The empowerment paradox as a central challenge to patient centered medical home implementation in the veteran's health administration.
In this paper we present results of a mixed methods study conducted to identify barriers to team function among staff implementing patient aligned care teams - the Department of Veterans Affairs' patient centered medical home (PCMH) model. Using a convergent mixed methods design, we administered a standardized survey measure (Team and Individual Role Perception Survey) to assess work role challenge and engagement; and conducted discussion groups to gather context pertaining to role change. We found that the role of primary care providers is highly challenging and did not become less difficult over the initial year of implementation. Unexpectedly over the course of the first year nurse care managers reported a decrease in their perceptions of empowerment and clerical associates reported less skill variety. Qualitative data suggest that more skilled team members fail to delegate and share tasks within their teams. We characterize this interprofessional knowledge factor as an empowerment paradox where team members find it difficult to share tasks in ways that are counter to traditionally structured hierarchical roles. Health care systems seeking to implement PCMH should dedicate resources to facilitating within-team role knowledge and negotiation.


Tensor-based cortical surface morphometry via weighted spherical harmonic representation.
We present a new tensor-based morphometric framework that quantifies cortical shape variations using a local area element. The local area element is computed from the Riemannian metric tensors, which are obtained from the smooth functional parametrization of a cortical mesh. For the smooth parametrization, we have developed a novel weighted spherical harmonic (SPHARM) representation, which generalizes the traditional SPHARM as a special case. For a specific choice of weights, the weighted-SPHARM is shown to be the least squares approximation to the solution of an isotropic heat diffusion on a unit sphere. The main aims of this paper are to present the weighted-SPHARM and to show how it can be used in the tensor-based morphometry. As an illustration, the methodology has been applied in the problem of detecting abnormal cortical regions in the group of high functioning autistic subjects.


Area postrema and adjacent nucleus of the solitary tract in water and sodium balance.
Lesions of the area postrema (AP) and adjacent caudal medial nucleus of the solitary tract (cmNTS) cause significant changes in water and sodium balance. Lesioned rats display a permanent polydipsia, which in part is due to a primary polyuria. Water-to-food ratios are elevated chronically. Lesioned rats are unable to concentrate their urine as well as controls. In addition, lesioned rats overdrink in response to 24-h water deprivation. This lesion also causes a natriuresis and an overconsumption of 3% NaCl solution. These findings establish the AP-cmNTS as an important part of the neurocircuitry underlying water and sodium balance.


Mercury and human genotoxicity: critical considerations and possible molecular mechanisms.
Mercury compounds versatility explains their numerous applications in diverse areas of industry. The growing use of this metal has resulted in a significant increase of environment contamination and episodes of human intoxication, arousing the concern of international organisms. Meanwhile, consequences of these intoxication outbreaks are still not fully understood, especially if we consider long-term effects of chronic exposure to relatively low levels of mercury compounds. In the present manuscript, studies about the genotoxicity of mercury compounds, performed in vitro, in vivo, and/or including epidemiologic studies of human populations were reviewed. Some mercury compounds are known as teratogenic agents, especially affecting the normal development of the central nervous system; however, the connection between mercury exposure and carcinogenesis remains controversial. Since 1990s, epidemiological studies have begun to include an increasing number of human subjects, making the results more reliable: thus, increased genotoxicity was demonstrated in human populations exposed to mercury through diet, occupation or by carrying dental fillings. In fact, concentrations of methylmercury causing significant genotoxic alterations in vitro below both safety limit and concentration were associated with delayed psychomotor development with minimal signs of methylmercury poisoning. Based on mercury's known ability to bind sulfhydryl groups, several hypotheses were raised about potential molecular mechanisms for the metal genotoxicity. Mercury may be involved in four main processes that lead to genotoxicity: generation of free radicals and oxidative stress, action on microtubules, influence on DNA repair mechanisms and direct interaction with DNA molecules. All data reviewed here contributed to a better knowledge of the widespread concern about the safety limits of mercury exposure.


Sex determination from hand and foot dimensions in a North Indian population.
Hands and feet are often recovered from the site of natural as well as man-made disasters because of bomb blasts, train accidents, plane crashes, or mass homicides. This study is intended to establish standards for determination of sex from the dimensions of hands and feet in a North Indian population. The data for this study comprise 123 men and 123 women aged between 17 and 20 years from the "Rajput" population of Himachal Pradesh in North India. Four anthropometric measurements viz. hand length, hand breadth, foot length, and foot breadth have been taken on both sides of each subject following international anthropometric standards. The hand index (hand breadth/hand length × 100) and the foot index (foot breadth/foot length × 100) were calculated. Sectioning points and regression models are derived for the hand and foot dimensions and the derived indices. The hand and foot dimensions show a higher accuracy in sex determination by sectioning point analysis when compared to hand and foot index. Of the hand and the foot dimensions, hand breadth and foot breadth showed better accuracy in sex determination. Hand index and foot index remain poor sex discriminators in the study.


Heterosynaptic plasticity prevents runaway synaptic dynamics.
Spike timing-dependent plasticity (STDP) and other conventional Hebbian-type plasticity rules are prone to produce runaway dynamics of synaptic weights. Once potentiated, a synapse would have higher probability to lead to spikes and thus to be further potentiated, but once depressed, a synapse would tend to be further depressed. The runaway synaptic dynamics can be prevented by precisely balancing STDP rules for potentiation and depression; however, experimental evidence shows a great variety of potentiation and depression windows and magnitudes. Here we show that modifications of synapses to layer 2/3 pyramidal neurons from rat visual and auditory cortices in slices can be induced by intracellular tetanization: bursts of postsynaptic spikes without presynaptic stimulation. Induction of these heterosynaptic changes depended on the rise of intracellular calcium, and their direction and magnitude correlated with initial state of release mechanisms. We suggest that this type of plasticity serves as a mechanism that stabilizes the distribution of synaptic weights and prevents their runaway dynamics. To test this hypothesis, we develop a cortical neuron model implementing both homosynaptic (STDP) and heterosynaptic plasticity with properties matching the experimental data. We find that heterosynaptic plasticity effectively prevented runaway dynamics for the tested range of STDP and input parameters. Synaptic weights, although shifted from the original, remained normally distributed and nonsaturated. Our study presents a biophysically constrained model of how the interaction of different forms of plasticity--Hebbian and heterosynaptic--may prevent runaway synaptic dynamics and keep synaptic weights unsaturated and thus capable of further plastic changes and formation of new memories.


Anxiety, depression and psychological well-being in a cohort of South African adults with Type 2 diabetes mellitus.
The prevalence of diabetes mellitus (DM) has increased at alarming rates globally. South Africa has the second highest number of people in Africa living with DM, with prevalence rates being among the top five countries in Africa. Accordingly, psychological issues associated with DM have been a growing focus of attention. Studies have found that patients with DM have elevated levels of anxiety and depression, and decreased levels of well-being. In South Africa, there is a paucity of studies on the psychological issues associated with DM. The aim of this paper was to explore the prevalence and association of anxiety, depressive features and psychological well-being in patients with Type 2 DM. In a cross-sectional survey, patients with Type 2 DM were recruited from public and private facilities. The Hospital Anxiety and Depression Scale (HADS), the General Health Questionnaire (GHQ-28) and WHO-5 Well-being Index (WHO-5) were administered. Four hundred and one participants completed the questionnaires. On the WHO-5, 277 (69%) reported good well-being, while 124 (31%) indicated poor well-being and were considered at risk for depressive features. On the HADS, 186 (46%) had mild-to-severe depressive features and 128 (32%) had mild-to-severe anxiety. There was a strong negative correlation between the WHO-5, HADS and General Health Questionnaire (GHQ) scales, which indicated that an increase in anxiety and depressive features decreased psychological well-being. Health-care providers should identify and treat anxiety and depression as a standard part of diabetes care. Patients should also be referred to the appropriate mental health professional as part of the management of diabetes.


A HER2-binding Affibody molecule labelled with 68Ga for PET imaging: direct in vivo comparison with the 111In-labelled analogue.
Overexpression of HER2 receptors is a prognostic and predictive biomarker in breast cancer and a number of other malignancies. Radionuclide molecular imaging of HER2 overexpression may influence patient management making treatment more personalized. Earlier, (111)In-DOTA-Z(HER2:342-pep2) (ABY-002) Affibody molecule demonstrated excellent imaging of HER2-expressing xenografts in mice shortly after injection. The use of the positron-emitting nuclide (68)Ga instead of (111)In might increase both the sensitivity of HER2 imaging and accuracy of expression quantification. The goal of this study was to prepare and characterize (68)Ga-labelled ABY-002. (68)Ga labelling of ABY-002 was optimized. In vitro cell binding and procession of (68)Ga-ABY-002 was evaluated. Biodistribution and tumour targeting of (68)Ga-ABY-002 and (111)In-ABY-002 was compared in vivo by paired-label experiments. ABY-002 was incubated with (68)Ga at 90 degrees C for 10 min resulting in a radiochemical labelling yield of over 95%. Capacity for specific binding to HER2-expressing cells was retained. In vivo, both (68)Ga-ABY-002 and (111)In-ABY-002 demonstrated specific targeting of SKOV-3 xenografts and high-contrast imaging. Background radioactivity in blood, lungs, gastrointestinal tract and muscle fell more rapidly for (68)Ga-ABY-002 compared with (111)In-ABY-002 favouring imaging shortly after injection. For (68)Ga-ABY-002, a tumour uptake of 12.4 +/- 3.8%ID/g and a tumour to blood ratio of 31 +/- 13 were achieved at 2 h post-injection. (68)Ga-ABY-002 is easy to label and provides high-contrast imaging within 2 h after injection. This makes it a promising candidate for clinical molecular imaging of HER2 expression in malignant tumours.


Anionic bipyridyl ligands for applications in metallasupramolecular chemistry.
The facile synthesis of anionic bipyridyl ligands with dinuclear clathrochelate cores is described. These metalloligands can be obtained in high yields by the reactions of M(ClO4 )2 (H2 O)6 (M: Zn, Mn, or Co) with 4-pyridylboronic acid and 2,6-diformyl-4-methylphenol oxime or 2,6-diformyl-4-tert-butylphenol oxime, followed by deprotonation. The ligands are interesting building blocks for metallasupramolecular chemistry, as evidenced by the formation of a Pt-based molecular square and four coordination polymers with 2D or 3D network structures. Competition experiments reveal that the utilization of anionic bipyridyl ligands can result in significantly more stable assemblies.


Proinflammatory and proosteoclastogenic potential of peripheral blood mononuclear cells from Gaucher patients: Implication for bone pathology.
Gaucher disease (GD) is caused by mutations in the GBA gene that confer a deficient level of activity of glucocerebrosidase (GCase). This deficiency leads to the accumulation of the glycolipid glucocerebroside in the lysosomes of cells of monocyte/macrophage system. Bone compromise in Gaucher disease patients is the most disabling aspect of the disease. However, pathophysiological aspects of skeletal alterations are still poorly understood. On the other hand it is well known that inflammation is a key player in GD pathology. In this work, we revealed increased levels of the proinflammatory CD14(+)CD16(+) monocyte subset and increased inflammatory cytokine production by monocytes and T cells in the circulation of GD patients. We showed increased levels of osteoclast precursors in PBMC from patients and a higher expression of RANKL in the surface of T cells. PBMC from patients presented higher osteoclast differentiation compared to healthy controls when cultured in the presence of M-CSF alone or in combination with RANKL. In vitro treatment with Velaglucerase reduced osteoclast levels to control levels. On the other hand THP-1 derived osteoclast precursors cultured in the presence of conditioned media from PBMC of GD patients presented higher differentiation to active osteoclasts. This induction involved TNF-α and RANKL.


Access to abortion under the heath exception: a comparative analysis in three countries.
Despite Britain, Colombia, and some Mexican states sharing a health exception within their abortion laws, access to abortion under the health exception varies widely. This study examines factors that result in heterogeneous application of similar health exception laws and consequences for access to legal abortion. Our research adds to previous literature by comparing implementation of similar abortion laws across countries to identify strategies for full implementation of the health exception. We conducted a cross-country comparative descriptive study synthesizing data from document and literature review, official abortion statistics, and interviews with key informants. We gathered information on the use and interpretation of the health exception in the three countries from peer-reviewed literature, court documents, and grey literature. We next extracted public and private abortion statistics to understand the application of the law in each setting. We used a matrix to synthesize information and identify key factors in the use of the law. We conducted in-depth interviews with doctors and experts familiar with the health exception laws in each country and analyzed the qualitative data based on the previously identified factors. The health exception is used broadly in Britain, somewhat in Colombia, and very rarely in Mexican states. We identified five factors as particularly salient to application of the health exception in each setting: 1) comprehensiveness of the law including explicit mention of mental health, 2) a strong public health sector that funds abortion, 3) knowledge of and attitudes toward the health exception law, including guidelines for physicians in providing abortion, 4) dissemination of information about the health exception law, and 5) a history of court cases that protect women and clarify the health exception law. The health exception is a valuable tool for expanding access to legal abortion. Differences in the use of the health exception as an indication for legal abortion result in wide access for women in Britain to nearly no access in Mexican states. Our findings highlight the difference between theoretical and real access to legal abortion. The interpretation and application of the health exception law are pivotal to expanding real access to abortion.


To have and not to have sex: When multiple evolutions of conditional use of sex elegantly solve the question in the ant genus Cataglyphis.
Organisms use an amazingly large diversity of mechanisms to pass on their genes to the next generation. Sex is ancestral in eukaryotes, where it remains the most widespread way of reproduction. By combining one's genes with those of a partner, sex entails a dilution of one's genes at each generation. Evolution has been particularly creative in devising mechanisms allowing females to avoid this dilution, from classical parthenogenesis to the elimination of male genes after fertilization (Bell, 1982). Moreover, the term parthenogenesis includes various forms. Parthenogenesis can be used for female (thelytoky) or male (arrhenotoky) production and it can be associated with different cytological mechanisms, from strict clonality to meiotic division with the fusion of two of the four products of meiosis to restore diploidy (Suomalainen, Saura, & Lokki, 1987). Understanding the evolution of these diverse reproductive systems remains one of the most exciting and longstanding questions in evolutionary biology. By characterizing the reproductive systems of 11 species from the thermophilic ant genus Cataglyphis, in this issue of Molecular Ecology, Kuhn, Darras, Paknia, and Aron (2020) show the high lability of parthenogenesis, with multiple independent evolution of facultative thelytoky from sexual ancestors. The diversity of life history traits and social characteristics of this genus (e.g., mode of colony foundation, female polyandry) provides a unique and exciting opportunity to investigate the social and environmental factors driving the evolution of reproductive systems in social Hymenoptera.


Decoding the temporal nature of brain GR activity in the NFκB signal transition leading to depressive-like behavior.
The fine-tuning of neuroinflammation is crucial for brain homeostasis as well as its immune response. The transcription factor, nuclear factor-κ-B (NFκB) is a key inflammatory player that is antagonized via anti-inflammatory actions exerted by the glucocorticoid receptor (GR). However, technical limitations have restricted our understanding of how GR is involved in the dynamics of NFκB in vivo. In this study, we used an improved lentiviral-based reporter to elucidate the time course of NFκB and GR activities during behavioral changes from sickness to depression induced by a systemic lipopolysaccharide challenge. The trajectory of NFκB activity established a behavioral basis for the NFκB signal transition involved in three phases, sickness-early-phase, normal-middle-phase, and depressive-like-late-phase. The temporal shift in brain GR activity was differentially involved in the transition of NFκB signals during the normal and depressive-like phases. The middle-phase GR effectively inhibited NFκB in a glucocorticoid-dependent manner, but the late-phase GR had no inhibitory action. Furthermore, we revealed the cryptic role of basal GR activity in the early NFκB signal transition, as evidenced by the fact that blocking GR activity with RU486 led to early depressive-like episodes through the emergence of the brain NFκB activity. These results highlight the inhibitory action of GR on NFκB by the basal and activated hypothalamic-pituitary-adrenal (HPA)-axis during body-to-brain inflammatory spread, providing clues about molecular mechanisms underlying systemic inflammation caused by such as COVID-19 infection, leading to depression.


In Vitro Differentiation of Human CD4+FOXP3+ Induced Regulatory T Cells (iTregs) from Naïve CD4+ T Cells Using a TGF-β-containing Protocol.
Regulatory T cells (Tregs) are an integral part of peripheral tolerance, suppressing immune reactions against self-structures and thus preventing autoimmune diseases. Clinical approaches to adoptively transfer Tregs, or to deplete Tregs in cancer, are underway with promising first outcomes. Because the number of naturally occurring Tregs (nTregs) is very limited, studying certain Treg features using in vitro induced Tregs (iTregs) can be advantageous. To date, the best although not absolutely specific protein marker to delineate Tregs is the transcription factor FOXP3. Despite the importance of Tregs including non-redundant roles of peripherally induced Tregs, the protocols to generate iTregs are currently controversial, particularly for human cells. This protocol therefore describes the in vitro differentiation of human CD4+FOXP3+ iTregs from human naïve T cells using a range of Treg-inducing factors (TGF-β plus IL-2 only, or their combination with retinoic acid, rapamycin or butyrate) in parallel. It also describes the phenotyping of these cells by flow cytometry and qRT-PCR. These protocols result in reproducible expression of FOXP3 and other Treg signature genes and enable the study of general FOXP3-regulatory mechanisms as well as protocol-specific effects to delineate the impact of certain factors. iTregs can be utilized to study various phenotypic aspects as well as molecular mechanisms of Treg induction. Detailed molecular studies are facilitated by relatively large cell numbers that can be obtained. A limitation for the application of iTregs is the relative instability of FOXP3 expression in these cells compared to nTregs. iTregs generated by these protocols can also be used for functional assays such as studying their suppressive function, in which iTregs induced by TGF-β plus retinoic acid and rapamycin display superior suppressive activity. However, the suppressive capacity of iTregs can differ from nTregs and the use of appropriate controls is crucial.


High sensitivity detection of salivary 8-hydroxy deoxyguanosine levels in patients with chronic periodontitis.
Inflammation is associated with hydroxyl radical damage to DNA as a result of oxidative stress. 8-Hydroxy deoxyguanosine (8-OHdG) is a marker of this process and its levels in saliva could be linked to the severity of periodontal inflammation. The aim of this study was to test the sensitivity of liquid chromatography with tandem mass spectrometry (LC-MS/MS) in comparison to enzyme-linked immune sorbent assay (ELISA) for the detection of 8-OHdG in saliva in patients with chronic periodontitis before and after periodontal treatment. Saliva samples were collected from 23 patients (eight females and 15 males; 46.1 ± 5.1 years of age) with generalized chronic periodontitis and 25 (15 females and 10 males; 44.9 ± 6.8 years of age) periodontally healthy individuals. Patients received initial periodontal treatment consisting of scaling and root planing and were evaluated at baseline and after 6 wk of completion of non-surgical therapy. Salivary 8-OHdG levels were measured using ELISA and LC-MS/MS before and after the treatment. Clinically, plaque index, gingival index, clinical attachment level, bleeding on probing, gingival recession and probing pocket depth were measured at baseline and after 6 wk. Salivary levels of 8-OHdG decreased significantly after the non-surgical periodontal treatment (p < 0.001). Statistically significant positive correlations were observed between plaque index, gingival index, probing pocket depth, clinical attachment level, bleeding on probing values and LC-MS/MS and ELISA levels of 8-OHdG (p < 0.001). LC-MS/MS is a reliable and sensitive method for evaluating salivary 8-OHdG levels to monitor the treatment response of periodontitis.


Prevalence and natural history of arachnoid cysts in adults.
Arachnoid cysts are a frequent finding on intracranial imaging. The prevalence and natural history of these cysts in adults are not well defined. We retrospectively reviewed the electronic medical records of a consecutive series of adults who underwent brain MRI over a 12-year interval to identify those with arachnoid cysts. The MRI studies were reviewed to confirm the diagnosis. For those patients with arachnoid cysts, we evaluated presenting symptoms, cyst size, and cyst location. Patients with more than 6 months' clinical and imaging follow-up were included in a natural history analysis. A total of 48,417 patients underwent brain MRI over the study period. Arachnoid cysts were identified in 661 patients (1.4%). Men had a higher prevalence than women (p < 0.0001). Multiple arachnoid cysts occurred in 30 patients. The most common locations were middle fossa (34%), retrocerebellar (33%), and convexity (14%). Middle fossa cysts were predominantly left-sided (70%, p < 0.001). Thirty-five patients were considered symptomatic and 24 underwent surgical treatment. Sellar and suprasellar cysts were more likely to be considered symptomatic (p < 0.0001). Middle fossa cysts were less likely to be considered symptomatic (p = 0.01. The criteria for natural history analysis were met in 203 patients with a total of 213 cysts. After a mean follow-up of 3.8 ± 2.8 years (for this subgroup), 5 cysts (2.3%) increased in size and 2 cysts decreased in size (0.9%). Only 2 patients developed new or worsening symptoms over the follow-up period. Arachnoid cysts are a common incidental finding on intracranial imaging in all age groups. Although arachnoid cysts are symptomatic in a small number of patients, they are associated with a benign natural history for those presenting without symptoms.


Metal-on-Metal Bearing: Is This the End of the Line? We Do Not Think So.
Recent studies have recommended the discontinuation of metal-on-metal (MoM) components in total hip arthroplasty (THA) because of adverse effects reported with large-diameter MoM THA. This is despite favorable long-term results observed with 28 and 32 mm MoM bearings. The aim of this study was to assess the value of calls for an end to MoM bearings as THA components. Specifically, we wish to address the risks associated with MoM bearings including adverse soft tissue reactions, metal ion release, and carcinogenic risk. The study evaluates the arguments in the literature reporting on MoM (adverse soft tissue reactions, metal ion release, and carcinogenic risk) and the experience of the current authors who re-introduced these bearings in 1995. They are balanced by a benefit-risk review of the literature and the authors' experience with MoM use. Adverse reactions to metallic debris as well as metal ion release are predictable and can be prevented by adequate design (arc of coverage, clearance), metallurgy (forged instead of cast alloy, high-carbide content), and appropriate component orientation. There is no scientific evidence that carcinogenicity is increased in subjects with MoM hip prostheses. MoM articulations appear to be attractive allowing safe hip resurfacing, decreasing the risk of THA revision in active patients, and providing secure THA fixation with cement in cages in severely deformed hips. MoM bearings in women of child-bearing age are controversial, but long-term data on metallic devices in adolescents undergoing spinal surgery seem reassuring. Adequate selection of MoM articulations ensures their safe use. These articulations are sensitive to orientation. Fifteen years of safe experience with 28- and 32-mm bearings of forged alloy and high-carbide content is the main reason for retaining them in primary and revision THA.


Specific recognition of ZNF217 and other zinc finger proteins at a surface groove of C-terminal binding proteins.
Numerous transcription factors recruit C-terminal binding protein (CtBP) corepressors. We show that the large zinc finger protein ZNF217 contacts CtBP. ZNF217 is encoded by an oncogene frequently amplified in tumors. ZNF217 contains a typical Pro-X-Asp-Leu-Ser (PXDLS) motif that binds in CtBP's PXDLS-binding cleft. However, ZNF217 also contains a second motif, Arg-Arg-Thr (RRT), that binds a separate surface on CtBP. The crystal structure of CtBP bound to an RRTGAPPAL peptide shows that it contacts a surface crevice distinct from the PXDLS binding cleft. Interestingly, both PXDLS and RRT motifs are also found in other zinc finger proteins, such as RIZ. Finally, we show that ZNF217 represses several promoters, including one from a known CtBP target gene, and mutations preventing ZNF217's contact with CtBP reduce repression. These results identify a new CtBP interaction motif and establish ZNF217 as a transcriptional repressor protein that functions, at least in part, by associating with CtBP.


Insights of Patients and Clinicians on the Promise of the Experience Sampling Method for Psychiatric Care.
This qualitative study aimed to map the relevance of the experience sampling method (ESM) for psychiatric practice and identify barriers and facilitators for implementation, as perceived by patients and clinicians. Participants were 22 patients with various diagnoses and 21 clinicians (e.g., psychiatrists, psychologists) who participated in interviews or focus groups. Using Atlas.TI, the authors conducted qualitative thematic analysis to analyze the transcripts, resulting in four themes: applications, advantages, undesirable effects, and requirements for implementation of ESM in care. Clinicians and patients believed ESM could be relevant in every phase of care to increase patients' awareness, insight, and self-management; personalize interventions; and alert patients to rising symptoms. Further, ESM was expected to improve the patient-clinician relationship; lead to objective, personalized, reliable and visual data; and increase efficiency of care. However, participants warned against high assessment burden and potential symptom worsening. This study provides first evidence that the potential of ESM is recognized by both patients and clinicians. Key recommendations for optimal implementation of ESM in psychiatric care include flexible application of ESM, collaboration between patient and clinician, regular evaluation, awareness of negative reactivity, availability to patients with different psychiatric syndromes, and implementation by an interdisciplinary team of patients, clinicians, researchers, and information technology specialists.


A bacterial antibiotic resistance accelerator and applications.
The systematic emergence of drug resistance remains a major problem in the treatment of infectious diseases (antibiotics) and cancer (chemotherapy), with possible common fundamental origins linking bacterial antibiotic resistance and emergence of chemotherapy resistance. The common link may be evolution in a complex fitness landscape with connected small population niches. We report a detailed method for observing bacterial adaptive behavior in heterogeneous microfluidic environment designed to mimic the environmental heterogeneity found in natural microbial niches. First, the device is structured with multiple connected micro-chambers that allow the cell population to communicate and organize into smaller populations. Second, bacteria evolve within an antibiotic gradient generated throughout the micro-chambers that creates a wide range of fitness landscapes. High-resolution images of the adaptive response to the antibiotic stress are captured by epifluorescence microscopy at various levels of the bacterial organization for quantitative analysis. Thus, the experimental setup we have developed provides a powerful frame for visualizing evolution at work: bacterial movement, survival and death. It also presents a basis for exploring the rates at which drug resistance arises in bacteria and other biological contexts such as cancer.


Differential expression of nuclear matrix proteins during the differentiation of human neuroblastoma SK-N-SH cells induced by retinoic acid.
To investigate the alteration of nuclear matrix proteins (NMPs) during the differentiation of neuroblastoma SK-N-SH cells induced by retinoic acid (RA), differentiation markers were detected by immunocytochemistry and NMPs were selectively extracted and subjected to two-dimensional gel electrophoresis analysis. Immunocytochemical observation demonstrated that the expression of neuronal markers was up-regulated in SK-N-SH cells following RA treatment. Meanwhile, 52 NMPs (41 of which were identified) changed significantly during SK-N-SH differentiation; four of these NMPs were further confirmed by immunoblotting. This study suggests that the differentiation of neuroblastoma cells was accompanied by the altered expression of neuronal markers and NMPs. The presence of some differentially expressed NMPs was related to the proliferation and differentiation of neuroblastomas. Our results may help to reveal the relationship between NMPs and neuroblastoma carcinogenesis and reversion, as well as elucidate the regulatory principals driving neural cell proliferation and differentiation.


Cannabidiol (CBD) enhances lipopolysaccharide (LPS)-induced pulmonary inflammation in C57BL/6 mice.
Cannabidiol (CBD) is a plant-derived cannabinoid that has been predominantly characterized as anti-inflammatory. However, it is clear that immune effects of cannabinoids can vary with cannabinoid concentration, or type or magnitude of immune stimulus. The present studies demonstrate that oral administration of CBD enhanced lipopolysaccharide (LPS)-induced pulmonary inflammation in C57BL/6 mice. The enhanced inflammatory cell infiltrate as observed in bronchoalveolar lavage fluid (BALF) was comprised mainly of neutrophils, with some monocytes. Concomitantly, CBD enhanced pro-inflammatory cytokine mRNA production, including tumor necrosis factor-α (Tnfa), interleukins (IL)-5 and -23 (Il6, Il23), and granulocyte colony stimulating factor (Gcsf). These results demonstrate that the CBD-mediated enhancement of LPS-induced pulmonary inflammation is mediated at the level of transcription of a variety of pro-inflammatory genes. The significance of these studies is that CBD is part of a therapeutic currently in use for spasticity and pain in multiple sclerosis patients, and therefore it is important to further understand mechanisms by which CBD alters immune function.


Clinical course of ectopic pregnancy: A single-center experience.
The objective was to highlight the frequency, clinical profile, and predisposing factors of ectopic pregnancy (EP) in a general hospital. This descriptive study was conducted at the Obstetrics and Gynaecology department of Hera General hospital, Makkah, Saudi Arabia, from July 1, 2009 to December 29, 2010. Data were collected on chief medical complaints, sociodemographic characteristics, past obstetrics and gynecological history, management done, and outcome of management. Data were analyzed using Microsoft Office Excel (version 2007). Out of total 7564 pregnancies, 44 (0.58%) patients were diagnosed as EP. Out of 44, 22 (50%) patients presented within 24 h of onset of symptoms. Mean age was 28 ± 7 years. Multigravida were predominant in 25 (57%), and 21 (48%) had gestational age of 6-8 weeks at the time of presentation; the common presenting features were amenorrhea (41, 93.2%), abdominal pain (39, 88.6%), and tenderness (38, 86%). Previous pelvic surgery (13, 29.5%), infertility treatment (11, 25%), and pelvic inflammatory disease (10, 22.7%) were the common predisposing factors. Twenty-five (57%) presented with ruptured EP and were operated within 24 h, and the remaining were kept under observation till further diagnosis. After confirming the diagnosis, 12/19 underwent laparoscopy, whereas 7/19 received medical treatment. Surgery confirmed fallopian tube pregnancies in 35 (94.5%). No mortality was observed. Previous pelvic surgeries were the major etiological factor for EP. Other factors were infertility treatment and pelvic inflammatory disease. The most common site of EP was fallopian tubes.


Multiple improvement of astaxanthin biosynthesis in Xanthophyllomyces dendrorhous by a combination of conventional mutagenesis and metabolic pathway engineering.
Xanthophyllomyces dendrorhous (Phaffia rhodozyma) is the only yeast or fungus that synthesizes the commercially attractive carotenoid astaxanthin. For a suitable bioprocess, the wild type has to be modified for increasing biomass content. To achieve this, a two step strategy has been followed. At first, random mutagenesis was applied leading to colonies with substantially higher astaxanthin content. Then, the resulting strain was genetically engineered by targeting limiting reactions for further enhancement of astaxanthin biosynthesis. This combinatorial approach together with selection of an appropriate growth medium resulted in highest astaxanthin biomass contents reported to date for X. dendrorhous. In a fermenter culture, its maximum content was 9.7 mg/g dry weight.


Melanocortin peptide therapy for the treatment of arthritic pathologies.
Arthritic pathologies are a major cause of morbidity within the western world, with rheumatoid arthritis affecting approximately 1% of adults. This review highlights the therapeutic potential of naturally occurring hormones and their peptides, in both arthritic models of disease and patients. The arthritides represent a group of closely related pathologies in which cytokines, joint destruction, and leukocytes play a causal role. Here we discuss the role of naturally occurring pro-opiomelanocortin (POMC)-derived melanocortin peptides (e.g., alpha melanocyte stimulating hormone [alpha-MSH]) and synthetic derivatives in these diseases. Melanocortins exhibit their biological efficacy by modulating proinflammatory cytokines and subsequent leukocyte extravasation. Their biological effects are mediated via seven transmembrane G-protein-coupled receptors, of which five have been cloned, identified, and termed MC1 to MC5. Adrenocorticotrophic hormone represents the parent molecule of the melanocortins; the first 13 amino acids of which (termed alpha-MSH) have been shown to be the most pharmacologically active region of the parent hormone. The melanocortin peptides have been shown to display potent anti-inflammatory effects in both animal models of disease and patients. The potential anti-inflammatory role for endogenous peptides in arthritic pathologies is in its infancy. The ability to inhibit leukocyte migration, release of cytokines, and induction of anti-inflammatory proteins appears to play an important role in affording protection in arthritic injury, and thus may lead to potential therapeutic targets.


Remarkable recovery in an infant presenting with extensive perinatal cervical cord injury.
Cervical-cord damage is a complication of a difficult delivery, and results in spinal shock with flaccidity progressing to spastic paralysis. Conventionally, outlook for such patients is extremely poor and most will recover only slightly from quadriplegia and autonomic dysfunction. Here, we report a case in which the extent of damage considerably contrasted with the outcome and recovery. A full-term baby girl born by difficult vaginal delivery displayed bilateral flaccid paralysis of the lower limbs with absent spontaneous movements, weakness of both upper limbs, hyporeflexia in all limbs and axial hypotonia. MRI of cervicothoracic spine exhibited raised signal intensity in the dorsal aspects of C7 to T1 signifying myelopathy. MRI at 4 months revealed a near-total transection of the cervical cord. However, at 6 months, the child could move all lower limbs independently with a marked increase in power. There was no spasticity, wasting or incontinence. Reflexes had also returned.


The Climate Response to Emissions Reductions Due to COVID-19: Initial Results From CovidMIP.
Many nations responded to the corona virus disease-2019 (COVID-19) pandemic by restricting travel and other activities during 2020, resulting in temporarily reduced emissions of CO2, other greenhouse gases and ozone and aerosol precursors. We present the initial results from a coordinated Intercomparison, CovidMIP, of Earth system model simulations which assess the impact on climate of these emissions reductions. 12 models performed multiple initial-condition ensembles to produce over 300 simulations spanning both initial condition and model structural uncertainty. We find model consensus on reduced aerosol amounts (particularly over southern and eastern Asia) and associated increases in surface shortwave radiation levels. However, any impact on near-surface temperature or rainfall during 2020-2024 is extremely small and is not detectable in this initial analysis. Regional analyses on a finer scale, and closer attention to extremes (especially linked to changes in atmospheric composition and air quality) are required to test the impact of COVID-19-related emission reductions on near-term climate.


Does antibiotic prophylaxis at implant placement decrease early implant failures? A Cochrane systematic review.
Marco Esposito is the first author of two of the included studies; however, he was not involved in the quality assessment of these trials. This review is based on a Cochrane systematic review entitled 'Interventions for replacing missing teeth: antibiotics at dental implant placement to prevent complications' published in The Cochrane Library (see http://www.cochrane.org for more information). Cochrane systematic reviews are regularly updated to include new research, and in response to comments and criticisms from readers. If you wish to comment on this review, please send your comments to the Cochrane website or to Marco Esposito. The Cochrane Library should be consulted for the most recent version of the review. The results of a Cochrane Review can be interpreted differently, depending on people's perspectives and circumstances. Please consider the conclusions presented carefully. They are the opinions of the review authors, and are not necessarily shared by the Cochrane Collaboration. To assess the beneficial or harmful effects of systemic prophylactic antibiotics at dental implant placement versus no antibiotic/placebo administration and, if antibiotics are of benefit, to find which type, dosage and duration is the most effective. The Cochrane Oral Health Group's Trials Register, the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE and EMBASE were searched up to 2 June 2010 for randomised controlled clinical trials (RCTs) with a follow-up of at least 3 months comparing the administration of various prophylactic antibiotic regimens versus no antibiotics to patients undergoing dental implant placement. Outcome measures were prosthesis failures, implant failures, postoperative infections and adverse events (gastrointestinal, hypersensitivity, etc.). Screening of eligible studies, assessment of the methodological quality of the trials and data extraction were conducted in duplicate and independently by two review authors. Meta-analyses were conducted. Four RCTs were identified: three comparing 2 g of preoperative amoxicillin versus placebo (927 patients) and the other comparing 1 g of preoperative amoxicillin plus 500 mg four times a day for 2 days versus no antibiotics (80 patients). The meta-analyses of the four trials showed a statistically significantly higher number of patients experiencing implant failures in the group not receiving antibiotics: risk ratio=0.40 (95% confidence interval (CI) 0.19 to 0.84). The number needed to treat (NNT) to prevent one patient having an implant failure is 33 (95% CI 17-100), based on a patient implant failure rate of 5% in patients not receiving antibiotics. The other outcomes were not statistically significant, and only two minor adverse events were recorded, one in the placebo group. There is some evidence suggesting that 2 g of amoxicillin given orally 1 h preoperatively significantly reduce failures of dental implants placed in ordinary conditions. No significant adverse events were reported. It might be sensible to suggest the use of a single dose of 2 g prophylactic amoxicillin prior to dental implant placement. It is still unknown whether post-operative antibiotics are beneficial, and which is the most effective antibiotic.


Job Strain as a Risk Factor for Peripheral Artery Disease: A Multi-Cohort Study.
Background Job strain is implicated in many atherosclerotic diseases, but its role in peripheral artery disease (PAD) is unclear. We investigated the association of job strain with hospital records of PAD, using individual-level data from 11 prospective cohort studies from Finland, Sweden, Denmark, and the United Kingdom. Methods and Results Job strain (high demands and low control at work) was self-reported at baseline (1985-2008). PAD records were ascertained from national hospitalization data. We used Cox regression to examine the associations of job strain with PAD in each study, and combined the study-specific estimates in random effects meta-analyses. We used τ2, I2, and subgroup analyses to examine heterogeneity. Of the 139 132 participants with no previous hospitalization with PAD, 32 489 (23.4%) reported job strain at baseline. During 1 718 132 person-years at risk (mean follow-up 12.8 years), 667 individuals had a hospital record of PAD (3.88 per 10 000 person-years). Job strain was associated with a 1.41-fold (95% CI, 1.11-1.80) increased average risk of hospitalization with PAD. The study-specific estimates were moderately heterogeneous (τ2=0.0427, I2: 26.9%). Despite variation in their magnitude, the estimates were consistent in both sexes, across the socioeconomic hierarchy and by baseline smoking status. Additional adjustment for baseline diabetes mellitus did not change the direction or magnitude of the observed associations. Conclusions Job strain was associated with small but consistent increase in the risk of hospitalization with PAD, with the relative risks on par with those for coronary heart disease and ischemic stroke.


A framework for teaching medical students and residents about practice-based learning and improvement, synthesized from a literature review.
To create a framework for teaching the knowledge and skills of practice-based learning and improvement to medical students and residents based on proven, effective strategies. The authors conducted a Medline search of English-language articles published between 1996 and May 2001, using the term "quality improvement" (QI), and cross-matched it with "medical education" and "health professions education." A thematic-synthesis method of review was used to compile the information from the articles. Based on the literature review, an expert panel recommended educational objectives for practice-based learning and improvement. Twenty-seven articles met the inclusion criteria. The majority of studies were conducted in academic medical centers and medical schools and 40% addressed experiential learning of QI. More than 75% were qualitative case reports capturing educational outcomes, and 7% included an experimental study design. The expert panel integrated data from the literature review with the Dreyfus model of professional skill acquisition, the Institute for Healthcare Improvement's (IHI) knowledge domains for improving health care, and the ACGME competencies and generated a framework of core educational objectives about teaching practice-based learning and improvement to medical students and residents. Teaching the knowledge and skills of practice-based learning and improvement to medical students and residents is a necessary and important foundation for improving patient care. The authors present a framework of learning objectives-informed by the literature and synthesized by the expert panel-to assist educational leaders when integrating these objectives into a curriculum. This framework serves as a blueprint to bridge the gap between current knowledge and future practice needs.


'A caring professional attitude': What service users and carers seek in graduate nurses and the challenge for educators.
With the publication of the new NMC standards for pre-registration nursing education, undergraduate curricula are being written in universities across England. There are many drivers for the curricula but one that has until recently received scant attention is the service user's and carer's voice. This paper discusses the findings of a qualitative study that asked 52 service users and carers about the qualities they sought in nurses and their views on nurse education. Eight focus groups were conducted with a broad range of service users and carers from primary and secondary care, and voluntary organisations. Data were analysed using the framework approach facilitated by a qualitative analysis software programme. The sample was diverse, but there were similarities in the qualities they valued in nurses. They sought technical competence, knowledge and willingness to seek information, but overwhelmingly prioritised 'a caring professional attitude'. This was articulated as empathy, communication skills and non-judgmental patient centred care: major themes in the new NMC standards. Our participants also expressed concern about whether the educational preparation of nurses can develop these caring qualities. We discuss this concern, the challenges for nurse educators it presents and how we can engage service users and carers in shaping and delivering our new curricula.


Specific and common changes in Nicotiana benthamiana gene expression in response to infection by enveloped viruses.
Microarrays derived from Solanum tuberosum expressed sequence tags were used to test the hypothesis that genetically distinct enveloped viruses elicit unique changes in Nicotiana benthamiana gene expression. The results of our study, which included Sonchus yellow net virus (SYNV), a plant rhabdovirus that replicates in the nucleus of infected cells, and Impatiens necrotic spot virus (INSV), a plant bunyavirus that replicates in the cytoplasm, were consistent with this hypothesis. Statistically significant changes (P< or =0.01) in the expression of 275, 2646 and 4165 genes were detected in response to INSV at 2, 4 and 5 days post-inoculation (d.p.i.), respectively. In contrast, 35, 665 and 1458 genes were expressed differentially in response to SYNV at 5, 11 and 14 d.p.i., respectively. The microarray results were verified by Northern hybridization using a subset of these genes as probes. Notably, INSV, but not SYNV, induced expression of small heat-shock protein genes to high levels. In contrast to SYNV, infection by INSV resulted in downregulation of all histone genes, of which the downregulation of histone 2b expression to very low levels was confirmed by Northern hybridization. The expression of a putative WRKY transcription factor at 11 d.p.i., but not at 5 or 14 d.p.i., in SYNV-infected tissue suggested that the temporal response to virus infection was identified readily using our experimental design. Overall, infection by INSV resulted in larger fold changes in host gene expression relative to infection by SYNV. Taken together, the present data demonstrate differential responses of a common host to two genetically distinct viruses.


Differential gene expression in insects: transcriptional control.
Studies on transcriptional control of gene expression play a pivotal role in many areas of biology. In non-Drosophilid insects, the cuticle, chorion, immune response, silk gland, storage proteins, and vitellogenin are foci for advances in basic research on promoter elements and transcription factors. Insects offer other advantages for gene regulation studies, including the availability of applied problems. In non-Drosophilid insects, the most serious problem for transcriptional control studies is the lack of homologous in vivo expression systems. Once this deficiency is addressed, the full impact of research on transcription control will be realized throughout the field of entomology.


Growth in utero, blood pressure in childhood and adult life, and mortality from cardiovascular disease.
In national samples of 9921 10 year olds and 3259 adults in Britain systolic blood pressure was inversely related to birth weight. The association was independent of gestational age and may therefore be attributed to reduced fetal growth. This suggests that the intrauterine environment influences blood pressure during adult life. It is further evidence that the geographical differences in average blood pressure and mortality from cardiovascular disease in Britain partly reflect past differences in the intrauterine environment. Within England and Wales 10 year olds living in areas with high cardiovascular mortality were shorter and had higher resting pulse rates than those living in other areas. Their mothers were also shorter and had higher diastolic blood pressures. This suggests that there are persisting geographical differences in the childhood environment that predispose to differences in cardiovascular mortality.


Prediction of expected years of life using whole-genome markers.
Genetic factors are believed to account for 25% of the interindividual differences in Years of Life (YL) among humans. However, the genetic loci that have thus far been found to be associated with YL explain a very small proportion of the expected genetic variation in this trait, perhaps reflecting the complexity of the trait and the limitations of traditional association studies when applied to traits affected by a large number of small-effect genes. Using data from the Framingham Heart Study and statistical methods borrowed largely from the field of animal genetics (whole-genome prediction, WGP), we developed a WGP model for the study of YL and evaluated the extent to which thousands of genetic variants across the genome examined simultaneously can be used to predict interindividual differences in YL. We find that a sizable proportion of differences in YL--which were unexplained by age at entry, sex, smoking and BMI--can be accounted for and predicted using WGP methods. The contribution of genomic information to prediction accuracy was even higher than that of smoking and body mass index (BMI) combined; two predictors that are considered among the most important life-shortening factors. We evaluated the impacts of familial relationships and population structure (as described by the first two marker-derived principal components) and concluded that in our dataset population structure explained partially, but not fully the gains in prediction accuracy obtained with WGP. Further inspection of prediction accuracies by age at death indicated that most of the gains in predictive ability achieved with WGP were due to the increased accuracy of prediction of early mortality, perhaps reflecting the ability of WGP to capture differences in genetic risk to deadly diseases such as cancer, which are most often responsible for early mortality in our sample.


Socioeconomic differences in the burden of disease in Sweden.
We sought to analyse how much of the total burden of disease in Sweden, measured in disability-adjusted life years (DALYs), is a result of inequalities in health between socioeconomic groups. We also sought to determine how this unequal burden is distributed across different disease groups and socioeconomic groups. Our analysis used data from the Swedish Burden of Disease Study. We studied all Swedish men and women in three age groups (15-44, 45-64, 65-84) and five major socioeconomic groups. The 18 disease and injury groups that contributed to 65% of the total burden of disease were analysed using attributable fractions and the slope index of inequality and the relative index of inequality. About 30% of the burden of disease among women and 37% of the burden among men is a differential burden resulting from socioeconomic inequalities in health. A large part of this unequally distributed burden falls on unskilled manual workers. The largest contributors to inequalities in health for women are ischaemic heart disease, depression and neurosis, and stroke. For men, the largest contributors are ischaemic heart disease, alcohol addiction and self-inflicted injuries. This is the first study to use socioeconomic differences, measured by socioeconomic position, to assess the burden of disease using DALYs. We found that in Sweden one-third of the burden of the diseases we studied is unequally distributed. Studies of socioeconomic inequalities in the burden of disease that take both mortality and morbidity into account can help policy-makers understand the magnitude of inequalities in health for different disease groups.


Appearance of the hymen in newborns.
This study was undertaken to evaluate the genital anatomy of female newborns. Hymenal configuration and the number and location of clefts, bumps, tags, and ridges were observed in 468 neonates; photographs for confirmation were obtained for 449 neonates (96%). Hymenal configurations observed included an annular hymen with a central or ventrally displaced orifice in 80% (372/468) of neonates, a fimbriated hymen in 19% (90/468), and a septated or cribriform hymen in 1% (6/468). Configurations significantly differed by race (P less than .001) but no difference was noted by gestational age or weight. Clefts, which occurred only on the ventral 180 degrees of the rim, were observed in 34% of neonates with an annular hymen. Fifty-six percent of neonates had a longitudinal intravaginal ridge, 87% had an external hymenal ridge, and 13% had a tag extending from the rim or a ridge. Periurethral ligaments were frequently observed. It is concluded that lateral and ventral clefts, intravaginal and external ridges, and periurethral bands are normal anatomical findings in the newborn.


NLRX1 protein attenuates inflammatory responses to infection by interfering with the RIG-I-MAVS and TRAF6-NF-κB signaling pathways.
The nucleotide-binding domain and leucine-rich-repeat-containing (NLR) proteins regulate innate immunity. Although the positive regulatory impact of NLRs is clear, their inhibitory roles are not well defined. We showed that Nlrx1(-/-) mice exhibited increased expression of antiviral signaling molecules IFN-β, STAT2, OAS1, and IL-6 after influenza virus infection. Consistent with increased inflammation, Nlrx1(-/-) mice exhibited marked morbidity and histopathology. Infection of these mice with an influenza strain that carries a mutated NS-1 protein, which normally prevents IFN induction by interaction with RNA and the intracellular RNA sensor RIG-I, further exacerbated IL-6 and type I IFN signaling. NLRX1 also weakened cytokine responses to the 2009 H1N1 pandemic influenza virus in human cells. Mechanistically, Nlrx1 deletion led to constitutive interaction of MAVS and RIG-I. Additionally, an inhibitory function is identified for NLRX1 during LPS activation of macrophages where the MAVS-RIG-I pathway was not involved. NLRX1 interacts with TRAF6 and inhibits NF-κB activation. Thus, NLRX1 functions as a checkpoint of overzealous inflammation.


Viral chemotherapy.
New antiviral compounds are being tested constantly and may be of considerable value with increasing availability. More than 200 analogues of purines and pyrimidines have been found to inhibit DNA and RNA viruses. Adenine arabinoside is most effective against disseminated herpes simplex virus and disseminated herpes zoster. Idoxuridine is useful in treatment of herpetic keratitis. Interferon still is in the experimental stage, and, because of its short half-life and high cost, it probably will not be released in the near future. Amantadine appears to be useful in prevention of A2 influenza, but its value against swine flu has not been established. Methisazone is effective in prevention of smallpox and in the treatment of complications of vaccinia.


Biotransformation of sevoflurane by rat neonate liver slices.
Sevoflurane [CF3-CH(OCH2F)-CF3] is biotransformed to inorganic fluoride (F-) and hexafluoroisopropanol, which forms a glucuronide conjugate. Although sevoflurane may be used in newborns without fully developed biotransformation activity, studies were performed using liver slices from rat neonates to determine sevoflurane disposition. Sevoflurane was vaporized in sealed roller culture vials to produce a continuous saturating dose (0.5 mM). After incubation, slices and incubation media were sonicated and centrifuged to remove debris. The supernatant fraction was analyzed for F-, hexafluoroisopropanol, and hexafluoroisopropanol-glucuronide conjugate. The metabolism of sevoflurane by liver slices increased proportionately with time with a stoichiometric production (1:1) of hexafluoroisopropanol and F- in all age groups. Only glucuronide conjugates of hexafluoroisopropanol were found. The rate of sevoflurane biotransformation measured as fluoride production was similar among slices prepared from all neonate age groups. Although no hexafluoroisopropanol-glucuronide was generated by slices from 4-, 6-, and 8-day-old neonates, by day 21, 17% of the total hexafluoroisopropanol is glucuronidated. This contrasts with the lower levels of free hexafluoroisopropanol typically seen in adults liver slices, wherein 51% of the hexafluoroisopropanol was glucuronidated. These studies indicate that sevoflurane is equally metabolized to hexafluoroisopropanol and F-, but a deficiency in glucuronosyltransferase occurs in neonates.


Ultrasonography and electrophysiological study of median nerve in patients with essential tremor.
Essential tremor (ET) is a common movement disorder characterized by postural or kinetic tremor. We aimed to evaluate median nerve enlargement in patients with ET using ultrasonography (USG). Thirty-eight hands from 19 patients with ET and 24 hands from 13 controls underwent nerve conduction studies (NCS) and USG at the wrist. Tremor severity was measured using the Fahn-Tolosa-Marin Tremor Rating Scale (FTM-TRS). The median nerve cross sectional area (mCSA) in USG and NCS parameters were compared using ANCOVA. We evaluated the correlation between mCSA and NCS parameters or FTM-TRS scores using linear regression analysis. mCSA was significantly larger (p<0.001) and NCS parameters were different in two groups. Also, mCSA was negatively correlated with part B and C scores of FTM-TRS (p<0.001 and p = 0.039, respectively). In conclusion, median nerve enlargement with the changes of NCS parameters was observed and correlated with the severity of tremor in patients with ET.


Targeting the EMT transcription factor TWIST1 overcomes resistance to EGFR inhibitors in EGFR-mutant non-small-cell lung cancer.
Patients with EGFR-mutant non-small-cell lung cancer (NSCLC) have significantly benefited from the use of EGFR tyrosine kinase inhibitors (TKIs). However, long-term efficacy of these therapies is limited due to de novo resistance (~30%) as well as acquired resistance. Epithelial-mesenchymal transition transcription factors (EMT-TFs), have been identified as drivers of EMT-mediated resistance to EGFR TKIs, however, strategies to target EMT-TFs are lacking. As the third generation EGFR TKI, osimertinib, has now been adopted in the first-line setting, the frequency of T790M mutations will significantly decrease in the acquired resistance setting. Previously less common mechanisms of acquired resistance to first generation EGFR TKIs including EMT are now being observed at an increased frequency after osimertinib. Importantly, there are no other FDA approved targeted therapies after progression on osimertinib. Here, we investigated a novel strategy to overcome EGFR TKI resistance through targeting the EMT-TF, TWIST1, in EGFR-mutant NSCLC. We demonstrated that genetic silencing of TWIST1 or treatment with the TWIST1 inhibitor, harmine, resulted in growth inhibition and apoptosis in EGFR-mutant NSCLC. TWIST1 overexpression resulted in erlotinib and osimertinib resistance in EGFR-mutant NSCLC cells. Conversely, genetic and pharmacological inhibition of TWIST1 in EGFR TKI-resistant EGFR-mutant cells increased sensitivity to EGFR TKIs. TWIST1-mediated EGFR TKI resistance was due in part to TWIST1 suppression of transcription of the pro-apoptotic BH3-only gene, BCL2L11 (BIM), by directly binding to BCL2L11 intronic regions and promoter. As such, pan-BCL2 inhibitor treatment overcame TWIST1-mediated EGFR TKI resistance and were more effective in the setting of TWIST1 overexpression. Finally, in a mouse model of autochthonous EGFR-mutant lung cancer, Twist1 overexpression resulted in erlotinib resistance and suppression of erlotinib-induced apoptosis. These studies establish TWIST1 as a driver of resistance to EGFR TKIs and provide rationale for use of TWIST1 inhibitors or BCL2 inhibitors as means to overcome EMT-mediated resistance to EGFR TKIs.


Prolonged institutional rearing is associated with atypically large amygdala volume and difficulties in emotion regulation.
Early adversity, for example poor caregiving, can have profound effects on emotional development. Orphanage rearing, even in the best circumstances, lies outside of the bounds of a species-typical caregiving environment. The long-term effects of this early adversity on the neurobiological development associated with socio-emotional behaviors are not well understood. Seventy-eight children, who include those who have experienced orphanage care and a comparison group, were assessed. Magnetic resonance imaging (MRI) was used to measure volumes of whole brain and limbic structures (e.g. amygdala, hippocampus). Emotion regulation was assessed with an emotional go-nogo paradigm, and anxiety and internalizing behaviors were assessed using the Screen for Child Anxiety Related Emotional Disorders, the Child Behavior Checklist, and a structured clinical interview. Late adoption was associated with larger corrected amygdala volumes, poorer emotion regulation, and increased anxiety. Although more than 50% of the children who experienced orphanage rearing met criteria for a psychiatric disorder, with a third having an anxiety disorder, the group differences observed in amygdala volume were not driven by the presence of an anxiety disorder. The findings are consistent with previous reports describing negative effects of prolonged orphanage care on emotional behavior and with animal models that show long-term changes in the amygdala and emotional behavior following early postnatal stress. These changes in limbic circuitry may underlie residual emotional and social problems experienced by children who have been internationally adopted.


Mechanisms of human cytomegalovirus persistence and latency.
Human cytomegalovirus (HCMV) is a ubiquitous beta-herpesvirus that causes severe disease primarily in immunosuppressed individuals. A major characteristic of HCMV with obvious clinical importance is the ability of the virus to establish lifelong infection within the host following the initial acute infection. One strategy used by HCMV to maintain itself within the host is the establishment of cellular sites of persistent infection and viral latency. Recent studies have identified endothelial cells and monocyte-derived macrophages (MDM) as sites of HCMV persistence and latency. These studies show that endothelial cell origin and MDM differentiation pathway are critical factors that influence the characteristics of HCMV replication in these cell types. The specific HCMV genes involved in endothelial cell and MDM tropism are unknown. However, studies in the closely related murine cytomegalovirus (MCMV) model have provided considerable insight into viral genes that enable replication in these cell types. This review will focus on mechanisms of HCMV replication in endothelial cells and MDM, and on the viral genes involved in regulation of viral replication in these important cell types.


SRY, SOX9, and DAX1 expression patterns during human sex determination and gonadal development.
SRY, SOX9, and DAX1 are key genes in human sex determination, by virtue of their associated male-to-female sex reversal phenotypes when mutated (SRY, SOX9) or over-expressed (DAX1). During human sex determination, SRY is expressed in 46,XY gonads coincident with sex cord formation, but also persists as nuclear protein within Sertoli cells at 18 weeks gestation. High-level SOX9 expression in the sex cords of the testis parallels that seen during mouse development, however in humans, SOX9 transcripts also are detected in the developing ovary. Low-level DAX1 expression predates peak SRY expression by at least 10 days, and persists in Sertoli cells throughout the entire sex determination period. In Dosage Sensitive Sex reversal, the anti-testis properties of DAX1 over-expression could act prior to the peak effects of SRY and continue during the period of SOX9 expression. These findings highlight expression differences for the SRY, SOX9, and DAX1 genes during sex determination in humans and mice. These results provide a direct framework for future investigation into the mechanisms underlying normal and abnormal human sex determination.


NADPH oxidase mediates oxidative stress in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine model of Parkinson's disease.
Parkinson's disease (PD) is a neurodegenerative disorder of uncertain pathogenesis characterized by a loss of substantia nigra pars compacta (SNpc) dopaminergic (DA) neurons, and can be modeled by the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Both inflammatory processes and oxidative stress may contribute to MPTP- and PD-related neurodegeneration. However, whether inflammation may cause oxidative damage in MPTP and PD is unknown. Here we show that NADPH-oxidase, the main reactive oxygen species (ROS)-producing enzyme during inflammation, is up-regulated in SNpc of human PD and MPTP mice. These changes coincide with the local production of ROS, microglial activation, and DA neuronal loss seen after MPTP injections. Mutant mice defective in NADPH-oxidase exhibit less SNpc DA neuronal loss and protein oxidation than their WT littermates after MPTP injections. We show that extracellular ROS are a main determinant in inflammation-mediated DA neurotoxicity in the MPTP model of PD. This study supports a critical role for NADPH-oxidase in the pathogenesis of PD and suggests that targeting this enzyme or enhancing extracellular antioxidants may provide novel therapies for PD.


A life course approach to mortality in Mexico.
Research on early life socioeconomic status (SES), education and mortality is less established in developing countries. This analysis aims to determine how SES and education are patterned across the life course and associated with adult mortality in Mexico. Data comes from 2001-2012 Mexican Health & Aging Study (Mexican adults age 50+, n= 11,222). Cox proportional hazard models predict mortality using baseline covariates. In unadjusted analyses, similar mortality was seen across levels of early life SES. Lower early life SES was associated with better survival after accounting for education in the younger cohort. Lower education was only associated with mortality in the younger cohort. Early life SES was associated with education but the relationship between education and mortality differed across cohorts in Mexico. Selective survival and differential returns to education may explain differences.


Particle-swarm optimization of broadband nanoplasmonic arrays.
We used the particle swarm optimization algorithm, an evolutionary computational technique, to design metal nanoparticle arrays that produce broadband plasmonic field enhancement over the entire visible spectral range. The resulting structures turn out to be aperiodic and feature dense Fourier spectra with many closely packed particle clusters. We conclude that broadband field-enhancement effects in nanoplasmonics can be achieved by engineering aperiodic arrays with a large number of spatial frequencies that provide the necessary interplay between long-range diffractive interactions at multiple length scales and near-field quasi-static coupling within small nanoparticle clusters.


Automated detection of fecal contamination of apples by multispectral laser-induced fluorescence imaging.
Animal feces are a suspected source of contamination of apples by disease-causing organisms such as Echerichia coli O157. Laser-induced fluorescence was used to detect different amounts of feces from dairy cows, deer, and a dairy pasture applied to Red Delicious apples. One day after application, detection for 1:2 and 1:20 dilutions was nearly 100%, and for 1:200 dilutions (<15 ng of dry matter) detection was >80%. Detection after apples had been washed and brushed was lowest for pasture feces; detection for 1:2, 1:20, and 1:200 dilutions of feces was 100%, 30%, and 0%, respectively. This technology may encourage development of commercial systems for detecting fecal contamination of apples.


Predicting new solids and superconductors.
It is now possible to start with a simple model of a solid composed of atomic cores and itinerant valence electrons and compute the total energy for a given structural arrangement of atoms with enough precision to predict the existence of new solids and their properties. The application of the model based on the pseudopotential method is described with silicon chosen as a prototype material. With only information about the constituent atoms, the electronic, structural, vibrational, and even superconducting properties of solids can be calculated from first principles. The successful predictions of superconductivity in highly condensed hexagonal silicon and the existence of new high-pressure semiconductor phases are highlighted. A discussion is presented of the use of the method to discover new stable or metastable solids at high pressures.


Laser fabrication of Au nanorod aggregates microstructures assisted by two-photon polymerization.
We demonstrate fabrication of Au nanorod aggregates microstructures by means of a femtosecond near-infrared laser. The laser light was tightly focused into colloidal Au nanorods dispersed in photopolymerizable metyl-methacrylate (MMA) compound to induce two-photon polymerization (TPP). TPP of MMA glued the nanorods together to form solid microstrucures of aggregates. The laser light excited a local surface plasmon, resulting in confinement of TPP in the vicinity of nanorods. Concurrenly occurring optical accumulation of nanorods created a unique mechanism for the formation of nanorod aggregates into desired microstructures. This technique would be a clue for a novel micro/nanofabrication method for plasmonic materials and devices.


Diversity and distribution of nematodes associated with terrestrial slugs in the Western Cape Province of South Africa.
A survey of nematodes associated with native and introduced species of terrestrial slugs was conducted in the Western Cape Province of South Africa, in order to gather new data regarding diversity and distribution. A total of 521 terrestrial slugs were collected from 35 localities throughout the Western Cape. All slugs were dissected and examined for the presence of internal nematodes. Extracted nematodes were identified using a combination of molecular (18S rRNA gene sequencing) and morphological techniques. Nematodes were found parasitizing slugs at 14 of the 35 sites examined, amounting to 40% of sample sites. Of all slugs, 6% were infected with nematodes. A total of seven species of nematode were identified in the province, including Agfa flexilis, Angiostoma sp., Phasmarhabditis sp. SA1, Phasmarhabditis sp. SA2, Caenorhabditis elegans, Panagrolaimus sp. and Rhabditis sp. Of these species, four were thought to be parasitic to slugs (A. flexilis, Angiostoma sp., Phasmarhabditis sp. SA1 and Phasmarhabditis sp. SA2), as opposed to forming necromenic or phoretic associations. Three new species of slug-parasitic nematode were identified during this study (Angiostoma sp., Phasmarhabditis sp. SA1 and Phasmarhabditis sp. SA2).


Pathogen Presence in European Starlings Inhabiting Commercial Piggeries in South Australia.
The majority of bacterial diarrhea-causing illnesses in domestic pigs result from infection with Escherichia coli, Salmonella spp., or Campylobacter spp. These bacterial enteropathogens also correspond with the most-common bacteria isolated from wild birds. Additionally, viral pathogens such as avian influenza virus (AIV), West Nile virus (WNV, including Kunjin disease), and Newcastle disease virus (NDV) may also be carried and transmitted by birds in Australia. Introduced European starlings (Sturnus vulgarus) are one of the most-frequently reported birds on piggeries in Australia. The presence of the three bacterial pathogens, Salmonella spp., Campylobacter spp., and Escherichia coli , as well as the three viral pathogens AIV, WNV, and NDV, were evaluated in starlings captured on four commercial piggeries in South Australia. A total of 473 starlings were captured on the four piggeries in 2008 and 2009. A cloacal swab was taken from each bird and cultured for bacterial identification, with follow-up serotyping of any positives, whilst fifty samples were analyzed by PCR for the three target viral pathogens. There was no AIV, WNV, or NDV detected in the 50 starlings sampled. Escherichia coli was found to be present in the starling populations on all four piggeries whilst Salmonella spp. and Campylobacter jejuni were found to be present only in the starling population sampled on one piggery. Serotyping identified pig-pathogenic strains of the bacteria. The prevalence of these production-limiting bacterial pathogens in starlings, coupled with the large starling populations often found inside piggeries during daylight hours in the summer months, presents a disease transmission risk and jeopardizes piggery disease management. Removal of starlings from agricultural enterprises (as shown by international studies), or prevention of starling access to animal feed and water, could substantially reduce the risk of transmission of enterobacterial pathogens from starlings to livestock.


An intensive behavioral weight loss intervention and hot flushes in women.
Higher body mass index is associated with worse hot flushes during menopause but the effect of weight loss on flushing is unclear. Self-administered questionnaires were used to assess bothersome hot flushes in a 6-month randomized controlled trial of an intensive behavioral weight loss program (intervention) vs a structured health education program (control) in 338 women who were overweight or obese and had urinary incontinence. Weight, body mass index, abdominal circumference, physical activity, calorie intake, blood pressure, and physical and mental functioning were assessed at baseline and at 6 months. Repeated-measures proportional odds models examined intervention effects on bothersome hot flushes and potential mediating factors. Approximately half of participants (n = 154) were at least slightly bothered by hot flushes at baseline. Among these women, the intervention was associated with greater improvement in bothersome flushes vs control (odds ratio [OR] for improvement by 1 Likert category, 2.25; 95% confidence interval [CI], 1.20-4.21). Reductions in weight (OR, 1.32; 95% CI, 1.08-1.61; per 5-kg decrease), body mass index (1.17; 1.05-1.30; per 1-point decrease), and abdominal circumference (1.32; 1.07-1.64; per 5-cm decrease) were each associated with improvement in flushing, but changes in physical activity, calorie intake, blood pressure, and physical and mental functioning were not related. The effect of the intervention on flushing was modestly diminished after adjustment for multiple potential mediators (OR, 1.92; 95% CI, 0.95-3.89). Among women who were overweight or obese and had bothersome hot flushes, an intensive behavioral weight loss intervention resulted in improvement in flushing relative to control. Trial Registration clinicaltrials.gov Identifier: NCT00091988.


Psychobehavioural factors are more strongly associated with successful weight management than predetermined satiety effect or other characteristics of diet.
This study aimed to investigate factors associated with weight management, especially whether satiety value of food as a part of a weight-maintenance diet would affect self-regulation of food intake and weight management. Altogether 82 obese subjects completed the study consisting of weight-loss and weight-maintenance (WM) periods. During the WM, subjects were randomized into higher- and lower-satiety food groups. No differences were observed in the changes in body weight, energy intake, or eating behaviour between the groups, even despite the different macronutrient compositions of the diets. However, when regarding all study subjects, success in WM was most strongly associated with a greater increase in the flexible control of eating and experience of greater easiness of WM and control of food intake and a greater decrease in uncontrollable eating and psychological distress. Psychobehavioural factors seem to be more strongly associated with successful weight management than the predetermined satiety effect or other characteristics of the diet.


Dynamical configuration of binary near-Earth asteroid (66391) 1999 KW4.
Dynamical simulations of the coupled rotational and orbital dynamics of binary near-Earth asteroid 66391 (1999 KW4) suggest that it is excited as a result of perturbations from the Sun during perihelion passages. Excitation of the mutual orbit will stimulate complex fluctuations in the orbit and rotation of both components, inducing the attitude of the smaller component to have large variation within some orbits and to hardly vary within others. The primary's proximity to its rotational stability limit suggests an origin from spin-up and disruption of a loosely bound precursor within the past million years.


RNA m6A methylation across the transcriptome.
Since the early days of foundational studies of nucleic acids, many chemical moieties have been discovered to decorate RNA and DNA in diverse organisms. In mammalian cells, one of these chemical modifications, N6-methyl adenosine (m6A), is unique in a way that it is highly abundant not only on RNA polymerase II (RNAPII) transcribed, protein-coding transcripts but also on non-coding RNAs, such as ribosomal RNAs and snRNAs, mediated by distinct, evolutionarily conserved enzymes. Here, we review RNA m6A modification in the light of the recent appreciation of nuclear roles for m6A in regulating chromatin states and gene expression, as well as the recent discoveries of the evolutionarily conserved methyltransferases, which catalyze methylation of adenosine on diverse sets of RNAs. Considering that the substrates of these enzymes are involved in many important biological processes, this modification warrants further research to understand the molecular mechanisms and functions of m6A in health and disease.


Direct electrical control of IgG conformation and functional activity at surfaces.
We have devised a supramolecular edifice involving His-tagged protein A and antibodies to yield surface immobilized, uniformly oriented, IgG-type, antibody layers with Fab fragments exposed off an electrode surface. We demonstrate here that we can affect the conformation of IgGs, likely pushing/pulling electrostatically Fab fragments towards/from the electrode surface. A potential difference between electrode and solution acts on IgGs' charged aminoacids modulating the accessibility of the specific recognition regions of Fab fragments by antigens in solution. Consequently, antibody-antigen affinity is affected by the sign of the applied potential: a positive potential enables an effective capture of antigens; a negative one pulls the fragments towards the electrode, where steric hindrance caused by neighboring molecules largely hampers the capture of antigens. Different experimental techniques (electrochemical quartz crystal microbalance, electrochemical impedance spectroscopy, fluorescence confocal microscopy and electrochemical atomic force spectroscopy) were used to evaluate binding kinetics, surface coverage, effect of the applied electric field on IgGs, and role of charged residues on the phenomenon described. These findings expand the concept of electrical control of biological reactions and can be used to gate electrically specific recognition reactions with impact in biosensors, bioactuators, smart biodevices, nanomedicine, and fundamental studies related to chemical reaction kinetics.


Blazed high-efficiency x-ray diffraction via transmission through arrays of nanometer-scale mirrors.
Diffraction gratings are ubiquitous wavelength dispersive elements for photons as well as for subatomic particles, atoms, and large molecules. They serve as enabling devices for spectroscopy, microscopy, and interferometry in numerous applications across the physical sciences. Transmission gratings are required in applications that demand high alignment and figure error tolerances, low weight and size, or a straight-through zero-order beam. However, photons or particles are often strongly absorbed upon transmission, e.g., in the increasingly important extreme ultraviolet (EUV) and soft x-ray band, leading to low diffraction efficiency. We demonstrate the performance of a critical-angle transmission (CAT) grating in the EUV and soft x-ray band that for the first time combines the advantages of transmission gratings with the superior broadband efficiency of blazed reflection gratings via reflection from nanofabricated periodic arrays of atomically smooth nanometer-thin silicon mirrors at angles below the critical angle for total external reflection. The efficiency of the CAT grating design is not limited to photons, but also opens the door to new, sensitive, and compact experiments and applications in atom and neutron optics, as well as for the efficient diffraction of electrons, ions, or molecules.


Prolactin and Thyroid Stimulating Hormone (TSH) Levels and Sexual Dysfunction in Patients with Schizophrenia Treated with Conventional Antipsychotic Medication: A Cross-Sectional Study.
BACKGROUND This study aimed to investigate the relationship between serum profiles of prolactin and thyroid stimulating hormone (TSH) and sexual dysfunction in patients with schizophrenia treated with conventional antipsychotic medication. MATERIAL AND METHODS A hospital-based cross-sectional study included 118 patients, age range 18-57 years (55 men, 63 women), with a confirmed diagnosis of schizophrenia. All patients were stable after antipsychotic treatment. Serum levels of hormones, including prolactin, follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), progesterone, testosterone, thyroid-stimulating hormone (TSH), triiodothyronine (T3), thyroxine (T4), free triiodothyronine (FT3) and free thyroxine (FT4), were detected in venous blood. The Positive and Negative Syndrome Scale (PANSS) score was used to measure symptom severity of patients with schizophrenia. The Mandarin Chinese version of the Arizona Sexual Experience Scale (ASEX), a 5-item scale, was used to measure sexual function. RESULTS There were 66 patients (55.9%) who had hyperprolactinemia, the prevalence of hyperprolactinemia was markedly higher in the sexual dysfunction group than the non-sexual dysfunction group (91.8% vs. 17.5%) (P<0.001). Mean prolactin levels were significantly increased in patients with sexual dysfunction compared with the patients without sexual dysfunction (P<0.001), with a higher incidence in female patients. Subclinical hypothyroidism and hyperprolactinemia were found to be independently associated with sexual dysfunction, and an increased PANSS negative score was an independent risk factor for the development of sexual dysfunction. CONCLUSIONS The incidence of sexual dysfunction was significantly increased in patients with schizophrenia. Hyperprolactinemia and subclinical hypothyroidism were associated with sexual dysfunction, especially in female patients.


Global Thrombosis Test - a possible monitoring system for the effects and safety of dabigatran.
Dabigatran is an alternative to warfarin (WF) for the thromboprophylaxis of stroke in patients with non-valvular atrial fibrillation (NVAF). The advantage of dabigatran over WF is that monitoring is not required; however, a method to monitor the effect and the safety of dabigatran is not currently available. The Global Thrombosis Test (GTT) is a novel method to assess both clot formation and lysis activities under physiological conditions. The aim of this study was to evaluate whether treatment with dabigatran might affect shear-induced thrombi (occlusion time [OT], sec) by the GTT, and to investigate the possibility that the GTT could be useful as a monitoring system for dabigatran. The study population consisted of 50 volunteers and 43 NVAF patients on WF therapy, who were subsequently switched to dabigatran. Using the GTT, the thrombotic status was assessed one day before and 1 month after switching anticoagulation from WF to dabigatran. The OT was 524.9 ± 17.0 sec in volunteers whereas that of NVAF patients on WF therapy was 581.7 ± 26.3 sec. The switch from WF to dabigatran significantly prolonged OT (784.5 ± 19.3 sec). One patient on WF therapy and 12 patients on dabigatran therapy were shown to have OT > 900 sec. The GTT could be used to assess the risk of dabigatran-related bleeding complications.


Preparation of highly anisotropic cobalt ferrite/silica microellipsoids using an external magnetic field.
Magnetic cobalt ferrite/silica microparticles having both an original morphology and an anisotropic nanostructure are synthesized through the use of an external magnetic field and nanoparticles characterized by a high magnetic anisotropy. The association of these two factors implies that the ESE (emulsion and solvent evaporation) sol-gel method employed here allows the preparation of silica microellipsoids containing magnetic nanoparticles aggregated in large chains. It is clearly shown that without this combination, microspheres characterized by an isotropic distribution of the magnetic nanoparticles are obtained. While the chaining of the cobalt ferrite nanoparticles inside the silica matrix is related to the increase of their magnetic dipolar interactions, the ellipsoidal shape of the microparticles may be explained by the elongation of the sol droplets in the direction of the external magnetic field during the synthesis. Because of their highly anisotropic structure, these microparticles exhibit permanent magnetic moments, which are responsible, at a larger scale, for the existence of strong magnetic dipolar interactions. Therefore, when they are dispersed in water, the microellipsoids self-assemble into large and irregular chains. These interactions can be reinforced by the use of external magnetic field, allowing the preparation of very large permanent chains. This research illustrates how nanostructured particles exhibiting complex architectures can be elaborated through simple, fast, and low-cost methods, such as the use of external fields in combination with soft chemistry.


Tolerated doses in zebrafish of cytochalasins and jasplakinolide for comparison with tolerated doses in mice in the evaluation of pre-clinical activity of microfilament-directed agents in tumor model systems in vivo.
Chemotherapeutic approaches involving microtubule-directed agents such as the vinca alkaloids and taxanes are used extensively and effectively in clinical cancer therapy. There is abundant evidence of critical cytoskeletal differences involving microfilaments between normal and neoplastic cells, and a variety of natural products and semi-synthetic derivatives are available to exploit these differences in vitro. In spite of the availability of such potential anti-neoplastic agents, there has yet to be an effective microfilament-directed agent approved for clinical use. Cytochalasins are mycogenic toxins derived from a variety of fungal sources that have shown promising in vitro efficacy in disrupting microfilaments and producing remarkable cell enlargement and multi-nucleation in cancer cells without producing enlargement and multi-nucleation in normal blood cells. Jasplakinolide is a sponge toxin that stabilizes and rigidifies microfilaments. Insufficient in vivo data has been acquired to determine whether any of the microfilament-directed agents have valuable preferential anticancer activity in pre-clinical tumor model systems. This is partly because the limited availability of these agents precludes their initial use in large-scale mammalian pre-clinical studies. Therefore, the present study sought to determine the tolerated in vivo doses of cytochalasins and jasplakinolide in zebrafish (Danio rerio), a well-studied fish cancer model that is 1.5% the size of mice. We also determined the tolerated levels of a variety of clinically active anti-neoplastic agents in zebrafish for comparison with tolerated murine doses as a means to allow comparison of toxicities in zebrafish expressed as μM concentrations with toxicities in mice expressed in mg/kg. Tolerated doses in zebrafish with various cytochalasins or jasplakinolide were determined by adding the solubilized test agent to water in which the fish were maintained for 24 h, then restored to their normal tanks and monitored for a total of 96 h. Cytochalasin D at 0.2 μM gave an approximate LD50 in zebrafish, while cytochalasin B was fully-tolerated at 5 μM, and gave an LD50 of 10 μM. 21,22-dihydrocytochalasin B was fully-tolerated at 10 μM. Cytochalasin C was tolerated fully at 1 μM, ten-fold higher than the level for cytochalasin D that was tolerated. Jasplakinolide at 0.5 μM did not exhibit any apparent acute toxicity or affect fish behavior for four days, but delayed toxicity was evident at days 4 and 6 when the fish died. Further, the addition of 5 μM glutathione (GSH) at the time of treatment substantially decreased the toxicity of 10 μM cytochalasin B, a level of cytochalasin B that not otherwise tolerated in vivo. Such observations were likely due to GSH-mediated alkylation of C-20 in cytochalasin B, thereby reducing the rate of oxidation to the highly toxic congener, cytochalasin A, and reacting with any cytochalazin A formed. The protective effects of GSH are further supported by its ability to react with α, β-unsaturated ketone moieties, as is found in cytochalasin A. GSH at 0.8 uM was able to reduce the toxicity of 0.8 μM cytochalasin D, but it took 20 μM GSH to fully protect against the toxicity of 0.8 μM cytochalasin D. Pre-clinical evaluation of rare natural products such as microfilamented-directed agents for efficacy in vivo in tumor-bearing zebrafish is a feasible prospect. Dose-limiting toxicities in zebrafish expressed as μM concentrations in water can be used to estimate in vivo toxicities in mice expressed as mg/kg.


Clinical features and structured clinical evaluation of vasculitis.
Systemic vasculitides are a group of heterogeneous conditions with overlapping patterns of clinical and laboratory manifestations. Moreover, clinical features can be non-specific and seemingly disparate. A major factor in defining optimal therapy and measuring treatment response is careful disease assessment targeting four main domains: activity, damage, prognosis and quality of life/function. Assessment tools such as the Birmingham Activity Score and the Vasculitis Damage Index have become a core feature of clinical trials in ANCA-associated vasculitis (AAV) and formed the basis for sound clinical management of these complex conditions. We are still lacking accurate definitions of disease activity and damage progression in large-vessel vasculitis. There is an increasing interest in the role of patient-reported outcomes as a measure of disease impact; a disease-specific measure for use in AAV is being validated. We review how best to evaluate patients with large-, medium- and small-vessel vasculitis.


Inflammatory cytokines, angiogenesis, and fibrosis in the rat peritoneum.
Peritonitis, a common complication of peritoneal dialysis, is followed by acute changes in the function of the peritoneum. The role of inflammatory cytokines in these processes is not clearly identified. We used adenoviral-mediated gene transfer to transiently overexpress interleukin (IL)-1 beta (AdIL-1 beta) or tumor necrosis factor (TNF)-alpha (AdTNF-alpha) in the rat peritoneum then used a modified equilibrium test to study the histological and functional changes. Overexpression of IL-1 beta or TNF-alpha led to an acute inflammatory response. Both inflammatory cytokines induced an early expression of the angiogenic cytokine, vascular endothelial growth factor, along with increased expression of the profibrotic cytokine, transforming growth factor-beta1, along with fibronectin expression and collagen deposition in peritoneal tissues. Both inflammatory cytokines induced angiogenesis, increased solute permeability, and ultrafiltration dysfunction at earlier time points. Changes in structure and function seen in AdTNF-alpha-treated animals returned to normal by 21 days after infection, whereas AdIL-1 beta-treated animals had persistently increased vasculature with submesothelial thickening and fibrosis. This was associated with up-regulation TIMP-1. TNF-alpha or IL-1 beta both induce acute changes in the peritoneum that mimic those seen in peritoneal dialysis patients who experience an episode of peritonitis. These functional changes were associated with early angiogenesis that resolved rapidly after exposure to TNF-alpha. IL-1 beta exposure, however, led to a different response with sustained vascularization and fibrosis. IL-1 beta inhibition may be a therapeutic goal in acute peritonitis to prevent peritoneal damage.