Why do human diversity levels vary at a megabase scale?
Levels of diversity vary across the human genome. This variation is caused by two forces: differences in mutation rates and the differential impact of natural selection. Pertinent to the question of the relative importance of these two forces is the observation that both diversity within species and interspecies divergence increase with recombination rates. This suggests that mutation and recombination are either directly coupled or linked through some third factor. Here, we test these possibilities using the recently generated sequence of the chimpanzee genome and new estimates of human diversity. We find that measures of GC and CpG content, simple-repeat structures, as well as the distance from the centromeres and the telomeres predict diversity as well as divergence. After controlling for these factors, large-scale recombination rates measured from pedigrees are still significant predictors of human diversity and human-chimpanzee divergence. Furthermore, the correlation between human diversity and recombination remains significant even after controlling for human-chimpanzee divergence. Two plausible and non-mutually exclusive explanations are, first, that natural selection has shaped the patterns of diversity seen in humans and, second, that recombination rates across the genome have changed since humans and chimpanzees shared a common ancestor, so that current recombination rates are a better predictor of diversity than of divergence. Because there are indications that recombination rates may have changed rapidly during human evolution, we favor the latter explanation.


Immunotherapy for Dogs: Still Running Behind Humans.
Despite all good intentions, dogs are still running behind humans in effective cancer immunotherapies. The more effective treatments in humans, like infusions of CAR-T and NK-cells are not broadly pursued for canines due to significant costs, the rather complicated logistics and the lack of targetable surface antigens. Monoclonal antibodies are challenging to develop considering the limited knowledge about canine target antigens and about their mode of action. Although immunogenic vaccines could be less costly, this approach is hampered by the fact that cancer by itself is immuno-suppressive and any preceding chemotherapy may suppress any clinically meaningful immune response. This review - rather than providing a comprehensive listing of all available immunotherapies for dogs, aims at pointing out the issues that are holding back this field but which hopefully can be addressed so that dogs can "catch up" with what is available to humans.


Urinary incontinence in the elderly and in the oldest old: correlation with frailty and mortality.
Urinary incontinence (UI) is very common in the elderly and has personal and social implications. Many authors have pointed out the necessity to analyze UI in correlation with the overall quality of aging, to better understand this syndrome and define measures for its prevention and treatment. In the present study, we addressed this problem by analyzing the UI correlation with frailty, which has emerged in the last decade as the geriatric syndrome correlated with individual homeostatic capacity and then as the basis of the age-related physical decline. In addition, the monitoring of our sample for a long period allowed us to estimate the prognostic significance of UI by analyzing the correlation between UI and mortality. The analysis was performed in a large sample that included numerous ultra-nonagenarians, a population segment that is still poorly known for UI and other geriatric parameters. We found a strict correlation between UI and frailty, suggesting that UI is correlated to the homeostatic and physiological decline leading to frailty. In addition, we found that UI is an independent mortality risk factor in ultra-nonagenarians, suggesting that the neurological sensitivity needed to be continent is lost very soon when the frailty associated physiological decline begins. On the whole, our study suggests that UI is a marker of frailty and that UI patients should be monitored and, in case, treated in a timely manner to avoid, or to limit, the effects of frailty such as malnutrition, falls, and the consequent accumulation of disabilities.


Bacterial Artificial Chromosome Reverse Genetics Approaches for SARS-CoV-2.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a new member of the Coronaviridae family responsible for the coronavirus disease 19 (COVID-19) pandemic. To date, SARS-CoV-2 has been accountable for over 624 million infection cases and more than 6.5 million human deaths. The development and implementation of SARS-CoV-2 reverse genetics approaches have allowed researchers to genetically engineer infectious recombinant (r)SARS-CoV-2 to answer important questions in the biology of SARS-CoV-2 infection. Reverse genetics techniques have also facilitated the generation of rSARS-CoV-2 expressing reporter genes to expedite the identification of compounds with antiviral activity in vivo and in vitro. Likewise, reverse genetics has been used to generate attenuated forms of the virus for their potential implementation as live-attenuated vaccines (LAV) for the prevention of SARS-CoV-2 infection. Here we describe the experimental procedures for the generation of rSARS-CoV-2 using a well-established and robust bacterial artificial chromosome (BAC)-based reverse genetics system. The protocol allows to produce wild-type and mutant rSARS-CoV-2 that can be used to understand the contribution of viral proteins and/or amino acid residues in viral replication and transcription, pathogenesis and transmission, and interaction with cellular host factors.


The Orco gene involved in recognition of host plant volatiles and sex pheromone in the chive maggot Bradysia odoriphaga.
The insect olfactory recognition system plays a crucial role in the feeding and reproductive behaviors of insects. The odorant receptor co-receptor (Orco), as an obligatory chaperone, is critical for odorant recognition by way of forming heteromeric complexes with conventional odorant receptors (ORs). To investigate the biological functions of Orco in perceiving host plant volatiles and sex pheromone, the Orco gene was identified from the chive maggot Bradysia odoriphaga transcriptome data. Multiple sequence alignment reveals that BodoOrco exhibits an extremely high sequence identity with Orcos from other dipteran insects. The expression of BodoOrco is significantly higher in adults than in larvae and pupae, and the BodoOrco gene is primarily expressed in the antennae of both sexes. Furthermore, the Y-tube assay indicated that knockdown of BodoOrco leads to significant reductions in B. odoriphaga adults' response to all tested host plant volatiles. The dsOrco-treated unmated male adults show less attraction to unmated females and responded slowly compared with dsGFP control group. These results indicated that BodoOrco is involved in recognition of sex pheromone and host plant volatiles in B. odoriphaga and has the potential to be used as a target for the design of novel active compounds for developing ecofriendly pest control strategies.


PTPRT epigenetic silencing defines lung cancer with STAT3 activation and can direct STAT3 targeted therapies.
Signal Transducers and Activators of Transcription-3 (STAT3), a potent oncogenic transcription factor, is constitutively activated in lung cancer, but mutations in pathway genes are infrequent. Protein Tyrosine Phosphatase Receptor-T (PTPRT) is an endogenous inhibitor of STAT3 and PTPRT loss-of-function represents one potential mechanism of STAT3 hyperactivation as observed in other malignancies. We determined the role of PTPRT promoter methylation and sensitivity to STAT3 pathway inhibitors in non-small cell lung cancer (NSCLC). TCGA and Pittsburgh lung cancer cohort methylation data revealed hypermethylation of PTPRT associated with diminished mRNA expression in a subset of NSCLC patients. We report frequent hypermethylation of the PTPRT promoter which correlates with transcriptional silencing of PTPRT and increased STAT3 phosphorylation (Y705) as determined by methylation-specific PCR (MSP) and real time quantitative reverse transcription (RT)-PCR in NSCLC cell lines. Silencing of PTPRT using siRNA in H520 lung cancer cell line resulted in increased pSTAT3Tyr705 and upregulation of STAT3 target genes such as Cyclin D1 and Bcl-XL expression. We show this association of PRPRT methylation with upregulation of the STAT3 target genes Cyclin D1 and Bcl-XL in patient derived lung tumour samples. We further demonstrate that PTPRT promoter methylation associated with different levels of pSTAT3Ty705 in lung cancer cell lines had selective sensitivity to STAT3 pathway small molecule inhibitors (SID 864,669 and SID 4,248,543). Our data strongly suggest that silencing of PTPRT by promoter hypermethylation is an important mechanism of STAT3 hyperactivation and targeting STAT3 may be an effective approach for the development of new lung cancer therapeutics.


Developmental exposure of zebrafish to vitamin D receptor acting drugs and environmental toxicants disrupts behavioral function.
Vitamin D receptor (VDR) signaling is important for optimal neurobehavioral development. Disruption of VDR signaling by environmental toxicants during early development might contribute to the etiology of behavioral dysfunction. In the current set of studies, we examined ten compounds known to affect VDR function in vitro for neurobehavioral effects in vivo in zebrafish. Zebrafish embryos were exposed to concentrations of the compounds in their water during the first 5 days post-fertilization. On day 5, the embryos were tested in an alternating light-dark locomotor assay using a computerized video tracking system. We found that most of the compounds produced significant changes in locomotor behavior in exposed zebrafish larvae, although the direction of the effect (i.e., hypo- or hyperactivity) and the sensitivity of the effect to changes in illumination condition varied across the compounds. The nature of the behavioral effects generally corresponded to the effects these compounds have been shown to exert on VDR. These studies lay a foundation for further investigation to determine whether behavioral dysfunction persists into adulthood and if so which behavioral functions are affected. Zebrafish can be useful for screening compounds identified in high throughput in vitro assays to provide an initial test for how those compounds would affect construction and behavioral function of a complex nervous system, helping to bridge the gap between in vitro neurotoxicity assays and mammalian models for risk assessment in humans.


Collagen production and niche engineering: A novel strategy for cancer cells to survive acidosis in DCIS and evolve.
Growing tumors are dynamic and nonlinear ecosystems, wherein cancer cells adapt to their local microenvironment, and these adaptations further modify the environment, inducing more changes. From nascent intraductal neoplasms to disseminated metastatic disease, several levels of evolutionary adaptations and selections occur. Here, we focus on one example of such an adaptation mechanism, namely, "niche construction" promoted by adaptation to acidosis, which is a metabolic adaptation to the early harsh environment in intraductal neoplasms. The avascular characteristics of ductal carcinoma in situ (DCIS) make the periluminal volume profoundly acidic, and cancer cells must adapt to this to survive. Based on discovery proteomics, we hypothesized that a component of acid adaptation involves production of collagen by pre-cancer cells that remodels the extracellular matrix (ECM) and stabilizes cells under acid stress. The proteomic data were surprising as collagen production and deposition are commonly believed to be the responsibility of mesenchymally derived fibroblasts, and not cells of epithelial origin. Subsequent experiments in 3D culture, spinning disk and second harmonic generation microscopy of DCIS lesions in patients' samples are concordant. Collagen production assay by acid-adapted cells in vitro demonstrated that the mechanism of induction involves the RAS and SMAD pathways. Secretome analyses show upregulation of ECM remodeling enzymes such as TGM2 and LOXL2 that are collagen crosslinkers. These data strongly indicate that acidosis in incipient cancers induces collagen production by cancer cells and support the hypothesis that this adaptation initiates a tumor-permissive microenvironment promoting survival and growth of nascent cancers.


Incidence and risk factors for secondary extralaryngeal dystonia in patients with laryngeal dystonia.
Laryngeal dystonia (LD) is a focal dystonia affecting adductor and/or abductor muscles of the larynx. It can be isolated or may spread to extra laryngeal muscles. The aim of this study was to report the characteristics of LD over time in a large single-center study with a long follow-up. Retrospective review of patients with LD referred to our institution between 1991 and 2021. Demographic data, time to diagnosis, type of LD, follow-up and spread of dystonia [SD] were recorded. Risk factors for spread of dystonia during follow-up were analyzed. Over the 30-year period, 516 patients (77.3 % female, median age 50 years, range 5-87 years) were analyzed. Three hundred and fifteen patients (61 %) had adduction laryngeal dystonia, 136 patients (26.4 %) had abduction laryngeal dystonia, 46 patients (8.9 %) had adductor respiratory laryngeal dystonia, 12 patients (2.3 %) had mixed laryngeal dystonia, and seven patients (1.4 %) had singer's laryngeal dystonia. A previous history of dystonia was found in 47 patients (9.1 %). A laryngeal tremor was found in 68 patients (13.2 %). Since the onset of symptoms, LD was diagnosed after a median of 3 years (IQR: 1.0, 7.0). SD occurred in 55 patients (10.7 %) after a median time of 4 year (IQR: 1.5, 13.0). Patients with mixed laryngeal dystonia had higher probability of SD (p = 0.018). This study reports a large European study of LD, with a long follow-up. SD occurred in 10.5 % of patients. Patients with mixed laryngeal dystonia had a higher probability of SD. A close follow-up may be recommended for patients with mixed laryngeal dystonia.


A study on the pre-admission characteristics of patients with acute myocardial infarction.
To clarify an appropriate policy to implement Emergency Medical Services (EMS) in Taiwan based on the profiles of patients with acute myocardial infarction (AMI), and to compare the differences of clinical features accompanied by AMI between Orientals and Westerners, 63 consecutive patients with definite AMI entering the emergency room (ER) from September 1989 through January 1990 were studied. Several interesting facts were found: 1) 9.5% (6/63) of patients took public ambulance to seek medical help; 2) 89% came from out of our jurisdiction, and 62% (39/63) were referred from general practitioners or the ER of community hospitals; 3) There were 5 truly delayed cases (7.9%) owing to a lack of sense of danger in case of chest pain; 4) 50% of our cases occurred during sleeping; 5) The time of onset peaked from 0 AM to 6 AM; 6) While 63.5% of patients visited ER within 6 hours after the onset of acute symptoms, only 12.7% were treated with thrombolytic therapy; and 7) AMI patients stayed in ER for an average of over 4 hours. With limited cases, although trends were evident, firm conclusions were not warranted. The following crucial recommendations are suggested: 1) EMS system in terms of caring of AMI patients still needs a well-designed study to map out an appropriate policy; 2) The strengthening education to physicians working in ER or to all primary care physicians of an early use of thrombolytic agents in indicated cases should be warranted to offer timely therapy when necessary; 3) General public should be educated to call for medical help as soon as possible in case of chest pain.


[Clinical trial of heptral in patients with chronic diffuse liver disease with intrahepatic cholestasis syndrome].
Heptral (S-adenosine-L-methionine) was given to 32 patients with chronic diffuse diseases of the liver and intrahepatic cholestasis. 16 of them had primary biliary cirrhosis (PBC). Phase I of the treatment lasted 16 days when the drug was injected intravenously in a dose 800 mg/day. It was followed by phase 2--1600 mg/day taken for 16 days. A response was registered in the majority of patients. They had relieved symptoms of asthenia, skin pruritus, jaundice. The patients with liver cirrhosis and chronic hepatitis exhibited a statistically significant fall in ALT, AST and GGTP. PBS patients showed insignificant lowering of cholesterol, bilirubin. No resistance was noted in repeated courses. Heptral tolerance was satisfactory.


Facile synthesis of tetrahydroprotoberberine and protoberberine alkaloids from protopines and study on their antibacterial activities.
A series of isoquinoline alkaloids including tetrahydroprotoberberines, N-methyl tetrahydroprotoberberines and protoberberines were facile synthesised with protopines as the starting material. All compounds were evaluated for their antibacterial activities against four pathogenic bacteria Escherichia coli, Staphyloccocus aureus, Staphylococcus gallinarum and Salmonella choleraesuis. Experimental results indicated that protoberberines were the most active compounds to the target bacteria among the tested alkaloids. It was suggested that planar molecule with high aromatisation level (e.g. coptisine 5 and berberine 6) or a positive charge of the molecules (e.g. N-methyl tetrahydroprotoberberines 11 and 12) had a positive influence on the antibacterial effects.


The Stringent Response Induced by Phosphate Limitation Promotes Purine Salvage in Agrobacterium fabrum.
Agrobacterium fabrum induces tumor growth in susceptible plant species. The upregulation of virulence genes that occurs when the bacterium senses plant-derived compounds is enhanced by acidic pH and limiting inorganic phosphate. Nutrient starvation may also trigger the stringent response, and purine salvage is among the pathways expected to be favored under such conditions. We show here that phosphate limitation induces the stringent response, as evidenced by production of (p)ppGpp, and that the xdhCSML operon encoding the purine salvage enzyme xanthine dehydrogenase is upregulated ∼15-fold. The xdhCSML operon is under control of the TetR family transcription factor XdhR; direct binding of ppGpp to XdhR attenuates DNA binding, and the enhanced xdhCSML expression correlates with increased cellular levels of (p)ppGpp. Xanthine dehydrogenase may also divert purines away from salvage pathways to form urate, the ligand for the transcription factor PecS, which in the plant pathogen Dickeya dadantii is a key regulator of virulence gene expression. However, urate levels remain low under conditions that produce increased levels of xdhCSML expression, and neither acidic pH nor limiting phosphate results in induction of genes under control of PecS. Instead, expression of such genes is induced only by externally supplemented urate. Taken together, our data indicate that purine salvage is favored during the stringent response induced by phosphate starvation, suggesting that control of this pathway may constitute a novel approach to modulating virulence. Because bacterial purine catabolism appears to be unaffected, as evidenced by the absence of urate accumulation, we further propose that the PecS regulon is induced by only host-derived urate.


Perspectives on gastrointestinal infections in AIDS.
Gastrointestinal illnesses are among the most common and debilitating complication of infections with HIV, affecting 50 per cent to almost 100 per cent of AIDS patients in developed and developing countries, respectively. A number of factors including relevant modes of transmission, the environment, and immunosuppression conspire to determine which enteric infectious agents HIV-infected persons acquire. In developed countries, transmission of a diverse spectrum of bacteria, viruses, and protozoa is facilitated by unprotected receptive anal intercourse and anal-lingual contact among homosexual men with multiple partners. In developing countries, where most HIV infections occur among heterosexual persons, waterborne and foodborne transmission are the principal modes of transmission of enteric organisms. The severity and duration of symptoms associated with enteric pathogens are determined by the host's immunologic response to the organism. Candida albicans often causes local mucosal disease but less often causes systemic infections in HIV-infected persons, likely because polymorphonuclear cell function is intact. The ability of AIDS patients to control infections with G. lamblia and C. jejuni is related to their ability to mount an antibody response to these organisms during infection. The virulence of the organism may also affect the clinical response to infection. Cryptosporidium causes diarrheal symptoms in both immunocompetent and AIDS patients, but illness is more severe and prolonged in the latter. Giardia lamblia and C. jejuni infections are associated with a range of clinical manifestations in both AIDS patients and HIV-seronegative persons, whereas CMV and possibly adenovirus appear to cause significant disease only among immunocompromised subjects. The availability of effective therapy is among the most decisive factors in determining the duration of enteric infections in AIDS patients. For example, Giardia lamblia may cause acute abdominal pain and diarrhea in HIV-infected subjects but prolonged infections with the parasite are uncommon because effective therapy is available. In contrast, infections with CMV and Cryptosporidium may be severe and chronic as available therapy is generally ineffective or only transiently effective. Awareness of these clinical, epidemiologic, immunologic, and therapeutic aspects of gastrointestinal illness in HIV-infected subjects should help to direct the diagnostic evaluation of these patients and to direct areas of research.


Ocular injuries from automobile batteries.
The incidence of eye injuries related to automobile batteries has sharply increased, currently comprising nearly 1% of all unscheduled eye visits to one medical center. A series of 93 cases obtained over 81/2 years was reviewed and follow-up information obtained. While two thirds of the injuries were relatively minor, 10% (9) of the patients sustained permanent ocular damage or required hospitalization. All of the severe injuries and the majority of the other injuries were caused by battery explosions. Not only should the inherent danger of the lead-acid storage battery be reduced, but the public must be alerted to the hazard.


Extracellular matrix signaling through growth factor receptors during wound healing.
Recently, extracellular matrix components have been shown to contain domains that can interact with and activate receptors with intrinsic tyrosine kinase activity. These receptor tyrosine kinases are strong mediators of the cell responses of proliferation, migration, differentiation, and dedifferentiation. However, an interesting question is raised as to why cells would present growth factor receptor ligands in such a manner, as the majority of growth factors are small, soluble, or only transiently tethered ligands. With the exception of the discoidin domain receptors that bind collagen, the other described domains interact with a receptor that binds ubiquitous soluble peptide growth factors, the epidermal growth factor receptor. Unlike traditional growth factors, these individual "matrikine" domains within tenascin-C, laminin, collagen, and decorin possess relatively low binding affinity (high nanomolar or micromolar) and are often presented in multiple valency. The presentation of ligands within the extracellular matrix in this fashion might allow for unique biochemical and physiological outcomes. This new class of "matrikine" ligand may be critical for wound healing, as the majority of known extracellular matrix components possessing matrikines play a strong role, or are presented uniquely, during skin repair. Tenascin-C expression, for instance, is uniquely regulated spatially and has been proposed to present pro-migratory tracks during skin repair through its epidermal growth factor-like repeats. The epidermal growth factor-like repeats of laminin-5 act as cryptic ligands revealed upon matrix metalloproteinase-2 degradation of the surrounding extracellular matrix. The deletion of the discoidin domain receptors 1 and 2 for collagen have negative consequences on the role of fibroblasts and epithelial cells for matrix metalloproteinase production, migration, proliferation, and extracellular matrix turnover. Finally, decorin can bind to, inhibit, and down-regulate epidermal growth factor receptor levels and signaling, suggesting a tonic role of the epidermal growth factor binding domain of decorin in the resolution of wound healing. We provide a model framework for further studies into this emerging class of signals.


Multiple Antibody Lineages in One Donor Target the Glycan-V3 Supersite of the HIV-1 Envelope Glycoprotein and Display a Preference for Quaternary Binding.
One of the goals of HIV-1 vaccine development is the elicitation of neutralizing antibodies against vulnerable regions on the envelope glycoprotein (Env) viral spike. Broadly neutralizing antibodies targeting the Env glycan-V3 region (also called the N332 glycan supersite) have been described previously, with several single lineages each derived from different individual donors. We used a high-throughput B-cell culture method to isolate neutralizing antibodies from an HIV-1-infected donor with high serum neutralization breadth. Clonal relatives from three distinct antibody lineages were isolated. Each of these antibody lineages displayed modest breadth and potency but shared several characteristics with the well-characterized glycan-V3 antibodies, including dependence on glycans N332 and N301, VH4 family gene utilization, a heavy chain complementarity-determining region 2 (CDRH2) insertion, and a longer-than-average CDRH3. In contrast to previously described glycan-V3 antibodies, these antibodies preferentially recognized the native Env trimer compared to monomeric gp120. These data indicate the diversity of antibody specificities that target the glycan-V3 site. The quaternary binding preference of these antibodies suggests that that their elicitation likely requires the presentation of a native-like trimeric Env immunogen. Broadly neutralizing antibodies targeting the HIV-1 glycan-V3 region with single lineages from individual donors have been described previously. Here we describe three lineages from a single donor, each of which targets glycan-V3. Unlike previously described glycan-V3 antibodies, these mature antibodies bind preferentially to the native Env trimer and weakly to the gp120 monomer. These data extend our knowledge of the immune response recognition of the N332 supersite region and suggest that the mode of epitope recognition is more complex than previously anticipated.


The carcinoembryonic antigen gene family: structure, expression and evolution.
The molecular cloning of carcinoembryonic antigen (CEA) and several cross-reacting antigens reveals a basic domain structure for the whole family, which shows structural similarities to the immunoglobulin superfamily. The CEA family consists of approximately 10 genes which are localized in two clusters on chromosome 19. So far, mRNA species for five of these genes have been identified which show tissue variability in their transcriptional activity. Expression of some of these genes in heterologous systems has been achieved, allowing the localization of some epitopes. The characterization of a CEA gene family in the rat and a comparison with its human counterpart has been utilized in the development of an evolutionary model.


Personalized 177Lu-octreotate peptide receptor radionuclide therapy of neuroendocrine tumours: a simulation study.
Peptide receptor radionuclide therapy (PRRT) with 177Lu-octreotate is commonly administered at empiric, fixed amounts of injected radioactivity (IA). This results in highly variable absorbed doses to critical organs and suboptimal treatment of most patients. The primary aims of this study were to design a personalized PRRT (P-PRRT) protocol based on dosimetry, and to perform a simulation of this protocol in a retrospective cohort of patients with neuroendocrine tumours, in order to assess the potential of P-PRRT to safely increase the absorbed dose to the tumour during a four-cycle induction course. Thirty-six patients underwent 122 fixed-IA 177Lu-octreotate PRRT cycles with quantitative SPECT/CT-based dosimetry. Twenty-two patients completed a four-cycle induction course (29.6 ± 2.4 GBq cumulative IA), with kidney, bone marrow and maximum tumour absorbed doses of 16.2 ± 5.5, 1.3 ± 0.8, and 114 ± 66 Gy, respectively. We simulated a P-PRRT regime in which the renal absorbed dose per IA was predicted by the body surface area and glomerular filtration rate for the first cycle, and by renal dosimetry of the previous cycle(s) for the following cycles. Personalized IA was adjusted at each cycle in order to reach the prescribed renal absorbed dose of 23 Gy over four cycles (with a 25-50% reduction when renal or bone marrow function was impaired). Simulated IA and absorbed doses were based on actual patient characteristics, laboratory values and absorbed doses per IA delivered at each cycle. In the P-PRRT regime, cumulative IA could have been increased to 43.7 ± 16.5 GBq over four induction cycles (10.9 ± 5.0 GBq per cycle), yielding cumulative kidney, bone marrow and maximum tumour absorbed doses of 21.5 ± 2.5, 1.63 ± 0.61, and 163.4 ± 85.9 Gy, respectively. This resulted in an average 1.48-fold increase in cumulative maximum tumour absorbed dose over empiric PRRT (range, 0.68-2.64-fold; P = 0.0013). By standardizing the renal absorbed dose delivered during the induction course, P-PRRT has the potential to significantly increase tumour absorbed dose, thus to augment the therapeutic benefit while limiting toxicity.


Effectiveness of intensive inpatient rehabilitation treatment on disease progression in parkinsonian patients: a randomized controlled trial with 1-year follow-up.
Rehabilitation treatments have acute beneficial effects in Parkinson's disease (PD) patients, but whether the effects persist over time is unclear. To assess whether an intensive rehabilitation treatment (IRT) is effective in improving motor performance compared with a control group in a 12-month follow-up, to investigate whether a second cycle administered after 1 year has the same efficacy as the first treatment, and to determine whether IRT reduces the need for increasing levodopa dosage. A total of 50 PD patients were randomly assigned to 2 groups; 25 participants had 4 weeks of inpatient physical therapy that included treadmill and stabilometric platform training. At discharge, these patients were invited to continue doing the learned exercises. After 12 months, the same treatment was repeated. The control group of 25 patients received only pharmacological treatment and was invited to practice generic physical exercise at home. The rating scales used for the clinical evaluation were the Unified Parkinson's Disease Rating Scale Sections II and III (UPDRS II and III) and total (UPDRS tot). The authors found that the beneficial effects of IRT persisted over time. A second rehabilitation cycle administered after 1 year was as effective as the first treatment. At the end of the study, daily medication dosage was reduced in treated patients, whereas it was significantly increased in control patients. These findings suggest that the natural worsening of symptoms associated with PD can be effectively counteracted by a properly designed IRT.


[The significance of change of Th22 cells in patients with acute lymphoblastic leukemia].
To investigate the proportion of Th22 cells in peripheral blood of patients with acute lymphoblastic leukemia (ALL) and evaluate its significance. The proportions of Th22 cells in peripheral blood of B-ALL and T-ALL patients before therapy (group 1), B-ALL and T-ALL patients in complete remission (ALL-CR, group 2) and healthy donors (group 3) were evaluated by flow cytometry. The cytokines IL-22, TGF-β, TNF-α and IL-6 in peripheral blood of each group were measured by enzyme-linked immunosorbent assay (ELISA). The levels of IL-22 mRNA in peripheral blood mononuclear cells of each group were examined by reverse transcription-PCR (RT-PCR). The percentages of Th22 cells and the levels of IL-22, TNF-α, IL-6 and IL-22 mRNA in B-ALL and T-ALL patients before therapy were (0.44 ± 0.10)%, (10.9 ± 3.4) ng/L, (110.7 ± 26.5) ng/L, (60.2 ± 13.8) ng/L, 0.17 ± 0.04 and (0.46 ± 0.11)%, (11.2 ± 3.5) ng/L, (114.6 ± 27.0) ng/L, (58.7 ± 12.4) ng/L, 0.19 ± 0.04, respectively; Which in B-ALL and T-ALL patients in complete remission were(0.59 ± 0.15)%, (14.3 ± 4.1) ng/L, (142.5 ± 32.7) ng/L, (83.7 ± 18.9) ng/L, 0.25 ± 0.06 and(0.60 ± 0.15)%, (14.6 ± 4.3) ng/L, (140.4 ± 31.4) ng/L, (81.4 ± 18.2) ng/L, 0.26 ± 0.06, significantly lower than those in healthy donors \[(1.24 ± 0.31)%, (19.7 ± 6.6) ng/L, (238.3 ± 50.4) ng/L, (138.0 ± 27.1) ng/L, 0.49 ± 0.09\] (P < 0.01). The percentages of Th22 cells and the levels of IL-22, TNF-α, IL-6 and IL-22 mRNA in group l were lower than those in group 2 (P < 0.05), there was not significant difference between B-ALL and T-ALL (P > 0.05). But the levels of TGF-β in B-ALL and T-ALL patients before therapy \[(30.6 ± 8.2) ng/L, (31.4 ± 8.8) ng/L\] and in complete remission \[(24.2 ± 5.8) ng/L, (25.1 ± 6.1) ng/L\] were significantly higher than those in group 3\[(9.6 ± 2.8) ng/L\] (P < 0.01). However, the level of TGF-β in group 1 was higher than that of group 2 (P < 0.05), there was not significant difference between B-ALL and T-ALL (P > 0.05). Both the number and function of Th22 cells reduced in ALL patients. Th22 cells might be negatively correlated with ALL progression. The lower levels of TNF-α and IL-6, and overexpression of TGF-β in ALL patients might suppress the differentiation of Th22 cells.


The standards of an ultrasound examination of the prostate gland. Part 2.
The paper discusses the rules of the proper performing of the ultrasound examination of the prostate gland. It has been divided into two parts: the general part and the detailed part. The first part presents the necessary requirements related to the ultrasound equipment needed for performing transabdominal and transrectal examinations of the prostate gland. The second part presents the application of the ultrasound examination in benign prostatic hyperplasia, in cases of prostate inflammation and in prostate cancer. Ultrasound examinations applied in the diagnostics of benign prostatic hyperplasia accelerated the diagnosis, facilitated the qualification to surgery and the selection of the treatment method. The assessment of the size of the prostate gland performed using the endorectal ultrasound examination is helpful in making the choice between transurethral electroresection and adenomectomy. In prostate inflammation this examination should be performed with particular gentleness due to pain ailments. The indication for performing the examination in acute inflammation is the suspicion of prostate abscess. In chronic, exacerbating prostatitis it is possible to perform an intraprostatic antibiotic injection. In the recent years increased morbidity and detectability of prostate gland cancer is observed among men. In Poland it ranks second (13%) among diseases occurring in men. The indication for an endorectal examination is the necessity to assess the size of the prostate gland, its configuration, the echostructure in classical ultrasonography, the vascularization in an ultrasound examination performed with power doppler and, if possible, the differences in the gland tissue firmness (consistency) in elastography. The ultrasound examination is used for performing the mapping biopsy of the prostate gland - from routine, strictly defined locations, the targeted biopsy - from locations suspected of neoplastic proliferation and the staging biopsy - from the neurovascular bundles, the seminal vesicles, from the apex of the prostate and from the periprostatic tissue - this type of biopsy is supposed to help in determining local staging of the neoplastic disease. The ultrasound examination is also helpful during the treatment of the neoplasm performed using brachytherapy or using the method of ultrasonic ablation which is still in the phase of clinical trials.


Role of post-ischemic phase-dependent modulation of anti-inflammatory M2-type macrophages against rat brain damage.
Cerebral ischemia triggers inflammatory changes, and early complications and unfavorable outcomes of endovascular thrombectomy for brain occlusion promote the recruitment of various cell types to the ischemic area. Although anti-inflammatory M2-type macrophages are thought to exert protective effects against cerebral ischemia, little has been clarified regarding the significance of post-ischemic phase-dependent modulation of M2-type macrophages. To test our hypothesis that post-ischemic phase-dependent modulation of macrophages represents a potential therapy against ischemic brain damage, the effects on rats of an M2-type macrophage-specific activator, Gc-protein macrophage-activating factor (GcMAF), were compared with vehicle-treated control rats in the acute (day 0-6) or subacute (day 7-13) phase after ischemia induction. Acute-phase GcMAF treatment augmented both anti-inflammatory CD163+ M2-type- and pro-inflammatory CD16+ M1-type macrophages, resulting in no beneficial effects. Conversely, subacute-phase GcMAF injection increased only CD163+ M2-type macrophages accompanied by elevated mRNA levels of arginase-1 and interleukin-4. M2-type macrophages co-localized with CD36+ phagocytic cells led to clearance of the infarct area, which were abrogated by clodronate-liposomes. Expression of survival-related molecules on day 28 at the infarct border was augmented by GcMAF. These data provide new and important insights into the significance of M2-type macrophage-specific activation as post-ischemic phase-dependent therapy.


Eliminating iodine deficiency disorders--the role of the International Council in the global partnership.
Iodine deficiency is the most common preventable cause of brain damage. WHO estimates that some 2.2 billion people are at risk from iodine deficiency in 130 countries. A programme of universal salt iodization was established in 1994 with the aim of eliminating the problem by 2000. This paper reports progress in this field, with particular reference to the primarily scientific role of the International Council for Control of Iodine Deficiency Disorders, a nongovernmental organization founded in 1986. It is now a multidisciplinary network of 600 professionals in 100 countries.


Use of computed tomography in the assessment of mandibular invasion in carcinoma of the retromolar trigone.
Carcinomas originating in the retromolar trigone (RMT) are uncommon and characterized by early spread. Determination of mandibular invasion is significant for planning therapy and determining prognosis. For oral cavity cancers in general, CT is reasonably accurate in assessing bone invasion. However, there is a paucity of information specifically addressing the value of CT in the RMT. In this study, the records of patients with biopsy-proven RMT carcinomas treated between 1984 and 1998 were reviewed with attention to preoperative CT scans and histopathologic findings during surgery. Half of the patients who were treated with primary resection had mandibular invasion. Bone invasion was not identified radiographically in 27% of patients with preoperative CT scans. The sensitivity of CT for bone involvement in RMT cancers was 50%, with a negative predictive value of 61.1%. The positive predictive value was 91.1%. These findings suggest that CT is a useful, but potentially inaccurate, predictor of bone invasion in the RMT.


Tobacco use among high school athletes and nonatieletes: results of the 1997 youth risk behavior survey.
In the United States today, the use of tobacco has become an entrenched part of teenage culture. The present study used the 1997 Youth Risk Behavior Survey (YRBS), which collected data from a nationally representative sample of 16,262 students in public and private high schools, to compare the tobacco use patterns of athletes and nonathletes. The independent variable, athletic participation, differentiated between moderately involved (1 or 2 teams) and highly involved (3 or more teams) athletes. Frequency of cigarette and cigar smoking and smokeless tobacco use served as the operational measure of tobacco use. Age, race/ethnicity, parental education, and residence were controlled. Logistic regression was used to calculate odds ratios for female and male athletes and nonathletes for each of the tobacco use variables. It was found that both male and female athletes were less likely to have ever smoked regularly, the effect being stronger for more highly involved athletes of both genders. Cigar smoking was unrelated to athlete status. Both female and male athletes were more likely to have used smokeless tobacco, the effect being stronger for more highly involved athletes of both genders. The findings are discussed in terms of access to health information, performance considerations, social status factors, the salience of an athletic identity, and the influence of the athletic subculture on its members.


[Electron microscopic-cytochemical study of succinate dehydrogenase activity of nerve tissue cells of human embryos in vitro].
The ultrastructural localization of succinate dehydrogenase (SDH) activity was revealed in astrocyte-like cells of 26 days old nervous tissue organotypic cultures. The Kerpel-Fronius and Hajos (1968) technique was employed using potassium ferricyanide as an artificial acceptor of electrons. The localization of the reaction product was seen more often in cells with long, thread-like mitochondria. The appearance of such organelles with a large membrane length and high enzymatic activity is suggested to reflect one of the mechanisms of adaptation of nervous tissue cells providing the adequate level of the energy supply under in vitro conditions.


Complementary and alternative medicine use in patients with haematological malignancies in Europe.
This study reports upon a descriptive cross-sectional survey assessing the use of complementary and alternative medicine (CAM) in patients with haematological cancers. Twelve European countries contributed data from patients with haematological cancers, as part of a larger study. Sixty-eight patients with haematological cancer participated. Among the participants, 26.5% used some form of CAM after the cancer diagnosis. The most common therapies used were homeopathy (38.9%), herbal medicine (22.2%) various psychic therapies, such as use of mediums, healers, rebirthing or past life regression therapy (22.2%). A particular profile of a CAM user was not evident in the sample. Moderate levels of satisfaction with CAM were reported. Patients commonly used CAM to increase the ability of their body to fight cancer and to improve physical and emotional well-being. Information about CAM was received mainly from friends or family. As CAM use in patients with haematological malignancies is common, clinicians should assist patients who want to use CAM to make an appropriate decision, and improve communication with them about CAM use in an open and non-judgemental dialogue.


Parental knowledge, beliefs and behaviours for oral health of toddlers residing in rural Victoria.
Little is known about the oral health of children under the age of four years. The determinants of early childhood caries (ECC) in this young age group are also not well understood despite a growing recognition that early interventions may deliver the greatest benefits. The aim of this study was to examine the oral health-related knowledge, attitudes and reported behaviours of parents of children aged 12-24 months living in rural areas of Victoria, Australia. A robust theoretical model was utilized to identify oral health-related behaviours and their antecedent and reinforcing conditions within the context of this specific population group. Two hundred and ninety-four parent/child dyads were recruited through their maternal and child health nurses as part of a larger intervention trial. Parents completed a self-report questionnaire. Knowledge regarding risk and protective factors amongst parents was variable and sometimes at odds with contemporary evidence. Knowledge of the role of early infection with S. mutans was very low, with high levels of behaviours that may promote early transmission reported. Tooth cleaning was reported by most parents at least sometimes, however a large proportion lacked confidence and this was significantly related to the frequency of the cleaning. Parents were confused about the fluoride status of their water supplies. Most parents believed fluoride toothpaste reduced the risk of ECC but did not know whether it should be used with toddlers. The results of this study have implications for efforts to prevent dental decay in this very young age group. Health care professionals other than dentists need support to provide information and promote confidence with regard to optimal fluoride exposure. Attention should also be given to the contribution of early contact with particular bacteria in oral health education and promotion programmes.


What's in a game: the effect of social motivation on interference control in boys with ADHD and autism spectrum disorders.
Children with attention deficit hyperactivity disorder (ADHD) and with autism spectrum disorders (ASD) are known to have cognitive control deficits. Some studies suggest that such deficits may be reduced when motivation is increased through tangible reinforcers. Whether these deficits can also be modulated by non-tangible reinforcers has hardly been studied. Therefore, the effect of social motivation on the ability to suppress irrelevant information (i.e., interference control) was investigated in 22 ADHD boys, 22 ASD boys, and 33 typically developing (TD) boys. An adapted Eriksen Flanker task was administered under a motivational condition in which the boys were told that they were competing with peers, and under a neutral condition in which standard instructions were given. In comparison with TD boys, boys with ADHD were impaired even when no interference was present, while this was not the case for the ASD boys. All groups benefited from the motivation manipulation, i.e., their performance increased when they thought they were competing with peers. Although the boys with ADHD were still slower than TD boys when motivated, they performed as accurately as TD boys. Children with ASD also improved slightly in accuracy and response speed, but this did not reach significance. Children with ADHD are able to exert sufficient cognitive control when they are motivated, which is in line with the current models of ADHD. However, motivation seems to have a general effect on performance and is not solely related to cognitive control abilities. In contrast, this effect was not obtained in children with ASD.


Lenalidomide versus bortezomib maintenance after frontline autologous stem cell transplantation for multiple myeloma.
Lenalidomide (LEN) maintenance (MT) post autologous stem cell transplantation (ASCT) is standard of care in newly diagnosed multiple myeloma (MM) but has not been compared to other agents in clinical trials. We retrospectively compared bortezomib (BTZ; n = 138) or LEN (n = 183) MT from two subsequent GMMG phase III trials. All patients received three cycles of BTZ-based triplet induction and post-ASCT MT. BTZ MT (1.3 mg/m2 i.v.) was administered every 2 weeks for 2 years. LEN MT included two consolidation cycles (25 mg p.o., days 1-21 of 28 day cycles) followed by 10-15 mg/day for 2 years. The BTZ cohort more frequently received tandem ASCT (91% vs. 33%) due to different tandem ASCT strategies. In the LEN and BTZ cohort, 43% and 46% of patients completed 2 years of MT as intended (p = 0.57). Progression-free survival (PFS; HR = 0.83, p = 0.18) and overall survival (OS; HR = 0.70, p = 0.15) did not differ significantly with LEN vs. BTZ MT. Patients with <nCR after first ASCT were assigned tandem ASCT in both trials. In patients with <nCR and tandem ASCT (LEN: n = 54 vs. BTZ: n = 84), LEN MT significantly improved PFS (HR = 0.61, p = 0.04) but not OS (HR = 0.46, p = 0.09). In conclusion, the significant PFS benefit after eliminating the impact of different tandem ASCT rates supports the current standard of LEN MT after ASCT.


Design, Microstructure and Mechanical Properties of Cast Medium Entropy Aluminium Alloys.
In this work, the design, microstructures and mechanical properties of five novel non-equiatomic lightweight medium entropy alloys were studied. The manufactured alloys were based on the Al65Cu5Mg5Si15Zn5X5 and Al70Cu5Mg5Si10Zn5X5 systems. The formation and presence of phases and microstructures were studied by introducing Fe, Ni, Cr, Mn and Zr. The feasibility of CALPHAD method for the design of new alloys was studied, demonstrating to be a good approach in the design of medium entropy alloys, due to accurate prediction of the phases, which were validated via X-ray diffraction and scanning electron microscopy with energy dispersive spectroscopy. In addition, the alloys were manufactured using an industrial-scale die-casting process to make the alloys viable as engineering materials. In terms of mechanical properties, the alloys exhibited moderate plastic deformation and very high compressive strength up to 644 MPa. Finally, the reported microhardness value was in the range of 200 HV0.1 to 264 HV0.1, which was two to three times higher than those of commercial Al alloys.


The development of vestibulocochlear efferents and cochlear afferents in mice.
We have reinvestigated the embryonic development of the vestibulocochlear system in mice using anterograde and retrograde tracing techniques. Our studies reveal that rhombomeres 4 and 5 include five motor neuron populations. One of these, the abducens nucleus, will not be dealt with here. Rhombomere 4 gives rise to three of the remaining populations: the facial branchial motor neurons; the vestibular efferents; and the cochlear efferents. The migration of the facial branchial motor neurons away from the otic efferents is completed by 13.5 days post coitum (dpc). Subsequently the otic efferents separate into the vestibular and cochlear efferents, and complete their migration by 14.5 dpc. In addition to their common origin, all three populations have perikarya that migrate via translocation through secondary processes, form a continuous column upon completion of their migrations, and form axonal tracts that run in the internal facial genu. Some otic efferent axons travel with the facial branchial motor nerve from the internal facial genu and exit the brain with that nerve. These data suggest that facial branchial motor neurons and otic efferents are derived from a common precursor population and use similar cues for pathway recognition within the brain. In contrast, rhombomere 5 gives rise to the fourth population to be considered here, the superior salivatory nucleus, a visceral motor neuron group. Other differences between this group and those derived from rhombomere 4 include perikaryal migration as a result of translocation first through primary processes and only then through secondary processes, a final location lateral to the branchial motor/otic efferent column, and axonal tracts that are completely segregated from those of the facial branchial and otic efferents throughout their course inside the brain. Analysis of the peripheral distribution of the cochlear efferents and afferents show that efferents reach the spiral ganglion at 12.5 dpc when postmitotic ganglion cells are migrating away from the cochlear anlage. The efferents begin to form the intraganglionic spiral bundle by 14.5 dpc and the inner spiral bundle by 16.5 dpc in the basal turn. They have extensive collaterals among supporting cells of the greater epithelial ridge from 16.5 dpc onwards. Afferents and efferents in the basal turn of the cochlea extend through all three rows of outer hair cells by 18.5 dpc. Selective labeling of afferent fibers at 20.5 dpc (postnatal day 1) shows that although some afferents are still in early developmental stages, some type II spiral ganglion cells already extend for long distances along the outer hair cells, and some type I spiral ganglion cells end on a single inner hair cell. These data support previous evidence that in mice the early outgrowth of afferent and efferent fibers is essentially achieved by birth.


Toward optimal tissue sealants for neurosurgery: use of a novel hydrogel sealant in a canine durotomy repair model.
Watertight dural repairs are difficult to achieve, and cerebrospinal fluid leakage causes complications and extends hospital stays. Therefore, a novel synthetic hydrogel film was evaluated as an adjunct to dural closure in a canine model. The self-polymerizing, absorbable, and biocompatible hydrogel was sprayed onto tissue and formed a flexible, adherent sealant. A 2-cm incision of cranial dura and arachnoid was created in 26 adult dogs and loosely repaired. Hydrogel was applied over the 2-mm dural gap in 13 dogs; 13 control dogs received no hydrogel application. All dogs remained neurologically intact. Valsalva tests conducted at 1, 4, 7, and 56 days were associated with mean leakage pressures (+/- standard error of the mean) of 5 +/- 0, 5 +/- 0, 7 +/- 2, and 13 +/- 8 cm H(2)O in the controls and of 53 +/- 2, 37 +/- 11, 42 +/- 6, and 48 +/- 4 in the treated animals (P = 0.001, 0.053, 0.010, 0.035, respectively, at each time point; one-tailed t test). Histopathological analysis revealed minimal changes. The hydrogel-treated animals exhibited normal progression of dural healing, no dural adhesions, and no underlying effects on the brain. Although dural healing progressed normally, the control animals displayed marked peridural adhesions. The results of this in vivo study suggest that hydrogels, such as that used here, may significantly decrease cerebrospinal fluid leakage, thereby increasing the safety and effectiveness of dural closure in patients and facilitating surgical reexploration.


Motorcycle taxi programme increases safe riding behaviours among its drivers in Kampala, Uganda.
SafeBoda is a motorcycle taxi company that provides road safety training and helmets to its drivers in Kampala, Uganda. We sought to determine whether SafeBoda drivers are more likely to engage in safe riding behaviours than regular drivers (motorcycle taxi drivers not part of SafeBoda). METHODS : We measured riding behaviours in SafeBoda and regular drivers through: (1) computer-assisted personal interview (CAPI), where 400 drivers were asked about their riding behaviours (eg, helmet and mobile phone use) and (2) roadside observation, where riding behaviours were observed in 3000 boda-boda drivers and their passengers along major roads in Kampala. Across the two cross-sectional studies, a higher proportion of SafeBoda drivers than regular drivers engaged in safe riding behaviours. For instance, helmet use among SafeBoda compared with regular drivers was 21% points higher (95% CI 0.15 to 0.27; p<0.001) based on the CAPI and 45% points higher (95% CI 0.43 to 0.47; p<0.001) based on roadside observation. Furthermore, compared with regular drivers, SafeBoda drivers were more likely to report having a driver's license (66.3% vs 33.5 %; p<0.001) and a reflective jacket (99.5% vs 50.5 %; p<0.001) and were less likely to report driving towards oncoming traffic (4% vs 45.7 %; p<0.001) in the past 30 days. The SafeBoda programme is associated with increased safe riding behaviours among motorcycle taxi drivers in Kampala. Therefore, the promotion and expansion of such programmes may lead to a reduction in morbidity and mortality due to road injuries.


KLF5 promotes hypoxia-induced survival and inhibits apoptosis in non-small cell lung cancer cells via HIF-1α.
Transcription factor Krüppel-like factors 5 (KLF5) is overexpressed in a wide range of tumor tissues and acts as a prognostic factor in cancer. However, the role of KLF5 in non-small cell lung cancer is not clear. Hypoxia plays a vital part in the development of cancer via hypoxia-inducible factor 1 (HIF-1). Our study showed that hypoxia (1% O2) increased cell viability, clonality and proliferation and inhibited cell apoptosis in A549 cells. The expression of HIF-1α and KLF5 was increased time-dependently in hypoxia. Using small interfering RNA (siRNA) targeting KLF5 or HIF-1α, we demonstrated that KLF5 or HIF-1α knockdown inhibited hypoxia-induced cell survival and promoted cell apoptosis by actively downregulating cyclin B1, survivin and upregulating caspase-3. Given the similar effect of KLF5 and HIF-1α on cell survival, an attempt was made to investigate the putative interaction of them in hypoxia. KLF5 was revealed to co-immunoprecipitate with HIF-1α and hypoxia increased the amount of KLF5 and HIF-1α complex. Moreover, silencing of KLF5 decreased HIF-1α expression while KLF5 was not affected by HIF-1α inhibition in hypoxia, confirming the effect of KLF5 on upregulation of HIF-1α. In conclusion, this study identified hypoxia as a tumor promoter by triggering KLF5 → HIF-1α → cyclin B1/survivin/caspase-3 in lung cancer cells.


Berberine-Loaded Carboxylmethyl Chitosan Nanoparticles Ameliorate DSS-Induced Colitis and Remodel Gut Microbiota in Mice.
Inflammatory bowel disease (IBD) is a refractory disorder characterized by chronic and recurrent inflammation. The progression and pathogenesis of IBD is closely related to oxidative stress and irregularly high concentrations of reactive oxygen species (ROS). A new oxidation-responsive nano prodrug was constructed from a phenylboronic esters-modified carboxylmethyl chitosan (OC-B) conjugated with berberine (BBR) that degrades selectively in response to ROS. The optimized micelles exhibited well-controlled physiochemical properties and stability in a physiological environment. OC-B-BBR micelles could effectively encapsulate the anti-inflammatory drug berberine and exhibit ideal H2O2-triggered release behavior as confirmed by in vitro drug loading and release studies. The in vivo anti-inflammatory effect and regulation of gut microbiota caused by it were explored in mice with colitis induced by dextran sodium sulfate (DSS). The results showed that OC-B-BBR significantly ameliorated colitis symptoms and colon damage by regulating the expression levels of IL-6 and remodeling gut microbiota. In summary, this study exhibited a novel BBR-loaded Carboxylmethyl Chitosan nano delivery system which may represent a promising approach for improving IBD treatment.


Crystal structure of a Rad51 filament.
Rad51, the major eukaryotic homologous recombinase, is important for the repair of DNA damage and the maintenance of genomic diversity and stability. The active form of this DNA-dependent ATPase is a helical filament within which the search for homology and strand exchange occurs. Here we present the crystal structure of a Saccharomyces cerevisiae Rad51 filament formed by a gain-of-function mutant. This filament has a longer pitch than that seen in crystals of Rad51's prokaryotic homolog RecA, and places the ATPase site directly at a new interface between protomers. Although the filament exhibits approximate six-fold symmetry, alternate protein-protein interfaces are slightly different, implying that the functional unit of Rad51 within the filament may be a dimer. Additionally, we show that mutation of His352, which lies at this new interface, markedly disrupts DNA binding.


Distribution of ferritin, transferrin and lactoferrin in breast carcinoma tissue.
An immunoperoxidase staining technique was used for detecting three major iron binding proteins (ferritin, transferrin and lactoferrin) in 40 breast carcinoma cases and six benign breast proliferative lesions. Ferritin staining was detected mainly in connectival stroma and in histiocytes surrounding neoplastic cells. Few and faint ferritin positivities were also detected in neoplastic cells of 20 carcinoma cases. Transferrin was found inconsistently in myoepithelial cells surrounding normal ductules, or around neoplastic ducts of ductal in situ carcinoma. In eight carcinoma cases, transferrin staining was also positive in neoplastic cells. Lactoferrin was detected only in normal breast epithelial cells and in benign breast proliferative lesions. These immunohistochemical findings may suggest that raised serum ferritin concentrations in breast carcinoma patients might be attributed to stromal reaction rather than to tumour synthesis. Transferrin staining of neoplastic cells in these carcinoma cases appears to be very intriguing, particularly since transferrin is considered an obligate requirement for growing cells, and transferrin receptors have been demonstrated only in dividing cells. On the basis of the immunohistochemical data, lactoferrin might be used as a pointer to benign lesions.


Functional indicators of response mechanisms to nitrogen deposition, ozone, and their interaction in two Mediterranean tree species.
The effects of nitrogen (N) deposition, tropospheric ozone (O3) and their interaction were investigated in two Mediterranean tree species, Fraxinus ornus L. (deciduous) and Quercus ilex L. (evergreen), having different leaf habits and resource use strategies. An experiment was conducted under controlled condition to analyse how nitrogen deposition affects the ecophysiological and biochemical traits, and to explore how the nitrogen-induced changes influence the response to O3. For both factors we selected realistic exposures (20 kg N ha-1 yr-1 and 80 ppb h for nitrogen and O3, respectively), in order to elucidate the mechanisms implemented by the plants. Nitrogen addition resulted in higher nitrogen concentration at the leaf level in F. ornus, whereas a slight increase was detected in Q. ilex. Nitrogen enhanced the maximum rate of assimilation and ribulose 1,5-bisphosphate regeneration in both species, whereas it influenced the light harvesting complex only in the deciduous F. ornus that was also affected by O3 (reduced assimilation rate and accelerated senescence-related processes). Conversely, Q. ilex developed an avoidance mechanism to cope with O3, confirming a substantial O3 tolerance of this species. Nitrogen seemed to ameliorate the harmful effects of O3 in F. ornus: the hypothesized mechanism of action involved the production of nitrogen oxide as the first antioxidant barrier, followed by enzymatic antioxidant response. In Q. ilex, the interaction was not detected on gas exchange and photosystem functionality; however, in this species, nitrogen might stimulate an alternative antioxidant response such as the emission of volatile organic compounds. Antioxidant enzyme activity was lower in plants treated with both O3 and nitrogen even though reactive oxygen species production did not differ between the treatments.


Inhibition of mitogen-activated protein kinase by a Drosophila dual-specific phosphatase.
The Drosophila extracellular signal-regulated kinase (DERK) mitogen-activated protein kinase (MAPK) is involved in the regulation of multiple differentiation and developmental processes. Tight control of MAPK activity is critical for normal cell behaviour. We identified a novel Drosophila MAPK phosphatase (DMKP) cDNA from the expressed-sequence-tag database and characterized it. Analysis of the nucleotide sequence revealed an open reading frame encoding the 203-amino acid protein, with a calculated molecular mass of 23 kDa, which has a high amino acid sequence similarity with 'VH1-like' dual-specific phosphatases at the broad region near the catalytic sites. The expression of DMKP mRNA occurs from the late larval stages to adulthood in Drosophila development. The recombinant DMKP protein produced in yeast retained its phosphatase activity. When expressed in Schneider cells, DMKP dose-dependently inhibited DERK and Drosophila c-Jun N-terminal kinase activities with high selectivity towards DERK. However, DMKP did not have any affect on Drosophila p38 activity. When DMKP was expressed in yeast, it down-regulated the fus1-lacZ trans-reporter gene of the pheromone MAPK pathway without any significant effect on the high-osmolarity-glycerol-response pathway.


Translation and Validation of Chronic Otitis Media Benefit Inventory (COMBI) in Turkish Language.
The aim of this study was to translate the Chronic Otitis Media Benefit Inventory (COMBI) into Turkish and evaluate the questionnaire's reliability in assessing benefit outcomes in patients after ear surgery. A total of 30 patients with chronic otitis media who underwent ear surgery were asked to fill out the translated Turkish version of the COMBI questionnaire six months after their surgery twice, 10 days apart. The patients were divided into two groups in two different ways based on surgical and audiological outcomes. These were surgical success (SS), surgical failure (SF), audiological success (AS), audiological failure (AF) groups. Student's -t test was performed to show the differences in the COMBI scores of the groups. Cronbach's alpha value was calculated to test internal consistency, Pearson's correlation coefficient was measured to demonstrate test-retest reliability and factor analysis was run. The average score of the SS group was 51.08 and significantly higher than the score of the SF group (42.40) (p=0.012). There was, however, no statistically significant difference between the average scores of the AS and the AF groups (p=0.330). A Cronbach's alpha value of 0.862 was found. Pearson's r value was calculated 0.810. Factor analysis showed that a three-factor solution explained the 70.2% of the variance. The Turkish version of the COMBI is a valid test with high test-retest reliability. It should be used with objective assessment tools for better understanding how patients benefit from the surgery.


N-(4-Hydroxyphenyl)retinamide increases dihydroceramide and synergizes with dimethylsphingosine to enhance cancer cell killing.
Fenretinide [N-(4-hydroxyphenyl)retinamide (4-HPR)] is cytotoxic in many cancer cell types. Studies have shown that elevation of ceramide species plays a role in 4-HPR cytotoxicity. To determine 4-HPR activity in a multidrug-resistant cancer cell line as well as to study ceramide metabolism, MCF-7/AdrR cells (redesignated NCI/ADR-RES) were treated with 4-HPR and sphingolipids were analyzed. TLC analysis of cells radiolabeled with [3H]palmitic acid showed that 4-HPR elicited a dose-responsive increase in radioactivity migrating in the ceramide region of the chromatogram and a decrease in cell viability. Results from liquid chromatography/electrospray tandem mass spectrometry revealed large elevations in dihydroceramides (N-acylsphinganines), but not desaturated ceramides, and large increases in complex dihydrosphingolipids (dihydrosphingomyelins, monohexosyldihydroceramides), sphinganine, and sphinganine 1-phosphate. To test the hypothesis that elevation of sphinganine participates in the cytotoxicity of 4-HPR, cells were treated with the sphingosine kinase inhibitor d-erythro-N,N-dimethylsphingosine (DMS), with and without 4-HPR. After 24 h, the 4-HPR/DMS combination caused a 9-fold increase in sphinganine that was sustained through +48 hours, decreased sphinganine 1-phosphate, and increased cytotoxicity. Increased dihydrosphingolipids and sphinganine were also found in HL-60 leukemia cells and HT-29 colon cancer cells treated with 4-HPR. The 4-HPR/DMS combination elicited increased apoptosis in all three cell lines. We propose that a mechanism of 4-HPR-induced cytotoxicity involves increases in dihydrosphingolipids, and that the synergy between 4-HPR and DMS is associated with large increases in cellular sphinganine. These studies suggest that enhanced clinical efficacy of 4-HPR may be realized through regimens containing agents that modulate sphingoid base metabolism.


Large eddy simulation of the gas-particle turbulent wake flow.
To find out the detailed characteristics of the coherent structures and associated particle dispersion in free shear flow, large eddy simulation method was adopted to investigate a two-dimensional particle-laden wake flow. The well-known Sub-grid Scale mode introduced by Smagorinsky was employed to simulate the gas flow field and Lagrangian approach was used to trace the particles. The results showed that the typical large-scale vortex structures exhibit a stable counter rotating arrangement of opposite sign, and alternately form from the near wall region, shed and move towards the downstream positions of the wake with the development of the flow. For particle dispersion, the Stokes number of particles is a key parameter. At the Stokes numbers of 1.4 and 3.8 the particles concentrate highly in the outer boundary regions. While the particles congregate densely in the vortex core regions at the Stokes number of 0.15, and the particles at Stokes number of 15 assemble in the vortex braid regions and the rib regions between the adjoining vortex structures.


Effects of endogenous sulfur dioxide on monocrotaline-induced pulmonary hypertension in rats.
The present study aimed to explore the protective effect of endogenous sulfur dioxide (SO2) in the development of monocrotaline (MCT)-induced pulmonary hypertension (PH) in rats. Forty Wistar rats were randomly divided into the MCT group receiving MCT treatment, the MCT+L-aspartate-beta- hydroxamate (HDX) group receiving MCT plus HDX treatment, the MCT+SO2 group receiving MCT plus SO2 donor treatment, and the control group. Mean pulmonary artery pressure (mPAP) and structural changes in pulmonary arteries were evaluated. SO2 content, aspartate aminotransferase activity, and gene expression were measured. Superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT), reduced glutathione (GSH), oxidized glutathione, and malondialdehyde (MDA) levels were assayed. In the MCT-treated rats, mPAP and right ventricle/(left ventricle+septum) increased significantly (P<0.01), pulmonary vascular structural remodeling developed, and SOD, GSHPx, CAT, GSH, and MDA levels of lung homogenates significantly increased (P<0.01) in association with the elevated SO2 content, aspartate aminotransferase activity, and gene expression, compared with the control rats. In the MCT+HDXtreated rats, lung tissues and plasma SO2 content and aspartate aminotransferase activities decreased significantly, whereas the mPAP and pulmonary vascular structural remodeling were markedly aggravated with the decreased SOD, CAT, and GSH levels of lung tissue homogenates compared with the MCT-treated rats (P<0.01). In contrast, with the use of a SO2 donor, the pulmonary vascular structural remodeling was obviously lessened with elevated lung tissue SOD, GSH-Px, and MDA content, and plasma SOD, GSH-Px, and CAT levels. Endogenous SO2 might play a protective role in the pathogenesis of MCT-induced PH and promote endogenous antioxidative capacities.


The prognostic role of c-MYC amplification in schistosomiasis-associated colorectal cancer.
The purpose of this study was to explore the prognostic role of c-MYC amplification in colorectal cancer, particularly in schistosomiasis-associated colorectal cancer. Three hundred and fifty four cases of colorectal cancer, which were from Qingpu Branch of Zhongshan Hospital affiliated to Fudan University, were retrospectively analyzed in a tissue microarray (TMA) format, with fluorescence in situ hybridization (FISH) assay and immunohistochemistry (IHC). c-MYC gene amplification was found in 14.1% (50 out of 354) of patients with colorectal cancer and was correlated with old age (P = 0.028), positive lymph node metastasis (P = 0.004) and advanced stage tumors (P = 0.002). The overexpression of c-MYC was closely associated with the amplification status (P = 0.023). Kaplan-Meier survival curves for overall survival (OS) showed a statistically significant difference for patients with c-MYC amplification in full cohort of colorectal cancer, stage III-IV set and patients with lymph node metastasis (P = 0.002, 0.034, 0.012, respectively). Further analysis found c-MYC amplification associated with poorer survival in the subgroup of colorectal cancer with schistosomiasis (CRC-S, P < 0.001), but not in colorectal cancer without schistosomiasis (CRC-NS, P = 0.155). By multivariate analysis, c-MYC amplification was an independent poor-prognostic factor in CRC-S set (P = 0.046). Our study firstly found c-MYC amplification could predict poor prognosis in schistosomiasis-associated colorectal cancer, but not in colorectal cancer without schistosomiasis.


Tyrosinemia type 1: an overview of nursing care.
Tyrosinemia type 1 (TT1) is an inherited metabolic disease that can be fatal when not detected early by newborn screening. In the past, children with TT1 had a poor prognosis due to organ failure and neurologic crisis during infancy. Recent improvements in newborn screening have changed the prognosis of affected children. Measurement of succinylacetone by tandem mass spectrometry provides early identification and the opportunity to manage TT1 as a chronic disease. Treatment includes genetic counseling, dietary management, pharmacotherapy, metabolic crisis prevention, and whole organ transplant. Nursing care is critical to successful management when it is based on a clear understanding of the pathophysiology. This overview of nursing care will provide specific recommendations to reduce complications and enhance the quality of life for children with TT1.


Putative lateral inhibition in sensory processing for directional turns.
Computing targeted responses is a general problem in goal-directed behaviors. We sought the sensory template for directional turning in the predatory sea slug Pleurobranchaea californica, which calculates precise turn angles by averaging multiple stimulus sites on its chemotactile oral veil (Yafremava LS, Anthony CW, Lane L, Campbell JK, Gillette R. J Exp Biol 210: 561-569, 2007). Spiking responses to appetitive chemotactile stimulation were recorded in the two bilateral pairs of oral veil nerves, the large oral veil nerve (LOVN) and the tentacle nerve (TN). The integrative abilities of the peripheral nervous system were significant. Nerve spiking responses to punctate, one-site stimulation of the oral veil followed sigmoid relations as stimuli moved between lateral tentacle and the midline. Receptive fields of LOVN and TN were unilateral, overlapping, and oppositely weighted for responsiveness across the length of oral veil. Simultaneous two-site stimulation caused responses of amplitudes markedly smaller than the sum of corresponding one-site responses. Plots of two-site nerve responses against the summed approximate distances from midline of each site were markedly linear. Thus the sensory paths in the peripheral nervous system show reciprocal occlusion similar to lateral inhibition. This outcome suggests a novel neural function for lateral inhibitory mechanisms, distinct from simple contrast enhancement, in computation of both sensory maps and targeted motor actions.


Diagnosis of asymptomatic primary hyperparathyroidism: proceedings of the Fourth International Workshop.
Asymptomatic primary hyperparathyroidism (PHPT) is a common clinical problem. The purpose of this report is to provide an update on the use of diagnostic tests for this condition in clinical practice. This subgroup was constituted by the Steering Committee to address key questions related to the diagnosis of PHPT. Consensus was established at a closed meeting of the Expert Panel that followed. Each question was addressed by a relevant literature search (on PubMed), and the data were presented for discussion at the group meeting. Consensus was achieved by a group meeting. Statements were prepared by all authors, with comments relating to accuracy from the diagnosis subgroup and by representatives from the participating professional societies. We conclude that: 1) reference ranges should be established for serum PTH in vitamin D-replete healthy individuals; 2) second- and third-generation PTH assays are both helpful in the diagnosis of PHPT; 3) normocalcemic PHPT is a variant of the more common presentation of PHPT with hypercalcemia; 4) serum 25-hydroxyvitamin D concentrations should be measured and, if vitamin D insufficiency is present, it should be treated as part of any management course; 5) genetic testing has the potential to be useful in the differential diagnosis of familial hyperparathyroidism or hypercalcemia.


Nanostructured Biomaterials and Their Applications.
Some of the most important advances in the life sciences have come from transitioning to thinking of materials and their properties on the nanoscale rather than the macro or even microscale. Improvements in imaging technology have allowed us to see nanofeatures that directly impact chemical and mechanical properties of natural and man-made materials. Now that these can be imaged and quantified, substantial advances have been made in the fields of biomimetics, tissue engineering, and drug delivery. For the first time, scientists can determine the importance of nanograins and nanoasperities in nacre, direct the nucleation of apatite and the growth of cells on nanostructured scaffolds, and pass drugs tethered to nanoparticles through the blood-brain barrier. This review examines some of the most interesting materials whose nanostructure and hierarchical organization have been shown to correlate directly with favorable properties and their resulting applications.


Working memory functioning in children with attention-deficit/hyperactivity disorder (ADHD): A comparison between subtypes and normal controls.
Children with attention-deficit/hyperactivity disorder (ADHD) often experience working memory difficulties. However, research findings are inconsistent, making it difficult to compare results across studies. There are several reasons for this inconsistency. Firstly, most studies make no distinction between ADHD subtypes, despite evidence that predominantly inattentive ADHD (ADHD/I) represents a different neurocognitive profile to the hyperactive-impulsive subtype (ADHD/HI). Secondly, documented studies use different tests of working memory which may be measuring different skills. Some assess only the verbal components of working memory and others the visuo-spatial; few of the tests assess both. Further, some tests employ a recognition methodology and others use recall, which require different brain regions and cognitive processes. To clarify these inconsistencies, the verbal and visuo-spatial working memory of children with ADHD/I, ADHD/HI and a control group with no ADHD symptoms were compared. The Automated Working Memory Assessment and Ravens Coloured Progressive Matrices were administered to 72 children (ADHD/I n = 27; ADHD/HI n = 25; control n = 20). The ADHD groups performed similarly, but were significantly poorer than the control group in all aspects of working memory. Storage abilities were stronger than processing abilities, while verbal and visuo-spatial abilities were equally developed for all groups. ADHD-related deficits were apparent across working memory components, even when IQ was controlled for, suggesting a generalised impairment.


Randomized feasibility trial of directly observed versus unobserved hepatitis C treatment with ledipasvir-sofosbuvir among people who inject drugs.
The advent of direct-acting antivirals for hepatitis C virus (HCV) and limited effectiveness of prevention have generated interest in "Treatment as Prevention" (TasP), in which those most likely to transmit HCV (i.e. people who inject drugs [PWID]) are treated to reduced secondary transmission. However, there are scant data regarding the feasibility of treating PWID at high risk for secondary transmission or the optimal approach to treatment delivery. We conducted a 2:1 randomized trial of modified directly-observed (mDOT) versus unobserved HCV treatment with ledipasvir-sofosbuvir daily for 8 weeks among PWID with 36 weeks of follow-up in San Francisco from 2015-2017. We evaluated recruitment-enrollment, treatment completion, end-of-treatment and 12-week response, and reinfection rate. Of 83 individuals eligible for screening, 72 (87.6%) attended the screening visit, 33 were eligible, and 31 enrolled; mean age was 42 years, 81% were male, 74% white. All but one participant (in the mDOT arm) completed treatment and 89.4% of mDOT and 96.6% of unobserved arm visits were attended. HCV was undetectable for 96.8% (30/31) at end of treatment and 89.7% (26/29) 12 weeks later (1 relapse, 1 reinfection), with no differences by arm. Two additional reinfections were subsequently identified, for a reinfection rate of 16.3 (95% CI 5.3-50.5) per 100 person-years of observation. It was feasible to recruit active PWID for HCV treatment and achieve high retention, viral response, and satisfaction with either mDOT or unobserved protocols, supporting treatment of PWID at risk of transmitting HCV to others. The reinfection rate suggests we successfully reached a high-risk population and that successful HCV TasP initiatives may aim to be sufficient in scope to significantly lower prevalence in the community. clinicaltrials.gov NCT02609893.


Integrated approach in the control and management of skin neglected tropical diseases in three health districts of Côte d'Ivoire.
Neglected tropical diseases (NTDs) comprise 20 communicable diseases that are prevalent in rural poor and remote communities with less access to the health system. For effective and efficient control, the WHO recommends that affected countries implement integrated control interventions that take into account the different co-endemic NTDs in the same community. However, implementing these integrated interventions involving several diseases with different etiologies, requiring different control approaches and driven by different vertical programs, remains a challenge. We report here the results and lessons learned from a pilot test of this integrated approach based on integrated screening of skin diseases in three co-endemic health districts of Côte d'Ivoire, a West African country endemic for Buruli ulcer, leprosy and yaw. This cross-sectional study took place from April 2016 to March 2017 in 3 districts of Côte d'Ivoire co-endemic for BU, leprosy and yaws. The study was carried out in 6 stages: identification of potentially co-endemic communities; stakeholder training; social mobilization; mobile medical consultations; case detection and management; and a review meeting. We included in the study all patients with skin signs and symptoms at the screening stage who voluntarily accepted screening. In total, 2310 persons screened had skin lesions at the screening stage. Among them, 07 cases were diagnosed with Buruli ulcer. There were 30 leprosy cases and 15 yaws detected. Other types of ulcerations and skin conditions have been identified and represent the majority of cases detected. We learned from this pilot experience that integration can be successfully implemented in co-endemic communities in Côte d'Ivoire. Health workers are motivated and available to implement integrated interventions instead of interventions focused on a single disease. However, it is essential to provide capacity building, a minimum of drugs and consumables for the care of the patients identified, as well as follow-up of identified patients, including those with other skin conditions. The results of this study show that the integration of activities can be successfully implemented in co-endemic communities under the condition of staff capacity building and minimal care of identified patients.


Ultracytochemical study of ouabain-sensitive, potassium-dependent p-nitrophenylphosphatase activity in the inner ear of the squirrel monkey.
The localization of ouabain-sensitive, K+-dependent p-nitrophenylphosphatase (K+-NPPase) activity of the Na+, K+-ATPase complex was studied ultracytochemically in the squirrel monkey inner ear. In the stria vascularis the reaction products showing K+-NPPase activity were limited to the cytoplasmic side of the plasmalemmal infoldings of the marginal cells. In the spiral prominence, a weak reaction was also found on the cytoplasmic process of the stromal cell, while no or little reaction was detected on the spiral prominence epithelium. In the dark cells of vestibular labyrinth the reaction products were observed on the basolateral interdigitation of the plasmalemma. In contrast, no reaction was observed on the apical cell surface. K+-NPPase activity was most intense in the strial marginal cell, followed by the dark cell of the ampulla and the utricle. The present results revealed that the dark cells in the vestibular labyrinth are involved in endolymph homeostasis.


Treatment Outcome of the Drug-resistant Zoonotic Cutaneous Leishmaniasis by Glucantime.
Resistance of Leishmania species to antimonial drugs has increased. Hence, in the present study Leishmania major isolates were collected from patients with resistance phenotype and the presence/absence of resistance to Glucantime was investigated. Samples were taken from 10 cutaneous leishmaniasis (CL) patients who had not responded to chemotherapy with Glucantime. Nested polymerase chain reaction (PCR) was performed to identify the isolated species. Stationary phase promastigotes were added to the grown, adhesive J774 macrophages. Values obtained from standard strain were compared with the test cultures after exposure to the medicine. In vivo, the effects of Glucantime were assessed by comparing the sizes and the parasite burden of the lesions on mouse model. The results of amplified band on agarose gel demonstrated all samples were L. major. After exposure to medicine, a reduction of intracellular amastigotes to half was detected. In vivo, the parasite was eliminated in 90% of mice with lesions caused by both isolates of patients and standard L. major, and their lesions became smaller significantly. Pentavalent antimonial (SbV) salts are the main component of chemotherapy against leishmaniasis. However, the medicine has been found ineffective. In the present study, isolates from patients with no response to treatment had no significant difference from the standard L. major strain (as the sensitive strain). Therefore, in patients with resistance phenotype to Glucantime, the parasites did not actually have intrinsic resistance, i.e., environmental and host factors prevented the successful treatment of the disease.


Isolated atherosclerotic aneurysms of the internal iliac arteries: report of two cases and review of literature.
Isolated arteriosclerotic aneurysms of the internal iliac artery are uncommon, with an incidence approaching 0.4%. Although the symptoms invariably involve the genitourinary or gastrointestinal systems, the aneurysms may be found during abdominal, rectal, or vaginal examinations. The 75% mortality associated with the rupture of an internal iliac aneurysm mandates for early surgery for these aneurysms. Operation may consist of ligation of the internal iliac artery, excision of the aneurysm, or an obliterative endoaneurysmorrhaphy. While the operative mortality has average 33.3%, there were no operative deaths in this series.


The science of neonatal high-frequency ventilation.
Despite improvements in respiratory care, ventilator-induced lung injury remains an important cause of morbidity and mortality in neonates who require assisted ventilation. Animal data clearly demonstrate that high-frequency ventilation can be used successfully to reduce lung injury in experimental models of acute lung injury. These models and human research show that the efficacy of high-frequency ventilation is dependent on optimizing functional residual capacity and avoiding lung overinflation. When used with a strategy that promotes lung recruitment, high-frequency ventilation effectively reduces the occurrence of chronic lung disease and is not associated with significant brain injury. When used with a strategy that allows the lung to collapse or is associated with hyperventilation, however, high-frequency ventilation does not reduce lung injury and is associated with significant brain injury. Like every tool we use to support critically ill neonates, high-frequency ventilation needs a careful carpenter. As therapies and health care strategies evolve, there remains nothing more important than the health care team at the bedside. Critical evaluation of the patient and his or her response to the therapy being offered is essential to promotion of the patient health outcome.


Carbonyl Olefin Metathesis and Dehydrogenative Cyclization of Aromatic Ketones and gem-Difluoroalkenes.
The beauty of one-pot cascade reaction lies in the efficient disconnection and construction of several bonds in a single reaction flask, without the isolation of any intermediates. Herein, we report the first photo-induced thermal-promoted cascade reactions of readily available aromatic ketones and aromatic gem-difluoroalkenes for the synthesis of phenanthrenes which possess potential utility in drug design and material science. The reaction combines carbonyl-olefin metathesis (cascade photo-induced [2+2] cyclization and thermal-controlled retro [2+2] cyclization) and dehydrogenative cyclization (cascade photo-induced conrotatory 6π electrocyclization and collidine-promoted dehydrogenative aromatization) together in one pot. The oxidant-free, acid-free and metal-free reaction shows broad substrate scope and wide functional group tolerance.


Analysis of DNA restriction fragments and polymerase chain reaction products towards detection of the AIDS (HIV-1) virus in blood.
A high-performance capillary electrophoresis system with a polysiloxane-coated capillary and polymeric buffer additives was investigated for the analysis of DNA restriction fragments and polymerase chain reaction (PCR) products. Mobility data and Ferguson plots of the DNA fragments at different polymer (hydroxypropylmethylcellulose) concentrations indicated that effective molecular sieving was obtained consistent with existing data of conventional gel electrophoresis and with recent HPCE data. The precision and peak efficiency were excellent and the system was applied to the analysis of specific co-amplified DNA sequences (HIV-1 and HLA-DQ-alpha). After PCR, ultrafiltration was used in the sample preparation step to desalt the sample and to remove superfluous PCR reaction products. Electrokinetic injection was used for sample introduction into the capillary. The addition of ethidium bromide to the buffer resulted in longer migration times of DNA fragments and better peak resolution. During HPCE, an artifact associated with dilute DNA solutions leading to the appearance of extra peaks in the electropherogram was found.


Effect of dietary salt intake on epithelial Na+ channels (ENaC) in vasopressin magnocellular neurosecretory neurons in the rat supraoptic nucleus.
A growing body of evidence suggests that epithelial Na+ channels (ENaCs) in the brain play a significant role in the regulation of blood pressure; however, the brain structures that mediate the effect are not well understood. Because vasopressin (VP) neurons play a pivotal role in coordinating neuroendocrine and autonomic responses to maintain cardiovascular homeostasis, a basic understanding of the regulation and activity of ENaC in VP neurons is of great interest. We show that high dietary salt intake caused an increase in the expression and activity of ENaC which resulted in the steady state depolarization of VP neurons. The results help us understand one of the mechanisms underlying how dietary salt intake affects the activity of VP neurons via ENaC activity. All three epithelial Na+ channel (ENaC) subunits (α, β and γ) are located in vasopressin (VP) magnocellular neurons in the hypothalamic supraoptic (SON) and paraventricular nuclei. Our previous study demonstrated that ENaC mediates a Na+ leak current that affects the steady state membrane potential in VP neurons. In the present study, we evaluated the effect of dietary salt intake on ENaC regulation and activity in VP neurons. High dietary salt intake for 7 days caused an increase in expression of β- and γENaC subunits in the SON and the translocation of αENaC immunoreactivity towards the plasma membrane. Patch clamp experiments on hypothalamic slices showed that the mean amplitude of the putative ENaC currents was significantly greater in VP neurons from animals that were fed a high salt diet compared with controls. The enhanced ENaC current contributed to the more depolarized basal membrane potential observed in VP neurons in the high salt diet group. These findings indicate that high dietary NaCl intake enhances the expression and activity of ENaCs, which augments synaptic drive by depolarizing the basal membrane potential close to the action potential threshold during hormonal demand. However, ENaCs appear to have only a minor role in the regulation of the firing activity of VP neurons in the absence of synaptic inputs as neither the mean intraburst frequency, burst duration, nor interspike interval variability of phasic bursting activity was affected. Moreover, ENaC activity did not affect the initiation, sustention, or termination of the phasic bursting generated in an intrinsic manner without synaptic inputs.


Several thaumatin-like proteins bind to beta-1,3-glucans.
Pathogenesis-related proteins from intercellular fluid washings of stressed barley (Hordeum vulgare L.) leaves were analyzed to determine their binding to various water-insoluble polysaccharides. Three proteins (19, 16, and 15 kD) bound specifically to several water-insoluble beta-1,3-glucans. Binding of the barley proteins to pachyman occurred quickly at 22 degreesC at pH 5.0, even in the presence of 0.5 M NaCl, 0.2 M urea, and 1% (v/v) Triton X-100. Bound barley proteins were released by acidic treatments or by boiling in sodium dodecyl sulfate. Acid-released barley proteins could bind again specifically and singly to pachyman. Water-soluble laminarin and carboxymethyl-pachyman competed for the binding of the barley proteins to pachyman. The N-terminal sequence of the 19-kD barley beta-1,3-glucan-binding protein showed near identity to the barley seed protein BP-R and high homology to other thaumatin-like (TL) permatins. The 16-kD barley protein was also homologous to TL proteins, whereas the 15-kD barley protein N-terminal sequence was identical to the pathogenesis-related Hv-1 TL protein. Antifungal barley protein BP-R and corn (Zea mays) zeamatin were isolated by binding to pachyman. Two extracellular proteins from stressed pea (Pisum sativum L.) also bound to pachyman and were homologous to TL proteins.


Orientation anisotropy: incidence and magnitude in Caucasian and Chinese subjects.
Cutoff spatial frequencies for sine-wave gratings were measured at four orientations for 100 Caucasian and 24 Chinese subjects, all of whom were raised in carpentered enviornments. For the Caucasian subjects, average acuity for horizontal and vertical gratings was superior to that for obliques by about one-quarter of an octive. However, about 10 percent of subjects showed an anisotropy of about one-eight of an ovtave. The carpentered enviornment explanation of orientation anisotropy cannot, in its present form, account for the wide variety of response patterns obtained, nor the differences between Chinese and Caucasian subjects.


Extraction and analysis of carotenoids from the New Zealand sea urchin Evechinus chloroticus gonads.
Sea urchin gonad (roe) is a highly valued food in Japan and North America. Gonad price is strongly influenced by quality, with appearance, especially colour being a major determinant. Previous attempts to extract a carotenoid profile from the New Zealand sea urchin species Evechinus chloroticus have been challenging due to the large amount of lipid present in the gonad. A carotenoid extraction and high performance liquid chromatography (HPLC) analysis method was developed to reduce lipid contamination by incorporating a saponification and lipid cold precipitation in the extraction procedure. This method enabled greater carotenoid purity and enhanced analysis by HPLC. Echinenone was found to be the main carotenoid present in all E. chloroticus gonads. Dark coloured gonads contained higher levels of fucoxanthin/fucoxanthinol, β-carotene and xanthophylls such as astaxanthin and canthaxanthin. This information on the modification and deposition of carotenoids will help in the development of diets to enhance gonad colour.


Analysis of natural allelic variation at flowering time loci in the Landsberg erecta and Cape Verde Islands ecotypes of Arabidopsis thaliana.
We have analyzed the flowering behavior of two Arabidopsis ecotypes: the laboratory strain Landsberg erecta (Ler) and an ecotype from the tropical Cape Verde Islands (Cvi). They differ little in their flowering phenotypes and in their responses to photoperiod length changes and to vernalization treatment. However, segregating populations derived from crosses between them showed a much larger variation. An approach of quantitative trait locus (QTL) mapping in recombinant inbred lines (RILs) grown under three environments differing in day-length and/or vernalization treatment has been used to detect and locate flowering loci. Four main QTLs were identified, designated early day-length insensitive (EDI), flowering F, G, and H (FLF, FLG, and FLH, respectively), to which most of the flowering behavior differences could be attributed. To further characterize the individual loci, near isogenic lines were constructed by introgressing Cvi early alleles of EDI and FLH into the Ler genetic background. EDI-Cvi alleles produce earliness under both long- and short-day photoperiods, rendering Ler plants almost day-length neutral. In addition, RILs were selected to analyze FLF and FLG. These loci interact epistatically and RILs carrying late alleles at FLF and FLG were very responsive to vernalization and showed an increased response to photoperiod length changes. The possible role of these loci for the control of flowering is discussed in the context of the current Arabidopsis model.