More mentoring needed? A cross-sectional study of mentoring programs for medical students in Germany.
Despite increasing recognition that mentoring is essential early in medical careers, little is known about the prevalence of mentoring programs for medical students. We conducted this study to survey all medical schools in Germany regarding the prevalence of mentoring programs for medical students as well as the characteristics, goals and effectiveness of these programs. A definition of mentoring was established and program inclusion criteria were determined based on a review of the literature. The literature defined mentoring as a steady, long-lasting relationship designed to promote the mentee's overall development. We developed a questionnaire to assess key characteristics of mentoring programs: the advocated mentoring model, the number of participating mentees and mentors, funding and staff, and characteristics of mentees and mentors (e.g., level of training). In addition, the survey characterized the mentee-mentor relationship regarding the frequency of meetings, forms of communication, incentives for mentors, the mode of matching mentors and mentees, and results of program evaluations. Furthermore, participants were asked to characterize the aims of their programs. The questionnaire consisted of 34 questions total, in multiple-choice (17), numeric (7) and free-text (10) format. This questionnaire was sent to deans and medical education faculty in Germany between June and September 2009. For numeric answers, mean, median, and standard deviation were determined. For free-text items, responses were coded into categories using qualitative free text analysis. We received responses from all 36 medical schools in Germany. We found that 20 out of 36 medical schools in Germany offer 22 active mentoring programs with a median of 125 and a total of 5,843 medical students (6.9 - 7.4% of all German medical students) enrolled as mentees at the time of the survey. 14 out of 22 programs (63%) have been established within the last 2 years. Six programs (27%) offer mentoring in a one-on-one setting. 18 programs (82%) feature faculty physicians as mentors. Nine programs (41%) involve students as mentors in a peer-mentoring setting. The most commonly reported goals of the mentoring programs include: establishing the mentee's professional network (13 programs, 59%), enhancement of academic performance (11 programs, 50%) and counseling students in difficulties (10 programs, 45%). Despite a clear upsurge of mentoring programs for German medical students over recent years, the overall availability of mentoring is still limited. The mentoring models and goals of the existing programs vary considerably. Outcome data from controlled studies are needed to compare the efficiency and effectiveness of different forms of mentoring for medical students.


Heart rate variability modifications following exercise training in type 2 diabetic patients with definite cardiac autonomic neuropathy.
Cardiac autonomic neuropathy (CAN) as a result of diabetic autonomic neuropathy is positively related to a poor prognosis in diabetic patients. The measurement of heart rate variability (HRV) is a remarkable index of cardiac autonomic dysfunction. The aim of this study was to examine the effects of long-term exercise training on HRV in type 2 diabetic patients with definite CAN. Seventeen type 2 diabetic patients with definite CAN (group A: 56.2 years (SD 5.8)) and 15 without CAN (group B: 55.8 years (SD 5.6)) participated in the study. All patients followed an aerobic exercise training programme three times a week for 6 months; the intensity of the session was 70% to 85% of heart rate reserve. At the beginning and end of the study all subjects underwent graded maximal exercise testing with spiroergometry for the evaluation of their aerobic capacity (VO(2)peak). Moreover, time and frequency domain indices of HRV were obtained from 24 h ambulatory continuous ECG Holter recordings. At baseline, all measurements of HRV indices were significantly reduced in group A compared with group B (p<0.05). Moreover, group A reached a significantly lower VO(2)peak by 14.8% compared with group B (p<0.05). Following the exercise training programme, the SD of all normal-to-normal RR intervals in the entire recording (SDNN) was increased by 18.8% (p<0.05) and 13.8% (p<0.05), the square root of the average of sum of squares of difference between adjacent filtered RR intervals (rMSSd) was increased by 35% (p<0.05) and 15.2% (p<0.05), and the percentage of differences between adjacent filtered RR intervals which was greater than 50 ms for the entire analysis (pNN50) was increased by 400% (p<0.05) and 67.9% (p<0.05) in groups A and B, respectively. Regarding the frequency domain indices, only the high frequency power (HF) was found to be significantly increased in group A. At the end of the exercise training programme, SDNN, rMSSd and low frequency power (LF) were significantly lower (24.3% (p<0.05), 20.3% (p<0.05) and 40% (p<0.05), respectively) in group A compared with group B. Also, VO(2)peak increased by 17.8% (p<0.05) in group A and by 11% (p<0.05) in group B. Furthermore, the exercise training programme had significant effects on blood lipid and glucose levels and glycosylated haemoglobin (HbA(1c)) in both groups. The results indicate that 6-month aerobic exercise training improves the cardiac autonomic nervous system function in type 2 diabetic patients. However, more favourable effects are found in type 2 diabetic patients with definite CAN.


Measurement of Adenovirus-Based Vector Heterogeneity.
Adenovirus vectors have become an important class of vaccines with the recent approval of Ebola and COVID-19 products. In-process quality attribute data collected during Adenovirus vector manufacturing has focused on particle concentration and infectivity ratios (based on viral genome: cell-based infectivity), and data suggest only a fraction of viral particles present in the final vaccine product are efficacious. To better understand this product heterogeneity, lab-scale preparations of two Adenovirus viral vectors, (Chimpanzee adenovirus (ChAdOx1) and Human adenovirus Type 5 (Ad5), were studied using transmission electron microscopy (TEM). Different adenovirus morphologies were characterized, and the proportion of empty and full viral particles were quantified. These proportions showed a qualitative correlation with the sample's infectivity values. Liquid chromatography-mass spectrometry (LC-MS) peptide mapping was used to identify key adenovirus proteins involved in viral maturation. Using peptide abundance analysis, a ∼5-fold change in L1 52/55k abundance was observed between low-(empty) and high-density (full) fractions taken from CsCl ultracentrifugation preparations of ChAdOx1 virus. The L1 52/55k viral protein is associated with DNA packaging and is cleaved during viral maturation, so it may be a marker for infective particles. TEM and LC-MS peptide mapping are promising higher-resolution analytical characterization tools to help differentiate between relative proportions of empty, non-infectious, and infectious viral particles as part of Adenovirus vector in-process monitoring, and these results are an encouraging initial step to better differentiate between the different product-related impurities.


Distinctive architecture of the chloroplast genome in the chlorophycean green alga Stigeoclonium helveticum.
The chloroplast genome has experienced many architectural changes during the evolution of chlorophyte green algae, with the class Chlorophyceae displaying the lowest degree of ancestral traits. We have previously shown that the completely sequenced chloroplast DNAs (cpDNAs) of Chamydomonas reinhardtii (Chlamydomonadales) and Scenedesmus obliquus (Sphaeropleales) are highly scrambled in gene order relative to one another. Here, we report the complete cpDNA sequence of Stigeoclonium helveticum (Chaetophorales), a member of a third chlorophycean lineage. This genome, which encodes 97 genes and contains 21 introns (including four putatively trans-spliced group II introns inserted at novel sites), is remarkably rich in derived features and extremely rearranged relative to its chlorophycean counterparts. At 223,902 bp, Stigeoclonium cpDNA is the largest chloroplast genome sequenced thus far, and in contrast to those of Chlamydomonas and Scenedesmus, features no large inverted repeat. Interestingly, the pattern of gene distribution between the DNA strands and the bias in base composition along each strand suggest that the Stigeoclonium genome replicates bidirectionally from a single origin. Unlike most known trans-spliced group II introns, those of Stigeoclonium exhibit breaks in domains I and II. By placing our comparative genome analyses in a phylogenetic framework, we inferred an evolutionary scenario of the mutational events that led to changes in genome architecture in the Chlorophyceae.


Positron insight into evolution of pore volume and penetration of the polymer network by n-heptane molecules in mesoporous XAD4.
The adsorption and desorption of n-heptane on the mesoporous polymer resin Amberlite XAD4 were investigated in situ by positron annihilation lifetime spectroscopy (PALS). This technique allows the monitoring of porosity and subnanometer free volume changes as well as the amount of liquid adsorbate captured within an investigated sorbent, without causing any interference with the course of adsorption/desorption. In consequence, the conducted studies provide microscale insight into the sorption processes of n-heptane (which is a significant component of volatile organic compounds - VOCs) on the polymeric material. The total pore volume decreases parabolically with n-heptane pressure until it reaches zero just below the saturated vapor pressure. Simultaneously, the average pore size increases linearly until it has approximately doubled. However, much faster rates of change in both these parameters occur at relative pressures below 0.05. The PALS results can be properly explained only if the swelling of the polymer skeleton is taken into account during the alkane adsorption process. This is confirmed by long-term pumping, which was required to achieve stabilization of PAL spectra during the final phase of desorption. In addition, the evolution of subnanometer free volumes (located between polymer chains and formed in liquid n-heptane) support this interpretation of the results.


Acupuncture for osteoarthritis of the knee: a systematic review.
To evaluate trials of acupuncture for osteoarthritis (OA) of the knee, to assess the methodologic quality of the trials and determine whether low-quality trials are associated with positive outcomes, to document adverse effects, to identify patient or treatment characteristics associated with positive response, and to identify areas of future research. Eight databases and 62 conference abstract series were searched. Randomized or quasi-randomized trials of all languages were included and evaluated for methodologic quality using the Jadad scale. Outcomes were pain, function, global improvement, and imaging. Data could not be pooled; therefore, a best-evidence synthesis was performed to determine the strength of evidence by control group. The adequacy of the acupuncture procedure was assessed by 2 acupuncturists trained in treating OA and blinded to study results. Seven trials representing 393 patients with knee OA were identified. For pain and function, there was limited evidence that acupuncture is more effective than being on a waiting list for treatment or having treatment as usual. For pain, there was strong evidence that real acupuncture is more effective than sham acupuncture; however, for function, there was inconclusive evidence that real acupuncture is more effective than sham acupuncture. There was insufficient evidence to determine whether the efficacy of acupuncture is similar to that of other treatments. The existing evidence suggests that acupuncture may play a role in the treatment of knee OA. Future research should define an optimal acupuncture treatment, measure quality of life, and assess acupuncture combined with other modalities.


Hypoxia in the androgen-dependent Shionogi model for prostate cancer at three stages.
The objective of this study was to investigate a possible relationship between androgen status and hypoxia in the Shionogi murine prostate tumor model, which is widely used to study the effects of androgen withdrawal on hormone resistance and radiation response. Binding of the nitroimidazole hypoxia marker EF5 was assessed using the Cy3-tagged monoclonal antibody ELK3-51. Three hours after injection of EF5 (30 mg/kg), tumors from the following three stages were excised: androgen-dependent, regressed tumors 7 days after castration, and androgen-independent. Half of each tumor was disaggregated for analysis by flow cytometry and the remainder was flash frozen. Statistically significant differences (P < 0.01) were found between androgen-dependent, regressed and androgen-dependent tumors: approximately 30, approximately 2 and approximately 50% hypoxic cells, respectively. Frozen sections from androgen-dependent tumors exhibited highly variable EF5 binding; regressed tumors showed very little or no binding; each section from androgen-dependent tumors showed high levels and uniformly distributed binding of EF5. There was no correlation between the degree of hypoxia and tumor weight (P > 0.1). The results from this preliminary study indicate that hypoxia may play an important role with respect to the timing of irradiation in prostate cancer treatments and possibly may be a useful prognostic tool. In addition, hypoxia may also be relevant to progression in this disease after androgen ablation.


Initiation of lung immunity: the afferent limb and the role of dendritic cells.
The lung is the largest epithelial surface in the body and constitutes a major portal of entry for microorganisms. Dendritic cells (DCs) are bone marrow-derived cells that populate the respiratory tract from the nasal mucosa to the lung pleura. DCs are dexterous cells playing a role in both the innate and the adaptive immune responses. DCs can respond to dangers in the lung by immediately generating protective cytokines that stimulate natural killer (NK) cells to produce cytokines and kill targets. Direct (by pathogen recognition receptors) and indirect (by inflammatory cytokines) recognition of the infectious agent can also trigger signaling pathways in the DCs that activate an integrated developmental program called maturation, which transforms DCs into efficient antigen-presenting cells (APCs) for naive T cell stimulation. Furthermore, DCs play a role in generating effector B cells in the lymph nodes draining the lung. This review focuses on how DCs sense the health of the lung and translate messages regarding the lung microenvironment to cells of the innate and adaptive immune response to initiate effector functions to clear an invading microorganism [T helper 1 (Th1) or Th2 response] or generate tolerance [T regulatory (Tr), anergy, or deletion] to inhaled antigen when no danger is sensed.


Coenzyme Q(10), vitamin E, selenium, and methionine in the treatment of chronic recurrent viral mucocutaneous infections.
Host defense and latency determinants in viral recurrent dermatologic infections are not entirely understood, as conventional protocols are inadequate to achieve fast healing and relapse prevention. Endogenously produced oxygen/nitrogen reactive species (ROS/RNS) are essential for antiviral immune defense, while their excess may aggravate skin inflammation. Here, we sought a nutritional approach capable of controlling ROS/RNS balance to accelerate recovery and inhibit recurrences of two mucocutaneous chronic DNA-virus infections. Two controlled clinical trials evaluated the feasibility of ROS/RNS-modulating nutriceutical dosages of coenzyme Q(10), RRR-α-tocopherol, selenium aspartate, and L-methionine associated with established therapies. Clinical trial 1 evaluated 68 patients with relapsing human papillomavirus skin warts treated with cryotherapy followed by 180 d of nutriceutical/placebo administration. Clinical trial 2 compared the combination of acyclovir followed by 90 d of nutriceutical administration versus acyclovir alone in patients with recurrences of herpes simplex genitalis (n = 60) or herpes zoster (n = 29). Viral DNA levels were assessed by polymer chain reaction, biomarkers of antiviral defense (peroxynitrite and IFNα/γ) and antioxidant capacity (lipophilic antioxidants and glutathione) were assayed by biochemical/enzyme-linked immunosorbent assay techniques in blood fractions. In both trials, the nutriceutical induced significantly faster healing (P < 0.01-0.05) with reduced incidence of relapses (P < 0.05) as compared to control groups, which was confirmed by decreased viral load and increased antiviral cytokine and peroxynitrite plasma levels. Plasma antioxidant capacity was higher (P < 0.01) in the experimental versus control groups. Results document positive clinical outcomes of the selected nutriceutical associated with conventional protocols in the management of relapsing mucocutaneous human papillomavirus and herpes infections.


The CD4/CD8 ratio in BAL fluid is highly variable in sarcoidosis.
Patients with pulmonary sarcoidosis frequently have increased numbers of lymphocytes and a high ratio of CD4+ to CD8+ T-lymphocytes (CD4/CD8 ratio) in bronchoalveolar lavage (BAL) fluid. Some investigators have suggested that these parameters can be used to distinguish sarcoidosis from other types of interstitial lung disease with a high degree of reliability. However, we hypothesized that the BAL CD4/CD8 ratio measured during the initial diagnostic evaluation of patients with biopsy-proven sarcoidosis is highly variable. BAL lymphocytes were analysed via flow cytometry to determine the CD4/CD8 ratio in a population of 86 patients with histological and clinical evidence of sarcoidosis, who underwent BAL as part of their initial diagnostic evaluation. In these patients, the CD4/CD8 ratio ranged 0.5-37.3, with a median value of 3.35 (mean 6.49). The CD4/CD8 ratio was greater than 4 in only 36 (42%) subjects. Ten patients (12%) had a CD4/CD8 ratio less than 1. The distribution of CD4/CD8 ratios was similar in the presence or absence of BAL lymphocytosis. In conclusion, the CD4/CD8 ratio in bronchoalveolar lavage fluid is highly variable in biopsy-proven sarcoidosis. Bronchoalveolar lavage lymphocyte subset determination is a diagnostic test with low sensitivity for this disease.


Anastomosing hemangioma of the liver and gastrointestinal tract: an unusual variant histologically mimicking angiosarcoma.
Anastomosing hemangioma, a benign vascular neoplasm histologically simulating angiosarcoma, is newly recognized and has been described primarily in the genitourinary tract. We have encountered this lesion in the liver and gastrointestinal tract, where it can be diagnostically challenging, especially in core biopsy. Herein, we described 6 cases of anastomosing hemangioma of the liver and gastrointestinal tract. They occurred in 4 women and 2 men, ranging in age from 48 to 71 years. The tumors ranged from 0.2 to 6 cm (median, 3.1 cm) and were grossly well demarcated with a gray-brown spongy appearance. Microscopically, they had a loosely lobulated architecture. At higher magnification, lesions consisted of anastomosing sinusoidal capillary-sized vessels with scattered hobnail endothelial cells. Mild cytologic atypia occurred in all cases. Mitoses were absent. Vascular thrombi were seen in 4 cases (66.7%) without necrosis. One tumor (16.7%) featured prominent extramedullary hematopoiesis and 1 (16.7%) hyaline globules. Immunohistochemistry results were available for 3 cases, and the lesions stained with CD34 and/or CD31. Five cases had clinical follow-up information; there were no recurrences or metastases (range, 8 to 96 mo; mean, 41 mo), and 1 patient received no follow-up after a benign diagnosis on her colon polyp. In summary, anastomosing hemangioma of the liver and gastrointestinal tract is a rare distinctive vascular neoplasm displaying overlapping features with well-differentiated angiosarcoma. Despite small numbers and limited follow-up information in our series, evidence to date supports that the lesion is benign. Awareness of this entity is essential to avoid overdiagnosis and unnecessary aggressive treatment.


Biolistic transfection of cultured myotubes.
Transfection of cells in culture with cDNA constructs is a powerful tool in cell biology, but postmitotic cells, including myotubes, can be hard to transfect with classic methods. Biolistics provides an alternative. We have used this biolistic technique to introduce cDNAs into cultured rat, chick, and C2C12 myotubes. This protocol results in efficient (20 to 70%, depending on cell type) transfection of myotubes, high levels of cDNA expression in individual myotubes, and little cellular damage. Using this procedure, we have expressed different muscle-specific cDNAs as green fluorescent protein (GFP) fusions. This technique is rapid, reliable, uses minimal amounts of reagent per transfection, and yields high transfection rates in a previously hard-to-transfect cell type. Its efficiency and reliability are high, regardless of plasmid size or epitope tag. Muscle cell biologists may now perform experiments in mature myotubes rather than relying on transfection of myoblast cultures or heterologous expression systems.


Effect of gentamicin and levels of ambient sound on hearing screening outcomes in the neonatal intensive care unit: A pilot study.
Hearing loss rates in infants admitted to neonatal intensive care units (NICU) run at 2-15%, compared to 0.3% in full-term births. The etiology of this difference remains poorly understood. We examined whether the level of ambient sound and/or cumulative gentamicin (an aminoglycoside) exposure affect NICU hearing screening results, as either exposure can cause acquired, permanent hearing loss. We hypothesized that higher levels of ambient sound in the NICU, and/or gentamicin dosing, increase the risk of referral on the distortion product otoacoustic emission (DPOAE) assessments and/or automated auditory brainstem response (AABR) screens. This was a prospective pilot outcomes study of 82 infants (<37 weeks gestational age) admitted to the NICU at Oregon Health & Science University. An ER-200D sound pressure level dosimeter was used to collect daily sound exposure in the NICU for each neonate. Gentamicin dosing was also calculated for each infant, including the total daily dose based on body mass (mg/kg/day), as well as the total number of treatment days. DPOAE and AABR assessments were conducted prior to discharge to evaluate hearing status. Exclusion criteria included congenital infections associated with hearing loss, and congenital craniofacial or otologic abnormalities. The mean level of ambient sound was 62.9 dBA (range 51.8-70.6 dBA), greatly exceeding American Academy of Pediatrics (AAP) recommendation of <45.0 dBA. More than 80% of subjects received gentamicin treatment. The referral rate for (i) AABRs, (frequency range: ∼1000-4000 Hz), was 5%; (ii) DPOAEs with a broad F2 frequency range (2063-10031 Hz) was 39%; (iii) DPOAEs with a low-frequency F2 range (<4172 Hz) was 29%, and (iv) DPOAEs with a high-frequency F2 range (>4172 Hz) was 44%. DPOAE referrals were significantly greater for infants receiving >2 days of gentamicin dosing compared to fewer doses (p = 0.004). The effect of sound exposure and gentamicin treatment on hearing could not be determined due to the low number of NICU infants without gentamicin exposure (for control comparisons). All infants were exposed to higher levels of ambient sound that substantially exceed AAP guidelines. More referrals were generated by DPOAE assessments than with AABR screens, with significantly more DPOAE referrals with a high-frequency F2 range, consistent with sound- and/or gentamicin-induced cochlear dysfunction. Adding higher frequency DPOAE assessments to existing NICU hearing screening protocols could better identify infants at-risk for ototoxicity.


Learning to Use the Force: Fitting Repulsive Potentials in Density-Functional Tight-Binding with Gaussian Process Regression.
The Density-Functional Tight Binding (DFTB) method is a popular semiempirical approximation to Density Functional Theory (DFT). In many cases, DFTB can provide comparable accuracy to DFT at a fraction of the cost, enabling simulations on length and time scales that are unfeasible with first-principles DFT. At the same time (and in contrast to empirical interatomic potentials and force fields), DFTB still offers direct access to electronic properties such as the band structure. These advantages come at the cost of introducing empirical parameters to the method, leading to a reduced transferability compared to true first-principle approaches. Consequently, it would be very useful if the parameter sets could be routinely adjusted for a given project. While fairly robust and transferable parametrization workflows exist for the electronic structure part of DFTB, the so-called repulsive potential Vrep poses a major challenge. In this paper, we propose a machine-learning (ML) approach to fitting Vrep, using Gaussian Process Regression (GPR) to reconstruct Vrep with DFT-DFTB force residues as training data. The use of GPR circumvents the need for nonlinear or global parameter optimization, while at the same time offering arbitrary flexibility in terms of the functional form. We also show that the proposed method can be applied to multiple elements at once, by fitting repulsive potentials for organic molecules containing carbon, hydrogen, and oxygen. Overall, the new approach removes focus from the choice of functional form and parametrization procedure, in favor of a data-driven philosophy.


Atypical lipomatous tumor of the tongue: report of a case.
The term atypical lipomatous tumor (ALT) is synonymous with well-differentiated liposarcoma (WDL). This tumor occurs very rarely in the tongue. Thus, it is difficult to predict its prognosis. Although recurrence of ALT/WDL is thought to be unlikely after complete excision, long-term follow-up is necessary when considering the pathologic conditions of this tumor at other sites. Here, we report a case of an ALT of the tongue, with a review of the literature. A 68-year-old man was referred to our hospital because of a tumor on the left side of his tongue. Upon palpation, the tumor was 12mm in diameter, circumscribed, elastic and hard, well demarcated, movable, and painless. We diagnosed the lesion as a lipoma and extirpated the tumor under local anesthesia. Because the specimen was histopathologically diagnosed as an ALT, as a precaution, we excised an additional 5mm from the area surrounding the original tumor under general anesthesia. Three years after the operation, the tongue demonstrated good healing without paresthesia or dysfunction, and to date there has been no evidence of recurrence.


Intertester reliability of shoulder complaints diagnoses in primary health care.
Shoulder complaints are frequently encountered in general practice, but precise diagnosing is challenging. This study investigated agreement of shoulder complaints diagnoses between clinicians in a primary health care setting. Cross-sectional study. Four primary health care clinicians used patients' history and functional examination of the shoulder by selective tissue tension techniques (STTs), to diagnose shoulder complaints. 62 patients, aged 18-75 years. Reliability of diagnoses was assessed by observed intertester agreement and Cohen's kappa. A total of 372 diagnostic pairs were available for intertester comparisons. Six diagnoses were assigned by all clinicians; supraspinatus-, infraspinatus-, subscapularis-tendinopathies; chronic subacromial bursitis; glenohumeral capsulitis, and acromioclavicular joint lesion. The observed agreement on these diagnoses ranged from 0.84 for glenohumeral capsulitis to 0.97 for acromioclavicular joint lesion. Kappa scores were 0.46 (95% CI 0.33, 0.58) for chronic subacromial bursitis; 0.53 (95% CI 0.34, 0.68), 0.59 (95% CI 0.47, 0.70), and 0.68 (95% CI 0.53, 0.82) for infraspinatus -, supraspinatus -, and subscapularis-tendinopathy, respectively. For glenohumeral capsulitis and acromioclavicular lesion kappa scores were 0.66 (95% CI 0.57, 0.73) and 0.78 (95% CI 0.61, 0.90). Kappa scores were higher for individual diagnoses than for individual tests, except for limitation in passive abduction (0.70, 95% CI 0.62, 0.78) and passive lateral rotation (0.66, 95% CI 0.57, 0.73). Although experienced clinicians showed substantial intertester agreement, precise diagnoses of shoulder complaints in primary health care remain a challenge. The present results call for further research on refined diagnoses of shoulder complaints. Key points Based on medical history and a systematic functional examination by selective tissue tension techniques (STTs), we investigated the agreement of shoulder complaints diagnoses across four primary health care clinicians and 62 patients. • Agreements on diagnoses were generally better than the agreement on individual tests. • Good kappa scores were obtained for the diagnoses glenohumeral capsulitis, rotator cuff tendinopathy, and acromioclavicular lesion. • Further research is necessary to investigate the diagnostic validity of functional shoulder examination by the STTs method.


Ocean acidification reduces induction of coral settlement by crustose coralline algae.
Crustose coralline algae (CCA) are a critical component of coral reefs as they accrete carbonate for reef structure and act as settlement substrata for many invertebrates including corals. CCA host a diversity of microorganisms that can also play a role in coral settlement and metamorphosis processes. Although the sensitivity of CCA to ocean acidification (OA) is well established, the response of their associated microbial communities to reduced pH and increased CO2 was previously not known. Here we investigate the sensitivity of CCA-associated microbial biofilms to OA and determine whether or not OA adversely affects the ability of CCA to induce coral larval metamorphosis. We experimentally exposed the CCA Hydrolithon onkodes to four pH/pCO2 conditions consistent with current IPCC predictions for the next few centuries (pH: 8.1, 7.9, 7.7, 7.5, pCO2 : 464, 822, 1187, 1638 μatm). Settlement and metamorphosis of coral larvae was reduced on CCA pre-exposed to pH 7.7 (pCO2 = 1187 μatm) and below over a 6-week period. Additional experiments demonstrated that low pH treatments did not directly affect the ability of larvae to settle, but instead most likely altered the biochemistry of the CCA or its microbial associates. Detailed microbial community analysis of the CCA revealed diverse bacterial assemblages that altered significantly between pH 8.1 (pCO2 = 464 μatm) and pH 7.9 (pCO2 = 822 μatm) with this trend continuing at lower pH/higher pCO2 treatments. The shift in microbial community composition primarily comprised changes in the abundance of the dominant microbes between the different pH treatments and the appearance of new (but rare) microbes at pH 7.5. Microbial shifts and the concomitant reduced ability of CCA to induce coral settlement under OA conditions projected to occur by 2100 is a significant concern for the development, maintenance and recovery of reefs globally.


Successful parenteral penicillin therapy of established Lyme arthritis.
In a double-blind placebo-controlled trial carried out from 1980 to 1982, 20 patients with established Lyme arthritis were assigned treatment with 2.4 million U of intramuscular benzathine penicillin weekly for three weeks (total, 7.2 million U) and 20 patients received saline. Seven of the 20 penicillin-treated patients (35 per cent) had complete resolution of arthritis soon after the injections and have remained well during a mean follow-up period of 33 months. In contrast, all 20 patients given placebo continued to have attacks of arthritis (P less than 0.02). In 1983, of 20 patients treated with intravenous penicillin G, 20 million U a day for 10 days, 11 (55 per cent) had complete resolution of arthritis and have remained well since. As compared with nonresponders, penicillin-responsive patients in both studies were more likely to have previously received antibiotics for erythema chronicum migrans (P less than 0.02) and less likely to have been given intraarticular corticosteroids during or at the conclusion of parenteral therapy (P less than 0.1). The Lyme spirochete was not cultured from synovium or joint fluid. We conclude that established Lyme arthritis can often be treated successfully with parenteral penicillin. However, neither of the regimens that we tested is uniformly effective, and further experience will be needed to determine the optimal course of therapy.


A multicouple probe for temperature gradient measurements in biological materials.
An easy-to-make, sensitive, thin, flexible, multisensor probe for in vivo tissue temperature profile measurement is described. It is essentially a multijunction thermocouple (i.e., a multicouple) of type-T composition. Enamel-insulated copper wires (38 gauge) were soldered 5 mm apart to one common uninsulated constantan wire (36 gauge) and introduced into a polyethylene tube sealed at one end. The total outside diameter of the multicouple probe is less than 1 mm, and the maximum number of junctions using the specified wire sizes is approximately 16. This design permits the instantaneous measurement of a tissue temperature profile at 5-mm intervals over a distance of approximately 8 cm. An extensive calibration for the thermal conductivity effect (k effect) along the multicouple wires by means of a limb model is presented. The results show that the temperature readings of the individual junctions are significantly affected by the k effect when a thermal gradient exists along the multicouple, as is usually the case during tissue temperature measurements. However, calibration of the multicouple for the k effect yields a measurement accuracy of +/- 0.1 degree C under a wide range of gradients. This probe can be implanted in tissues to measure thermal gradients under different physiological conditions.


Gli1 is a target of Sonic hedgehog that induces ventral neural tube development.
The vertebrate zinc finger genes of the Gli family are homologs of the Drosophila gene cubitus interruptus. In frog embryos, Gli1 is expressed transiently in the prospective floor plate during gastrulation and in cells lateral to the midline during late gastrula and neurula stages. In contrast, Gli2 and Gli3 are absent from the neural plate midline with Gli2 expressed widely and Gli3 in a graded fashion with highest levels in lateral regions. In mouse embryos, the three Gli genes show a similar pattern of expression in the neural tube but are coexpressed throughout the early neural plate. Because Gli1 is the only Gli gene expressed in prospective floor plate cells of frog embryos, we have investigated a possible involvement of this gene in ventral neural tube development. Here we show that Shh signaling activates Gli1 transcription and that widespread expression of endogenous frog or human glioma Gli1, but not Gli3, in developing frog embryos results in the ectopic differentiation of floor plate cells and ventral neurons within the neural tube. Floor-plate-inducing ability is retained when cytoplasmic Gli1 proteins are forced into the nucleus or are fused to the VP16 transactivating domain. Thus, our results identify Gli1 as a midline target of Shh and suggest that it mediates the induction of floor plate cells and ventral neurons by Shh acting as a transcriptional regulator.


Effects of excitatory amino acid antagonists on the phasic depolarizing events that occur in lumbar motoneurons during REM periods of active sleep.
The membrane potential of lumbar motoneurons is dominated during the tonic periods of active sleep by glycine-mediated inhibitory postsynaptic potentials (IPSPs). During the phasic rapid eye movement (REM) periods of active sleep there are also IPSPs but, in addition, the membrane potential exhibits depolarizing shifts and action potentials that occur in conjunction with the phasic activation of the somatic musculature. The present study was designed to provide evidence that an excitatory amino acid (EAA) neurotransmitter is responsible for these patterns of motoneuronal activation. It was found that juxtacellular microiontophoretic applications of kynurenic acid a non-NMDA antagonist of EAA neurotransmission, blocked the depolarizing potentials of motoneurons that arise during the REM periods of active sleep. In contrast, the selective NMDA receptor antagonist APV did not block these depolarizations. The conclusion is drawn that the myoclonic twitches and jerks that characterize that REM periods of active sleep are dependent upon the excitation of motoneurons that occurs as a result of EAAs acting at non-NMDA receptors.


BAYESIAN JOINT MODELING OF CHEMICAL STRUCTURE AND DOSE RESPONSE CURVES.
Today there are approximately 85,000 chemicals regulated under the Toxic Substances Control Act, with around 2,000 new chemicals introduced each year. It is impossible to screen all of these chemicals for potential toxic effects, either via full organism in vivo studies or in vitro high-throughput screening (HTS) programs. Toxicologists face the challenge of choosing which chemicals to screen, and predicting the toxicity of as yet unscreened chemicals. Our goal is to describe how variation in chemical structure relates to variation in toxicological response to enable in silico toxicity characterization designed to meet both of these challenges. With our Bayesian partially Supervised Sparse and Smooth Factor Analysis (BS3FA) model, we learn a distance between chemicals targeted to toxicity, rather than one based on molecular structure alone. Our model also enables the prediction of chemical dose-response profiles based on chemical structure (i.e., without in vivo or in vitro testing) by taking advantage of a large database of chemicals that have already been tested for toxicity in HTS programs. We show superior simulation performance in distance learning and modest to large gains in predictive ability compared to existing methods. Results from the high-throughput screening data application elucidate the relationship between chemical structure and a toxicity-relevant high-throughput assay. An R package for BS3FA is available online at https://github.com/kelrenmor/bs3fa.


Diode-pumped continuous wave and passively Q-switched Tm, Mg: LiTaO₃ lasers.
We have demonstrated the continuous wave and passively Q-switched Tm, Mg: LiTaO3 lasers for the first time. In continuous wave (CW) regime, a maximum CW output power of 1.03 W at 1952 nm was obtained, giving a slope efficiency of 9.5% and a beam quality M2 = 2.2. In passive Q-switching regime, a single walled carbon nanotube (SWCNT) was employed as saturable absorber (SA). The Tm,Mg:LiTaO3 laser has yielded a pulse of 560 ns under repetition rate of 34.2 kHz at 1926 nm, corresponding to a single pulse energy of 10.1 μJ. The results indicate a promising potential of nonlinear crystals in the applications for laser host materials.


Examining characteristics and associated distress related to Internet harassment: findings from the Second Youth Internet Safety Survey.
We sought to identify the characteristics of youth who are targets of Internet harassment and characteristics related to reporting distress as a result of the incident. The Second Youth Internet Safety Survey is a national telephone survey of a random sample of 1500 Internet users between the ages of 10 and 17 years conducted between March and June 2005. Participants had used the Internet at least once a month for the previous 6 months. Nine percent of the youth who used the Internet were targets of online harassment in the previous year. Thirty-two percent of the targets reported chronic harassment (ie, harassment > or = 3 times in the previous year). In specific incidents, almost half (45%) knew the harasser in person before the incident. Half of the harassers (50%) were reportedly male, and half (51%) were adolescents. One in 4 targets reported an aggressive offline contact (eg, the harasser telephoned, came to the youth's home, or sent gifts); 2 in 3 disclosed the incident to another person. Among otherwise similar youth, the odds of being a target of Internet harassment were higher for those youth who harassed others online, reported borderline/clinically significant social problems, and were victimized in other contexts. Likewise, using the Internet for instant messaging, blogging, and chat room use each elevated the odds of being a target of Internet harassment versus those who did not engage in these online activities. All other demographic, Internet-use, and psychosocial characteristics were not related to reports of online harassment. Thirty-eight percent of the harassed youth reported distress as a result of the incident. Those who were targeted by adults, asked to send a picture of themselves, received an aggressive offline contact (eg, the harasser telephoned or came to the youth's home), and were preadolescents were each significantly more likely to report distress because of the experience. Conversely, the youth who visited chat rooms were significantly less likely to be distressed by the harassment. Internet harassment can be a serious event for some youth. Because there has been a significant increase in the prevalence of Internet harassment from 2000 to 2005, adolescent health professionals should continue to be vigilant about such experiences in the lives of young people with whom they interact. Social problems and online aggressive behavior are each associated with elevated odds of being the target of harassment. Thus, prevention efforts may be best aimed at improving the interpersonal skills of young people who choose to communicate with others using these online tools. Adolescent health professionals should be especially aware of events that include aggressive offline contacts by adult harassers or asking the child or adolescent to send a picture of themselves, because each of these scenarios increase the odds of reporting distress by more than threefold. Findings further support the call for the inclusion of Internet-harassment prevention in conventional antibullying programs empowering schools to address Internet bullying situations that occur between students. This will not solve all situations, however. We also must encourage Internet service providers to partner with consumers to be proactive in serious harassment episodes that violate criminal laws and service-provider codes of conduct.


Comparative genomic analysis of Geobacter sulfurreducens KN400, a strain with enhanced capacity for extracellular electron transfer and electricity production.
A new strain of Geobacter sulfurreducens, strain KN400, produces more electrical current in microbial fuel cells and reduces insoluble Fe(III) oxides much faster than the wildtype strain, PCA. The genome of KN400 was compared to wildtype with the goal of discovering how the network for extracellular electron transfer has changed and how these two strains evolved. Both genomes were re-annotated, resulting in 14 fewer genes (net) in the PCA genome; 28 fewer (net) in the KN400 genome; and ca. 400 gene start and stop sites moved. 96% of genes in KN400 had clear orthologs with conserved synteny in PCA. Most of the remaining genes were in regions of genomic mobility and were strain-specific or conserved in other Geobacteraceae, indicating that the changes occurred post-divergence. There were 27,270 single nucleotide polymorphisms (SNP) between the genomes. There was significant enrichment for SNP locations in non-coding or synonymous amino acid sites, indicating significant selective pressure since the divergence. 25% of orthologs had sequence differences, and this set was enriched in phosphorylation and ATP-dependent enzymes. Substantial sequence differences (at least 12 non-synonymous SNP/kb) were found in 3.6% of the orthologs, and this set was enriched in cytochromes and integral membrane proteins. Genes known to be involved in electron transport, those used in the metabolic cell model, and those that exhibit changes in expression during growth in microbial fuel cells were examined in detail. The improvement in external electron transfer in the KN400 strain does not appear to be due to novel gene acquisition, but rather to changes in the common metabolic network. The increase in electron transfer rate and yield in KN400 may be due to changes in carbon flux towards oxidation pathways and to changes in ATP metabolism, both of which indicate that the overall energy state of the cell may be different. The electrically conductive pili appear to be unchanged, but cytochrome folding, localization, and redox potentials may all be affected, which would alter the electrical connection between the cell and the substrate.


Generation of pseudo-Bessel beams at THz frequencies by use of binary axicons.
In order to miniaturize and integrate conveniently in THz quasi-optical systems, binary axicons, based on binary optical ideas, are introduced in our paper and designed for generating pseudo-Bessel beams at THz frequencies. The designed binary axicons are easier to fabricate than holographic axicons, more compact and thus less lossy in the material when compared with classical cone axicons. To calculate the electromagnetic fields diffracted by binary axicons precisely, a two-dimension finite-difference time-domain (2-D FDTD) method in conjunction with Stratton- Chu formulas are employed in this paper. Applying this method, the properties of pseudo-Bessel beams produced by binary axicons are analyzed, and a brief summary is given in the end.


Postpartum Involution and Cancer: An Opportunity for Targeted Breast Cancer Prevention and Treatments?
Childbirth at any age confers a transient increased risk for breast cancer in the first decade postpartum and this window of adverse effect extends over two decades in women with late-age first childbirth (>35 years of age). Crossover to the protective effect of pregnancy is dependent on age at first pregnancy, with young mothers receiving the most benefit. Furthermore, breast cancer diagnosis during the 5- to 10-year postpartum window associates with high risk for subsequent metastatic disease. Notably, lactation has been shown to be protective against breast cancer incidence overall, with varying degrees of protection by race, multiparity, and lifetime duration of lactation. An effect for lactation on breast cancer outcome after diagnosis has not been described. We discuss the most recent data and mechanistic insights underlying these epidemiologic findings. Postpartum involution of the breast has been identified as a key mediator of the increased risk for metastasis in women diagnosed within 5-10 years of a completed pregnancy. During breast involution, immune avoidance, increased lymphatic network, extracellular matrix remodeling, and increased seeding to the liver and lymph node work as interconnected pathways, leading to the adverse effect of a postpartum diagnosis. We al discuss a novel mechanism underlying the protective effect of breastfeeding. Collectively, these mechanistic insights offer potential therapeutic avenues for the prevention and/or improved treatment of postpartum breast cancer.


Local environmental quality positively predicts breastfeeding in the UK's Millennium Cohort Study.
Background and Objectives: Breastfeeding is an important form of parental investment with clear health benefits. Despite this, rates remain low in the UK; understanding variation can therefore help improve interventions. Life history theory suggests that environmental quality may pattern maternal investment, including breastfeeding. We analyse a nationally representative dataset to test two predictions: (i) higher local environmental quality predicts higher likelihood of breastfeeding initiation and longer duration; (ii) higher socioeconomic status (SES) provides a buffer against the adverse influences of low local environmental quality. Methodology: We ran factor analysis on a wide range of local-level environmental variables. Two summary measures of local environmental quality were generated by this analysis-one 'objective' (based on an independent assessor's neighbourhood scores) and one 'subjective' (based on respondent's scores). We used mixed-effects regression techniques to test our hypotheses. Results: Higher objective, but not subjective, local environmental quality predicts higher likelihood of starting and maintaining breastfeeding over and above individual SES and area-level measures of environmental quality. Higher individual SES is protective, with women from high-income households having relatively high breastfeeding initiation rates and those with high status jobs being more likely to maintain breastfeeding, even in poor environmental conditions. Conclusions and Implications: Environmental quality is often vaguely measured; here we present a thorough investigation of environmental quality at the local level, controlling for individual- and area-level measures. Our findings support a shift in focus away from individual factors and towards altering the landscape of women's decision making contexts when considering behaviours relevant to public health.


Capillary Rarefaction Associates with Albuminuria: The Maastricht Study.
Albuminuria may be a biomarker of generalized (i.e., microvascular and macrovascular) endothelial dysfunction. According to this concept, endothelial dysfunction of the renal microcirculation causes albuminuria by increasing glomerular capillary wall permeability and intraglomerular pressure, the latter eventually leading to glomerular capillary dropout (rarefaction) and further increases in intraglomerular pressure. However, direct evidence for an association between capillary rarefaction and albuminuria is lacking. Therefore, we examined the cross-sectional association between the recruitment of capillaries after arterial occlusion (capillary density during postocclusive peak reactive hyperemia) and during venous occlusion (venous congestion), as assessed with skin capillaroscopy, and albuminuria in 741 participants of the Maastricht Study, including 211 participants with type 2 diabetes. Overall, 57 participants had albuminuria, which was defined as a urinary albumin excretion ≥30 mg/24 h. After adjustment for potential confounders, participants in the lowest tertile of skin capillary recruitment during postocclusive peak reactive hyperemia had an odds ratio for albuminuria of 2.27 (95% confidence interval, 1.07 to 4.80) compared with those in the highest tertile. Similarly, a comparison between the lowest and the highest tertiles of capillary recruitment during venous congestion yielded an odds ratio of 2.89 (95% confidence interval, 1.27 to 6.61) for participants in the lowest tertile. In conclusion, lower capillary density of the skin microcirculation independently associated with albuminuria, providing direct support for a role of capillary rarefaction in the pathogenesis of albuminuria.


Sentinel events predicting later unwanted sex among girls: A national survey in Haiti, 2012.
Sexual violence against children is a significant global public health problem, yet limited studies exist from low-resource settings. In Haiti we conducted the country's first, nationally representative survey focused on childhood violence to help inform the development of a national action plan for violence against children. The Haiti Violence Against Children Survey was a household-level, multistage, cluster survey among youth age 13-24. In this analysis we sought to determine whether sexual violence sentinel events (unwanted sexual touching or unwanted attempted sex) were predictive of later unwanted, completed, penetrative sex in Haiti. We also sought to explore characteristics of sentinel events and help-seeking behavior among Haitian children. Multivariable logistic regression was used to test associations between sentinel events and later unwanted, completed, penetrative sex. Overall, 1,457 females reported on experiences of sexual violence occurring in childhood (before age 18). A sentinel event occurred in 40.4% of females who experienced subsequent unwanted completed sex. Females experiencing a sentinel event were approximately two and a half times more likely to experience later unwanted completed sex (adjusted odds ratio=2.40, p=.004) compared to individuals who did not experience a sentinel event. The mean lag time from first sentinel event to first unwanted completed sex was 2.3 years. Only half (54.6%) of children experiencing a sentinel event told someone about their experience of sexual violence. Among children, sentinel events occur frequently before later acts of completed unwanted sex and may represent a useful point of intervention. Reporting of sexual violence by children in Haiti is low and can be improved to better act on sentinel events.


Bat flight generates complex aerodynamic tracks.
The flapping flight of animals generates an aerodynamic footprint as a time-varying vortex wake in which the rate of momentum change represents the aerodynamic force. We showed that the wakes of a small bat species differ from those of birds in some important respects. In our bats, each wing generated its own vortex loop. Also, at moderate and high flight speeds, the circulation on the outer (hand) wing and the arm wing differed in sign during the upstroke, resulting in negative lift on the hand wing and positive lift on the arm wing. Our interpretations of the unsteady aerodynamic performance and function of membranous-winged, flapping flight should change modeling strategies for the study of equivalent natural and engineered flying devices.


A 12-week, open, randomized trial comparing sodium valproate to lithium in patients with bipolar I disorder suffering from a manic episode.
On the basis of 3-week studies, lithium and valproate are both recommended for first-line treatment of acute mania. It is, however, also important to demonstrate that antimanic efficacy can be maintained. This study has been designed to compare the efficacy and tolerability of valproate and lithium over 12 weeks in the treatment of acute mania in patients with type I bipolar disorder. Three hundred patients with bipolar I disorder presenting with acute mania were randomized to open treatment with lithium (starting dose: 400 mg/day) or valproate (starting dose: 20 mg/kg/day) for 12 weeks. The primary efficacy criterion was remission (YMRS score <or=12 at study end and a reduction of >or=2 on the CGI-BP severity scale). Remission rates were 65.5% (lithium group) and 72.3% (valproate group). Noninferiority of valproate with respect to lithium was demonstrated [between-group difference: 6.78% (95% confidence intervals: -3.80 to 17.36%)]. Remission rates assessed by the secondary mixed model repeated measures analysis were significantly greater with valproate than with lithium. Adverse events were reported in 44% of patients in both groups. Valproate and lithium showed comparable efficacy and tolerability in the treatment of acute mania over 12 weeks.


An Adaptive Prediction-Based Approach to Lossless Compression of Floating-Point Volume Data.
In this work, we address the problem of lossless compression of scientific and medical floating-point volume data. We propose two prediction-based compression methods that share a common framework, which consists of a switched prediction scheme wherein the best predictor out of a preset group of linear predictors is selected. Such a scheme is able to adapt to different datasets as well as to varying statistics within the data. The first method, called APE (Adaptive Polynomial Encoder), uses a family of structured interpolating polynomials for prediction, while the second method, which we refer to as ACE (Adaptive Combined Encoder), combines predictors from previous work with the polynomial predictors to yield a more flexible, powerful encoder that is able to effectively decorrelate a wide range of data. In addition, in order to facilitate efficient visualization of compressed data, our scheme provides an option to partition floating-point values in such a way as to provide a progressive representation. We compare our two compressors to existing state-of-the-art lossless floating-point compressors for scientific data, with our data suite including both computer simulations and observational measurements. The results demonstrate that our polynomial predictor, APE, is comparable to previous approaches in terms of speed but achieves better compression rates on average. ACE, our combined predictor, while somewhat slower, is able to achieve the best compression rate on all datasets, with significantly better rates on most of the datasets.


Sugar nucleotide quantification by liquid chromatography tandem mass spectrometry reveals a distinct profile in Plasmodium falciparum sexual stage parasites.
The obligate intracellular lifestyle of Plasmodium falciparum and the difficulties in obtaining sufficient amounts of biological material have hampered the study of specific metabolic pathways in the malaria parasite. Thus, for example, the pools of sugar nucleotides required to fuel glycosylation reactions have never been studied in-depth in well-synchronized asexual parasites or in other stages of its life cycle. These metabolites are of critical importance, especially considering the renewed interest in the presence of N-, O-, and other glycans in key parasite proteins. In this work, we adapted a liquid chromatography tandem mass spectrometry (LC-MS/MS) method based on the use of porous graphitic carbon (PGC) columns and MS-friendly solvents to quantify sugar nucleotides in the malaria parasite. We report the thorough quantification of the pools of these metabolites throughout the intraerythrocytic cycle of P. falciparum The sensitivity of the method enabled, for the first time, the targeted analysis of these glycosylation precursors in gametocytes, the parasite sexual stages that are transmissible to the mosquito vector.


Phospholipase A2IVα regulates phagocytosis independent of its enzymatic activity.
Group IVα phospholipase A(2) (PLA(2)IVα) is a lipolytic enzyme that catalyzes the hydrolysis of membrane phospholipids to generate precursors of potent inflammatory lipid mediators. Here, the role of PLA(2)IVα in Fc receptor (FcR)-mediated phagocytosis was investigated, demonstrating that PLA(2)IVα is selectively activated upon FcR-mediated phagocytosis in macrophages and that it rapidly translocates to the site of the nascent phagosome. Moreover, pharmacological inhibition of PLA(2)IVα by pyrrophenone reduces particle internalization by up to 50%. In parallel, fibroblasts from PLA(2)IVα knock-out mice overexpressing FcγRIIA and able to internalize IgG-opsonized beads show 50% lower phagocytosis, compared with wild-type cells, and transfection of PLA(2)IVα fully recovers this impaired function. Interestingly, transfection of the catalytically inactive deleted PLA(2)IVα mutant (PLA(2)IVα(1-525)) and point mutant (PLA(2)IVα-S228C) also promotes recovery of this impaired function. Finally, transfection of the PLA(2)IVα C2 domain (which is directly involved in PLA(2)IVα membrane binding), but not of PLA(2)IVα-D43N (which cannot bind to membranes), rescues FcR-mediated phagocytosis. These data unveil a new mechanism of action for PLA(2)IVα, which demonstrates that the membrane binding, and not the enzymatic activity, is required for PLA(2)IVα modulation of FcR-mediated phagocytosis.


Deadenylase depletion protects inherited mRNAs in primordial germ cells.
A crucial event in animal development is the specification of primordial germ cells (PGCs), which become the stem cells that create sperm and eggs. How PGCs are created provides a valuable paradigm for understanding stem cells in general. We find that the PGCs of the sea urchin Strongylocentrotus purpuratus exhibit broad transcriptional repression, yet enrichment for a set of inherited mRNAs. Enrichment of several germline determinants in the PGCs requires the RNA-binding protein Nanos to target the transcript that encodes CNOT6, a deadenylase, for degradation in the PGCs, thereby creating a stable environment for RNA. Misexpression of CNOT6 in the PGCs results in their failure to retain Seawi transcripts and Vasa protein. Conversely, broad knockdown of CNOT6 expands the domain of Seawi RNA as well as exogenous reporters. Thus, Nanos-dependent spatially restricted CNOT6 differential expression is used to selectively localize germline RNAs to the PGCs. Our findings support a 'time capsule' model of germline determination, whereby the PGCs are insulated from differentiation by retaining the molecular characteristics of the totipotent egg and early embryo.


Assessing outcomes in palliative chemotherapy.
Patients with metastatic cancer are often offered systemic chemotherapy, although it is not usually curative in this situation. The impact of treatment on survival may be small and the intention is mainly to palliate symptoms and to improve the patient's quality of life. Traditional end points in clinical trials tend to focus on cancer outcomes, such as tumour shrinkage or survival. Trials of palliative chemotherapy should focus on patient outcomes that reflect the aims of treatment. There are now a large number of instruments available to measure quality of life and the difficulty lies in deciding which tools are appropriate and applying them consistently to give results that are robust enough to enable clinical decision making. This is particularly challenging in a population of patients with a high degree of morbidity. In addition, economic end points are becoming increasingly relevant in the current climate: as newer treatments become available, the ways in which costs may be balanced against the potential benefits in terms of gains in quality of life become important.


Schizophrenia and the frontal lobes: post-mortem stereological study of tissue volume.
It has been suggested that there is frontal lobe involvement in schizophrenia, and that it may be lateralised and gender-specific. To clarify the structure of the frontal lobes in schizophrenia in a post-mortem series. The volume of white matter and cortical components of the frontal lobes was measured in brains of controls and patients with schizophrenia using planimetry and the Cavalieri principle. The components measured were: superior frontal gyrus, middle frontal gyrus, a composite of inferior frontal gyrus and orbito-frontal cortex, as well as total frontal lobe cortex and white matter. In addition, the anterior cingulate gyrus was measured. No diagnosis, gender, diagnosis x side, diagnosis x gender or diagnosis x gender x side interactions were observed in the volume of any of the components, the grey matter as a whole or the white matter. No evidence for volumetric inter-group differences was found for the anterior cingulate gyrus. Such structural abnormalities as are present in the frontal lobes are more subtle than straightforward alterations in tissue volume; they may include changes in shape and the pattern of gyral folding.


Building black holes: supercomputer cinema.
A new computer code can solve Einstein's equations of general relativity for the dynamical evolution of a relativistic star cluster. The cluster may contain a large number of stars that move in a strong gravitational field at speeds approaching the speed of light. Unstable star clusters undergo catastrophic collapse to black holes. The collapse of an unstable cluster to a supermassive black hole at the center of a galaxy may explain the origin of quasars and active galactic nuclei. By means of a supercomputer simulation and color graphics, the whole process can be viewed in real time on a movie screen.


Relationships between B cell and myeloid differentiation. Studies with a B lymphocyte progenitor line, HAFTL-1.
A cell line, HAFTL-1, derived by in vitro transformation of fetal liver cells with v-Ha-ras, was found to have molecular and phenotypic characteristics of pro-B cells recently committed to the Ly-1+ B cell differentiation pathway. Stimulation of these cells with LPS resulted in their differentiation within either the B or myelomonocytic lineages. Thus, lines derived from LPS-stimulated HAFTL-1 cells were shown to be clonally related, as evidenced by common v-ras integrations, but to exhibit characteristics of pre-B cells (ThB expression, continuing DJ heavy chain rearrangements) or mature macrophages (expression of Mac-1 and Mac-2, lysozyme and nonspecific esterase production, phagocytosis) while maintaining their Ly-1+ phenotype. These results suggest that events resulting in the irrevocable commitment to a single lineage occur late in differentiation, at least within the pathway yielding Ly-1+ B cells and a proposed subpopulation of Ly-1+ monocytes and macrophages. Final commitment to these lineages is carefully orchestrated, as evidenced by restricted expression of Ly-5 isoforms and production of IgH transcripts.


Methods for identifying and assessing groups in health behavioral research.
Methods to study groups developed in other fields have much to contribute to the study of health behavior, including alcohol and substance use. This paper surveys group methods in order to demonstrate how attention to group levels of analysis can provide new insights about the context and causes of health behavior. The first major section of this paper focuses on the influences on health behavior associated with "naturally occurring groups" such as peer networks, families, workplaces, neighborhoods, churches, treatment groups and residential settings. Different aspects of group influence are discussed in terms of seven different features of social groups: history, structure, function, resources, process, norms and climate. The second major part of this paper is concerned with methods to identify meaningful subgroups in heterogeneous samples. Following an overview of taxonomic methods to identify and assign cases to groups, we highlight the potential for these methods to help address a number of fundamental theoretical problems in health behavioral research, including: describing cultural diversity, distinguishing response sets in survey research, controlling error in outcomes scores related to baseline differences, tracking trajectories of change, and revealing interactions among determinants of health behavior. The third and final section of the paper highlights specific applications of taxonomic and group-influence methodologies in intervention research, including design and analysis of randomized trials, studies of existing treatment settings and group-level variables involved in translational research.


Management of Botrytis Leaf Blight of Onion: The Québec Experience of 20 Years of Continual Improvement.
Botrytis leaf blight (BLB) of onion (Allium cepa) is caused by Botrytis squamosa. The disease has been reported on onion crops in several of the onion production areas of the world including North and South America, Europe, Asia, and Australia, although it is not a problem in arid production regions such as the western United States. In eastern Canada, the disease is generally present every year and is especially severe on cultivars of yellow globe onion. The pathogen biology and disease epidemiology have been intensively researched. Over the last few decades, in the organic soil area of Quebec, extensive research effort has been devoted to the development and evaluation of predictive models and disease management strategies. There has been an active integrated pest management program for onions since the early 1980s, and scouting for disease has played a major role in disease management. In this article, the story of BLB management in eastern Canada over a period of two decades is summarized.


A basic helix-loop-helix transcription factor controls cell growth and size in root hairs.
Postmitotic cell growth defines cell shape and size during development. However, the mechanisms regulating postmitotic cell growth in plants remain unknown. Here we report the discovery of a basic helix-loop-helix (bHLH) transcription factor called RSL4 (ROOT HAIR DEFECTIVE 6-LIKE 4) that is sufficient to promote postmitotic cell growth in Arabidopsis thaliana root-hair cells. Loss of RSL4 function resulted in the development of very short root hairs. In contrast, constitutive RSL4 expression programmed constitutive growth, resulting in the formation of very long root hairs. Hair-cell growth signals, such as auxin and low phosphate availability, modulate hair cell extension by regulating RSL4 transcript and protein levels. RSL4 is thus a regulator of growth that integrates endogenous developmental and exogenous environmental signals that together control postmitotic growth in root hairs. The control of postmitotic growth by transcription factors may represent a general mechanism for regulating cell size across diverse organisms.


Tocotrienols, the vitamin E of the 21st century: its potential against cancer and other chronic diseases.
Initially discovered in 1938 as a "fertility factor," vitamin E now refers to eight different isoforms that belong to two categories, four saturated analogues (α, β, γ, and δ) called tocopherols and four unsaturated analogues referred to as tocotrienols. While the tocopherols have been investigated extensively, little is known about the tocotrienols. Very limited studies suggest that both the molecular and therapeutic targets of the tocotrienols are distinct from those of the tocopherols. For instance, suppression of inflammatory transcription factor NF-κB, which is closely linked to tumorigenesis and inhibition of HMG-CoA reductase, mammalian DNA polymerases and certain protein tyrosine kinases, is unique to the tocotrienols. This review examines in detail the molecular targets of the tocotrienols and their roles in cancer, bone resorption, diabetes, and cardiovascular and neurological diseases at both preclinical and clinical levels. As disappointment with the therapeutic value of the tocopherols grows, the potential of these novel vitamin E analogues awaits further investigation.


The reasons for discontinuation of infliximab treatment in patients with Crohn's disease: a review of practice at NHS teaching hospital.
Introduction. There is little information on the reasons for discontinuing infliximab treatment in patients with Crohn's disease. The aim of this study was to document these reasons to determine if any were preventable which would allow patients to continue the therapy. Aims & Methods. A review of the medical notes was conducted at the Norfolk and Norwich University Hospital on patients with Crohn's disease treated with infliximab between 2002-2008 to determine the reasons for stopping it. Results. A total of 65 patients were identified who had treatment with infliximab, of whom 23 (35.3%) had their therapy stopped. The reasons for discontinuation of infliximab in the 23 patients were: 47.8% side effects, 17.4% refractory disease, 13.0% achieved remission and did not receive long-term maintenance treatment, 4.34% pregnancy, 4.34% death, and unknown 13.0%. Conclusions. The main reasons for the discontinuation of infliximab were side effects rather than a lack of clinical response.


Risk of asthma and autoimmune diseases and related conditions in patients hospitalized for obesity.
Although there are putative mechanistic links between obesity and autoimmune diseases, obesity is not considered a risk factor for most autoimmune diseases. Using the nation-wide Hospital Discharge Register we defined a cohort of 29,665 patients hospitalized for obesity since year 1964. The patients were followed for hospitalization for any of 34 autoimmune or related conditions through year 2007. Standardized incidence ratios (SIRs) were calculated for autoimmune diseases in obese individuals compared to those who had not been hospitalized for obesity. Among 22 immune diseases diagnosed after hospitalization for obesity and in at least 5 patients, the overall SIR was 2.05. Of the individual diseases studied, the risk of 16 was significantly increased; none displayed a decreased risk. Psoriasis (4.54) and Behçet's disease (4.49) exhibited the highest risks, followed by Hashimoto's disease/hypothyroidism (4.12) and asthma (3.39). Small but significant increases in SIRs were also noted for the common autoimmune diseases Graves's disease/hyperthyroidism (1.28) and rheumatoid arthritis (1.37). The present population of obese individuals, subsequently diagnosed with a number of autoimmune diseases and related conditions, was hospitalized at a relatively young age. Further studies are needed to describe the morbidity in the obese population at large.


KIFC1 is a novel potential therapeutic target for breast cancer.
Kinesin-like protein KIFC1, a normally nonessential kinesin motor, plays a critical role in centrosome clustering in cancer cells and is essential for the survival of cancer cells. Herein, we reported that KIFC1 expression is up-regulated in breast cancer, particularly in estrogen receptor negative, progesterone receptor negative and triple negative breast cancer, and is not associated with epidermal growth factor receptor 2 status. In addition, KIFC1 is highly expressed in all 8 tested human breast cancer cell lines, but is absent in normal human mammary epithelial cells and weakly expressed in 2 human lung fibroblast lines. Moreover, KIFC1 silencing significantly reduced breast cancer cell viability. Finally, we found that PJ34, a potent small molecule inhibitor of poly(ADP-ribose) polymerase, suppressed KIFC1 expression and induced multipolar spindle formation in breast cancer cells, and inhibited cell viability and colony formation within the same concentration range, suggesting that KIFC1 suppression by PJ34 contributes to its anti-breast cancer activity. Together, these results suggest that KIFC1 is a novel promising therapeutic target for breast cancer.


The harmonizome: a collection of processed datasets gathered to serve and mine knowledge about genes and proteins.
Genomics, epigenomics, transcriptomics, proteomics and metabolomics efforts rapidly generate a plethora of data on the activity and levels of biomolecules within mammalian cells. At the same time, curation projects that organize knowledge from the biomedical literature into online databases are expanding. Hence, there is a wealth of information about genes, proteins and their associations, with an urgent need for data integration to achieve better knowledge extraction and data reuse. For this purpose, we developed the Harmonizome: a collection of processed datasets gathered to serve and mine knowledge about genes and proteins from over 70 major online resources. We extracted, abstracted and organized data into ∼72 million functional associations between genes/proteins and their attributes. Such attributes could be physical relationships with other biomolecules, expression in cell lines and tissues, genetic associations with knockout mouse or human phenotypes, or changes in expression after drug treatment. We stored these associations in a relational database along with rich metadata for the genes/proteins, their attributes and the original resources. The freely available Harmonizome web portal provides a graphical user interface, a web service and a mobile app for querying, browsing and downloading all of the collected data. To demonstrate the utility of the Harmonizome, we computed and visualized gene-gene and attribute-attribute similarity networks, and through unsupervised clustering, identified many unexpected relationships by combining pairs of datasets such as the association between kinase perturbations and disease signatures. We also applied supervised machine learning methods to predict novel substrates for kinases, endogenous ligands for G-protein coupled receptors, mouse phenotypes for knockout genes, and classified unannotated transmembrane proteins for likelihood of being ion channels. The Harmonizome is a comprehensive resource of knowledge about genes and proteins, and as such, it enables researchers to discover novel relationships between biological entities, as well as form novel data-driven hypotheses for experimental validation.Database URL: http://amp.pharm.mssm.edu/Harmonizome.


High temperature X-ray diffraction and thermo-gravimetrical analysis of the cubic perovskite Ba0.5Sr0.5Co0.8Fe0.2O3-δ under different atmospheres.
Ba(0.5)Sr(0.5)Co(0.8)Fe(0.2)O(3-δ) (BSCF) with the cubic perovskite structure is known to be metastable at low temperature under an oxidizing atmosphere. Here, the thermal and chemical expansion of BSCF were studied by in situ high temperature powder X-ray diffraction and thermo-gravimetrical analysis (TGA) in partial pressure of oxygen ranging from an inert atmosphere (∼10(-4) bar) to 10 bar O(2). The BSCF powder, heat treated at 1000 °C and quenched to ambient temperature prior to the analysis, was shown to oxidize under an oxidizing atmosphere before thermal reduction took place. With decreasing partial pressure of oxygen the initial oxidation was suppressed and only reduction of Co/Fe and loss of oxygen were observed under an inert atmosphere. The thermal expansion of BSCF under different atmospheres was determined from the thermal evolution of the cubic unit cell parameter, demonstrating that the thermal expansion of BSCF depends on the atmosphere. Chemical expansion of BSCF was also estimated based on the diffraction data and thermo-gravimetrical analysis. A hexagonal perovskite phase, coexisting with the cubic BSCF polymorph, was observed to be formed above 600 °C during heating. The phase separation leading to the formation of the hexagonal polymorph was driven by oxidation, and the unit cell of the cubic BSCF was shown to decrease with increasing amounts of the hexagonal phase. The hexagonal phase disappeared upon further heating, accompanied with an expansion of the unit cell of the cubic BSCF.


Distinct patterns of neural activity during memory formation of nonwords versus words.
Research into the neural underpinnings of memory formation has focused on the encoding of familiar verbal information. Here, we address how the brain supports the encoding of novel information that does not have meaning. Electrical brain activity was recorded from the scalps of healthy young adults while they performed an incidental encoding task (syllable judgments) on separate series of words and "nonwords" (nonsense letter strings that are orthographically legal and pronounceable). Memory for the items was then probed with a recognition memory test. For words as well as nonwords, event-related potentials differed depending on whether an item would subsequently be remembered or forgotten. However, the polarity and timing of the effect varied across item type. For words, subsequently remembered items showed the usually observed positive-going, frontally distributed modulation from around 600 msec after word onset. For nonwords, by contrast, a negative-going, spatially widespread modulation predicted encoding success from 1000 msec onward. Nonwords also showed a modulation shortly after item onset. These findings imply that the brain supports the encoding of familiar and unfamiliar letter strings in qualitatively different ways, including the engagement of distinct neural activity at different points in time. The processing of semantic attributes plays an important role in the encoding of words and the associated positive frontal modulation.


Aqueous humor metabolite profile of pseudoexfoliation glaucoma is distinctive.
Pseudoexfoliation (PEX) is a known cause of secondary open angle glaucoma. PEX glaucoma is associated with structural and metabolic changes in the eye. Despite similarities, PEX and primary open angle glaucoma (POAG) may have differences in the composition of metabolites. We analyzed the metabolites of the aqueous humor (AH) of PEX subjects sequentially first using nuclear magnetic resonance (1H NMR: HSQC and TOCSY), and subsequently with liquid chromatography tandem mass spectrometry (LC-MS/MS) implementing isotopic ratio outlier analysis (IROA) quantification. The findings were compared with previous results for POAG and control subjects analyzed using identical sequential steps. We found significant differences in metabolites between the three conditions. Principle component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) indicated clear grouping based on the metabolomes of the three conditions. We used machine learning algorithms and a percentage set of the data to train, and utilized a different or larger dataset to test whether a trained model can correctly classify the test dataset as PEX, POAG or control. Three different algorithms: linear support vector machines (SVM), deep learning, and a neural network were used for prediction. They all accurately classified the test datasets based on the AH metabolome of the sample. We next compared the AH metabolome with known AH and TM proteomes and genomes in order to understand metabolic pathways that may contribute to alterations in the AH metabolome in PEX. We found potential protein/gene pathways associated with observed significant metabolite changes in PEX.


Would older adults with mild cognitive impairment adhere to and benefit from a structured lifestyle activity intervention to enhance cognition?: a cluster randomized controlled trial.
Epidemiologic evidence suggests that cognitive and physical activities are associated with better cognition in late life. The present study was conducted to examine the possible benefits of four structured lifestyle activity interventions and compare their effectiveness in optimizing cognition for older adults with mild cognitive impairment (MCI). This was a 12-month cluster randomized controlled trial. 555 community-dwelling Chinese older adults with MCI (295 with multiple-domain deficits (mdMCI), 260 with single-domain deficit (sdMCI)) were recruited. Participants were randomized into physical exercise (P), cognitive activity (C), integrated cognitive and physical exercise (CP), and social activity (S, active control) groups. Interventions comprised of one-hour structured activities three times per week. Primary outcome was Clinical Dementia Rating sum of boxes (CDR-SOB) scores. Secondary outcomes included Chinese versions of Alzheimer's Disease Assessment Scale - Cognitive subscale (ADAS-Cog), delayed recall, Mini-Mental State Examination, Category Verbal Fluency Test (CVFT) and Disability Assessment for Dementia - Instrumental Activities of Daily Living (DAD-IADL). Percentage adherence to programs and factors affecting adherence were also examined. At 12th month, 423 (76.2%) completed final assessment. There was no change in CDR-SOB and DAD-IADL scores across time and intervention groups. Multilevel normal model and linear link function showed improvement in ADAS-Cog, delayed recall and CVFT with time (p<0.05). Post-hoc subgroup analyses showed that the CP group, compared with other intervention groups, had more significant improvements of ADAS-Cog, delayed recall and CVFT performance with sdMCI participants (p<0.05). Overall adherence rate was 73.3%. Improvements in ADAS-Cog and delayed recall scores were associated with adherence after controlling for age, education, and intervention groups (univariate analyses). Structured lifestyle activity interventions were not associated with changes in everyday functioning, albeit with some improvements in cognitive scores across time. Higher adherence was associated with greater improvement in cognitive scores. Factors to enhance adherence should be specially considered in the design of psychosocial interventions for older adults with cognitive decline. ClinicalTrials.gov ChiCTR-TRC-11001359.


Analysis of the amino acid sequences of plant Bowman-Birk inhibitors.
Plant seeds contain a large number of protease inhibitors of animal, fungal, and bacterial origin. One of the well-studied families of these inhibitors is the Bowman-Birk family(BBI). The BBIs from dicotyledonous seeds are 8K, double-headed proteins. In contrast, the 8K inhibitors from monocotyledonous seeds are single headed. Monocots also have a 16K, double-headed inhibitor. We have determined the primary structure of a Bowman-Birk inhibitor from a dicot, horsegram, by sequential edman analysis of the intact protein and peptides derived from enzymatic and chemical cleavage. The 76-residue-long inhibitor is very similar to that of Macrotyloma axillare. An analysis of this inhibitor along with 26 other Bowman-Birk inhibitor domains (MW 8K) available in the SWISSPROT databank revealed that the proteins from monocots and dicots belong to related but distinct families. Inhibitors from monocots show larger variation in sequence. Sequence comparison shows that a crucial disulphide which connects the amino and carboxy termini of the active site loop is lost in monocots. The loss of a reactive site in monocots seems to be correlated to this. However, it appears that this disulphide is not absolutely essential for retention of inhibitory function. Our analysis suggests that gene duplication leading to a 16K inhibitor in monocots has occurred, probably after the divergence of monocots and dicots, and also after the loss of second reactive site in monocots.


Redefining vaccination coverage and timeliness measures using electronic immunization registry data in low- and middle-income countries.
Vaccine coverage is routinely used as a performance indicator for immunization programs both at local and global levels. For many national immunization programs, there are challenges with accurately estimating vaccination coverage based on available data sources, however an increasing number of low- and middle-income countries (LMICs) have begun implementing electronic immunization registries to replace health facilities' paper-based tools and aggregate reporting systems. These systems allow for more efficient capture and use of routinely reported individual-level data that can be used to calculate dose-specific and cohort vaccination coverage, replacing the commonly used aggregate routine health information system data. With these individual-level data immunization programs have the opportunity to redefine performance measures to enhance programmatic decision-making at all levels of the health system. In this commentary, we discuss how measures for assessing vaccination status and program performance can be redefined and recalculated using these data when generated at the health facility level and the implications of the use and availability of electronic individual-level data.


KARS-related diseases: progressive leukoencephalopathy with brainstem and spinal cord calcifications as new phenotype and a review of literature.
KARS encodes lysyl- transfer ribonucleic acid (tRNA) synthetase, which catalyzes the aminoacylation of tRNA-Lys in the cytoplasm and mitochondria. Eleven families/sporadic patients and 16 different mutations in KARS have been reported to date. The associated clinical phenotype is heterogeneous ranging from early onset encephalopathy to isolated peripheral neuropathy or nonsyndromic hearing impairment. Recently additional presentations including leukoencephalopathy as predominant cerebral involvement or cardiomyopathy, isolated or associated with muscular and cerebral involvement, have been reported. A progressive Leukoencephalopathy with brainstem and spinal cord calcifications was previously described in a singleton patient and in two siblings, without the identification of the genetic cause. We reported here about a new severe phenotype associated with biallelic KARS mutations and sharing some common points with the other already reported phenotypes, but with a distinct clinical and neuroimaging picture. Review of KARS mutant patients published to date will be also discussed. Herein, we report the clinical, biochemical and molecular findings of 2 unreported Italian patients affected by developmental delay, acquired microcephaly, spastic tetraparesis, epilepsy, sensory-neural hypoacusia, visual impairment, microcytic hypochromic anaemia and signs of hepatic dysfunction. MRI pattern in our patients was characterized by progressive diffuse leukoencephalopathy and calcifications extending in cerebral, brainstem and cerebellar white matter, with spinal cord involvement. Genetic analysis performed on these 2 patients and in one subject previously described with similar MRI pattern revealed the presence of biallelic mutations in KARS in all 3 subjects. With our report we define the molecular basis of the previously described Leukoencephalopathy with Brainstem and Spinal cord Calcification widening the spectrum of KARS related disorders, particularly in childhood onset disease suggestive for mitochondrial impairment. The review of previous cases does not suggest a strict and univocal genotype/phenotype correlation for this highly heterogeneous entity. Moreover, our cases confirm the usefulness of search for common brain and spine MR imaging pattern and of broad genetic screening, in syndromes clinically resembling mitochondrial disorders in spite of normal biochemical assay.


To grow or not to grow: nutritional control of development during Caenorhabditis elegans L1 arrest.
It is widely appreciated that larvae of the nematode Caenorhabditis elegans arrest development by forming dauer larvae in response to multiple unfavorable environmental conditions. C. elegans larvae can also reversibly arrest development earlier, during the first larval stage (L1), in response to starvation. "L1 arrest" (also known as "L1 diapause") occurs without morphological modification but is accompanied by increased stress resistance. Caloric restriction and periodic fasting can extend adult lifespan, and developmental models are critical to understanding how the animal is buffered from fluctuations in nutrient availability, impacting lifespan. L1 arrest provides an opportunity to study nutritional control of development. Given its relevance to aging, diabetes, obesity and cancer, interest in L1 arrest is increasing, and signaling pathways and gene regulatory mechanisms controlling arrest and recovery have been characterized. Insulin-like signaling is a critical regulator, and it is modified by and acts through microRNAs. DAF-18/PTEN, AMP-activated kinase and fatty acid biosynthesis are also involved. The nervous system, epidermis, and intestine contribute systemically to regulation of arrest, but cell-autonomous signaling likely contributes to regulation in the germline. A relatively small number of genes affecting starvation survival during L1 arrest are known, and many of them also affect adult lifespan, reflecting a common genetic basis ripe for exploration. mRNA expression is well characterized during arrest, recovery, and normal L1 development, providing a metazoan model for nutritional control of gene expression. In particular, post-recruitment regulation of RNA polymerase II is under nutritional control, potentially contributing to a rapid and coordinated response to feeding. The phenomenology of L1 arrest will be reviewed, as well as regulation of developmental arrest and starvation survival by various signaling pathways and gene regulatory mechanisms.


Intestinal renin-angiotensin system is stimulated after deletion of Lkb1.
LKB1 is a serine-threonine kinase, mutation of which can lead to the development of multiple benign intestinal hamartomas (Peutz-Jeghers syndrome). In this study, the authors investigate the mechanisms underlying this phenotype by exploring the transcriptional changes associated with Lkb1 deletion in intestinal epithelium. The authors used mice with Lkb1 deleted in the intestinal epithelium using a Cyp1a1-specific inducible Cre recombinase and used Affymetrix (Santa Clara, California, USA) microarray analysis to examine the transcriptional changes occurring immediately after Lkb1 loss. The authors also generated crypt-villus organoid culture to analyse Lkb1 role in intestinal responses to exogenous stimuli. Affymetrix analysis identified the most significant change to be in Ren1 expression, a gene encoding a protease involved in angiotensinogen processing. Lkb1 deletion also enhanced ACE expression and subsequently angiotensin II (AngII) production in the mouse intestine. Intestinal apoptosis induced by Lkb1 deficiency was suppressed by ACE inhibitor captopril. Lkb1-deficient intestinal epithelium showed dynamic changes in AngII receptor type 1, suggesting a possible compensatory response to elevated AngII levels. A similar reduction in epithelial AngII receptor type 1 was also observed in human Peutz-Jeghers syndrome tumours contrasting with high expression of the receptor in the tumour stroma. Mechanistically, the authors showed two pieces of data that position Lkb1 in renin expression regulation, and they implied the importance of Lkb1 in linking cell responses with nutrient levels. First, the authors showed that Lkb1 deletion in isolated epithelial organoid culture resulted in renin upregulation only when the organoids were challenged with external cues such as AngII; second, that renin upregulation was dependent upon the MEK/ERK pathway in a circadian fashion and corresponded to active feeding time when nutrient levels were high. Taken together, these data reveal a novel role for Lkb1 in regulation of the gastrointestinal renin-angiotensin system.


Proteoglycan production is required in initial stages of new cartilage matrix formation but inhibits integrative cartilage repair.
The optimal stimulus to repair or regenerate cartilage is not known. We therefore modulated collagen deposition, collagen crosslinking and GAG deposition simultaneously during cartilage matrix production and integrative repair, creating more insight into their role in cartilage repair processes. Insulin-like growth factor 1 (IGF-1; increases proteoglycan and collagen synthesis), beta-aminopropionitrile (BAPN; a reversible inhibitor of collagen crosslinking) and para-nitrophenyl-beta-D-xyloside (PNPX; interferes with proteoglycan production) were used. Bovine articular chondrocytes were cultured in alginate beads for 3 weeks with or without IGF-1, BAPN or PNPX alone and in all possible combinations, followed by 3 weeks in control medium. DNA content, GAG and collagen deposition and collagen crosslinks were determined. Cartilage constructs were cultured under the same conditions and histologically analysed for integration of two opposing cartilage matrices. In alginate cultures, inhibition of collagen crosslinking with BAPN, in combination with promotion of matrix synthesis using IGF1, was most beneficial for matrix deposition. Addition of PNPX was always detrimental for matrix deposition. For integration of opposing cartilage constructs, the combination of BAPN, IGF1 and temporary prevention of proteoglycan formation with PNPX was most beneficial. When a new matrix is produced, proteoglycans are important to retain collagen in the matrix. When two already formed cartilage matrices have to integrate, a temporary absence of proteoglycans and temporary inhibition of collagen crosslinking might be more beneficial in combination with stimulation of collagen production, e.g. by IGF1. Therefore, the choice of soluble factors to promote cartilage regeneration depends on the type of therapy that will be used.


Conditioned medium from adipose-derived stem cells attenuates ischemia/reperfusion-induced cardiac injury through the microRNA-221/222/PUMA/ETS-1 pathway.
Rationale: Cardiovascular diseases, such as myocardial infarction (MI), are the leading causes of death worldwide. Reperfusion therapy is the common standard treatment for MI. However, myocardial ischemia/reperfusion (I/R) causes cardiomyocyte injury, including apoptosis and fibrosis. We aimed to investigate the effects of conditioned medium from adipose-derived stem cells (ADSC-CM) on apoptosis and fibrosis in I/R-treated hearts and hypoxia/reoxygenation (H/R)-treated cardiomyocytes and the underlying mechanisms. Methods: ADSC-CM was collected from ADSCs. The effects of intramuscular injection of ADSC-CM on cardiac function, cardiac apoptosis, and fibrosis examined by echocardiography, Evans blue/TTC staining, TUNEL assay, and Masson's trichrome staining in I/R-treated mice. We also examined the effects of ADSC-CM on apoptosis and fibrosis in H/R-treated H9c2 cells by annexin V/PI flow cytometry, TUNEL assay, and immunocytochemistry. Results: ADSC-CM treatment significantly reduced heart damage and fibrosis of I/R-treated mice and H/R-treated cardiomyocytes. In addition, the expression of apoptosis-related proteins, such as p53 upregulated modulator of apoptosis (PUMA), p-p53 and B-cell lymphoma 2 (BCL2), as well as the fibrosis-related proteins ETS-1, fibronectin and collagen 3, were significantly reduced by ADSC-CM treatment. Moreover, we demonstrated that ADSC-CM contains a large amount of miR-221/222, which can target and regulate PUMA or ETS-1 protein levels. Furthermore, the knockdown of PUMA and ETS-1 decreased the induction of apoptosis and fibrosis, respectively. MiR-221/222 overexpression achieved similar results. We also observed that cardiac I/R markedly increased apoptosis and fibrosis in miR-221/222 knockout (KO) mice, while ADSC-CM decreased these effects. The increased phosphorylation of p38 and NF-κB not only mediated myocardial apoptosis through the PUMA/p53/BCL2 pathway but also regulated fibrosis through the ETS-1/fibronectin/collagen 3 pathway. Conclusions: Overall, our results show that ADSC-CM attenuates cardiac apoptosis and fibrosis by reducing PUMA and ETS-1 expression, respectively. The protective effect is mediated via the miR-221/222/p38/NF-κB pathway.