Regulation of thyroid hormone-, oestrogen- and androgen-related genes by triiodothyronine in the brain of Silurana tropicalis.
Amphibian metamorphosis is an excellent example of hormone-dependent control of development. Thyroid hormones (THs) regulate almost all aspects of metamorphosis, including brain development and larval neuroendocrine function. Sex steroids are also important for early brain function, although little is known about interactions between the two hormonal systems. In the present study, we established brain developmental profiles for thyroid hormone receptors (tralpha and trbeta), deiodinases (dio1, dio2 and dio3), aromatase (cyp19) mRNA and activity, oestrogen receptors (eralpha and erbeta), androgen receptor (ar) and 5alpha-reductases (srd5alpha1 and srd5alpha2) mRNA during Silurana (Xenopus) tropicalis metamorphosis. Real-time reverse transcriptase-polymerase chain reaction analyses revealed that all of the genes were expressed in the brain and for most of the genes expression increased during development, with the exception of dio2, srd5alpha1 and srd5alpha2. The ability of premetamorphic tadpoles to respond to exogenous THs was used to investigate the regulation of TH- and sex steroid-related genes in the brain during development. Exposure of premetamorphic tadpoles to triiodothyronine (T3; 0, 0.5, 5 and 50 nm) for 48 h resulted in concentration-dependent increases in trbeta, dio2, dio3, eralpha and erbeta. Expression of srd5alpha2 showed large increases (six- to 7.5-fold) for all three concentrations of T3. No changes were detected in dio1, ar and cyp19 transcript levels; however, cyp19 activity increased significantly at 50 nm T3. The results obtained suggest that expression of TH-related genes and er during development could be regulated by rising levels of THs, as previously documented in Lithobates (Rana) pipiens. The positive regulation of srd5alpha by T3 in the brain suggests that endogenous TH levels help maintain or control the rate at which srd5alpha mRNA levels decrease as metamorphosis progresses. Finally, we have identified sex steroid-related genes that are responsive to T3, providing additional evidence of crosstalk between THs and sex steroids in the tadpole brain.


Identifying adult asthma phenotypes using a clustering approach.
There is a need to improve asthma characterisation by integrating multiple aspects of the disease. The aim of the present study was to identify distinct asthma phenotypes by applying latent class analysis (LCA), a model-based clustering method, to two large epidemiological studies. Adults with asthma who participated in the follow-up of the Epidemiological Study on the Genetics and Environment of Asthma (EGEA2) (n = 641) and the European Community Respiratory Health Survey (ECRHSII) (n = 1,895) were included. 19 variables covering personal characteristics, asthma symptoms, exacerbations and treatment, age of asthma onset, allergic characteristics, lung function and airway hyperresponsiveness were considered in the LCA. Four asthma phenotypes were distinguished by the LCA in each sample. Two phenotypes were similar in EGEA2 and ECRHSII: active treated allergic childhood-onset asthma and active treated adult-onset asthma. The other two phenotypes were composed of subjects with inactive or mild untreated asthma, who differed by atopy status and age of asthma onset (childhood or adulthood). The phenotypes clearly discriminated populations in terms of quality of life, and blood eosinophil and neutrophil counts. The LCAs revealed four distinct asthma phenotypes in each sample. Considering these more homogeneous phenotypes in future studies may lead to a better identification of risk factors for asthma.


Methotrexate in Psoriasis and Psoriatic Arthritis.
Methotrexate (MTX) is the most commonly prescribed first-line therapy in psoriatic arthritis (PsA) internationally and is also commonly used in the treatment of psoriasis. However, data supporting its use in PsA are limited and significant toxicities can occur. This article summarizes a debate at the 2019 Group for Research and Assessment of Psoriasis and Psoriatic Arthritis (GRAPPA) annual meeting that focused on the use of MTX in psoriasis and PsA. Four clinicians and 1 patient research partner presented clinical study data and the patient experience summarizing the efficacy, tolerability, and toxicity of MTX for both skin and musculoskeletal manifestations. A survey of attending GRAPPA members collected data on current and planned future use of MTX across the world.


Hospitalization for variceal hemorrhage in an era with more prevalent cirrhosis.
To examine hospitalization rates for variceal hemorrhage and relation to cause of cirrhosis during an era of increased cirrhosis prevalence. We performed a retrospective review of patients with cirrhosis and gastroesophageal variceal hemorrhage who were admitted to a tertiary care referral center from 1998 to 2009. Subjects were classified according to the etiology of their liver disease: alcoholic cirrhosis and non-alcoholic cirrhosis. Rates of hospitalization for variceal bleeding were determined. Data were also collected on total hospital admissions per year and cirrhosis-related admissions per year over the same time period. These data were then compared and analyzed for trends in admission rates. Hospitalizations for cirrhosis significantly increased from 611 per 100000 admissions in 1998-2001 to 1232 per 100000 admissions in 2006-9 (P value for trend < 0.0001). This increase was seen in admissions for both alcoholic and non-alcoholic cirrhosis (P values for trend < 0.001 and < 0.0001 respectively). During the same time period, there were 243 admissions for gastroesophageal variceal bleeding (68% male, mean age 54.3 years, 62% alcoholic cirrhosis). Hospitalizations for gastroesophageal variceal bleeding significantly decreased from 96.6 per 100000 admissions for the time period 1998-2001 to 70.6 per 100000 admissions for the time period 2006-2009 (P value for trend = 0.01). There were significant reductions in variceal hemorrhage from non-alcoholic cirrhosis (41.6 per 100000 admissions in 1998-2001 to 19.7 per 100000 admissions in 2006-2009, P value for trend = 0.007). Hospitalizations for variceal hemorrhage have decreased, most notably in patients with non-alcoholic cirrhosis, and this may reflect broader use of strategies to prevent bleeding.


Symbiosis-related pea genes modulate fungal and plant gene expression during the arbuscule stage of mycorrhiza with Glomus intraradices.
The arbuscular mycorrhiza association results from a successful interaction between genomes of the plant and fungal symbiotic partners. In this study, we analyzed the effect of inactivation of late-stage symbiosis-related pea genes on symbiosis-associated fungal and plant molecular responses in order to gain insight into their role in the functional mycorrhizal association. The expression of a subset of ten fungal and eight plant genes, previously reported to be activated during mycorrhiza development, was compared in Glomus intraradices-inoculated wild-type and isogenic genotypes of pea mutated for the PsSym36, PsSym33, and PsSym40 genes where arbuscule formation is inhibited or fungal turnover modulated, respectively. Microdissection was used to corroborate arbuscule-related fungal gene expression. Molecular responses varied between pea genotypes and with fungal development. Most of the fungal genes were downregulated when arbuscule formation was defective, and several were upregulated with more rapid fungal development. Some of the plant genes were also affected by inactivation of the PsSym36, PsSym33, and PsSym40 loci, but in a more time-dependent way during root colonization by G. intraradices. Results indicate a role of the late-stage symbiosis-related pea genes not only in mycorrhiza development but also in the symbiotic functioning of arbuscule-containing cells.


Helicobacter pylori infection and other bacteria in pancreatic cancer and autoimmune pancreatitis.
Helicobacter pylori (H. pylori) is an infectious agent influencing as much as 50% of the world's population. It is the causative agent for several diseases, most especially gastric and duodenal peptic ulcer, gastric adenocarcinoma and mucosa-associated lymphoid tissue lymphoma of the stomach. A number of other, extragastric manifestations also are associated with H. pylori infection. These include neurological disorders, such as Alzheimer's disease, demyelinating multiple sclerosis and Parkinson's disease. There is also evidence for a relationship between H. pylori infection and such dermatological diseases as psoriasis and rosacea as well as a connection with infection and open-angle glaucoma. Generally little is known about the relationship between H. pylori infection and diseases of the pancreas. Most evidence about H. pylori and its potential role in the development of pancreatic diseases concerns pancreatic adenocarcinoma and autoimmune forms of chronic pancreatitis. There is data (albeit not fully consistent) indicating modestly increased pancreatic cancer risk in H. pylori-positive patients. The pathogenetic mechanism of this increase is not yet fully elucidated, but several theories have been proposed. Reduction of antral D-cells in H. pylori-positive patients causes a suppression of somatostatin secretion that, in turn, stimulates increased secretin secretion. That stimulates pancreatic growth and thus increases the risk of carcinogenesis. Alternatively, H. pylori, as a part of microbiome dysbiosis and the so-called oncobiome, is proven to be associated with pancreatic adenocarcinoma development via the promotion of cellular proliferation. The role of H. pylori in the inflammation characteristic of autoimmune pancreatitis seems to be explained by a mechanism of molecular mimicry among several proteins (mostly enzymes) of H. pylori and pancreatic tissue. Patients with autoimmune pancreatitis often show positivity for antibodies against H. pylori proteins. H. pylori, as a part of microbiome dysbiosis, also is viewed as a potential trigger of autoimmune inflammation of the pancreas. It is precisely these relationships (and associated equivocal conclusions) that constitute a center of attention among pancreatologists, immunologists and pathologists. In order to obtain clear and valid results, more studies on sufficiently large cohorts of patients are needed. The topic is itself sufficiently significant to draw the interest of clinicians and inspire further systematic research. Next-generation sequencing could play an important role in investigating the microbiome as a potential diagnostic and prognostic biomarker for pancreatic cancer.


Intravascular mechanoreceptor modulation of renal sympathetic nerve activity in the cat.
Experiments were performed in chloralose-anesthetized cats to characterize intravascular mechanoreceptro input to renal nerve activity in the intact and vagotomized sinoaortic-denervated states. High-pressure intravascular mechanoreceptors were stimulated by rises in arterial pressure caused by norepinephrine. Low-pressure intravascular mechanoreceptors were stimulated by progressive blood volume expansion (14-23%) at a rate of 4.4 or 17.6 ml/min. In addition, veratrine was used to stimulate directly both high- and low-pressure receptors. In the intact animal the administration of norepinephrine or blood volume expansion was associated with substantial decreases in renal nerve activity. Veratrine also caused a large dose-related decrease in renal nerve activity. However, in the vagotomized sinoaortic-denervated animal there was no change in renal nerve activity with norepinephrine, volume expansion, or veratrine administration. These experiments demonstrate that the major afferent pathways for renal sympathetic circulatory reflexes are confined to the carotid sinus and aortic and vagus nerves. No evidence was found for a significant contribution from sympathetic afferent nerves.


Monoclonal antibodies for immunocytochemistry of progesterone receptors (PR) in various laboratory rodents, livestock, humans, and chickens: identification of two epitopes conserved in PR of all these species.
Over 90 mouse monoclonal antibodies have been raised against rabbit and human uterine progesterone receptor (PR). These antibodies, because of their specificity, are powerful tools with which to examine the localization, structure, and function of PR. A selection of 22 well characterized mABs was made to test their ability to give the best immunocytochemical staining of PR in various species. Comparative analysis of the antibodies led to the following conclusions. Li 417 (and, to a lesser extent, Let 126) was the best monoclonal in humans; Let 126 and Mi 60 were the most sensitive monoclonals in guinea pigs, rabbits, and monkeys. In sheep, sows, cows and mares as well as in rats and chickens Let 81 or Let 548 gave the best results (Let 126 was also effective in sows and mares, while Li 169 was also effective in sheep and cows). Two antibodies (Li 169 and Let 548) cross-reacted with PR in all of the species tested, including mammals and birds, and appeared to recognize two highly conserved antigenic sites. Remarkably, these conserved sequences are located in the highly variable N-terminal part of the receptor; they may, thus, be related to the still poorly understood function of this domain of the receptor.


Arrestin-3 binds c-Jun N-terminal kinase 1 (JNK1) and JNK2 and facilitates the activation of these ubiquitous JNK isoforms in cells via scaffolding.
Non-visual arrestins scaffold mitogen-activated protein kinase (MAPK) cascades. The c-Jun N-terminal kinases (JNKs) are members of MAPK family. Arrestin-3 has been shown to enhance the activation of JNK3, which is expressed mainly in neurons, heart, and testes, in contrast to ubiquitous JNK1 and JNK2. Although all JNKs are activated by MKK4 and MKK7, both of which bind arrestin-3, the ability of arrestin-3 to facilitate the activation of JNK1 and JNK2 has never been reported. Using purified proteins we found that arrestin-3 directly binds JNK1α1 and JNK2α2, interacting with the latter comparably to JNK3α2. Phosphorylation of purified JNK1α1 and JNK2α2 by MKK4 or MKK7 is increased by arrestin-3. Endogenous arrestin-3 interacted with endogenous JNK1/2 in different cell types. Arrestin-3 also enhanced phosphorylation of endogenous JNK1/2 in intact cells upon expression of upstream kinases ASK1, MKK4, or MKK7. We observed a biphasic effect of arrestin-3 concentrations on phosphorylation of JNK1α1 and JNK2α2 both in vitro and in vivo. Thus, arrestin-3 acts as a scaffold, facilitating JNK1α1 and JNK2α2 phosphorylation by MKK4 and MKK7 via bringing JNKs and their activators together. The data suggest that arrestin-3 modulates the activity of ubiquitous JNK1 and JNK2 in non-neuronal cells, impacting the signaling pathway that regulates their proliferation and survival.


Expectations and Experiences of Follow-up and Self-Care After Living Kidney Donation: A Focus Group Study.
Ensuring donor wellbeing warrants ongoing monitoring after living kidney donation. However, there is considerable variability in donor follow-up processes, including information provided to donors regarding self-care. Loss to follow-up is common, suggesting that the aims and benefits of monitoring and follow-up may not be apparent. We aimed to describe the experiences and expectations of living kidney donors regarding follow-up and self-care after donation. Participants from 3 transplant centers in Australia and Canada participated in 14 focus groups (n = 123). Transcripts were analyzed thematically. We identified 4 themes: lacking identification as a patient (invincibility and confidence in health, immediate return to normality, avoid burdening specialty services, redundancy of specialist attention, unnecessary travel), empowerment for health (self-preservation for devastating consequences, self-advocacy and education, needing lifestyle advice, tracking own results), safety net and reassurance (availability of psychosocial support, confidence in kidney-focused care, continuity and rapport, and access to waitlist priority), and neglect and inattention (unrecognized ongoing debilitations, primary focus on recipient, hospital abandonment, overlooking individual priorities, disconnected from system, coping with dual roles, and lacking support for financial consequences). Living kidney donors who felt well and confident about their health regarded specialist follow-up as largely unnecessary. However, some felt they did not receive adequate medical attention, were prematurely detached from the health system, or held unresolved anxieties about the consequences of their decision to donate. Ongoing access to healthcare, psychosocial support, and education may reassure donors that any risks to their health are minimized.


IFN-alpha and IL-10 induce the differentiation of human type 1 T regulatory cells.
CD4(+) T regulatory type 1 (Tr1) cells suppress Ag-specific immune responses in vitro and in vivo. Although IL-10 is critical for the differentiation of Tr1 cells, the effects of other cytokines on differentiation of naive T cells into Tr1 cells have not been investigated. Here we demonstrate that endogenous or exogenous IL-10 in combination with IFN-alpha, but not TGF-beta, induces naive CD4(+) T cells derived from cord blood to differentiate into Tr1 cells: IL-10(+)IFN-gamma(+)IL-2(-/low)IL-4(-). Naive CD4(+) T cells derived from peripheral blood require both exogenous IL-10 and IFN-alpha for Tr1 cell differentiation. The proliferative responses of the Tr1-containing lymphocyte populations, following activation with anti-CD3 and anti-CD28 mAbs, were reduced. Similarly, cultures containing Tr1 cells displayed reduced responses to alloantigens via a mechanism that was partially mediated by IL-10 and TGF-beta. More importantly, Tr1-containing populations strongly suppressed responses of naive T cells to alloantigens. Collectively, these results show that IFN-alpha strongly enhances IL-10-induced differentiation of functional Tr1 cells, which represents a first major step in establishing specific culture conditions to generate T regulatory cells for biological and biochemical analysis, and for cellular therapy to induce peripheral tolerance in humans.


Quantitative magnetic resonance (MR) neurography for evaluation of peripheral nerves and plexus injuries.
Traumatic conditions of peripheral nerves and plexus have been classically evaluated by morphological imaging techniques and electrophysiological tests. New magnetic resonance imaging (MRI) studies based on 3D fat-suppressed techniques are providing high accuracy for peripheral nerve injury evaluation from a qualitative point of view. However, these techniques do not provide quantitative information. Diffusion weighted imaging (DWI) and diffusion tensor imaging (DTI) are functional MRI techniques that are able to evaluate and quantify the movement of water molecules within different biological structures. These techniques have been successfully applied in other anatomical areas, especially in the assessment of central nervous system, and now are being imported, with promising results for peripheral nerve and plexus evaluation. DWI and DTI allow performing a qualitative and quantitative peripheral nerve analysis, providing valuable pathophysiological information about functional integrity of these structures. In the field of trauma and peripheral nerve or plexus injury, several derived parameters from DWI and DTI studies such as apparent diffusion coefficient (ADC) or fractional anisotropy (FA) among others, can be used as potential biomarkers of neural damage providing information about fiber organization, axonal flow or myelin integrity. A proper knowledge of physical basis of these techniques and their limitations is important for an optimal interpretation of the imaging findings and derived data. In this paper, a comprehensive review of the potential applications of DWI and DTI neurographic studies is performed with a focus on traumatic conditions, including main nerve entrapment syndromes in both peripheral nerves and brachial or lumbar plexus.


Cyclin B1/Cdk1 phosphorylation of mitochondrial p53 induces anti-apoptotic response.
The pro-apoptotic function of p53 has been well defined in preventing genomic instability and cell transformation. However, the intriguing fact that p53 contributes to a pro-survival advantage of tumor cells under DNA damage conditions raises a critical question in radiation therapy for the 50% human cancers with intact p53 function. Herein, we reveal an anti-apoptotic role of mitochondrial p53 regulated by the cell cycle complex cyclin B1/Cdk1 in irradiated human colon cancer HCT116 cells with p53(+/+) status. Steady-state levels of p53 and cyclin B1/Cdk1 were identified in the mitochondria of many human and mouse cells, and their mitochondrial influx was significantly enhanced by radiation. The mitochondrial kinase activity of cyclin B1/Cdk1 was found to specifically phosphorylate p53 at Ser-315 residue, leading to enhanced mitochondrial ATP production and reduced mitochondrial apoptosis. The improved mitochondrial function can be blocked by transfection of mutant p53 Ser-315-Ala, or by siRNA knockdown of cyclin B1 and Cdk1 genes. Enforced translocation of cyclin B1 and Cdk1 into mitochondria with a mitochondrial-targeting-peptide increased levels of Ser-315 phosphorylation on mitochondrial p53, improved ATP production and decreased apoptosis by sequestering p53 from binding to Bcl-2 and Bcl-xL. Furthermore, reconstitution of wild-type p53 in p53-deficient HCT116 p53(-/-) cells resulted in an increased mitochondrial ATP production and suppression of apoptosis. Such phenomena were absent in the p53-deficient HCT116 p53(-/-) cells reconstituted with the mutant p53. These results demonstrate a unique anti-apoptotic function of mitochondrial p53 regulated by cyclin B1/Cdk1-mediated Ser-315 phosphorylation in p53-wild-type tumor cells, which may provide insights for improving the efficacy of anti-cancer therapy, especially for tumors that retain p53.


Chemotherapy of disseminated germ cell tumors.
The treatment of patients with germ cell neoplasms has improved dramatically during the last 10 years. Combination chemotherapy with cisplatin + vinblastine + bleomycin (PVB) was initiated at Indiana University in 1974. In this original study 28 of 47 patients (60%) with disseminated testicular tumors are currently alive, with a minimal follow-up of 10 years. Our next PVB study demonstrated that the vinblastine dosage could be reduced 25% (0.4 mg/kg to 0.3 mg/kg), thereby significantly reducing toxicity, without compromising the cure rate. Our third generation study confirmed the fact that maintenance therapy was not necessary, and that optimal cure rates could be achieved with merely 12 weeks of PVB induction. Overall, with follow-up of 5 to 10 years, 202 of 272 patients (74%) with disseminated testicular cancer currently are alive on these three PVB protocols. Because of the success of salvage chemotherapy, demonstrating that approximately 25% of patients not curable by PVB achieved durable complete remissions (CRs) with cisplatin + VP-16 combination chemotherapy, our next study compared PVB to cisplatin + VP-16 + bleomycin (PVP16B) as initial chemotherapy for disseminated testicular cancer. This study demonstrated that the two induction regimens produced equivalent CR rates, but that PVP16B was the preferable induction regimen because of a statistically significant reduction in neuromuscular toxicity. Our current studies will evaluate whether three courses (9 weeks) of PVP16B compared to the standard four courses of PVP16B (12 weeks) will achieve similar cure rates in minimal moderate disseminated disease. In advanced disease presentations, we are comparing standard PVP16B to double-dose cisplatin (40 mg/M2 X 5) + VP-16 + bleomycin.


Imaging of mRNA-protein interactions in live cells using novel mCherry trimolecular fluorescence complementation systems.
Live cell imaging of mRNA-protein interactions makes it possible to study posttranscriptional processes of cellular and viral gene expression under physiological conditions. In this study, red color mCherry-based trimolecular fluorescence complementation (TriFC) systems were constructed as new tools for visualizing mRNA-protein interaction in living cells using split mCherry fragments and HIV REV-RRE and TAT-TAR peptide-RNA interaction pairs. The new mCherry TriFC systems were successfully used to image RNA-protein interactions such as that between influenza viral protein NS1 and the 5' UTR of influenza viral mRNAs NS, M, and NP. Upon combination of an mCherry TriFC system with a Venus TriFC system, multiple mRNA-protein interactions could be detected simultaneously in the same cells. Then, the new mCherry TriFC system was used for imaging of interactions between influenza A virus mRNAs and some of adapter proteins in cellular TAP nuclear export pathway in live cells. Adapter proteins Aly and UAP56 were found to associate with three kinds of viral mRNAs. Another adapter protein, splicing factor 9G8, only interacted with intron-containing spliced M2 mRNA. Co-immunoprecipitation assays with influenza A virus-infected cells confirmed these interactions. This study provides long-wavelength-spectrum TriFC systems as new tools for visualizing RNA-protein interactions in live cells and help to understand the nuclear export mechanism of influenza A viral mRNAs.


Determination of d-serine and related neuroactive amino acids in human plasma by high-performance liquid chromatography with fluorimetric detection.
A simple and versatile methodology using high-performance liquid chromatography (HPLC) with fluorimetric detection was developed to simultaneously determine d-serine along with other metabolically related neuroactive amino acids in the glutamatergic system: L-serine, L-glutamate, L-glutamine, and glycine. On-column sensitivity was in the lower picomole range. Of two chiral thiol reagents investigated, amino acid derivatives of o-phthaldialdehyde (OPA) in combination with N-isobutyryl-L-cysteine were found to have consistently higher responses than their corresponding N-tert-butyloxycarbonyl-L-cysteine derivatives. This methodology was applied to the quantitative detection of amino acids in human plasma and lays the foundation for further investigations of the role of neuroactive amino acids in the pathophysiology and treatment of neurological and psychiatric disorders.


Evaluating the Impact of Alcohol Policy on Suicide Mortality: A Sex-Specific Time-Series Analysis for Lithuania.
It is reasonable to believe that the alcohol policy environment can impact the suicide mortality rates in a given country, considering the well-known link between alcohol use and death by suicide. The current literature, albeit limited, suggests that an increase in alcohol taxation may result in a decrease in deaths by suicide and that the effect is sex-specific. Therefore, the objective of the current study was to test the impact of three alcohol control policy enactments (in 2008, 2017 and 2018) on suicide mortality among adults 25-74 years of age in Lithuania, by sex. To estimate the unique impact of three alcohol control policies, we conducted interrupted time-series analyses by employing a generalized additive mixed model on monthly sex-specific age-standardized suicide mortality rates from January 2001 to December 2018. Analyses showed a significant impact of the 2017 (p = 0.016) alcohol control policy on suicide mortality for men only. Specifically, we estimated that in the year following the 2017 policy enactment, approximately 57 (95% CI: 9-107) deaths by suicide were prevented among men, 25-74 years of age. The three policy enactments tested were not found to significantly impact the suicide mortality rate among women. Alcohol control policies involving pricing, which result in a notable decrease in alcohol affordability, could be a cost-effective indirect suicide prevention mechanism in not only countries of the former Soviet Union, but in other high-income countries with a comparable health care system to that in Lithuania. HIGHLIGHTSIncreasing excise tax on alcohol was found to have a sex-specific impact on suicide mortalityThe 2017 alcohol policy prevented 57 deaths by suicide among men, 25-74 years of age, in the following yearAlcohol pricing policies may be a cost-effective indirect suicide prevention mechanism.


Calcineurin is required in urinary tract mesenchyme for the development of the pyeloureteral peristaltic machinery.
Congenital obstructive nephropathy is the principal cause of renal failure in infants and children. The underlying molecular and cellular mechanisms of this disease, however, remain largely undetermined. We generated a mouse model of congenital obstructive nephropathy that resembles ureteropelvic junction obstruction in humans. In these mice, calcineurin function is removed by the selective deletion of Cnb1 in the mesenchyme of the developing urinary tract using the Cre/lox system. This deletion results in reduced proliferation in the smooth muscle cells and other mesenchymal cells in the developing urinary tract. Compromised cell proliferation causes abnormal development of the renal pelvis and ureter, leading to defective pyeloureteral peristalsis, progressive renal obstruction, and, eventually, fatal renal failure. Our study demonstrates that calcineurin is an essential signaling molecule in urinary tract development and is required for normal proliferation of the urinary tract mesenchymal cells in a cell-autonomous manner. These studies also emphasize the importance of functional obstruction, resulting from developmental abnormality, in causing congenital obstructive nephropathy.


A retrospective study of the management of pediatric kerion in Trichophyton tonsurans infection.
Kerion celsi is the inflammatory extreme of tinea capitis, representing a delayed hypersensitivity reaction to the causative dermatophyte. Some authors have advocated the use of oral corticosteroids in patients with kerion formation to inhibit the host inflammatory response and minimize the risk of scarring. This retrospective study analyzed the management and outcome of all children younger than 10 years old presenting to our pediatric dermatology service with tinea capitis resulting in kerion formation between 2003 and 2009. We propose that kerion treatment be directed toward the underlying dermatophyte. Oral and intralesional corticosteroids are an unnecessary adjunct to oral antifungal therapy for children with tinea capitis presenting with kerion in urban areas.


Frequency of and factors associated with severe maternal morbidity.
To estimate the frequency of severe maternal morbidity, assess its underlying etiologies, and develop a scoring system to predict its occurrence.Supplemental Digital Content is Available in the Text. This was a secondary analysis of a Eunice Kennedy Shriver National Institute of Child Health and Human Development Maternal-Fetal Medicine Units Network cohort of 115,502 women and their neonates born in 25 hospitals across the United States over a 3-year period. Women were classified as having severe maternal morbidity according to a scoring system that takes into account the occurrence of red blood cell transfusion (more than three units), intubation, unanticipated surgical intervention, organ failure, and intensive care unit admission. The frequency of severe maternal morbidity was calculated and the underlying etiologies determined. Multivariable analysis identified patient factors present on admission that were independently associated with severe maternal morbidity; these were used to develop a prediction model for severe maternal morbidity. Among 115,502 women who delivered during the study period, 332 (2.9/1,000 births, 95% confidence interval 2.6-3.2) experienced severe maternal morbidity. Postpartum hemorrhage was responsible for approximately half of severe maternal morbidity. Multiple patient factors were found to be independently associated with severe maternal morbidity and were used to develop a predictive model with an area under the receiver operating characteristic curve of 0.80. Severe maternal morbidity occurs in approximately 2.9 per 1,000 births, is most commonly the result of postpartum hemorrhage, and occurs more commonly in association with several identifiable patient characteristics. : II.


Two regions of the human platelet F11-receptor (F11R) are critical for platelet aggregation, potentiation and adhesion.
The F11 receptor (F11R) was first identified on the surface of human platelets as a target for a stimulatory monoclonal antibody (M.Ab.F11) that induces secretion, followed by exposure of fibrinogen receptors and aggregation. Cloning of the gene of F11R has revealed that this protein is a cell adhesion molecule (CAM), a member of the Ig superfamily and an ortholog of the murine protein called junctional adhesion molecule (JAM). The present study has identified two domains through which M.Ab.F11 triggers a platelet response culminating with aggregation. M.Ab.F11-mediated platelet adhesion, and the potentiation of collagen and ADP-induced platelet aggregation by M.Ab.F11, were found to involve the same two domains. A F11R recombinant protein (sF11R) completely inhibited platelet aggregation, adhesion and potentiation induced by M.Ab.F11, indicative that the active conformation of the external domain of F11R is present in the soluble, secreted recombinant protein. Furthermore, a specific peptide containing the sequence of the N-terminal amino acids S-1 to C-23 of F11R, and a peptide with the sequence of K-70 to C-82 in the 1st immunoglobulin-like (Ig) fold of F11R, both inhibited M.Ab.F11-induced aggregation, adhesion and potentiation of the aggregation of human platelets. Modeling of the 3D structure of the extracellular domain of the human platelet F11R suggests that these two regions form an active site within the conformation of this CAM. The sequence of these functional domains of F11R (in the N-terminus and 1st Ig-fold) provide the basis for new drug development in the treatment of certain types of thrombocytopenia and inflammatory thrombosis.


Ross River Virus infection--an acute polyarthritis caused by arbovirus.
Epidemic polyarthritis caused by Ross River Virus (RRV) infection has occurred frequently in Australia, and since 1979 has spread into the South Pacific. An outbreak in the Cook Islands in 1980 was investigated and the virus isolated in about one-half of the subjects examined in the acute phases of the illness. Many of the affected adults (but not children) had acute self-limiting polyarthritis which occurred as soon as 3-4 days following infection. RRV infection provides a model for investigation of outbreaks of acute polyarthritis in man and for understanding the pathogenesis of acute viral-induced arthritis.


A novel transcription factor, ERD15 (Early Responsive to Dehydration 15), connects endoplasmic reticulum stress with an osmotic stress-induced cell death signal.
As in all other eukaryotic organisms, endoplasmic reticulum (ER) stress triggers the evolutionarily conserved unfolded protein response in soybean, but it also communicates with other adaptive signaling responses, such as osmotic stress-induced and ER stress-induced programmed cell death. These two signaling pathways converge at the level of gene transcription to activate an integrated cascade that is mediated by N-rich proteins (NRPs). Here, we describe a novel transcription factor, GmERD15 (Glycine max Early Responsive to Dehydration 15), which is induced by ER stress and osmotic stress to activate the expression of NRP genes. GmERD15 was isolated because of its capacity to stably associate with the NRP-B promoter in yeast. It specifically binds to a 187-bp fragment of the NRP-B promoter in vitro and activates the transcription of a reporter gene in yeast. Furthermore, GmERD15 was found in both the cytoplasm and the nucleus, and a ChIP assay revealed that it binds to the NRP-B promoter in vivo. Expression of GmERD15 in soybean protoplasts activated the NRP-B promoter and induced expression of the NRP-B gene. Collectively, these results support the interpretation that GmERD15 functions as an upstream component of stress-induced NRP-B-mediated signaling to connect stress in the ER to an osmotic stress-induced cell death signal.


Effects of Early Acoustic Hearing on Speech Perception and Language for Pediatric Cochlear Implant Recipients.
Purpose The overall goal of the current study was to identify an optimal level and duration of acoustic experience that facilitates language development for pediatric cochlear implant (CI) recipients-specifically, to determine whether there is an optimal duration of hearing aid (HA) use and unaided threshold levels that should be considered before proceeding to bilateral CIs. Method A total of 117 pediatric CI recipients (ages 5-9 years) were given speech perception and standardized tests of receptive vocabulary and language. The speech perception battery included tests of segmental perception (e.g., word recognition in quiet and noise, and vowels and consonants in quiet) and of suprasegmental perception (e.g., talker and stress discrimination, and emotion identification). Hierarchical regression analyses were used to determine the effects of speech perception on language scores, and the effects of residual hearing level (unaided pure-tone average [PTA]) and duration of HA use on speech perception. Results A continuum of residual hearing levels and the length of HA use were represented by calculating the unaided PTA of the ear with the longest duration of HA use for each child. All children wore 2 devices: Some wore bimodal devices, while others received their 2nd CI either simultaneously or sequentially, representing a wide range of HA use (0.03-9.05 years). Regression analyses indicate that suprasegmental perception contributes unique variance to receptive language scores and that both segmental and suprasegmental skills each contribute independently to receptive vocabulary scores. Also, analyses revealed an optimal duration of HA use for each of 3 ranges of hearing loss severity (with mean PTAs of 73, 92, and 111 dB HL) that maximizes suprasegmental perception. Conclusions For children with the most profound losses, early bilateral CIs provide the greatest opportunity for developing good spoken language skills. For those with moderate-to-severe losses, however, a prescribed period of bimodal use may be more advantageous for developing good spoken language skills.


Approach to Patients With High Anion Gap Metabolic Acidosis: Core Curriculum 2021.
The anion gap (AG) is a mathematical construct that compares the blood sodium concentration with the sum of the chloride and bicarbonate concentrations. It is a helpful calculation that divides the metabolic acidoses into 2 categories: high AG metabolic acidosis (HAGMA) and hyperchloremic metabolic acidosis-and thereby delimits the potential etiologies of the disorder. When the [AG] is compared with changes in the bicarbonate concentration, other occult acid-base disorders can be identified. Furthermore, finding that the AG is very small or negative can suggest several occult clinical disorders or raise the possibility of electrolyte measurement artifacts. In this installment of AJKD's Core Curriculum in Nephrology, we discuss cases that represent several very common and several rare causes of HAGMA. These case scenarios highlight how the AG can provide vital clues that direct the clinician toward the correct diagnosis. We also show how to calculate and, if necessary, correct the AG for hypoalbuminemia and severe hyperglycemia. Plasma osmolality and osmolal gap calculations are described and when used together with the AG guide appropriate clinical decision making.


Acromioclavicular joint reconstruction using the Nottingham Surgilig: a preliminary report.
Eleven patients with a chronic acromio-clavicular joint disruption underwent acromio-clavicular joint stabilisation using the Nottingham Surgilig and were followed up clinically and radiologically for an average of 24 months post operation. The mean post-operative Constant score was 83.1, the Imatani score was 81.2 and the Walsh score was 14.1. Eighty-two per cent of patients were satisfied with the operation. There was one case of rupture through the central portion of the Surgilig, and following laboratory analysis, the ligament has been modified since. In 4 patients there was evidence of loosening of the screw but only one complained of this being a problem. This short-term outcome analysis of the Nottingham Surgilig is the first such report outside the Nottingham unit where the prosthesis was originally developed. Our results are encouraging and justify further use and evaluation of this relatively new technique.


A qualitative study of the quality of life of children receiving intravenous nutrition at home.
To discover the views of children with severe intestinal failure treated with intravenous nutrition from early life and who remained heavily dependent on treatment throughout childhood. Seven children ages 7 to 17 years (mean 13 years) were interviewed. The diagnoses were enteropathy in 3, extreme short gut in 1, complex (associated mucosal inflammation and dysmotitlity) in 2, and intestinal pseudo-obstruction in 1. They were treated with intravenous nutrition overnight at home that was administered by trained parents using the simplest possible system. The children were individually questioned about their lifestyle and health. Transcripts were analysed using interpretive phenomenological analysis. Children coped well with life with intravenous nutrition (apart from septicaemia in 2 cases), but were troubled when complications of the underlying disease persisted (eg, nocturnal disturbance, stool frequency, abdominal pain). Children were aware that life was restricted (eg, fewer sleepovers with friends, fewer late nights out). There was a high level of family functioning. Older children wished to take care of themselves. The burdens of life with intravenous nutrition appear to be less significant for these children than living with the effects of chronic illness. There was resilience and acceptance in the face of illness-related demands. This study has found that despite the problems they may face, it is possible for children fed intravenously at home to develop a level of resilience, maintain a positive outlook, and cope well with illness-related demands even when they have had virtually lifelong severe intestinal failure. Families can continue to function well.


Rare BRAF mutations in pancreatic neuroendocrine tumors may predict response to RAF and MEK inhibition.
The clinical significance of BRAF alterations in well-differentiated (WD) metastatic pancreatic neuroendocrine tumor (panNET) is unknown, but BRAF-mutated panNET could represent a subset characterized by an identifiable and clinically actionable driver. Following the identification of two patients with WD metastatic panNET whose tumors harbored BRAF mutations, we queried the MSK-IMPACT series of 80 patients with WD metastatic panNET for additional mutations in BRAF, and in other genes involved in RAS/ RTK/ PI3K signaling pathways. BRAF mutations were identified in six samples (7.5%): two tumors harbored V600E mutations, one tumor each expressed K601E, T599K, and T310I mutations, and one tumor expressed both G596D and E451K BRAF. Few additional actionable driver alterations were identified. To determine the ERK activating capability of four BRAF mutations not previously characterized, mutant constructs were tested in model systems. Biochemical characterization of BRAF mutations revealed both high and low activity mutants. Engineered cells expressing BRAF K601E and V600E were used for in vitro drug testing of RAF and MEK inhibitors currently in clinical use. BRAF K601E demonstrated reduced sensitivity to dabrafenib compared to BRAF V600E, but the combination of RAF plus MEK inhibition was effective in cells expressing this mutation. Herein, we describe the clinical course of a patient with BRAF K601E and a patient with BRAF V600E WD metastatic panNET, and the identification of four mutations in BRAF not previously characterized. The combined clinical and biochemical data support a potential role for RAF and MEK inhibitors, or a combination of these, in a selected panNET population.


[Repair of whole-hand destructive injury and hand degloving injury with transplant of pedis compound free flap].
To investigate the surgical method and clinical efficacy of repairing whole-hand destructive injury or hand degloving injury with the transplant of pedis compound free flap. From February 2003 to June 2008, 21 patients with whole-hand destructive injury or hand degloving injury were treated, including 15 males and 6 females aged 18-45 years old (average 25 years old). The injury was caused by punching machine crush in 10 cases, roller crush in 7 cases, and imprinter crush in 4 cases. The time between injury and operation was 1-9 hours. Eleven cases had the skin-degloving injury of the whole hand, while the other 10 cases had the proximal palm injury combined with dorsal or palmar skin and soft tissue defect. After debridement, the size of wound was 9 cm x 7 cm-15 cm x 10 cm in the dorsal aspect and 10 cm x 7 cm-6 cm x 10 cm in the palmar aspect. The defect was repaired by the thumbnail flap of dorsalis pedis flap and the second toenail flap of dorsalis pedis flap in 5 cases, the thumbnail flap of dorsalis pedis flap and the second toe with dorsalis pedis flap in 4 cases, and bilateral second toe with dorsalis pedis flap in 12 cases. The flap area harvested during operation ranged from 6 cm x 5 cm to 16 cm x 11 cm. Three fingers were constructed in 2 cases and two fingers in 19 cases. Distal interphalangeal joint toe amputation was conducted in the thumbnail flap donor site, metatarsophalangeal joint toe amputation was performed in the second toenail flap donor site, and full-thickness skin grafting was conducted in the abdomen. At 7 days after operation, the index finger in 1 case repaired by the second toenail flap suffered from necrosis and received amputation, 1 case suffered from partial necrosis of distal dorsalis pedis flap and recovered after dressing change, and the rest 42 tissue flaps survived. Forty-three out of 44 reconstructed fingers survived. All the wounds healed by first intention. At 2 weeks after operation, 2 cases had partial necrosis of the donor site flap and underwent secondary skin grafting after dressing change, the rest skin grafts survived, and all the wounds healed by first intention. Nineteen cases were followed up for 6-36 months (average 11 months). The flaps of palm and dorsum of hand showed no swelling, the reconstructed fingers had a satisfactory appearance and performed such functions as grabbing, grasping, and nipping. The sensory of the flaps and the reconstructed fingers recovered to S2-S4 grade. The donor site on the dorsum of the foot had no obvious scar contracture, without obvious influence on walking. For the whole-hand destructive injury or hand degloving injury, the method of transplanting pedis compound free flap can repair the defect in the hand and reconstruct the function of the injured hand partially. It is an effective treatment method.


Bismuth classification is associated with the requirement for multiple biliary drainage in preoperative patients with malignant perihilar biliary stricture.
Single preoperative biliary drainage for malignant perihilar biliary stricture occasionally fails to control jaundice and cholangitis. Multiple biliary drainage is required in such cases, but their clinical background is unclear. We determined the clinical characteristics associated with the requirement for multiple biliary drainage. The consecutive 122 patients with malignant perihilar biliary stricture were enrolled in a single-center retrospective study. Preoperative biliary drainage was initially performed on the future remnant hepatic lobe. Additional drainage was performed if jaundice failed to improve or cholangitis developed in undrained hepatic lobes. Detailed clinical characteristics and the number of preoperative biliary drainage procedures required before operation were analyzed. Thirty-one patients (25.4%) initially underwent multiple biliary drainage. However, 69 (56.7%) required multiple biliary drainage by the time of the operation. In the univariate analysis, the initial serum bilirubin level, cholangitis, percutaneous portal vein embolization, history of inserted endoscopic biliary stenting, length of preoperative period, operative procedure, and Bismuth classification were significant factors. In the multivariate analysis using these factors, Bismuth classification was independently associated with the requirement for multiple biliary drainage. The number of patients who required multiple biliary drainage was higher in those with Bismuth-II (91.9%), Bismuth-IIIa (65.7%), and Bismuth-IV (92.9%) than in those with Bismuth-I (22.2%) and Bismuth-IIIb (18.2%). Patients with Bismuth-II, Bismuth-IIIa, and Bismuth-IV are at higher risk for multiple biliary drainage. A strategy based on the Bismuth classification for performing preoperative biliary drainage is important for patients with malignant perihilar biliary stricture.


Frequent Antiviral Treatment Failures in Patients Infected With Hepatitis C Virus Genotype 4, Subtype 4r.
Hepatitis C virus (HCV) genotype 4 is highly heterogeneous. HCV subtype 4r has been suggested to be less responsive to direct-acting antiviral (DAA) drug treatment than other genotype 4 subtypes. Among 537 DAA-treated patients who experienced a virological failure (VF) in France between 2015 and 2018, 121 (22.5%) were infected with genotype 4 and 27 of them (22.3%) with subtype 4r; subtype 4r was thus over-represented as compared to its prevalence in the French general population. Population sequencing of the nonstructural protein (NS) 3, NS5A, and NS5B genes was performed in all subtype 4r patients at treatment failure and in 6 at baseline, whereas full-length HCV genome sequencing was performed in two baseline and three treatment failure samples by means of an original shotgun metagenomics method based on deep sequencing. At treatment failure, all subtype 4r patients harbored two to three dominant NS5A resistance-associated substitutions (RASs), including at least L28A/C/I/M/V and L30R. Among 13 patients exposed to sofosbuvir and an NS5A inhibitor (daclatasvir, ledipasvir, or velpatasvir), 5 (38.5%) also harbored NS5B S282C/T RASs at treatment failure. An additional patient harbored S282C/T RASs at treatment failure by deep sequencing. Prevalence of S282C/T RASs at treatment failure was significantly higher in patients infected with genotype 4r than with other genotypes, including other subtypes of genotype 4. Conclusion: The lower rates of sustained virological response in patients infected with subtype 4r are related to the frequent preexistence at treatment baseline and subsequent selection by DAA treatment of both NS5A and NS5B S282 RASs. Our study suggests that these patients should be identified and receive a triple DAA combination regimen as first-line treatment.


Errors and pitfalls: Briefing and accusation of medical malpractice - the second victim.
In June 2012, the German Medical Association (Bundesärztekammer) published the statistics of medical malpractice for 2011 (published at http://www.bundesaerztekammer.de). Still ENT-specific accusations of medical malpractice are by far the fewest in the field of hospitals and actually even in the outpatient context. Clearly most of the unforeseen incidents still occur in the disciplines of trauma surgery and orthopedics. In total, however, an increasing number of errors in treatment can be noticed on the multidisciplinary level: in 25.5% of the registered cases, an error in treatment was found to be the origin of damage to health justifying a claim for compensation of the patient. In the year before, it was only 24.7%. The reasons may be manifold, but the medical system itself certainly plays a major role in this context: the recent developments related to health policy lead to a continuous economisation of medical care. Rationing and limited remuneration more and more result in the fact that therapeutic decision are not exclusively made for the benefit of the patient but that they are oriented at economic or bureaucratic aspects. Thus, in the long term, practising medicine undergoes a change. According to the §§ 1, 3 of the professional code of conduct for doctors (Musterberufsordnung für Ärzte; MBO-Ä) medical practice as liberal profession is principally incompatible with the pursuit of profit, however, even doctors have to earn money which more and more makes him play the role of a businessman. Lack of personnel and staff savings lead to excessive workloads of physicians, caregivers, and nurses, which also favour errors. The quality and even the confidential relationship between doctor and patient, which is important for the treatment success, are necessarily affected by the cost pressure. The victims in this context are not only the patients but also the physicians find themselves in the continuous conflict between ethical requirements of their profession and the actual requirements of the realities in the healthcare field. But also the technical and scientific progress bear new risks beside the therapeutic successes, further especially bigger hospitals require high efforts regarding organisation favouring errors in cases of deficiencies. Even the increasing juridification of the medicine that is expected to achieve a provisional highlight with the planned law of patients' rights leads to an important focus on the quality of medical care (see also [1]). The explicit legal regulation of patients' rights, which have never been out of question up to now, confirms the impression of patients who have to be protected from their doctors. This development favours a natural mistrust in the quality of the treatment and the desire of legal verification in cases of treatment failures. A totally perfect and error-free treatment, however, will never occur. Already this fact leads to the obligation to do everything possible to reduce the risk to an absolute minimum. The risks that might arise from a relation of treatment are manifold. Not only may the patient undergo risks that arise in particular from lacking or insufficient briefing, complications, or medical malpractice. Also the doctor has to fear legal consequences if he does not stick clearly to the increasing requirements that jurisdiction and legislation impose - not least by the planned law of patients' rights. In the following, the basic principles and particularities will be described that apply for the patients' briefing. Further the different types of medical malpractice will be explained in relation to the resulting procedural consequences. Finally some current problematic fields will be described with regard to other possible liabilities or responsibilities of physicians in hospitals or doctor's offices.


Prospective analysis of clinician accuracy in the diagnosis of benign anorrectal pathology: the value of clinical information.
the prevalence of anorectal disorders in general population is high. The aim of this study was to analyze the influence of clinical symptoms on diagnostic accuracy for benign anorectal pathology among different specialists and evaluate the relationship between diagnostic accuracy and years of professional experience. seven typical cases were selected. In a first interview, participants were shown images and asked to make a diagnosis. Afterwards, images with additional information (clinical symptoms) were used. Two groups (group 1 = general surgeons and group 2 = medical specialists who attended emergency department) completed both phases of the study to analyze the influence of clinical symptoms on the final diagnosis. forty four specialists were interviewed. The percentage of participants making a correct diagnosis in groups 1 and 2, respectively, was as follows: case 1 (perianal abscess): 100 vs. 80.6%, (p = 0.157); case 2 (fissure): 92.3 vs. 51.6% (p = 0.015); case 3 (thrombosed hemorrhoid): 92.3 vs. 74.2% (p = 0.321); case 4 (anal condyloma): 100 vs. 87.1% (p = 0.302); case 5 (rectal prolapse): 100 vs. 83.9% (p = 0.301); case 6 (prolapsed hemorrhoid): 92.3 vs. 29% (p = 0.001), and case 7 (fistula): 100 vs. 67.7% (p = 0.021). There were significant differences in the number of correctly diagnosed cases between groups (p < 0.001). Information about clinical symptoms significantly increased overall and specific accuracy. There was no correlation between experience and accuracy. clinical symptoms are important for diagnostic accuracy in anorectal pathology. Training in anorectal pathology in medical specialists is warranted.


Statistical analysis of dislocations and dislocation boundaries from EBSD data.
Electron BackScatter Diffraction (EBSD) is often used for semi-quantitative analysis of dislocations in metals. In general, disorientation is used to assess Geometrically Necessary Dislocations (GNDs) densities. In the present paper, we demonstrate that the use of disorientation can lead to inaccurate results. For example, using the disorientation leads to different GND density in recrystallized grains which cannot be physically justified. The use of disorientation gradients allows accounting for measurement noise and leads to more accurate results. Misorientation gradient is then used to analyze dislocations boundaries following the same principle applied on TEM data before. In previous papers, dislocations boundaries were defined as Geometrically Necessary Boundaries (GNBs) and Incidental Dislocation Boundaries (IDBs). It has been demonstrated in the past, through transmission electron microscopy data, that the probability density distribution of the disorientation of IDBs and GNBs can be described with a linear combination of two Rayleigh functions. Such function can also describe the probability density of disorientation gradient obtained through EBSD data as reported in this paper. This opens the route for determining IDBs and GNBs probability density distribution functions separately from EBSD data, with an increased statistical relevance as compared to TEM data. The method is applied on deformed Tantalum where grains exhibit dislocation boundaries, as observed using electron channeling contrast imaging.


Evaluating efficacy of pharmaceutical interventions in atherosclerosis: role of magnetic resonance imaging and positron emission tomography.
The rate of acute complications of atherosclerosis (acute myocardial infarction, ischemic stroke) has continuously decreased over the last 20 years in Western countries. This is largely explained by improvements in the reduction and treatment of cardiovascular risk factors and by the increasing number of patients who benefit from preventive treatments such as antiplatelet, lipid-lowering, or antihypertensive drugs. This means also that, when testing new drugs aimed at either halting or even reversing the progression of atherosclerotic plaques, a large number of patients will need to be included in clinical trials to demonstrate an improvement in patient outcome with the drugs. Pharmaceutical companies are therefore looking for early surrogate markers that could be evaluated in a small number of patients to predict the beneficial effects of new drugs on atherosclerotic plaques before moving to costly clinical trials with a large number of patients. In this review, we will discuss the place of atherosclerotic plaque imaging with magnetic resonance imaging and positron emission tomography for the evaluation of new antiatherosclerotic drugs.


Taming the blackbody with infrared metamaterials as selective thermal emitters.
In this Letter we demonstrate, for the first time, selective thermal emitters based on metamaterial perfect absorbers. We experimentally realize a narrow band midinfrared (MIR) thermal emitter. Multiple metamaterial sublattices further permit construction of a dual-band MIR emitter. By performing both emissivity and absorptivity measurements, we find that emissivity and absorptivity agree very well as predicted by Kirchhoff's law of thermal radiation. Our results directly demonstrate the great flexibility of metamaterials for tailoring blackbody emission.


Calcium supplementation with calcium-rich mineral waters: a systematic review and meta-analysis of its bioavailability.
The relevance of calcium (Ca2+), an essential bone mineral, to the prevention and treatment of osteoporosis is well established. However, a good deal of evidence casts doubt on the validity of current RDAs (recommended daily allowance), i.e., 800-1000 mg/day. New guidelines consistently advocate higher daily intakes (up to 1500 mg/day), a goal that may be difficult to achieve for many patients. Environmental as well as individual behavioral factors may limit the consumption of dairy products, whereas calcium supplements require a high level of compliance and cause additional costs. Calcium-rich mineral waters may offer a promising alternative. A systematic literature search was performed (Medline, years 1966-1998) to identify experimental studies on the bioavailability of calcium-rich mineral waters. First, all publications on calcium absorption from mineral waters were identified, and, in a second step, studies comparing calcium absorption from mineral waters with that from dairy products. Four studies fulfilled all inclusion criteria. A meta-analysis based on published p values indicated calcium absorption from mineral waters was significantly higher (p = 0.03) than that from dairy products. Although only few studies with a relatively small number of subjects are available to date, the bioavailability of calcium from calcium-rich mineral waters thus seems to be at least comparable to, and possibly better than, that from dairy products. These results are in keeping with the assumption that calcium-rich mineral water is a useful calcium source to achieve new, higher recommended daily allowances of calcium.


Mitochondria-targeted antioxidant peptides.
Overproduction of reactive oxygen species (ROS) under pathophysiologic conditions is part of the disease process. These ROS are released from different sources, and in particular from mitochondria. Although the molecular mechanisms responsible for mitochondria-mediated disease processes are unclear, oxidative stress seems to play an important role. ROS are essential to cell function, but adequate levels of antioxidant defenses are required in order to avoid the harmful effects that excessive ROS production can produce. Mitochondrial oxidative stress damage and dysfunction contribute to a number of cell pathologies that manifest themselves through a range of conditions. The antioxidants available until now have not proved to be particularly effective against many of these disorders. It is possible that these antioxidants do not reach the sites of free radical generation, especially when mitochondria are the primary source of ROS. Recent developments in mitochondria-targeted antioxidants have moved closer to providing protection against mitochondrial oxidative damage. The SS (Szeto-Schiller) peptide antioxidants represent a novel approach that employs the targeted delivery of antioxidants to the inner mitochondrial membrane. These SS peptides scavenge hydrogen peroxide and peroxynitrite and inhibit lipid peroxidation. By reducing mitochondrial ROS, they inhibit mitochondrial permeability transition and cytochrome c release, thus preventing oxidant-induced cell death. Preclinical studies support the use of these peptides for ischemia-reperfusion injury and neurodegenerative disorders. Although peptides have often been considered to be poor drug candidates, the few that have been studied are promising agents for the treatment of diseases.


Activated human platelets express beta2 integrin.
The expression of molecules of the beta2 integrin family (CD11a, CD11b , CD11c, and CD18) was explored on 2 human megakaryocytic cell lines and on platelets from different donors by immunofluorescence and flow cytometry using a large panel of mAb. CD11a, CD11b, CD11c and CD18 were detected on the megakaryocytic cell lines DAMI and HEL. A low and variable expression of CD11a, CD11b and CD18 determinants was also detected on resting platelets; this expression was markedly increased when platelets were activated by thrombin. Expression of CD18 was closely correlated to that of CD11a or CD11b when comparing the fluorescence intensity observed in different experiments. In presence of Ca++, platelets did bind to a RAJI cell line which exhibits a high expression level of CD54. This binding was increased when platelets were activated by thrombin and was decreased by an anti CD11a, CD18 and anti CD54 mAb. This study indicates that human platelets express molecules of the beta2 integrin family, when activated, which allows them to bind to CD54 bearing cells.


Regulation of breast cancer cell behaviours by the physical microenvironment constructed via projection microstereolithography.
A considerable number of studies have examined how intrinsic factors regulate breast cancer cell behaviours; however, physical microenvironmental cues may also modulate cellular morphology, proliferation, and migration and mechanical properties. In the present study, the surrounding microenvironment of breast cancer cells was constructed using projection microstereolithography, enabling the investigation of the external environment's effects on breast cancer cell behaviours. A poly(ethylene) glycol diacrylate (PEGDA) solution was polymerized by programmable ultraviolet exposure to create arbitrary shapes with high biocompatibility, efficiency, flexibility and repeatability, and the resistance to cell attachment enabled the PEGDA coated film to hinder cell adhesion, allowing cells to grow in specific patterns. Furthermore, breast cancer cell morphology and mechanical properties were modified by altering the microenvironment. Proliferation was higher in breast cancer as compared to normal cells, consistent with the primary characteristic of malignant tumors. Moreover, breast cancer cells migrated more rapidly when grown in a narrow channel as compared to a wider channel. These findings enhance our understanding of the role of the microenvironment in breast cancer cell behaviours and can provide a basis for developing effective anticancer therapies.


Effects of family-centered care on bonding: A systematic review.
Most critically ill neonates require constant monitoring, continuous care, and supervision. However, distance created by admission and prolonged stay in a neonatal intensive care unit (NICU) may contribute to a delay in parent-infant bonding. This review aimed to determine how family-centered care (FCC) in the NICU affects parental bonding with critically ill infants. The Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines were used to perform a systematic search of the literature within the following four electronic databases: Cumulative Index of Nursing and Allied Health Literature (CINAHL), MEDLINE, Cochrane Library, and Web of Science. The search was conducted through July/August 2020. Research quality was evaluated using the Johns Hopkins Nursing Evidence-Based Practice Grading Scale. Out of 816 articles identified through literature search, 16 of the studies met our inclusion criteria. The majority of the studies (n = 14) found FCC interventions resulted in a significant increase in parental bonding. Results showed evidence practicing FCC in the NICU setting supports early parent-infant bonding. Nurses should consider implementing evidence-based FCC strategies into practice, such as allowing parents unrestricted access to their infants. More rigorous research with larger samples is recommended. More studies are also needed focusing on father-infant dyads and mother-father-infant triads.


[Effects of acupuncture on the number and degranulation ratio of mast cells and expression of tryptase in synovium of rats with adjuvant arthritis].
To observe the effects of acupuncture on synovial pathology, synovial mast cell degranulation and tryptase expression and to investigate the relationship between the functions of mast cells and effects of acupuncture on early adjuvant arthritis in rats. Forty-six male Wistar rats were randomly divided into normal control group (n=16), untreated group (n=15) and acupuncture group (n=15). Adjuvant arthritis was induced by injection of 0.1 mL Freund's complete adjuvant in right hind limb footpad. Normal control group and untreated group received no acupuncture treatment, while rats in the acupuncture group were treated with sterilized disposable stainless steel needles inserted perpendicularly as deep as 2 to 3 mm at Xuanzhong (GB39), 6 mm at Shenshu (BL23) and 7 mm at Zusanli (ST36) for eight times (15 min each time) every two days. Setting the modeling day as the 0 day of the experiment, the body weight and paw volume of the rats were measured every three days from the 0 day. In the end, synovial tissues of the right hind ankles were sampled and made into paraffin sections. Then they were firstly stained with hematoxylin-eosin for observing synovial pathology to evaluate the effects of acupuncture on adjuvant arthritis, then stained with toluidine blue for observing the number and degranulation ratio of synovial mast cells and finally detected by immunohistochemical staining method to investigate the expression of tryptase in synovium. Compared with the untreated group, the body weight of rats in the acupuncture group was increased significantly (P<0.05), while the paw volume decreased obviously (P<0.01). Hematoxylin-eosin staining showed that acupuncture significantly inhibited inflammatory cell infiltration, synovial cell hyperplasia, and synovial fibroplasia in synovium of rats with adjuvant arthritis as compared with the untreated group (P<0.05). Toluidine blue staining showed that acupuncture could significantly diminish the numbers of total and degranulated mast cells in rats with adjuvant arthritis (P<0.01), which were significantly higher in the untreated group than in the normal control group (P<0.01). Showing by immunohistochemical staining, the expression of tryptase in synovium in the acupuncture group was decreased as compared with the untreated group (P<0.01). Analyzed by Spearman's bivariate correlation, the number of mast cells and degranulation ratio of mast cells were positively correlated with the pathological scores (r=0.837, P<0.01; r=0.634, P<0.01). Acupuncture can improve pathological condition of inflammatory synovium in rats with early adjuvant arthritis by inhibiting the function of synovial mast cells, which may play an important underlying role in the immunoregulation of acupuncture on adjuvant arthritis.


Dynamic associations between opioid use and anhedonia: A longitudinal study in opioid dependence.
Anhedonia is a commonly reported symptom among substance-dependent populations that appears to diminish with sustained abstinence. However, previous research has not determined whether anhedonia is dynamically linked to changing patterns of drug use, nor whether it predicts subsequent drug use. We aimed to test whether changes in illicit opioid use would predict changes in anhedonia, and whether increases in anhedonia would predict further opioid use. We conducted a longitudinal, observational study, with a convenience sample of 121 participants with current or past-year Diagnostic and Statistical Manual for Mental Disorders, Fourth Edition Text Revision (DSM-IV-TR) opioid dependence recruited from substance use treatment and related services and from pharmacies administering opioid substitution pharmacotherapy. Anhedonia was assessed with the Temporal Experience of Pleasure Scale and frequency of illicit opioid use was assessed using timeline follow-back interviews. There was a significant within-subject effect (β=-0.015; 95% CI -0.02 to -0.01; p=0.001), indicating that participants' Temporal Experience of Pleasure Scale scores typically declined (i.e. anhedonia increased) following a month with above-average opioid use and Temporal Experience of Pleasure Scale scores rose (i.e. anhedonia reduced) following a month with below-average opioid use. However, Temporal Experience of Pleasure Scale scores did not significantly predict opioid use in the subsequent month (β=-0.04, 95% CI -0.20 to 0.12; p=0.651). Changes in illicit opioid use predict self-reported anhedonia, suggesting a possible causal relationship whereby anhedonia is likely to worsen with frequent drug use and diminish with prolonged abstinence. However, anhedonia does not appear to drive further drug use.


Chromosomal transmission bias in laboratory hybrids between wild strains of the two European subspecies of house mice.
Laboratory crosses between wild strains of the two European house mouse subspecies Mus musculus domesticus (2n = 34) and M. m. musculus (2n = 40) were performed to analyze the selective processes involved in the non-introgression of centromeric regions of Robertsonian (Rb) fusions in the Danish hybrid zone. The chromosomal analysis of 226 backcross progeny from 22 reciprocal crosses showed that the segregation of the three Rb fusions present did not significantly differ from Mendelian expectations. However, a significant negative correlation was found between Rb transmission rates and the average litter sizes of the F1 pairs. Among the different models of selection discussed, the most likely one supported the existence of two opposing selective factors resulting in an overall compensation of chromosomal types in the backcross progeny. A two-phase selective process involving embryo competition was postulated with non-Rb carriers being favored during pre-implantation but disadvantaged after implantation. Such balanced selective pressures acting on musculus non-Rb centromeres are compatible with the steep slope and off-centered position of the chromosomal cline observed in the Danish hybrid zone. These results suggested that these selective factors may be more related to centromere origin (musculus or domesticus) than to centromere structure (Rb or non-Rb).


European cardiovascular mortality over the last three decades: evaluation of time trends, forecasts for 2016.
The circulatory diseases, in particular ischemic heart diseases and stroke, represent the main causes of death worldwide both in high income and in middle and low income countries. Our aim is to provide a comprehensive report to depict the circulatory disease mortality in Europe over the last 30 years and to address the sources of heterogeneity among different countries. Our study was performed using the WHO statistical information system - mortality database - and was restricted to the 28 countries belonging to the European Union (EU-28). We evaluated gender and age time series of all circulatory disease mortality, ischemic heart diseases, cerebrovascular diseases, pulmonary and other circulatory diseases and than we performed forecast for 2016. Mortality heterogeneity was evaluated by countries using the Cochrane Q statistic and the I-squared index. Between 1985 and 2011 SDR for deaths attributable to all circulatory system diseases decreased from 440.9 to 212.0 x 100,000 in EU-28 and a clear uniform reduction was observed. Heterogeneity among countries was found to be consistent, therefore different analysis were carried out considering geographical area. We forecast a reduction in European cardiovascular mortality. Heterogeneity among countries could only in part be explained by both geographical and health expenditure factors.


Interacting effects of toxicants and organic matter on the midge Chironomus riparius in polluted river water.
Toxicants and organic matter in river water have contrasting impacts on macrofauna. Through manipulations of both factors, their interactive effects on organisms were evaluated. This way, an attempt was made to clarify the presence or absence of pollution-"tolerant" and -"sensitive" species in rivers affected by mixed sources of pollution. Under controlled conditions, larval growth of the "tolerant" midge Chironomus riparius was measured in different types of river water containing varying levels of particles (obtained by selective filtration) and toxicants (either complex mixtures or metals). Exposure of first-instar larvae to water from the polluted rivers Meuse and Dommel revealed that growth was less inhibited by toxicant levels in river water than expected based on laboratory toxicity tests. Factors present in polluted river water stimulated growth of midges to such an extent that inhibiting effects of high toxicant concentrations were neutralized, and at low toxicant levels, were overcompensated for. It was found that particulate matter has great potential to reduce inhibiting effects of toxicants on C. riparius, not (only) by reducing the bioavailability of toxicants, but by serving as a supplementary, superior food source. The success of the "pollution-tolerant" midge was not explained by tolerance of this species to toxicants, but by its ability to take advantage of coinciding organic enrichment. It is hypothesized that the extent to which beneficial effects of organic compounds on organisms occur is species specific.


A high rate of novel CYP11B1 mutations in Saudi Arabia.
Despite ethnic variation, 11 β-hydroxylase deficiency (11β-OHD) has generally been considered the second most common subtype of congenital adrenal hyperplasia (CAH). We report a high rate of novel mutations in this gene (CYP11B1) in patients from Saudi Arabia. We studied 16 patients with 11β-OHD from 8 unrelated families. DNA was isolated from peripheral blood. The 9 exons and exon-intron boundaries of CYP11B1 were PCR-amplified and directly sequenced. The novel mutations were functionally characterized using subcloning, in vitro mutagenesis, cell transfection and 11-deoxycortisol: cortisol conversion assays. Six mutations were found in these 8 unrelated families. Three of these mutations are completely novel and two have just been recently described as novel mutations from the same population. These include a single nucleotide insertion mutation in codon 18 (c.53_54insT) leading to frameshift and truncation in 4 siblings, a novel mutation (c.1343G>C, p.R448P) in 3 unrelated families, a novel mutation (c.1394A>T, p.H465L) in 2 siblings, a novel mutation (c.617G>T, p.G206V) in 1 patient, and a recently described non-sense novel mutation (c.780G>A, p.W260X) in another patient. Out of the 6 mutations described in this report, only one mutation (p.Q356X) was reported previously. In vitro functional testing of the 3 missense and nonsense novel mutations revealed complete loss of the 11 hydroxylase activity. We conclude that 11 β-OHD in Saudi Arabia has a unique genotype with a high rate of novel mutations. The novel p. R448P mutation is the most common mutation in this highly inbred population.


The PRIMARY Score: Diagnostic Performance and Added Value Compared With MRI in Detecting Clinically Significant Prostate Cancer.
Multiparametric MRI is the current standard for detecting clinically significant prostate cancer (csPCa). However, men with negative or equivocal MRI often undergo unnecessary biopsies due to concerns about false-negative results. The recently proposed 68 Ga-PSMA PET/CT-based PRIMARY score exhibited good diagnostic performance for csPCa. This study aimed to externally validate the performance of the PRIMARY score and evaluate its added diagnostic value to MRI triage in detecting csPCa. This retrospective cohort study included 431 men who underwent both 68 Ga-PSMA PET/CT and MRI before biopsy. Performance was assessed using the area under the receiver operating characteristic curve and the decision curve analysis. The PRIMARY score + MRI was considered positive for either PRIMARY score 3-5 or Prostate Imaging Reporting and Data System (PI-RADS) 4/5. The prevalence of csPCa was 51.7% (223/431). The area under the receiver operating characteristic curve of the 5-level PRIMARY score for csPCa was significantly higher than that of MRI (0.873 vs 0.786, P < 0.001). For the entire group, sensitivity, specificity, positive predictive value, and negative predictive value of the PRIMARY score were 90.6%, 61.1%, 71.4%, and 85.8%, respectively, which outperformed 87.9%, 49.0%, 64.9%, and 79.1% of PI-RADS on MRI. The PRIAMRY score + MRI improved sensitivity (96.0% vs 87.9%, P < 0.001) and negative predictive value (91.5% vs 79.1%, P < 0.001) without compromising specificity and positive predictive value compared with MRI alone. This combined approach avoided 24.6% (106/431) of unnecessary biopsies, while missing 4.0% (9/223) of csPCa cases. The addition of the PRIMARY score in men with PI-RADS 1-3 showed a net benefit, but not in men with PI-RADS 4/5. The PRIMARY score was superior to MRI in detecting csPCa, and its added diagnostic value was in men with negative or equivocal MRI results. The PRIMARY score + MRI improved negative predictive value and sensitivity for csPCa compared with MRI alone. Further prospective trials will validate whether men with clinical suspicion of csPCa but negative PRIMARY score + MRI can safely avoid unnecessary biopsies.


Pullulan encapsulation of labile biomolecules to give stable bioassay tablets.
A simple and inexpensive method is reported for the long-term stabilization of enzymes and other unstable reagents in premeasured quantities in water-soluble tablets (cast, not compressed) made with pullulan, a nonionic polysaccharide that forms an oxygen impermeable solid upon drying. The pullulan tablets dissolve in aqueous solutions in seconds, thereby facilitating the easy execution of bioassays at remote sites with no need for special reagent handling and liquid pipetting. This approach is modular in nature, thus allowing the creation of individual tablets for enzymes and their substrates. Proof-of-principle demonstrations include a Taq polymerase tablet for DNA amplification through PCR and a pesticide assay kit consisting of separate tablets for acetylcholinesterase and its chromogenic substrate, indoxyl acetate, both of which are highly unstable. The encapsulated reagents remain stable at room temperature for months, thus enabling the room-temperature shipping and storage of bioassay components.


Assessment of the local SAR distortion by major anatomical structures in a cylindrical neck phantom.
The objective of this work is to gain insight in the distortions on the local SAR distribution by various major anatomical structures in the neck. High resolution 3D FDTD calculations based on a variable grid are made for a semi-3D generic phantom based on average dimensions obtained from CT-derived human data and in which simplified structures representing trachea, cartilage, spine and spinal cord are inserted. In addition, phantoms with dimensions equal to maximum and minimum values within the CT-derived data are also studied. In all cases, the phantoms are exposed to a circular coherent array of eight dipoles within a water bolus and driven at 433 MHz. Comparisons of the SAR distributions due to individual structures or a combination of structures are made relative to a cylindrical phantom with muscle properties. The calculations predict a centrally located region of high SAR within all neck phantoms. This focal region, expressed as contours at either 50% or 75% of the peak SAR, changes from a circular cross-section in the case of the muscle phantom to a doughnut shaped region when the anatomical structures are present. The presence of the spine causes the greatest change in the SAR distribution, followed closely by the trachea. Global changes in the mean SAR relative to the uniform phantom are <11%, whilst local changes are as high as 2.7-fold. There is little difference in the focal dimensions between the average and smallest phantoms, but a decrease in the focal region is seen in the case of the largest phantom. This study presents a first step towards understanding of the complex influences of the various parameters on the SAR pattern which will facilitate the design of a site-specific head and neck hyperthermia applicator.


[Experimental Investigation on the Electron Temperature of Laser-Induced Mg Plasmas].
A series of plasma characteristic spectral lines of Mg alloy were obtained under nanosecond laser shock produced by a pulsed Nd : YAG laser (1 064 nm, maximum energy 500 mJ), which was taken under standard atmospheric pressure and at room temperature. Results indicated that the evolutionary rates of spectral lines were different, and the laser energy was enough to ionization Mg alloy under this experimental condition by the spectral lines of MgⅠ, MgⅡ. The electron temperature of Mg plasma were calculated by the measured relative Emission-line intensity(MgⅠ383.2 nm, MgⅠ470.3 nm, MgⅠ518.4 nm). The experimental results showed that the secondary excitation Mg atoms could be got under this experimental condition. The electron temperature of Mg plasma decreased with the laser energy reduced while the laser energy was in the range of 200~500 mJ. When the laser energy was in the range of 350~500 mJ, the electron temperature changed rapidly. When the laser energy was in the range of 200~350 mJ, the electron temperature changed slowly and tended towards stability. It is found that when the laser energy was 300 mJ, the plasma temperature raised suddenly, which could not accord with the trend because of plasma shielding. When the laser energy was 300 mJ, the relative intensity of spectral lines was reduced which was lower than that of 350 and 250 mJ. And it is against the variation trend of the relative intensity of spectral lines increases with the increase of laser energy, which prove plasma shielding phenomenon had occurred and plasma with high power laser separate the coupling between laser and material. The plasma temperature was increased significantly, which is not consistent with the trend .When the plasma shielding phenomenon happened, laser energy was absorbed by the plasma, resulting in the rise of plasma temperature.


Artificial Intelligence-Based Application to Explore Inhibitors of Neurodegenerative Diseases.
Neuroinflammation is a common factor in neurodegenerative diseases, and it has been demonstrated that galectin-3 activates microglia and astrocytes, leading to inflammation. This means that inhibition of galectin-3 may become a new strategy for the treatment of neurodegenerative diseases. Based on this motivation, the objective of this study is to explore an integrated new approach for finding lead compounds that inhibit galectin-3, by combining universal artificial intelligence algorithms with traditional drug screening methods. Based on molecular docking method, potential compounds with high binding affinity were screened out from Chinese medicine database. Manifold artificial intelligence algorithms were performed to validate the docking results and further screen compounds. Among all involved predictive methods, the deep learning-based algorithm made 500 modeling attempts, and the square correlation coefficient of the best trained model on the test sets was 0.9. The XGBoost model reached a square correlation coefficient of 0.97 and a mean square error of only 0.01. We switched to the ZINC database and performed the same experiment, the results showed that the compounds in the former database showed stronger affinity. Finally, we further verified through molecular dynamics simulation that the complex composed of the candidate ligand and the target protein showed stable binding within 100 ns of simulation time. In summary, combined with the application based on artificial intelligence algorithms, we unearthed the active ingredients 1,2-Dimethylbenzene and Typhic acid contained in Crataegus pinnatifida and Typha angustata might be the effective inhibitors of neurodegenerative diseases. The high prediction accuracy of the models shows that it has practical application value on small sample data sets such as drug screening.


Cardiac Failure in a Trisomy 9 Patient Undergoing Anesthesia: A Case Report.
A 27-year-old female with Trisomy 9 mosaicism presented to Children's Hospital Colorado for outpatient dental surgery under general anesthesia. The patient's past medical history was also significant for premature birth, gastroesophageal reflux, scoliosis and kyphosis, obesity, and developmental delay. Per her mother's report, the patient had no cardiac issues. She had undergone multiple previous general anesthetics, some of which documented respiratory complications such as laryngospasm, bronchospasm, and possible aspiration. During this anesthetic, the patient became hypotensive on induction, with sluggish response to intravenous fluids, glycopyrrolate, and ephedrine. Her electrocardiogram demonstrated what appeared to be left bundle branch block at baseline, with possible ST segment changes after induction. Due to her abnormal reaction to the induction and subsequent treatment for hypotension, an echocardiogram was performed. The patient was found to have an ejection fraction of 25%-30%. The anesthetic was uneventful for the remainder of the procedure, and following recovery, the patient was admitted by the heart failure team for further care.


Imaging intracellular fluorescent proteins at nanometer resolution.
We introduce a method for optically imaging intracellular proteins at nanometer spatial resolution. Numerous sparse subsets of photoactivatable fluorescent protein molecules were activated, localized (to approximately 2 to 25 nanometers), and then bleached. The aggregate position information from all subsets was then assembled into a superresolution image. We used this method--termed photoactivated localization microscopy--to image specific target proteins in thin sections of lysosomes and mitochondria; in fixed whole cells, we imaged vinculin at focal adhesions, actin within a lamellipodium, and the distribution of the retroviral protein Gag at the plasma membrane.


Lumbar lateral shift in a patient with interspinous device implantation: a case report.
Lumbar lateral shift (LLS) is a common clinical observation but has rarely been described in a patient with a history of lumbar surgery. The purpose of the current case report was to describe the use of the McKenzie Method of Mechanical Diagnosis and Therapy (MDT) in the multi-modal treatment of a patient with an LLS and a history of multiple surgical procedures in the lumbar spine, including interspinous process device (IPD) implantation. A 72-year-old female with chronic low back pain (LBP) and a surgical history in the lumbar spine was referred to physical therapy for radiating leg pain and presented with a right LLS. Her chief complaints included sitting for long periods, vacuuming and ascending stairs into her home. The patient was treated during eight visits over 30 days. Treatment interventions included manual shift correction, self-correction and management, joint mobilisation below the level of IPD implantation ,neurophysiology education, and development of a home exercise programme. At discharge, her leg pain was resolved and all goals had been met. The patient reported maintenance of gains at 6-month follow-up. Utilisation of the MDT approach, including LLS correction, produced positive outcomes in a complex patient with previous IPD implantation. Future research should investigate treatment and outcomes after invasive spinal procedures in similar patient populations to better inform clinical management. 4.


Young people's recall and perceptions of gambling advertising and intentions to gamble on sport.
There has been an increased international policy focus on the factors that may contribute to, and prevent, the normalization of gambling for young people. However, there is still limited research, which investigates the role of advertising in shaping young people's gambling attitudes and consumption intentions. Mixed methods study of 111 young people aged 11-16 years recruited from community basketball stadiums in Victoria, Australia, between May and July 2018. Interviewer-assisted surveys investigated recall and awareness of sports betting brands, perceptions of promotional strategies, intention to gamble, and reasons for betting on particular sports. Quantitative data were analyzed using descriptive statistics and χ2 tests. Thematic analyses were used to interpret qualitative responses. Young people had high recall and awareness of advertising, with most able to name at least one betting brand (n = 90, 81.1%), and many demonstrating a high awareness of the distinct characteristics (such as colors and appeal strategies) of different brands. A fifth of young people (n = 25, 22.5%) expressed intentions to gamble at 18 years, with boys significantly more likely than girls to state they would gamble (χ2 = 10.90, p = .001). Young people perceived that advertising strategies associated with inducement promotions would be the most influential in encouraging individuals to gamble. While many young people took promotions at face value, there was evidence that some were able to critically engage with and challenge the messages within marketing. Current regulatory structures appear to be ineffective in limiting young people's recall and awareness of gambling advertising. Lessons from tobacco control support the application of precautionary approaches as a more effective way to limit young people's development of positive gambling attitudes and behaviors.


Gene-Culture Coinheritance of a Behavioral Trait.
Many physical and behavioral traits in animals, including humans, are inherited both genetically and culturally. The presence of different inheritance systems affecting the same trait can result in complex evolutionary dynamics. Here, we present a general model that elucidates the distinct roles of cultural and genetic inheritance systems and their interaction in driving the evolution of complex phenotypes. In particular, we derive a Price equation that incorporates both cultural and genetic inheritance of a phenotype where the effects of genes and culture are additive. We then use this equation to investigate whether a genetically maladaptive phenotype can evolve under dual transmission. We examine the special case of altruism using an illustrative model and show that cultural selection can overcome genetic selection when the variance in culture is sufficiently high with respect to genes. We also show that the presence of cultural transmission can modify genetic selection itself, making genetic selection more favorable to a trait than under purely genetic inheritance. Last, we consider the effect of different timescales of genetic and cultural transmission. We discuss the implications of our results for understanding the evolution of important coinherited behaviors, including how our framework can be used to generate quantitative estimates of selection pressures required for a genetically maladaptive trait to evolve.


Imaging of beta-cell mass and function.
In both type 1 and type 2 diabetes mellitus, beta-cell mass (BCM), which exclusively produces insulin, is lost. Various therapeutic strategies are being developed that target BCM to restore its function by promoting beta-cell neogenesis and regeneration or by preventing its apoptosis. To this end, it is essential to identify biomarkers of BCM. Of the various imaging platforms, radionuclide-based imaging methods using radioligands that directly target BCM appear promising. In particular, the vesicular monoamine transporter type 2 (VMAT2), which is expressed almost exclusively by beta-cells and found in close association with insulin, can be noninvasively imaged with PET and (11)C-dihydrotetrabenazine or its derivatives. Despite the major limitation that beta-cells are low in abundance (1%-2%) and dispersed throughout the pancreas, VMAT2 PET is sensitive enough to detect VMAT2 signal and to allow kinetic model-based quantification of VMAT2 binding within the pancreas. However, these techniques are still in early stages, and careful further evaluations and technical developments are needed before they can be clinically used as a valid biomarker of BCM.


Construction and operation of microbial fuel cell with Chlorella vulgaris biocathode for electricity generation.
In this study, a modified microbial fuel cell (MFC) with a tubular photobioreactor (PHB) configuration as a cathode compartment was constructed by introducing Chlorella vulgaris to the cathode chamber used to generate oxygen in situ. Two types of cathode materials and light/dark cycles were used to test the effect on MFC with algae biocathode. Results showed that the use of algae is an effective approach because these organisms can act as efficient in situ oxygenators, thereby facilitating the cathodic reaction. Dissolved oxygen and voltage output displayed a clear light positive response and were drastically enhanced compared with the abiotic cathode. In particular, carbon paper-coated Pt used as a cathode electrode increased voltage output at a higher extent than carbon felt used as an electrode. The maximum power density of 24.4 mW/m2 was obtained from the MFC with algae biocathode which utilized the carbon paper-coated Pt as the cathode electrode under intermittent illumination. This density was 2.8 times higher than that of the abiotic cathode. Continuous illumination shortened the algal lifetime. These results demonstrated that intermittent illumination and cathode material-coated catalyst are beneficial to a more efficient and prolonged operation of MFC with C. vulgaris biocathode.