Bone marrow mesenchymal stem cells do not enhance intra-synovial tendon healing despite engraftment and homing to niches within the synovium.
Intra-synovial tendon injuries display poor healing, which often results in reduced functionality and pain. A lack of effective therapeutic options has led to experimental approaches to augment natural tendon repair with autologous mesenchymal stem cells (MSCs) although the effects of the intra-synovial environment on the distribution, engraftment and functionality of implanted MSCs is not known. This study utilised a novel sheep model which, although in an anatomically different location, more accurately mimics the mechanical and synovial environment of the human rotator cuff, to determine the effects of intra-synovial implantation of MSCs. A lesion was made in the lateral border of the lateral branch of the ovine deep digital flexor tendon within the digital sheath and 2 weeks later 5 million autologous bone marrow MSCs were injected under ultrasound guidance into the digital sheath. Tendons were recovered post mortem at 1 day, and 1-2, 4, 12 and 24 weeks after MSC injection. For the 1-day and 1-2-week groups, MSCs labelled with fluorescent-conjugated magnetic iron-oxide nanoparticles (MIONs) were tracked with MRI, histology and flow cytometry. The 4, 12 and 24-week groups were implanted with non-labelled cells and compared with saline-injected controls for healing. The MSCs displayed no reduced viability in vitro to an uptake of 20.0 ± 4.6 pg MIONs per cell, which was detectable by MRI at minimal density of ~ 3 × 104 cells. Treated limbs indicated cellular distribution throughout the tendon synovial sheath but restricted to the synovial tissues, with no MSCs detected in the tendon or surgical lesion. The lesion was associated with negligible morbidity with minimal inflammation post surgery. Evaluation of both treated and control lesions showed no evidence of healing of the lesion at 4, 12 and 24 weeks on gross and histological examination. Unlike other laboratory animal models of tendon injury, this novel model mimics the failed tendon healing seen clinically intra-synovially. Importantly, however, implanted stem cells exhibited homing to synovium niches where they survived for at least 14 days. This phenomenon could be utilised in the development of novel physical or biological approaches to enhance localisation of cells in augmenting intra-synovial tendon repair.


Outpatient erbium:YAG (2940 nm) laser treatment for snoring: a prospective study on 40 patients.
Snoring is a sleep phenomenon due to the partial upper airway obstruction during sleep which causes vibration of the tissues of the rhino-oro-hypopharynx and less frequently the larynx. This study evaluated the use and effectiveness of the erbium:YAG 2940-nm laser as an adjunctive in providing treatment for patients suffering from chronic snoring-related sleep disorders. A prospective study of 40 consecutive patients with snoring and sleep disorders was performed, assessing data before and after three Er:YAG laser treatment sessions. During laser treatment, the pain was almost absent. There were no side effects, except a very mild sore throat in 1 out of 40 patients. The patient's evaluation of satisfaction of the results obtained after the treatments showed that 85% of cases were very satisfied, 5 patients (12.5%) reported being fairly satisfied with the treatment and only 1 subject (2.5%) was not satisfied. Mallampati, Friedman Tongue Position, and degree of O (oropharynx) at nose oropharynx hypopharynx and larynx classification were significantly decreased after the laser sessions. The decrease of Epworth Sleepiness Scale and Visual Analogue Scale for loudness of snoring, waking up during sleep because of snoring, dry mouth on waking, and choking was all statistically significant. The incidence of dreaming during the night also raised significantly; 30/40 (75%) of cases perceived less tightness in their throat and better breathing after treatment. These results were stable at 20 months follow-up (14-24 q) in 72% of cases. Nonsurgical and non-invasive Er:YAG laser treatment demonstrated to be a valid procedure in reducing the loudness of snoring.


Intelligent information loss: the coding of facial identity, head pose, and non-face information in the macaque face patch system.
Faces are a behaviorally important class of visual stimuli for primates. Recent work in macaque monkeys has identified six discrete face areas where most neurons have higher firing rates to images of faces compared with other objects (Tsao et al., 2006). While neurons in these areas appear to have different tuning (Freiwald and Tsao, 2010; Issa and DiCarlo, 2012), exactly what types of information and, consequently, which visual behaviors neural populations within each face area can support, is unknown. Here we use population decoding to better characterize three of these face patches (ML/MF, AL, and AM). We show that neural activity in all patches contains information that discriminates between the broad categories of face and nonface objects, individual faces, and nonface stimuli. Information is present in both high and lower firing rate regimes. However, there were significant differences between the patches, with the most anterior patch showing relatively weaker representation of nonface stimuli. Additionally, we find that pose-invariant face identity information increases as one moves to more anterior patches, while information about the orientation of the head decreases. Finally, we show that all the information we can extract from the population is present in patterns of activity across neurons, and there is relatively little information in the total activity of the population. These findings give new insight into the representations constructed by the face patch system and how they are successively transformed.


Acrosomal morphology as a novel criterion for male fertility diagnosis: relation with acrosin activity, morphology (strict criteria), and fertilization in vitro.
To determine the relationships between sperm acrosin activity, sperm morphology evaluated according to strict criteria, visually observed acrosomal morphology, and IVF rates. Prospective analytic study. Acrosin activity was determined on all semen samples together with a standard semen analysis. Emphasis was placed on sperm morphology and especially a novel criterion viz acrosome morphology (acrosome index) as recorded with bright field microscopy. University-based tertiary care center. Thirty-three couples undergoing IVF or GIFT with two or more metaphase II ova inseminated in vitro. In vitro fertilization rates of inseminated ova. Strong correlations were found between acrosome index, normal sperm morphology, and IVF rates. An acrosome index cutoff value could be established at > 10% normal acrosomes for IVF rates of > or = 50% (sensitivity and specificity = 100%) and an acrosin activity cutoff value at > 18 microIU/10(6) sperm. A multiple linear regression analysis showed that the acrosome index and acrosin activity added a significant contribution to the explanation of the variation in the fertilization rates. A strong positive correlation was found between acrosome index and IVF rates. Although the numbers of the study are small, the results indicate that the acrosome index possibly may be regarded as an additional tool in the prediction of IVF outcome and especially may be of value in the group of men with severe teratozoospermia, i.e., < or = 4% morphologically normal spermatozoa.


Auxin influx carriers control vascular patterning and xylem differentiation in Arabidopsis thaliana.
Auxin is an essential hormone for plant growth and development. Auxin influx carriers AUX1/LAX transport auxin into the cell, while auxin efflux carriers PIN pump it out of the cell. It is well established that efflux carriers play an important role in the shoot vascular patterning, yet the contribution of influx carriers to the shoot vasculature remains unknown. Here, we combined theoretical and experimental approaches to decipher the role of auxin influx carriers in the patterning and differentiation of vascular tissues in the Arabidopsis inflorescence stem. Our theoretical analysis predicts that influx carriers facilitate periodic patterning and modulate the periodicity of auxin maxima. In agreement, we observed fewer and more spaced vascular bundles in quadruple mutants plants of the auxin influx carriers aux1lax1lax2lax3. Furthermore, we show AUX1/LAX carriers promote xylem differentiation in both the shoot and the root tissues. Influx carriers increase cytoplasmic auxin signaling, and thereby differentiation. In addition to this cytoplasmic role of auxin, our computational simulations propose a role for extracellular auxin as an inhibitor of xylem differentiation. Altogether, our study shows that auxin influx carriers AUX1/LAX regulate vascular patterning and differentiation in plants.


Genome-informed Bradyrhizobium taxonomy: where to from here?
Bradyrhizobium is thought to be the largest and most diverse rhizobial genus, but this is not reflected in the number of described species. Although it was one of the first rhizobial genera recognised, its taxonomy remains complex. Various contemporary studies are showing that genome sequence information may simplify taxonomic decisions. Therefore, the growing availability of genomes for Bradyrhizobium will likely aid in the delineation and characterization of new species. In this study, we addressed two aims: first, we reviewed the availability and quality of available genomic resources for Bradyrhizobium. This was achieved by comparing genome sequences in terms of sequencing technologies used and estimated level of completeness for inclusion in genome-based phylogenetic analyses. Secondly, we utilized these genomes to investigate the taxonomic standing of Bradyrhizobium in light of its diverse lifestyles. Although genome sequences differed in terms of their quality and completeness, our data indicate that the use of these genome sequences is adequate for taxonomic purposes. By using these resources, we inferred a fully resolved, well-supported phylogeny. It separated Bradyrhizobium into seven lineages, three of which corresponded to the so-called supergroups known for the genus. Wide distribution of key lifestyle traits such as nodulation, nitrogen fixation and photosynthesis revealed that these traits have complicated evolutionary histories. We present the first robust Bradyrhizobium species phylogeny based on genome sequence information for investigating the evolution of this important assemblage of bacteria. Furthermore, this study provides the basis for using genome sequence information as a resource to make important taxonomic decisions, particularly at the species and genus levels.


Cyclin D1 stimulation of estrogen receptor transcriptional activity independent of cdk4.
Cyclin D1 plays an important role in the development of breast cancer and is required for normal breast cell proliferation and differentiation associated with pregnancy. We show that ectopic expression of cyclin D1 can stimulate the transcriptional activity of the estrogen receptor in the absence of estradiol and that this activity can be inhibited by 4-hydroxytamoxifen and ICI 182,780. Cyclin D1 can form a specific complex with the estrogen receptor. Stimulation of the estrogen receptor by cyclin D1 is independent of cyclin-dependent kinase 4 activation. Cyclin D1 may manifest its oncogenic potential in breast cancer in part through binding to the estrogen receptor and activation of the transcriptional activity of the receptor.


Tensor factorization for model-free space-variant blind deconvolution of the single- and multi-frame multi-spectral image.
The higher order orthogonal iteration (HOOI) is used for a single-frame and multi-frame space-variant blind deconvolution (BD) performed by factorization of the tensor of blurred multi-spectral image (MSI). This is achieved by conversion of BD into blind source separation (BSS), whereupon sources represent the original image and its spatial derivatives. The HOOI-based factorization enables an essentially unique solution of the related BSS problem with orthogonality constraints imposed on factors and the core tensor of the Tucker3 model of the image tensor. In contrast, the matrix factorization-based unique solution of the same BSS problem demands sources to be statistically independent or sparse which is not true. The consequence of such an approach to BD is that it virtually does not require a priori information about the possibly space-variant point spread function (PSF): neither its model nor size of its support. For the space-variant BD problem, MSI is divided into blocks whereupon the PSF is assumed to be a space-invariant within the blocks. The success of proposed concept is demonstrated in experimentally degraded images: defocused single-frame gray scale and red-green-blue (RGB) images, single-frame gray scale and RGB images blurred by atmospheric turbulence, and a single-frame RGB image blurred by a grating (photon sieve). A comparable or better performance is demonstrated in relation to the blind Richardson-Lucy algorithm which, however, requires a priori information about parametric model of the blur.


Induction of HT-29 Colon Cancer Cells Apoptosis by Pyrogallol with Growth Inhibiting Efficacy Against Drug-Resistant Helicobacter pylori.
Colon cancer is the most aggressive form of cancers, that causes 0.5 million deaths per year around the globe. Targeting colon cancer by conventional therapeutic options elicits toxicity. Traditional medicines take a lead to alleviate the existing clinical challenges. To investigate antibacterial activity against Helicobacter Pylori and in vitro anti-colon cancer activity by Acacia nilotica extract (ACE) and its active constituent pyrogallol. Pyrogallol isolated from A. nilotica by column chromatography and HPLC and structure was elucidated by spectral analysis. Antibacterial activity was done by flow cytometry. Cytotoxicity was measured by MTT assay. Apoptotic morphology and nuclear fragmentation were assessed with AO/ethidium bromide and DAPI staining. DNA fragmentation was done by electrophoresis. Western blot used to analyze the molecular mechanism of apoptosis. Cell cycle arrest was determined using flow cytometry of propidium iodide stained cells. Cell migration was determined by wound healing assay. ACE (20 µg/ml) and pyrogallol (10 µg/ml) treatment reduced the survival of H.pylori at 61% and 62%, respectively. MTT results show that HT-29 cells are more sensitive to pyrogallol with an IC50 value of 35μg/ml compared to ACE. Pyrogallol treated HT-29 cells reached dead state i.e. late apoptotic state with severe nuclear fragmentation. Pyrogallol elicits dose dependent DNA fragmentation in HT-29 cells. Pyrogallol induced apoptosis by simultaneous down-regulation of Bcl-2 and up-regulation of BAX and cytochrome c. Pyrogallol arrested HT-29 cells in S and G2/M phase of cell cycle. Further pyrogallol exhibited marked antimetastatic potential by inhibiting the migration of HT-29 cells dose dependently. Both ACE and pyrogallol repressed the growth of H.pylori and as significant anti-colon cancer agent.


Preventive and Therapeutic Vaccines against Human Papillomaviruses Associated Cervical Cancers.
Cervical cancer is, globally known to be, one of the most common cancers among women especially in developing countries. More than 90% of cervical cancers are associated with high-risk human papillomaviruses (HPVs) particularly HPV types 16 and 18. Two major strategies have been developed for prevention and treatment of cervical cancer and other HPV-associated malignancies; the first one is based on HPV virus-like particles (VLPs) containing HPV structural proteins. VLP based vaccines can induce genotype specific virus neutralizing antibodies for preventing HPV infections. The other strategy is based on HPV early genes especially E6 and E7 for eliminating the established HPV infections; therefore they are classified as HPV therapeutic vaccines. This article reviews the preventive and therapeutic vaccines against HPV infections and cervical cancer.


Surveillance of small aortic aneurysms does not alter anatomic suitability for endovascular repair.
Small abdominal aortic aneurysms (AAAs; 4-5.4 cm) are more likely to be suitable for endovascular aneurysm repair (EVAR) than large aortic aneurysms (>5.5 cm). The purpose of this study was to determine whether small AAA growth is associated with the development of morphologic characteristics that decrease eligibility for EVAR. We studied 54 patients who underwent 2 or more computed tomography scans with 3-dimensional reconstruction during surveillance of small AAAs. Morphologic aortic aneurysm features and changes were measured according to Society for Vascular Surgery reporting standards. Suitability for EVAR was determined by neck anatomy (diameter, length, and angulations), iliac artery morphology, and total aortic aneurysm angulation and tortuosity. The median age of the study cohort was 73 years (interquartile range [IQR], 65-77 years). The median follow-up period was 24 months (IQR, 15-36 months). The median small AAA diameter increased from 44.5 mm (IQR, 41-48 mm) to 48.9 mm (IQR, 45.7-52.0 mm). The median aortic neck diameter increased from 23.0 to 24.0 mm (P = .002), whereas median neck length decreased from 26.5 to 20.0 mm (P = .001). Aortic aneurysm median tortuosity index increased from 1.09 to 1.11 (P = .05). No significant changes in iliac artery morphology occurred. Overall, the anatomic suitability for endovascular repair did not significantly change during the study period (74% vs 69%; McNemar test; P = .25). Changes in aortic morphology are frequently associated with small AAA growth at mid-term follow-up, but such changes are minor and do not affect overall anatomic suitability for EVAR. These data reveal that continued surveillance of small AAAs does not threaten the window of opportunity for EVAR.


Bupropion perceived as a stimulant by two patients with a previous history of cocaine misuse.
Despite animal studies having shown a generalisation of the bupropion cue to cocaine, this drug has been used in cocaine abuse with mixed results. We here aimed at describing two cases which contradict current knowledge. We describe two cases of former cocaine abusers who reported a cocaine-like sensation upon taking bupropion. Bupropion improved patients' depression without any increase in cocaine craving. One of the patients increased without doctor consultation his dose on an as needed basis. The issue of bupropion cue generalisation to cocaine needs further elucidation. People with past cocaine addiction need to be informed on the potential of bupropion to elicit cocaine-like cues and be invited to adhere to medical prescription, because bupropion has been associated with fatalities in some cases.


Mothers' emotions after pediatric burn injury: Longitudinal associations with posttraumatic stress and depressive symptoms 18 months postburn.
Various emotions are implicated in posttraumatic stress disorder (PTSD). Longitudinal studies examining temporal associations between emotions and posttraumatic stress may reveal who is at risk of chronic psychological problems. This study examined the longitudinal relationships of mothers' trauma-related emotions with posttraumatic stress and depressive symptoms after pediatric burn injury. Data from two cohort studies were used (n = 296). Mothers reported the intensity of burn-related emotions within the first month (T1) and 12 months postburn (T2). The Impact of Event Scale (IES) and the Hospital and Anxiety Depression Scale (HADS-D; depression subscale) were administered at T1 and 18 months postburn (T3). Based on two exploratory factor analyses, emotion variables were combined into acute and long-term basic emotions (fear, sadness, horror, anger) and self-conscious emotions (guilt, shame). The path model showed a positive relationship between acute and long-term basic emotions. Higher long-term basic emotions were related to persistence of posttraumatic stress and depressive symptoms. Acute self-conscious emotions showed associations with posttraumatic stress and depressive symptoms at T1 and were longitudinally related to depressive, but not posttraumatic stress, symptoms. The posttraumatic stress measure was not based on DSM-5 PTSD criteria and results require replication using these criteria. This study suggests that mothers' acute self-conscious and long-term basic emotions in relation to their child's burn injury are involved in the development of posttraumatic stress and depressive symptoms. Clinically, assessing and monitoring parents' early posttraumatic stress, depressive symptoms and burn-related emotions may be useful to identify parents at risk.


Between empathy and anger: healthcare workers' perspectives on patient disengagement from antiretroviral treatment in Khayelitsha, South Africa - a qualitative study.
The benefits of long-term adherence to antiretroviral therapy (ART) are countered by interruptions in care or disengagement from care. Healthcare workers (HCWs) play an important role in patient engagement and negative or authoritarian attitudes can drive patients to disengage. However, little is known about HCWs' perspectives on disengagement. We explored HCWs' perspectives on ART disengagement in Khayelitsha, a peri-urban area in South Africa with a high HIV burden. Semi-structured interviews were conducted with 30 HCWs in a primary care HIV clinic to explore their perspectives of patients who disengage from ART. HCWs interviewed included clinical (doctors and nurses) and support staff (counsellors, social workers, data clerks, security guards, and occupational therapists). The interview guide asked HCWs about their experience working with patients who interrupt treatment and return to care. Transcripts were audio-recorded, transcribed, and analysed using an inductive thematic analysis approach. Most participants were knowledgeable about the complexities of disengagement and barriers to sustaining engagement with ART, raising their concerns that disengagement poses a significant public health problem. Participants expressed empathy for patients who interrupted treatment, particularly when the challenges that led to their disengagement were considered reasonable by the HCWs. However, many also expressed feelings of anger and frustration towards these patients, partly because they reported an increase in workload as a result. Some staff, mainly those taking chronic medication themselves, perceived patients who disengage from ART as not taking adequate responsibility for their own health. Lifelong engagement with HIV care is influenced by many factors including disclosure, family support, and HCW interactions. Findings from this study show that HCWs had contradictory feelings towards disengaged patients, experiencing both empathy and anger. Understanding this could contribute to the development of more nuanced interventions to support staff and encourage true person-centred care, to improve patient outcomes.


Herpes simplex viral vectors expressing Bcl-2 are neuroprotective when delivered after a stroke.
Considerable interest has focused on the possibility of using viral vectors to deliver genes to the central nervous system for the purpose of decreasing necrotic neuronal injury. To that end, we have previously shown that a herpes simplex virus (HSV) vector expressing Bcl-2 could protect neurons from ischemia. In that study, vector was delivered before the ischemia. However, for such gene therapy to be of clinical use, vectors must be protective even if delivered after the onset of the insult. In the present study, we show that an HSV vector expressing Bcl-2 protects striatal neurons when delivered after focal ischemia. Rats were exposed to middle cerebral artery occlusion for 1 hour, followed by reperfusion, and damage was assessed 48 hours later. Delivery of the Bcl-2 vector 30 minutes after reperfusion (i.e., 1.5 hours after ischemia onset) prevented any significant loss of virally-targeted neurons in the striatum. In contrast, in rats microinfused with a vector only expressing a reporter gene, a highly significant loss of neurons occurred. By 4 hours into the reperfusion period (5 hours after ischemia onset), delivery of the Bcl-2 vector was no longer protective. These data show the efficacy of postinsult gene therapy strategies for the brain, underline the finite length of this temporal therapeutic window, and support the growing evidence attesting to the neuroprotective potential of Bcl-2.


A Routine Activity Approach to Understand Cybergrooming Victimization Among Adolescents from Six Countries.
Little attention has been given academically to empirically tested theoretical frameworks that aim at measuring the risk of adolescents falling victim to cybergrooming. To this end, we have applied the routine activity theory (RAT) to investigate whether exposure to motivated offenders (PC/laptop ownership and Internet access in one's own bedroom), capable guardianship (parental mediation strategies of Internet use), and target suitability (adolescents' online disclosure of private information) might predict cybergrooming victimization among adolescents. Using data from a cross-sectional survey of 5,938 adolescents from Germany, India, South Korea, Spain, Thailand, and the United States, ranging in age from 12 to 18 (M = 14.77, SD = 1.60), we found that PC/laptop ownership and Internet access in one's own bedroom, parental mediation, and online disclosure are all directly associated with cybergrooming victimization. Although instructive parental mediation is negatively related to online disclosure and cybergrooming victimization, restrictive mediation is positively related to both. In addition, online disclosure partially mediated the relationship between parental mediation and cybergrooming victimization. The analyses confirm the effectiveness of applying RAT to cybergrooming. Moreover, this study highlights the need for prevention programs, including lessons on age-appropriate information and communication technology usage and access, to educate parents on using instructive strategies of Internet mediation, and inform adolescents about how to avoid disclosing too much private information online. RAT could function as a theoretical framework for these programs.


pH-Responsive nanocomposite fibres allowing MRI monitoring of drug release.
Magnetic resonance imaging (MRI) is one of the most widely-used non-invasive clinical imaging tools, producing detailed anatomical images whilst avoiding side effects such as trauma or X-ray radiation exposure. In this article, a new approach to non-invasive monitoring of drug release from a delivery vehicle via MRI was developed, using pH-responsive Eudragit L100 and S100 fibres encapsulating superparamagnetic iron oxide nanoparticles (SPIONs) and carmofur (a drug used in the treatment of colon cancer). Fibres were prepared by electrospinning, and found to be smooth and cylindrical with diameters of 645 ± 225 nm for L100 and 454 ± 133 nm for S100. The fibres exhibited pH responsive dissolution behaviour. Around the physiological pH range, clear pH-responsive proton relaxation rate changes due to matrix swelling/dissolution can be observed: r2 values of L100 fibres increase from 29.3 ± 8.3 to 69.8 ± 2.5 mM-1 s-1 over 3 h immersion in a pH 7.4 medium, and from 13.5 ± 2.0 mM-1 s-1 to 42.1 ± 3.0 mM-1 s-1 at pH 6.5. The r2 values of S100 fibres grow from 30.4 ± 4.4 to 64.7 ± 1.0 mM-1 s-1 at pH 7.4, but at pH 6.5, where the S100 fibres are not soluble, r2 remains very low (< 4 mM-1 s-1). These dramatic changes in relaxivity demonstrate that pH-responsive dissolution results in SPION release. In vitro drug release studies showed the formulations gave rapid release of carmofur at physiological pH values (pH 6.5 and 7.4), and acid stability studies revealed that they can protect the SPIONs from digestion in acid environments, giving the fibres potential for oral administration. Exploration of the relationship between relaxivity and carmofur release suggests a linear correlation (R2 > 0.94) between the two. Mathematical equations were developed to predict carmofur release in vitro, with very similar experimental and predicted release profiles obtained. Therefore, the formulations developed herein have the potential to be used for non-invasive monitoring of drug release in vivo, and could ultimately result in dramatic reductions to off-target side effects from interventions such as chemotherapy.


Preparing dental graduates to provide care to individuals with special needs.
In 2004, the Commission on Dental Accreditation (CODA) adopted a new standard that directs dental and dental hygiene programs to prepare dental professionals for the care of persons with special health care needs. This article reviews the demographics of individuals with special needs, documents that most dental schools provide their students with very limited educational opportunities related to the care of this population, describes the path that was followed to bring about change in the accrediting standard, and discusses the difficulties involved in developing the needed educational programs. Educational programs at two dental schools are presented as examples of how schools can provide students with learning experiences pertinent to the new CODA standard that states: "Graduates must be competent in assessing the treatment needs of patients with special needs."


Select nutrients in the ovine uterine lumen. IV. Expression of neutral and acidic amino acid transporters in ovine uteri and peri-implantation conceptuses.
The availability of specific neutral and acidic amino acids in the uterine lumen of ewes increased significantly during the peri-implantation period, but mechanisms for their transport into the uterine lumen and uptake by conceptuses are not established in any species. In this study, effects of pregnancy, progesterone (P4), and interferon tau (IFNT) on expression of neutral and acidic amino acid transporters in uteri of cyclic and pregnant ewes and conceptuses were studied. SLC1A2, SLC1A3, SLC3A1, SLC6A14, SLC6A19, SLC7A6, SLC38A3, and SLC38A6 mRNAs were only weakly expressed in the ovine endometrium. However, SLC1A4, SLC1A5, SLC7A8, and SLC43A2 mRNAs were detectable in uterine luminal epithelia (LE), superficial glandular epithelia (sGE), and/or glandular epithelia (GE). SLC1A1 and SLC7A5 mRNAs were most abundant in LE/sGE and GE. SLC1A3 and SLC38A4 mRNAs were most abundant in uterine stroma. SLC38A6 mRNA was detected only in cells with a stromal distribution suggesting immune lineage. SLC1A5 mRNA was expressed primarily in LE/sGE and stromal cells, and it was more abundant in uteri of pregnant ewes (day x status interaction; P < 0.05). Furthermore, P4 induced and IFNT further stimulated SLC1A5 expression in LE/sGE. Endometrial SLC1A1, SLC7A5, and SLC43A2 mRNAs demonstrated both temporal and cellSLC-specific changes. Several mRNAs were detectable in trophectoderm (SLC6A19, SLC7A5, SLC7A6, and SLC43A2), while others were more abundant in endoderm (SLC1A4, SLC1A5, SLC6A19, SLC7A5, SLC7A6, SLC7A8, and SLC43A2) of conceptuses. These results document coordinate changes in expression of transporters that are likely responsible for increases in amounts of neutral and acidic amino acids in the uterine lumen to support conceptus growth, development, and survival.


Acute traumatic compartment syndrome of the leg in children: diagnosis and outcome.
Currently, the most common clinical scenario for compartment syndrome in children is acute traumatic compartment syndrome of the leg. We studied the cause, diagnosis, treatment, and outcome of acute traumatic compartment syndrome of the leg in children. Forty-three cases of acute traumatic compartment syndrome of the leg in forty-two skeletally immature patients were collected from two large pediatric trauma centers over a seventeen-year period. All children with acute traumatic compartment syndrome underwent fasciotomy. The mechanism of injury, date and time of injury, time to diagnosis, compartment pressures, time to fasciotomy, and outcome at the time of the latest follow-up were recorded. Thirty-five (83%) of the forty-two patients were injured in a motor-vehicle accident and sustained tibial and fibular fractures. The average time from injury to fasciotomy was 20.5 hours (range, 3.9 to 118 hours). In general, the functional outcome was excellent at the time of the latest follow-up. No cases of infection were noted when fasciotomy was performed long after the injury. At the time of the latest follow-up, forty-one (95%) of forty-three cases were associated with no sequelae (such as pain, loss of function, or decreased sensation). The two patients who lost function had fasciotomy 82.5 and eighty-six hours after the injury. Despite a long period from injury to fasciotomy, most children who are managed for acute traumatic compartment syndrome of the leg have an excellent outcome. This delay may occur because acute traumatic compartment syndrome manifests itself more slowly in children or because the diagnosis is harder to establish in this age group. The results of the present study should raise awareness of late presentation and the importance of vigilance for developing compartment syndrome in the early days after injury. Fasciotomy during the acute swelling phase, even long after injury, produced excellent results with no cases of infection.


The two tryptophans of β2-microglobulin have distinct roles in function and folding and might represent two independent responses to evolutionary pressure.
We have recently discovered that the two tryptophans of human β2-microglobulin have distinctive roles within the structure and function of the protein. Deeply buried in the core, Trp95 is essential for folding stability, whereas Trp60, which is solvent-exposed, plays a crucial role in promoting the binding of β2-microglobulin to the heavy chain of the class I major histocompatibility complex (MHCI). We have previously shown that the thermodynamic disadvantage of having Trp60 exposed on the surface is counter-balanced by the perfect fit between it and a cavity within the MHCI heavy chain that contributes significantly to the functional stabilization of the MHCI. Therefore, based on the peculiar differences of the two tryptophans, we have analysed the evolution of β2-microglobulin with respect to these residues. Having defined the β2-microglobulin protein family, we performed multiple sequence alignments and analysed the residue conservation in homologous proteins to generate a phylogenetic tree. Our results indicate that Trp60 is highly conserved, whereas some species have a Leu in position 95; the replacement of Trp95 with Leu destabilizes β2-microglobulin by 1 kcal/mol and accelerates the kinetics of unfolding. Both thermodynamic and kinetic data fit with the crystallographic structure of the Trp95Leu variant, which shows how the hydrophobic cavity of the wild-type protein is completely occupied by Trp95, but is only half filled by Leu95. We have established that the functional Trp60 has been present within the sequence of β2-microglobulin since the evolutionary appearance of proteins responsible for acquired immunity, whereas the structural Trp95 was selected and stabilized, most likely, for its capacity to fully occupy an internal cavity of the protein thereby creating a better stabilization of its folded state.


Effects of intracerebroventricular injection of histamine on memory deficits induced by hippocampal lesions in rats.
The influence of bilateral hippocampal lesions on active avoidance response was studied in rats, as well as the effect of intracerebroventricular (i.c.v.) injection of histamine on memory deficits caused by hippocampectomy. Retardation of learning acquisition was produced by lesioning of the bilateral dorsal hippocampus in active avoidance response. Memory retention was also impaired by hippocampectomy. Although locomotor activity and rearing behavior measured by open-field test increased after hippocampal lesions, there was no relation between impairment of learning and increase in exploratory behavior. I.c.v. injection of histamine and i.p. injection of histidine resulted in an improvement of memory deficits (not only learning acquisition but also memory retrieval) induced by hippocampal lesions in rats. Histamine contents of the hippocampus and hypothalamus decreased after hippocampectomy, and a decrease in histamine contents of both areas was restored by histamine (i.c.v.) and histidine (i.p.) injection. In addition, a close relationship was found between decrease in response latency of avoidance response and an increase in histamine content of the hippocampus and hypothalamus after histamine injection.


Evaluating youth-friendly health services: young people's perspectives from a simulated client study in urban South Africa.
Few§These authors contributed equally to this work. youth-friendly health services worldwide have been scaled up or evaluated from young people's perspectives. South Africa's Youth Friendly Services (YFS) programme is one of the few to have been scaled up. This study investigated young people's experiences of using sexual and reproductive health services at clinics providing the YFS programme, compared to those that did not, using the simulated client method. Fifteen primary healthcare clinics in Soweto were randomly sampled: seven provided the YFS programme. Simulated clients conducted 58 visits; young men requested information on condom reliability and young women on contraceptive methods. There were two outcome measures: a single measure of the overall clinic experience (clinic visit score) and whether or not simulated clients would recommend a clinic to their peers. The clinic visit score was based on variables relating to the simulated clients' interactions with staff, details of their consultation, privacy, confidentiality, the healthcare workers' characteristics, and the clinic environment. A larger score corresponds to a worse experience than a smaller one. Multilevel regression models and framework analysis were used to investigate young people's experiences. Health facilities providing the YFS programme did not deliver a more positive experience to young people than those not providing the programme (mean difference in clinic visit score: -0.18, 95% CI: -0.95, 0.60, p=0.656). They were also no more likely to be recommended by simulated clients to their peers (odds ratio: 0.48, 95% CI: 0.11, 2.10, p=0.331). More positive experiences were characterised by young people as those where healthcare workers were friendly, respectful, knew how to talk to young people, and appeared to value them seeking health information. Less positive experiences were characterised by having to show soiled sanitary products to obtain contraceptives, healthcare workers expressing negative opinions about young people seeking information, lack of privacy, and inadequate information. The provision and impact of the YFS programme are limited. Future research should explore implementation. Regular training and monitoring could enable healthcare workers to address young people's needs.


Standing genetic variation fuels rapid adaptation to ocean acidification.
Global climate change has intensified the need to assess the capacity for natural populations to adapt to abrupt shifts in the environment. Reductions in seawater pH constitute a conspicuous global change stressor that is affecting marine ecosystems globally. Here, we quantify the phenotypic and genetic modifications associated with rapid adaptation to reduced seawater pH in the Mediterranean mussel, Mytilus galloprovincialis. We reared a genetically diverse larval population in two pH treatments (pHT 8.1 and 7.4) and tracked changes in the shell-size distribution and genetic variation through settlement. Additionally, we identified differences in the signatures of selection on shell growth in each pH environment. Both phenotypic and genetic data show that standing variation can facilitate adaptation to declines in seawater pH. This work provides insight into the processes underpinning rapid evolution, and demonstrates the importance of maintaining variation within natural populations to bolster species' adaptive capacity as global change progresses.


MRI of the brain with ultra-short echo-time pulse sequences.
As well as the long-T2 relaxation components normally detected with conventional imaging techniques, the brain has short-T2 components. We wished to use ultra-short (0.08 ms) echo time (UTE) pulse sequences to assess the feasibility of imaging these in normal subjects and patients. UTE sequences were employed, with or without fat suppression, 90 degree long-T2 suppression pulses, and selective nulling of long-T2 components using an inversion pulse. Subtraction of later echoes from the first was also used to reduce the signal from long-T2 components. We studied dive normal subjects and 15 patients with various diseases. Short-T2 components were demonstrated in grey and white matter. Increased signal from these components was seen in meningeal disease, probable calcification, presumed cavernomas, melanoma metastases and probable gliosis. Reduced signal was seen in some tumours, infarcts, mild multifocal vascular disease and vasogenic oedema. Further development and evaluation of these pulse sequences is warranted.


Enhancer of zeste homolog 2 (EZH2) expression is an independent prognostic factor in renal cell carcinoma.
The enhancer of zeste homolog 2 (EZH2) gene exerts oncogene-like activities and its (over)expression has been linked to several human malignancies. Here, we studied a possible association between EZH2 expression and prognosis in patients with renal cell carcinoma (RCC). EZH2 protein expression in RCC specimens was analyzed by immunohistochemistry using a tissue microarray (TMA) containing RCC tumor tissue and corresponding normal tissue samples of 520 patients. For immunohistochemical assessment of EZH2 expression, nuclear staining quantity was evaluated using a semiquantitative score. The effect of EZH2 expression on cancer specific survival (CSS) was assessed by univariate and multivariate Cox regression analyses. During follow-up, 147 patients (28%) had died of their disease, median follow-up of patients still alive was 6.0 years (range 0-16.1 years). EZH2 nuclear staining was present in tumor cores of 411 (79%) patients. A multivariate Cox regression analysis revealed that high nuclear EZH2 expression was an independent predictor of poor CSS (> 25-50% vs. 0%: HR 2.72, p = 0.025) in patients suffering from non-metastatic RCC. Apart from high nuclear EZH2 expression, tumor stage and Fuhrman's grading emerged as significant prognostic markers. In metastatic disease, nuclear EZH2 expression and histopathological subtype were independent predictive parameters of poor CSS (EZH2: 1-5%: HR 2.63, p = 0.043, >5-25%: HR 3.35, p = 0.013, >25%-50%: HR 4.92, p = 0.003, all compared to 0%: HR 0.36, p = 0.025, respectively). This study defines EZH2 as a powerful independent unfavourable prognostic marker of CSS in patients with metastatic and non-metastatic RCC.


Surgical Decision Making for Acetabular Labral Tears: An International Perspective.
Studies comparing acetabular labral repair with debridement have generally demonstrated better clinical outcomes with repair. However, it is not clear whether hip arthroscopic surgeons agree on the indications for labral repair and debridement, bringing the generalizability of these (and future) studies into question. To investigate surgical decision making for acetabular labral tears, with a specific focus on indications for repair or debridement, by performing an international survey of hip arthroscopic surgeons. Cross-sectional study. A total of 35 hip arthroscopic surgeons from around the world were invited to participate in this survey study. Surgeons selected the factors that they consider when deciding to repair or debride acetabular labral tears. For each variable selected, computerized adaptive logic prompted additional questions to better define how that variable affects decision making. Six deidentified intraoperative videos of a variety of labral tears were included to determine the level of agreement between the experts on which labral tears are repairable. The survey response rate was 86%. A majority (77%) of hip arthroscopic surgeons selected the intraoperative appearance of the labrum as the most important factor affecting their decision making. Specific factors affecting surgical decision making included pattern of the labral tear (73% of surgeons), ossification of the labrum (70%), magnetic resonance imaging findings (70%), patient age (63%), activity level (57%), radiographic findings (53%), calcification of the labrum (50%), and thickness of the labrum (47%). Three intraoperative videos had ≥90% agreement for labral repair, while the other 3 had ≤76% agreement. The intraoperative appearance of the labrum is the most important factor affecting surgical decision making. However, different surgeons viewing the same tear arthroscopically may select different treatments. The indications to repair a torn acetabular labrum are highly variable among hip arthroscopic surgeons.


Elongation and preload stress in dental implant abutment screws.
A common problem associated with dental implant restorations is loosening of screws that retain the prosthesis to the implant. A method was developed to determine initial preload on UCLA-type abutment screws by measuring elongation after applying known tightening torques with a digital torque gauge. Loosening torque was also measured after tightening to 32 N-cm torque for gold alloy abutment screws and 20 N-cm for titanium abutment screws. Gold alloy and titanium abutment screws were each used to secure a gold UCLA hexed abutment to a titanium implant. Stresses and forces were calculated from the elongation measurements for three regions of each screw. Elongation of the screws after applying the manufacturer's recommended tightening torques were within the elastic range. Induced stresses were 57.5% and 56% of the yield strengths for gold alloy and titanium, respectively. Tightening of screws beyond recommended levels may be possible without producing plastic deformation. At manufacturer's recommended torques, mean preload was 468.2 (+/- 57.9) N using gold alloy screws and 381.5 (+/- 72.9) N with titanium screws.


Narrative Tensions in Strained Junior Elite Performers' Experiences of Becoming Elite Performers.
Contextualized within narrative theory and the field of talent identification and development systems (TIDS), this interview study examined strained junior elite performers' experiences of becoming elite performers while participating in prestigious national TIDS. The study explored how junior elite performers perceive and negotiate their personal narratives of becoming within a cultural master narrative of being. The focus is on how the quality of person-environment interaction, characterized by narrative alignment or tensions, relates to perceptions of identity, agency, and physical and mental health. We purposefully recruited eight participants (Mage = 17.31, SD = 0.9) from a previously published study, who reported experiencing suboptimal psychological functioning compared with their peers to explore narrative tensions in their storylines. The data were collected through semi-structured interviews and examined, using narrative analysis. We identified "the performance narrative" as the dominating cultural narrative within the TIDS and three distinct personal narratives of negotiation with unique characteristics: obsessive and externally driven alignment - "striving to stay at the top of the game"; tensions - "just hanging in there"; and disruption from alignment - "when the going gets tough." The results indicated that tensions and lack of alignment between the dominating cultural narrative and the individual narrative seem to increase the risk of experiencing identity challenges, suboptimal functioning, and aspects of ill-being. The study offers critical reflections on the dominating performance narrative within TIDS and additionally suggests an alternative athlete-centered and more holistic approach that combines both personal and performance developments.


Reporting disease control rates or clinical benefit rates in early clinical trials of anticancer agents: useful endpoint or hype?
Disease Control Rate (DCR) and Clinical Benefit Rate (CBR) are defined as the percentage of patients with advanced or metastatic cancer who have achieved complete response, partial response and stable disease to a therapeutic intervention in clinical trials of anticancer agents. DCR and CBR are commonly reported in many clinical trials in abstracts, papers, meeting presentations and media releases. The frequent use of these measures of drug activity presents the question of whether DCR and CBR are useful additional endpoints in early clinical trials, and if they can reasonably predict the success of an agent in subsequent, adequately powered, randomized trials. There are no comprehensive analyses to demonstrate that CBR and DCR add to the value of traditional response/activity endpoints in early clinical trials. Data from phase II clinical trials in which the DCR or CBR are reported suggest that DCR or CBR provides ambiguous information that likely exaggerates the anticancer activity of the therapy. The terms 'disease control' and 'clinical benefit' in the context of non-randomized trials are themselves disingenuous because neither tumor regression nor stable disease, defined without any consideration of duration of effect or reduction of symptoms appropriate for the specific patient population, are evidence of these endpoints in an individual patient.


Cathepsin B Cysteine Proteinase is Essential for the Development and Pathogenesis of the Plant Parasitic Nematode Radopholus similis.
Radopholus similis is an important plant parasitic nematode which severely harms many crops. Cathepsin B is present in a wide variety of organisms, and plays an important role in many parasites. Understanding cathepsin B of R. similis would allow us to find new targets and approaches for its control. In this study, we found that Rs-cb-1 mRNA was expressed in esophageal glands, intestines and gonads of females, testes of males, juveniles and eggs in R. similis. Rs-cb-1 expression was the highest in females, followed by juveniles and eggs, and was the lowest in males. The maximal enzyme activity of Rs-CB-1 was detected at pH 6.0 and 40 °C. Silencing of Rs-cb-1 using in vitro RNAi (Soaking with dsRNA in vitro) not only significantly inhibited the development and hatching of R. similis, but also greatly reduced its pathogenicity. Using in planta RNAi, we confirmed that Rs-cb-1 expression in nematodes were significantly suppressed and the resistance to R. similis was significantly improved in T2 generation transgenic tobacco plants expressing Rs-cb-1 dsRNA. The genetic effects of in planta RNAi-induced gene silencing could be maintained in the absence of dsRNA for at least two generations before being lost, which was not the case for the effects induced by in vitro RNAi. Overall, our results first indicate that Rs-cb-1 plays key roles in the development, hatching and pathogenesis of R. similis, and that in planta RNAi is an effective tool in studying gene function and genetic engineering of plant resistance to migratory plant parasitic nematodes.


Structured dietary advice incorporating walnuts achieves optimal fat and energy balance in patients with type 2 diabetes mellitus.
A cardioprotective dietary fat profile is recommended for the treatment of type 2 diabetes. The clinical feasibility of advice strategies targeting specific fatty acid intakes and the extent to which they can be achieved by free-living populations needs to be tested. Walnuts, with high n-3 polyunsaturated fatty acid (PUFA) content, may help optimize fatty acid intakes, but regular consumption might increase total fat and energy intakes. This study examined whether advice that refers to a total dietary pattern inclusive of walnuts would result in low-fat energy-controlled diets with optimal dietary fat proportions for patients with type 2 diabetes mellitus. A parallel-design, controlled trial was completed by 55 free-living men and women with established type 2 diabetes mellitus. Participants were randomly assigned to one of three groups: low-fat (general advice), modified low-fat (total diet advice using exchange lists to differentiate PUFA-rich foods), walnut-specific (modified low fat including 30 g walnuts/day). Dietary intakes and clinical outcomes were measured at baseline, and at 3 and 6 months. Dietary goals were: less than 10% of energy from saturated fat, 7% to 10% of energy from PUFA, adequate n-3 PUFA (>or=2.22 g alpha-linolenic acid, >or=0.65 g eicosapentaenoic acid [EPA]+docosahexaenoic acid [DHA]) and n-6 to n-3 ratio less than 10. The proportion of subjects achieving dietary goals and major food sources of fat were determined. At baseline, dietary intakes were not significantly different between groups. No group and few individuals (10%) were consuming adequate PUFA, with meat the main source of dietary fat (22% total dietary fat). At 3 and 6 months, energy and macronutrient intakes were similar among groups. The walnut group, however, was the only group to achieve all fatty acid intake targets (P <.01), and had the greatest proportion of subjects achieving targets ( P <.05). Walnuts were the main source of dietary fat (31%) and n-3 PUFA (50%), while 350 g oily fish/day provided a further 17% n-3 PUFA consumed by this group. Specific advice for the regular inclusion of walnuts in the context of the total diet helps achieve optimal fat intake proportions without adverse effects on total fat or energy intakes in patients with type 2 diabetes mellitus.


Implications of Von Hippel-Lindau Syndrome and Renal Cell Carcinoma.
Von Hippel-Lindau syndrome (VHLS) is a rare hereditary neoplastic disorder caused by mutations in the vhl gene leading to the development of tumors in several organs including the central nervous system, pancreas, kidneys, and reproductive organs. Manifestations of VHLS can present at different ages based on the affected organ and subclass of disease. In the subclasses of VHLS that cause renal disease, renal involvement typically begins closer to the end of the second decade of life and can present in different ways ranging from simple cystic lesions to solid tumors. Mutations in vhl are most often associated with clear cell renal carcinoma, the most common type of renal cancer, and also play a major role in sporadic cases of clear cell renal carcinoma. The recurrent, multifocal nature of this disease presents difficult challenges in the long-term management of patients with VHLS. Optimization of renal function warrants the use of several different approaches common to the management of renal carcinoma such as nephron sparing surgery, enucleation, ablation, and targeted therapies. In VHLS, renal lesions of 3 cm or bigger are considered to have metastatic potential and even small lesions often harbor malignancy. Many of the aspects of management revolve around optimizing both oncologic outcome and long-term renal function. As new surgical strategies and targeted therapies develop, the management of this complex disease evolves. This review will discuss the key aspects of the current management of VHLS.


Systematic review of treatments for psoriatic arthritis: 2014 update for the GRAPPA.
Psoriatic arthritis (PsA) is a chronic systemic inflammatory disorder characterized by the association of arthritis and periarticular inflammation in patients with psoriasis. In addition to a heterogeneous and variable clinical course, PsA is complex and multifaceted and may include prominent involvement in the peripheral and axial diarthrodial joints, the skin and nails, and in periarticular structures such as entheses. Simultaneous inflammation in the skin and musculoskeletal structures in a single patient, a relatively common scenario, often leads to marked decrease in function and quality of life. Thus, it is essential for the clinician to document the extent of disease involvement and craft a therapeutic plan that addresses the different domains of disease. In an effort to update previous treatment recommendations developed by the Group for Research and Assessment of Psoriasis and Psoriatic Arthritis (GRAPPA), several evidence-based, systemic reviews of therapies for PsA were completed, analyzed, and circulated for consensus.


A comparison of distribution curves of body mass index from Brazil and the United States for assessing overweight and obesity in Brazilian adolescents.
To assess the validity of recommendations for use of the 85th and 95th percentiles of body mass index (BMI) of the population in the United States of America as a screening tool to assess overweight/obesity in adolescents. We investigated the relation between BMI and percent body fat in 1,540 adolescents (717 males and 823 females) aged 10 to 17.9 years old from a private high school in Niterói, a city in the state of Rio de Janeiro, Brazil. We used bioelectric impedance, with the appropriate equations for adolescents, to estimate percent body fat, which served as the gold standard (30% for girls and 25% for boys) to calculate the sensitivity and specificity of the 85th and 95th percentiles of the United States and Brazilian distribution curves of BMI. Sensitivity and specificity were high (above 80%) for the Niterói boys, except for the 85th percentile of the Brazilian curve (specificity = 61.8%) and for the 95th percentile of the United States curve (sensitivity = 55.4%). For the Niterói girls, the 85th- and 95th-percentile BMI cutoff points, from both the United States and Brazilian curves, showed low sensitivity, and that sensitivity decreased with age. Specificity was high for the girls, and much higher than it was for the boys. These data suggest that using BMI to screen for overweight/obesity in adolescents can generate a high percentage of false-positives for Niterói boys and an even higher percentage of false-negatives for Niterói girls. A more universal approach to using anthropometric measures to screen for overweight/obesity should be developed, preferably linked to stages of maturation.


Chiral Redox-Active Isosceles Triangles.
Designing small-molecule organic redox-active materials, with potential applications in energy storage, has received considerable interest of late. Herein, we report on the synthesis, characterization, and application of two rigid chiral triangles, each of which consist of non-identical pyromellitic diimide (PMDI) and naphthalene diimide (NDI)-based redox-active units. (1)H and (13)C NMR spectroscopic investigations in solution confirm the lower symmetry (C2 point group) associated with these two isosceles triangles. Single-crystal X-ray diffraction analyses reveal their rigid triangular prism-like geometries. Unlike previously investigated equilateral triangle containing three identical NDI subunits, both isosceles triangles do not choose to form one-dimensional supramolecular nanotubes by dint of [C-H···O] interaction-driven columnar stacking. The rigid isosceles triangle, composed of one NDI and two PMDI subunits, forms-in the presence of N,N-dimethylformamide-two different types of intermolecular NDI-NDI and NDI-PMDI π-π stacked dimers with opposite helicities in the solid state. Cyclic voltammetry reveals that both isosceles triangles can accept reversibly up to six electrons. Continuous-wave electron paramagnetic resonance and electron-nuclear double-resonance spectroscopic investigations, supported by density functional theory calculations, on the single-electron reduced radical anions of the isosceles triangles confirm the selective sharing of unpaired electrons among adjacent redox-active NDI subunit(s) within both molecules. The isosceles triangles have been employed as electrode-active materials in organic rechargeable lithium-ion batteries. The evaluation of the structure-performance relationships of this series of diimide-based triangles reveals that the increase in the number of NDI subunits, replacing PMDI ones, within the molecules improves the electrochemical cell performance of the batteries.


Impaired microRNA processing causes corpus luteum insufficiency and infertility in mice.
The microRNA (miRNA) processing enzyme Dicer1 is required for zygotic and embryonic development, but the early embryonic lethality of Dicer1 null alleles in mice has limited our ability to address the role of Dicer1 in normal mouse growth and development. To address this question, we used a mouse mutant with a hypomorphic Dicer1 allele (Dicer(d/d)) and found that Dicer1 deficiency resulted in female infertility. This defect in female Dicer(d/d) mice was caused by corpus luteum (CL) insufficiency and resulted, at least in part, from the impaired growth of new capillary vessels in the ovary. We found that the impaired CL angiogenesis in Dicer(d/d) mice was associated with a lack of miR17-5p and let7b, 2 miRNAs that participate in angiogenesis by regulating the expression of the antiangiogenic factor tissue inhibitor of metalloproteinase 1. Furthermore, injection of miR17-5p and let7b into the ovaries of Dicer(d/d) mice partially normalized tissue inhibitor of metalloproteinase 1 expression and CL angiogenesis. Our data indicate that the development and function of the ovarian CL is a physiological process that appears to be regulated by miRNAs and requires Dicer1 function.


Race- and gender-related differences in clinical characteristics and quality of life among outpatients with psychotic disorders.
Historically, the literature suggests that African Americans with mental illness are diagnosed with psychotic disorders at a higher rate and receive higher doses of antipsychotic medications than other racial groups. However, few studies have compared clinical characteristics and quality of life among African-American (AA) and white men and women. Thus, research is needed to examine potential race and gender differences in clinical characteristics, prescribing practices, and quality of life. This exploratory, hypothesis-generating study examined current and past diagnoses, current pharmacotherapy, failed psychotropic medications, and quality of life among 23 AA and 31 white men and women receiving outpatient treatment for psychosis. Depression and psychotic depression were common complaints in the sample, yet only a third of the patients received antidepressants. We found that AA men received an antidepressant for depression symptoms less often, received higher antipsychotic doses, and rated their overall quality of life as poorer than any other group. White men and AA women had a history of more years of mental illness and had experienced 57% and 69% more psychotropic medication failures, respectively, than AA men or white women. Quality of life scores were significantly related to years of mental illness, number of past diagnoses, and number of failed medications. Our data suggest that clinicians could significantly enhance prognostic outcomes in outpatients with psychotic disorders by routinely re-evaluating depressive symptomatology and prescribing practices and considering adding psychosocial interventions to avert deterioration in quality of life. Further investigation of race and gender differences in quality of life and satisfaction as a function of diagnoses and treatment is warranted.


Discovering local patterns of co-evolution: computational aspects and biological examples.
Co-evolution is the process in which two (or more) sets of orthologs exhibit a similar or correlative pattern of evolution. Co-evolution is a powerful way to learn about the functional interdependencies between sets of genes and cellular functions and to predict physical interactions. More generally, it can be used for answering fundamental questions about the evolution of biological systems.Orthologs that exhibit a strong signal of co-evolution in a certain part of the evolutionary tree may show a mild signal of co-evolution in other branches of the tree. The major reasons for this phenomenon are noise in the biological input, genes that gain or lose functions, and the fact that some measures of co-evolution relate to rare events such as positive selection. Previous publications in the field dealt with the problem of finding sets of genes that co-evolved along an entire underlying phylogenetic tree, without considering the fact that often co-evolution is local. In this work, we describe a new set of biological problems that are related to finding patterns of local co-evolution. We discuss their computational complexity and design algorithms for solving them. These algorithms outperform other bi-clustering methods as they are designed specifically for solving the set of problems mentioned above.We use our approach to trace the co-evolution of fungal, eukaryotic, and mammalian genes at high resolution across the different parts of the corresponding phylogenetic trees. Specifically, we discover regions in the fungi tree that are enriched with positive evolution. We show that metabolic genes exhibit a remarkable level of co-evolution and different patterns of co-evolution in various biological datasets.In addition, we find that protein complexes that are related to gene expression exhibit non-homogenous levels of co-evolution across different parts of the fungi evolutionary line. In the case of mammalian evolution, signaling pathways that are related to neurotransmission exhibit a relatively higher level of co-evolution along the primate subtree. We show that finding local patterns of co-evolution is a computationally challenging task and we offer novel algorithms that allow us to solve this problem, thus opening a new approach for analyzing the evolution of biological systems.


Increased Global DNA Methylation and Decreased TGFβ1 Promoter Methylation in Glaucomatous Lamina Cribrosa Cells.
Glaucoma is an optic neuropathy that affects 60 million people worldwide. There is an underlying fibrosis associated with the lamina cribrosa (LC) in glaucoma. DNA methylation is well established in regulating fibrosis and may be a therapeutic target for glaucoma. The purpose of this study was to compare global DNA methylation levels in primary human normal (NLC) and glaucomatous (GLC) cells, and to investigate DNA methylation in driving fibrosis through regulation of transforming growth factor β1 (TGFβ1). LC cells were cultured from normal and glaucomatous human donors. Global methylation was assessed by ELISA. qPCR was conducted for DNA methyltransferases (DNMTs), methyl-CpG-binding protein 2 (MeCP2), TGFβ 1 and 2, collagen 1α1 (COL1A1), and α-smooth muscle actin (αSMA). TGFβ1 and DNMT1 were examined by immunofluorescence. Methylation of the TGFβ1 promoter was determined by methylation-specific PCR (MSP). Global DNA methylation demonstrated an increase in GLC compared with NLC cells (P<0.05). The previously mentioned methylation and matrix genes were increased in GLC compared with NLC cells (P<0.05). Immunofluorescence showed increased TGFβ1 and DNMT1 in GLC compared with NLC cells. MSP showed increased unmethylated DNA in the TGFβ1 promoter of GLC compared with NLC cells. We found increased expression of fibrotic genes in GLC cells and demonstrated an increase in global DNA methylation and in associated enzymes in GLC cells. Furthermore, we showed decreased promoter methylation of TGFβ1 in GLC cells. Determining a role for methylation in glaucoma and in regulating TGFβ1 may provide a novel therapeutic approach.


Karyological characterization of the endemic Iberian rock lizard, Iberolacerta monticola (Squamata, Lacertidae): insights into sex chromosome evolution.
Rock lizards of the genus Iberolacerta constitute a promising model to examine the process of sex chromosome evolution, as these closely related taxa exhibit remarkable diversity in the degree of sex chromosome differentiation with no clear phylogenetic segregation, ranging from cryptic to highly heteromorphic ZW chromosomes and even multiple chromosome systems (Z1Z1Z2Z2/Z1Z2W). To gain a deeper insight into the patterns of karyotype and sex chromosome evolution, we performed a cytogenetic analysis based on conventional staining, banding techniques and fluorescence in situ hybridization in the species I. monticola, for which previous cytogenetic investigations did not detect differentiated sex chromosomes. The karyotype is composed of 2n = 36 acrocentric chromosomes. NORs and the major ribosomal genes were located in the subtelomeric region of chromosome pair 6. Hybridization signals of the telomeric sequences (TTAGGG)n were visualized at the telomeres of all chromosomes and interstitially in 5 chromosome pairs. C-banding showed constitutive heterochromatin at the centromeres of all chromosomes, as well as clear pericentromeric and light telomeric C-bands in several chromosome pairs. These results highlight some chromosomal markers which can be useful to identify species-specific diagnostic characters, although they may not accurately reflect the phylogenetic relationships among the taxa. In addition, C-banding revealed the presence of a heteromorphic ZW sex chromosome pair, where W is smaller than Z and almost completely heterochromatic. This finding sheds light on sex chromosome evolution in the genus Iberolacerta and suggests that further comparative cytogenetic analyses are needed to understand the processes underlying the origin, differentiation and plasticity of sex chromosome systems in lacertid lizards.


Near-field dosimetry for in vitro exposure of human cells at 60 GHz.
Due to the expected mass deployment of millimeter-wave wireless technologies, thresholds of potential millimeter-wave-induced biological and health effects should be carefully assessed. The main purpose of this study is to propose, optimize, and characterize a near-field exposure configuration allowing illumination of cells in vitro at 60 GHz with power densities up to several tens of mW/cm(2) . Positioning of a tissue culture plate containing cells has been optimized in the near-field of a standard horn antenna operating at 60 GHz. The optimal position corresponds to the maximal mean-to-peak specific absorption rate (SAR) ratio over the cell monolayer, allowing the achievement of power densities up to 50 mW/cm(2) at least. Three complementary parameters have been determined and analyzed for the exposed cells, namely the power density, SAR, and temperature dynamics. The incident power density and SAR have been computed using the finite-difference time-domain (FDTD) method. The temperature dynamics at different locations inside the culture medium are measured and analyzed for various power densities. Local SAR, determined based on the initial rate of temperature rise, is in a good agreement with the computed SAR (maximal difference of 5%). For the optimized exposure setup configuration, 73% of cells are located within the ±3 dB region with respect to the average SAR. It is shown that under the considered exposure conditions, the maximal power density, local SAR, and temperature increments equal 57 mW/cm(2) , 1.4 kW/kg, and 6 °C, respectively, for the radiated power of 425 mW.


Cryopreservation of human mucosal tissues.
Cryopreservation of leukocytes isolated from the cervicovaginal and colorectal mucosa is useful for the study of cellular immunity (see Hughes SM et al. PLOS ONE 2016). However, some questions about mucosal biology and sexually transmitted infections are better addressed with intact mucosal tissue, for which there is no standard cryopreservation protocol. To find an optimal preservation protocol for mucosal tissues, we tested slow cooling (1°C/min) with 10% dimethylsulfoxide (designated "cryopreservation") and fast cooling (plunge in liquid nitrogen) with 20% dimethylsulfoxide and 20% ethylene glycol ("vitrification"). We compared fresh and preserved human cervicovaginal and colorectal tissues in a range of assays, including metabolic activity, human immunodeficiency virus infection, cell phenotype, tissue structure by hematoxylin-and-eosin staining, cell number and viability, production of cytokines, and microbicide drug concentrations. Metabolic activity, HIV infectability, and tissue structure were similar in cryopreserved and vitrified vaginal tissues. However, vitrification led to poor cell recovery from the colorectal mucosa, with 90% fewer cells recovered after isolation from vitrified colorectal tissues than from cryopreserved. HIV infection rates were similar for fresh and cryopreserved ectocervical tissues, whereas cryopreserved colorectal tissues were less easily infected than fresh tissues (hazard ratio 0.7 [95% confidence interval 0.4, 1.2]). Finally, we compared isolation of cells before and after cryopreservation. Cell recoveries were higher when cells were isolated after freezing and thawing (71% [59-84%]) than before (50% [38-62%]). Cellular function was similar to fresh tissue in both cases. Microbicide drug concentrations were lower in cryopreserved explants compared to fresh ones. Cryopreservation of intact cervicovaginal and colorectal tissues with dimethylsulfoxide works well in a range of assays, while the utility of vitrification is more limited. Cell yields are higher from cryopreserved intact tissue pieces than from thawed cryopreserved single cell suspensions isolated before freezing, but T cell functions are similar.


Identification of novel nitroreductases from Bacillus cereus and their interaction with the CB1954 prodrug.
Directed enzyme prodrug therapy is a form of cancer chemotherapy in which bacterial prodrug-activating enzymes, or their encoding genes, are directed to the tumour before administration of a prodrug. The prodrug can then be activated into a toxic drug at the tumour site, reducing off-target effects. The bacterial nitroreductases are a class of enzymes used in this therapeutic approach and although very promising, the low turnover rate of prodrug by the most studied nitroreductase enzyme, NfnB from Escherichia coli (NfnB_Ec), is a major limit to this technology. There is a continual search for enzymes with greater efficiency, and as part of the search for more efficient bacterial nitroreductase enzymes, two novel enzymes from Bacillus cereus (strain ATCC 14579) have been identified and shown to reduce the CB1954 (5-(aziridin-1-yl)-2,4-dinitrobenzamide) prodrug to its respective 2'-and 4'-hydroxylamine products. Both enzymes shared features characteristic of the nitro-FMN-reductase superfamily including non-covalently associated FMN, requirement for the NAD(P)H cofactor, homodimeric, could be inhibited by Dicoumarol (3,3'-methylenebis(4-hydroxy-2H-chromen-2-one)), and displayed ping pong bi bi kinetics. Based on the biochemical characteristics and nucleotide alignment with other nitroreductase enzymes, one enzyme was named YdgI_Bc and the other YfkO_Bc. Both B. cereus enzymes had greater turnover for the CB1954 prodrug compared with NfnB_Ec, and in the presence of added NADPH cofactor, YfkO_Bc had superior cell killing ability, and produced mainly the 4'-hydroxylamine product at low prodrug concentration. The YfkO_Bc was identified as a promising candidate for future enzyme prodrug therapy.


[Congenital thrombocytopenia in adults].
Thrombocytopenia in adult patients is usually acquired. Important causes are medication, pregnancy, infections and auto-immune disorders. Here we present 2 patients, a 32-year-old and a 45-year-old man, in whom congenital thrombocytopenia was discovered in adulthood, caused by a mutation in the gene for Wiskott-Aldrich syndrome. This congenital thrombocytopenia appears in different forms. Indications for the disorder are chronic thrombocytopenia, a family history and additional clinical signs such as eczema and renal insufficiency. Measurement of the thrombocyte size is a helpful tool in the diagnosis. Recognition of congenital thrombocytopenia can have important clinical consequences.


Influence of Altitude on Phytochemical Composition of Hemp Inflorescence: A Metabolomic Approach.
The phytochemical profiling of hemp inflorescences of clonal plants growing in different conditions related to altitude was investigated. Four strains of industrial hemp (Cannabis sativa L., family Cannabaceae) of Kompolti variety were selected and cloned to provide genetically uniform material for analyses of secondary metabolites (terpenes, cannabinoids, and flavonoids) at two different elevations: mountain (Alagna Valsesia 1200 m ASL) and plains (Vercelli Province 130 m ASL). Environmental conditions influenced by elevation have proven to be important factors inducing variations in hemp inflorescences' secondary metabolite composition. In fact, all plants grown at altitude exhibited a higher total amount of terpenes when compared with plains counterparts, with β-Myrcene, trans-Caryophyllene and α-Humulene as the main contributors. A metabolomic, un-targeted approach performed by HPLC-Q-Exactive-Orbitrap®-MS platform with subsequent data processing performed by Compound Discoverer™ software, was crucial for the appropriate recognition of many metabolites, clearly distinguishing mountain from plains specimens. Cannabidiolic acid CBDA was the most abundant phytocannabinoid, with significantly higher concentrations in the mountain samples. The metabolic pathway of CBGA (considered as the progenitor/precursor of all cannabinoids) was also activated towards the production of CBCA, which occurs in considerably 3 times higher quantities than in the clones grown at high altitude. Isoprenoid flavones (Cannaflavins A, B, and C) were correspondingly upregulated in mountain samples, while apigenin turned out to be more abundant in plains samples. The possibility to use hemp inflorescences in pharmaceutical/nutraceutical applications opens new challenges to understand how hemp crops respond in terms of secondary metabolite production in various environments. In this regard, our results with the applied analytical strategy may constitute an effective way of phytochemical profiling hemp inflorescences.


Emerging technologies for the pretreatment of lignocellulosic biomass.
Pretreatment of lignocellulosic biomass to overcome its intrinsic recalcitrant nature prior to the production of valuable chemicals has been studied for nearly 200 years. Research has targeted eco-friendly, economical and time-effective solutions, together with a simplified large-scale operational approach. Commonly used pretreatment methods, such as chemical, physico-chemical and biological techniques are still insufficient to meet optimal industrial production requirements in a sustainable way. Recently, advances in applied chemistry approaches conducted under extreme and non-classical conditions has led to possible commercial solutions in the marketplace (e.g. High hydrostatic pressure, High pressure homogenizer, Microwave, Ultrasound technologies). These new industrial technologies are promising candidates as sustainable green pretreatment solutions for lignocellulosic biomass utilization in a large scale biorefinery. This article reviews the application of selected emerging technologies such as ionizing and non-ionizing radiation, pulsed electrical field, ultrasound and high pressure as promising technologies in the valorization of lignocellulosic biomass.


Modeling Lyme disease host animal habitat suitability, West Point, New York.
As the most frequently reported vector-borne disease among active component U.S. service members, with an incidence rate of 16 cases per 100,000 person-years in 2011, Lyme disease poses both a challenge to healthcare providers in the Military Health System and a threat to military readiness. Spread through the bite of an infected blacklegged tick, infection with the bacterial cause of Lyme disease can have lasting effects that may lead to medical discharge from the military. The U.S. Military Academy at West Point is situated in a highly endemic area in New York State. To identify probable areas where West Point cadets as well as active duty service members stationed at West Point and their families might contract Lyme disease, this study used Geographic Information System mapping methods and remote sensing data to replicate an established spatial model to identify the likely habitat of a key host animal--the white-tailed deer.


Quantum-confined single photon emission at room temperature from SiC tetrapods.
Controlled engineering of isolated solid state quantum systems is one of the most prominent goals in modern nanotechnology. In this letter we demonstrate a previously unknown quantum system namely silicon carbide tetrapods. The tetrapods have a cubic polytype core (3C) and hexagonal polytype legs (4H)--a geometry that creates spontaneous polarization within a single tetrapod. Modeling of the tetrapod structures predicts that a bound exciton should exist at the 3C-4H interface. The simulations are confirmed by the observation of fully polarized and narrowband single photon emission from the tetrapods at room temperature. The single photon emission provides important insights into understanding the quantum confinement effects in non-spherical nanostructures. Our results pave the way to a new class of crystal phase nanomaterials that exhibit single photon emission at room temperature and therefore are suitable for sensing, quantum information and nanophotonics.


Assessment of flower number per inflorescence in grapevine by image analysis under field conditions.
Flowers, flowering and fruit set are key determinants of grapevine yield. Currently, practical methods to assess the flower number per inflorescence, necessary for fruit set estimation, are time and labour demanding. This work aims at developing a simple, cheap, fast, accurate and robust machine vision methodology to be applied to RGB images taken under field conditions, to estimate the number of flowers per inflorescence automatically. Ninety images of individual inflorescences of Vitis vinifera L. cultivars Tempranillo, Graciano and Carignan were acquired in the vineyard with a pocket RGB camera prior to flowering, and used to develop and test the 'flower counting' algorithm. Strong and significant relationships, with R(2) above 80% for the three cultivars were observed between actual and automated estimation of inflorescence flower numbers, with a precision exceeding 90% for all cultivars. The developed algorithm proved that the analysis of digital images captured by pocket cameras under uncontrolled outdoors conditions was able to automatically provide a useful estimation of the number of flowers per inflorescence of grapevines at early stages of flowering.


Anatomic changes related to laryngeal descent from birth to 1 year of age: do they play a role in SIDS?
We conducted a retrospective study to measure laryngeal descent in human infants and to determine if there is any correlation between the associated anatomic changes and the timing of the peak incidence of sudden infant death syndrome (SIDS), which is 2 to 4 months of age. We performed a computerized search of hospital records at our institution to identify magnetic resonance imaging (MRI) scans of the head and neck and plain radiographs of the lateral neck in patients younger than 1 year of age (range: 1 to 357 days). After unusable images were excluded, 79 head and neck MRI scans and 111 lateral neck x-rays were suitable for study. Two measurements were taken from each image: one from the tip of the epiglottis to the uvula and one from the tip of the epiglottis to the center of the sella turcica. These measurements were then graphed against the subject's age. SPSS statistical software was used to determine growth curves of the various measurements. The first derivative of these curves was calculated to determine the rate of laryngeal descent at a given age. We found that most subjects did not have an overlapping epiglottis and uvula during the first few months of life. The rate of laryngeal descent, based on measurements of the distance between the epiglottis and uvula, gradually increased in a near-linear fashion from as low as 0.005726 mm/day at day 1 of life to as high as 0.028366 mm/day at 300 days of age. We found no sharp increase in the rate of descent at 2 to 4 months of age, and thus no support for our hypothesis that there might be a correlation between anatomic changes and the peak incidence of SIDS.


Genome-wide maps of nuclear lamina interactions in single human cells.
Mammalian interphase chromosomes interact with the nuclear lamina (NL) through hundreds of large lamina-associated domains (LADs). We report a method to map NL contacts genome-wide in single human cells. Analysis of nearly 400 maps reveals a core architecture consisting of gene-poor LADs that contact the NL with high cell-to-cell consistency, interspersed by LADs with more variable NL interactions. The variable contacts tend to be cell-type specific and are more sensitive to changes in genome ploidy than the consistent contacts. Single-cell maps indicate that NL contacts involve multivalent interactions over hundreds of kilobases. Moreover, we observe extensive intra-chromosomal coordination of NL contacts, even over tens of megabases. Such coordinated loci exhibit preferential interactions as detected by Hi-C. Finally, the consistency of NL contacts is inversely linked to gene activity in single cells and correlates positively with the heterochromatic histone modification H3K9me3. These results highlight fundamental principles of single-cell chromatin organization. VIDEO ABSTRACT.


An inverse relationship to germline transcription defines centromeric chromatin in C. elegans.
Centromeres are chromosomal loci that direct segregation of the genome during cell division. The histone H3 variant CENP-A (also known as CenH3) defines centromeres in monocentric organisms, which confine centromere activity to a discrete chromosomal region, and holocentric organisms, which distribute centromere activity along the chromosome length. Because the highly repetitive DNA found at most centromeres is neither necessary nor sufficient for centromere function, stable inheritance of CENP-A nucleosomal chromatin is postulated to propagate centromere identity epigenetically. Here, we show that in the holocentric nematode Caenorhabditis elegans pre-existing CENP-A nucleosomes are not necessary to guide recruitment of new CENP-A nucleosomes. This is indicated by lack of CENP-A transmission by sperm during fertilization and by removal and subsequent reloading of CENP-A during oogenic meiotic prophase. Genome-wide mapping of CENP-A location in embryos and quantification of CENP-A molecules in nuclei revealed that CENP-A is incorporated at low density in domains that cumulatively encompass half the genome. Embryonic CENP-A domains are established in a pattern inverse to regions that are transcribed in the germline and early embryo, and ectopic transcription of genes in a mutant germline altered the pattern of CENP-A incorporation in embryos. Furthermore, regions transcribed in the germline but not embryos fail to incorporate CENP-A throughout embryogenesis. We propose that germline transcription defines genomic regions that exclude CENP-A incorporation in progeny, and that zygotic transcription during early embryogenesis remodels and reinforces this basal pattern. These findings link centromere identity to transcription and shed light on the evolutionary plasticity of centromeres.


The age-specific quantitative effects of metabolic risk factors on cardiovascular diseases and diabetes: a pooled analysis.
The effects of systolic blood pressure (SBP), serum total cholesterol (TC), fasting plasma glucose (FPG), and body mass index (BMI) on the risk of cardiovascular diseases (CVD) have been established in epidemiological studies, but consistent estimates of effect sizes by age and sex are not available. We reviewed large cohort pooling projects, evaluating effects of baseline or usual exposure to metabolic risks on ischemic heart disease (IHD), hypertensive heart disease (HHD), stroke, diabetes, and, as relevant selected other CVDs, after adjusting for important confounders. We pooled all data to estimate relative risks (RRs) for each risk factor and examined effect modification by age or other factors, using random effects models. Across all risk factors, an average of 123 cohorts provided data on 1.4 million individuals and 52,000 CVD events. Each metabolic risk factor was robustly related to CVD. At the baseline age of 55-64 years, the RR for 10 mmHg higher SBP was largest for HHD (2.16; 95% CI 2.09-2.24), followed by effects on both stroke subtypes (1.66; 1.39-1.98 for hemorrhagic stroke and 1.63; 1.57-1.69 for ischemic stroke). In the same age group, RRs for 1 mmol/L higher TC were 1.44 (1.29-1.61) for IHD and 1.20 (1.15-1.25) for ischemic stroke. The RRs for 5 kg/m(2) higher BMI for ages 55-64 ranged from 2.32 (2.04-2.63) for diabetes, to 1.44 (1.40-1.48) for IHD. For 1 mmol/L higher FPG, RRs in this age group were 1.18 (1.08-1.29) for IHD and 1.14 (1.01-1.29) for total stroke. For all risk factors, proportional effects declined with age, were generally consistent by sex, and differed by region in only a few age groups for certain risk factor-disease pairs. Our results provide robust, comparable and precise estimates of the effects of major metabolic risk factors on CVD and diabetes by age group.


Global challenges in preparedness and response to epidemic infectious diseases.
Lessons drawn from successes and failures with coronavirus disease 2019 (COVID-19) and Ebola virus disease (EVD) should help shaping a robust health innovation system for infectious disease epidemics. Epidemic response research and development (R&D) can be mobilized quickly for public health priorities and yield medicinal products within months. However, to resolve epidemics, technological advances must be equitably accessible and deployed, and these examples expose the limitations of a supply-driven, fragmented R&D ecosystem relying primarily on the private sector to deliver health products. Efficient epidemic response requires a coordinated public health-focused, end-to-end R&D ecosystem for the development, registration, availability, and use of pharmaceutical products. Because pivotal clinical trials can only be conducted during outbreaks, significant preparation must be done beforehand: strengthening clinical research capacity and developing pre-positioned trial protocols and clinical characterization protocols, as well as conducting discovery and pre-clinical research, manufacturing, and early clinical testing of candidate products. This will allow for speedy execution of clinical research early into an outbreak and delivering products within a short time. Effective interventions should be adopted and deployed ensuring equitable access during the ongoing outbreak. Measures to make products available where and when needed must be integrated throughout the R&D value chain.


Analytical properties of the plasmon decay profile in a periodic metal-nanoparticle chain.
We show that the spatial decay of plasmons in a periodic metal-nanoparticle chain is composed of exponential and power-law decays. Our results show a high level of similarity between the absorptive and radiative decay channels. By analyzing the poles (and the corresponding residues) of the generating function for the lattice Green's function, we explain the details of the spatial decay profile. We also present an analytical formula for the decay profile.


Current perspectives and emerging issues on cancer rehabilitation.
Cancer rehabilitation is a rapidly emerging and evolving medical field in both Europe and the United States, in large part because of increases in the number of cancer survivors. Although few argue with the need to restore function and quality of life to patients affected by cancer and its treatments, differences exist between European countries with regard to the funding, accessibility, and even the definition of cancer rehabilitation services. In the United States, there is tremendous variability in the provision of rehabilitation services resulting from a variety of factors, including a lack of highly trained cancer rehabilitation physicians and therapists as well as a lack of comprehensive cancer rehabilitation programs, even at the majority of top cancer centers. Although studies evaluating the effectiveness of rehabilitation programs in the cancer setting, particularly exercise, have influenced clinical decision-making in both Europe and the United States for some time, this emerging evidence base also is now starting to influence guideline and policy making. Coordinated research efforts are essential to establish a robust framework to support future investigation and establish shared initiatives. Determining the best way forward for cancer survivors will require investment in large-scale prospective cohort studies that sufficiently describe their rehabilitation needs through the continuum of the survivorship experience.


Ten Rules for Conducting Retrospective Pharmacoepidemiological Analyses: Example COVID-19 Study.
Since the beginning of the COVID-19 pandemic, pharmaceutical treatment hypotheses have abounded, each requiring careful evaluation. A randomized controlled trial generally provides the most credible evaluation of a treatment, but the efficiency and effectiveness of the trial depend on the existing evidence supporting the treatment. The researcher must therefore compile a body of evidence justifying the use of time and resources to further investigate a treatment hypothesis in a trial. An observational study can provide this evidence, but the lack of randomized exposure and the researcher's inability to control treatment administration and data collection introduce significant challenges. A proper analysis of observational health care data thus requires contributions from experts in a diverse set of topics ranging from epidemiology and causal analysis to relevant medical specialties and data sources. Here we summarize these contributions as 10 rules that serve as an end-to-end introduction to retrospective pharmacoepidemiological analyses of observational health care data using a running example of a hypothetical COVID-19 study. A detailed supplement presents a practical how-to guide for following each rule. When carefully designed and properly executed, a retrospective pharmacoepidemiological analysis framed around these rules will inform the decisions of whether and how to investigate a treatment hypothesis in a randomized controlled trial. This work has important implications for any future pandemic by prescribing what we can and should do while the world waits for global vaccine distribution.


Genetic evaluations for mixed-breed populations.
An all-breed animal model was developed for routine genetic evaluations of US dairy cattle. Data sets from individual breeds were combined, and records from crossbred cows were included. About 1% of recent cows were first-generation crossbreds. The numbers of cows with records since 1960 ranged from 10 to 22 million for the 6 traits analyzed, which were milk, fat, protein, somatic cell score, productive life, and daughter pregnancy rate. Programs were modified to account for general heterosis, to group unknown parents separately by breed, to adjust variances separately by breed, and to adjust data to a 36-mo age equivalent instead of a mature equivalent. Convergence rate of the all-breed model was similar to that of the previous within-breed animal model. Estimated breed differences were similar to those obtained previously from phenotypic breed means or from studies of crossbred cows and their herd-mates. Genetic evaluations from the all-breed and within-breed systems had high correlations: >0.99 for recent Holsteins and slightly <0.99 for other breeds. Predicted transmitting abilities will be converted back to the within-breed bases for purebred animals and to the breed of sire base for crossbred animals so that most purebred breeders will not be affected by the change to a multibreed model. Evaluations of crossbred animals from the multibreed model can include accurate information for both parents. Reliabilities also increase for purebred relatives because of the additional crossbred records and in mixed breed herds because cows of other breeds are additional contemporaries. Another benefit of the multibreed model is that breed differences are routinely estimated and updated. More research and education may be needed on using the new evaluations in the design of breeding programs. Implementation is expected in May 2007.