Breast cancer in women: a descriptive analysis of the national cancer database.
Breast cancer is the most common cancer in women in the United States. National Cancer Database (NCDB) is one of the largest tumor databases of the United States. This study aimed to evaluate the features of breast cancer in women from a large updated database. We describe and analyze the frequencies and percentages of the clinical and pathological features of women diagnosed with breast cancer registered in NCDB, in a period from 2004 to 2015. A total of 2,423,875 women were diagnosed with breast cancer between 2004 and 2015. The nationally representative analysis demonstrated that the incidence of breast cancer among women increased over the years. Upper-outer quadrant was the most frequent primary tumor site, and the intraductal carcinoma was the most frequent histology. The prevalence of breast cancer increased with age. The most frequent grade at diagnosis was grade II. Broadly, invasive characteristics were noted more frequently in younger patients. Left and right breast were affected with almost the same frequency, with a slight predominance of the left breast. The most frequent surgical treatment was a partial mastectomy. Reconstruction with implant was the most frequent choice. Post-mastectomy radiation therapy was administered in the majority of patients. To the authors' knowledge, the current study is the largest descriptive analysis to date on the clinical and pathological features of breast cancer in a population-based database. The increase in incidence over the years indicates an important need for etiologic research and innovative approaches to improve breast cancer prevention.


[Nurses' perception about diseases that are treated by acupuncture].
The objective of this study was to identify and analyze nurses' perceptions about diseases that are treated by acupuncture, taking into consideration: (1) the diseases of nurses who had received acupuncture treatment; (2) nurses' diseases which might be treated by acupuncture; (3) suggestions of diseases treatable by acupuncture. This exploratory study was performed with 33 nurses working at 11 health units in southeast São Paulo; places in which acupuncture treatment was available. Results showed that acupuncture was mostly used for stress (10.7%), anxiety, migraines, lumbar pain, myoma and obesity (7.1% each). Acupuncture was suggested as treatments mostly for musculoskeletal diseases (42%), and chronic-degenerative diseases (10%).


Rifampicin-resistant and rifabutin-susceptible Mycobacterium tuberculosis strains: a breakpoint artefact?
It has long been assumed that some rifampicin-resistant Mycobacterium tuberculosis strains are susceptible to, and thus treatable with, rifabutin. However, clinical breakpoints for susceptibility testing of rifabutin as well as the evidence for a clinical effect of rifabutin in rifampicin-resistant strains remains poorly defined. The objective of this study was to re-evaluate the breakpoint for rifabutin in relation to its MIC wild-type distribution and the presence of mutations in rpoB. The MIC in 7H10 Middlebrook medium was determined for clinical isolates of M. tuberculosis (n = 95), where a majority were multidrug resistant. Additionally, all strains were screened for rpoB mutations by sequencing and the GenoType MTBDRplus assay. Rifampicin resistance was confirmed by genotypical and/or phenotypical tests in 73 isolates (76.8%). Nineteen isolates, defined as rifampicin resistant and rifabutin susceptible according to the present breakpoint, exhibited significantly higher MICs of rifabutin (0.064-0.5 mg/L) than rifabutin-susceptible isolates without any detectable mutations in rpoB (P < 0.001). These 19 isolates were clearly resistant to rifampicin (MIC 2-256 mg/L) and all but one had mutations in rpoB, with 9 (47.4%) specifically in Asp516Val. Our results indicate that rifampicin-resistant but rifabutin-susceptible isolates according to the present breakpoints harbour rpoB mutations and have a rifabutin MIC significantly higher than strains without any detectable mutations in rpoB. So far there are no clinical, pharmacological or microbiological data to confirm that such isolates can be treated with rifabutin and we suggest a revision of the current breakpoints.


Stomach anatomy and use in defining systemic relationships of the Cetacean family Ziphiidae (beaked whales).
Members of the Cetacean family Ziphiidae (Beaked whales) exhibit surprising diversity in stomach anatomy, particularly in the number of connecting chambers between the main and pyloric stomachs. Sixty-one stomachs of five beaked whale species were examined, including Berardius, Hyperoodon, Mesoplodon, Tasmacetus, and Ziphius. Specimens were obtained post mortem from beach-stranded individuals, with the exception of 30 Berardius specimens that were taken at a whaling station in Japan. Most specimens were collected by the Marine Mammal Program of the National Museum of Natural History, with the exception of one specimen of Mesoplodon steinegeri, which from the Los Angeles County Museum and one specimen of Tasmacetus from the Museum of New Zealand. Additional data are included from published accounts. A single species was examined for all of the genera but Mesoplodon, where there was material for 7 of 13 known species. The sample of Berardius bairdii was sufficient (n = 30) to determine species-specific variation in compartment counts. Chamber anatomy was explored by means of manual palpation. Results indicate that ziphiid stomachs can be separated into at least three principal groups: generalized ziphiid stomach (one main stomach, one pyloric stomach), derived stomach type I (two main stomachs, one pyloric stomach), and derived stomach type II (two main stomachs, two pyloric stomachs). Generalized stomachs are found in Hyperoodon, Tasmacetus, Ziphius, Mesoplodon densirostris, M. perrini, and M. stejnegeri. Derived stomachs of type I are found in Berardius, and of type II are found in Mesoplodon bidens, M. europaeus, and M. mirus. The ziphiids clearly form a distinct group of cetaceans in their utilization of differences in stomach morphology. These anatomical differences may serve to elucidate systematic relationships among the ziphiids. Further study is necessary to establish whether these differences correlate with specialized adaptations related to an aquatic environment.


Proton-detected solid-state NMR spectroscopy at aliphatic sites: application to crystalline systems.
When applied to biomolecules, solid-state NMR suffers from low sensitivity and resolution. The major obstacle to applying proton detection in the solid state is the proton dipolar network, and deuteration can help avoid this problem. In the past, researchers had primarily focused on the investigation of exchangeable protons in these systems. In this Account, we review NMR spectroscopic strategies that allow researchers to observe aliphatic non-exchangeable proton resonances in proteins with high sensitivity and resolution. Our labeling scheme is based on u-[(2)H,(13)C]-glucose and 5-25% H2O (95-75% D2O) in the M9 bacterial growth medium, known as RAP (reduced adjoining protonation). We highlight spectroscopic approaches for obtaining resonance assignments, a prerequisite for any study of structure and dynamics of a protein by NMR spectroscopy. Because of the dilution of the proton spin system in the solid state, solution-state NMR (1)HCC(1)H type strategies cannot easily be transferred to these experiments. Instead, we needed to pursue ((1)H)CC(1)H, CC(1)H, (1)HCC or ((2)H)CC(1)H type experiments. In protonated samples, we obtained distance restraints for structure calculations from samples grown in bacteria in media containing [1,3]-(13)C-glycerol, [2]-(13)C-glycerol, or selectively enriched glucose to dilute the (13)C spin system. In RAP-labeled samples, we obtained a similar dilution effect by randomly introducing protons into an otherwise deuterated matrix. This isotopic labeling scheme allows us to measure the long-range contacts among aliphatic protons, which can then serve as restraints for the three-dimensional structure calculation of a protein. Due to the high gyromagnetic ratio of protons, longer range contacts are more easily accessible for these nuclei than for carbon nuclei in homologous experiments. Finally, the RAP labeling scheme allows access to dynamic parameters, such as longitudinal relaxation times T1, and order parameters S(2) for backbone and side chain carbon resonances. We expect that these measurements will open up new opportunities to obtain a more detailed description of protein backbone and side chain dynamics.


Epidermal growth factor receptor expression in breast cancer association with biologic phenotype and clinical outcomes.
Epidermal growth factor receptor (EGFR) expression is associated with aggressive phenotypes in preclinical breast cancer models, but in clinical studies, EGFR has been inconsistently linked to poor outcome. We hypothesized that EGFR expression in human breast tumors, when centrally and uniformly assessed, is associated with an aggressive phenotype and resistance to systemic therapy. In a database of 47,286 patients with breast cancer, EGFR status was known on 2567 tumors. EGFR levels were measured centrally by ligand binding assay, and tumors with > or =10 fmol/mg were prospectively deemed positive. Clinical and biological features of EGFR-positive and EGFR-negative tumors were compared. Clinical outcomes were assessed by systemic therapy status. Of 2567 tumors, 475 (18%) were EGFR positive. EGFR-positive tumors were more common in younger and in black women, were larger, had a higher S-phase fraction, and were more likely to be aneuploid. EGFR-positive tumors were more likely to be HER2-positive (26% vs 16%, P < .0001), but less likely to be estrogen receptor-positive (60% vs 88%, P < .0001) or progesterone receptor-positive (26% vs 65%, P < .0001). In multivariate analyses, EGFR expression independently correlated with worse disease-free survival (hazard ratio [HR] = 1.66; 95% confidence interval [CI], 1.4-2.41, P = .007) and overall survival (HR = 1.98, 95% CI, 1.36-2.88, P = .0004) in treated patients, but not in untreated patients. EGFR expression is more common in breast tumors in younger and black women. It is associated with lower hormone receptor levels, higher proliferation, genomic instability, and HER2 overexpression. It is correlated with higher risk of relapse in patients receiving adjuvant treatment. Blocking EGFR may improve outcome in selected patients.


Combined PET and microdialysis for in vivo estimation of drug blood-brain barrier transport and brain unbound concentrations.
Methods to investigate blood-brain barrier transport and pharmacologically active drug concentrations in the human brain are limited and data translation between species is challenging. Hence, there is a need to further develop the read-out of techniques like positron emission tomography (PET) for studying neuropharmacokinetics. PET has a high translational applicability from rodents to man and measures total drug concentrations in vivo. The aim of the present study was to investigate the possibility of translating total drug concentrations, acquired through PET, to unbound concentrations, resembling those measured in the interstitial fluid by microdialysis sampling. Simultaneous PET scanning and brain microdialysis sampling were performed in rats throughout a 60min infusion of [N-methyl-11C]oxycodone in combination with a therapeutic dose of oxycodone and during a 60min follow up period after the end of infusion. The oxycodone concentrations acquired with PET were converted into unbound concentrations by compensating for brain tissue binding and brain intracellular distribution, using the unbound volume of distribution in brain (Vu,brain), and were compared to microdialysis measurements of unbound concentrations. A good congruence between the methods was observed throughout the infusion. However, an accumulating divergence in the acquired PET and microdialysis data was apparent and became more pronounced during the elimination phase, most likely due to the passage of radioactive metabolites into the brain. In conclusion, the study showed that PET can be used to translate non-invasively measured total drug concentrations into unbound concentrations as long as the contribution of radiolabelled metabolites is minor or can be compensated for.


Automatic characterization of soft tissues material properties during mechanical tests.
The estimation of the non-linear viscoelastic characteristics of human soft tissues, such as ligaments and tendon, is often affected by the implemented procedure. This study aims at developing and validating a protocol, associated with a contactless and automatic procedure, enabling the determination of the material behavior and properties of any soft tissues. Several markers were drawn onto the soft tissue specimen analyzed under uniaxial tensile test. An automatic contactless procedure, that uses a camera for recording the position of the markers during the test, was developed to compute the displacement, and the force applied, enabling the calculation of the true-stress/strain curve of the material. Young's modulus and Poisson's ratio can be calculated, on demand, for selected regions of interest of the soft tissues. The repeatability and reproducibility of the procedure were analyzed. The procedure was initially tested and verified on an artificial silicone material and later applied for investigating the mechanical behavior of a pig Achilles tendon and of a human patellar tendon. The procedure show a high repeatability, independent by the operator, reliability and accuracy for the tested synthetic material (with a maximum error of 3.7% for Young's modulus). Additionally, the developed protocol was also suitable for the analysis of animal and human soft tissues. A protocol to automatically and accurately determine material properties in soft tissues was developed, tested and validated. Such approach could successfully be implemented for the mechanical characterization of any biological soft-tissue. V.


Development of nursing documentation for use in the outpatient oncology setting.
In response to the particular demands upon patients and nursing staff in the outpatient setting, and the identification of a need for nursing orientated documentation, a new nursing documentation system has been developed and implemented within an oncology day unit. The documentation is used by the nursing team engaged in the care of patients receiving outpatient chemotherapy treatment. The system consists of documents for the initial and ongoing assessment, planning, implementation and evaluation of nursing care. These include pretreatment patient self-assessment forms, flowsheets for the documentation of chemotherapy treatment and administration and a record for information and teaching given to patients and significant others. Referrals to other services and health care professionals are also documented. This tool is designed to ensure a high standard of nursing intervention and maximize quality of life in this group of patients. An audit is planned to evaluate the effectiveness of this tool and highlight any areas requiring improvement or modification. There are also plans to further develop this system for use in the inpatient and community setting.


Glycosylation-dependent activation of epithelial sodium channel by solnatide.
Dysfunction of the epithelial sodium channel (ENaC), which regulates salt and water homeostasis in epithelia, causes several human pathological conditions, including pulmonary oedema. This is a potentially lethal complication of acute lung injury at least partially caused by dysfunctional alveolar liquid clearance, which in turn impairs alveolar gas exchange. Solnatide (named TIP-peptide, AP301), a 17 residue peptide mimicking the lectin-like domain of TNF has been shown to activate ENaC in several experimental animal models of acute lung injury and is being evaluated as a potential therapy for pulmonary oedema. The peptide has recently completed phase 1 and 2a clinical trials. In this study, we identify a glycosylation-dependent mechanism that preserves ENaC function and expression. Since our previous data suggested that the pore-forming subunits of ENaC are essential for maximal current activation by solnatide, we performed single- and multi-N-glycosylation site mutations in αN232,293,312,397,511Q- and δN166,211,384Q-subunits, in order to identify crucial residues for interaction with solnatide within the extracellular loop of the channel. Additionally, we generated αL576X and αN232,293,312,397,511Q,L576X deletion mutants of ENaC-α, since we have previously demonstrated that the carboxy terminal domain of this subunit is also involved in its interaction with solnatide. In cells expressing αN232,293,312,397,511Q,L576Xβγ-hENaC or δN166,311,384Q,D552Xβγ-hENaC activation by solnatide, as measured in whole cell patch clamp mode, was completely abolished, whereas it was attenuated in αL576Xβγ-hENaC- and δD552Xβγ-hENaC-expressing cells. Taken together, our findings delineate an N-glycan dependent interaction between the TIP-peptide and ENaC leading to normalization of both sodium and fluid absorption in oedematous alveoli to non-oedematous levels.


Decreased infectivity following BNT162b2 vaccination: A prospective cohort study in Israel.
BNT162b2 was shown to be 92% effective in preventing COVID-19. Prioritizing vaccine rollout, and achievement of herd immunity depend on SARS-CoV-2 transmission reduction. The vaccine's effect on infectivity is thus a critical priority. Among all 9650 HCW of a large tertiary medical center in Israel, we calculated the prevalence of positive SARS-CoV-2 qRT-PCR cases with asymptomatic presentation, tested following known or presumed exposure and the infectious subset (N-gene-Ct-value<30) of these. Additionally, infection incidence rates were calculated for symptomatic cases and infectious (Ct<30) cases. Vaccine effectiveness within three months of vaccine rollout was measured as one minus the relative risk or rate ratio, respectively. To further assess infectiousness, we compared the mean Ct-value and the proportion of infections with a positive SARS-CoV-2 antigen test of vaccinated vs. unvaccinated. The correlation between IgG levels within the week before detection and Ct level was assessed. Reduced prevalence among fully vaccinated HCW was observed for (i) infections detected due to exposure, with asymptomatic presentation (VE(i)=65.1%, 95%CI 45-79%), (ii) the presumed infectious (Ct<30) subset of these (VE(ii)=69.6%, 95%CI 43-84%) (iii) never-symptomatic infections (VE(iii)=72.3%, 95%CI 48-86%), and (iv) the presumed infectious (Ct<30) subset (VE(iv)=83.0%, 95%CI 51-94%).Incidence of (v) symptomatic and (vi) symptomatic-infectious cases was significantly lower among fully vaccinated vs. unvaccinated individuals (VE(v)= 89.7%, 95%CI 84-94%, VE(vi)=88.1%, 95%CI 80-95%).The mean Ct-value was significantly higher in vaccinated vs. unvaccinated (27.3±1.2 vs. 22.2±1.0, p<0.001) and the proportion of positive SARS-CoV-2 antigen tests was also significantly lower among vaccinated vs. unvaccinated PCR-positive HCW (80% vs. 31%, p<0.001). Lower infectivity was correlated with higher IgG concentrations (R=0.36, p=0.01). These results suggest that BNT162b2 is moderately to highly effective in reducing infectivity, via preventing infection and through reducing viral shedding. Sheba Medical Center, Israel.


Understanding Variation in Rotavirus Vaccine Effectiveness Estimates in the United States: The Role of Rotavirus Activity and Diagnostic Misclassification.
Estimates of rotavirus vaccine effectiveness (VE) in the United States appear higher in years with more rotavirus activity. We hypothesized rotavirus VE is constant over time but appears to vary as a function of temporal variation in local rotavirus cases and/or misclassified diagnoses. We analyzed 6 years of data from eight US surveillance sites on 8- to 59-month olds with acute gastroenteritis symptoms. Children's stool samples were tested via enzyme immunoassay (EIA); rotavirus-positive results were confirmed with molecular testing at the US Centers for Disease Control and Prevention. We defined rotavirus gastroenteritis cases by either positive on-site EIA results alone or positive EIA with Centers for Disease Control and Prevention confirmation. For each case definition, we estimated VE against any rotavirus gastroenteritis, moderate-to-severe disease, and hospitalization using two mixed-effect regression models: the first including year plus a year-vaccination interaction, and the second including the annual percent of rotavirus-positive tests plus a percent positive-vaccination interaction. We used multiple overimputation to bias-adjust for misclassification of cases defined by positive EIA alone. Estimates of annual rotavirus VE against all outcomes fluctuated temporally, particularly when we defined cases by on-site EIA alone and used a year-vaccination interaction. Use of confirmatory testing to define cases reduced, but did not eliminate, fluctuations. Temporal fluctuations in VE estimates further attenuated when we used a percent positive-vaccination interaction. Fluctuations persisted until bias-adjustment for diagnostic misclassification. Both controlling for time-varying rotavirus activity and bias-adjusting for diagnostic misclassification are critical for estimating the most valid annual rotavirus VE.


Nanoscale neuro-integrative coatings for neural implants.
Silicon microelectrode arrays (Si MEAs) have great potential in enabling chronic in vivo recording of neural activity, but this potential has been hampered by scar tissue formation at the site of implantation. In this study, we report the fabrication and characterization of nanoscale coatings that have the potential of enhancing the biocompatibility of Si electrodes. We use electrostatic layer-by-layer (LbL) assembly to prepare nanoscale bioactive coatings on silicon substrates. We use the response of chick cortical neurons to these coatings to assess potential improvement in biocompatibility in vitro. The coatings are built on oxide covered silicon wafers by alternating polycations, polyethyleneimine (PEI) or chitosan (CH), with polyanions, either gelatin or laminin (LN). We use quartz crystal microbalance (QCM) to characterize the coatings. Our analysis confirms that we achieved approximately 30-110 angstroms scale coatings via LbL assembly. In contrast to bare oxide covered silicon, coated substrates had significantly enhanced chick cortical neuron adhesion and differentiation, with multilayers of PEI-LN showing the greatest improvement. The multilayers of PEI-LN were stable for at least 7 days in physiological conditions, as determined by an enzyme-linked immunosorbent assay (ELISA). In addition, impedance spectroscopy confirmed that multilayers of PEI and LN did not increase the magnitude of impedance of Si MEAs at the biologically relevant frequency of 1 kHz. Our study demonstrates that electrostatic LbL assembly enables nanoscale bioactive coatings, and that PEI-LN multilayers significantly enhance cortical neuronal attachment and differentiation in vitro with no deleterious effects on impedance of the electrodes. Such well-controlled nanoscale coatings have the potential to significantly impact the compatibility and performance of Si MEAs in vivo.


Ontogeny of ependymoglial cells lining the third ventricle in mice.
During hypothalamic development, the germinative neuroepithelium gives birth to diverse neural cells that regulate numerous physiological functions in adulthood. Here, we studied the ontogeny of ependymal cells in the mouse mediobasal hypothalamus using the BrdU approach and publicly available single-cell RNAseq datasets. We observed that while typical ependymal cells are mainly produced at E13, tanycyte birth depends on time and subtypes and lasts up to P8. Typical ependymocytes and β tanycytes are the first to arise at the top and bottom of the dorsoventral axis around E13, whereas α tanycytes emerge later in development, generating an outside-in dorsoventral gradient along the third ventricle. Additionally, α tanycyte generation displayed a rostral-to-caudal pattern. Finally, tanycytes mature progressively until they reach transcriptional maturity between P4 and P14. Altogether, this data shows that ependyma generation differs in time and distribution, highlighting the heterogeneity of the third ventricle.


Perfect adaptation achieved by transport limitations governs the inorganic phosphate response in S. cerevisiae.
Cells cope with and adapt to ever-changing environmental conditions. Sophisticated regulatory networks allow cells to adjust to these fluctuating environments. One such archetypal system is the Saccharomyces cerevisiae Pho regulon. When external inorganic phosphate (Pi) concentration is low, the Pho regulon activates, expressing genes that scavenge external and internal Pi. However, the precise mechanism controlling this regulon remains elusive. We conducted a systems analysis of the Pho regulon on the single-cell level under well-controlled environmental conditions. This analysis identified a robust, perfectly adapted Pho regulon state in intermediate Pi conditions, and we identified an intermediate nuclear localization state of the transcriptional master regulator Pho4p. The existence of an intermediate nuclear Pho4p state unifies and resolves outstanding incongruities associated with the Pho regulon, explains the observed programmatic states of the Pho regulon, and improves our general understanding of how nature evolves and controls sophisticated gene regulatory networks. We further propose that robustness and perfect adaptation are not achieved through complex network-centric control but by simple transport biophysics. The ubiquity of multitransporter systems suggests that similar mechanisms could govern the function of other regulatory networks as well.


Slotted photonic crystal sensors.
Optical biosensors are increasingly being considered for lab-on-a-chip applications due to their benefits such as small size, biocompatibility, passive behaviour and lack of the need for fluorescent labels. The light guiding mechanisms used by many of them results in poor overlap of the optical field with the target molecules, reducing the maximum sensitivity achievable. This review article presents a new platform for optical biosensors, namely slotted photonic crystals, which provide higher sensitivities due to their ability to confine, spatially and temporally, the optical mode peak within the analyte itself. Loss measurements showed values comparable to standard photonic crystals, confirming their ability to be used in real devices. A novel resonant coupler was designed, simulated, and experimentally tested, and was found to perform better than other solutions within the literature. Combining with cavities, microfluidics and biological functionalization allowed proof-of-principle demonstrations of protein binding to be carried out. Higher sensitivities were observed in smaller structures than possible with most competing devices reported in the literature. This body of work presents slotted photonic crystals as a realistic platform for complete on-chip biosensing; addressing key design, performance and application issues, whilst also opening up exciting new ideas for future study.


The anti-obesity effect of starch in a whole grain-like structural form.
Obesity is a risk factor for many chronic diseases, and the anti-obesity effect of starch in a whole grain-like structural form (WGLSF) prepared through co-gelation with oat β-glucan and alginate was studied using high-fat (HF) induced obese male C57BL/6J mice. In vitro human fecal fermentation of WGLSF-starch showed a slower rate of fermentation and a higher production of butyric acid (132.0 μmol per 50 mg sample) when compared to the physical mixture counterpart of starch, β-glucan, and alginate (PM) (110.5 μmol per 50 mg) or β-glucan itself (96.2 μmol per 50 mg). The body weight gain of obese mice fed with a HF-WGLSF diet was significantly reduced (42.0% lower than the HF group, 30.2% lower than the physical mixture) with decreased cell size in white adipose tissue and similar levels of serum lipid profiles to the control of the low-fat (LF) group. Western blotting experiments showed the down-regulated lipogenic transcription factor of SREBP-1c and fatty acid synthase (FAS), but the lipid-oxidation related transcription factors of peroxisome proliferator-activated receptor-α (PPAR-α) and phosphorylated AMP-activated protein kinase (p-AMPK) were up-regulated. Energy metabolism analysis revealed increased lipid-sourced energy expenditure with higher heat production and respiratory exchange ratios. Consistently, the expression of hypothalamic pro-opiomelanocortin (POMC), favoring energy expenditure, was increased significantly while the neuropeptide Y (NPY) was reduced. Thus, the increased energy expenditure stimulated by starch in a whole-grain-like structural form is responsible for the reduced body weight gain of obese mice fed with a high fat-based diet.


[Study of leprosy at the "Emilio d'Audinot" polyclinic].
A study on the prevalence of leprosy is performed in "Emilio Daudinot" polyclinics, Guantanamo. The behaviour of leprosy according to clinical manifestations and results showing that lepromatous leprosy is the most frequent form, as well as the number of positive patients and the ones who present reactional status, are reported. The clinical course according to skin and general manifestations is analyzed, and it was determined that the greatest number of cases improved with treatment. Likewise, it was found that the greatest number of positive patients require a 1-4 year period for becoming negative. The immunologic status of cases studied according to the different clinical forms is reported and it was observed that the greatest number of cases are immunologically depressed which agrees with the fact that most of patients have the lepromatous form. Nurses play a very important role for the control and treatment of these patients.


Artificial intelligence in molecular imaging.
AI has, to varying degrees, affected all aspects of molecular imaging, from image acquisition to diagnosis. During the last decade, the advent of deep learning in particular has transformed medical image analysis. Although the majority of recent advances have resulted from neural-network models applied to image segmentation, a broad range of techniques has shown promise for image reconstruction, image synthesis, differential-diagnosis generation, and treatment guidance. Applications of AI for drug design indicate the way forward for using AI to facilitate molecular-probe design, which is still in its early stages. Deep-learning models have demonstrated increased efficiency and image quality for PET reconstruction from sinogram data. Generative adversarial networks (GANs), which are paired neural networks that are jointly trained to generate and classify images, have found applications in modality transformation, artifact reduction, and synthetic-PET-image generation. Some AI applications, based either partly or completely on neural-network approaches, have demonstrated superior differential-diagnosis generation relative to radiologists. However, AI models have a history of brittleness, and physicians and patients may not trust AI applications that cannot explain their reasoning. To date, the majority of molecular-imaging applications of AI have been confined to research projects, and are only beginning to find their ways into routine clinical workflows via commercialization and, in some cases, integration into scanner hardware. Evaluation of actual clinical products will yield more realistic assessments of AI's utility in molecular imaging.


Phenolic content and antioxidant properties of soybean (Glycine max (L.) Merr.) seeds.
The contents and antioxidant ability of various classes of phenolic compounds present in the seeds of twenty soybean hybrids were evaluated. Total phenolics, tannins and proanthocyanidins were determined spectrophotometrically, after extraction of seeds with 70% aqueous acetone. In addition, the flavonoid contents were determined. The antioxidant activity of aqueous acetone extracts was evaluated by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity assay. The highest contents of total phenolics were found in Serbian cultivar 1511 and Chinese cultivar LN92-7369, which also displayed the highest total antioxidant activity. Conversely, genotypes poor in phenolics also showed low levels of DPPH-radical scavenging activity. The results suggested that besides protein and oil contents, the phenolic contents should be also considered as an important characteristic feature of soybean seeds, and as a potential selection criterion for antioxidant activity in soybean.


Cut training costs with computer-based tutorials.
New clinical information systems use client-server software applications that require knowledge of personal computers, Microsoft Windows, and mouse control. Here, the authors describe a low-cost training method to supplement classroom teaching.


Standardizing Screw Head Orthopaedic Trauma Implants. Is There an Interest? An OTA Member Survey.
To determine the level of interest in standardization of design for fracture fixation implants within the orthopaedic trauma community. A survey regarding implant removal concerns and implant design preferences was electronically distributed to members of Orthopaedic Trauma Association (OTA). Three hundred thirty respondents participated in the survey (response rate of 19%) Only 2.4% of respondents indicated a consistent ability to identify the implant vendor and/or manufacturer of retained implants in the preoperative planning phase of care; >75% of respondents reported investing a minimum of an additional 30 minutes to their operative times to remove screw(s) to successfully extract a plate. More than 80% of respondents reported multiple occurrences requiring modification of their surgical plans because of implants that could not be removed, preventing the completion of the planned procedure. The majority of respondents expressed interest in the adoption of standardized locking head screws for mini, small, and large screw sizes sets (63.5%); 84.8% of respondents desire standardization of screw head and driver sizes across the industry. Also, 83.6% of respondents expressed interest in the adoption of a standardized locking screw head for each cannulated screw, regardless of manufacturer. Members of the orthopaedic community, as represented in the membership of the OTA, are interested in the adoption of universal standards in implant design. Establishing standard screw heads and drivers will optimize the delivery of operative care.


Diagnosis and treatment of ovarian cysts in children.
Seventeen girls were treated following the diagnosis of ovarian cysts. Four patients were operated on within the first 6 months of age and the other 13 patients were 10-15 years old at the time of diagnosis. Antenatal diagnosis was made in 3 cases. The presenting symptoms in the infants were distended abdomen in 3 cases, abdominal pain in 1 and vomiting in 1. In the older children the presenting symptoms were abdominal pain in 12, vomiting in 5 and elevated temperature in 6. Preoperative ultrasound was performed in 5 patients, 3 neonates and 2 older children. Sixteen of the 17 girls were operated on. The indication for surgery was an ovarian cyst with complication in the infants and in the older children the suspicion of acute appendicitis. The operative procedure was cyst uncapping in 7 cases, salpingo-oophorectomy in 4, ovarian resection in 2, ovarian fixation only in 1 and no ovarian intervention in 2. Asymptomatic infants with an ovarian cyst less than 4 cm in diameter can be managed conservatively. Surgery can be recommended after documented change of the cyst on ultrasound, large cysts giving rise to symptoms or presentation of an acute abdomen. Salvage of variable ovarian tissue is desirable.


Structural reorganization of SHP2 by oncogenic mutations and implications for oncoprotein resistance to allosteric inhibition.
Activating mutations in PTPN11, encoding the cytosolic protein tyrosine phosphatase SHP2, result in developmental disorders and act as oncogenic drivers in patients with hematologic cancers. The allosteric inhibitor SHP099 stabilizes the wild-type SHP2 enzyme in an autoinhibited conformation that is itself destabilized by oncogenic mutations. Here, we report the impact of the highly activated and most frequently observed mutation, E76K, on the structure of SHP2, and investigate the effect of E76K and other oncogenic mutations on allosteric inhibition by SHP099. SHP2E76K adopts an open conformation but can be restored to the closed, autoinhibited conformation, near-identical to the unoccupied wild-type enzyme, when complexed with SHP099. SHP099 inhibitory activity against oncogenic SHP2 variants in vitro and in cells scales inversely with the activating strength of the mutation, indicating that either oncoselective or vastly more potent inhibitors will be necessary to suppress oncogenic signaling by the most strongly activating SHP2 mutations in cancer.


Nuclear RNA polymerase activity in tumor-host livers.
This study has evaluated changes in RNA synthesis in livers under the distant influence of a malignant tumor. A transplantable-induced sarcoma (MCG 101), transplanted on inbred adult mice (C57BL/6J), was used. Activities of DNA-dependent RNA polymerase (EC 184.108.40.206) were measured in relation to RNA content and translational activity. Liver nuclei from freely fed sarcoma-bearing mice had increased RNA synthesis. As a consequence of this, RNA content per DNA was increased in liver tissue. This was independent of depressed food intake and malnutrition. Elevated RNA synthesis, proportional to the tumor burden was due to an increased proportion of chromatin-engaged RNA polymerase I and II activities. RNA polymerase III activity (template-engaged form) was unchanged when evaluated in isolated nuclei, but appeared to be increased in partially purified extracts of nuclei. RNA content in tumor-host liver was a composite of increased levels of rRNA and tRNA, whereas the levels of poly(A)+ mRNA could not be measured as increased. Overall translational activities in vitro of mRNA from liver tissue of tumor-bearing, pair-weighed, and freely fed tumor-free controls were qualitatively and quantitatively different. mRNA from tumor-bearing mice directed an increased synthesis, particularly of larger proteins (above 55,000 daltons) compared with control animals. The results support the conclusion that previous evidence of elevated net protein synthesis in tumor-host liver is accompanied by increased transcription of genes coding for RNA and also for some or several hepatic proteins.


Roles of v-erbA homodimers and heterodimers in mediating dominant negative activity by v-erbA.
v-erbA, a viral oncogenic homolog of thyroid hormone receptor (TR), blocks the effect of T3 in TR-mediated transcription. The mechanism(s) for this dominant negative effect by v-erbA on TRs is unknown but may involve competition between v-erbA and TR-containing complexes for binding to thyroid hormone response elements (TREs) and/or protein-protein interactions between v-erbA and TR. To investigate these potential mechanisms, we used the electrophoretic mobility shift assay to compare in vitro translated v-erbA and TR alpha binding to two TREs-chick lysozyme TRE (F2) and direct repeat TRE (DR4). v-erbA bound as a homodimer to these TREs, whereas TR alpha bound as a homodimer and monomer. T3 decreased TR alpha homodimer binding to the TREs as we reported previously; however, surprisingly, high concentrations of T3 (10(-6) M) also decreased v-erbA homodimer binding to the TREs. Additionally, v-erbA formed heterodimers with nuclear proteins such as retinoid X receptor and T3 receptor auxiliary protein as well as with TR alpha. These dimers remained bound to DNA in the presence of T3. Finally, v-erbA could not mediate ligand-dependent transcriptional activation even at 10(-6) M T3 but could block ligand-dependent TR-mediated transactivation in co-transfection experiments. v-erbA also exhibited differential dominant negative activity on F2 and DR4 suggesting that half-site sequence and/or orientation may influence v-erbA-dominant negative activity. In sum, there are multiple v-erbA complexes that bind to TREs in the presence of T3, which all may contribute to v-erbA's dominant negative effect on TR-mediated transcription by competing with TR-containing complexes for binding to TREs.


Authentication of commercial spices based on the similarities between gas chromatographic fingerprints.
A simple and solvent-free method was developed for the authentication of commercial spices. The similarities between gas chromatographic fingerprints were measured using similarity indices and multivariate data analyses, as morphological differentiation between dried powders and small spice particles was challenging. The volatile compounds present in 11 spices (i.e. allspice, anise, black pepper, caraway, clove, coriander, cumin, dill, fennel, star anise, and white pepper) were extracted by headspace solid-phase microextraction, and analysed by gas chromatography-mass spectrometry. The largest 10 peaks were selected from each total ion chromatogram, and a total of 65 volatiles were tentatively identified. The similarity indices (i.e. the congruence coefficients) were calculated using the data matrices of the identified compound relative peak areas to differentiate between two sets of fingerprints. Where pairs of similar fingerprints produced high congruence coefficients (>0.80), distinctive volatile markers were employed to distinguish between these samples. In addition, hierarchical cluster analysis and principal component analysis were performed to visualise the similarity among fingerprints, and the analysed spices were grouped and characterised according to their distinctive major components. This method is suitable for screening unknown spices, and can therefore be employed to evaluate the quality and authenticity of various spices. © 2017 Society of Chemical Industry.


Transient inhibition of cell proliferation does not compromise self-renewal of mouse embryonic stem cells.
Embryonic stem cells (ESCs) have unlimited capacity for self-renewal and can differentiate into various cell types when induced. They also have an unusual cell cycle control mechanism driven by constitutively active cyclin dependent kinases (Cdks). In mouse ESCs (mESCs). It is proposed that the rapid cell proliferation could be a necessary part of mechanisms that maintain mESC self-renewal and pluripotency, but this hypothesis is not in line with the finding in human ESCs (hESCs) that the length of the cell cycle is similar to differentiated cells. Therefore, whether rapid cell proliferation is essential for the maintenance of mESC state remains unclear. We provide insight into this uncertainty through chemical intervention of mESC cell cycle. We report here that inhibition of Cdks with olomoucine II can dramatically slow down cell proliferation of mESCs with concurrent down-regulation of cyclin A, B and E, and the activation of the Rb pathway. However, mESCs display can recover upon the removal of olomoucine II and are able to resume normal cell proliferation without losing self-renewal and pluripotency, as demonstrated by the expression of ESC markers, colony formation, embryoid body formation, and induced differentiation. We provide a mechanistic explanation for these observations by demonstrating that Oct4 and Nanog, two major transcription factors that play critical roles in the maintenance of ESC properties, are up-regulated via de novo protein synthesis when the cells are exposed to olomoucine II. Together, our data suggest that short-term inhibition of cell proliferation does not compromise the basic properties of mESCs.


Molecular characterization and prevalence of Halarachne halichoeri in threatened southern sea otters (Enhydra lutris nereis).
Parasitism, particularly in concert with other sublethal stressors, may play an important, yet underappreciated role in morbidity and mortality of threatened species. During necropsy of southern sea otters (Enhydra lutra nereis) from California submitted to the Marine Wildlife Veterinary Care and Research Center's Sea Otter Necropsy Program between 1999 and 2017, pathologists occasionally observed nasopulmonary mites infesting the respiratory tracts. Infestation was sometimes accompanied by lesions reflective of mite-associated host tissue damage and respiratory illness. Our objectives were to estimate prevalence of nasopulmonary mites, determine the taxonomic identity of the observed mites, and create a DNA reference for these organisms in southern sea otters as an aid in population management. Using unique morphological characteristics discerned via light and scanning electron microscopy (SEM), we identified the mites as Halarachne halichoeri, a species typically associated with harbor seals (Phoca vitiluna). The 18S, 16S, 28S and ITS1-2 genetic regions were sequenced and submitted to GenBank. We observed H. halichoeri mites in 25.6% (95% CI 19.9-33.4%). of southern sea otters from a subset of necropsies performed between 2012 and 2017. This is the first documentation of H. halichoeri in southern sea otters and is suggestive of parasite exchange between sea otters and harbor seals.


Mapping the functional domains of the beta-adrenergic receptor.
The beta-adrenergic receptor (beta AR) serves as a model system for analysis of the structure-function relationships of G-protein-coupled receptors. Genetic analysis of the beta AR has demonstrated that the ligand-binding domain of this protein lies within the hydrophobic putative transmembrane core, involving specific amino acid residues in several of the transmembrane helices of the receptor. Site-directed mutagenesis of the receptor in conjunction with structural alterations of the ligands has revealed specific molecular interactions that are important for recognition of the ligand by the receptor. In addition, cytoplasmically exposed regions of the beta AR that are required for the activation of Gs have been identified. Because of the structural similarities among G-protein-coupled receptors, information gained from genetic analysis of the beta AR should prove useful in the development of specific agonists and antagonists for other receptors of this class.


Tuberculosis in children treated with second-line drugs under programmatic conditions in Lima, Peru.
To characterise childhood tuberculosis (TB) treated with second-line drugs (SLDs) in Lima, Peru. Results for the age groups <5 and 5-14 years were compared and treatment outcomes were assessed in cases reported between 2011 and 2015 from six districts of Lima. Of 96 reported cases, 82 were evaluated. Among these, 59% were boys; the median age was 8 years and 32% were aged <5 years. Contact with a TB case was reported in 82% of cases; 90% were treatment-naïve, 98% had pulmonary localisation and 50% underwent the tuberculin skin test (purified protein derivative), with induration 10 mm in 88%. A positive smear was found in 40%, all in the 5-14 years age group, and 46% were culture-positive. Only 26% had confirmed multidrug-resistant TB, 90% of whom were in the 5-14 years age group. SLDs for confirmed or probable drug-resistant TB (DR-TB) were administered to all cases, with a high proportion of success (over 83%), no failures or deaths and a high proportion of loss to follow-up. The main indication for SLDs in childhood TB was the empirical treatment of DR-TB due to contact with one or more identified DR-TB patients. Bacteriological confirmation was limited; however, treatment success was adequate.


Variability in Toxicity of Plastic Leachates as a Function of Weathering and Polymer Type: A Screening Study with the Copepod Nitocra spinipes.
AbstractThe production and use of plastic over many decades has resulted in its accumulation in the world's oceans. Plastic debris poses a range of potential risks to the marine environment and its biota. Especially, the potential hazards of small plastic debris and chemicals associated with plastic have not been extensively studied. When buoyant plastic is exposed to ultraviolet radiation, it will slowly degrade and leach chemicals into surrounding waters. These leachates can include additives, sorbed organic pollutants, and degradation products of the plastic polymers. While most hazard assessments have focused on studying adverse effects due to the uptake of plastic, toxicity studies of the leachates of plastics are less common. To begin to address this knowledge gap, we studied the acute toxicity of leachates from diverse plastics in the harpacticoid copepod Nitocra spinipes. Our results show that leachates caused a higher toxicity after plastic was exposed to ultraviolet light compared to leaching in darkness. We observed differences in toxicity for different polymer types: polyvinyl chloride and polypropylene resulted in the most toxic leachates, while polystyrene and poly[ethylene terephthalate] were least toxic. Furthermore, we observed increased toxicity of leachates from some plastics that had been weathered in the real marine environment compared to matching new materials. Our results indicate that both weathering condition and polymer type influence the toxicity of plastic leachates.


The autogenous dermis-fat orbital implant in children.
The autogenous dermis-fat graft orbital implant is one alternative for volume augmentation in the adult anophthalmic socket. We have reviewed our experience with dermis-fat grafts after primary enucleation in the pediatric population. We reviewed the clinic charts of all patients under 8 years of age who underwent primary enucleation combined with dermis-fat graft implantation. We assessed subsequent orbital growth by measuring lid and fissure symmetry, superficial soft tissue socket volume, prosthetic fit, and periorbital symmetry. All measurements were obtained by 2 of the authors. Eight children had primary enucleations combined with a dermis-fat implant (Table 1). Age at the time of surgery ranged from 1 week to 8 years. Indications for enucleation were retinoblastoma (4), malignant teratoid medulloepithelioma (1), blind, painful eye secondary to trauma (2), or phthisis secondary to end-stage retinopathy of prematurity (1). Two patients were lost to follow up at 3 months and 6 months post enucleation. The remaining 6 patients had between 1 year and 6 years of follow up. All implants maintained appropriate volume allowing proper prosthetic fit and facial symmetry. Two children experienced excessive growth of their implants, managed by surgical debulking. Two children developed a central graft ulcer, managed by superficial revision and reclosure. We have observed that dermis-fat grafts in pediatric primary enucleations demonstrate good maintenance of orbital soft tissue volume and periorbital symmetry. The complications of excessive dermis-fat growth or central ulceration were easily corrected. We feel this is a promising implant for the pediatric anophthalmic socket.


Mapping the protein profile involved in the biotransformation of organoarsenicals using an arsenic metabolizing bacterium.
Alkaliphilus oremlandii strain OhILAs, a gram-positive bacterium, has been shown to ferment lactate as well as use arsenate and roxarsone as a terminal electron acceptor. This study examines the proteome expressed under four growth conditions to further elucidate the bacterial metabolism of inorganic and organic arsenic. The four growth conditions include, sodium lactate (as fermentative control), sodium lactate with 3-nitro-4-hydroxybenzenearsonic acid (roxarsone), sodium lactate with 3-amino-4-hydroxybenzenearsonic acid (3A4HBAA), and sodium lactate with sodium arsenate. Shotgun proteomics using LC-MS/MS was performed on the soluble cytoplasm as well as solubilized membrane proteins using perfluorooctanoic acid, a surfactant with properties similar to sodium dodecyl sulfate. The MS/MS data were analyzed using the Spectrum Mills Proteomic Workbench. Positive protein matches were confirmed with protein scores of 20 or greater and the presence of two or more peptides among the three technical replicates. A total of 1357 proteins (out of 2836 predicted) were identified with 791 in sodium lactate, 816 in sodium lactate and roxarsone, 715 in sodium lactate and 3A4HBAA, and 733 in sodium lactate and arsenate. The relative abundance of each protein was determined using a method called normalized spectral abundance factor (NSAF). Proteins that were identified in both the control and the experimental conditions were compared using the Power Law Global Error Model (PLGEM) to determine proteins that were significantly up or down regulated. All putative proteins were assigned functions and pathways using the COG databases. However, a large number of proteins were classified as hypothetical or had unknown function. Using the statistical information and known functionalities of the identified proteins, a pathway for the degradation of roxarsone and 3A4HBAA by A. oremlandii strain OhILAs is proposed.


Meal-stimulated absorption of water and electrolytes in canine jejunum.
After a meal, the absorption of water and electrolytes from the jejunal lumen increases. This meal-induced jejunal absorption occurs in jejunal segments out of normal gastrointestinal continuity. The experimental model used 25-cm proximal jejunal Thiry-Vella loops in awake dogs (n = 72 observations) to evaluate the mechanisms involved in meal-induced jejunal absorption, seeking to define the source or sources of the proabsorptive signal. Specifically, we evaluated the jejunal absorptive response to a standard meal, a standard meal plus cholinergic blockage using atropine, a sham-fed meal, a gavage-fed meal, and gastric distension with balloon and gavage water. Both the standard meal and the gavage-fed meal induced a prompt, sustained, and significant (P less than 0.0001) increase in the absorption of H2O, Na+, and Cl-. Atropine significantly reduced the magnitude of the postmeal absorptive response (P less than 0.05) compared with the standard meal alone. The sham-fed meal, gastric balloon distension, and gavage water did not alter jejunal absorption. Vagal nerve integrity after cervical esophageal manipulation was verified by gastric acid output and gastrin response to stimuli. These data support a role for cholinergic modulation of meal-stimulated jejunal absorption via a cephalic-phase-independent and gastric-distension-independent mechanism.


Chemoselective efficient synthesis of functionalized β-oxonitriles through cyanomethylation of Weinreb amides.
A synthesis of β-oxonitriles is reported via the generation of R(1)R(2)CLiCN species followed by the trapping with variously decorated Weinreb amides. The optimization study revealed that lithiation of acetonitriles is best accomplished by deprotonation with MeLi-LiBr at low temperature. The protocol can be conveniently adapted to the synthesis of α-mono or α,α-disubstituted cyanoketones. (15)N- and (17)O-NMR data are reported for selected compounds.


Health and the Economy in the United States, from 1750 to the Present.
I discuss the health transition in the United States, bringing new data to bear on health indicators, and investigating the changing relationship between health, income, and the environment. I argue that scientific advances played an outsize role and that health improvements were largest among the poor. Health improvements were not a precondition for modern economic growth. The gains to health are largest when the economy has moved from "brawn" to "brains" because this is when the wage returns to education are high, leading the healthy to obtain more education. More education may improve use of health knowledge, producing a virtuous cycle.


Copper induces cell death by targeting lipoylated TCA cycle proteins.
Copper is an essential cofactor for all organisms, and yet it becomes toxic if concentrations exceed a threshold maintained by evolutionarily conserved homeostatic mechanisms. How excess copper induces cell death, however, is unknown. Here, we show in human cells that copper-dependent, regulated cell death is distinct from known death mechanisms and is dependent on mitochondrial respiration. We show that copper-dependent death occurs by means of direct binding of copper to lipoylated components of the tricarboxylic acid (TCA) cycle. This results in lipoylated protein aggregation and subsequent iron-sulfur cluster protein loss, which leads to proteotoxic stress and ultimately cell death. These findings may explain the need for ancient copper homeostatic mechanisms.


Neuroprotective effects of catechins in an experimental Parkinson's disease model and SK-N-AS cells: evaluation of cell viability, anti-inflammatory and anti-apoptotic effects.
The aim of the study was to establish an in vitro Parkinson's disease (PD) model and to investigate the cell viability, anti-inflammatory, anti-apoptotic and neuroprotective effects of catechin and EGCG in SK-N-AS and in vitro PD model cells. SK-N-AS human neuroblastoma cells were used. To develop an in vitro PD model, SK-N-AS cells were exposed to 6-hydroxydopamine. Model control was performed after ELISA analysis of dopamine and α-synuclein levels in the culture medium. Catechin and EGCG were administered to SK-N-AS and in vitro PD model cells. Cell viability was measured using MTT assay and trypan blue staining. Anti-inflammatory and anti-apoptotic activities of catechin and EGCG were investigated by indirect immunocytochemistry using anti-TNF-α, anti-IL-1β and anti-caspase-3. After 24 hours of 6-hydroxydopamine administration at 50 μM, higher αlfa-synuclein and lower dopamine levels were found in PD model than SK-N-AS cells. Cell viability was similar between SK-N-AS and in vitro PD model cells. Treatment with both bioactive components increased cell viability of in vitro PD model cells. Caspase-3 immunoreactivity was significantly reduced in SK-N-AS and PD model cells after EGCG administration, while it was decreased only in PD model cells after catechin administration. IL-1β staining intensity weakened after catechin administration in PD model cells, after EGCG administration in SK-N-AS cells. TNF-α staining intensity was similar in both cells. Catechin and EGCG increased cell viability in PD model neuron cells. Both components showed anti-apoptotic and anti-inflammatory effects. Catechin may be more effective in preventing damage to neurons PD.


Analysis of T-cell receptor gene rearrangement and expression in human natural killer clones.
A series of clones of human natural killer (NK) cells was characterized with respect to expression of the Ti alpha and Ti beta genes of the T-cell receptor. T11+T3+ NK clones contained Ti alpha and Ti beta RNA transcripts and expressed disulfide-linked heterodimers, demonstrating the presence of a functional T-cell receptor. In contrast, T11+T3- NK clones expressed only 1.0-kilobase truncated Ti beta transcripts, without a Ti alpha transcript and no detectable surface Ti protein. Since previous studies demonstrated that Ti beta gene activation precedes Ti alpha gene activation in thymic ontogeny, the T11+T3- NK cells appear to be derived from T-lineage precursors.


Multiple sclerosis: cerebral circulation time.
To assess cerebral circulation times (CCTs) in patients with multiple sclerosis (MS) and control subjects by using contrast material-enhanced ultrasonography (US) to determine whether vascular abnormalities can be detected in this disease. This study was approved by the local ethics committee, and informed consent was obtained from all subjects. One hundred three patients with MS and 42 control subjects underwent extracranial and transcranial venous echo-color Doppler ultrasonography (US) and contrast-enhanced US. CCT was defined as the difference in arrival time of the US contrast agent bolus between the carotid artery and the internal jugular vein. The presence of chronic cerebrospinal venous insufficiency (CCSVI) was defined according to previously reported criteria for the extracranial and transcranial US techniques. Nonparametric statistics, including the Mann-Whitney U test and the Kruskal-Wallis analysis of variance, were used to compare contrast-enhanced US parameters between groups. The longest and average CCTs were substantially prolonged in patients with MS compared with those in control subjects (median longest CCT in patients with MS, 6.47 seconds [range, 3.29-29.24 seconds]; that in control subjects, 5.54 seconds [range, 2.57-7.63 seconds]; P < .001; median average CCT in patients with MS, 5.76 seconds [range, 2.64-17.51 seconds]; that in control subjects, 5.01 seconds [range, 2.57-7.06 seconds]; P < .002). No correlation was found between CCTs and clinical parameters. The prevalence of CCSVI was higher in patients with MS than in control subjects (77% vs 28%, P < .0001). CCT was not significantly different between patients with MS who had CCSVI and patients with MS who did not (P = .182). These results suggest that contrast-enhanced US with CCT assessment may have a role in the evaluation of cerebral blood flow in patients with MS and that a vascular impairment could be associated with MS. The finding of a prolonged CCT at contrast-enhanced US does not result from outflow impairment. Further studies are required to verify these observations and to clarify if CCT and CCSVI have any physiologic and clinical relevance in MS.


Impaired auditory temporal selectivity in the inferior colliculus of aged Mongolian gerbils.
Aged humans show severe difficulties in temporal auditory processing tasks (e.g., speech recognition in noise, low-frequency sound localization, gap detection). A degradation of auditory function with age is also evident in experimental animals. To investigate age-related changes in temporal processing, we compared extracellular responses to temporally variable pulse trains and human speech in the inferior colliculus of young adult (3 month) and aged (3 years) Mongolian gerbils. We observed a significant decrease of selectivity to the pulse trains in neuronal responses from aged animals. This decrease in selectivity led, on the population level, to an increase in signal correlations and therefore a decrease in heterogeneity of temporal receptive fields and a decreased efficiency in encoding of speech signals. A decrease in selectivity to temporal modulations is consistent with a downregulation of the inhibitory transmitter system in aged animals. These alterations in temporal processing could underlie declines in the aging auditory system, which are unrelated to peripheral hearing loss. These declines cannot be compensated by traditional hearing aids (that rely on amplification of sound) but may rather require pharmacological treatment.


Cetia pacifica gen. nov., sp. nov., a chemolithoautotrophic, thermophilic, nitrate-ammonifying bacterium from a deep-sea hydrothermal vent.
A thermophilic, anaerobic, chemolithoautotrophic bacterium, strain TB-6(T), was isolated from a deep-sea hydrothermal vent located on the East Pacific Rise at 9° N. The cells were Gram-staining-negative and rod-shaped with one or more polar flagella. Cell size was approximately 1-1.5 µm in length and 0.5 µm in width. Strain TB-6(T) grew between 45 and 70 °C (optimum 55-60 °C), 0 and 35 g NaCl l(-1) (optimum 20-30 g l(-1)) and pH 4.5 and 7.5 (optimum pH 5.5-6.0). Generation time under optimal conditions was 2 h. Growth of strain TB-6(T) occurred with H2 as the energy source, CO2 as the carbon source and nitrate or sulfur as electron acceptors, with formation of ammonium or hydrogen sulfide, respectively. Acetate, (+)-d-glucose, Casamino acids, sucrose and yeast extract were not used as carbon and energy sources. Inhibition of growth occurred in the presence of lactate, peptone and tryptone under a H2/CO2 (80 : 20; 200 kPa) gas phase. Thiosulfate, sulfite, arsenate, selenate and oxygen were not used as electron acceptors. The G+C content of the genomic DNA was 36.8 mol%. Phylogenetic analysis of the 16S rRNA gene of strain TB-6(T) showed that this organism branched separately from the three most closely related genera, Caminibacter , Nautilia and Lebetimonas , within the family Nautiliaceae . Strain TB-6(T) contained several unique fatty acids in comparison with other members of the family Nautiliaceae . Based on experimental evidence, it is proposed that the organism represents a novel species and genus within the family Nautiliaceae , Cetia pacifica, gen. nov., sp. nov. The type strain is TB-6(T) ( = DSM 27783(T) = JCM 19563(T)).


Protective effect of cilazapril on the cerebral circulation.
The goal of an antihypertensive treatment is to prevent "end-organ" damage. Cerebral vascular complications are among the most important because they are life threatening and can occur even at an early stage of the disease. Recently, it has been shown that cilazapril can decrease the mortality of stroke-prone rats, suggesting a decrease in the incidence of strokes, which occur spontaneously in these animals. The present article reviews the different functional and morphological changes that may explain the cerebral protective effects of cilazapril, such as the normalization of cerebral vascular reserve, decrease in the media, increase in the external diameter, and normalization of the mechanics and endothelial function of cerebral arterioles. In addition, the inhibition by cilazapril of injury-induced proliferation of smooth muscle cells and the infiltration of the endothelium by macrophages could prevent the development of atherosclerosis.


Cell cycle-dependent expression of Kv3.4 channels modulates proliferation of human uterine artery smooth muscle cells.
Vascular smooth muscle cell (VSMC) proliferation is involved in cardiovascular pathologies associated with unwanted arterial wall remodelling. Coordinated changes in the expression of several K+ channels have been found to be important elements in the phenotypic switch of VSMCs towards proliferation. We have previously demonstrated the association of functional expression of Kv3.4 channels with proliferation of human uterine VSMCs. Here, we sought to gain deeper insight on the relationship between Kv3.4 channels and cell cycle progression in this preparation. Expression and function of Kv3.4 channels along the cell cycle was explored in uterine VSMCs synchronized at different checkpoints, combining real-time PCR, western blotting, and electrophysiological techniques. Flow cytometry, Ki67 expression and BrdU incorporation techniques allowed us to explore the effects of Kv3.4 channels blockade on cell cycle distribution. We found cyclic changes in Kv3.4 and MiRP2 mRNA and protein expression along the cell cycle. Functional studies showed that Kv3.4 current amplitude and Kv3.4 channels contribution to cell excitability increased in proliferating cells. Finally, both Kv3.4 blockers and Kv3.4 knockdown with siRNA reduced the proportion of proliferating VSMCs. Our data indicate that Kv3.4 channels exert a permissive role in the cell cycle progression of proliferating uterine VSMCs, as their blockade induces cell cycle arrest after G2/M phase completion. The modulation of resting membrane potential (V(M)) by Kv3.4 channels in proliferating VSMCs suggests that their role in cell cycle progression could be at least in part mediated by their contribution to the hyperpolarizing signal needed to progress through the G1 phase.


Concordance of coronary artery calcium estimates between MDCT and electron beam tomography.
The objective of our study was to compare MDCT with electron beam tomography (EBT) for the quantification of coronary artery calcification (CAC). Sixty-eight patients underwent both MDCT and EBT within 2 months for the quantification of CAC. The images were scored in a blinded fashion and independently by two observers with a minimum of 7 days between the interpretations of images obtained from one scanner type to the other. Presence versus absence of CAC was discordant by EBT versus MDCT in 6% (n = 4) of the cases by observer 1, with one of these cases also discordant by observer 2. All cases except one (aortic calcium misidentified as CAC) were among those with a mean Agatston score of less than 5 present on EBT but absent on MDCT. EBT and MDCT scores correlated well (r = 0.98-0.99). The relative median variability between EBT and MDCT for the Agatston score was 24% for observer 1 and 27% for observer 2 and was 18% and 14%, respectively, for volume score (average for both observers: 27% for Agatston score and 16% for volume score). Scores were higher for EBT than MDCT in approximately half of the cases, with little systematic difference between the two (median EBT-MDCT difference: Agatston score, -0.55; volume score, 3.4 mm3). The absolute median difference averaged for the two observers was 28.75 for the Agatston score and 15.4 mm3 for the volume score. Differences in CAC measurements using EBT and MDCT are similar to interscan differences in CAC measurements previously reported for EBT or for other MDCT scanners individually.


Relationships between minute characteristics of finger ridges and pattern size and shape.
We present results of a multivariate analysis of several characteristics of the ridge pattern on the tip of fingers III and IV of the right hand, with special consideration of the minute properties of ridges. The analysis is based on a sample of 38 females and 63 males of European origin. Results of factor analysis for the male sample reveal that pattern type and size load on one factor. Furthermore, there are separate factors for the two types of minutiae . If characters on both fingers are considered jointly, there are seven factors, identical for both fingers on some variables, but unique for minutiae , especially for junctions, on each finger. These results are consistent with evidence obtained in our previous study that junctions and pattern type are largely independent correlates with tactile sensitivity.


Prolonged Thrombocytopenia in a Case of MIS-C in a Vaccinated Child.
Multisystem inflammatory syndrome in children (MIS-C) has been extensively described in patients following severe acute respiratory syndrome coronavirus 2 infection. There are now questions about what MIS-C may look like in vaccinated children. Multisystem inflammatory syndrome in children has many clinical and laboratory features in common with other inflammatory disorders including Kawasaki disease and toxic shock syndrome. Rheumatologic conditions can present with similar musculoskeletal complaints and elevated inflammatory markers. Laboratory markers and clinical symptoms of MIS-C usually improve once therapy is begun. We describe a child with persistent thrombocytopenia as an example of variable presentation of MIS-C in vaccinated children. This case report discusses an atypical progression of MIS-C in a vaccinated child with a known prior positive COVID-19 polymerase chain reaction (PCR) test. She presented with nonspecific abdominal pain and fever and was found to have elevated inflammatory markers, lymphopenia, and thrombocytopenia. Intravenous immunoglobulin and steroid treatment failed to induce rapid recovery in her clinical condition or thrombocytopenia. Rheumatologic, hematologic, oncologic, and infectious causes were considered and worked up due to the uncertainty of her case and persistence of pancytopenia but ultimately were ruled out with extensive testing and monitoring. It was key to include a broad differential including viral-induced bone marrow suppression, idiopathic thrombocytopenic purpura, secondary hemophagocytic lymphohistiocytosis, systemic juvenile idiopathic arthritis, and malignancy. The spectrum of MIS-C and response to treatment continues to evolve, and prior vaccination in this child's case complicated the clinical picture further. Additional evaluation of MIS-C in vaccinated cases will permit characterization of the range of MIS-C presentation and response to standard therapy.


Obesity management in gynecologic cancer survivors: provider practices and attitudes.
Obesity is associated with the development and risk of death from several women's cancers. The study objective was to describe and compare oncologic providers' attitudes and practices as they relate to obesity counseling and management in cancer survivors. Society of Gynecologic Oncology members (n = 924) were surveyed with the use of a web-based, electronic questionnaire. χ(2) and Fisher exact tests were used to analyze responses. Of the 240 respondents (30%), 92.9% were practicing gynecologic oncologists or fellows, and 5.1% were allied health professionals. Median age was 42 years; 50.8% of the respondents were female. Of the respondents, 42.7% reported that they themselves were overweight/obese and that ≥50% of their survivor patients were overweight/obese. Additionaly, 82% of the respondents believed that discussing weight would not harm the doctor-patient relationship. Most of the respondents (95%) agreed that addressing lifestyle modifications with survivors is important. Respondents believed that gynecologic oncologists (85.1%) and primary care providers (84.5%) were responsible for addressing obesity. More providers who were ≤42 years old reported undergoing obesity management training (P < .001) and were more likely to believe that survivors would benefit from obesity education than providers who were >42 years old (P = .017). After initial counseling, 81.5% of the respondents referred survivors to other providers for obesity interventions. Oncology provider respondents believe that addressing obesity with cancer survivors is important. Providers believed themselves to be responsible for initial counseling but believed that obesity interventions should be directed by other specialists. Further research is needed to identify barriers to care for obese cancer survivors and to improve physician engagement with obesity counseling in the "teachable moment" that is provided by a new cancer diagnosis.


Two-dimensional identification and localization of isomers in crystallin peptides using TWIM-MS.
Recent studies have illuminated connections between spontaneous chemical reactions that cause isomerization at specific protein residues and various age-related diseases including cataracts and Alzheimer's. These discoveries provide impetus for better analytical methods to detect and characterize isomerization in proteins, which will enable a more complete understanding of the underlying relationship between these modifications and biology. Herein we employ a two-dimensional approach for identification of peptides isomers that also includes pinpointing of the modified residue. Collision-induced dissociation is used to fragment ions in the first dimension, followed by separation of the fragments with travelling-wave ion mobility. By comparing data obtained from both isomers, differences in either fragment-ion intensities or arrival-time distributions can be used to identify isomeric forms and the specific site of modification within the peptides. Synthetic peptide standards with sequences derived from long-lived proteins in the eye lens and isomerization at serine, aspartic acid, and glutamic acid were examined. Although both dimensions are capable of isomer identification, ion mobility is much better at determining the site of modification. In general, separation of isomeric forms by ion mobility is possible but does not follow predictable trends dictated by sequence or fragment-ion length. In most cases, however, the site of isomerization can be precisely determined.


Monkeypox: another pandemic in the making?
Monkeypox is a zoonotic disease caused by the monkeypox virus, which is a member of the Poxviridae family of viruses. It is transmitted through direct or indirect contact with fluid secretions. Initial symptoms include fever, chills, headache, and malaise, followed by a maculopapular rash that starts on the face and progresses centrifugally. Polymerase chain reaction is the preferred laboratory test for the diagnosis, and management is mostly supportive. The clinical presentation of monkeypox is quite similar to that of another member of the Poxviridae family: smallpox, which wreaked havoc in the 20th century, before being eradicated with the help of the vaccinia virus vaccine in 1977. This vaccine protects not only against smallpox but also monkeypox; therefore, when use of this vaccine was discontinued, monkeypox had a new susceptible population to infect and way to proliferate and evolve. Initially the disease spread in Africa, but now the more evolved monkeypox is quickly spreading to other countries. On July 23, 2022, the World Health Organization declared this multicountry outbreak a public health emergency of international concern. Given its mutating ability and high transmissibility, we need to quickly devise measures to control this virus before it turns into a pandemic.


Genome-based analysis of Chlamydomonas reinhardtii exoribonucleases and poly(A) polymerases predicts unexpected organellar and exosomal features.
Enzymes from several gene families modify RNA molecules at their extremities. These reactions occur in several cellular compartments and affect every class of RNA. To assess the diversity of a subclass of these enzymes, we searched Chlamydomonas for open reading frames (ORFs) potentially encoding exoribonucleases, poly(A) polymerases, and proteins known to associate with and/or regulate them. The ORFs were further analyzed for indications of protein localization to the nucleus, cytosol, mitochondrion, and/or chloroplast. By comparing predicted proteins with homologs in Arabidopsis and yeast, we derived several tentative conclusions regarding RNA 5'- and 3'-end metabolism in Chlamydomonas. First, the alga possesses only one each of the following likely organellar enzymes: polynucleotide phosphorylase, hydrolytic exoribonuclease, poly(A) polymerase, and CCA transferase, a surprisingly small complement. Second, although the core of the nuclear/cytosolic exosome decay complex is well conserved, neither nucleus-specific activators nor the cytosolic exosome activators are present. Finally, our discovery of nine noncanonical poly(A) polymerases, a divergent family retaining the catalytic domains of conventional poly(A) polymerases, leads to the hypothesis that polyadenylation may play an especially important regulatory role throughout the Chlamydomonas cell, stabilizing some transcripts and targeting degradation machinery to others.


Virulence profile and genetic structure of a North Dakota population of Pyrenophora teres f. teres, the causal agent of net form net blotch of barley.
A Pyrenophora teres f. teres population in North Dakota was analyzed for virulence variation and genetic diversity using 75 monospore isolates that were collected across a 4-year period (2004 to 2007) from two North Dakota State University agricultural experiment stations at Fargo and Langdon. Pathogenicity tests by inoculation onto 22 barley differential lines at seedling stage revealed 49 pathotypes, indicating a wide range of pathogenic diversity. Two-way analysis of variance of disease ratings revealed a significant difference in the virulence among isolates and in the resistance among barley lines, as well as in the interactions between the two. 'CI5791', 'Algerian', and 'Heartland' were three barley lines showing a high level of seedling resistance to all North Dakota isolates tested; however, many previously reported resistance genes have been overcome. Forty multilocus genotypes were identified from this set of isolates by genotyping at 13 simple-sequence repeat loci. High percentages of clonal cultures were detected in the samplings from 2005 and 2007 in Fargo and 2005 in Langdon. Using a clone-corrected sample set, the mean gene diversity (h) was estimated to be 0.58, approximately the same for both locations. The calculated Wright's F(ST) value is small (0.11) but was significantly >0, indicating a significant differentiation between the Fargo and Langdon populations. In the gametic disequilibrium test, only 3 of 78 possible pairwise comparisons over all isolates showed significant (P < 0.05) nonrandom association, suggesting a random mating mode. Our results suggest that the populations from the two locations are derived from a common source and undergo frequent recombination. This research provides important information for barley breeders regarding development and deployment of cultivars with resistance to net form net blotch in this region.


Cell-free embryonic stem cell extract-mediated derivation of multipotent stem cells from NIH3T3 fibroblasts for functional and anatomical ischemic tissue repair.
The oocyte-independent source for the generation of pluripotent stem cells is among the ultimate goals in regenerative medicine. We report that on exposure to mouse embryonic stem cell (mESC) extracts, reversibly permeabilized NIH3T3 cells undergo dedifferentiation followed by stimulus-induced redifferentiation into multiple lineage cell types. Genome-wide expression profiling revealed significant differences between NIH3T3 control and ESC extract-treated NIH3T3 cells including the reactivation of ESC-specific transcripts. Epigenetically, ESC extracts induced CpG demethylation of Oct4 promoter, hyperacetylation of histones 3 and 4, and decreased lysine 9 (K-9) dimethylation of histone 3. In mouse models of surgically induced hindlimb ischemia or acute myocardial infarction transplantation of reprogrammed NIH3T3 cells significantly improved postinjury physiological functions and showed anatomic evidence of engraftment and transdifferentiation into skeletal muscle, endothelial cell, and cardiomyocytes. These data provide evidence for the generation of functional multipotent stem-like cells from terminally differentiated somatic cells without the introduction of retroviral mediated transgenes or ESC fusion.


Differences in pharmacological actions between beta-bungarotoxin and other neurotoxic phospholipases A2 purified from snake venoms.
Five highly toxic phospholipases A2 (PLAs) (beta-bungarotoxin, caudoxin, Mojave toxin, notexin and a basic PLA from Naja nigricollis venom) were compared for their pharmacological actions. In the chick biventer cervicis nerve-muscle preparation, all PLA toxins except beta-bungarotoxin (beta-BuTX) inhibited the postsynaptic acetylcholine response and induced contracture of the muscle at a high concentration. Indirect hemolytic activity was found in all PLA toxins and some of the toxins (Naja nigricollis basic PLA and Mojave toxin) even showed a potent direct hemolytic action, while beta-BuTX was devoid of both direct and indirect hemolytic activities on the guinea-pig erythrocytes. All PLA toxins except beta-BuTX caused an increase in muscle tone in the guinea-pig ileum at a concentration as low as 0.05 microgram/ml, and an increase in the contractile force in the guinea-pig atrium at a concentration of 1.0 microgram/ml. In contrast, beta-BuTX had no stimulant effect at concentrations up to 10 micrograms/ml. On the cultured cells, beta-BuTX suppressed the proliferation of neuroblastoma cells, but did not cause lysis of non-neuronal cells of the rat brain. However, beta-BuTX uniquely maintained a high population of viable cells in the neuroblastoma cell cultures. From these results it was concluded that beta-BuTX is the most specific presynaptic neurotoxin among the PLA toxins so far tested.


Association Between Homocysteine, Frailty and Biomechanical Response of the CNS in NPH-Suspected Patients.
Frailty is a geriatric syndrome that combines physiological decline, disruptions of homeostatic mechanisms across multiple physiologic systems and thus, strong vulnerability to further pathological stress. Previously, we provided the first evidence that increased risk of poor health outcomes, as quantified by a frailty index (FI), is associated with an alteration of the central nervous system (CNS) biomechanical response to blood pulsatility. In this study, we explored correlation between 14 biological parameters, the CNS elastance coefficient and FI. We included 60 adults (52-92 years) suspected of normal pressure hydrocephalus and presenting with markers of multiple coexisting brain pathologies, including Parkinson disease, Alzheimer disease, and vascular dementia. We showed that the homocysteine (Hcy) level was independently and positively associated with both the FI and the CNS elastance coefficient (adjusted R² of 10% and 6%). We also demonstrated that creatinine clearance and folate level were independently associated with Hcy level. Based on previous literature results describing the involvement of Hcy in endothelial dysfunction, glial activation, and neurodegeneration, we discuss how Hcy could contribute to the altered biomechanical response of the CNS and frailty.


Phase II open-label study of oral piritrexim in patients with advanced carcinoma of the urothelium who have experienced failure with standard chemotherapy.
Piritrexim is reported to have a response rate of 38% in patients with chemotherapy-naive disease and 23% for second-line therapy after chemotherapy failure. We report the results of a multiinstitutional, open-label, 2-stage, phase II study that further evaluates oral piritrexim in patients with urothelial carcinoma and who proved nonresponsive to standard chemotherapy. Eligible patients included those with bi-dimensionally measurable disease and an Eastern Cooperative Oncology Group performance status of 0-2, transitional cell carcinoma or adenocarcinoma of the urothelium, and nonresponse to > or = 1 previous standard chemotherapy regimen. Patients received piritrexim orally at 25 mg 3 times daily (every 8 hours regularly) for 5 consecutive days each week for 3 weeks, followed by a 1-week rest period. Treatment was continued until disease progression, unacceptable toxicity, or patient refusal. Of the 23 patients enrolled, 19 patients and 22 patients were assessable for toxicity and response, respectively. Two patients required dose reduction because of toxicity, 2 patients discontinued study because of toxicity, and 6 patients had > or = 1 serious adverse event. Except for grade 1/2 pain and fatigue, gastrointestinal toxicities were the most commonly reported events, followed by fever, delirium, and myelosuppression. No objective responses were observed, with 2 patients demonstrating stable disease after 2-4 cycles. By the statistical design of the trial, further enrollment was halted because of lack of activity. Regardless of modest side effects, oral piritrexim in heavily pretreated patients is inactive at this dose and schedule, confirming the results of a recent cooperative group trial.


COVID-19 and perceiving finitude: Associations with future time perspective, death anxiety, and ideal life expectancy.
The coronavirus pandemic threatens the health, future, and life of individuals and might hence accentuate perceptions of the fragility and finitude of life. We investigated how different perceptions of the pandemic (regarding the virus as a health threat and perceiving social and financial restrictions due to the pandemic) relate to different perceptions of life's finitude (i.e., future time perspective, death anxiety, and ideal life expectancy). Using longitudinal data from 1,042 adults (68% women; aged 18-95 years) gathered within the first and within the second peak of the pandemic in Germany, we expected decreases in future time perspective and ideal life expectancy, as well as increases in death anxiety in response to threatening perceptions of the pandemic. The results indicated decreasing future time perspectives, an accentuation of death anxiety right at the beginning of the pandemic, as well as stable ideal life expectancies. There was a tendency for more pronounced change among older adults. Initial levels and changes in the perceptions of finitude could partly be explained by initial and changing perceptions of the pandemic. Next to perceptions targeting the threat of the virus itself, perceptions of strong social and financial restrictions during the pandemic contributed to an altered stance toward the finitude of life. Concluding, we discuss stability and variation in perceptions of the finitude of life during a time of major societal change and a potentially life-threatening pandemic. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


Autologous and allogeneic ovarian orthotopic transplantation: morphologic, endocrinologic and natural pregnancy assessment.
To assess pregnancy of rabbits submitted to bilateral ovariectomy and orthotopic allogeneic or autologous intact and sliced ovarian transplantation without a vascular pedicle and to determine the morphofunctional aspects of the transplanted ovaries. Fifty-six female rabbits were studied. The ovaries were removed and orthotopically transplanted or replaced without vascular anastomoses: Group 1 (n=8), only laparotomy and laparorrhaphy, Group 2A (n=8) intact ovaries were transplanted on both sides, Group 2B (n=8) both ovaries were sliced and orthotopically transplanted, Group 2C (n=8), an intact ovary was transplanted on one side and a sliced ovary on the other side. In Group 3 the ovaries were reimplanted according to the procedure and subgroups described for Group 2. Three months later, the animals were paired with males for copulation. Estradiol, progesterone, FSH and LH hormone levels were assessed. A histologic study was carried out, and the number of pregnancies and litters were also determined. Chi-square test compared the number of pregnancies and litters. One-way ANOVA and the Tukey-Kramer tests compared the hormonal dosages. Pregnancies occurred in seven (87.5%) rabbits of Group 1, in three rabbits (37.5%) of Groups 2A and 3A, in four rabbits (50%) of groups 2B, 3B and 3C, and in five (62.5%) of group 2C. Normal hormone serum levels and histology confirmed the vitality of all ovaries. Orthotopic allogeneic and autologous ovarian transplantation without a vascular pedicle is viable in rabbits, and preserves their hormonal levels and fertile functions.


Genetic manipulation of adult-born hippocampal neurons rescues memory in a mouse model of Alzheimer's disease.
In adult mammals, neural progenitors located in the dentate gyrus retain their ability to generate neurons and glia throughout lifetime. In rodents, increased production of new granule neurons is associated with improved memory capacities, while decreased hippocampal neurogenesis results in impaired memory performance in several memory tasks. In mouse models of Alzheimer's disease, neurogenesis is impaired and the granule neurons that are generated fail to integrate existing networks. Thus, enhancing neurogenesis should improve functional plasticity in the hippocampus and restore cognitive deficits in these mice. Here, we performed a screen of transcription factors that could potentially enhance adult hippocampal neurogenesis. We identified Neurod1 as a robust neuronal determinant with the capability to direct hippocampal progenitors towards an exclusive granule neuron fate. Importantly, Neurod1 also accelerated neuronal maturation and functional integration of new neurons during the period of their maturation when they contribute to memory processes. When tested in an APPxPS1 mouse model of Alzheimer's disease, directed expression of Neurod1 in cycling hippocampal progenitors conspicuously reduced dendritic spine density deficits on new hippocampal neurons, to the same level as that observed in healthy age-matched control animals. Remarkably, this population of highly connected new neurons was sufficient to restore spatial memory in these diseased mice. Collectively our findings demonstrate that endogenous neural stem cells of the diseased brain can be manipulated to become new neurons that could allow cognitive improvement.


The first investigation of the synthetic mechanism and lithium intercalation chemistry of Li9Fe3(P2O7)3(PO4)2/C as cathode material for lithium ion batteries.
An iron-based mixed-polyanion compound, Li9Fe3(P2O7)3(PO4)2, is introduced as a possible cathode material for Li-ion batteries. Phase-pure Li9Fe3(P2O7)3(PO4)2 is successfully prepared by a sol-gel method, and its physicochemical properties are investigated in detail. Special attention is paid on making clear the variation of the phase composition with the annealing temperature and the effect of carbon coating on the electrochemical performance. Apparently phase-pure Li9Fe3(P2O7)3(PO4)2 can only be obtained in a narrow temperature range, either higher or lower annealing temperature outside this temperature range always leads to the impurity phase. The pristine Li9Fe3(P2O7)3(PO4)2 is suffering from its low electronic conductivity (10(-9) S cm(-1)) and theoretical capacity (85 mA h g(-1)), it has a first discharge capacity of only 36 mA h g(-1). Carbon coating is employed to improve the electrochemical performance. When the carbon content is 10 wt%, the discharge capacity of Li9Fe3(P2O7)3(PO4)2/C reaches the maximum value of 60 mA h g(-1). The electronic conductivity of the composite, the exact discharge capacity of Li9Fe3(P2O7)3(PO4)2 in the composite and the capacity retention of the composite after 30 cycles vary in the same fashion with an increase in carbon content, i.e. first quickly increase and then stabilize.


Redirecting AAV vectors to extrahepatic tissues.
Recombinant adeno-associated viral (AAV) vectors are the current benchmark for systemic delivery of gene therapies to multiple organs in vivo. Despite clinical successes, safe and effective gene delivery to extrahepatic tissues has proven challenging due to dose limiting toxicity arising from high liver uptake of AAV vectors. Deeper understanding of AAV structure, receptor biology, and pharmacology has enabled the design and engineering of liver-de-targeted capsids ushering in several new vector candidates. This next generation of AAVs offers significant promise for extrahepatic gene delivery to cardiovascular, musculoskeletal, and neurological tissues with improved safety profiles.


Postural stability after patient-specific interpositional knee spacer or total knee arthroplasty: a comparative study.
Total knee arthroplasty provides excellent results concerning functional demands, patient satisfaction and range of motion. Short-term results after implantation of patient-specific knee spacers also show encouraging results. We hypothesised that patient-specific interpositional devices provide better proprioception and postural stability by preserving the whole bone stock and both cruciate ligaments. In this study we compared functional results, proprioception and postural stability 16 months after 20 consecutive patient-specific interpositional device implantations of the knee (group A) and 20 consecutive total knee arthroplasties (group B). Patients were evaluated using the Knee Society Score (KSS) and the Western Ontario and McMaster Universities Arthritis Index (WOMAC). Postural stability was analysed during single leg stance on a force platform (Biodex). There were no significant differences between both groups concerning functional results 16 months after the procedure. Concerning postural stability, significant differences were found between both groups. Comparing the operated-on and non-operated-on sides in single leg stance, the operated-on leg gained the same stability as the non-operated-on side in both groups. The successful treatment of knee osteoarthritis can restore postural stability to the level of the unaffected side-independent from the implanted devices and with preservation of soft tissue and bone stock. Superior postural stability in joint-preserving patient-specific interpositional knee devices compared with total knee arthroplasty had to be attributed to the different age of both groups.


Serum magnesium and irradiation.
Serum magnesium determinations were obtained on 10 dogs and 11 patients undergoing fractionated irradiation to the pelvis and lower abdomen. Five of dogs received oral prednisone during irradiation. There was no significant change in magnesium concentration in either the control dogs or the patients, but there was a significant increase in stool frequency in both the dogs and patients. A significant increase in magnesium concentration was noted in the dogs receiving prednisone. It is concluded that radiation-induced diarrhea is not caused by reduced serum magnesium concentration.


AMP-activated protein kinase protects against anti-epidermal growth factor receptor-Pseudomonas exotoxin A immunotoxin-induced MA11 breast cancer cell death.
We have shown previously that our 425.3PE immunotoxin inhibits protein synthesis and induces apoptosis in human breast cancer cells. In attempts to further elucidate the intracellular pathways implicated in its cellular effects, we found that the immunotoxin induced an initial stress response, which rapidly caused an imbalance in the cellular energy status with an increase in reactive oxygen species. The AMP-activated protein kinase (AMPK), a sensor of increased cellular AMP/ATP ratio, was activated by 425.3PE. An immunotoxin-induced activation of c-Jun NH2-terminal kinase (JNK) preceded and overlapped caspase-mediated cleavage of the alpha-subunit of AMPK in a time- and dose-dependent manner. The JNK activation occurred already at a dose level too low to induce any detectable changes in the apoptotic machinery or protein synthesis. In contrast, cycloheximide, even at a concentration causing a 90% inhibition of protein synthesis, did neither affect the ATP level nor activate JNK and AMPK. Pretreatment of the cells with the specific AMPK inhibitor compound C and JNK inhibitor SP600125 blocked activation of AMPK and JNK, respectively, and subsequently sensitized the cells to 425.3PE-induced cell death. Whereas the antioxidant N-acetyl-l-cysteine blocked the generation of reactive oxygen species and activation of JNK and AMPK, it did not block immunotoxin-induced apoptosis. Together, the results show that 425.3PE induces several parallel signaling events, observed initially as an early activation of survival pathways, protecting the cells against the toxic effects of the immunotoxin, followed by subsequent apoptosis induction and protein synthesis inhibition. Conceivably, therapeutic manipulation of the signaling intermediates AMPK and JNK might provide a means to maximize the anticancer effects of the 425.3 immunotoxin.