Miracle-dz/UniFAR-FTUs huggingface dataset:
Structure of the human clamp loader reveals an autoinhibited conformation of a substrate-bound AAA+ switch

Significance DNA replication and repair depend on AAA+ ATPase protein complexes called clamp loaders that open and load ring-shaped sliding clamps onto DNA. Using cryogenic electron microscopy, we determined the first structure of the human clamp loader (RFC), which is in an autoinhibited conformation while bound to the sliding clamp (PCNA). We assign this to be a reaction intermediate prior to clamp opening and propose a conformational change necessary for activation, leading to a unique paradigm for the AAA+ ATPase mechanism. We examined RFC’s interaction with a PCNA disease variant, which illuminates how this variant maintains tight interactions with some partners. Finally, mapping of cancer mutations onto RFC’s structure suggests stability as a key factor in proper function and human health. DNA replication requires the sliding clamp, a ring-shaped protein complex that encircles DNA, where it acts as an essential cofactor for DNA polymerases and other proteins. The sliding clamp needs to be opened and installed onto DNA by a clamp loader ATPase of the AAA+ family. The human clamp loader replication factor C (RFC) and sliding clamp proliferating cell nuclear antigen (PCNA) are both essential and play critical roles in several diseases. Despite decades of study, no structure of human RFC has been resolved. Here, we report the structure of human RFC bound to PCNA by cryogenic electron microscopy to an overall resolution of ∼3.4 Å. The active sites of RFC are fully bound to adenosine 5′-triphosphate (ATP) analogs, which is expected to induce opening of the sliding clamp. However, we observe the complex in a conformation before PCNA opening, with the clamp loader ATPase modules forming an overtwisted spiral that is incapable of binding DNA or hydrolyzing ATP. The autoinhibited conformation observed here has many similarities to a previous yeast RFC:PCNA crystal structure, suggesting that eukaryotic clamp loaders adopt a similar autoinhibited state early on in clamp loading. Our results point to a “limited change/induced fit” mechanism in which the clamp first opens, followed by DNA binding, inducing opening of the loader to release autoinhibition. The proposed change from an overtwisted to an active conformation reveals an additional regulatory mechanism for AAA+ ATPases. Finally, our structural analysis of disease mutations leads to a mechanistic explanation for the role of RFC in human health.
Nonmetastatic, Castration-Resistant Prostate Cancer and Survival with Darolutamide.

BACKGROUND Darolutamide is a structurally distinct androgen-receptor inhibitor that is approved for the treatment of nonmetastatic, castration-resistant prostate cancer. In the planned primary analysis of a phase 3 trial, the median metastasis-free survival was significantly longer with darolutamide (40.4 months) than with placebo (18.4 months). The data for the analysis of overall survival were immature at the time of the primary analysis. METHODS In this double-blind, placebo-controlled trial, we randomly assigned 1509 men, in a 2:1 ratio, to receive darolutamide (955 patients) or placebo (554 patients) while they continued to receive androgen-deprivation therapy. After the results of the primary end-point analysis were found to be positive, unblinding of the treatment assignments occurred, and patients in the placebo group were permitted to cross over to receive open-label darolutamide treatment. At the time of this prespecified final analysis, which had been planned to be performed after approximately 240 deaths had occurred, overall survival and all other secondary end points were evaluated. RESULTS The median follow-up time was 29.0 months. At the time of unblinding of the data, all 170 patients who were still receiving placebo crossed over to receive darolutamide; 137 patients who had discontinued placebo before unblinding had occurred received at least one other life-prolonging therapy. Overall survival at 3 years was 83% (95% confidence interval [CI], 80 to 86) in the darolutamide group and 77% (95% CI, 72 to 81) in the placebo group. The risk of death was significantly lower, by 31%, in the darolutamide group than in the placebo group (hazard ratio for death, 0.69; 95% CI, 0.53 to 0.88; P = 0.003). Darolutamide was also associated with a significant benefit with respect to all other secondary end points, including the time to first symptomatic skeletal event and the time to first use of cytotoxic chemotherapy. The incidence of adverse events after the start of treatment was similar in the two groups; no new safety signals were observed. CONCLUSIONS Among men with nonmetastatic, castration-resistant prostate cancer, the percentage of patients who were alive at 3 years was significantly higher among those who received darolutamide than among those who received placebo. The incidence of adverse events was similar in the two groups. (Funded by Bayer HealthCare and Orion Pharma; ARAMIS ClinicalTrials.gov number, NCT02200614.).
Optimization of Conditions for the Production and Properties of Alginate-degrading Crude Enzyme from Shewanella oneidensis PKA 1008

An alginate-degrading bacterium, identified as Shewanella oneidensis PKA 1008 by 16S ribosomal RNA sequence analysis, was isolated from the green alga Ulva pertusa. Optimal conditions for the alginate-degrading ability of its crude enzyme were then determined. The optimal culture conditions for the growth of S. oneidensis PKA 1008 were pH 9, 2% NaCl, , and 24 hours incubation time. The crude enzyme produced by S. oneidensis PKA 1008 showed the highest alginate-degrading activity at pH 9, and produced 1.001 g of reducing sugar per liter in 3.5% (w/v) sodium alginate for 1 hour.
Implementation of perinatal collaborative care: a health services approach to perinatal depression care

Abstract Aim: Our objective was to integrate lessons learned from perinatal collaborative care programs across the United States, recognizing the diversity of practice settings and patient populations, to provide guidance on successful implementation. Background: Collaborative care is a health services delivery system that integrates behavioral health care into primary care. While efficacious, effectiveness requires rigorous attention to implementation to ensure adherence to the core evidence base. Methods: Implementation strategies are divided into three pragmatic stages: preparation, program launch, and program growth and sustainment; however, these steps are non-linear and dynamic. Findings: The discussion that follows is not meant to be prescriptive; rather, all implementation tasks should be thoughtfully tailored to the unique needs and setting of the obstetric community and patient population. In particular, we are aware that implementation on the level described here assumes commitment of both effort and money on the part of clinicians, administrators, and the health system, and that such financial resources are not always available. We conclude with synthesis of a survey of existing collaborative care programs to identify implementation practices of existing programs.
Three Rare Concurrent Complications of Tertiary Hyperparathyroidism: Maxillary Brown Tumor, Uremic Leontiasis Ossea, and Hungry Bone Syndrome

A 48-year-old woman in her 40’s with end-stage renal disease and tertiary hyperparathyroidism (HPT) presented for a rapidly progressive maxillary tumor. Initial workup was notable for elevated intact parathyroid hormone (PTH) and diffuse thickening of skull and facial bones on computed tomography, and maxillary tumor biopsy with multinucleated giant cells. She underwent subtotal parathyroidectomy (with removal of a parathyroid adenoma and 2 hyperplastic glands) and partial resection of maxillary brown tumor. The patient’s post-operative course was complicated by hungry bone syndrome, with hypocalcemia refractory to aggressive calcium repletion. Teriparatide (recombinant PTH) was utilized with rapid resolution of hypocalcemia. To our knowledge, this is the first case of maxillary brown tumor in tertiary HPT to be reported in the USA. This case also supports teriparatide as a novel therapeutic for hungry bone syndrome refractory to aggressive calcium repletion.
Anti-Inflammatory Activity and ROS Regulation Effect of Sinapaldehyde in LPS-Stimulated RAW 264.7 Macrophages

We evaluated the anti-inflammatory effects of SNAH in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages by performing nitric oxide (NO) assays, cytokine enzyme-linked immunosorbent assays, Western blotting, and real-time reverse transcription-polymerase chain reaction analysis. SNAH inhibited the production of NO (nitric oxide), reactive oxygen species (ROS), tumor necrosis factor (TNF)-α, and interleukin (IL)-6. Additionally, 100 μM SNAH significantly inhibited total NO and ROS inhibitory activity by 93% (p < 0.001) and 34% (p < 0.05), respectively. Protein expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) stimulated by LPS were also decreased by SNAH. Moreover, SNAH significantly (p < 0.001) downregulated the TNF-α, IL-6, and iNOS mRNA expression upon LPS stimulation. In addition, 3–100 µM SNAH was not cytotoxic. Docking simulations and enzyme inhibitory assays with COX-2 revealed binding scores of −6.4 kcal/mol (IC50 = 47.8 μM) with SNAH compared to −11.1 kcal/mol (IC50 = 0.45 μM) with celecoxib, a known selective COX-2 inhibitor. Our results demonstrate that SNAH exerts anti-inflammatory effects via suppression of ROS and NO by COX-2 inhibition. Thus, SNAH may be useful as a pharmacological agent for treating inflammation-related diseases.
Off-road assessment of cognitive fitness to drive

Abstract Road safety is a major issue in every society. The assessment of driving ability with a real vehicle is a lengthy and costly process; therefore, there is a growing need for the development of a neuropsychological battery that can provide a fast and reliable evaluation of a person’s cognitive fitness to drive. In the present study, we examined the relationship of an off-road lab-type test, namely, the Driving Scenes test, with performance on a driving simulator, as well as the influence of cognitive factors on driving ability as evaluated by Driving Scenes. Our results demonstrated a relationship between Driving Scenes and driving simulator performance. They also showed that some cognitive factors (namely, selective attention and verbal memory), were predictive of driving ability (as determined by the Driving Scenes test), but not others (namely visuospatial perception/memory, working memory, and visuospatial recognition). In addition, age strongly predicted performance on this test (younger age was associated with better performance). The conclusions derived from the present study highlight the need to identify off-road tools with high predictive value in assessing driving ability.
Behavioural and emotional profiles of children and adolescents with disorders of arousal

Disorders of arousals are common sleep disorders characterized by complex motor behaviours that arise episodically out of slow‐wave sleep. Psychological distress has long been associated with disorders of arousal, but this link remains controversial, especially in children and adolescents. The aim of this multi‐centre study was to characterize behavioural and emotional problems in a sample of children/adolescents with disorders of arousal, and to explore their relationship with the severity of nocturnal episodes. The parents of 41 children/adolescents with a diagnosis of disorders of arousal (11.5 ± 3.3 years old, 61% males) and of a group of 41 age‐ and gender‐matched control participants filled in the Child Behavior Checklist, along with the Sleep Disturbance Scale for Children and the Paris Arousal Disorders Severity Scale. Multilevel t‐tests revealed significantly higher total scores and sub‐scores of the Child Behavior Checklist for the patient group compared with the control group. Thirty‐four percent of the patients obtained pathological total scores, and 12% of them borderline scores. The severity of emotional/behavioural problems in the patient group was positively correlated with the severity of the nocturnal episodes. Interestingly, children/adolescents with disorders of arousal also obtained higher excessive daytime sleepiness and insomnia symptoms sub‐scores at the Sleep Disturbance Scale for Children. These results confirmed the hypothesis that behavioural/emotional problems are surprisingly common in children/adolescents with disorders of arousal. Further studies are warranted to investigate the causal relationship between pathological manifestations, subtler sleep abnormalities, and diurnal emotional/behavioural problems in children/adolescents with disorders of arousal.
Calcium intake from different food sources in Italian women without and with non-previously diagnosed osteoporosis

Abstract An adequate calcium and vitamin D intake may play a role in preventing osteoporosis, but the contribution of the different food sources of calcium with regards to the risk of osteoporosis been barely explored. This observational study evaluated the calcium intake through a food frequency questionnaire in 126 adult women with not previously diagnosed osteoporosis undergoing Dual-energy X-ray Absorptiometry (DXA) to screen for osteoporosis, and to correlate the calcium intake with parameters of bone density, measured by DXA. Total daily calcium intake and daily intake from food were similar among women found to have osteoporosis, osteopenia or normal condition. The main food source was milk and dairy products, while calcium supplementation was consumed by only 14% of subjects, irrespectively from osteoporosis conditions. DXA parameters were not significantly correlated with total daily calcium intake and calcium from food. The present study highlighted no qualitative and quantitative differences in the consumption of food groups contributing to calcium intakes in women with and without osteoporosis.
Social comparisons on social media: online appearance-related activity and body dissatisfaction in adolescent girls

Introduction: Adolescents’ engagement with online social networking platforms is advancing at an exponential rate and research is needed to investigate any impact on young users’ mental health. This study examined appearance-related activity (e.g. looking at photos of friends) on social media and body dissatisfaction among adolescent girls. Methods: Self-report measures of online appearance-related activity, social comparisons to female target groups, internalization of the thin ideal, body dissatisfaction, and self-esteem were administered to 210 girls (mean age = 15.16 years). Results: Body dissatisfaction was significantly related to (i) time spent engaged in social comparisons and (ii) upward social comparisons with various female targets while online. Evaluating oneself less favorably than the target group of close friends was most strongly associated with poorer body image appraisals. Serial multiple mediation analysis revealed that even after controlling for age and self-esteem, time spent engaged in social comparisons significantly mediated the relationship between online appearance-related activity and body dissatisfaction. This association was then further partially mediated by internalization of the thin ideal, which significantly mediated the relationship between time engaged in social comparisons and body dissatisfaction. Discussion: Results are discussed in terms of online social media platforms representing an additional appearance culture environment for adolescent girls. The effects of this on the mental health of vulnerable users and how future research should investigate protective factors that may buffer young girls from the adverse effects of social media are considered.
Effect of Low-Protein Diets with Crystalline Amino Acid Supplementation on Egg Production, Blood Parameters and Nitrogen Balance in Laying Japanese Quails

A total of 120 Japanese quails were randomly distributed into six dietary groups to evaluate the effect of low-crude protein (CP) diet supplemented with crystalline amino acids on productive and reproductive performance, blood parameters and nitrogen (N) balance. Treatments consisted of six different diets: a control diet (20% CP), a low-CP (16%) diet supplemented with threonine (Thr), valine (Val) and tryptophan (Trp), three low-CP diets supplemented with individual Thr, Val or Trp, and a low-CP diet supplemented with all aforementioned amino acids. None of the growth traits examined were influenced either by dietary treatments or by egg production (P > 0.05). The highest fertility percentages (88.2%) were observed in quails fed the low-CP diet without crystalline amino acids supplementation followed by low-CP diet supplemented with Val (86.2%), whereas the highest hatchability (from fertile eggs) percentage was observed in quails fed low-CP diet plus Trp. Egg shell percentages from quails fed the control diet were higher (P < 0.05) compared to other dietary treatments during the whole feeding period. Yolk percentage was decreased (P < 0.01) with the low-CP diet supplemented with amino acids. Blood protein and ammonia levels were decreased (P < 0.05) by reducing the dietary CP level or by supplementing amino acids to the low-CP diet. Reducing the CP level in the diet with amino acids supplementation in laying quails decreased the daily N excretion (P < 0.05). It is concluded that reducing the CP level in a diet supplemented with crystalline amino acids supports reproductive performance in quails, and is a valuable strategy for decreasing feeding cost and mitigating ammonia emission.
The road from cytotoxins to immunohistochemistry

ABSTRACT This review article traces the immunohistochemistry ancestry of cytotoxins as antibodies. The immunohistochemistry success, as a diagnostic and research test, stood on the shoulders of negative and positive experimentation results with cytotoxins from the first half of the twentieth century. This is when experimental immunologists came with the understanding of the need for both antigen and antibody purification to achieve specificity of the immunological reaction. Simultaneously, protocols were developed, which involved injecting antigenic material into experimental animals. During this time, reliable methods for evaluation of antiserum strength or titer were established. The evolution of antigen preparation for immunofluorescence is presented here as one of the transitional steps to modern immunohistochemistry. This work paved the way for the development of the monoclonal and polyclonal antibodies methodology. The article is written from the author’s perspective as an experimental immunologist and a hands-on participant during many steps of development in modern morphological laboratory diagnostics. Knowledge of the roots of immunohistochemistry is useful for laboratory professionals in appreciation of predecessors’ contributions. Familiarity with history of experimental immunology would be beneficial for understanding the methodological principles of their current work as well as future development prospects.
Chick Growth and Fledgling Performance of Streaked Shearwaters Calonectris leucomelas on Mikura Island for Two Breeding Seasons

Chick growth and fledgling performance of Streaked Shearwaters (n=89 in total) were studied on Mikura Island (33°52'N, 139°14'E), Izu Islands, Japan, in 1985 and 1990. Chicks hatched in mid August (13th, 17th) on average, with a mean body mass of 59-73g (11-13% of 552g mean adult mass). Adults stayed with their chicks for four days on average after hatching. Exposed culmen, tarsus, and wing lengths of chicks at hatching were 42%, 38% and 8% of mean adult sizes of 49, 51 and 310mm, respectively. Chicks attained mean adult body mass at around 40 days old in mid September. Peak body masses (734-739g) were reached on average in mid October at around 70 days old, and were about 33-34% heavier than mean adult size. Weight recession prior to fledging resulted in the loss of 30% of the peak body mass over the last three weeks, and chicks fledged at body masses of 519-522g that did not significantly differ from the 508g mean adult or subadult masses in autumn. Individual chicks increased body mass in an erratic fashion at intervals of from a few days to a week throughout the nestling period. Body mass and bony parts (culmen and tarsus) attained maximum increase rates (13.8g, 1.0mm and 0.6mm/day, respectively) by days 9-22, while flight feathers as well and tail feathers that emerged at 39 days on average, grew most rapidly at about 62-75 days (3.0mm/day [for remiges] and 4.3mm/day [for rectrices]). Chicks fledged almost at adult sizes (adult body mass and lengths of culmen and tail, but with 2% shorter tarsus and wing than adults). All of the growth rate constants ks, were very low at 0.05 for body mass, exposed culmen and wing, and at 0.08 for tarsus (only the tail constant was higher). These low values were typical of semi-precocial birds. Last feeding of chicks by parents occurred when chicks were 82 days old (7 November) on average and chicks were not fed for nine days on average prior to fledging. The mean length of the nestling period was 90-91 days, which ended on 13-17 November on average. Fledging success rate for chicks that had hatched was very low (21%) at the colony near the garbage dump, compared with two other areas with less human activity (68-72%). Most chicks were lost due to predation by feral cats and rats.
Expectations of Social Consequences Impact Anticipated Involvement in Health-Risk Behavior During Adolescence

This study examined how individual differences in expectations of social consequences relate to individuals’ expected involvement in health-risk behaviors (HRBs). A total of 122 adolescents (aged 11–17) reported their expected involvement in a number of risk behaviors and whether or not they expect to be liked more or less by engaging in the behavior: the expected social benefit. Higher perceived social benefit was associated with higher anticipated involvement in said behavior. This relationship was stronger for adolescents who reported a higher degree of peer victimization, supporting the hypothesis that experiencing victimization increases the social value of peer interactions. Findings suggest that adolescents incorporate expectations of social consequences when making decisions regarding their involvement in HRBs.