Group A rotavirus veterinary vaccines.
Group A rotaviruses cause diarrhea in young livestock and poultry; consequently, vaccination strategies have focused on induction of active or passive immunity. Gnotobiotic pigs and calves serve as useful models to evaluate induction of active immunity by candidate animal or human rotavirus vaccines. However, live attenuated rotavirus vaccines lacked efficacy when administered orally to calves and pigs in the field, presumably because colostral antibodies inhibited vaccine virus replication. The widespread occurrence of rotavirus antibodies in colostrum led to strategies for maternal rotavirus vaccination to boost lactogenic immunity and transfer passive antibodies to the neonate via colostrum and milk. The variable success of maternal rotavirus vaccines in the field is influenced by vaccine dose, strain, inactivating agent, adjuvant, route of administration, and environmental rotavirus exposure levels. The use of genetically engineered rotavirus-like particle vaccines in cows to boost antibodies in mammary secretions shows promise. Such subunit vaccines possess potential advantages over existing vaccines.


A systematic assessment of the quality of reporting for platelet transfusion studies.
As evidence-based medicine assumes increasing importance, there is a need for high-quality reporting of clinical studies. A recent review of clinical platelet (PLT) studies indicated variability in reporting. We undertook a critical analysis of PLT transfusion studies to determine the quality of reporting. A systematic MEDLINE search for clinical studies of PLT transfusion was performed to identify articles. Relevant observational studies (OBS) were critiqued using the STROBE checklist and randomized controlled clinical trials (RCTs) using the CONSORT checklist. Studies were further evaluated with a PLT-specific checklist developed by the authors. Observations were analyzed descriptively and using Pareto analysis. A total of 772 articles were identified by the search. Eighty-six articles (23 RCTs and 63 OBS) met eligibility criteria. All RCTs, and a similar number of OBS (24), were randomly selected for analysis. Studies reported the scientific background and rationale, key results, and outcomes. OBS frequently did not consider bias and confounders. RCTs frequently did not explain bias, interim analyses, stopping rules, success of blinding, or weaknesses of multiple analyses. The PLT-specific critique found many studies adequately reported basics of the PLT product, PLT increment, and transfusion reactions. Studies frequently failed to report specific details of PLT compatibility, details of product preparation, and use of other blood products. Recently published articles of clinical PLT transfusion share common strengths and weaknesses. The quality of reporting may be improved by providing guidelines to authors and journal editors that list the essential elements of a well-reported clinical study of PLT transfusion.


Effect of phonological and morphological awareness on reading comprehension in Hebrew-speaking adolescents with reading disabilities.
This research explored phonological and morphological awareness among Hebrew-speaking adolescents with reading disabilities (RD) and its effect on reading comprehension beyond phonological and word-reading abilities. Participants included 39 seventh graders with RD and two matched control groups of normal readers: 40 seventh graders matched for chronological age (CA) and 38 third graders matched for reading age (RA). We assessed phonological awareness, word reading, morphological awareness, and reading comprehension. Findings indicated that the RD group performed similarly to the RA group on phonological awareness but lower on phonological decoding. On the decontextualized morphological task, RD functioned on par with RA, whereas in a contextualized task RD performed above RA but lower than CA. In reading comprehension, RD performed as well as RA. Finally, results indicated that for normal readers contextual morphological awareness uniquely contributed to reading comprehension beyond phonological and word-reading abilities, whereas no such unique contribution emerged for the RD group. The absence of an effect of morphological awareness in predicting reading comprehension was suggested to be related to a different recognition process employed by RD readers which hinder the ability of these readers to use morphosemantic structures. The lexical quality hypothesis was proposed as further support to the findings, suggesting that a low quality of lexical representation in RD students leads to ineffective reading skills and comprehension. Lexical representation is thus critical for both lexical as well as comprehension abilities.


Psychometric properties of the Dutch revised sense of coherence scale in a firefighter sample.
Sense of coherence (SOC) has been associated with resilience to posttraumatic stress disorder (PTSD) and seems to be a promising factor in primary prevention of PTSD in high risk populations. The present study evaluated the psychometric properties of the Dutch revised Sense of Coherence Scale (SOC-R) in a sample of N = 527 firefighters. To investigate the internal structure of this 13-item scale, a Mokken scale analysis and an exploratory factor analysis were conducted (i.e. parallel analysis based on MRFA). The combined results of these analyses suggested that a one-factor solution with 10 out of 13 items was most compelling for our firefighter sample. Reliability estimates for the 10-item version increased compared to the 13-item version (13-item: α = .82, λ2 = .83; 10-item: α = λ2 = .85). As expected, the Dutch version showed positive associations with resilience (convergent validity), and low correlations with neuroticism and extraversion (discriminant validity). The one-factor solution of the Dutch SOC-R with 10 items (excluding item 2, item 3, and item 6) is most convincing. The use of this scale might be specifically interesting regarding its potential to primary prevention of trauma-related psychopathology in high-risk samples.


How the anal gland orifice could be found in anal abscess operations.
On an average 30-50% of patients who undergo incision and drainage (I and D) of anal abscess will develop recurrence or fistula formation. It is claimed that finding the internal orifice of anal abscess to distract the corresponding anal gland duct; will decline the rate of future anal fistula. Surgeons supporting I and D alone claim that finding the internal opening is hazardous. This study is conducted to assess short-term results of optional method to manage patients with anal abscess and fitula-in-ano at the same time. In this cross-sectional descriptive study 49 from 77 patients with anal abscess whose internal orifice was not identified by pressing on the abscess, diluted hydrogen peroxide (2%) and methylene blue was injected into the abscess cavity and the anal canal was inspected to find out the internal opening. Once the opening was distinguished, an incision was given from the anal verge to the internal opening. The internal orifice was identified in 44 out of 49 patients (90%) who underwent this new technique. Up to 18 months during follow-up, only 2.5% of patients with primary fistulotomy developed fistula on the site of a previous abscess. Conventional method to seek the internal orifice of anal abscesses is successful in about one-third of cases. By applying this new technique, surgeons would properly find the internal opening in >90% of patients. Needless to say, safe identification of the anal gland orifice in anal abscess disease best helps surgeons to do primary fistulotomy and in turn it would significantly decrease the rate of recurrence in anal abscess and fistula formation.


Structural study by solid-state (71)Ga NMR of thin film transistor precursors.
Solid-state (71)Ga NMR was used to investigate the structures of several heterometallic Group 13 hydroxo-aquo clusters, [Ga13-xInx (μ3-OH)6(μ2-OH)18(H2O)24](NO3)15 which are envisioned for thin film transistors. The characterization of these clusters in the solid state provides additional information in understanding the synthesis, structure and speciation of these precursors for high-quality, ultrasmooth thin films. Yet important structural information regarding these clusters - including the exact composition, isomeric structure, and coordination environments - were unknown prior to this precise NMR spectroscopy study. These molecular species, termed "Ga13-xInx", contain three types of six-coordinate metal sites, with bridging OH(-) groups and H2O as capping ligands, and we report results on Ga7In6, Ga8In5, Ga10In3, Ga11In2, Ga12In1, and Ga13. Utilizing two magnetic fields (13.9 T and 21.1 T), the solid-state NMR spectra were interpreted in conjunction with computational modeling (using CASTEP) and simulation of spectral lineshapes (using Dmfit). The metal sites are best represented as distorted octahedra, and they exhibit a range of quadrupolar couplings and asymmetry parameters, which can be addressed using longitudinal strain analysis. Until now, there has been speculation about the sites for transmetallation within the synthetic cluster community. Here, we show that Ga NMR is a powerful technique to monitor the transmetallation of In for Ga in the Ga13-xInx clusters, specifically substituting in the "outer ring" sites, and not the "core" or "middle ring".


Preoperative predictors of persistent impairments during stair ascent and descent after total knee arthroplasty.
Although total knee arthroplasty improves functional mobility in persons with end-stage knee osteoarthritis, many subjects have reported continued difficulty with stair ascent and descent after surgery. The purpose of the present study was to determine preoperative predictors of handrail use during stair ascent and descent following primary unilateral total knee arthroplasty. One hundred and five adults who were scheduled for unilateral total knee arthroplasty participated in the study. Postoperative handrail use during stair ascent or descent was predicted on the basis of preoperative functional measures. Preoperative age, body mass index, knee strength, knee flexion active range of motion, Knee Outcome Survey scores, time to complete a stair-climbing task, and previous handrail use were entered as covariates into a binary logistic regression. Forward logistic regression was performed to determine which preoperative factors best predicted handrail use at three months and two years after surgery. Handrail use in a control group was also evaluated at baseline and at the time of the two-year follow-up. Prior to surgery, sixty-three of the 105 subjects required a handrail. Two years after surgery, sixty of the 105 subjects required a handrail. In the control group, nineteen of the sixty-four subjects required a handrail at baseline and ten of thirty-one required a handrail at the time of the two-year follow-up. At two years, the preoperative ability to ascend and descend stairs without a handrail was the best predictor of individuals who would not require a handrail after surgery, followed by younger age and greater quadriceps strength. Collectively, these variables correctly predicted the ability of ninety of 105 persons to negotiate stairs without a handrail at two years after surgery (p < 0.001). Younger, stronger patients who do not use a handrail prior to unilateral total knee arthroplasty can expect the best outcomes in terms of ascending and descending stairs following surgery. This information may provide patients with more realistic expectations after surgery and allow them to make more appropriate discharge plans.


Phenomenological modeling of durotaxis.
Cells exhibit qualitatively different behaviors on substrates with different rigidities. The fact that cells are more polarized on the stiffer substrate motivates us to construct a two-dimensional cell with the distribution of focal adhesions dependent on substrate rigidities. This distribution affects the forces exerted by the cell and thereby determines its motion. Our model reproduces the experimental observation that the persistence time is higher on the stiffer substrate. This stiffness-dependent persistence will lead to durotaxis, the preference in moving towards stiffer substrates. This propensity is characterized by the durotaxis index first defined in experiments. We derive and validate a two-dimensional corresponding Fokker-Planck equation associated with our model. Our approach highlights the possible role of the focal adhesion arrangement in durotaxis.


Complementary and alternative medicine: use and disclosure in radiation oncology community practice.
The aims of this study were to evaluate the frequency of complementary and alternative medicine (CAM) use among radiation oncology patients, the coping strategies that influenced this use, and the rates of disclosure of CAM use to their healthcare providers. One hundred fifty-three patients undergoing radiation therapy for various neoplasms at rural cancer centers in Minnesota completed the Mayo Complementary and Alternative Medicine Use Survey and the Coping Inventory for Stressful Situations questionnaires. Data regarding CAM use was also compared with provider consultation notes in the medical record at the onset of radiation therapy to determine rates of patient disclosure of CAM use to their healthcare providers. A total of 153 participants completed the study with 61.4% females and 38.6% males and a mean age of 64.9 years. The two most frequent diagnoses of participants were breast cancer (43.8%) and prostate cancer (22.9%). CAM use was reported in 95% of the participants and was categorized into three domains: treatments and techniques, vitamins, and herbs and supplements. The three most frequently reported treatments and techniques were spiritual healing/prayer (62.1%), exercise (19.6%), and music (17.6%). The top three most frequently used biologically based CAM therapies were multivitamins (48.1%), calcium (37.3%), and vitamin with minerals (21.5%). The most frequently used herbs and other dietary supplements were fish oil (19.0%), flaxseed (15.0%), glucosamine (15.0%), and green tea (15.0%). The most common reason cited for CAM treatments and techniques use was previous use (26.1%), for use of vitamins and minerals was recommendation by a physician (33.0%), and for use of herbs and other supplements was previous use (19.0%). One hundred twelve participants reported taking vitamins, minerals, or supplements, and 47% of those 112 did not disclose this use to their providers. Consistent with previous research, our study found that the majority of cancer patients used CAM treatments. Spiritual healing/prayer was the most commonly reported, followed by multivitamins. Patients reported using CAM primarily due to previous use and physician recommendation. Unfortunately, disclosure of CAM use to healthcare providers was relatively low.


Calcium metabolism in health and disease.
This brief review focuses on calcium balance and homeostasis and their relationship to dietary calcium intake and calcium supplementation in healthy subjects and patients with chronic kidney disease and mineral bone disorders (CKD-MBD). Calcium balance refers to the state of the calcium body stores, primarily in bone, which are largely a function of dietary intake, intestinal absorption, renal excretion, and bone remodeling. Bone calcium balance can be positive, neutral, or negative, depending on a number of factors, including growth, aging, and acquired or inherited disorders. Calcium homeostasis refers to the hormonal regulation of serum ionized calcium by parathyroid hormone, 1,25-dihydroxyvitamin D, and serum ionized calcium itself, which together regulate calcium transport at the gut, kidney, and bone. Hypercalcemia and hypocalcemia indicate serious disruption of calcium homeostasis but do not reflect calcium balance on their own. Calcium balance studies have determined the dietary and supplemental calcium requirements needed to optimize bone mass in healthy subjects. However, similar studies are needed in CKD-MBD, which disrupts both calcium balance and homeostasis, because these data in healthy subjects may not be generalizable to this patient group. Importantly, increasing evidence suggests that calcium supplementation may enhance soft tissue calcification and cardiovascular disease in CKD-MBD. Further research is needed to elucidate the risks and mechanisms of soft tissue calcification with calcium supplementation in both healthy subjects and CKD-MBD patients.


A statistical method for quantifying songbird phonology and syntax.
Songbirds are the preeminent animal model for understanding how the brain encodes and produces learned vocalizations. Here, we report a new statistical method, the Kullback-Leibler (K-L) distance, for analyzing vocal change over time. First, we use a computerized recording system to capture all song syllables produced by birds each day. Sound Analysis Pro software [Tchernichovski O, Nottebohm F, Ho CE, Pesaran B, Mitra PP. A procedure for an automated measurement of song similarity. Anim Behav 2000;59:1167-76] is then used to measure the duration of each syllable as well as four spectral features: pitch, entropy, frequency modulation, and pitch goodness. Next, two-dimensional scatter plots of each day of singing are created where syllable duration is on the x-axis and each of the spectral features is represented separately on the y-axis. Each point in the scatter plots represents one syllable and we regard these plots as random samples from a probability distribution. We then apply the standard information-theoretic quantity K-L distance to measure dissimilarity in phonology across days of singing. A variant of this procedure can also be used to analyze differences in syllable syntax.


Manuka honey incorporated cellulose acetate nanofibrous mats: Fabrication and in vitro evaluation as a potential wound dressing.
Wound dressings are the primary barrier between the wound surface and the outer environment. Here we report the fabrication of cellulose acetate (CA)-Manuka honey (MH) composite nanofibrous mats as a biocompatible and antimicrobial wound dressing. CA mats with different quantities of MH were developed by electrospinning. The ATR-FTIR spectra confirm the inclusion of MH in the composite CA-MH nanofibrous mats. The fibers were continuous and bead-free with acceptable mechanical properties. The fiber diameter increased with an increase in MH content. Inclusion of MH in the electrospun composite CA-MH nanofibrous mats shows high efficacy to prevent bacterial growth on the wound surface. The MH loaded CA nanofiber mats showed good antioxidant abilities, while the ability to free radicalize the DPPH was dependent upon the factors of MH content in the fiber and the time of immersion in the DPPH solution. Besides, the nanofibrous mat's high porosity (85-90%) and WVTR values of 2600 to 1950 g/m2/day, suitable for wound breathability and the mats show high cytocompatibility to NIH 3T3 cell line in in vitro testing, proving to be effective for promoting wound healing.


Adult response to olanzapine or clozapine treatment is altered by adolescent antipsychotic exposure: a preclinical test in the phencyclidine hyperlocomotion model.
This study examined how repeated olanzapine (OLZ) or clozapine (CLZ) treatment in adolescence alters sensitivity to the same drug in adulthood in the phencyclidine (PCP) hyperlocomotion model. Male adolescent Sprague-Dawley rats (postnatal day (P) 44-48) were first treated with OLZ (1.0 or 2.0 mg/kg, subcutaneously (sc)) or CLZ (10.0 or 20.0 mg/kg, sc) and tested in the PCP (3.2 mg/kg, sc)-induced hyperlocomotion model for five consecutive days. Then a challenge test with OLZ (0.5 mg/kg) or CLZ (5.0 mg/kg) was administered either during adolescence (~P 51) or after the rats matured into adults (~P 76 and 91). During adolescence, repeated OLZ or CLZ treatment produced a persistent inhibition of PCP-induced hyperlocomotion across the five test days. In the challenge test during adolescence, rats previously treated with OLZ did not show a significantly stronger inhibition of PCP-induced hyperlocomotion than those previously treated with vehicle (VEH). In contrast, those previously treated with CLZ showed a weaker inhibition than the VEH controls. When assessed in adulthood, the enhanced sensitivity to OLZ and the decreased sensitivity to CLZ were detected on ~P 76, even on ~P 91 in the case of OLZ. These findings suggest that adolescent OLZ or CLZ exposure can induce long-term alterations in antipsychotic response that persist into adulthood.


[Influence of temperature and humidity on the nymphal development of Rhodnius robustus].
To assess the significant differences in the nymphal development of the Rhodnius robustus Larrousse, 1927 under different temperatures and humidity conditions. This is a species found in the northern region of Brazil (states of Acre, Amazonas, and Pará), Colombia, Equator, Peru and Venezuela. Three groups of triatominae were kept under the following laboratory conditions: 33/40 (33+1 degree C and 40+/-5% of relative humidity - RH), 33/70 (33+/-1 degree C and 70+/-5% RH), and 28/70 (28+/-1 degree C and 70+/-5% RH). The incubation period of the eggs, developmental time of each stage, mortality percentage, number of bloodmeals, and the total amount of time from the egg hatching to adult ecdysis were observed. The shortest average time of nymphal development was observed in the 28/70 group, with following averages: 14.4, 17.3, 20.3, 22.8, and 40. Significant differences were observed in the embryonic development between the groups (p<0.01). For all groups, the number of bloodmeals had a gradual increase near the adult phase, except for the 3rd instar of the 33/70 group. The smallest mortality percentages were seen in the 28/70 group. High temperatures, regardless of the humidity, can accelerate the biological development cycle of R. robustus. However, these temperatures can impair the colonies' survival, preventing their maintenance in the laboratory.


Acute and chronic aortic dissections--determinants of long-term outcome for operative survivors.
A total of 135 survivors of surgical treatment of aortic dissection was followed for up to 15 years after surgery. Actuarial survival rates were 82 +/- 4% at 5 years and 64 +/- 6% at 10 years. There were no significant differences in long-term survival rates of patients in four subsets based on type and acuity of dissections. The incidence of late reoperation (dissection-related) was 13 +/- 4% at 5 years and 23 +/- 6% at 10 years; again, there was no significant difference among patients with different types or acuity of dissection. Multiple variables were investigated by multivariate discriminant analysis. Significant independent risk factors for late death included stroke, chronic renal dysfunction, remote myocardial infarction, and operation in the early years of this study. Younger age, site of intimal tear (arch), and cardiac tamponade portended a significantly higher likelihood of late reoperation. Except for stroke, no complication of the dissection or intraoperative factor significantly influenced late survival. Patients in whom the intimal tear was located in the aortic arch had the highest probability of late reoperation. Thus, dissection type, acuity, and distal extent, whether or not the tear was resected or concomitant aortic valve replacement performed, and a host of patient-related characteristics had no significant influence on the generally excellent long-term prognosis after surgical treatment. Indefinite surveillance of the remaining natural aorta is imperative (with reoperation when indicated) to attain such results.


Interaction of Rotylenchulus reniformis with Seedling Disease Pathogens of Cotton.
The impact of 10 Fusarium species in concomitant association with Rotylenchulus reniformis on cotton seedling disease was examined under greenhouse conditions. In experiment 1, fungal treatments consisted of Fusarium chlamydosporum, F. equiseti, F. lateritium, F. moniliforme, F. oxysporum, F. oxysporum f.sp. vasinfectum, F. proliferatum, F. semitectum, F. solani, and F. sporotrichioides; Rhizoctonia solani; and Thielaviopsis basicola. The experimental design was a 2 x 14 factorial consisting of the presence or absence of R. reniformis and the 12 fungal treatments plus two controls in autoclaved field soil. In experiment 2, the same fungal and nematode treatments were examined in autoclaved or non-autoclaved soil. This experimental design was a 2 x 2 x 14 factorial consisting of field or autoclaved soil, presence or absence of R. reniformis, and the 12 fungal treatments plus two controls. In both tests, Fusarium oxysporum f. sp. vasinfectum, F. solani, R. solani, and T. basicola consistently displayed extensive root and hypocotyl necrosis that was more severe (P </= 0.05) in the presence of R. reniformis. Soil treatment (autoclaved vs. non-autoclaved) influenced the impact of the Fusarium species on cotton seedling disease, with disease being more severe in the autoclaved soil. Rotylenchulus reniformis reproduction on cotton seedlings was greater in field soil compared to autoclaved soil (P </= 0.05). This study suggests the importance of Fusarium species and R. reniformis in cotton seedling disease.


Plasma FGF23 and Calcified Atherosclerotic Plaque in African Americans with Type 2 Diabetes Mellitus.
Fibroblast growth factor 23 (FGF23) is a phosphaturic hormone implicated in disorders of serum phosphorus concentration and vitamin D. The role of FGF23 in vascular calcification remains controversial. Relationships between FGF23 and coronary artery calcified atherosclerotic plaque (CAC), aortoiliac calcified plaque (CP), carotid artery CP, volumetric bone mineral density (vBMD), albuminuria, and estimated glomerular filtration rate (eGFR) were determined in 545 African Americans with type 2 diabetes (T2D) and preserved kidney function in African American-Diabetes Heart Study participants. Generalized linear models were fitted to test associations between FGF23 and cardiovascular, bone, and renal phenotypes, and change in measurements over time, adjusting for age, gender, African ancestry proportion, body mass index, diabetes duration, hemoglobin A1c, blood pressure, renin-angiotensin-system inhibitors, statins, calcium supplements, serum calcium, and serum phosphate. The sample was 56.7% female with a mean (SD) age of 55.6 (9.6) years, diabetes duration of 10.3 (8.2) years, eGFR 90.9 (22.1) ml/min/1.73 m2, urine albumin:creatinine ratio (UACR) 151 (588) (median 13) mg/g, plasma FGF23 161 (157) RU/ml, and CAC 637 (1,179) mg. In fully adjusted models, FGF23 was negatively associated with eGFR (p < 0.0001) and positively associated with UACR (p < 0.0001) and CAC (p = 0.0006), but not with carotid CP or aortic CP. Baseline FGF23 concentration did not associate with changes in vBMD or CAC after a mean of 5.1 years follow-up. Plasma FGF23 concentrations were independently associated with subclinical coronary artery disease, albuminuria, and kidney function in the understudied African American population with T2D. Findings support relationships between FGF23 and vascular calcification, but not between FGF23 and bone mineral density, in African Americans lacking advanced nephropathy.


miRNA- and cytokine-associated extracellular vesicles mediate squamous cell carcinomas.
Exosomes, or small extracellular vesicles (sEVs), serve as intercellular messengers with key roles in normal and pathological processes. Our previous work had demonstrated that Dsg2 expression in squamous cell carcinoma (SCC) cells enhanced both sEV secretion and loading of pro-mitogenic cargo. In this study, using wild-type Dsg2 and a mutant form that is unable to be palmitoylated (Dsg2cacs), we investigated the mechanism by which Dsg2 modulates SCC tumour development and progression through sEVs. We demonstrate that palmitoylation was required for Dsg2 to regulate sub-cellular localisation of lipid raft and endosomal proteins necessary for sEV biogenesis. Pharmacological inhibition of the endosomal pathway abrogated Dsg2-mediated sEV release. In murine xenograft models, Dsg2-expressing cells generated larger xenograft tumours as compared to cells expressing GFP or Dsg2cacs. Co-treatment with sEVs derived from Dsg2-over-expressing cells increased xenograft size. Cytokine profiling revealed, Dsg2 enhanced both soluble and sEV-associated IL-8 and miRNA profiling revealed, Dsg2 down-regulated both cellular and sEV-loaded miR-146a. miR-146a targets IRAK1, a serine-threonine kinase involved in IL-8 signalling. Treatment with a miR-146a inhibitor up-regulated both IRAK1 and IL-8 expression. RNAseq analysis of HNSCC tumours revealed a correlation between Dsg2 and IL-8. Finally, elevated IL-8 plasma levels were detected in a subset of HNSCC patients who did not respond to immune checkpoint therapy, suggesting that these patients may benefit from prior anti-IL-8 treatment. In summary, these results suggest that intercellular communication through cell-cell adhesion, cytokine release and secretion of EVs are coordinated, and critical for tumour growth and development, and may serve as potential prognostic markers to inform treatment options. Basal cell carcinomas, BCC; Betacellulin, BTC; 2-bromopalmitate, 2-Bromo; Cluster of differentiation, CD; Cytochrome c oxidase IV, COX IV; Desmoglein 2, Dsg2; Early endosome antigen 1, EEA1; Epidermal growth factor receptor substrate 15, EPS15; Extracellular vesicle, EV; Flotillin 1, Flot1; Glyceraldehyde-3-phosphate dehydrogenase, GAPH; Green fluorescent protein, GFP; Head and neck squamous cell carcinoma, HNSCC; Interleukin-1 receptor-associated kinase 1, IRAK1; Interleukin 8, IL-8; Large EV, lEV; MicroRNA, miR; Palmitoylacyltransferase, PAT; Ras-related protein 7 Rab7; Small EV, sEV; Squamous cell carcinoma, SCC; Tissue inhibitor of metalloproteinases, TIMP; Tumour microenvironment, TME.


Core drilling through the ross ice shelf (antarctica) confirmed Basal freezing.
New techniques that have been used to obtain a continuous ice core through the whole 416-meter thickness of the Ross Ice Shelf at Camp J-9 have demonstrated that the bottom 6 meters of the ice shelf consists of sea ice. The rate of basal freezing that is forming this ice is estimated by different methods to be 2 centimeters of ice per year. The sea ice is composed of large vertical crystals, which form the waffle-like lower boundary of the shelf. A distinct alignment of the crystals throughout the sea ice layer suggests the presence of persistent long-term currents beneath the ice shelf.


The effect of ketoconazole on hepatic oxidative drug metabolism in the rat in vivo and in vitro.
Ketoconazole is a new antifungal drug which has increasing clinical application due to its wide spectrum of activity and oral availability. Since substituted imidazoles may be potent inhibitors of cytochrome-mediated drug oxidation, we have examined the effect of ketoconazole on aminopyrine and caffeine metabolism in the rat. The 14C breath test was used to determine the elimination rate of aminopyrine and caffeine in vivo after single and chronic (7 day) treatment with ketoconazole. Acutely, ketoconazole (50 mg/kg) impaired markedly both aminopyrine (56% inhibition) and caffeine demethylation (83% inhibition). Inhibition of aminopyrine and caffeine demethylation as studied with the breath tests was related to ketoconazole dose. The decreased rate of aminopyrine demethylation measured by breath test correlated with decreased aminopyrine clearance from plasma after iv dosage. Chronic treatment with ketoconazole further increased the inhibition of aminopyrine demethylation while chronic treatment was associated with less inhibition of caffeine demethylation than found with a single dose. Breath tests repeated up to 72 hr after ketoconazole treatment revealed differences in aminopyrine and caffeine demethylation. At 24 hr after ketoconazole, aminopyrine demethylation was still inhibited at 78% of controls whereas caffeine demethylation was enhanced to 146% of control. Binding of ketoconazole to microsomal P-450 was determined by spectral analysis. A type II difference spectrum was found with absorbance maximum at 430 nm and minimum at 390 nm. Scatchard analysis revealed a biphasic interaction with estimated dissociation constants of 0.6 and 3.69 microM. Aminopyrine N-demethylation in vitro was markedly impaired with an I50 for ketoconazole of 27 microM and Ki of 8.5 microM.(ABSTRACT TRUNCATED AT 250 WORDS)


Bone marrow stromal cells enhance inter- and intracortical axonal connections after ischemic stroke in adult rats.
We investigated axonal plasticity in the bilateral motor cortices in rats after unilateral stroke and bone marrow stromal cell (BMSC) treatment. Rats were subjected to permanent right middle cerebral artery occlusion followed by intravenous administration of phosphate-buffered saline or BMSCs 1 day later. Adhesive-removal test and modified neurologic severity score were performed weekly to monitor limb functional deficit and recovery. Anterograde tracing with biotinylated dextran amine injected into the right motor cortex was used to assess axonal sprouting in the contralateral motor cortex and ipsilateral rostral forelimb area. Animals were killed 28 days after stroke. Progressive functional recovery was significantly enhanced by BMSCs. Compared with normal animals, axonal density in both contralateral motor cortex and ipsilateral rostral forelimb area significantly increased after stroke. Bone marrow stromal cells markedly enhanced such interhemispheric and intracortical connections. However, labeled transcallosal axons in the corpus callosum were not altered with either stroke or treatment. Both interhemispheric and intracortical axonal sprouting were significantly and highly correlated with behavioral outcome after stroke. This study suggests that, after stroke, cortical neurons surviving in the peri-infarct motor cortex undergo axonal sprouting to restore connections between different cerebral areas. Bone marrow stromal cells enhance axonal plasticity, which may underlie neurologic functional improvement.


Aryl amination using ligand-free Ni(II) salts and photoredox catalysis.
Over the past two decades, there have been major developments in transition metal-catalyzed aminations of aryl halides to form anilines, a common structure found in drug agents, natural product isolates, and fine chemicals. Many of these approaches have enabled highly efficient and selective coupling through the design of specialized ligands, which facilitate reductive elimination from a destabilized metal center. We postulated that a general and complementary method for carbon-nitrogen bond formation could be developed through the destabilization of a metal amido complex via photoredox catalysis, thus providing an alternative approach to the use of structurally complex ligand systems. Here, we report the development of a distinct mechanistic paradigm for aryl amination using ligand-free nickel(II) salts, in which facile reductive elimination from the nickel metal center is induced via a photoredox-catalyzed electron-transfer event.


Whole genome analysis of Leptospira licerasiae provides insight into leptospiral evolution and pathogenicity.
The whole genome analysis of two strains of the first intermediately pathogenic leptospiral species to be sequenced (Leptospira licerasiae strains VAR010 and MMD0835) provides insight into their pathogenic potential and deepens our understanding of leptospiral evolution. Comparative analysis of eight leptospiral genomes shows the existence of a core leptospiral genome comprising 1547 genes and 452 conserved genes restricted to infectious species (including L. licerasiae) that are likely to be pathogenicity-related. Comparisons of the functional content of the genomes suggests that L. licerasiae retains several proteins related to nitrogen, amino acid and carbohydrate metabolism which might help to explain why these Leptospira grow well in artificial media compared with pathogenic species. L. licerasiae strains VAR010(T) and MMD0835 possess two prophage elements. While one element is circular and shares homology with LE1 of L. biflexa, the second is cryptic and homologous to a previously identified but unnamed region in L. interrogans serovars Copenhageni and Lai. We also report a unique O-antigen locus in L. licerasiae comprised of a 6-gene cluster that is unexpectedly short compared with L. interrogans in which analogous regions may include >90 such genes. Sequence homology searches suggest that these genes were acquired by lateral gene transfer (LGT). Furthermore, seven putative genomic islands ranging in size from 5 to 36 kb are present also suggestive of antecedent LGT. How Leptospira become naturally competent remains to be determined, but considering the phylogenetic origins of the genes comprising the O-antigen cluster and other putative laterally transferred genes, L. licerasiae must be able to exchange genetic material with non-invasive environmental bacteria. The data presented here demonstrate that L. licerasiae is genetically more closely related to pathogenic than to saprophytic Leptospira and provide insight into the genomic bases for its infectiousness and its unique antigenic characteristics.


Autophagy orchestrates the regulatory program of tumor-associated myeloid-derived suppressor cells.
Myeloid-derived suppressor cells (MDSCs) densely accumulate into tumors and potently suppress antitumor immune responses, promoting tumor development. Targeting MDSCs in tumor immunotherapy has been hampered by lack of understanding of the molecular pathways that govern MDSC differentiation and function. Herein, we identify autophagy as a crucial pathway for MDSC-mediated suppression of antitumor immunity. Specifically, MDSCs in patients with melanoma and mouse melanoma exhibited increased levels of functional autophagy. Ablation of autophagy in myeloid cells markedly delayed tumor growth and endowed antitumor immune responses. Notably, tumor-infiltrating autophagy-deficient monocytic MDSCs (M-MDSCs) demonstrated impaired suppressive activity in vitro and in vivo, whereas transcriptome analysis revealed substantial differences in genes related to lysosomal function. Accordingly, autophagy-deficient M-MDSCs exhibited impaired lysosomal degradation, thereby enhancing surface expression of MHC class II molecules, resulting in efficient activation of tumor-specific CD4+ T cells. Finally, targeting of the membrane-associated RING-CH1 (MARCH1) E3 ubiquitin ligase that mediates the lysosomal degradation of MHC II in M-MDSCs attenuated their suppressive function, and resulted in markedly decreased tumor volume followed by development of a robust antitumor immunity. Collectively, these findings depict autophagy as a molecular target of MDSC-mediated suppression of antitumor immunity.


β-adrenergic receptor mediation of stress-induced reinstatement of extinguished cocaine-induced conditioned place preference in mice: roles for β1 and β2 adrenergic receptors.
Stress can trigger the relapse of drug use in recovering cocaine addicts and reinstatement in rodent models through mechanisms that may involve norepinephrine release and β-adrenergic receptor activation. The present study examined the role of β-adrenergic receptor subtypes in the stressor-induced reinstatement of extinguished cocaine-induced (15 mg/kg i.p.) conditioned place preference in mice. Forced swim (6 min at 22°C) stress or activation of central noradrenergic neurotransmission by administration of the selective α(2) adrenergic receptor antagonist 2-[(4,5-dihydro-1H-imidazol-2-yl)methyl]-2,3-dihydro-1-methyl-1H-isoindole (BRL-44,408) (10 mg/kg i.p.) induced reinstatement in wild-type, but not β- adrenergic receptor-deficient Adrb1/Adrb2 double-knockout, mice. In contrast, cocaine administration (15 mg/kg i.p.) resulted in reinstatement in both wild-type and β-adrenergic receptor knockout mice. Stress-induced reinstatement probably involved β(2) adrenergic receptors. The β(2) adrenergic receptor antagonist -(isopropylamino)-1-[(7-methyl-4-indanyl)oxy]butan-2-ol (ICI-118,551) (1 or 2 mg/kg i.p.) blocked reinstatement by forced swim or BRL-44,408, whereas administration of the nonselective β-adrenergic receptor agonist isoproterenol (2 or 4 mg/kg i.p.) or the β(2) adrenergic receptor-selective agonist clenbuterol (2 or 4 mg/kg i.p.) induced reinstatement. Forced swim-induced, but not BRL-44,408-induced, reinstatement was also blocked by a high (20 mg/kg) but not low (10 mg/kg) dose of the β(1) adrenergic receptor antagonist betaxolol, and isoproterenol-induced reinstatement was blocked by pretreatment with either ICI-118,551 or betaxolol, suggesting a potential cooperative role for β(1) and β(2) adrenergic receptors in stress-induced reinstatement. Overall, these findings suggest that targeting β-adrenergic receptors may represent a promising pharmacotherapeutic strategy for preventing drug relapse, particularly in cocaine addicts whose drug use is stress related.


The new Avicenna Roboflex: How does the irrigation system work? Results from an in vitro experiment.
Since 2012 Elmed has been working on a robot specifically designed for flexible ureteroscopy. After the first version of Avicenna Roboflex, a second version was developed in 2015, with significant changes especially in the irrigation system. We consider mandatory for the endourologist that works with the Avicenna Roboflex be aware of the functioning of the irrigation system. We connected a container to the pump's irrigation system and measured the quantity of saline per second delivered by each speed setting, with/without the flush in five different modalities: pump on its own, pump with ureteroscope, with two laser fibers, with 1.9 Fr basket, and with a Terumo guidewire. The highest mean flow-rates were observed in the 200- micrometer laser fiber, after the pump on its own. Median flowrates for all speed settings were significantly higher for the pump on its own than for the URS in both flushed and nonflushed modes (p = 0.045, p = 0.039 respectively). There was no statistically significant difference in median flow-rates between the guide wire and basket in all of the speed settings (p = 0.932 and p = 0.977). For both laser fibers there was no statistically significant difference between the median flow rate on both nonflush and flush modes. (p = 0.590 & p = 0.590). There was a linear correlation between the speed setting and the increase measured with the flush-option for pump only measurements (r = 0.602, p = 0.038). There was no statistically significant difference between laser fibers and the pump on its own on the increase of flow rate with flush mode. (p = 0.443 for the 272- micrometer fiber and p = 0.219 for the 200-micrometer fiber). The irrigation system of the new Avicenna Roboflex is optimized compared to the previous version. However other more complex studies concerning the live flow/pressure relationship are needed before firm conclusions can be made.


Maternal fish consumption, hair mercury, and infant cognition in a U.S. Cohort.
Fish and other seafood may contain organic mercury but also beneficial nutrients such as n-3 polyunsaturated fatty acids. We endeavored to study whether maternal fish consumption during pregnancy harms or benefits fetal brain development. We examined associations of maternal fish intake during pregnancy and maternal hair mercury at delivery with infant cognition among 135 mother-infant pairs in Project Viva, a prospective U.S. pregnancy and child cohort study. We assessed infant cognition by the percent novelty preference on visual recognition memory (VRM) testing at 6 months of age. Mothers consumed an average of 1.2 fish servings per week during the second trimester. Mean maternal hair mercury was 0.55 ppm, with 10% of samples > 1.2 ppm. Mean VRM score was 59.8 (range, 10.9-92.5). After adjusting for participant characteristics using linear regression, higher fish intake was associated with higher infant cognition. This association strengthened after adjustment for hair mercury level: For each additional weekly fish serving, offspring VRM score was 4.0 points higher [95% confidence interval (CI), 1.3 to 6.7]. However, an increase of 1 ppm in mercury was associated with a decrement in VRM score of 7.5 (95% CI, -13.7 to -1.2) points. VRM scores were highest among infants of women who consumed > 2 weekly fish servings but had mercury levels </= 1.2 ppm. Higher fish consumption in pregnancy was associated with better infant cognition, but higher mercury levels were associated with lower cognition. Women should continue to eat fish during pregnancy but choose varieties with lower mercury contamination.


Massage therapy usage and reported health in older adults experiencing persistent pain.
Persistent pain is a frequent complaint among older adults and can greatly decrease quality of life while also contributing to other negative outcomes such as poor health, increased pharmaceutical medication usage, increased rates of depression, and cognitive decline. The current study (N = 69) examines the potential impact of massage therapy (MT) in older adults (60+ years) with persistent pain, by comparing self-reported health outcome scores among those who have and have not utilized massage therapy in the past year. The current study was derived from a larger study that collected data as part of a one-time, self-report, mail-in survey. Lexington, Kentucky area adults, 60 and older who reported persistent pain were eligible to participate in the study. The RAND 36-Item Health Survey was used to determine participant health-related quality of life. The current study demonstrated that for older adults experiencing persistent pain, massage is associated with self-report of less limitation due to physical or emotional issues, better emotional health, more energy/less fatigue, better social functioning, and better overall health. Age, education, cumulative morbidities, number of areas in which participants reported experiencing persistent pain, and number of complementary and alternative medicine options in addition to MT utilized in the past year did not affect the association between receipt of massage and better self-reports in those domains. While many causes of pain for older adults elude cure, further study is warranted that examines MT as an intervention to improve coping in older adults with persistent pain.


Targeting green fluorescent protein to dendritic membrane in central neurons.
Dendritic and axonal processes are input and output sites, respectively, of neuronal information, and detailed visualization of these processes may be indispensable for elucidating the neuronal circuits and revealing the principles of neuronal functions. To establish a method for completely visualizing dendritic processes, we first developed green fluorescent protein (GFP)-based proteins and, by using lentivirus with a neuron-specific promoter, examined whether or not the protein fully visualized the dendritic processes of infected neurons. When GFP with a palmitoylation (palGFP) or myristoylation/palmitoylation site (myrGFP) was expressed in rat brain with lentiviruses, myrGFP labeled dendritic membrane better than palGFP. Subsequently, dendrite-targeting efficiencies of three basolateral membrane-sorting and three putative dendrite-targeting domains, which were attached to myrGFP C-terminus, were examined in striatonigral GABAergic and corticothalamic glutamatergic neurons, and in cultured cortical neurons. Of the six domains, C-terminal cytoplasmic domain of low density lipoprotein receptor (LDLRct) was most efficient in targeting the protein to dendrites, showing 8.5-15-fold higher efficiency in striatonigral neurons compared with myrGFP. Finally, dendritic membrane-targeting potency of myrGFP-LDLRct was confirmed in transgenic mice using Thy1 or Gad1 expression cassette. Thus, myrGFP-LDLRct is an excellent synthetic protein for dendritic visualization, and may be a useful tool for the morphological analysis of neuronal circuits.


Epidermal Langerhans' cell induction of immunity against an ultraviolet-induced skin tumour.
Langerhans' cells (LC) have been shown experimentally to induce immune responses against many antigens; however, their role in the initiation of anti-tumour immunity has received little attention. This study examined the ability of murine epidermal LC to induce immunity to an ultraviolet radiation (UV)-induced skin tumour. Freshly prepared epidermal cells (EC) were cultured for 2 or 20 hr with granulocyte-macrophage colony-stimulating factor (GM-CSF), pulsed with an extract of the UV-13-1 tumour, then used to immunize naive syngeneic mice. Delayed type hypersensitivity (DTH) was elicited 10 days after immunization by injection of UV-13-1 tumour cells into the ear pinna, and measured 24 hr later. EC cultured with GM-CSF for 2 hr induced antitumour DTH, as did EC cultured for 20 hr without GM-CSF. Conversely, EC cultured for 2 hr without GM-CSF, or EC cultured for 20 hr with GM-CSF were unable to induce a DTH. Induction of immunity required active presentation of tumour antigens by Ia+ EC and was tumour specific. Thus Ia+ epidermal cells are capable of inducing anti-tumour immunity to UV-induced skin tumours, but only when they contact antigen in particular states of maturation.


The influence of coastal access on isotope variation in Icelandic arctic foxes.
To quantify the ecological effects of predator populations, it is important to evaluate how population-level specializations are dictated by intra- versus inter-individual dietary variation. Coastal habitats contain prey from the terrestrial biome, the marine biome and prey confined to the coastal region. Such habitats have therefore been suggested to better support predator populations compared to habitats without coastal access. We used stable isotope data on a small generalist predator, the arctic fox, to infer dietary strategies between adult and juvenile individuals with and without coastal access on Iceland. Our results suggest that foxes in coastal habitats exhibited a broader isotope niche breadth compared to foxes in inland habitats. This broader niche was related to a greater diversity of individual strategies rather than to a uniform increase in individual niche breadth or by individuals retaining their specialization but increasing their niche differentiation. Juveniles in coastal habitats exhibited a narrower isotope niche breadth compared to both adults and juveniles in inland habitats, and juveniles in inland habitats inhabited a lower proportion of their total isotope niche compared to adults and juveniles from coastal habitats. Juveniles in both habitats exhibited lower intra-individual variation compared to adults. Based on these results, we suggest that foxes in both habitats were highly selective with respect to the resources they used to feed offspring, but that foxes in coastal habitats preferentially utilized marine resources for this purpose. We stress that coastal habitats should be regarded as high priority areas for conservation of generalist predators as they appear to offer a wide variety of dietary options that allow for greater flexibility in dietary strategies.


Analytical validation of the Oncotype DX prostate cancer assay - a clinical RT-PCR assay optimized for prostate needle biopsies.
The Oncotype DX Prostate Cancer Assay is a multi-gene RT-PCR expression assay that was developed for use with fixed paraffin-embedded (FPE) diagnostic prostate needle biopsies containing as little as 1 mm of prostate tumor in the greatest dimension. The assay measures expression of 12 cancer-related genes representing four biological pathways and 5 reference genes which are algorithmically combined to calculate the Genomic Prostate Score (GPS). This biopsy-based assay has been analytically and subsequently clinically validated as a predictor of aggressive prostate cancer. The aim of this study was to validate the analytical performance of the Oncotype DX Prostate Cancer Assay using predefined acceptance criteria. The lowest quartile of RNA yields from prostate needle biopsies (six 5 μm sections) was between 19 and 34 ng. Analytical validation of the process requiring as little as 5 ng of RNA met all pre-defined acceptance criteria. Amplification efficiencies, analytical sensitivity, and accuracy of gene assays were measured by serially diluting an RNA sample and analyzing features of the linear regression between RNA expression measured by the crossing point (Cp) versus the log2 of the RNA input per PCR assay well. Gene assays were shown to accurately measure expression over a wide range of inputs (from as low as 0.005 ng to 320 ng). Analytical accuracy was excellent with average biases at qPCR inputs representative of patient samples <9.7% across all assays while amplification efficiencies were within ±6% of the median. Assessments of reproducibility and precision were performed by testing 10 prostate cancer RNA samples over multiple instruments, reagent lots, operators, days (precision), and RNA input levels (reproducibility) using appropriately parameterized linear mixed models. The standard deviations for analytical precision and reproducibility were 1.86 and 2.11 GPS units (100-unit scale) respectively. The Oncotype DX Prostate Cancer Assay, a clinical RT-PCR assay specifically designed for use with prostate needle biopsies, has been analytically validated using very limited RNA inputs. The assay requirements and analytical performance will provide physicians with test results from a robust and reliable assay which will enable improved treatment decisions for men diagnosed with early-stage prostate cancer.


A particular effect of sleep, but not pain or depression, on the blood-oxygen-level dependent response during working memory tasks in patients with chronic pain.
Patients with chronic pain (CP) are often reported to have deficits in working memory. Pain impairs working memory, but so do depression and sleep problems, which are also common in CP. Depression has been linked to changes in brain activity in CP during working memory tasks, but the effect of sleep problems on working memory performance and brain activity remains to be investigated. Fifteen CP patients and 17 age-, sex-, and education-matched controls underwent blood-oxygen-level dependent (BOLD) functional magnetic resonance imaging at 3T while performing block design 0-back, 2-back, and paced visual serial addition test paradigms. Subjects also reported their level of pain (Brief Pain Inventory), depression (Beck Depression Inventory II), and sleep problems (Pittsburgh Sleep Quality Index) and were tested outside the scanner with neuropsychological tests of working memory. The CP group reported significantly higher levels of pain, depression, and sleep problems. No significant performance difference was found on the neuropsychological tests in or outside the scanner between the two groups. There were no correlations between level of pain, depression, and sleep problems or between these and the neuropsychological test scores. CP patients exhibited significantly less brain activation and deactivation than controls in parietal and frontal lobes, which are the brain areas that normally show activation and deactivation during working memory tasks. Sleep problems independently and significantly modulated the BOLD response to the complex working memory tasks and were associated with decreased brain activation in task-positive regions and decreased deactivation in the default mode network in the CP group compared to the control group. The pain and depression scores covaried with working memory activation. Sleep problems in CP patients had a significant impact on the BOLD response during working memory tasks, independent of pain level and depression, even when performance was shown not to be significantly affected.


Altered cytokine export and apoptosis in mice deficient in interleukin-1 beta converting enzyme.
The interleukin-1 beta (IL-1 beta) converting enzyme (ICE) processes the inactive IL-1 beta precursor to the proinflammatory cytokine. Adherent monocytes from mice harboring a disrupted ICE gene (ICE-/-) did not export IL-1 beta or interleukin-1 alpha (IL-1 alpha) after stimulation with lipopolysaccharide. Export of tumor necrosis factor-alpha and interleukin-6 (IL-6) from these cells was also diminished. Thymocytes from ICE-/- mice were sensitive to apoptosis induced by dexamethasone or ionizing radiation, but were resistant to apoptosis induced by Fas antibody. Despite this defect in apoptosis, ICE-/- mice proceed normally through development.


Multiscale Assembly of Grape-Like Ferroferric Oxide and Carbon Nanotubes: A Smart Absorber Prototype Varying Temperature to Tune Intensities.
Ideal electromagnetic attenuation material should not only shield the electromagnetic interference but also need strong absorption. Lightweight microwave absorber with thermal stability and high efficiency is a highly sought-after goal of researchers. Tuning microwave absorption to meet the harsh requirements of thermal environments has been a great challenge. Here, grape-like Fe3O4-multiwalled carbon nanotubes (MWCNTs) are synthesized, which have unique multiscale-assembled morphology, relatively uniform size, good crystallinity, high magnetization, and favorable superparamagnetism. The Fe3O4-MWCNTs is proven to be a smart microwave-absorber prototype with tunable high intensities in double belts in the temperature range of 323-473 K and X band. Maximum absorption in two absorbing belts can be simultaneously tuned from ∼-10 to ∼-15 dB and from ∼-16 to ∼-25 dB by varying temperature, respectively. The belt for reflection loss ≤-20 dB can almost cover the X band at 323 K. The tunable microwave absorption is attributed to effective impedance matching, benefiting from abundant interfacial polarizations and increased magnetic loss resulting from the grape-like Fe3O4 nanocrystals. Temperature adjusts the impedance matching by changing both the dielectric and magnetic loss. The special assembly of MWCNTs and magnetic loss nanocrystals provides an effective pathway to realize excellent absorbers at elevated temperature.


3D Mass Spectrometry Imaging Reveals a Very Heterogeneous Drug Distribution in Tumors.
Mass Spectrometry Imaging (MSI) is a widespread technique used to qualitatively describe in two dimensions the distribution of endogenous or exogenous compounds within tissue sections. Absolute quantification of drugs using MSI is a recent challenge that just in the last years has started to be addressed. Starting from a two dimensional MSI protocol, we developed a three-dimensional pipeline to study drug penetration in tumors and to develop a new drug quantification method by MALDI MSI. Paclitaxel distribution and concentration in different tumors were measured in a 3D model of Malignant Pleural Mesothelioma (MPM), which is known to be a very heterogeneous neoplasm, highly resistant to different drugs. The 3D computational reconstruction allows an accurate description of tumor PTX penetration, adding information about the heterogeneity of tumor drug distribution due to the complex microenvironment. The use of an internal standard, homogenously sprayed on tissue slices, ensures quantitative results that are similar to those obtained using HPLC. The 3D model gives important information about the drug concentration in different tumor sub-volumes and shows that the great part of each tumor is not reached by the drug, suggesting the concept of pseudo-resistance as a further explanation for ineffective therapies and tumors relapse.


[Interstitial percutaneous therapies in primary and secondary liver tumors].
Multiple percutaneous interstitial systems to induce intrahepatic tumor necrosis are classifiable in two major groups: those using chemical agents (ethanol and acetic acid) and those adopting thermal effects (hot saline, radiofrequency, laser and microwave). Indication, technique and results of percutaneous ethanol injection (PEI) are considered. In the case of single hepatocellular carcinoma (HCC), < 5 cm and associated with cirrhosis Child class A long-term results of PEI are comparable with those of best surgical series with 5-years survival which range from 47 to 51%. No mortality and recurrence rates similar to those of surgical cases make PEI as an alternative to surgical resection although the difficulties in comparing retrospectively the different experiences do not enable definitive conclusions. In the case of liver metastases from colorectal cancer or endocrine tumors, which seem more sensitive to locoregional therapies, complete response rates are not adequate yet being 86%, 26%, 11% and 0% respectively in the case of < 2 cm, 2-3 cm, 3-4 cm and > 4 cm lesions and moreover no definitive data are available about long-term prognosis. Waiting for further validation about the encouraging data obtained with the other interstitial methods and in particular with radiofrequency (RF) and laser interstitial photocoagulation (FLI), PEI is a valid alternative to surgery for selected HCC patients and is a good palliation in cases with not resectable liver metastases which are anyway suitable for locoregional treatments.


Undiagnosed bipolar disorder in patients treated for major depression in China.
Bipolar disorder (BD) is a recurrent, complex illness and often misdiagnosed and treated as a major depressive disorder (MDD). This study set out (1) to investigate the proportion of BD in patients treated for MDD using DSM-IV diagnostic criteria; (2) to test the usefulness of the screening tool - the 32-item Hypomania Checklist (HCL-32) in Chinese patients; and (3) to assess whether MDD patients with subthreshold manic features (patients who screened positive for BD on the HCL-32, but did not meet the diagnostic criteria for DSM-IV BD as measured by Mini International Neuropsychiatric Interview (MINI)) differ from those with BD, and from those suffering from MDD without manic features in terms of basic demographic and clinical variables. A total of 1487 patients treated for MDD were consecutively examined in 13 mental health centers in China. The patients' socio-demographic and clinical characteristics were recorded using a standardized protocol and data collection procedure. The HCL-32 was self-completed by patients to identify hypomanic symptoms, and the MINI was used by clinicians to establish DSM-IV diagnoses. The proportions of undiagnosed BD (all types), BD-I and BD-II were 20.8%, 7.9% and 12.8%, respectively. The HCL-32 had low positive predictive value (0.43). Compared to MDD patients without subthreshold manic features, MDD patients with subthreshold manic features were younger at onset, less likely to be married and had more depressive episodes on a seasonal basis, and more frequent depressive episodes overall. Compared to BD patients, MDD patients with subthreshold manic features had an older age at onset and less frequent depressive episodes and less family history of psychiatric disorders, appetite, weight gain and time spent sleeping, suicide ideation and attempts and psychotic symptoms. At least one fifth of Chinese patients treated for MDD may have an undiagnosed BD. The HCL-32 is useful to identify broader subthreshold bipolar features. The findings need to be confirmed by longitudinal studies using more comprehensive, standardized instruments.


Coding of hemolysins within the ribosomal RNA repeat on a plasmid in Entamoeba histolytica.
The pathogenesis of amoebic dysentery is a result of cytolysis of the colonic mucosa by the parasitic protozoan Entamoeba histolytica. The cytolysis results in extensive local ulceration and allows the amoeba to penetrate and metastasize to distant sites. Factors involved in this process were defined with three clones that express hemolytic activities in Escherichia coli. These potential amoebic virulence determinants were also toxic to human colonic epithelial cells, the primary cellular targets in amoebal invasion of the large intestine. The coding sequences for the hemolysins were close to each other on a 2.6-kilobase segment of a 25-kilobase extrachromosomal DNA element. The structural genes for the hemolysins were within inverted repeats that encode ribosomal RNAs.


The Imaginative Work of Tim Hengst.
This JBC Showcase features the creative and imaginative work of Timothy Hengst, who was the recipient of the Association of Medical Illustrators' Lifetime Achievement Award in 2013. This Award is the highest honor awarded by the Association of Medical Illustrators to an individual who has dedicated his or her professional life as a medical illustrator. The JBC is proud to present these medical illustrations that represent Mr. Hengst's life's work.


Quantifying lipid contents in enveloped virus particles with plasmonic nanoparticles.
Phosphatidylserine (PS) and monosialotetrahexosylganglioside (GM1 ) are examples of two host-derived lipids in the membrane of enveloped virus particles that are known to contribute to virus attachment, uptake, and ultimately dissemination. A quantitative characterization of their contribution to the functionality of the virus requires information about their relative concentrations in the viral membrane. Here, a gold nanoparticle (NP) binding assay for probing relative PS and GM1 lipid concentrations in the outer leaflet of different HIV-1 and Ebola virus-like particles (VLPs) using sample sizes of less than 3 × 10(6) particles is introduced. The assay evaluates both scattering intensity and resonance wavelength, and determines relative NP densities through plasmon coupling as a measure for the target lipid concentrations in the NP-labeled VLP membrane. A correlation of the optical observables with absolute lipid contents is achieved by calibration of the plasmon coupling-based methodology with unilamellar liposomes of known PS or GM1 concentration. The performed studies reveal significant differences in the membrane of VLPs that assemble at different intracellular sites and pave the way to an optical quantification of lipid concentration in virus particles at physiological titers.


Polymorphisms in innate immunity genes and risk of childhood leukemia.
The immune system plays an important role in the control of cancer development. To investigate possible genetic contribution to childhood leukemia risk in the innate immune system, we performed an association study for the 1214 SNPs in 146 gene regions related to innate immunity using GoldenGate (Illumina) oligonucleotide pool assay (OPA) in 106 case patients and 123 controls. Childhood leukemia risk was estimated as odds ratios and 95% confidence intervals adjusted for age, gender and birth weight. The minP test was used to identify statistically significant association at gene level. Three SNPs (STAT6 rs703817, C1qG rs17433222, and MBP rs3794845) were significantly associated with childhood leukemia risk (p(trend) < 0.001, minP < 0.01). The most significant association with childhood leukemia risk was for STAT6 rs703817 (GA vs GG: 0.48 (0.26-0.87), AA vs GG: 0.21 (0.07-0.61), p(trend) = 0.0003, minP = 0.002). Subgroup analysis showed that Ly96 rs78380171 and MBP rs3794845 were significantly associated with the risk of acute lymphoblastic leukemia (p(trend) < 0.001). Our results suggest that genetic polymorphisms in innate immunity genes might play a role in the genesis of childhood leukemia with limited biologic evidence. Additional, larger studies are needed to identify the mechanism of these genes in childhood leukemia patients.


Variations in cataract extraction rates in Medicare prepaid and fee-for-service settings.
To compare rates of cataract extraction in 2 prepaid health settings and in traditional fee-for-service (FFS) settings. A cross-sectional analysis using 1993 health maintenance organization (HMO) Medicare claims and encounter files, the Health Care Financing Administration (HCFA) 5% Medicare Part B provider/supplier file, and the HCFA October 1992 100% Medicare population file. Southern California Medicare FFS settings and the staff-model and independent practice association (IPA) plans of a large California HMO. 1993 Medicare beneficiaries aged 65 years and older. The study included 43387 staff-model HMO enrollees, 19050 IPA enrollees, and 47 150 FFS beneficiaries (a 5% sample of all Southern California FFS beneficiaries). Age and risk-factor adjusted rates of cataract extraction per 1000 beneficiary-years. After controlling for age, sex, and diabetes mellitus status, FFS beneficiaries were twice as likely to undergo cataract extraction as were prepaid beneficiaries (P<.01). Female FFS beneficiaries were nearly twice as likely to undergo the procedure as were male FFS beneficiaries (P<.001); there were no extraction rate differences by sex in the prepaid settings. Because of the potential implications for vision care in the elderly, the significantly different rates of cataract extraction in FFS and prepaid settings warrant further clinical investigation to determine whether there is overuse in FFS vs underuse in prepaid settings. Such investigations must assess the appropriateness of cataract surgery by evaluating its use relative to clinical need.


PD-1 Blockade with Pembrolizumab in Advanced Merkel-Cell Carcinoma.
Merkel-cell carcinoma is an aggressive skin cancer that is linked to exposure to ultraviolet light and the Merkel-cell polyomavirus (MCPyV). Advanced Merkel-cell carcinoma often responds to chemotherapy, but responses are transient. Blocking the programmed death 1 (PD-1) immune inhibitory pathway is of interest, because these tumors often express PD-L1, and MCPyV-specific T cells express PD-1. In this multicenter, phase 2, noncontrolled study, we assigned adults with advanced Merkel-cell carcinoma who had received no previous systemic therapy to receive pembrolizumab (anti-PD-1) at a dose of 2 mg per kilogram of body weight every 3 weeks. The primary end point was the objective response rate according to Response Evaluation Criteria in Solid Tumors, version 1.1. Efficacy was correlated with tumor viral status, as assessed by serologic and immunohistochemical testing. A total of 26 patients received at least one dose of pembrolizumab. The objective response rate among the 25 patients with at least one evaluation during treatment was 56% (95% confidence interval [CI], 35 to 76); 4 patients had a complete response, and 10 had a partial response. With a median follow-up of 33 weeks (range, 7 to 53), relapses occurred in 2 of the 14 patients who had had a response (14%). The response duration ranged from at least 2.2 months to at least 9.7 months. The rate of progression-free survival at 6 months was 67% (95% CI, 49 to 86). A total of 17 of the 26 patients (65%) had virus-positive tumors. The response rate was 62% among patients with MCPyV-positive tumors (10 of 16 patients) and 44% among those with virus-negative tumors (4 of 9 patients). Drug-related grade 3 or 4 adverse events occurred in 15% of the patients. In this study, first-line therapy with pembrolizumab in patients with advanced Merkel-cell carcinoma was associated with an objective response rate of 56%. Responses were observed in patients with virus-positive tumors and those with virus-negative tumors. (Funded by the National Cancer Institute and Merck; ClinicalTrials.gov number, NCT02267603.).


Assessing recovery of in vitro steroid production in male rainbow darter (Etheostoma caeruleum) in response to municipal wastewater treatment plant infrastructure changes.
The present study examined in vitro 11-ketotestosterone and testosterone production by the testes of rainbow darter (Etheostoma caeruleum) collected from selected reference sites and downstream of 2 municipal wastewater treatment plants (MWWTPs; Waterloo and Kitchener) on the central Grand River (Ontario, Canada), over a 6-yr period (2011-2016). The main objective was to investigate if infrastructure upgrades at the Kitchener MWWTP in 2012 resulted in a recovery of this response in the post-upgrade period (2013-2016). Two supporting studies showed that the fall season is appropriate for measuring in vitro sex steroid production because it provides stable detection of steroid patterns, and that the sample handling practiced in the present study did not introduce a bias. Infrastructure upgrades of the Kitchener MWWTP resulted in significant reductions in ammonia and estrogenicity. After the upgrades, 11-ketotestosterone production by MWWTP-exposed fish increased in 2013 and it continued to recover throughout the study period of 2014 through 2016, returning to levels measured in reference fish. Testosterone production was less sensitive and it lacked consistency. The Waterloo MWWTP underwent some minor upgrades but the level of ammonia and estrogenicity remained variable over time. The production of 11-ketotestosterone and testosterone in rainbow darter below the Waterloo MWWTP was variable and without a clear recovery pattern over the course of the present study. The results of the present study demonstrated that measuring production of sex steroids (especially 11-ketotestosterone) over multiple years can be relevant for assessing responses in fish to environmental changes such as those resulting from major infrastructure upgrades. Environ Toxicol Chem 2018;37:501-514. © 2017 SETAC.


HbF Levels in Sickle Cell Disease Are Associated with Proportion of Circulating Hematopoietic Stem and Progenitor Cells and CC-Chemokines.
The concentration of circulating hematopoietic stem and progenitor cells has not been studied longitudinally. Here, we report that the proportions of Lin-CD34+38- hematopoietic multipotent cells (HMCs) and of Lin-CD34+CD38+ hematopoietic progenitors cells (HPCs) are highly variable between individuals but stable over long periods of time, in both healthy individuals and sickle cell disease (SCD) patients. This suggests that these proportions are regulated by genetic polymorphisms or by epigenetic mechanisms. We also report that in SCD patients treated with hydroxyurea, the proportions of circulating HMCs and HPCs show a strong positive and negative correlation with fetal hemoglobin (HbF) levels, respectively. Titration of 65 cytokines revealed that the plasma concentration of chemokines CCL2, CCL11, CCL17, CCL24, CCL27, and PDGF-BB were highly correlated with the proportion of HMCs and HPCs and that a subset of these cytokines were also correlated with HbF levels. A linear model based on four of these chemokines could explain 80% of the variability in the proportion of circulating HMCs between individuals. The proportion of circulating HMCs and HPCs and the concentration of these chemokines might therefore become useful biomarkers for HbF response to HU in SCD patients. Such markers might become increasingly clinically relevant, as alternative treatment modalities for SCD are becoming available.


The Development of a Skin Cancer Classification System for Pigmented Skin Lesions Using Deep Learning.
Recent studies have demonstrated the usefulness of convolutional neural networks (CNNs) to classify images of melanoma, with accuracies comparable to those achieved by dermatologists. However, the performance of a CNN trained with only clinical images of a pigmented skin lesion in a clinical image classification task, in competition with dermatologists, has not been reported to date. In this study, we extracted 5846 clinical images of pigmented skin lesions from 3551 patients. Pigmented skin lesions included malignant tumors (malignant melanoma and basal cell carcinoma) and benign tumors (nevus, seborrhoeic keratosis, senile lentigo, and hematoma/hemangioma). We created the test dataset by randomly selecting 666 patients out of them and picking one image per patient, and created the training dataset by giving bounding-box annotations to the rest of the images (4732 images, 2885 patients). Subsequently, we trained a faster, region-based CNN (FRCNN) with the training dataset and checked the performance of the model on the test dataset. In addition, ten board-certified dermatologists (BCDs) and ten dermatologic trainees (TRNs) took the same tests, and we compared their diagnostic accuracy with FRCNN. For six-class classification, the accuracy of FRCNN was 86.2%, and that of the BCDs and TRNs was 79.5% (p = 0.0081) and 75.1% (p < 0.00001), respectively. For two-class classification (benign or malignant), the accuracy, sensitivity, and specificity were 91.5%, 83.3%, and 94.5% by FRCNN; 86.6%, 86.3%, and 86.6% by BCD; and 85.3%, 83.5%, and 85.9% by TRN, respectively. False positive rates and positive predictive values were 5.5% and 84.7% by FRCNN, 13.4% and 70.5% by BCD, and 14.1% and 68.5% by TRN, respectively. We compared the classification performance of FRCNN with 20 dermatologists. As a result, the classification accuracy of FRCNN was better than that of the dermatologists. In the future, we plan to implement this system in society and have it used by the general public, in order to improve the prognosis of skin cancer.


CrkRS: a novel conserved Cdc2-related protein kinase that colocalises with SC35 speckles.
We have isolated and characterised a novel human protein kinase, Cdc2-related kinase with an arginine/serine-rich (RS) domain (CrkRS), that is most closely related to the cyclin-dependent kinase (CDK) family. CrkRS is a 1490 amino acid protein, the largest CDK-related kinase so far isolated. The protein kinase domain of CrkRS is 89% identical to the 46 kDa CHED protein kinase, but outside the kinase domains the two proteins are completely unrelated. CrkRS has extensive proline-rich regions that match the consensus for SH3 and WW domain binding sites, and an RS domain that is predominantly found in splicing factors. CrkRS is ubiquitously expressed in tissues, and maps to a single genetic locus. There are closely related protein kinases in both the Drosophila and Caenorhabditis elegans genomes. Consistent with the presence of an RS domain, anti-CrkRS antibodies stain nuclei in a speckled pattern, overlapping with spliceosome components and the hyperphosphorylated form of RNA polymerase II. Like RNA polymerase II, CrkRS is a constitutive MPM-2 antigen throughout the cell cycle. Anti-CrkRS immunoprecipitates phosphorylate the C-terminal domain of RNA polymerase II in vitro. Thus CrkRS may be a novel, conserved link between the transcription and splicing machinery.


Low-dose IFN-gamma induces tumor MHC expression in metastatic malignant melanoma.
Specific antitumor immune responses require expression of MHC class I or II molecules on tumor cells, and MHC antigen down-regulation is a presumed tumor growth promoting mechanism. Because IFN-gamma up-regulates tumor MHC antigen expression in vitro, in this Phase II trial of an immunologically active dose and schedule we evaluated whether this was the case in vivo. Twenty-three patients with metastatic melanoma were treated with IFN-gamma 100 microg/m(2) s.c. once weekly for a maximum of 6 months. There were three complete responses, now maintained for 53, 36, and 25 months. The remainder had progressive disease. The treatment was well tolerated, with no toxicity exceeding National Cancer Institute Common Toxicity Criteria grade II. Immunohistochemical analysis of tumor biopsies during treatment was performed using monoclonal antibodies to HLA class I (W/632) and class II (CR3/43) monomorphic determinants. HLA class I was down-regulated in 2 of 19 patients pretreatment and up-regulated by IFN-gamma in both. HLA class II was down-regulated pretreatment in 14 of 18 patients and up-regulated by IFN-gamma in 6 (43%). The HLA up-regulation persisted throughout the study. IFN-gamma induced significant but short-lived up-regulation of surrogate markers of monocyte activation (serum neopterin) and class I up-regulation (serum beta-2-microglobulin) in most patients. There was no consistent relationship between surrogate marker up-regulation, tumor antigen up-regulation, and responses. The study shows that the significant immune modulation induced by IFN-gamma does not correlate with tumor responses and that the serum surrogate marker changes do not reflect tumor events. The durable and long-lived responses, clear demonstration of tumor MHC up-regulation, and low toxicity suggest that weekly IFN-gamma 100 microg/m(2) would be a useful addition to chemoimmunotherapeutic regimens.


Molecular characterisation and origin of the Coffea arabica L. genome.
Restriction fragment length polymorphism (RFLP) markers were used in combination with genomic in situ hybridisation (GISH) to investigate the origin of the allotetraploid species Coffea arabica (2n = 44). By comparing the RFLP patterns of potential diploid progenitor species with those of C. arabica, the sources of the two sets of chromosomes, or genomes, combined in C. arabica were identified. The genome organisation of C. arabica was confirmed by GISH using simultaneously labelled total genomic DNA from the two putative genome donor species as probes. These results clearly suggest that C. arabica is an amphidiploid formed by hybridisation between C. eugenioides and C. canephora, or ecotypes related to these diploid species. Our results also indicate low divergence between the two constituent genomes of C. arabica and those of its progenitor species, suggesting that the speciation of C. arabica took place relatively recently. Precise localisation in Central Africa of the site of the speciation of C. arabica, based on the present distribution of the coffee species, appears difficult, since the constitution and extent of tropical forest has varied considerably during the late Quaternary period.


Bacterial diversity in native habitats of the medicinal fungus Ophiocordyceps sinensis on Tibetan Plateau as determined using Illumina sequencing data.
Ophiocordyceps sinensis is one of the most well-known traditional Chinese medicinal fungi. In this study, bacterial diversity in the soils of native habitats of O. sinensis was investigated using Illumina sequencing data. A total of 525,000 sequences of V6-16S rRNA were analyzed. The number of OTUs from each sample ranged from 13,858 to 15,978 at 97% sequence similarity cut-off. The results demonstrated that the deep sequencing approach provides improved access to rare genotypes. Richness indices and Shannon's diversity index did not differ significantly between samples collected from locations where O. sinensis was present (Os1-3) and not present (NOs1-3). Classified bacterial sequences were grouped into 23 phyla including Proteobacteria, Actinobacteria, Acidobacteria, Verrucomicrobia, etc. The Venn diagram revealed that 7183 OTUs belonging to 14 phyla were shared by Os, NOs and MP (mycelial pellicle wrapping the sclerotium of O. sinensis) samples, possibly representing a core microbiome existing in native habitats of O. sinensis, and that 863 belonging to 12 phyla were shared by Os and MP samples, possibly related to the occurrence of O. sinensis. Overall, the results revealed a high bacterial diversity in the soil samples and the relationships between the bacterial diversity and O. sinensis merit further investigation.


An immunohistochemical study of the pathology of fatal malaria. Evidence for widespread endothelial activation and a potential role for intercellular adhesion molecule-1 in cerebral sequestration.
The sequestration of parasitized erythrocytes in the microvasculature of vital organs is central to the pathogenesis of severe Plasmodium falciparum malaria. This process is mediated by specific interactions between parasite adherence ligands and host receptors on vascular endothelium such as intercellular adhesion molecule-1 (ICAM-1) and CD36. Using immunohistochemistry we have examined the distribution of putative sequestration receptors in different organs from fatal cases of P. falciparum malaria and noninfected controls. Receptor expression and parasite sequestration in the brain were quantified and correlated. Fatal malaria was associated with widespread induction of endothelial activation markers, with significantly higher levels of ICAM-1 and E-selectin expression on vessels in the brain. In contrast, cerebral endothelial CD36 and thrombospondin staining were sparse, with no evidence for increased expression in malaria. There was highly significant co-localization of sequestration with the expression of ICAM-1, CD36, and E-selectin in cerebral vessels but no cellular inflammatory response. These results suggest that these receptors have a role in sequestration in vivo and indicate that systemic endothelial activation is a feature of fatal malaria.


Discovery of a magma chamber and faults beneath a Mid-Atlantic Ridge hydrothermal field.
Crust at slow-spreading ridges is formed by a combination of magmatic and tectonic processes, with magmatic accretion possibly involving short-lived crustal magma chambers. The reflections of seismic waves from crustal magma chambers have been observed beneath intermediate and fast-spreading centres, but it has been difficult to image such magma chambers beneath slow-spreading centres, owing to rough seafloor topography and associated seafloor scattering. In the absence of any images of magma chambers or of subsurface near-axis faults, it has been difficult to characterize the interplay of magmatic and tectonic processes in crustal accretion and hydrothermal circulation at slow-spreading ridges. Here we report the presence of a crustal magma chamber beneath the slow-spreading Lucky Strike segment of the Mid-Atlantic Ridge. The reflection from the top of the magma chamber, centred beneath the Lucky Strike volcano and hydrothermal field, is approximately 3 km beneath the sea floor, 3-4 km wide and extends up to 7 km along-axis. We suggest that this magma chamber provides the heat for the active hydrothermal vent field above it. We also observe axial valley bounding faults that seem to penetrate down to the magma chamber depth as well as a set of inward-dipping faults cutting through the volcanic edifice, suggesting continuous interactions between tectonic and magmatic processes.


Bone dimensional change with age: interactions of genetic, hormonal, and body size variables.
Changes in bony dimensions with age were assessed longitudinally from standardized X-ray films in 170 middle-aged Caucasian women, starting at age 40 years and with a median duration of observation of 21.125 years. Consistent with earlier work, cortical area of the metacarpals and radial shaft declined with age at rates ranging from 0.57 to 0.86%/year. As predicted, estrogen replacement therapy decreased this loss in a dose-dependent manner. Not previously reported is the fact that weight gain over the period of observation reduced upper extremity bone loss. Moreover, this protection was independent of the estrogen effect. In contrast with bone loss in the upper extremity, both femur shaft diameter and femur shaft cortical area increased significantly with age (0.23 and 0.26%/year, respectively). Estrogen replacement therapy inhibited femur shaft expansion but had no effect on femur cortical area. Weight change, however, strongly influenced gain (or loss) of femur cortical area: those in the highest weight change tertile gained 4 times as much cortical area as those in the lowest weight change tertile. VDR genotype also significantly influenced femoral shaft changes: women with the bb genotype had both greater shaft expansion and a greater increase in cortical area. The VDR effects were independent of the effects of weight change and estrogen. Femoral shaft expansion was of sufficient magnitude to suggest that the mechanical properties of the entire femur may change appreciably with age. Finally, contrary to widespread belief, there was significant, if modest, expansion at the femoral neck with age.


Dermal interstitial glucose as an indicator of ambient glycemia.
Using a novel minimally invasive (< or = 1.4 mm) technique to sample minuscule (0.5 microliter) amounts of dermal interstitial fluid (ISF), we assessed the accuracy of its glucose concentrations in predicting concurrently measured venous plasma and capillary plasma glucose concentrations. A total of 67 adult (37 male and 30 female) volunteers (57 with and 10 without diabetes) with venous plasma glucose levels from 1.6 to 28.4 mmol/l underwent forearm ISF, antecubetal venous, and fingertip capillary sampling. Rank correlations were 0.974 for ISF 1 vs. 2, 0.954 for ISF vs. venous, 0.935 for ISF vs. capillary, and 0.987 for venous vs. capillary. Median absolute differences were 0.53 mmol/l for ISF 1 vs. 2, 1.33 mmol/l for ISF vs. venous, 1.06 mmol/l for ISF vs. capillary, and 0.56 mmol/l for capillary vs. venous. Equations expressing ISF glucose as a function of venous and capillary glucose and equations expressing capillary glucose as a function of venous glucose had slopes of 0.995, 0.936, and 1.021, respectively (none significantly different from unity), and intercepts of 1.03 mmol/l (P = 0.024), 0.94 mmol/l (P = 0.131), and 0.56 mmol/l (P = 0.041), respectively. Error grid analysis of ISF vs. venous glucose and of capillary vs. venous glucose showed that 97% of the measurements fell within grids A and B. Dermal ISF sampling is a bloodless minimally invasive technique that provides a medium for glucose measurement, the concentrations of which closely reflect ambient glycemia to a degree comparable with that of capillary glucose measurements.


Hydrogel-incorporating unit in a well: 3D cell culture for high-throughput analysis.
The microfluidic 3D cell culture system has been an attractive model because it mimics the tissue and disease model, thereby expanding our ability to control the local cellular microenvironment. However, these systems still have limited value as quantitative assay tools due to the difficulties associated with the manipulation and maintenance of microfluidic cells, and their lack of compatibility with the high-throughput screening (HTS) analysis system. In this study, we suggest a microchannel-free, 3D cell culture system that has a hydrogel-incorporating unit integrated with a multi-well plate (24- to 96-well plate), which can provide better reproducibility in biological experiments. This plate was devised considering the design constraints imposed by various cell biology applications as well as by high-throughput analysis where the physical dimensions of the micro-features in the hydrogel-incorporating units were altered. We also demonstrated that the developed plate is potentially applicable to a variety of quantitative biochemical assays for qRT-PCR, Western blotting, and microplate-reader-based assays, such as ELISA, viability assay, and high content-screening (HCS) as well as the co-culture for biological studies. Human neural progenitor cells (hNPCs) that produce pathogenic Aβ species for modeling Alzheimer's disease (AD) were three-dimensionally cultured, and the efficacy of the inhibitors of Aβ production was assessed by ELISA in order to demonstrate the performance of this plate.


5-aminosalicylic acid improves lipid profile in mice fed a high-fat cholesterol diet through its dual effects on intestinal PPARγ and PPARα.
Obesity is associated with a series of metabolic complications, including dyslipidemia and insulin resistance (IR) that lack effective therapies. In recent years, intestinal inflammation has been suggested to contribute to obesity related metabolic syndrome and targeting gut inflammation with 5-ASA improves diet induced IR, however, its role in dyslipidemia is unknown and has never been explored. In the present study, we reported for the first time that administration of 5-ASA for 12 weeks significantly improved lipid profile by repressing plasma triglycerides and free cholesterol levels in mice fed high-fat cholesterol diet (HFC). In addition, liver lipids were significantly reduced by 5-ASA treatment in HFC-fed mice. Mechanistically, anti-inflammatory genes peroxisome proliferator-activated receptor-γ (Pparγ) and M2 marker, such as Mrc1 and Ym1, were remarkably upregulated, while pro-inflammation gene monocyte chemoattractant protein-1 (Mcp-1) were downregulated in small intestine of mice treated by 5-ASA. Further, 5-ASA improved gastrointestinal barrier by increasing the expression of the tight junction marker ZO-1. 5-ASA also enhanced cholesterol translocation by elevating genes expression of Npc1l1 and Abcg5/8. Moreover, mice fed HFC 5-ASA expressed increased Pparα in small intestinal and its target genes function in lipid oxidation and hydrolysis were remarkable elevated. Taken together, we reported a novel role of 5-ASA which may serve as a therapy target intestinal inflammation induced dyslipidemia.


A hierarchical glucose-intercalated NiMn-G-LDH@NiCo2S4 core-shell structure as a binder-free electrode for flexible all-solid-state asymmetric supercapacitors.
Flexible, lightweight, and high-energy-density asymmetric supercapacitors (ASCs) are highly attractive for portable and wearable electronics. However, the implementation of such flexible ASCs is still hampered by the low specific capacitance and sluggish reaction kinetics of the electrode materials. Herein, a hierarchical core-shell structure of hybrid glucose intercalated NiMn-LDH (NiMn-G-LDH)@NiCo2S4 hollow nanotubes is deliberately constructed on flexible carbon fiber cloth (CFC). The highly conductive hollow NiCo2S4 nanotube arrays can not only provide high-speed pathways for ion and electrolyte transfer but also regulate the growth of NiMn-G-LDH nanosheets. The expanded interlayer distance on NiMn-G-LDH nanosheets could further facilitate ion diffusion and improve the rate retention. Benefiting from the rational engineering, the flexible NiMn-G-LDH@NiCo2S4@CFC as a free-standing electrode could deliver a superior specific capacity of 1018 C g-1 at 1 A g-1, which is almost twice higher than that of pristine NiCo2S4@CFC. In addition, the as-assembled flexible all-solid-state ASC device (NiMn-G-LDH@NiCo2S4@CFC//AC) is capable of working at various bending angles and exhibits an impressive energy density of 60.3 W h kg-1 at a power density of 375 W kg-1, as well as a superior cycling stability of 86.4% after 10 000 cycles.


Unraveling the role of mitochondria during oxidative stress in plants.
The sedentary habit of plants means that they must stand and fight environmental stresses that their mobile animal cousins can avoid. A range of these abiotic stresses initiate the production in plant cells of reactive oxygen and nitrogen species that ultimately lead to oxidative damage affecting the yield and quality of plant products. A complex network of enzyme systems, producing and quenching these reactive species operate in different organelles. It is the integration of these compartmented defense systems that coordinates an effective response to the various stresses. Future attempts to improve plant growth or yield must consider the complexity of inter-organelle signaling and protein targeting if they are to be successful in producing plants with resistance to a broad range of stresses. Here we highlight the role of pre-oxidant, antioxidant, and post-oxidant defense systems in plant mitochondria and the potential role of proteins targeted to both mitochondria and chloroplasts, in an integrated defense against oxidative damage in plants.


Precision Medicine in Pancreatic Disease-Knowledge Gaps and Research Opportunities: Summary of a National Institute of Diabetes and Digestive and Kidney Diseases Workshop.
A workshop on research gaps and opportunities for Precision Medicine in Pancreatic Disease was sponsored by the National Institute of Diabetes and Digestive Kidney Diseases on July 24, 2019, in Pittsburgh. The workshop included an overview lecture on precision medicine in cancer and 4 sessions: (1) general considerations for the application of bioinformatics and artificial intelligence; (2) omics, the combination of risk factors and biomarkers; (3) precision imaging; and (4) gaps, barriers, and needs to move from precision to personalized medicine for pancreatic disease. Current precision medicine approaches and tools were reviewed, and participants identified knowledge gaps and research needs that hinder bringing precision medicine to pancreatic diseases. Most critical were (a) multicenter efforts to collect large-scale patient data sets from multiple data streams in the context of environmental and social factors; (b) new information systems that can collect, annotate, and quantify data to inform disease mechanisms; (c) novel prospective clinical trial designs to test and improve therapies; and (d) a framework for measuring and assessing the value of proposed approaches to the health care system. With these advances, precision medicine can identify patients early in the course of their pancreatic disease and prevent progression to chronic or fatal illness.


Electronic gaming and psychosocial adjustment.
The rise of electronic games has driven both concerns and hopes regarding their potential to influence young people. Existing research identifies a series of isolated positive and negative effects, yet no research to date has examined the balance of these potential effects in a representative sample of children and adolescents. The objective of this study was to explore how time spent playing electronic games accounts for significant variation in positive and negative psychosocial adjustment using a representative cohort of children aged 10 to 15 years. A large sample of children and adolescents aged 10 to 15 years completed assessments of psychosocial adjustment and reported typical daily hours spent playing electronic games. Relations between different levels of engagement and indicators of positive and negative psychosocial adjustment were examined, controlling for participant age and gender and weighted for population representativeness. Low levels (<1 hour daily) as well as high levels (>3 hours daily) of game engagement was linked to key indicators of psychosocial adjustment. Low engagement was associated with higher life satisfaction and prosocial behavior and lower externalizing and internalizing problems, whereas the opposite was found for high levels of play. No effects were observed for moderate play levels when compared with non-players. The links between different levels of electronic game engagement and psychosocial adjustment were small (<1.6% of variance) yet statistically significant. Games consistently but not robustly associated with children's adjustment in both positive and negative ways, findings that inform policy-making as well as future avenues for research in the area.