Noninvasive fetal genotyping of paternally inherited alleles using targeted massively parallel sequencing in parentage testing cases.
Researchers have sought to develop a noninvasive protocol for paternity analysis that uses fetal cell-free DNA (cfDNA) in maternal plasma. Massively parallel sequencing (MPS) is expected to overcome this challenge because it enables the analysis of millions of DNA molecules at a single-base resolution. Seven women were involved in prenatal paternity testing cases. Before conventional invasive procedures, cfDNA was isolated from maternal plasma. Fetal tissues were then collected, as were blood samples from the alleged fathers. A custom array was designed that targeted 1497 regions containing single-nucleotide polymorphisms. These regions were massively parallel sequenced. In these seven cases, the mean nonmaternal allele fractions in maternal plasma ranged from 3.22% to 6.17%. Setting the allele fraction cutoff of 2.5%, 300 to 491 loci were considered informative for paternal origin and no genetic incompatibilities with the alleged fathers were found. These results were concordant with those of conventional short tandem repeat genotyping. Validation results performed using fetal samples showed that sequencing noise was completely filtered out, and 78.35% to 99.19% of the paternal alleles were accurately genotyped. The fetal cfDNA concentrations ranged from 7.12% to 13.81%, and the overall sequencing error rates ranged from 0.40% to 0.93%. In our study, we evaluate a straightforward method that can be used to identify paternal alleles based on analyses of paternal alleles and sequencing errors in maternal plasma. Our results support the notion that an MPS-based method could be utilized in noninvasive fetal genotyping and prenatal paternity analyses.


Molecular autopsy by proxy in preconception counseling.
Monogenic diseases that result in early pregnancy loss or neonatal death are genetically and phenotypically highly variable. This often poses significant challenges in arriving at a molecular diagnosis for reproductive planning. Molecular autopsy by proxy (MABP) refers to the genetic testing of relatives of deceased individuals to deduce the cause of death. Here, we specifically tested couples who lost one or more children/pregnancies with no available DNA. We developed our testing strategy using whole exome sequencing data from 83 consanguineous Saudi couples. We detected the shared carrier state of 50 pathogenic variants/likely pathogenic variants in 43 families and of 28 variants of uncertain significance in 24 families. Negative results were seen in 16 couples after variant reclassification. In 10 families, the risk of more than one genetic disease was documented. Secondary findings were seen in 10 families: either genetic variants with potential clinical consequences for the tested individual or a female carrier for X-linked conditions. This couple-based approach has enabled molecularly informed genetic counseling for 52% (43/83 families). Given the predominance of autosomal recessive causes of pregnancy and child death in consanguineous populations, MABP can be a helpful approach to consanguineous couples who seek counseling but lack molecular data on their deceased offspring.


Regioselective Bromination of Thieno[2',3':4,5]pyrrolo[1,2-d][1,2,4]triazin-8(7H)-one and Sequential Suzuki Couplings.
Regioselective bromination of thieno[2',3':4,5]pyrrolo[1,2-d][1,2,4]triazin-8(7H)-one at the 2- or 9-position was achieved by modulating the basicity of the reaction conditions. An anion-directed site-specific bromination mechanism was proposed. In addition, a one-pot bromination-Suzuki coupling protocol was developed for quick access of analogues at the 9-position.


Increase in cytosolic Ca2+ levels through the activation of non-selective cation channels induced by oxidative stress causes mitochondrial depolarization leading to apoptosis-like death in Leishmania donovani promastigotes.
Reactive oxygen species are important regulators of protozoal infection. Promastigotes of Leishmania donovani, the causative agent of Kala-azar, undergo an apoptosis-like death upon exposure to H2O2. The present study shows that upon activation of death response by H2O2, a dose- and time-dependent loss of mitochondrial membrane potential occurs. This loss is accompanied by a depletion of cellular glutathione, but cardiolipin content or thiol oxidation status remains unchanged. ATP levels are reduced within the first 60 min of exposure as a result of mitochondrial membrane potential loss. A tight link exists between changes in cytosolic Ca2+ homeostasis and collapse of the mitochondrial membrane potential, but the dissipation of the potential is independent of elevation of cytosolic Na+ and mitochondrial Ca2+. Partial inhibition of cytosolic Ca2+ increase achieved by chelating extracellular or intracellular Ca2+ by the use of appropriate agents resulted in significant rescue of the fall of the mitochondrial membrane potential and apoptosis-like death. It is further demonstrated that the increase in cytosolic Ca2+ is an additive result of release of Ca2+ from intracellular stores as well as by influx of extracellular Ca2+ through flufenamic acid-sensitive non-selective cation channels; contribution of the latter was larger. Mitochondrial changes do not involve opening of the mitochondrial transition pore as cyclosporin A is unable to prevent mitochondrial membrane potential loss. An antioxidant like N-acetylcysteine is able to inhibit the fall of the mitochondrial membrane potential and prevent apoptosis-like death. Together, these findings show the importance of non-selective cation channels in regulating the response of L. donovani promastigotes to oxidative stress that triggers downstream signaling cascades leading to apoptosis-like death.


Ultrastructural, immunohistological, and clinical findings in the pancreas in insulin-dependent diabetes mellitus (IDDM) of long duration.
Electron microscopical and immunohistological findings in small biopsies obtained at surgery from two subjects with longstanding type-I-diabetes [insulin-dependent diabetes mellitus (IDDM)] are described and demonstrated in relation to clinical data. The main findings are the first electron microscopical demonstration of A cells scattered as single cells in the exocrine pancreatic tissue and the detection of intermediate cells of the acinar-A cell type. Intermediate cells have never been reported before in the pancreas of diabetes in man.


Radionuclide (131)I-labeled multifunctional dendrimers for targeted SPECT imaging and radiotherapy of tumors.
We report the synthesis, characterization, and utilization of radioactive (131)I-labeled multifunctional dendrimers for targeted single-photon emission computed tomography (SPECT) imaging and radiotherapy of tumors. In this study, amine-terminated poly(amidoamine) dendrimers of generation 5 (G5·NH2) were sequentially modified with 3-(4'-hydroxyphenyl)propionic acid-OSu (HPAO) and folic acid (FA) linked with polyethylene glycol (PEG), followed by acetylation modification of the dendrimer remaining surface amines and labeling of radioactive iodine-131 ((131)I). The generated multifunctional (131)I-G5·NHAc-HPAO-PEG-FA dendrimers were characterized via different methods. We show that prior to (131)I labeling, the G5·NHAc-HPAO-PEG-FA dendrimers conjugated with approximately 9.4 HPAO moieties per dendrimer are noncytotoxic at a concentration up to 20 μM and are able to target cancer cells overexpressing FA receptors (FAR), thanks to the modified FA ligands. In the presence of a phenol group, radioactive (131)I is able to be efficiently labeled onto the dendrimer platform with good stability and high radiochemical purity, and render the platform with an ability for targeted SPECT imaging and radiotherapy of an FAR-overexpressing xenografted tumor model in vivo. The designed strategy to use the facile dendrimer nanotechnology may be extended to develop various radioactive theranostic nanoplatforms for targeted SPECT imaging and radiotherapy of different types of cancer.


Factors Affecting Quality of Life at Different Intervals After Treatment of Localized Prostate Cancer: Unique Influence of Treatment Decision Making Satisfaction, Personality and Sexual Functioning.
Using patient reported outcomes measures we identified the most informative set of factors associated with quality of life in a large sample of men treated for localized prostate cancer. We examined relationships with quality of life using FACIT (Functional Assessment of Chronic Illness Therapy). We also hypothesized variables in a sample of men diagnosed with localized prostate cancer who represented different time points since treatment, including less than 12 months in 70, 1 to 3 years in 344, greater than 3 to 5 years in 291 and greater than 5 years in 97. Correlative measures included subscales of MAX-PC (Memorial Anxiety Scale for Prostate Cancer), short forms of PROMIS® and SOMS (Surgical Outcomes Measurement System), TDM-SATS (Treatment Decision-Making Satisfaction Scale) and subscales of the BFI (Big Five Inventory) of personality. Quality of life was significantly associated with hypothesized variables across different time cohorts. In regression models several factors accounted for most of the variability in quality of life scores depending on time since treatment, including 47%, 22%, 29% and 27% at less than 12 months, 1 to 3 years, greater than 3 to 5 years and greater than 5 years, respectively. Upon examining the unique contribution of these variables, treatment decision making satisfaction was the only variable to have a significant and unique contribution to quality of life across all 4 time cohorts (standardized coefficients 0.33, 0.27, 0.31 and 0.49, respectively, p <0.01). In the cohort with 1 to 3 years since treatment erectile function and neurotic personality style also had unique associations with quality of life (standardized coefficients 0.25 and -0.20, respectively). When considering the short-term and the longer term quality of life of a man after treatment for localized prostate cancer, our findings highlight the importance of treatment decision making satisfaction, erectile function and personality.


Paraplegia after intercostal neurolysis with phenol.
In patients with advanced stages of cancer, severe pain is commonly encountered and is very difficult to treat. It affects the quality of life of the patient and the families involved. Pain can be managed using analgesics and adjuvant therapy. However, studies have shown that at least 10%-15% of patients fail to control pain adequately and will experience severe pain. We discuss the case of a 66-year-old female with metastatic adenoid cystic carcinoma of the left submandibular gland and developed paraplegia following intercostal neurolysis with phenol. After a successful diagnostic T6 to T12 intercostal nerve block, the patient was scheduled for an intercostal neurolytic block. We injected 2 mL of 10% aqueous phenol at each level on the left from the T6 to T12 ribs. One hour after the procedure, the patient developed bilateral lower extremity weakness with difficulty moving. A physical examination showed the absence of sensation to pinpricks and vibration from T10 to S5 and an absence of anal sphincter tone and sensation. Magnetic resonance images of the thoracic and lumbar spine showed leptomeningeal metastatic disease and myelitis. We postulate that the paraplegia could be from phenol diffusing along either the spinal nerves or the paravertebral venous plexus into the subarachnoid space. This case report points to the risks involved with phenol neurolysis close to the spine, and we propose alternative methods to minimize neurological complications.


What do hospices do? A survey of hospices in the United Kingdom and Republic of Ireland.
To obtain baseline information about hospice clinical activity. Survey of hospices by postal questionnaire and telephone interview. 98 Hospices in the United Kingdom and Republic of Ireland of 111 that had a named matron or senior nurse, including 17 funded by NHS and 81 independent units. Hospice matrons or nursing sisters in charge, to whom the questionnaire was addressed. Median age of hospices was 7 years, and those built during the past seven years had a median of 12 inpatient beds. All NHS hospices had some input from a medical consultant whereas 12 (15%) of independent units did not. 72 Hospices had home care teams and 12 of 20 of these randomly contacted by telephone provided 24 hour cover. The median number of whole time equivalent nurses was four, but 6 (30%) of teams did not include a doctor. Wide variations were found in discharge rates (range 1-76%) and throughput (1.7-31.8 patients/bed/year). In units with a full time consultant or medical director throughput was greater and more patients had palliative surgery and became organ donors than in units without (45/48 v 38/50 and 45/48 v 25/50, respectively). With the exception of pulmonary function tests and insertion of nasogastric tubes and indwelling epidural catheters, tests and procedures were used by over 90% of hospices, although sometimes the patient had to be transferred elsewhere. Respondents from units with a full time consultant or medical director were more likely to choose a "technical" description of their unit, such as "a pain relief centre" than those without, who favoured non-technical descriptions. These differences are likely to increase with the appointment of more fully trained consultants in palliative medicine.


Recent Advances Toward the Use of Mesoporous Silica Nanoparticles for the Treatment of Bacterial Infections.
It is a fact that the use of antibiotics is inducing a growing resistance on bacteria. This situation is not only the consequence of a drugs' misuse, but a direct consequence of a widespread and continuous use. Current studies suggest that this effect could be reversed by using abandoned antibiotics to which bacteria have lost their resistance, but this is only a temporary solution that in near future would lead to new resistance problems. Fortunately, current nanotechnology offers a new life for old and new antibiotics, which could have significantly different pharmacokinetics when properly delivered; enabling new routes able to bypass acquired resistances. In this contribution, we will focus on the use of porous silica nanoparticles as functional carriers for the delivery of antibiotics and biocides in combination with additional features like membrane sensitizing and heavy metal-driven metabolic-disrupting therapies as two of the most interesting combination therapies.


A mechanistic investigation on the utilization of lactose as a protective agent for multi-unit pellet systems.
The effect of lactose particle size on the extent of pellet coat damage was investigated. The extent of pellet coat damage increased linearly with lactose median particle size. It was observed that coated pellets compressed with coarser lactose grades had larger and deeper surface indentations. The surfaces of the pellets compressed with coarser lactose grades were also found to be significantly rougher. Micronized lactose was capable of protecting pellet coats from damage brought about by the presence of coarser lactose particles. The findings suggested a protective effect that micronized lactose conferred to pellet coats was not only through dimensional delimitations but also by higher interparticulate friction and longer particle rearrangement phase. As a result, the pellet volume fraction in the system was reduced. The extent of pellet coat damage was found to escalate when the pellet volume fraction in such system increased beyond a critical value of 0.39.


Probing temperature and capsaicin-induced activation of TRPV1 channel via computationally guided point mutations in its pore and TRP domains.
In a recent computational study, we revealed some mechanistic aspects of TRPV1 (transient receptor potential channel 1) thermal activation and gating and proposed a set of probable functionally important residues - "hot spots" that have not been characterized experimentally yet. In this work, we analyzed TRPV1 point mutants G643A, I679A + A680G, and K688G/P combining molecular modeling, biochemistry, and electrophysiology. The substitution G643A reduced maximal conductivity that resulted in a normal response to moderate stimuli, but a relatively weak response to more intensive activation. I679A + A680G channel was severely toxic for oocytes most probably due to abnormally increased basal activity of the channel ("always open" gates). The replacement K688G presumably facilitated movements of TRP domain and disturbed its coupling to the pore, thus leading to spontaneous activation and enhanced desensitization of the channel. Finally, mutation K688P was suggested to impair TRP domain directed movement, and the mutated channel showed ~100-fold less sensitivity to the capsaicin, enhanced desensitization and weaker activation by the heat. Our results provide a better understanding of TRPV1 thermal and capsaicin-induced activation and gating. These observations provide a structural basis for understanding some aspects of TRPV1 channel functioning and depict potentially pathogenic mutations.


[Rational use of psychotropic medications and sedative load in older adults with and without dementia].
Sedative drugs use has been associated with more cognitive impairment and increased mortality. Sedative load refers to cumulative exposure to multiple drugs with sedative properties. Describe the use of psychotropic drugs and sedative load in older adults with and without dementia. We conducted a cross-sectional study from 2014-2015 (Sanatorio Trinidad Mitre), in hospitalized patients older than 65 years old. Drugs were classified according to the WHO ATC system. The sedative load of drugs was calculated using the Linjakumpu model. 152 PsD and 35 PcD patients were registered, mean age 80.8±8.42. Polypharmacy was present in 44.39% being higher in patients with dementia than without dementia (62.80% vs 40.13%, p=0.0147). In 40.64% at least one psychotropic/sedative medication was used, greater in PcD (60% vs 36.18%, p=0.0097). The CS was: 1.32±1.59; 2.14 in PcD and 1.13 in PsD (p<0.001). Atypical antipsychotics and benzodiazepines were the most common (51.43 and 40% respectively) in patients without dementias. we evidenced a high level of prescription psychotropic or sedative drugs, mostly in patients with dementia. In those, the sedative load was greater. This finding highlights the importance of implementing strategies to optimize sedative drug use among older people.


Early epidermal changes in heat- and electrically injured pig skin. I. A light microscopic study.
In order to find methods applicable for disclosing electrical torture, pig skin was exposed to heat and electricity under controlled circumstances. Biopsies were obtained immediately after exposure and the morphology of the heat lesions was compared to that of electrical lesions. The cytoplasm of epidermal cells in heat lesions appeared granular or fibrillar, while the nuclei were rather unaffected. In electrical lesions the cytoplasm appeared homogeneous often with a peculiar white colour in hematoxylin-eosin stained sections. The nuclei were either dark and shadowy or enlarged and vesicular. The keratin in electrical lesions often showed a bright yellow colour. Small defects in the epidermis were seen in some electrical lesions. Thus the morphology of electrical lesions differed markedly from that of heat lesions. Although some of these differences may be due to differences in distribution and intensity of energy, it is probable that pH shifts in the cells due to electrolysis is the main cause of the specific morphology of electrical lesions.


Intermediate Type of Juvenile Paget's Disease: A Rare Case in Indian Population.
Juvenile Paget's disease (JPD), a rare genetic skeletal disorder characterized by accelerated bone turnover with elevated levels of serum alkaline phosphatase, presents in early childhood. We report a female patient with typical features of JPD with dental finding who remained undiagnosed until 18 years of age. Scarcity of this disease in the Indian literature and need for timely diagnosis to avert progression of disease thus incited us to report this case.


Septris: a novel, mobile, online, simulation game that improves sepsis recognition and management.
Annually affecting over 18 million people worldwide, sepsis is common, deadly, and costly. Despite significant effort by the Surviving Sepsis Campaign and other initiatives, sepsis remains underrecognized and undertreated. Research indicates that educating providers may improve sepsis diagnosis and treatment; thus, the Stanford School of Medicine has developed a mobile-accessible, case-based, online game entitled Septris (http://med.stanford.edu/septris/). Septris, launched online worldwide in December 2011, takes an innovative approach to teaching early sepsis identification and evidence-based management. The free gaming platform leverages the massive expansion over the past decade of smartphones and the popularity of noneducational gaming.The authors sought to assess the game's dissemination and its impact on learners' sepsis-related knowledge, skills, and attitudes. In 2012, the authors trained Stanford pregraduate (clerkship) and postgraduate (resident) medical learners (n = 156) in sepsis diagnosis and evidence-based practices via 20 minutes of self-directed game play with Septris. The authors administered pre- and posttests. By October 2014, Septris garnered over 61,000 visits worldwide. After playing Septris, both pre- and postgraduate groups improved their knowledge on written testing in recognizing and managing sepsis (P < .001). Retrospective self-reporting on their ability to identify and manage sepsis also improved (P < .001). Over 85% of learners reported that they would or would maybe recommend Septris. Future evaluation of Septris should assess its effectiveness among different providers, resource settings, and cultures; generate information about how different learners make clinical decisions; and evaluate the correlation of game scores with sepsis knowledge.


CEST imaging of fast exchanging amine pools with corrections for competing effects at 9.4 T.
Chemical exchange saturation transfer (CEST) imaging of fast exchanging amine protons at 3 ppm offset from the water resonant frequency is of practical interest, but quantification of fast exchanging pools by CEST is challenging. To effectively saturate fast exchanging protons, high irradiation powers need to be applied, but these may cause significant direct water saturation as well as non-specific semi-solid magnetization transfer (MT) effects, and thus decrease the specificity of the measured signal. In addition, the CEST signal may depend on the water longitudinal relaxation time (T1w ), which likely varies between tissues and with pathology, further reducing specificity. Previously, an analysis of the asymmetry of saturation effects (MTRasym ) has been commonly used to quantify fast exchanging amine CEST signals. However, our results show that MTRasym is greatly affected by the above factors, as well as asymmetric MT and nuclear Overhauser enhancement (NOE) effects. Here, we instead applied a relatively more specific inverse analysis method, named AREX (apparent exchange-dependent relaxation), that has previously been applied only to slow and intermediate exchanging solutes. Numerical simulations and controlled phantom experiments show that, although MTRasym depends on T1w and semi-solid content, AREX acquired in steady state does not, which suggests that AREX is more specific than MTRasym . By combining with a fitting approach instead of using the asymmetric analysis to obtain reference signals, AREX can also avoid contaminations from asymmetric MT and NOE effects. Animal experiments show that these two quantification methods produce differing contrasts between tumors and contralateral normal tissues in rat brain tumor models, suggesting that conventional MTRasym applied in vivo may be influenced by variations in T1w , semi-solid content, or NOE effect. Thus, the use of MTRasym may lead to misinterpretation, while AREX with corrections for competing effects likely enhances the specificity and accuracy of quantification to fast exchanging pools.


3D myocardial T1 mapping at 3T using variable flip angle method: pilot study.
Myocardial T1 mapping is an emerging technique that could improve cardiovascular magnetic resonance diagnostic accuracy. In this study, a variable flip angle approach with B1 correction is proposed at 3T on the myocardium, employing standard 3D spoiled fast gradient echo and echo planar imaging sequences. The method was tested on phantoms to determine the set of standard 3D spoiled fast gradient echo angles adapted to myocardial T1 measurements and was compared to the inversion-recovery spin-echo reference T1 method. Seven volunteers underwent magnetic imaging resonance to acquire myocardial T1 maps and T1 values of the human heart. This original method demonstrated good reproducibility in phantoms and a significant correlation between variable flip angle T1 values and reference inversion-recovery spin-echo T1 values. It yielded myocardial T1 values consistent with expected T1 and an increasing homogenization of myocardial segments owing to B1 correction. The mean myocardial T1 value was 1341 ± 42 ms. Myocardial 3D T1 mapping using the variable flip angle approach can potentially be useful for evaluating fibrosis on the entire myocardium using a standard clinical sequence.


Preschool competency in context: an investigation of the unique contribution of child competencies to early academic success.
The present concurrent study combined developmental and ecological considerations to examine the unique contribution of multiple preschool competencies to an indicator of early academic success. Participants included 195 Head Start children from 32 classrooms representative of a large, urban Head Start program. Dimensional (variable-centered) analyses revealed 3 distinct classroom competency dimensions (i.e., General Classroom Competencies, Specific Approaches to Learning, and Interpersonal Classroom Behavioral Problems). The first 2 of these dimensions were found to be uniquely associated with early academic success. Findings from typological (person-centered) analyses supported the dimensional findings. Typological analyses revealed 7 profiles of classroom competency distinguished by high scores on the dimensions of General Competencies and Approaches to Learning, and these profiles were found to relate differentially to the indicator of early academic success.


Unilateral Posterior Polymorphous Corneal Dystrophy Presented as Anisometropic Astigmatism: 3 Case Reports.
Posterior polymorphous corneal dystrophy (PPCD) is typically considered bilateral and asymptomatic. However, few case reports on patients with unilateral PPCD with asymmetric refractive error have mentioned anisometropic amblyopia development. In support of this, we report 3 cases of unilateral PPCD that presented as anisometropic astigmatism. Visual prognosis related to amblyopia development is discussed. All 3 patients had a band lesion in the affected eye and a difference of at least 1.5 diopters in cylindrical refractive error between their eyes. The affected eye had a greater amount of astigmatism in all cases. Two patients (Cases 1 and 2) also had amblyopia in the affected eye. Case 1 was a 25-year-old male with a unilateral PPCD diagnosis and a band lesion involving the visual axis. Case 2 was an 11-year-old boy diagnosed with unilateral PPCD. The boy was treated with occlusion and atropine therapy over a 2-year period. Case 3 was a 4-year-old girl diagnosed with unilateral PPCD. The girl had a 30-month history of corrective spectacle use and had no amblyopia. In all cases, the corneal endothelial cell count was lower in the affected eye than in the unaffected contralateral eye. Practitioners should closely monitor patients with unilateral PPCD for astigmatic anisometropia and amblyopia development. Visual prognosis for patients with unilateral PPCD may be related to lesion position, age at diagnosis, astigmatism severity, and early-childhood corrective spectacle use.


Changing models of biomedical research.
Thanks to the revolutions in information technology, human "-omics" research, and intricate medical device development, academic health centers (AHCs) now have an unparalleled potential to become translational engines that both generate basic science advances and then translate them into human studies and, ultimately, into improved health care. However, AHC infrastructures have traditionally been optimized to foster basic research. Now modifications must be made to meet these expanded roles of AHCs, from providing fundamental biomedical insights to first-in-human interventions and, if warranted, to larger randomized clinical trials. Eventually, AHCs must integrate these improved treatments into patient care. Challenges to this process have been defined by the Institute of Medicine. Building the appropriate infrastructures for human investigation and stabilizing the careers of young physicians essential to these translational events have become critical needs that will require systemic investments if AHCs are to respond to these biological revolutions and fulfill their promise.


Presence of neutralizing antibodies to Orthopoxvirus in capybaras (Hydrochoerus hydrochaeris) in Brazil.
Cantagalo virus is a strain of vaccinia virus (genus Orthopoxvirus) and the etiological agent of an important vesicopustular disease that affects dairy cows and milkers in Brazil. The reservoirs involved in the maintenance of this virus in nature are unknown. In the present work, the detection of neutralizing antibodies to Orthopoxvirus in capybaras collected in São Paulo state is reported. Capybaras are the largest rodent species native to South America and have already been reported as putative reservoirs of other pathogenic microorganisms. Thirteen out of thirty-three serum samples were found positive in plaque-reduction neutralization tests, some of them showing high titers compared to positive controls. These results suggest that capybaras may play a role in the infection cycle of vaccinia virus in Brazil.


Emerging avian influenza infections: Current understanding of innate immune response and molecular pathogenesis.
The highly pathogenic avian influenza viruses (HPAIVs) cause severe disease in gallinaceous poultry species, domestic ducks, various aquatic and terrestrial wild bird species as well as humans. The outcome of the disease is determined by complex interactions of multiple components of the host, the virus, and the environment. While the host-innate immune response plays an important role for clearance of infection, excessive inflammatory immune response (cytokine storm) may contribute to morbidity and mortality of the host. Therefore, innate immunity response in avian influenza infection has two distinct roles. However, the viral pathogenic mechanism varies widely in different avian species, which are not completely understood. In this review, we summarized the current understanding and gaps in host-pathogen interaction of avian influenza infection in birds. In first part of this article, we summarized influenza viral pathogenesis of gallinaceous and non-gallinaceous avian species. Then we discussed innate immune response against influenza infection, cytokine storm, differential host immune responses against different pathotypes, and response in different avian species. Finally, we reviewed the systems biology approach to study host-pathogen interaction in avian species for better characterization of molecular pathogenesis of the disease. Wild aquatic birds act as natural reservoir of AIVs. Better understanding of host-pathogen interaction in natural reservoir is fundamental to understand the properties of AIV infection and development of improved vaccine and therapeutic strategies against influenza.


Examination of the best pressure range for ion/molecule reactions of anthraquinones in an external source ion trap mass spectrometer.
This study outlines some observations of the pressure effect for gas phase ion-molecule reactions of anthraquinone derivatives with dimethyl ether in an external source ion trap mass spectrometer. At the reagent pressure of 7.998 x 10(-2) Pa, formation of the protonated ions, [M + 13]+, [M + 15]+, and [M + 45]+ ions, of anthraquinones can be observed. However, at the pressure of 1.066 x 10(-2) Pa, formation of molecular ions and many fragment ions of the M+. or [M + H]+ ions have been observed. Since the pressure effect is notable within a small range of pressures for many compounds, it is important to draw attention to the use of the ion trap with an external source where other factors such as ion source residence time may play a role. This can also provide some information for better and more careful controls of the reagent pressure in order to obtain fair CI spectra in an external source ion trap mass spectrometer.


[The Pacemaker and Implantable Cardioverter-Defibrillator Registry of the Italian Association of Arrhythmology and Cardiac Pacing - Report 2020].
The Pacemaker (PM) and Implantable Cardioverter-Defibrillator (ICD) Registry of the Italian Association of Arrhythmology and Cardiac Pacing (AIAC) collects information about demographics, clinical characteristics, main indications for PM/ICD therapy and device types from the Italian collaborating centers. For the year 2020, the main data about national PM and ICD implantation activity were obtained on the basis of European Cards and subsequently analyzed to create a standard report. PM Registry: data about 22 080 PM implantations were collected (18 027 first implants and 3803 replacements). The number of collaborating centers was 142. Median age of treated patients was 82 years (75 quartile I; 87 quartile III). ECG indications included atrioventricular conduction disorders in 31.0% of first PM implants, sick sinus syndrome in 13.8%, atrial fibrillation plus bradycardia in 9.9%, other unspecified ECG and electrophysiological abnormalities in 36.6%. Use of single-chamber PMs was reported in 29.0% of first implants, of dual-chamber PMs in 68.2%, of PM with cardiac resynchronization therapy (CRT) in 2.7%. ICD Registry: data about 11 931 ICD implantations were obtained (8266 first implants and 3665 replacements). The number of collaborating centers was 330. Median age of treated patients was 72 years [63 quartile I; 79 quartile III]. Primary prevention indication was reported in 85.0% of first implants, secondary prevention in 15.0% (cardiac arrest in 4.2%). A single-chamber ICD was used in 30.2% of first implants, dual-chamber ICD in 31.2% and biventricular ICD in 38.6%. In the calendar year 2020, the Italian PM Registry showed stable ECG and symptom indications, with an important prevalence of dual-chamber pacing. The ICD Registry documented a large use of prophylactic and biventricular ICDs, reflecting a favorable adherence in clinical practice to trials and guidelines. The ICD longevity and the number of recalls demonstrated a favorable trend. In order to increase and optimize the cooperation of Italian PM and ICD implanting centers, the online data entry (https://www.aiac.it/riprid) should be adopted at large scale.


Recent trend in the choice of fillers and injection techniques in Asia: a questionnaire study based on expert opinion.
With recognition of the value of volume enhancement in achieving a more youthful appearance, there has been concomitant explosion in the soft tissue filler market. Given the vast array of filler products and techniques currently available, choosing the right product and technique can be overwhelming to those with little experience. To evaluate the recent trend in the choice of fillers and injection techniques among leading dermatologists in Asia and offer guidance to those who practice facial fillers. A panel of dermatologists, who are recognized as filler experts and key speakers in Korea were asked to fill out an in-depth questionnaire on fillers in 2012. The results of the 2012 questionnaire are presented and compared with the questionnaire results of the exact same group of doctors in 2011. Those who participated in the questionnaire study practiced fillers for an average of 10.6 years with an average of 32.8 filler cases per week. Common indications for filler injection were midface augmentation and nose augmentation. Indications that most drastically increased between 2011 and 2012 were midface and forehead augmentation. For the nasolabial folds, the most preferred choice of filler product, needle, injection technique and injection depth was Radiesse®, 27G short needle, Layering technique and the Upper subcutaneous fat layer. For filler rhinoplasty, the preferred choices were Radiesse®, 27G short needle, Linear threading technique and the Mid-deep fatty layer. For dark circles, the favored choices were Esthelis Basic®, 30G short needle, Vertical technique and the SOOF (suborbicularis oculi fat) layer. For forehead augmentation, the most favored choices were Juvederm Voluma®, 23G cannula, Linear threading technique and Fanning and the Supraperiosteal layer. The physicians' satisfaction score for the nasolabial folds, filler rhinoplasty, dark circles and forehead augmentation was 71.5, 90, 84.5 and 87 respectively. On general, filler experts preferred fillers with relatively high visco-elasticity for the nasolabial folds, nose augmentation and forehead augmentation but chose fillers with low visco-elasticity for dark circles. Linear treading technique (with or without fanning) was universally popular but Vertical injection was considered more useful for dark circles and the nasal tip.


Azithromycin and erythromycin susceptibility and macrolide resistance genes in Prevotella from patients with periodontal disease.
To study oral Prevotella spp. isolated from patients with chronic periodontitis, to determine their susceptibility to azithromycin and erythromycin and to screen the presence of macrolide resistance genes therein. Isolates with a Prevotella-like morphology were obtained from subgingival samples of 52 patients with chronic periodontitis. Each isolate was identified to the species level by sequencing of the 16S rRNA gene. In 100 Prevotella spp. isolates, azithromycin and erythromycin susceptibility was determined using the E test method, and the screening of erm(A), erm(B), erm(C), erm(F), erm(G), erm(Q) and mef(A) genes was done by PCR. Prevotella intermedia and Prevotella nigrescens were the most identified species (33% each). Minimum inhibitory concentrations (MICs) ranges for both antibiotics were 0.016/0.032 to >256 μg/ml. MIC50 values for azithromycin and erythromycin were 1.5 and 1 μg/ml, respectively, and MIC90 values were >256 μg/ml for both antibiotics. Nineteen per cent of the isolates carried erm(B), and 51% carried erm(F). The MIC values found were high compared to previous studies. erm(F) was greatly prevalent, and we describe for the first time the erm(B) gene in Prevotella spp. The presence of either of the genes seems to be associated with a higher degree of resistance to azithromycin and erythromycin.


Registered nurses' perception of self-efficacy and competence in smoking cessation after participating in a web-based learning activity.
To describe how registered nurses having undergone a web-based learning activity perceive their self-efficacy and competence to support patients with smoking cessation in connection with surgery. Smoking cessation in connection with surgery reduces postoperative complications, and the support patients get from registered nurses may be important in helping them become smoke-free in connection with their surgery. Therefore, registered nurses are in need of enhanced understanding about which kind of counselling is the most effective for smoking cessation. Educating large groups of registered nurses in a digital environment appears to be a flexible and cost-effective way. A convergent mixed-method design with data collection was done using questionnaires (n = 47) and semistructured interviews (n = 11). Inclusion criteria were registered nurses in surgical wards. The samples were nonprobability and modified nested. Descriptive statistics and content analysis were used for data analysis. After completing the web-based learning activity, the registered nurses perception was that of good self-efficacy and increased competence in supporting patients with smoking cessation in connection with surgery. They improved their understanding of how to talk about smoking cessation with patients in dialogue using open-ended questions. Nevertheless, the registered nurses requested opportunities for dialogue and interaction with colleagues or topic experts. The results indicate that registered nurses can enhance their competence in supporting patients to embrace smoking cessation by learning in a digital environment. Self-efficacy and understanding of the topic seems to motivate registered nurses to counsel patients about smoking cessation. Findings from this study will be of particular interest to educators in healthcare settings who can devise further development of web-based learning activities.


Recent advances in CAR-T cell engineering.
Chimeric antigen receptor T (CAR-T) cell therapy is regarded as an effective solution for relapsed or refractory tumors, particularly for hematological malignancies. Although the initially approved anti-CD19 CAR-T therapy has produced impressive outcomes, setbacks such as high relapse rates and resistance were experienced, driving the need to discover engineered CAR-T cells that are more effective for therapeutic use. Innovations in the structure and manufacturing of CAR-T cells have resulted in significant improvements in efficacy and persistence, particularly with the development of fourth-generation CAR-T cells. Paired with an immune modifier, the use of fourth-generation and next-generation CAR-T cells will not be limited because of cytotoxic effects and will be an efficient tool for overcoming the tumor microenvironment. In this review, we summarize the recent transformations in the ectodomain, transmembrane domain, and endodomain of the CAR structure, which, together with innovative manufacturing technology and improved cell sources, improve the prospects for the future development of CAR-T cell therapy.


A truncated IL-12rβ1 receptor ameliorates chronic graft-versus-host disease-induced lupus nephritis by inhibiting Th1 and Th17 cells.
Th1 and Th17 cells have been strongly implicated in the pathogenesis of systemic lupus erythematosus (SLE). Interleukin (IL)-12 and IL-23 respectively drive the polarization of Th1 and Th17 cells and share a common p40 subunit. In this study, the protective and therapeutic effects of a truncated human IL-12rβ1 receptor (tIL12rβ1) targeting IL-12/IL-23 p40 were evaluated in chronic graft-versus-host disease (cGVHD)-induced SLE-like model. The results indicated that tIL12rβ1 treatment effectively delayed the proteinuria onset and induced a significant remission of proteinuria, autoantibody production, and immune complex deposition in the mouse model. Remarkably, the therapeutic effects of tIL12rβ1 were predominantly dependent on the suppression of pathogenic Th1 and Th17 cell commitment through the reduction of RORγt and T-bet expression. Collectively, this receptor molecule may offer a new treatment option for SLE.


[A brief history of the application of urethral catheterization in ancient China].
Through analysing the ancient medical literatures, the author described the types and the history of urethral catheterization in ancient China and claims that, since urethral catheterization had been used in the Jin dynasty, Sun Simiao isn't the first doctor who used this technique.


A geometric principle may guide self-assembly of fullerene cages from clathrin triskelia and from carbon atoms.
Clathrin triskelia and carbon atoms alike self-assemble into a limited selection of fullerene cages (with n three connected vertices, 3n/2 edges, 12 pentagonal faces, and (n-20)/2 hexagonal faces). We show that a geometric constraint-exclusion of head-to-tail dihedral angle discrepancies (DADs)-explains this limited selection as well as successful assembly into such closed cages in the first place. An edge running from a pentagon to a hexagon has a DAD, since the dihedral angles about the edge broaden from its pentagon (tail) end to its hexagon (head) end. Of the 21 configurations of a central face and surrounding faces, six have such DAD vectors arranged head-to-tail. Of the 5770 mathematically possible fullerene cages for n <or= 60, excluding those with any of the six configurations leaves just 15 cages plus buckminsterfullerene (n = 60), among them the known clathrin cages. Of the 216,739 mathematically possible cages for 60 < n <or= 84, just the 50 that obey the isolated-pentagon rule, among them known carbon cages, pass. The absence of likely fullerenes for some n (30,34,46,48,52-58,62-68) explains the abundance of certain cages, including buckminsterfullerene. These principles also suggest a "probable roads" path to self-assembly in place of pentagon-road and fullerene-road hypotheses.


Magnetic resonance imaging of the shoulder in abduction.
To simulate the painful arm position in subacromial impingement syndrome, magnetic resonance imaging of the shoulder was done with the arms at the sides and at 80 degrees abduction in two groups of patients. Group A had 20 shoulders in 20 patients with subacromial impingement syndrome and Group B had 19 shoulders in 19 patients without impingement syndrome. When arm position was changed from adduction to abduction, an increase in signal intensity of the rotator cuff tendon was seen more often in Group A (45%) than in Group B (26%). Group A shoulders showed encroachment of the acromion or the acromioclavicular joint to the rotator cuff in 25% of the shoulders with the arms at the sides, and in 75% with the arms in abduction. Similar findings were observed in only 14% and 21% of Group B shoulders in the respective arm positions. Abduction of the arms seemed to cause encroachment of the overlying structure, especially of the acromioclavicular joint, to the rotator cuff in patients with subacromial impingement syndrome, even if there were no such findings in the images with the arms at the sides.


[Bacterial infections in patients with liver cirrhosis and ascites].
Bacterial infections are common complications and the cause of death in patients with cirrhosis and ascites. There is no standard method for a rapid and low-cost diagnosis, and its prognosis is poor. The aim of this study was to determine the etiology and frequency of bacterial infections in patients with liver cirrhosis of different etiology, and the influence of bacterial infections on the prognosis in patients with liver cirrhosis and ascites. Sixty-four patients with cirrhosis and ascites were included in the study. The diagnosis of spontaneous bacterial peritonitis was established based on the diagnostic abdominal paracentesis and the results of biochemical, cytological and microbiologic analysis of ascitic fluid. The diagnosis of urinary infection and pneumonia were made according to the standard criteria. Spontaneous bacterial peritonitis was diagnosed in 23 (35.9%) patients, urinary infections in 16(25%) and pneumonia in 11 (17.2%). Gram positive and gram negative bacteria in spontaneous bacterial peritonitis were etiologically almost equally represented (52%; 48%).The most frequent causes were Escherichia coli and Staphylococcus aureus. In 81% of patients urinary infections were caused by gram negative bacteria (Escherichia coli in 44%). The most frequent cause of pneumonia was Streptococcus pneumoniae (46%). Spontaneous bacterial peritonitis, urinary infections and bronchopneumonia are the most frequent bacterial infections in patients with liver cirrhosis and ascites. A timely recognition of bacterial infections and the initiation of treatment have a positive effect on the prognosis of such patients.


Pump-degenerate four wave mixing as a technique for analyzing structural and electronic evolution: multidimensional time-resolved dynamics near a conical intersection.
Pump-degenerate four wave mixing (pump-DFWM) is used to simultaneously study the early events in structural and electronic population dynamics of the non-adiabatic passage between two excited electronic states. After the precursor state S2 is populated by an initial pump beam, a DFWM sequence is set resonant with the S1 --> Sn transition on the successor state S1. The information obtained by pump-DFWM is two-fold: by scanning the delay between the initial pump and the DFWM sequence, the evolution of the individual excited-state modes is observed with a temporal resolution of 20 fs and a spectral resolution of 10 cm-1. Additionally, pump-DFWM yields information on electronic population dynamics, resulting in a comprehensive description of the S2 --> S1 internal conversion. As a system in which the interplay between structural and electronic evolution is of great interest, all-trans-beta-carotene in solution was chosen. The pump-DFWM signal is analyzed for different detection wavelengths, yielding results on the ultrafast dynamics between 1Bu+ (S2) and 2Ag- (S1). The process of vibrational cooling on S1 is discussed in detail. Furthermore, a low-lying vibrationally hot state is excited and characterized in its spectroscopic properties. The combination of highly resolved vibrational dynamics and simultaneously detected ultrafast electronic state spectroscopy gives a complete picture of the dynamics near a conical intersection. Because pump-DFWM is a pure time domain technique, it offers the prospect of coherent control of excited-state dynamics on an ultrafast time scale.


Fine-tuning the ubiquitin code at DNA double-strand breaks: deubiquitinating enzymes at work.
Ubiquitination is a reversible protein modification broadly implicated in cellular functions. Signaling processes mediated by ubiquitin (ub) are crucial for the cellular response to DNA double-strand breaks (DSBs), one of the most dangerous types of DNA lesions. In particular, the DSB response critically relies on active ubiquitination by the RNF8 and RNF168 ub ligases at the chromatin, which is essential for proper DSB signaling and repair. How this pathway is fine-tuned and what the functional consequences are of its deregulation for genome integrity and tissue homeostasis are subject of intense investigation. One important regulatory mechanism is by reversal of substrate ubiquitination through the activity of specific deubiquitinating enzymes (DUBs), as supported by the implication of a growing number of DUBs in DNA damage response processes. Here, we discuss the current knowledge of how ub-mediated signaling at DSBs is controlled by DUBs, with main focus on DUBs targeting histone H2A and on their recent implication in stem cell biology and cancer.


Nanopore sequencing technology and its applications.
Since the development of Sanger sequencing in 1977, sequencing technology has played a pivotal role in molecular biology research by enabling the interpretation of biological genetic codes. Today, nanopore sequencing is one of the leading third-generation sequencing technologies. With its long reads, portability, and low cost, nanopore sequencing is widely used in various scientific fields including epidemic prevention and control, disease diagnosis, and animal and plant breeding. Despite initial concerns about high error rates, continuous innovation in sequencing platforms and algorithm analysis technology has effectively addressed its accuracy. During the coronavirus disease (COVID-19) pandemic, nanopore sequencing played a critical role in detecting the severe acute respiratory syndrome coronavirus-2 virus genome and containing the pandemic. However, a lack of understanding of this technology may limit its popularization and application. Nanopore sequencing is poised to become the mainstream choice for preventing and controlling COVID-19 and future epidemics while creating value in other fields such as oncology and botany. This work introduces the contributions of nanopore sequencing during the COVID-19 pandemic to promote public understanding and its use in emerging outbreaks worldwide. We discuss its application in microbial detection, cancer genomes, and plant genomes and summarize strategies to improve its accuracy.


Single-molecule detection of airborne singlet oxygen.
Airborne singlet oxygen (1O2) molecules, which are generated during the TiO2 photocatalytic reactions and diffused from the surface into air, were detected at the opposite surface using terrylenediimide (TDI) molecules at the single-molecule level. The novel 1O2 nanosensor, which has a detectable number of about 1000 1O2 molecules in 70 x 70 square micrometers, can easily detect the single 1O2 molecule at a distance of over 1000 micrometers from the place of its creation in ambient air.


Compact FPGA hardware architecture for public key encryption in embedded devices.
Security is a crucial requirement in the envisioned applications of the Internet of Things (IoT), where most of the underlying computing platforms are embedded systems with reduced computing capabilities and energy constraints. In this paper we present the design and evaluation of a scalable low-area FPGA hardware architecture that serves as a building block to accelerate the costly operations of exponentiation and multiplication in [Formula: see text], commonly required in security protocols relying on public key encryption, such as in key agreement, authentication and digital signature. The proposed design can process operands of different size using the same datapath, which exhibits a significant reduction in area without loss of efficiency if compared to representative state of the art designs. For example, our design uses 96% less standard logic than a similar design optimized for performance, and 46% less resources than other design optimized for area. Even using fewer area resources, our design still performs better than its embedded software counterparts (190x and 697x).


Mechanism for ginkgolic acid (15 : 1)-induced MDCK cell necrosis: Mitochondria and lysosomes damages and cell cycle arrest.
Ginkgolic acids (GAs), primarily found in the leaves, nuts, and testa of ginkgo biloba, have been identified with suspected allergenic, genotoxic and cytotoxic properties. However, little information is available about GAs toxicity in kidneys and the underlying mechanism has not been thoroughly elucidated so far. Instead of GAs extract, the renal cytotoxicity of GA (15 : 1), which was isolated from the testa of Ginkgo biloba, was assessed in vitro by using MDCK cells. The action of GA (15 : 1) on cell viability was evaluated by the MTT and neutral red uptake assays. Compared with the control, the cytotoxicity of GA (15 : 1) on MDCK cells displayed a time- and dose-dependent manner, suggesting the cells mitochondria and lysosomes were damaged. It was confirmed that GA (15 : 1) resulted in the loss of cells mitochondrial trans-membrane potential (ΔΨm). In propidium iodide (PI) staining analysis, GA (15 : 1) induced cell cycle arrest at the G0/G1 and G2/M phases, influencing on the DNA synthesis and cell mitosis. Characteristics of necrotic cell death were observed in MDCK cells at the experimental conditions, as a result of DNA agarose gel electrophoresis and morphological observation of MDCK cells. In conclusion, these findings might provide useful information for a better understanding of the GA (15 : 1) induced renal toxicity.


A dangerous cough.
Patient X is a 48-year-old retired potteries worker who presented to his general practitioner (GP) with a non-productive cough, which had lasted over 1 month, and it was a television advert that he had seen that prompted him to visit his doctor. A cough is the most common presenting complaint for patients when they visit their GP and when considering differential diagnoses it is important to look at a patient's medical, drug and social history. It would have been easy for the GP to dismiss the cough as an exacerbation of the patient's asthma based on his medical history, but on further questioning into the patient's social history an emergency chest x-ray was ordered, which later revealed a bronchial carcinoma. This case highlights the importance of the media in healthcare advertising and describes how a fairly innocent presentation can later turn out to be a more sinister pathology.


Herbicidal properties of antimalarial drugs.
The evolutionary relationship between plants and the malarial parasite Plasmodium falciparum is well established and underscored by the P. falciparum apicoplast, an essential chloroplast-like organelle. As a result of this relationship, studies have demonstrated that herbicides active against plants are also active against P. falciparum and thus could act as antimalarial drug leads. Here we show the converse is also true; many antimalarial compounds developed for human use are highly herbicidal. We found that human antimalarial drugs (e.g. sulfadiazine, sulfadoxine, pyrimethamine, cycloguanil) were lethal to the model plant Arabidopsis thaliana at similar concentrations to market herbicides glufosinate and glyphosate. Furthermore, the physicochemical properties of these herbicidal antimalarial compounds were similar to commercially used herbicides. The implications of this finding that many antimalarial compounds are herbicidal proffers two novel applications: (i) using the genetically tractable A. thaliana to reveal mode-of-action for understudied antimalarial drugs, and (ii) co-opting antimalarial compounds as a new source for much needed herbicide lead molecules.


Current internal exposure to pesticides in children and adolescents in Germany: urinary levels of metabolites of pyrethroid and organophosphorus insecticides.
Pesticides are widely used throughout the world, in agriculture to protect crops and in public health to control diseases transmitted by vectors or intermediate hosts. After the prohibition of organochlorines such as DDT, today, mainly pyrethroids and organophosphorus insecticides are used. Whereas many studies have been published on background exposure of the population to organochlorines, data on internal exposure of the population to pyrethroids and organophosphorus insecticides are scarce. Here, we report on internal exposure of children and young people, in an urban area in Germany, to pyrethroids and organophosphorus acids, assessed by the analysis of urinary levels of their corresponding specific metabolites. Approximately 673 children and adolescents took part in this voluntary investigation, including 331 children <6 years of age. Their parents stated that they and their children had never used pyrethroids or organophosphorus acids in their homes or for medical reasons. We analysed their spot urine samples for six metabolites of organophosphorus insecticides [dimethyl-phosphate (DMP), diethyl-phosphate (DEP), dimethyl-thiophosphate (DMTP), diethyl-thiophosphate (DETP), dimethyl-dithiophosphate (DMDTP) and diethyl-dithiophosphate (DEDTP)] and for four metabolites of pyrethroids [cis-3-(2,2-dibromo-vinyl)-2,2-dimethyl-cyclopropane carboxylic acid (Br2CA), cis-3-(2,2-dichloro-vinyl)-2,2-dimethyl-cyclopropane carboxylic acid (cis-Cl2-CA ), trans-3-(2,2-dichloro-vinyl)-2,2-dimethyl-cyclopropane carboxylic acid (trans-Cl2-CA) and 4-fluoro-3-phenoxy-benzoic acid (F-PBA)] using gas chromatographic methods with mass-selective detection. The limit of detection was 0.1-0.2 microg/l for pyrethroid metabolites and 1 microg/l for metabolites of organophosphorus acids; in DMP it was 5 microg/l. The 95th percentiles of the urinary metabolite concentrations were, in microgrammes per litre, DMP 158, DMTP 180, DMDTP 12, DEP 17, DETP 8, DEDTP <1; Br2CA 0.30, cis-Cl2-CA 0.44, trans-Cl2-CA 1.22, F-PBA 0.30. There were no correlations between urinary metabolite levels and the age of the children. Current background levels of internal exposure to pyrethroids and organophosphorus insecticides in children and adolescents in Germany are shown. Exposure to these substances in the general population is thought to occur mainly via residues in the diet. The level of background internal pyrethroid exposure in the children is orders of magnitude lower than the corresponding acceptable daily intake (ADI) values published, but the level of internal organophosphate exposure may reach and even exceed ADI values. This observation demands further investigation.


The impact of radiotherapy on swallowing and speech in patients who undergo total laryngectomy.
Quality of life studies have shown no detrimental effect with radiotherapy (RT) in patients who have a total laryngectomy. We wished to determine the effect of RT (initial or postoperative) specifically on the swallowing and voice function in patients treated by total laryngectomy (TL) for carcinoma of the larynx. Multicenter chart review. Multicenter study in the Greater Manchester and Lancashire area. A total of 121 postlaryngectomy patients all of whom had completed definitive treatment at least 6 months before this study. Twenty-six patients had total laryngectomy as a single modality treatment and 95 had total laryngectomy and radiotherapy. Swallowing (solid food, soft diet or fluid/PEG) and voice development. Swallowing was better in the group who had no radiotherapy (P = 0.0037). There was no difference in voice function between the two groups. We also demonstrated that females had a worse swallowing outcome (P = 0.0101), as did advanced nodal stage (P = 0.001). RT adversely affects the swallowing results but not the speech results after TL when given either as initial treatment or postoperatively. This should be kept in mind in the decision-making process in the treatment of patients with carcinoma of the larynx.


A man with a prosthetic ear and multiple pulmonary nodules.
Basal cell carcinoma is generally regarded as a relatively indolent tumor easily controlled with local therapy. When neglected or inadequately treated this tumor can become locally aggressive and in rare circumstances metastasize. This report documents a case of basal cell carcinoma metastatic to the lung that resulted in rapidly progressive respiratory failure and death.


Analysis of mammalian fibroblast transformation by normal and mutated human EGF receptors.
Activation of the EGF receptor (c-erbB) tyrosine kinase has been implicated in tumorigenesis, either by overexpression of the normal receptor in the presence of EGF, or through expression of a truncated receptor lacking the EGF binding domain as in the viral oncogene v-erbB. Here, normal and truncated human EGF receptors expressed in Rat1 fibroblasts were analysed for receptor tyrosine kinase activity and several transformation parameters in comparison with polyoma middle T and EJ-ras. Expression of a truncated EGF receptor lacking the extracellular ligand binding domain induced transformation of immortalized rodent fibroblasts and appears to activate the intrinsic tyrosine kinase. The transformed phenotype becomes enhanced by further truncation of the C-terminal domain containing the tyrosine autophosphorylation sites P1 and P2. Over expression of EGF receptors with an intact extracellular region in transfected Rat1 cells shows EGF dependent transformation, which is reduced by C-terminal truncation. Transformation is dependent on the cellular receptor concentration and can be selected as a stable phenotype. We conclude that expression of receptors with a truncated EGF-binding domain alone is sufficient to transform mammalian fibroblasts, in contrast to chick fibroblasts transformed by v-erbB where additional deletion of C-terminal receptor sequences appears to be an absolute requirement.


Plasmodium berghei PIMMS2 Promotes Ookinete Invasion of the Anopheles gambiae Mosquito Midgut.
Mosquito midgut stages of the malaria parasite present an attractive biological system to study host-parasite interactions and develop interventions to block disease transmission. Mosquito infection ensues upon oocyst development that follows ookinete invasion and traversal of the mosquito midgut epithelium. Here, we report the characterization of PIMMS2 (Plasmodium invasion of mosquito midgut screen candidate 2), a Plasmodium berghei protein with structural similarities to subtilisin-like proteins. PIMMS2 orthologs are present in the genomes of all plasmodia and are mapped between the subtilisin-encoding genes SUB1 and SUB3 P. berghei PIMMS2 is specifically expressed in zygotes and ookinetes and is localized on the ookinete surface. Loss of PIMMS2 function through gene disruption by homologous recombination leads to normal development of motile ookinetes that exhibit a severely impaired capacity to traverse the mosquito midgut and transform to oocysts. Genetic complementation of the disrupted locus with a mutated PIMMS2 allele reveals that amino acid residues corresponding to the putative subtilisin-like catalytic triad are important but not essential for protein function. Our data demonstrate that PIMMS2 is a novel ookinete-specific protein that promotes parasite traversal of the mosquito midgut epithelium and establishment of mosquito infection.


Lateral orbital wall anomaly in a patient with thyroid eye disease.
An isolated defect in the lateral orbital wall is an uncommon occurrence, although cases of orbital floor, roof, and medial wall spontaneous dehiscence have been described in the literature. Here, we describe a lateral orbital wall defect in a patient with thyroid eye disease - which may be related to bony remodeling or may represent a rare congenital anomaly. Computed tomography (CT) prior to decompression surgery revealed a defect in the left lateral orbital wall. At the time of orbital decompression, prolapse of buccal fat into the orbit was noted once the subperiosteal plane was entered along the lateral orbital wall in the area of this bony defect. Literature review revealed only a few previous reports of lateral wall abnormalities, including two involving the inferior orbital fissure; one other case was associated with Down syndrome, and other reports consist of sphenoid hypoplasia associated with neurofibromatosis.


Symmetry cues for matching mirrored objects.
The study explores cues for matching pairs of objects. These objects were arranged into mirror-symmetrical displays and the task was to judge whether a pre-specified 180 degree rotation around the X, Y or Z axis carries one object into the mirror object. For some rotations, the object's mirror symmetry (M) and, for other rotations, the object's point-symmetry (P) could serve as a cue. The matching results suggest that M is a better cue than P, say M > P. Various attempts are made to explain this effect. The most promising one focuses on simplest structural object representations as these capture M and not P. It is furthermore plausible that M captured by reference frames at high hierarchical representation levels, say M1, serves as a better cue than M captured at low levels, say M2. The prediction M1 > M2 > P merely applies to the open surface objects in the experiment. For the closed solid objects in the experiment the expectation is M1 = M2 = P. Both predictions roughly agree with the accuracy and reaction-time data. The results suggest the perceptual relevance of representation cues and, aditionally, that cues stemming from reference frames at higher hierarchical representation levels are more effective than those from lower levels.


[The nitrous oxide system under conditions of chronic cerebral dopamine deficiency and hypoxia].
Acute experiments were conducted on adult Wistar rats two months after unilateral microinjection of the 6-hydroxydopamine into the medial forebrain bundle. This led to a greater than 90% decrease of dopamine, fell by 70% of the content of the stable metabolite of nitric oxide-nitrite anion and by up to 80% of nitric oxide synthase activity in the lesioned neostriatum. Nitric oxide synthase activity fell by 30% in the heart, by 60% in the aorta, while nitrite anion decreased in the heart and aorta, in blood plasma and erythrocytes by 45%, 40%, 70%, 30% compared with controls rats, respectively. Lesioned rats also showed changes in the pattern of responses of the smooth muscle of the aorta predominantly to endothelium-dependent (acetylcholine), rather than to endothelium-independent (nitroprusside) vasoreactivity. The amplitude of acetylcholine-induced relaxation of aortic smooth muscle decreased 4-fold, the rate of this response decreased 5-fold and the latent period increased 4-fold. Exposure of lesioned rats, controls rats to moderate hypoxia for 30 min resulted in a rise of nitrite anion content in all tissues; it increased 2-fold in erythrocytes of lesioned rats vs controls rats, in the lesioned and unlesioned neostriatum hypoxia restored the values to controls, while in the heart, aorta and plasma the levels were also increased but did not reach control values. Hypoxia also led to a rise of nitric oxide synthase activity in both lesioned rats and controls rats. However, compared to normoxia, the levels increased over 2 times in the lesioned neostriatum and in the heart, whereas in controls the increase was less that 2-fold. Hypoxia resulted in partial normalization of the functional deficit in endothelium-dependent dilatory responses of aortic smooth muscle. We conclude that the disturbances in nitric oxide system induced by mesostriatal dopaminergic lesions in animals may have relevance to Parkinson's disease. The improvement with ambient hypoxia in quantitative and functional aspects of the disturbances in nitric oxide system may also have relevance in the management of the disease.


Interactions among pelvic organ protrusion, levator ani descent, and hiatal enlargement in women with and without prolapse.
Pelvic organ prolapse has 2 components: (1) protrusion of the pelvic organs beyond the hymen; and (2) descent of the levator ani. The Pelvic Organ Prolapse Quantification system measures the first component, however, there remains no standard measurement protocol for the second mechanism. We sought to test the hypotheses that: (1) difference in the protrusion area is greater than the area created by levator descent in prolapse patients compared with controls; and (2) prolapse is more strongly associated with levator hiatus compared to urogenital hiatus. Midsagittal magnetic resonance imaging scans from 30 controls, 30 anterior predominant, and 30 posterior predominant prolapse patients were assessed. Levator area was defined as the area above the levator ani and below the sacrococcygeal inferior pubic point line. Protrusion area was defined as the protruding vaginal walls below the levator area. The levator hiatus and urogenital hiatus were measured. Bivariate analysis and multiple comparisons were performed. Bivariate logistic regression was performed to assess prolapse as a function of levator hiatus, urogenital hiatus, levator area, and protrusion. Pearson correlation coefficients were calculated. The levator area for the anterior (34.0 ± 6.5 cm2) and posterior (35.7 ± 8.0 cm2) prolapse groups were larger during Valsalva compared to controls (20.9 ± 7.8 cm2, P < .0001 for both); similarly, protrusion areas for the anterior (14.3 ± 6.2 cm2) and posterior (14.4 ± 5.7 cm2) prolapse groups were both larger compared to controls (5.0 ± 1.8 cm2, P < .0001 for both). The levator hiatus length for the anterior (7.2 ± 1 cm) and posterior (6.9 ± 1 cm) prolapse groups were longer during Valsalva compared to controls (5.2 ± 1.5 cm, P < .0001 for both); similarly, urogenital hiatus lengths for the anterior (5.7 ± 1 cm) and posterior (6.3 ± 1.1 cm) prolapse groups were both longer than controls (3.8 ± 0.8 cm, P < .0001 for both). The difference in levator area in prolapse patients compared with controls was greater than the difference in protrusion area (14.0 ± 7.2 cm2 vs 9.4 ± 5.9 cm2, P < .0002). The urogenital hiatus was more strongly associated with prolapse than the levator hiatus (odds ratio, 12.9; 95% confidence interval, 4.1-39.2, and odds ratio, 4.3; 95% confidence interval, 2.3-7.5). Levator hiatus and urogenital hiatus are both correlated with levator and protrusion areas, and all were associated with maximum prolapse size (P ≤ .001, for all comparisons). In prolapse, the levator area increases more than the protrusion area and both the urogenital hiatus and levator hiatus are larger. The odds of prolapse for an increase in the urogenital hiatus are 3 times larger than for the levator hiatus, which leads us to reject both the original hypotheses.


A specific immune and lymphatic profile characterizes the pre-metastatic state of the sentinel lymph node in patients with early cervical cancer.
The lymph node (LN) pre-metastatic niche is faintly characterized in lymphophilic human neoplasia, although LN metastasis is considered as the strongest prognostic marker of patient survival. Due to its specific dissemination through a complex bilateral pelvic lymphatic system, early cervical cancer is a relevant candidate for investigating the early nodal metastatic process. In the present study, we analyzed in-depth both the lymphatic vasculature and the immune climate of pre-metastatic sentinel LN (SLN), in 48 cases of FIGO stage IB1 cervical neoplasms. An original digital image analysis methodology was used to objectively determine whole slide densities and spatial distributions of immunostained structures. We observed a marked increase in lymphatic vessel density (LVD) and a specific capsular and subcapsular distribution in pre-metastatic SLN when compared with non-sentinel counterparts. Such features persisted in the presence of nodal metastatic colonization. The inflammatory profile attested by CD8+, Foxp3, CD20 and PD-1expression was also significantly increased in pre-metastatic SLN. Remarkably, the densities of CD20+ B cells and PD-1 expressing germinal centers were positively correlated with LVD. All together, these data strongly support the existence of a pre-metastatic dialog between the primary tumor and the first nodal relay. Both lymphatic and immune responses contribute to the elaboration of a specific pre-metastatic microenvironment in human SLN. Moreover, this work provides evidence that, in the context of early cervical cancer, a pre-metastatic lymphangiogenesis occurs within the SLN (pre-metastatic niche) and is associated with a specific humoral immune response.


Investigating the Effect of Perinatal Nicotine Exposure on Dopaminergic Neurons in the VTA Using miRNA Expression Profiles.
Maternal smoking during pregnancy is associated with developmental, cognitive, and behavioral disorders, including low birth weight, attention deficit hyperactivity disorder, learning disabilities, and drug abuse later in life. Nicotine activates the reward-driven behavior characteristic of drug abuse. Dopaminergic (DA) neurons originating from the ventral tegmental area (VTA) of the brain, which are stimulated by nicotine and other stimuli, are widely implicated in the natural reward pathway that is known to contribute to addiction. In recent years, microRNAs have been implicated in disrupting regulatory mechanisms due to their capability of targeting multiple genes and thus inducing downstream effects along many pathways. In order to investigate miRNA expression of dopaminergic neurons from the VTA, we employed patch clamping to identify and harvest both DA and non-DA neurons from rats perinatally exposed to nicotine for use in single-cell RT-qPCR. Our data indicated that miR-140-5p and miR-140-3p were upregulated in DA neurons; while miR-140-3p and miR-212 were differentially expressed in non-DA neurons. A functional enrichment analysis was also performed on our miRNA-gene prediction network and predicted that our miRNAs target genes involved in drug response and neuroplasticity.


An escalating dose/multiple high-dose binge pattern of amphetamine administration results in differential changes in the extracellular dopamine response profiles in caudate-putamen and nucleus accumbens.
Amphetamine (AMPH)-induced psychosis is most frequently associated with a chronic high-dose "binge" or "run" pattern of stimulant abuse, generally preceded by a period of gradually escalating doses of the drug. We showed previously that animals subjected to such a regimen of AMPH administration developed, over multiple daily binges, a unique pattern of behavioral response that included a decrease in stereotypy and a pronounced increase in locomotion. Because of the involvement of mesolimbic and mesostriatal dopamine (DA) pathways in locomotion and stereotypy, respectively, we hypothesized that a persistent shift in the relative magnitude of caudate-putamen (CP) and nucleus accumbens (NAC) DA transmission may contribute to this altered behavioral profile. To test this hypothesis, we examined CP and NAC extracellular DA in response to multiple high-dose AMPH binges. Our results revealed that with multiple binges the CP DA response but not the NAC response developed a profound tolerance/tachyphylaxis to the drug-induced increase in extracellular transmitter. These differential regional response alterations seem to correspond to the shift in the relative expression of stereotypy and locomotion. We hypothesize that changes in DA synthesis, perhaps mediated by regionally specific adaptations in DA autoreceptor function, contribute to the differential extracellular transmitter response profiles, and suggest that these neurochemical changes may have important implications for the mechanisms underlying the addictive and psychotogenic properties of AMPH.


[Center specific duplex Doppler threshold values in carotid artery stenosis].
The traditional categorizations of color Doppler ultrasonography (CDUS) used for grading internal carotid artery stenosis are not consistent with the multicenter controlled randomized studies of carotid artery endarterectomy. The purpose of this study was to determine duplex Doppler threshold values specific to our center for classification of critical internal carotid artery stenosis which is determined according to the results obtained from multi-center clinical studies. CDUS and digital subtraction angiography examinations were performed in 147 patients who were considered suitable for carotid endarterectomy. Using the values of peak systolic velocity (PSV), end diastolic velocity and peak systolic velocity ratio (PSV ICA/CCA), the optimum duplex Doppler threshold values were determined for internal artery carotid stenoses by ROC (receiver operating characteristics) analysis. The most accurate threshold levels for detection of ICA stenoses were found to be a PSV 133 cm/s for 50-99% stenosis, PSV ICA/CCA 2.6 for 60-99% stenosis and PSV ICA/CCA 3.6 for 70-99% stenosis. Sensitivity, specificity, positive predictive value, negative predictive value and accuracy estimated using optimal duplex criteria for identification of ICA stenoses were 88%, 85% 78%, 91%, 86% for 50-69%, 94%, 87%, 71%, 98%, 89% for 60-99% stenosis, 92%, 92%, 65%, 99%, 92% for 70-99% stenosis, respectively. The duplex criteria established in our center have high accuracy. Every center should establish its own duplex criteria for the diagnosis of carotid artery stenosis.


Immune responses to zoster vaccines.
There are two licensed herpes zoster vaccines. One is a live vaccine (ZVL) based on an attenuated varicella-zoster virus (VZV). The other is a recombinant vaccine (RZV) based on the VZV glycoprotein E (gE) combined with AS01B, a multicomponent adjuvant system. RZV is superior to ZVL in efficacy, and differs from ZVL in that protection is not diminished by the age of the vaccinee and has not waned significantly during 4 years of follow-up. Immunologic studies demonstrated higher peak memory and persistence of T cell responses in RZV compared with ZVL recipients. RZV recipients also showed development and persistence of polyfunctional T cell responses. Taken together, we conclude that the immunologic data parallel and support the higher efficacy over time of RZV compared with ZVL.


Susceptibility of Manduca sexta to Cry1Ab toxin of Bacillus thuringiensis correlates directly to developmental expression of the cadherin receptor BT-R(1).
The cadherin receptor BT-R(1), localized in the midgut epithelium of the tobacco hornworm, Manduca sexta, is coupled to programmed oncotic-like cell death, which is triggered by the univalent binding of the Cry1Ab toxin of Bacillus thuringiensis (Bt) to the receptor. Kinetic analysis of BT-R(1) expression during larval development reveals that the density of BT-R(1) on the midgut surface increases dramatically along with an equivalent rise in the concentration of Cry1Ab toxin molecules needed to kill each of the five larval stages of the insect. The increase in the number of BT-R(1) molecules per midgut surface area requires additional toxin molecules to kill older versus younger larvae, as evidenced by the corresponding LC(50) values. Based on these observations, we developed a mathematical model to quantify both the expression of BT-R(1) and the susceptibility of M. sexta larvae to the Cry1Ab toxin. Interestingly, the toxin-receptor ratio remains constant during larval development regardless of larval size and mass. This ratio apparently is critical for insecticidal activity and the decrease in toxin effectiveness during larval development is due primarily to the number of effective toxins and available receptors in the larval midgut. Evidently, susceptibility of M. sexta to the Cry1Ab toxin of Bt correlates directly to the developmental expression of BT-R(1) in this insect.


Bulimia Symptoms in Russian Youth: Prevalence and Association With Internalizing Problems.
There has been limited research on bulimia symptoms in adolescents from the general population outside the United States. This study aimed to evaluate the prevalence of bulimia symptoms in Russian youth and explore the associations between a clinical level of self-reported probable bulimia nervosa (BN) and internalizing problems, binge drinking and functional impairment by gender. Data were collected from a representative sample of school students (N = 2,515, 59.5% female) from Northern Russia [age M (SD) = 14.89 ± 1.13 years]. Probable BN and internalizing psychopathology were assessed using self-report scales. Chi-square and independent sample t-tests were used to compare respondents' demographic characteristics and disordered eating behaviors. GLM multivariate analysis of covariance was used to assess the associations between probable BN, functional impairment and mental health problems (MHP) by gender. Analyses showed that the 3-month prevalence of probable BN was higher in girls (3.9%) than in boys (1.2%). Probable BN was associated with depressive and anxiety symptoms, somatic anxiety, somatic complaints, binge drinking and functional impairment. Boys reported a higher level of problem scores in relation to probable BN. Our findings suggest that bulimia symptoms are prevalent in Russian adolescents and are associated with MHP and functional impairment. Timely recognition of bulimia symptoms and associated MHP is important for early prevention and intervention strategies.


Targeting of chromatin readers: a novel strategy used by the Shigella flexneri virulence effector OspF to reprogram transcription.
Shigella flexneri, a gram-negative bacterium responsible of bacillary dysentery, uses multiple strategies to overcome host immune defense. We have decrypted how this bacterium manipulates host-cell chromatin binders to take control of immune gene expression. We found that OspF, an injected virulence factor previously identified as a repressor of immune gene expression, targets the chromatin reader HP1γ. Heterochromatin Protein 1 family members specifically recognize and bind histone H3 methylated at Lys 9. Although initially identified as chromatin-associated transcriptional silencers in heterochromatin, their location in euchromatin indicates an active role in gene expression. Notably, HP1γ phosphorylation at Serine 83 defines a subpopulation exclusively located to euchromatin, targeted to the site of transcriptional elongation. We showed that OspF directly interacts with HP1γ, and causes HP1 dephosphorylation, suggesting a model in which this virulence effector "uses" HP1 proteins as beacons to target and repress immune gene expression (Harouz, et al. EMBO J (2014)). OspF alters HP1γ phosphorylation mainly by inactivating the Erk-activated kinase MSK1, spotlighting it as a new HP1 kinase. In vivo, infectious stresses trigger HP1γ phosphorylation in the colon, principally in the lamina propria and the intestinal crypts. Several lines of evidence suggest that HP1 proteins are modified as extensively as histones, and decrypting the impact of these HP1 post-translational modifications on their transcriptional activities in vivo will be the next challenges to be taken up.


Effects of aging and HEMA content on the translucency, fluorescence, and opalescence properties of experimental HEMA-added glass ionomers.
Changes in the translucency, fluorescence, and opalescence of experimental 10-50% 2-hydroxyethyl methacrylate (HEMA)-added glass ionomers (HAGIs) after 5,000 cycles of thermocycling were determined and compared with those of commercial resin-modified glass ionomers (RMGIs). Changes in the translucency (TP), fluorescence (FL), and opalescence (OP) parameters were in the range of -3.5 to 0.2, -2.3 to 0.3 and -2.6 to 9.1 units respectively for HAGIs; and -0.9 to 0.3, -0.7 to 0.6, and 1.1 to 2.3 units respectively for RMGIs. Changes in the TP, FL, and OP of HAGIs were influenced by the HEMA content and powder shade, and were generally larger than those of RMGIs. Since the changes in TP, FL, and OP of experimental HAGIs were influenced by the HEMA content, there arises a need to determine the optimal HEMA ratio to attain high stability for these optical properties. In addition, results of this study showed that apart from optimal HEMA ratio, future studies should include other aspects and factors that contribute to age-dependent changes in optical properties.


Utilization of Transcriptome, Small RNA, and Degradome Sequencing to Provide Insights Into Drought Stress and Rewatering Treatment in Medicago ruthenica.
Drought is a major limiting factor in foraging grass yield and quality. Medicago ruthenica (M. ruthenica) is a high-quality forage legume with drought resistance, cold tolerance, and strong adaptability. In this study, we integrated transcriptome, small RNA, and degradome sequencing in identifying drought response genes, microRNAs (miRNAs), and key miRNA-target pairs in M. ruthenica under drought and rewatering treatment conditions. A total of 3,905 genes and 50 miRNAs (45 conserved and 5 novel miRNAs) were significantly differentially expressed in three test conditions (CK: control, DS: plants under drought stress, and RW: plants rewatering after drought stress). The degradome sequencing (AllenScore < 4) analysis revealed that 104 miRNAs (11 novel and 93 conserved miRNAs) were identified with 263 target transcripts, forming 296 miRNA-target pairs in three libraries. There were 38 differentially expressed targets from 16 miRNAs in DS vs. CK, 31 from 11 miRNAs in DS vs. RW, and 6 from 3 miRNAs in RW vs. CK; 21, 18, and 3 miRNA-target gene pairs showed reverse expression patterns in DS vs. CK, DS vs. RW, and RW vs. CK comparison groups, respectively. These findings provide valuable information for further functional characterization of genes and miRNAs in response to abiotic stress, in general, and drought stress in M. ruthenica, and potentially contribute to drought resistance breeding of forage in the future.


An unusual dentigerous cyst in a young child.
A dentigerous cyst is an odontogenic developmental cyst associated with the crown of an unerupted tooth. This article describes the case of a 7-year-old girl with painful swelling in the left facial region, prolonged retention of tooth No. 9d, and a displaced tooth No. 9 in eruption. Radiographic examination revealed an extensive radiolucent lesion of approximately 28 mm and displacement of teeth No. 10 and 11. Marsupialization was performed, teeth No. 8d and 9d were removed, and the swelling reduced immediately. At a 2-year follow-up, radiographs revealed that teeth No. 10 and 11 had erupted but were displaced. Routine radiographic examinations are important for children during the eruption phase to prevent and detect any alterations that could damage the permanent dentition and occlusion.


Selectivity of BI 689648, a Novel, Highly Selective Aldosterone Synthase Inhibitor: Comparison with FAD286 and LCI699 in Nonhuman Primates.
The mineralocorticoid aldosterone is an important regulator of blood pressure, volume, and electrolyte balance. However, excess aldosterone can be deleterious as a driver of vascular remodeling and tissue fibrosis associated with cardiometabolic diseases. Aldosterone synthase (AS) inhibitors (ASI) attenuate the production of aldosterone directly and have been proposed as an alternative to mineralocorticoid receptor antagonists for blocking the pathologic effects of excess aldosterone. Discovery of selective ASIs has been challenging because of the high sequence identity (93%) AS shares with cortisol synthase (CS), and the low identity of rodent AS compared with human (63%). Using cynomolgus (cyno) monkey-based models, we identified BI 689648 [6-(5-methoxymethyl-pyridin-3-yl)-3,4-dihydro-2H-[1,8]naphthyridine-1-carboxylic acid amide], a novel, highly selective ASI that exhibits an in vitro IC50 of 2 nM against AS and 300 nm against CS (150-fold selectivity) compared with the recently described ASIs FAD286 [4-(5,6,7,8-tetrahydroimidazo[1,5-a]pyridin-5-yl)benzonitrile] (3 nM AS; 90 nM CS; 40-fold) and LCI699 (4-[(5R)-6,7-dihydro-5H-pyrrolo[1,2-c]imidazol-5-yl]-3-fluorobenzonitrile) (10 nM AS; 80 nM CS; 8-fold). After oral administration in cyno monkeys, BI 689648 (5 mg/kg) exhibits a peak plasma concentration of ∼500 nM. For in vivo profiling we used an adrenocorticotropin-challenge model in which BI 689648 was >20-fold more selective compared with FAD286 and LCI699. Because both FAD286 and LCI699 failed to provide adequate selectivity for CS when tested in patients, the desire for more selective molecules to test the ASI hypothesis remains high. Therefore, highly selective aldosterone synthase inhibitors such as BI 689648 represent an important step forward toward developing ASIs with greater potential for clinical success in cardiometabolic diseases.


Splayed polarization in the ferroelectric phase of a bent-core liquid crystal as studied by optical second-harmonic generation.
Splayed polarization orientation has been confirmed in the anticlinic ferroelectric phase of a bent-core liquid crystal containing chiral end chains, P-8 -OPIMB6(*), by optical second-harmonic generation (SHG). The in-plane anisotropy of the SHG signal exhibits an unusual six-peak pattern under the rotation of parallel polarizers in the absence of an electric field, while a usual four-peak pattern simulatable using a uniform molecular orientation model is observed under a field. The unusual pattern was explained by considering a splayed polarization orientation including the effect of polarization charges.


Root canal retreatment.
Root canal therapy is not always successful and an increasing number of patients are requesting retreatment to address intraradicular infection. The armamentarium available to assist the dentist, some of which is described in this article, has never been greater.


Termination of macroreentrant atrial arrhythmias by pacing stimuli without global propagation.
Electrical stimulation during ventricular tachycardia resulting in tachycardia termination without global propagation (TWGP) is a well-recognized phenomenon. However, there is a paucity of literature showing a similar phenomenon in atrial arrhythmias. The purpose of the study was to evaluate the significance of TWGP in atrial arrhythmias. Electrophysiological studies performed from 2000 to 2019 at Methodist Hospital, Indiana University were reviewed retrospectively. Thirty-four patients were identified in whom stimulation during atrial tachycardia/flutter resulted in TWGP. Of the 34 patients, 12 (29%) had cavotricuspid isthmus (CTI)-dependent atrial flutter and 22 (71%) had other atrial arrhythmias during which TWGP was seen. Mean age of the population was 53 ± 13 years; and 68% were male. Previous catheter ablation for atrial fibrillation, atrial flutter, or other atrial tachyarrhythmias had been performed in 70.5%, and 44% previously had undergone cardiac surgery involving the atria. Congenital heart disease was present in 20.5%; 3 patients were status post lung transplant. Mean cycle length of atrial arrhythmia in which TWGP was seen was 317 ± 76 ms. The sites at which TWGP was seen reproducibly were highly specific for successful termination of the arrhythmias with radiofrequency energy. The arrhythmia circuits were 12 CTI-dependent atrial flutter, 11 left atrial macroreentrant atrial tachycardia (MRAT), 1 involving both left and right atria, and 8 were other right atrial MRAT. Termination of macroreentrant atrial arrhythmias by pacing stimuli without global propagation identifies a narrow diastolic isthmus at which catheter ablation is highly effective.


Neurotrophic effects of extracellular guanosine.
Central nervous system (CNS) astrocytes release guanosine extracellularly, that exerts trophic effects. In CNS, extracellular guanosine (GUO) stimulates mitosis, synthesis of trophic factors, and cell differentiation, including neuritogenesis, is neuroprotective, and reduces apoptosis due to several stimuli. Specific receptor-like binding sites for eGUO in the nervous system may mediate its effects through both MAP kinase and PI3-kinase signalling pathways. Extracellular guanine (eGUA) also exerts several effects; the trophic effects of eGUO are likely regulated by conversion of eGUO to eGUA by a membrane located purine nucleoside phosphorylase (ecto-PNP) and by conversion of eGUA to xanthine by guanine deaminase.


[Intra-anesthetic arterial hypotension in elderly patients during emergency surgery: what are the risk factors?]
Emergency anesthesia in elderly patients aged 65 years and older is complex. The occurrence of intraoperative incidents and arterial hypotension is conditioned by patients' initial health status and by the quality of intraoperative management. This study aimed to determine the incidence of intra-anesthetic arterial hypotension in elderly patients during emergency surgery and to assess the involvement of certain factors in its occurrence: age, sex, patient's history, ASA class, anesthetic technique. We conducted a retrospective descriptive and analytical study in the Emergency Surgery Department at the Aristide Le Dantec University Hospital from 1 March 2014 to 28 February 2015. We collected data from 210 patients out of 224 elderly patients aged 65 years and older undergoing emergency anesthesias (10.93%). Data of 101 men and 109 women were included in the analysis, of whom 64.3% had at least one defect. Patients' preoperative status was assessed using American Society of Anesthesiology (ASA) classification: 71% of patients were ASA class 1 and 2 and 29% were ASA class 3 and 4. Locoregional anesthesia was the most practiced anesthetic technique (56.7%). 28 patients (13.33%) had intra-anesthetic arterial hypotension, of whom 16 under general anesthesia and 12 under locoregional anesthesia. It was more frequent in patients with high ASA class and a little less frequent in patients with PAH and underlying heart disease. Arterial hypotension in elderly patients during emergency surgery exposes the subject to the risk of not negligible intraoperative hypotension, especially in patients with high ASA class. Prevention is based on adequate preoperative assessment and anesthetic management.


The nitric oxide donor LA 419 decreases ischemic brain damage.
Stroke represents a major clinical problem with limited available therapeutic treatments. Nitric oxide (NO) and the enzymes that produce it are involved in the pathogenesis of this disease. Here we investigated whether the novel NO donor LA 419 was able to ameliorate the consequences of stroke in an experimental model of global ischemia. We observed a sharp increase in the amounts of inducible NO synthase (iNOS) and nitrotyrosine in the cerebral cortex of experimental rats and a moderate increase of neuronal NO synthase (nNOS), as demonstrated by immunohistochemistry, Western blotting, and enzymatic activity assays. Treatment of these animals with LA 419 completely prevented ischemia-induced upregulation of nitrergic markers. Magnetic resonance imaging of the experimental brains showed a marked decrease in apparent diffusion coefficient (ADC) following ischemia-reperfusion, which was significantly corrected by pre-treatment with LA 419. These results clearly show that LA 419 is an efficient modulator of NO-related pathophysiological events and could eventually be used for the treatment of patients with cerebrovascular pathologies.


Illinois Dental Sealant Program: 1986-1996.
Dental sealants have been shown to be an effective means of preventing pit and fissure caries. Despite more than 20 years of research and overwhelming evidence in support of the use of sealants, the dental profession has been slow to make use of them. This article explores the strategy used by the Illinois Dental Sealant Program to increase dental sealant use.


Antibody-mediated elimination of malaria parasites (plasmodium berghei) in vivo.
An infective preparation of extracellular blood forms (FP) of Plasmodium berghei was used to study some aspects of the interaction between protective antibodies and malaria parasites. FP but not infected erythrocytes (IRBC) were shown by the fluorescent antibody technique to be coated by antibodies after in vitro incubation with immune serum. Preincubation of both FP and IRBC with immune serum followed by their washing did not result in enhanced elimination of the parasites in vivo. However, FP preincubated with immune serum and subsequently washed were eliminated more efficiently than FP preincubated with normal serum if the preparations were injected with some immune serum. Such an increase in the efficiency of elimination was not detected with similarly pretreated IRBC. It is thus probable that protective antibodies acted in vivo against extracellular parasites rather than against parasites in erythrocytes. The interaction between parasites and antibodies may be of a highly reversible nature, and washing of the in vitro-treated parasites may cause elution of antibody from the sensitized parasites so that the amount of antibody on the parasite falls below the critical level required for in vivo elimination.