chloro-4(chloromethyl)-5-hydroxy-2(5H)-furanone (CMCF) and 3,4-dichloro-5-hydroxy-2(5H)-furanone (mucochloric acid, MCA), was examined in Salmonella typhimurium strain TA100. DNA colony-hybridization of TA100 revertants showed that MX and CMCF both induced predominantly G:C-->T:A transversions (87 and 75% of total, respectively) with a 3:1 preference for the second position of the hisG46 (CCC) target codon. By contrast, MCA produced primarily G:C-->A:T transitions (66% of the total) with a 4:1 preference for the second position of the CCC codon. The mutational specificity of MCA is consistent with the idea that chloroacetaldehyde, a degradation product of MCA, is responsible for the observed mutations. The chemical mechanism by which either MX or CMCF induces G:C-->T:A transversions remains unknown.


Intra-abdominal cyst following revision of ventriculoperitoneal shunt--case report.
An intra-abdominal cyst is a rare complication of ventriculoperitoneal (VP) shunt. A 19-year-old male was admitted complaining of abdominal pain and distension, dysuria, constipation, headache, and fever. He had undergone a VP shunt for obstructive hydrocephalus caused by a cerebellar astrocytoma 16 years earlier, and had received shunt revision twice, 5 years and 3 months earlier, respectively. Examination on admission revealed neck stiffness, early papilledema, a mass in the lower abdomen, and abdominal muscular guarding with rebound tenderness. Laboratory studies showed leukocytosis of the peripheral blood and pleocytosis of the cerebrospinal fluid (CSF). Abdominal ultrasonograms and computed tomographic scans demonstrated a cystic lesion. Under the diagnosis of meningitis and local peritonitis with an intra-abdominal cyst, we sistemically administered antibiotics and externalized the shunt. However, since the cyst fluid could not be aspirated through the abdominal catheter, it was exchanged with a flexible catheter under fluoroscopic control, according to Seldinger's method. A total of 400 ml of cyst fluid was drained. Staphylococcus epidermidis was detected in both the cyst fluid and the CSF. After meningitis subsided, repositioning of the abdominal catheter into the other side of the abdomen was performed but resulted in shunt malfunction and meningitis due to the same organisms. After meningitis again subsided, the VP shunt was converted to a ventriculoatrial shunt. The clinical course was uneventful thereafter.


CPP mediated insulin delivery: current status and promising future.
A variety of methods including penetrating enhancers, enzyme inhibitors, as well as cargo mediated drug delivery have been explored to improve the intolerance of parenteral administrated insulin, but little success has been achieved so far. Under this background, cell penetrating peptides (CPPs), with their ability to enhance transport efficiency of macromolecular drugs have been demonstrated to be able to increase insulin bioavailability (BA) in a number of studies, of which a BA up to 50.7% relative to subcutaneously administered insulin could be achieved by nasal route under optimal conditions. Furthermore, CPPs could be conveniently formulated with insulin, or be grafted onto drug-loaded cargoes to facilitate the cargo mediated insulin delivery. Here we reviewed the recent achievements on CPP-mediated insulin transport, and outlined various CPP-based delivery strategies which are expected to show potential in clinical translation in the future.


Pyridoxine 5'-phosphate oxidase is correlated with human breast invasive ductal carcinoma development.
Pyridoxine 5'-phosphate oxidase (PNPO) is a converting enzyme for an active form of vitamin B6. This study aims to evaluate the biological function and the regulatory mechanism of PNPO in human breast invasive ductal carcinoma (IDC). We unveiled for the first time that PNPO was upregulated in patients with IDC and was correlated with the overall survival of patients with metastasis at the later stages. Suppression of PNPO inhibited breast cancer cell proliferation, migration, invasion and colony formation, arrested cell cycle at the G2/M phase and induced cell apoptosis. PNPO was positively correlated with lncRNA MALAT1 which was negatively correlated with miR-216b-5p. PNPO was down-regulated and up-regulated by miR-216b-5p mimics and inhibitors, respectively, in breast cancer cells. A microRNA response element was found in both PNPO and MALAT1 transcripts for miR-216b-5p and the dual-luciferase reporter assay confirmed the binding of these transcripts. Knockdown of MALAT1 resulted in an increase of miR-216b-5p and a decrease of PNPO mRNA, indicating a regulatory mechanism of competing endogenous RNAs. Taken together, these results reveal the biological function and a regulatory mechanism of PNPO, in which the MALAT1/miR-216b-5p/PNPO axis may be important in IDC development. Targeting this axis may have therapeutic potential for breast cancer.


Birth number base and its application.
"This paper attempts to present the concept of 'birth number base' as a reflection of the fecundity of women, explain the methods of determining the base, and its application." The birth number base is defined as the number of births in a given year when the total fertility rate is one. The concept is applied to Chinese data from the 1 per 1,000 population survey of 1982 to project the number of births that will have to be prevented in the future if China is to reach its population targets.


Interaction between striatal volume and DAT1 polymorphism predicts working memory development during adolescence.
There is considerable inter-individual variability in the rate at which working memory (WM) develops during childhood and adolescence, but the neural and genetic basis for these differences are poorly understood. Dopamine-related genes, striatal activation and morphology have been associated with increased WM capacity after training. Here we tested the hypothesis that these factors would also explain some of the inter-individual differences in the rate of WM development. We measured WM performance in 487 healthy subjects twice: at age 14 and 19. At age 14 subjects underwent a structural MRI scan, and genotyping of five single nucleotide polymorphisms (SNPs) in or close to the dopamine genes DRD2, DAT-1 and COMT, which have previously been associated with gains in WM after WM training. We then analyzed which biological factors predicted the rate of increase in WM between ages 14 and 19. We found a significant interaction between putamen size and DAT1/SLC6A3 rs40184 polymorphism, such that TC heterozygotes with a larger putamen at age 14 showed greater WM improvement at age 19. The effect of the DAT1 polymorphism on WM development was exerted in interaction with striatal morphology. These results suggest that development of WM partially share neuro-physiological mechanism with training-induced plasticity.


Are all aberrations equal?
To determine for a fixed RMS error (25 microm, over a 6-mm pupil) how each mode of the normalized Zernike polynomial (second through the fourth radial order) affects high and low contrast logMAR visual acuity. Three healthy volunteers served as subjects. CTView was used to generate optically aberrated logMAR charts. Accommodation was paralyzed and pupils dilated. The foveal achromatic axis of the eye was aligned to a 3-mm pupil and the eye was optimally refracted. Aberrated acuity charts were read until five letters were missed. Data were normalized for each subject to the acuity obtained by reading unaberrated charts and plotted as letters lost as a function of Zernike mode. Defocus (Z2(0)) decreased letter acuity more than astigmatism (Z2(2), Z2(-2)). Coma (Z3(1), Z3(-1)) decreased acuity more than trefoil (Z3(3), Z3(-3)). Spherical aberration (Z4(0)) and secondary astigmatism (Z2(2), Z4(-2)) decreased acuity much more than quadrafoil (Z4(4), Z4(-4)). 1. For an equal amount of RMS error not all coefficients of the Zernike polynomial induce equivalent losses in high and low contrast logMAR acuity. 2. Wavefront error concentrated near the center of the pyramid adversely affects visual acuity more than modes near the edge of the pyramid. 3. Large changes in chart appearance are not reflected in equally large decreases in visual performance (ie, subjects could correctly identify highly aberrated letters). 4. Interactions between modes complicate weighting each Zernike mode for visual impact.


An Hsp70 family chaperone, mortalin/mthsp70/PBP74/Grp75: what, when, and where?
Mortalin/mthsp70/PBP74/Grp75 (called mortalin hereafter), a member of the Hsp70 family of chaperones, was shown to have different subcellular localizations in normal and immortal cells. It has been assigned to multiple subcellular sites and implicated in multiple functions ranging from stress response, intracellular trafficking, antigen processing, control of cell proliferation, differentiation, and tumorigenesis. The present article compiles and reviews information on the multiple sites and functions of mortalin in different organisms. The relevance of its differential distributions and functions in normal and immortal cell phenotypes is discussed.


Partner anxiety prior to elective Caesarean section under regional anaesthesia.
This study was designed to measure anxiety levels in 91 partners of women undergoing elective Caesarean section under regional anaesthesia and to outline potential relieving factors. Twenty-eight percent of partners were pathologically anxious. There was a positive association between anxiety and prior attendance at the anaesthetic assessment clinic (p=0.008). There was no statistically significant association between anxiety and education, occupation, gender, relationship to the patient or previous attendance at Caesarean section. Concern about a safe outcome for the mother and child caused most anxiety, less being expressed regarding presence in the operating theatre or anaesthesia per se. Over 70% of partners expressed the view that attendance at the anaesthetic assessment clinic or provision of written information would reduce their anxiety.


cGMP decreases surface NKCC2 levels in the thick ascending limb: role of phosphodiesterase 2 (PDE2).
NaCl absorption in the medullary thick ascending limb of the loop of Henle (THAL) is mediated by the apical Na/K/2Cl cotransporter (NKCC2). Hormones that increase cGMP, such as nitric oxide (NO) and natriuretic peptides, decrease NaCl absorption by the THAL. However, the mechanism by which cGMP decreases NaCl absorption in THALs is not known. We hypothesized that cGMP decreases surface NKCC2 levels in the THAL. We used surface biotinylation to measure surface NKCC2 levels in rat THAL suspensions. We tested the effect of the membrane-permeant cGMP analog dibutyryl-cGMP (db-cGMP) on surface NKCC2 levels. Incubating THALs with db-cGMP for 20 min decreased surface NKCC2 levels in a concentration-dependent manner (basal=100%; db-cGMP 100 microM=77+/-7%; 500 microM=54+/-10% and 1,000 microM=61+/-8%). A different cGMP analog 8-bromo-cGMP (8-Br-cGMP) also decreased surface NKCC2 levels by 25%, (basal=100%; 8-Br-cGMP=75+/-5%). Incubation of isolated, perfused THALs with db-cGMP decreased apical surface NKCC2 labeling levels as measured by immunofluorescence and confocal microscopy. cGMP-stimulated phosphodiesterase 2 (PDE2) mediates the inhibitory effect of NO on NaCl absorption by THALs. Thus we examined the role of PDE2 and found that PDE2 inhibitors blocked the effect of db-cGMP on surface NKCC2. Also, a nonstimulatory concentration of db-cAMP blocked the cGMP-induced decrease in surface NKCC2. Finally, db-cGMP inhibited THAL net Cl absorption by 48+/-4%, and this effect was completely blocked by PDE2 inhibition. We conclude that cGMP decreases NKCC2 levels in the apical membrane of THALs and that this effect is mediated by PDE2. This is an important mechanism by which cGMP inhibits NaCl absorption by the THAL.


Shortcuts in complex engineering systems: a principal-agent approach to risk management.
In this article, we examine the effects of shortcuts in the development of engineered systems through a principal-agent model. We find that occurrences of illicit shortcuts are closely related to the incentive structure and to the level of effort that the agent is willing to expend from the beginning of the project to remain on schedule. Using a probabilistic risk analysis to determine the risks of system failure from these shortcuts, we show how a principal can choose optimal settings (payments, penalties, and inspections) that can deter an agent from cutting corners and maximize the principal's value through increased agent effort. We analyze the problem for an agent with limited liability. We consider first the case where he is risk neutral; we then include the case where he is risk averse.