Antibody responses to Bacillus Calmette-Guérin during immunotherapy in bladder cancer patients.
Levels of serum antibody to Bacillus Calmette-Guérin (BCG) were determined by solid-phase radioimmunoassays in 48 normal donors and 60 patients with bladder cancer. Of 57 patients enrolled in a randomized prospective controlled trial of BCG immunotherapy, 47 were followed for up to 30 months, thus permitting comparisons of tumor recurrence, delayed cutaneous hypersensitivity responses to purified protein derivative (PPD), and serum BCG antibody levels at specific intervals during the clinical course. Sera from normal donors and cancer patients prior to BCG therapy had equally low levels of BCG antibody. AFter administration of intravesical and percutaneous BCG, significant rises of serum BCG antibody levels were detected in 23 of 24 randomized BCG immunotherapy patients. Skin test responses to PPD and serum BCG antibody levels had a close correlation as immune response indicators in 14 of 24 BCG therapy patients, while rises in serum BCG antibody levels were a better response indicator than PPD skin test reactions in the other 10 patients. Eleven of the 23 patients randomized into the non-BCG treatment group had tumor recurrence, although tumors recurred in only six of the 24 randomized patients in the BCG therapy group. Two additional nonrandomized BCG-treated patients had tumor recurrence. All eight BCG-treated patients with tumor recurrence had documented increases in serum BCG antibody levels after BCG therapy. Only three of these eight patients had conversion of PPD skin test responses from negative to positive; three were positive before immunotherapy and two remained negative after BCG therapy. Levels of antibodies reactive with human adenovirus type 5 and with Escherichia coli antigens were similar in sera from normal donors and from the randomized bladder cancer patients in both the BCG and non-BCG treatment groups. These results suggest that serum BCG antibody responses are as useful as PPD skin tests in identifying immunological responses to the immunoadjuvant BCG during immunotherapy trials in cancer patients.


It Happened to a Friend of Mine: The Influence of Perspective-taking on the Acknowledgment of Sexual Assault Following Ambiguous Sexual Encounters.
Failure to acknowledge that one has been the victim of sexual violence is an important, yet understudied, barrier that prevents women from seeking appropriate support following sexual violence. Drawing from a literature of demonstrating the benefits of self-distancing when evaluating emotionally charged personal information, the effects of self-distancing on acknowledgment of sexual assault were tested. Four experimental studies (Ntotal = 1,609) manipulated perspective-taking, either by asking women to imagine a series of hypothetical sexual encounters as experiences that happened to themselves or to their friends, or by asking women to describe a sexual experience from a first- or third-person perspective. Findings from the studies suggest that taking another person's perspective can help women to label ambiguous sexual experiences as more inappropriate and coercive. Notably, this did not seem to stem from women downplaying or dismissing experiences when they imagined themselves, as they reported anticipating more negative and less positive emotions in the scenarios where they imagined themselves compared to a friend. Nonetheless, in spite of the stronger anticipated negative emotional response when imagining themselves, women were less open to information about resources associated with sexual assault and support when they imagined themselves compared to a friend. This pattern of findings replicated for own, past sexual experiences but only to the extent that women spontaneously engaged in distanced perspective-taking themselves. This research suggests in addition to using contextual information to disambiguate and determine whether a sexual experience was inappropriate, taking a distanced perspective might provide a route through which women can come to terms with the experience and open up to the use of community-based services and sexual assault resources.


An improved Voronoi-diagram-based neural net for pattern classification.
We propose a novel two-layer neural network to answer a point query in R(n) which is partitioned into polyhedral regions; such a task solves among others nearest neighbor clustering. As in previous approaches to the problem, our design is based on the use of Voronoi diagrams. However, our approach results in substantial reduction of the number of neurons, completely eliminating the second layer, at the price of requiring only two additional clock steps. In addition, the design process is also simplified while retaining the main advantage of the approach, namely its ability to furnish precise values for the number of neurons and the connection weights necessitating neither trial and error type iterations nor ad hoc parameters.


Spinal epidural haematoma in haemophilia A.
We report on a 27-year-old male Caucasian with severe haemophilia A who presented with acute onset of neck pain with cervical nerve root irritation, due to a spinal epidural haematoma. His past medical history revealed carrying of a moderate weight as a possible traumatic mechanism. Under immediate factor VIII replacement therapy complete remission of the symptoms was achieved within several days. The diagnosis of spinal epidural haematoma and complete resorption was revealed by initial and follow-up magnetic resonance imaging studies of the cervical spine. Having reviewed the literature on spinal epidural haematoma, we present an overview of the treatment and outcome as regards haemophilia.


Surgical techniques for the repair of anterior mitral leaflet prolapse.
Myxomatous disease is the leading cause of mitral valve regurgitation in the developed world. Although posterior mitral leaflet (PML) prolapse is the most common cause of regurgitation and of repair, lesions of the anterior mitral leaflet (AML) are often considered beyond the possibilities of repair. The surgical anatomy and pathology of the AML and a precise terminology for the location of the lesions are described. The surgical maneuvers for commissural prolapse, chordal shortening, chordal replacement, leaflet resection, and annuloplasty are also described. The advantages and limitations of each maneuver are discussed. It is concluded that a flexible approach to the repair of the AML is necessary to adapt the best technique for each type of lesion. No particular technique can be applied systematically to all lesions. It is the author's contention that the majority of AML lesions can be repaired safely.


A linear, featured-data scheme for image reconstruction in time-domain fluorescence molecular tomography.
Fluorescence diffuse optical tomography (DOT) has attracted many attentions from the community of biomedical imaging, since it provides effective enhancement in imaging contrast. This modality is now rapidly evolving as a potential means of monitoring molecular events in small living organisms with help of molecule-specific contrast agents, referred to as fluorescence molecular tomography (FMT). FMT could greatly promote pathogenesis research, drug development, and therapeutic intervention. Although FMT in steady-state and frequency-domain modes have been heavily investigated, the extension to time-domain scheme is imminent for its several unique advantages over the others. By extending the previously developed generalized pulse spectrum technique for time-domain DOT, we propose a linear, featured-data image reconstruction algorithm for time-domain FMT that can simultaneously reconstruct both fluorescent yield and lifetime images of multiple fluorophores, and validate the methodology with simulated data.


Coresident and Noncoresident Emerging Adults' Daily Experiences With Parents.
Coresidence between emerging adults and parents is now common in the United States, but we know little about how coresidence influences daily experiences in these ties. Coresident (n = 62) and noncoresident (n = 97) emerging adults (aged 18-30) reported daily experiences with parents and mood for 7 days. During the study week, compared to offspring who lived apart from parents, coresident offspring were more likely to experience positive encounters, receive more support, wish parents would change, feel irritated, and report that their parents got on their nerves. Coresident offspring did not differ from noncoresident offspring with regard to stressful thoughts. Stressful thoughts about parents were associated with more negative daily mood; this effect did not differ for coresident and noncoresident offspring. Findings are discussed with regard to intergenerational ambivalence. In sum, coresident emerging adults were more involved with parents but not more affected by daily experiences with parents.


Allergen exposure chambers: harmonizing current concepts and projecting the needs for the future - an EAACI Position Paper.
Allergen exposure chambers (AECs) are clinical facilities allowing for controlled exposure of subjects to allergens in an enclosed environment. AECs have contributed towards characterizing the pathophysiology of respiratory allergic diseases and the pharmacological properties of new therapies. In addition, they are complementary to and offer some advantages over traditional multicentre field trials for evaluation of novel therapeutics. To date, AEC studies conducted have been monocentric and have followed protocols unique to each centre. Because there are technical differences among AECs, it may be necessary to define parameters to standardize the AECs so that studies may be extrapolated for driving basic immunological research and for marketing authorization purposes by regulatory authorities. For this task force initiative of the European Academy of Allergy and Clinical Immunology (EAACI), experts from academia and regulatory agencies met with chamber operators to list technical, clinical and regulatory unmet needs as well as the prerequisites for clinical validation. The latter covered the validation process, standardization of challenges and outcomes, intra- and interchamber variability and reproducibility, in addition to comparability with field trials and specifics of paediatric trials and regulatory issues. This EAACI Position Paper aims to harmonize current concepts in AECs and to project unmet needs with the intent to enhance progress towards use of these facilities in determining safety and efficacy of new therapeutics in the future.


Further characterization of rat liver mitochondrial fractions. Lipid composition and synthesis, and protein profiles.
1. Heavy and light mitochondrial fractions obtained by differential centrifugation were further characterized with respect to their lipid composition and synthesis and protein profiles, as seen by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. 2. The light mitochondrial fraction was rich in total lipids, phospholipids and cholesterol. The cardiolipin content, however, was low. 3. Rates of [3H]glycerol incorporation into phospholipids of heavy mitochondria and microsomal fractions were almost identical. On the other hand, incorporation into the individual phospholipids in light mitochondria was about 4-6 times higher. Incorporation into cardiolipin of light mitochondria was about 10-fold higher than in the heavy mitochondria. 4. Analysis of protein profiles by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis showed that the pattern obtained for the light mitochondria was similar to that for heavy mitochondria. However, the light fraction was relatively poor in high-molecular-weight proteins and rich in low-molecular-weight proteins. The microsomal protein profile was altogether different. 5. The significance of these findings is discussed in relation to mitochondrial biogenesis.


Non-infectious diseases in infectious disease consultation: A descriptive study in a tertiary care teaching hospital.
In this retrospective study, we aimed to investigate the frequency, trend, and nature of non-infectious diseases (non-IDs) as the final diagnosis for patients during an infectious disease (ID) consultation in an acute care hospital in Japan. This study included adult inpatients who underwent ID consultations between October 2016 and March 2018. The demographic data, clinical manifestations, and final non-ID diagnoses of cases were explored. Among the 502 patients who underwent ID consultations, 45 (9.0%) were diagnosed with non-IDs. The most common diagnoses were tumors (22.2%, n = 10), connective tissue and collagen vascular diseases (13.3%, n = 6), other inflammatory diseases (8.9%, n = 4), and drug-induced fever (8.9%, n = 4). Multiple logistic regression analysis showed that the presence of consultations for diagnosis (odds ratio [OR], 22.0; 95% confidence interval [CI], 10.1-48.2; p<0.01), consultations from the internal medicine department (OR, 2.5; 95% CI, 1.2-5.2; p = 0.02), and non-bacteremia cases (OR, 5.2; 95% CI, 1.4-19.3; p = 0.01) were independently associated with diagnosed non-IDs. Non-IDs after ID consultations were mainly tumor-related, inflammatory diseases, and drug fever. The presence of consultations for diagnosis, consultations from the internal medicine department and non-bacteremia cases were related to non-IDs among ID consultations. Further research is needed to explore the frequency and pattern of non-IDs to improve the quality of ID consultations in daily practice.


Lymphocytes bearing Fc receptors for IgE. IV. Formation of IgE-binding factor by rat T lymphocytes.
Culture of mesenteric lymph node cells from rats infected with Nippostrongylus brasiliensis resulted in the release of a soluble factor (IgE-binding factor) that can inhibit rosette formation of Fc epsilon R(+) lymphocytes with IgE-coated red cells. The factor is specifically absorbed with IgE-coated Sepharose. It has a m.w. of between 10,000 and 20,000. The major source of IgE-binding factor appears to be Fc epsilon R(+) T cells. The formation of IgE-binding factor by the lymphocytes was enhanced by IgE added to the culture medium. Evidence was obtained that Fc epsilon R(+) cells are involved in the induction of IgE-induced factor formation. Normal rat lymphocytes cultured alone failed to release IgE-binding factor, but incubation of normal lymphocytes with rat IgE resulted in the formation of IgE-binding factor and an increase in the proportion of Fc epsilon R(+) cells. It was also found that normal T cells formed the soluble factor upon incubation with IgE. In the induction of factor formation by normal lymphocytes, Fc gamma R(+) cells are essential; an Fc gamma R-depleted fraction failed to form IgE-binding factor upon incubation with IgE. The results suggest that interaction of IgE with Fc epsilon R(+) T cells and Fc gamma R(+) T cells induces the formation of IgE-binding factor(s).


Cannabis Legalization and Detection of Tetrahydrocannabinol in Injured Drivers.
The effect of cannabis legalization in Canada (in October 2018) on the prevalence of injured drivers testing positive for tetrahydrocannabinol (THC) is unclear. We studied drivers treated after a motor vehicle collision in four British Columbia trauma centers, with data from January 2013 through March 2020. We included moderately injured drivers (those whose condition warranted blood tests as part of clinical assessment) for whom excess blood remained after clinical testing was complete. Blood was analyzed at the provincial toxicology center. The primary outcomes were a THC level greater than 0, a THC level of at least 2 ng per milliliter (Canadian legal limit), and a THC level of at least 5 ng per milliliter. The secondary outcomes were a THC level of at least 2.5 ng per milliliter plus a blood alcohol level of at least 0.05%; a blood alcohol level greater than 0; and a blood alcohol level of at least 0.08%. We calculated the prevalence of all outcomes before and after legalization. We obtained adjusted prevalence ratios using log-binomial regression to model the association between substance prevalence and legalization after adjustment for relevant covariates. During the study period, 4339 drivers (3550 before legalization and 789 after legalization) met the inclusion criteria. Before legalization, a THC level greater than 0 was detected in 9.2% of drivers, a THC level of at least 2 ng per milliliter in 3.8%, and a THC level of at least 5 ng per milliliter in 1.1%. After legalization, the values were 17.9%, 8.6%, and 3.5%, respectively. After legalization, there was an increased prevalence of drivers with a THC level greater than 0 (adjusted prevalence ratio, 1.33; 95% confidence interval [CI], 1.05 to 1.68), a THC level of at least 2 ng per milliliter (adjusted prevalence ratio, 2.29; 95% CI, 1.52 to 3.45), and a THC level of at least 5 ng per milliliter (adjusted prevalence ratio, 2.05; 95% CI, 1.00 to 4.18). The largest increases in a THC level of at least 2 ng per milliliter were among drivers 50 years of age or older (adjusted prevalence ratio, 5.18; 95% CI, 2.49 to 10.78) and among male drivers (adjusted prevalence ratio, 2.44; 95% CI, 1.60 to 3.74). There were no significant changes in the prevalence of drivers testing positive for alcohol. After cannabis legalization, the prevalence of moderately injured drivers with a THC level of at least 2 ng per milliliter in participating British Columbia trauma centers more than doubled. The increase was largest among older drivers and male drivers. (Funded by the Canadian Institutes of Health Research.).


In Vivo Prevention of Implant-Associated Infections Caused by Antibiotic-Resistant Bacteria through Biofunctionalization of Additively Manufactured Porous Titanium.
Additively manufactured (AM) porous titanium implants may have an increased risk of implant-associated infection (IAI) due to their huge internal surfaces. However, the same surface, when biofunctionalized, can be used to prevent IAI. Here, we used a rat implant infection model to evaluate the biocompatibility and infection prevention performance of AM porous titanium against bioluminescent methicillin-resistant Staphylococcus aureus (MRSA). The specimens were biofunctionalized with Ag nanoparticles (NPs) using plasma electrolytic oxidation (PEO). Infection was initiated using either intramedullary injection in vivo or with in vitro inoculation of the implant prior to implantation. Nontreated (NT) implants were compared with PEO-treated implants with Ag NPs (PT-Ag), without Ag NPs (PT) and infection without an implant. After 7 days, the bacterial load and bone morphological changes were evaluated. When infection was initiated through in vivo injection, the presence of the implant did not enhance the infection, indicating that this technique may not assess the prevention but rather the treatment of IAIs. Following in vitro inoculation, the bacterial load on the implant and in the peri-implant bony tissue was reduced by over 90% for the PT-Ag implants compared to the PT and NT implants. All infected groups had enhanced osteomyelitis scores compared to the noninfected controls.


Measures of late effects in conservative treatment of breast cancer with standard or hypofractionated radiotherapy.
To confirm the equivalence in terms of late effects between two fractionation schedules of radiotherapy in conservative treatment of breast cancer. Fifty-eight patients treated at our institution from 1999 to 2002, with a median follow-up of 15 months (range, 7-46 months), were evaluated retrospectively. Twenty-nine patients (group A) were treated with standard fractionation: 5000 cGy/25fx/5 weeks, and 29 patients (group B) were treated with a hypofractionated schedule: 4500 cGy/15fx/5 weeks, three fractions per week. Late effects were evaluated using the LENT-SOMA scoring scale. The cosmetic results were assessed on a five-point scale. Skin elasticity was measured using a dedicated device (Cutometer SEM 575). There were no differences in breast volume, age at diagnosis and follow-up between groups. The LENT-SOMA toxicity observed in groups A and B, respectively, was as follows: grade 2-3 pain in five patients in each group; grade 2 breast edema in two and three patients; grade 2-3 and grade 2 fibrosis in six and eight patients; grade 2 and grade 2-3 telangiectasia in two and three patients; grade > or = 2 and 2 arm edema in two and one patients; no ulceration or atrophy were observed. Two patients in group A and one patient in group B needed treatment for breast and arm edema and arm edema, respectively. Very good, good-acceptable, and poor cosmetic results were observed in seven and two, fifteen and nineteen, and six and eight patients, respectively. Median skin elasticity loss due to treatment was -4.19% in group A and -6.29% in group B. These results are not statistically different. LENT-SOMA toxicities were minimal and no differences were observed between groups. Few patients in the hypofractionated group had very good cosmetic results, but it is debatable if radiotherapy was the only cause. Skin elasticity was not different between groups. Our results seem to suggest that it is possible to treat patients with both schedules, with similar late toxicity.


Correlation of foveal photoreceptor integrity with visual outcome in idiopathic epiretinal membrane.
To evaluate the correlation of foveal photoreceptor integrity with visual outcome in patients undergoing idiopathic epiretinal membrane (ERM) surgery using spectral domain optical coherence tomography (SD-OCT). Forty-three eyes of 43 patients were examined by SD-OCT before and at 1 and 6 months after vitrectomy for idiopathic ERM. Patients were divided into two groups based on the preoperative integrity of photoreceptor inner and outer segment junction (IS/OS) - Group I, intact IS/OS and Group D, disrupted IS/OS. Where present, disrupted length of IS/OS was measured, and the association between disrupted length of IS/OS and foveal thickness with best corrected visual acuity (BCVA) was evaluated. There were 27 eyes with an intact IS/OS (Group I) and 16 eyes with a disrupted IS/OS (Group D). Patients in Group I had significantly better postoperative BCVA and greater improvement than those in Group D at 6 months after surgery (p = 0.001 and p = 0.030, respectively). By 6 months after surgery, the disrupted length of IS/OS observed preoperatively in Group D had decreased significantly (p = 0.007). The disrupted length of IS/OS before and at 6 months after the surgery in Group D showed a positive correlation with BCVA (r = 0.632 and p = 0.009; r = 0.658 and p = 0.006, respectively). Foveal thickness did not correlate significantly with preoperative and postoperative BCVA. Preoperative disruption of foveal photoreceptors can be restored after ERM surgery. The preoperative integrity of foveal photoreceptors may be a clinically significant prognostic factor for visual recovery in patients with idiopathic ERM.


An efficient method for sonication assisted Agrobacterium-mediated transformation of coat protein (CP) coding genes into papaya (Carica papaya L.).
An efficient method for the production of transgenic papaya was developed via Sonication Assisted Agrobacterium-mediated Transformation (SAAT) of somatic embryos. The plasmid pGA482G was modified to contain gene PTi-Epj-TL-PLDMV with CP coding sequence of PLDMV Japan strain and chimeric gene PTi-NP-YKT with multiple CP coding sequences from PRSV Taiwan strain, PRSV Hawaii strain and PRSV Thailand strain, respectively. Disarmed Agrobacterium tumefaciens strain LBA4404 carrying the binary plasmid pGA482G with the CP genes and nptII gene was used to transform embryo calli of papaya variety Sunset to produce transgenic papaya plants. The experiment was focused on the screening of effective transformation method. The engineered Agrobacterium grown overnight was diluted with an infection media of high osmotic pressure (1/2 MS medium contain 6% sucrose and 1% glucose, pH 5.7) and adjusted to optical density OD600nm = 0.15-0.20, embryonic calli were immerged in it for 30 min and treated with 5 s, 15 s, and 20 s sonication respectively during the infection. Results indicated that 15 s sonication treatment improved the transformation efficiency dramatically. After 15 s sonication treatment on embryo calli loaded in 15 ml sterile plastic tubes, 21 putative transgenic lines were produced from 80 pieces embryonic calli (26.3%) transformed by Agrobacterium [pGA482G/CPG] and 8 putative transgenic lines was produced from 48 pieces embryonic calli (16.7%) transferred by Agrobacterium [pGA482G/CPB], while only a single line came out of 64 pieces embryonic calli (1.6%) transformed by Agrobacterium [pGA482G/CPG] and none from 25 pieces embryonic calli transformed by Agrobacterium [pGA482G/CPB] in the non-treatment control. Results also showed that the best concentration of selection antibiotic was 120 mg/L kanamycin. A total of 42 resistant shoots were produced from 421 pieces of original embryonic calli in 9 months. The presence of the CP genes in the transgenic plants and their integration into the papaya genome were confirmed by PCR and Southern hybridization respectively.


Inhibition of non-small cell lung cancer (NSCLC) proliferation through targeting G6PD.
Mounting evidence has linked cancer metabolic reprogramming with altered redox homeostasis. The pentose phosphate pathway (PPP) is one of the key metabolism-related pathways that has been enhanced to promote cancer growth. The glucose 6-phosphate dehydrogenase (G6PD) of this pathway generates reduced nicotinamide adenine dinucleotide phosphate (NADPH), which is essential for controlling cellular redox homeostasis. This research aimed to investigate the growth-promoting effects of G6PD in non-small cell lung cancer (NSCLC). Clinical characteristics and G6PD expression levels in lung tissues of 64 patients diagnosed with lung cancer at the King Chulalongkorn Memorial Hospital (Bangkok, Thailand) during 2009-2014 were analyzed. G6PD activity in NSCLC cell lines, including NCI-H1975 and NCI-H292, was experimentally inhibited using DHEA and siG6PD to study cancer cell proliferation and migration. The positive expression of G6PD in NSCLC tissues was detected by immunohistochemical staining and was found to be associated with squamous cells. G6PD expression levels and activity also coincided with the proliferation rate of NSCLC cell lines. Suppression of G6PD-induced apoptosis in NSCLC cell lines by increasing Bax/Bcl-2 ratio expression. The addition of D-(-)-ribose, which is an end-product of the PPP, increased the survival of G6PD-deficient NSCLC cell lines. Collectively, these findings demonstrated that G6PD might play an important role in the carcinogenesis of NSCLC. Inhibition of G6PD might provide a therapeutic strategy for the treatment of NSCLC.


Diversity in prevalence and characteristics of ESBL/pAmpC producing E. coli in food in Germany.
The spread of extended-spectrum β-lactamases (ESBLs) in Escherichia coli is a major public health issue and ESBL-producing bacteria are frequently reported in livestock. For the assessment of the role of the foodborne transmission pathway in Germany, detailed data on the prevalence and characteristics of isolates of food origin are necessary. The objective of this study was to describe the prevalence of cefotaxime resistant E. coli as well as ESBL/pAmpC-producing E. coli and their characteristics in foods in Germany. Out of 2256 food samples, the highest prevalence of cefotaxime resistant E. coli was observed in chicken meat (74.9%), followed by turkey meat (40.1%). Prevalence in beef, pork and minced meat was considerably lower (4.2-15.3%). Whereas 18.0% of the raw milk samples, collected at farm level were positive, this was true only for few cheese samples (1.3%). In one out of 399 vegetable samples a cefotaxime-resistant E. coli was isolated. ESBL resistance genes of the CTX-M-group (10.1% of all samples) were most frequently detected, followed by genes of the pAmpC (2.6%), SHV (2.0%) and TEM (0.8%) families. Distribution of ESBL/AmpC-encoding E. coli resistance genes and E. coli phylogroups was significantly different between the chicken related food samples and all other food items. Our study results reflect that consumers might get exposed to ESBL/pAmpC-producing E. coli through several food chains. These results together with those collected at primary production and in the human population in other studies will allow more detailed analysis of the foodborne pathways, considering transmission from livestock populations to food at retail and to consumers in Germany.


The influence of low-level laser therapy with alendronate irrigation on healing of bone defects in rats.
The aim of this study was to investigate the effects of alendronate (Aln) irrigation with low-level laser therapy (LLLT) on the healing of bone defects in rats. Sixty Wistar rats weighing 250 to 300 g were randomly divided into three groups of 20 animals each: (1) control group, (2) Aln group, and (3) Aln with LLLT group. The distal epiphysis of all rats was perforated with a surgical bone drill. Twenty rats served as control. The bone defects of 40 rats received local alendronate sodium trihydrate irrigation (1 mg/ml) at the time of surgery. LLLT was applied to the bone defects of 20 rats immediately after Aln irrigation, and repeated on days 2, 4, 6, and 8 with a total dose of 10 J/cm(2) (2 J/cm(2) × 5). Continuous wave of GaAlAs laser (808 nm) was used with a power density of 0.1 W/cm(2). Laser energy was applied for 20 s (0.1 W × 20 s/1 cm(2)) per session. Control group, Aln group, and Aln with LLLT group rats were sacrificed at days 10 and 20 to compare the bone healing of each group histologically. There were significant differences between the three groups regarding union, substantia spongiosa, cortex formation, and in sum of histologic scores on days 10 and 20 (P < 0.0001). Our findings demonstrated that Aln has a more positive effect with LLLT on bone healing in rats. It was concluded that combining LLLT (808 nm laser at 10 J/cm(2)) with Aln irrigation has a beneficial effect in bone repair. It was demonstrated experimentally that Aln irrigation during the surgery had a significant effect to enhance bone formation, and LLLT significantly potentiated the osseous healing effects of Aln on bone defects. This administration method is able to minimize the dose of Aln in order to avoid both systemic and local adverse effects as well as the local injection times during the bone healing process.


Nutritional regulation of insulin-like growth factor-I mRNA expression in barramundi, Lates calcarifer.
The effect of nutritional status on IGF-I mRNA expression in the liver and brain of juvenile barramundi (Lates calcarifer) was investigated. Fish were either fed a satiety ration (SAT) or starved (STV) for 6 weeks. Starved fish demonstrated significantly lower condition factor and hepatic IGF-I mRNA expression at 3 and 6 weeks, when compared with the SAT group. IGF-I mRNA expression in the brain was 10 fold lower than the liver and was not affected by ration size. These results suggest the liver is the major site of IGF-I mRNA synthesis and hepatic but not brain IGF-I mRNA expression is regulated by food availability in juvenile barramundi.


Time for a Systems Biological Approach to Cognitive Aging?-A Critical Review.
The underlying premise of current theories of cognitive decline with age tend to be primarily cognitive or biological explanations, with relatively few theories adequately integrating both aspects. Though literature has also emphasized the importance of several factors that contribute to cognitive aging including: (a) decline in sensory abilities; (b) the effect of motor speed on paper-pencil measures of cognitive speed; (c) the impact of level of education and physical activity; and (d) molecular biological changes that occur with age, these factors have seldom been implicated into any single theoretical model of cognitive aging. Indeed, such an integrated bio-cognitive model of aging has the potential to provide a more comprehensive understanding of attention, perception, learning, and memory across the lifespan. Thus, the aim of this review was to critically evaluate common theories of age-related cognitive decline and highlight the need for a more comprehensive systems neuroscience approach to cognitive aging.


Regulation of oocyte microvilli development in the baboon fetal ovary by estrogen.
We recently showed that the number of primordial follicles was reduced by 50% in ovaries of near-term fetal baboons deprived of estrogen in utero and restored to normal in animals supplemented with estrogen. Oocytes are avascular and rely on surrounding granulosa cells for nutrients, a process facilitated by microvilli on the oocyte surface. However, our understanding of oocyte microvillus development in the primate fetal ovary is incomplete. Thus, we determined whether estrogen regulates formation of oocyte microvilli in utero. Fetal ovaries were obtained on d 165 gestation (term = d 184) from baboons untreated (n = 3) or treated on d 100-165 with aromatase inhibitor CGS 20267 (estrogen suppressed by 95%; n = 5) or CGS 20267 and estradiol (n = 4). Follicles with intact (homogeneous cytoplasm) or nonintact (cytoplasm vacuolated) oocytes were quantified and the number/height of oocyte microvilli determined by electron microscopy. In untreated baboons, the mean (+/-se) number of follicles/0.08 mm(2) with an intact oocyte (11.5 +/- 0.5) was decreased (P < 0.05) by 70% in fetal ovaries of estrogen-suppressed baboons (3.4 +/- 0.2) and restored (P < 0.05) by CGS 20267 and estradiol (11.2 +/- 1.2). In estrogen-deprived fetuses, the number of microvilli/intact oocyte (23 +/- 3) was 56% lower (P < 0.01) than normal (52 +/- 5) and restored by CGS 20267 and estrogen (62 +/- 4). Moreover, in intact oocytes of estrogen-suppressed baboons, height (nm) of microvilli (105 +/- 11) was 54-62% lower (P < 0.01) than in intact oocytes of fetal ovaries of untreated (228 +/- 13) or estrogen-treated (274 +/- 17) baboons. In estrogen-replete baboons, the number of microvilli in intact oocytes was 2-fold greater (P < 0.01) than in nonintact oocytes. However, in estrogen-deprived baboons, no microvilli were detected in nonintact oocytes and the number of microvilli in intact oocytes was similar to that in nonintact oocytes of untreated fetuses. We conclude that development of microvilli in oocytes of primordial follicles in the primate fetal ovary is regulated by estrogen. Collectively, these results and those of our previous studies indicate that estrogen regulates fetal ovarian folliculogenesis and development of follicles with oocytes composed of microvilli critical for nutrient uptake and presumably long-term survival.


Barriers and facilitators related to mental health care use among older veterans in the United States.
Psychiatric disorders are more prevalent among older veterans compared with their civilian counterparts, but many veterans with symptoms of psychiatric disorders do not utilize mental health services. This study examined barriers and facilitators related to current mental health care utilization in a nationally representative sample of veterans ages 60 and older (N=2,025). Using data from the National Health and Resilience in Veterans Study, the authors evaluated how predisposing, enabling, and need characteristics as well as perceived barriers to care were related to utilization of mental health care among older veterans. A minority of veterans (N=130; weighted prevalence, 6%) reported current mental health care utilization. Among veterans (N=144) who screened positive for a current psychiatric disorder, 42 (weighted prevalence, 25%) were currently utilizing services. In the full sample, current utilization was associated with lifetime posttraumatic stress disorder or depression (odds ratio [OR]=5.88, 95% confidence interval [CI]=3.51-9.84), lifetime drug use disorder (OR=2.87, CI=1.59-5.17), severity of current psychiatric symptoms (OR=1.40, CI=1.19-1.65), general medical difficulties (OR=1.28, CI=1.10-1.50), and lower perceptions of stigma (OR=.80, CI=.68-.93). Non-Hispanic veterans were less likely to utilize care (OR=.42, CI=.25-.69). Among psychiatrically distressed veterans, current utilization was associated with younger age (OR=.89, CI=.81-.97), current suicidal ideation (OR=5.60, CI=1.98-15.84), and fewer negative beliefs about mental health care (OR=.23, CI=.09-.56). Efforts to identify psychiatrically distressed veterans and to reduce stigma and negative beliefs about mental health care may help increase mental health service utilization among older U.S. veterans.


Priority interventions to improve maternal and child diets in Sub-Saharan Africa and South Asia.
Nutrition-sensitive interventions to improve overall diet quality are increasingly needed to improve maternal and child health. This study demonstrates feasibility of a structured process to leverage local expertise in formulating programmes tailored for current circumstances in South Asia and Africa. We assembled 41 stakeholders in 2 regional workshops and followed a prespecified protocol to elicit programme designs listing the human and other resources required, the intervention's mechanism for impact on diets, target foods and nutrients, target populations, and contact information for partners needed to implement the desired programme. Via this protocol, participants described 48 distinct interventions, which we then compared against international recommendations and global goals. Local stakeholders' priorities focused on postharvest food systems to improve access to nutrient-dense products (75% of the 48 programmes) and on production of animal sourced foods (58%), as well as education and social marketing (23%) and direct transfers to meet food needs (12.5%). Each programme included an average of 3.2 distinct elements aligned with those recommended by United Nations system agencies in the Framework for Action produced by the Second International Conference on Nutrition in 2014 and the Compendium of Actions for Nutrition developed for the Renewed Efforts Against Child Hunger initiative in 2016. Our results demonstrate that a participatory process can help local experts identify their own priorities for future investments, as a first step in a novel process of rigorous, transparent, and independent priority setting to improve diets among those at greatest risk of undernutrition.


Effects of feeding a high- or moderate-starch prepartum diet to cows on newborn dairy heifer calf responses to intravenous glucose tolerance tests early in life.
The objective of this study was to evaluate the effect of feeding a prepartum diet with a high or moderate starch content on growth and insulin sensitivity of female offspring early in life. Thirty-eight Holstein heifer calves were born to dams fed either a high-starch (26% starch on a DM basis, HI; n = 20) or moderate-starch (14% starch on a DM basis, MOD; n = 18) prepartum diet commencing at 28 ± 3 d before expected parturition date. Following birth, all calves were housed individually and fed three 2-L meals of colostrum within the first 24 h of life and offered 10 L/d of milk replacer (26% CP, 18% fat, mixed to 130 g/L). Body weight of calves was measured at birth and on d 2 (after colostrum feeding but before milk feeding), 10 ± 2, and 20 ± 2. A glucose tolerance test was performed at a minimum of 6 h after their last colostrum or milk meal to evaluate insulin sensitivity on d 2, 10 ± 2 and 20 ± 2. Body weight did not differ throughout between HI and MOD calves; however, calves born to primiparous dams were smaller compared with those born to multiparous dams. Glucose or insulin concentrations were not different before the glucose tolerance test. Following the glucose tolerance test, maximum glucose concentrations were not different between treatments at any time point. However, HI calves had greater insulin area under the curve, and HI calves had greater maximum insulin concentrations on d 2. Glucose or insulin clearance rates were not different nor was the calculated insulin sensitivity index between treatments. These findings suggest that feeding a HI prepartum diet may reduce some insulin sensitivity indicators of female offspring early in life.


The reaction of human alpha 2-macroglobulin with alpha-chymotrypsin. A stopped-flow kinetic investigation.
The kinetics of the conformational changes of human alpha 2-macroglobulin (alpha 2M) induced by reaction with pure alpha-chymotrypsin, have been analyzed using three fluorescent probes, namely protein tryptophan groups and the dye 6-(4-toluidino)-2-naphthalenesulfonate, to monitor alterations of the alpha 2M structure, and a covalent conjugate of chymotrypsin and fluorescein isothiocyanate (Chy-FITC). The main reaction sequence exhibits a triphasic time course with any of the labels used. Each phase is first-order. The fixation of a single molecule of chymotrypsin to one protease-binding site of alpha 2M (site A) initiates the whole process and determines the access to the second site (site B). Of the three exponential phases of the reaction (20 degrees C), phase I (k1 approximately 19.6 min-1) and phase II (k2 approximately 5.3 min-1) belong to site A. Phase III is related to site B transformation. It contains two steps with different responses from tryptophan (k3 approximately 0.77 min-1) and Chy-FITC (k3 approximately 0.19 min-1) fluorescence measurements. The point to be stressed is that site A and site B, while presumably identical in the native form, are not equivalent with regard to their fluorescence and kinetic properties. However, the activation energy (E = 30.1 +/- 2.7 kJ mol-1) is the same for the three phases of the reaction. When present in sufficient excess, free chymotrypsin or native alpha 2M is able to form reversible complexes with the above-related chymotrypsin-alpha 2M adducts. Only the alpha 2M site A core seems to be involved in this parallel process. In addition the conformational state of the chymotrypsin-alpha 2M complexes is shown to depend on the pH, with a pKa of 6.4.


Biomarkers of pancreatic cancer.
Pancreatic cancer has a high mortality rate since early diagnosis is difficult and radical operation is challenging. Classical tumor markers are reliable parameters to determine disease progression during chemotherapy or recurrence after surgery, but they are not adequate to identify suspected disease or for screening. Endoscopic brushing cytology or biopsy from the stenotic duct is widely performed for the histological evidence of pancreatic cancer, but still suffers from low sensitivity. Recently, several molecules were found to be specifically expressed in pancreatic cancer, and these novel molecular markers are reported to improve the sensitivity of cytology or biopsy. In some cases, novel markers are tested for the diagnosis of cystic neoplasms. In addition, advances in endoscopic ultrasonography-guided fine needle aspiration biopsy enabled sampling of the cancer tissue before surgery or treatment, which delineates the individualized therapeutic strategy against pancreatic cancer, via the assessment of prognosis- or therapy resistance-related factors. Furthermore, novel transcriptomic or metabolomic biomarkers in the clinical samples collected by non-invasive methods, e.g. blood or saliva samples, are now applied for the diagnosis of pancreatic cancer. These methods will be beneficial for the screening and early detection of pancreatic cancer.


Trypanosoma cruzi Induces B Cells That Regulate the CD4+ T Cell Response.
Trypanosoma cruzi infection induces a polyclonal B cell proliferative response characterized by maturation to plasma cells, excessive generation of germinal centers, and secretion of parasite-unrelated antibodies. Although traditionally reduced to the humoral response, several infectious and non-infectious models revealed that B lymphocytes could regulate and play crucial roles in cellular responses. Here, we analyze the trypomastigote-induced effect on B cells, their effects on CD4+ T cells, and their correlation with in vivo findings. The trypomastigotes were able to induce the proliferation and the production of IL-10 or IL-6 of naïve B cells in co-culture experiments. Also, we found that IL-10-producing B220lo cells were elicited in vivo. We also found up-regulated expression of FasL and PD-L1, proteins involved in apoptosis induction and inhibition of TCR signaling, and of BAFF and APRIL mRNAs, two B-cell growth factors. Interestingly, it was observed that IL-21, which plays a critical role in regulatory B cell differentiation, was significantly increased in B220+/IL-21+ in in vivo infections. This is striking since the secretion of IL-21 is associated with T helper follicular cells. Furthermore, trypomastigote-stimulated B-cell conditioned medium dramatically reduced the proliferation and increased the apoptotic rate on CD3/CD28 activated CD4+ T cells, suggesting the development of effective regulatory B cells. In this condition, CD4+ T cells showed a marked decrease in proliferation and viability with marginal IL-2 or IFNγ secretion, which is counterproductive with an efficient immune response against T. cruzi. Altogether, our results show that B lymphocytes stimulated with trypomastigotes adopt a particular phenotype that exerts a strong regulation of this T cell compartment by inducing apoptosis, arresting cell division, and affecting the developing of a proinflammatory response.


Characterization of a high-molecular-weight Notch complex in the nucleus of Notch(ic)-transformed RKE cells and in a human T-cell leukemia cell line.
Notch genes encode a family of transmembrane proteins that are involved in many cellular processes, such as differentiation, proliferation, and apoptosis. It is well established that all four Notch genes can act as oncogenes; however, the mechanism by which Notch proteins transform cells remains unknown. Previously, we reported that both nuclear localization and transcriptional activation are required for neoplastic transformation of RKE cells. Furthermore, we identified cyclin D1 as a direct transcriptional target of constitutively active Notch molecules. In an effort to understand the mechanism by which Notch functions in the nucleus, we sought to determine if Notch formed stable complexes using size exclusion chromatography. Herein, we report that the Notch intracellular domain (N(ic)) forms distinct high-molecular-weight complexes in the nuclei of transformed RKE cells. The largest complex is approximately 1.5 MDa and contains both endogenous CSL (for CBF1, Suppressor of Hairless, and Lag-1) and Mastermind-Like-1 (Maml). N(ic) molecules that do not have the high-affinity binding site for CSL (RAM) retain the ability to associate with CSL in a stable complex through interactions involving Maml. However, Maml does not directly bind to CSL. Furthermore, Maml can rescue Delta RAM transcriptional activity on a CSL-dependent promoter. These results indicate that deletion of the RAM domain does not equate to CSL-independent signaling. Moreover, in SUP-T1 cells, N(ic) exists exclusively in the largest N(ic)-containing complex. SUP-T1 cells are derived from a T-cell leukemia that harbors the t(7;9)(q34;q34.3) translocation and constitutively express N(ic). Taken together, our data indicate that complex formation is likely required for neoplastic transformation by Notch(ic).


Improving care of migrants is key for viral hepatitis elimination in Europe.
By 2040, deaths from chronic viral hepatitis worldwide are projected to exceed those from human immunodeficiency virus infection, tuberculosis and malaria combined. The burden of this disease is predominantly carried by low-resource countries in Africa and Asia. In resource-rich countries, the epidemiological spread of viral hepatitis is partially driven by migrant movements from areas of high endemicity. In the last decade, Member States of the European Union and the European Economic Area have experienced an unprecedented influx of migrants, which has resulted in the polarization of political views about migration. In addition, the coronavirus disease 2019 pandemic has worsened the economic and health conditions of migrants and contributed to hostility to ensuring their health rights. Moreover, the implementation of hostile laws in some host nations has increased the vulnerability of marginalized migrant subgroups, such as asylum seekers and undocumented individuals. These developments have complicated the historical challenge of identifying high-risk migrant groups for screening and treatment. However, if European countries can apply the simplified assessment tools and diagnostic tests for viral hepatitis that have been used for decentralized screening and monitoring in resource-poor countries, the uptake of care by migrants could be dramatically increased. Given the global calls for the elimination of viral hepatitis, European nations should recognize the importance of treating this vulnerable migrant population. Political and health strategies need to be adapted to meet this challenge and help eliminate viral hepatitis globally.


Phosphatidylinositol-dependent membrane fusion induced by a putative fusogenic sequence of Ebola virus.
The membrane-interacting abilities of three sequences representing the putative fusogenic subdomain of the Ebola virus transmembrane protein have been investigated. In the presence of calcium, the sequence EBO(GE) (GAAIGLAWIPYFGPAAE) efficiently fused unilamellar vesicles composed of phosphatidylcholine, phosphatidylethanolamine, cholesterol, and phosphatidylinositol (molar ratio, 2:1:1:0.5), a mixture that roughly resembles the lipid composition of the hepatocyte plasma membrane. Analysis of the lipid dependence of the process demonstrated that the fusion activity of EBO(GE) was promoted by phosphatidylinositol but not by other acidic phospholipids. In comparison, EBO(EA) (EGAAIGLAWIPYFGPAA) and EBO(EE) (EGAAIGLAWIPYFGPAAE) sequences, which are similar to EBO(GE) except that they bear the negatively charged glutamate residue at the N terminus and at both the N and C termini, respectively, induced fusion to a lesser extent. As revealed by binding experiments, the glutamate residue at the N terminus severely impaired peptide-vesicle interaction. In addition, the fusion-competent EBO(GE) sequence did not associate significantly with vesicles lacking phosphatidylinositol. Tryptophan fluorescence quenching by vesicles containing brominated phospholipids indicated that the EBO(GE) peptide penetrated to the acyl chain level only when the membranes contained phosphatidylinositol. We conclude that binding and further penetration of the Ebola virus putative fusion peptide into membranes might be governed by the nature of the N-terminal residue and by the presence of phosphatidylinositol in the target membrane. Moreover, since insertion of such a peptide leads to membrane destabilization and fusion, the present data would be compatible with the involvement of this sequence in Ebola virus fusion.


Topical and oral ketorolac administration increases the intraocular pressure-lowering effect of latanoprost.
To verify the influence of a non-steroidal anti-inflammatory drug (NSAID), ketorolac (topical and oral) on the intraocular pressure reduction induced by 0.005% latanoprost topical administration, both in patients affected by primary open-angle glaucoma and in healthy controls. Two groups of subjects were enrolled for this randomized, prospective, masked clinical study: 16 glaucomatous patients well controlled with 0.005% latanoprost eyedrops (group I) and 16 healthy adult volunteers (group II). Group I subjects were treated at one-week intervals with 10 mg of oral ketorolac, oral placebo, topical ketorolac, and topical placebo, respectively; for each administration modality, the switch between drug and placebo was performed in a randomized, crossover, double-blind fashion. Group II subjects followed the same protocol, with the topical once-daily 0.005% latanoprost treatment starting three days prior to the ketorolac/placebo administration. Intraocular pressure (IOP) was investigated in both groups on the day of oral/topical administration of ketorolac or placebo at baseline (8:00 AM) and at the following intervals: 1, 2, 4, 8, 12, and 24 hours. No significant IOP changes after oral and topical placebo administration were observed in either group. In contrast, when the subjects received ketorolac (either oral or topical), a marked decrease in IOP was recorded, with a noticeable fall at the first hour after the NSAID administration (p = 0.01), which remained still significant 8 hours later (p < 0.05). Topical and oral ketorolac strengthens the latanoprost-induced IOP-lowering effect both in glaucomatous patients and in healthy subjects.


L-ferritin binding to scara5: a new iron traffic pathway potentially implicated in retinopathy.
Iron is essential in the retina because the heme-containing enzyme guanylate cyclase modulates phototransduction in rods and cones. Transferrin endocytosis is the classical pathway for obtaining iron from the blood circulation in the retina. However, the iron storage protein ferritin has been also recently proposed as an iron carrier. In this study, the presence of Scara5 and its binding to L-ferritin was investigated in the retina. Our results showed that Scara5, the specific receptor for L-ferritin, was expressed in mouse and human retinas in many cell types, including endothelial cells. Furthermore, we showed that intravenously injected ferritin crossed the blood retinal barrier through L-ferritin binding to Scara5 in endothelial cells. Thus, suggesting the existence of a new pathway for iron delivery and trafficking in the retina. In a murine model of photoreceptor degeneration, Scara5 was downregulated, pointing out this receptor as a potential player implicated in retinopathy and also as a possible therapeutic target.


Femur fractures in professional athletes: a case series.
To discuss return to play after femur fractures in several professional athletes. Femur fractures are rare injuries and can be associated with significant morbidity and mortality. No reports exist, to our knowledge, on return to play after treatment of isolated femur fractures in professional athletes. Return to play is expected in patients with femur fractures, but recovery can take more than 1 year, with an expected decrease in performance. Four professional athletes sustained isolated femur fractures during regular-season games. Two athletes played hockey, 1 played football, and 1 played baseball. Three players were treated with anterograde intramedullary nails, and 1 was treated with retrograde nailing. All players missed the remainder of the season. At an average of 9.5 months (range, 7-13 months) from the time of injury, all athletes were able to return to play. One player required the removal of painful hardware, which delayed his return to sport. Final radiographs revealed that all fractures were well healed. No athletes had subjective complaints or concerns that performance was affected by the injury at an average final follow-up of 25 months (range, 22-29 months). As the size and speed of players increase, on-field trauma may result in significant injury. All players returned to previous levels of performance or exceeded previous statistical performance levels. In professional athletes, return to play from isolated femur fractures treated with either an anterograde or retrograde intramedullary nail is possible within 1 year. Return to the previous level of performance is possible, and it is important to develop management protocols, including rehabilitation guidelines, for such injuries. However, return to play may be delayed by subsequent procedures, including hardware removal.


A Randomized Controlled Clinical Trial Comparing Conventional and Computer-Assisted Implant Planning and Placement in Partially Edentulous Patients. Part 1: Clinician-Related Outcome Measures.
The objectives of this study were to compare conventional and computer-assisted implant planning and placement (CAIPP) protocols regarding surgical planning predictability, intraoperative complications, and patient-centered outcomes. Partially edentulous patients (N = 73) were randomly allocated to one of three treatment groups: control (C, n = 26), with preoperative planning based on conventional radiography and freehand implant placement; and test 1 (T1, n = 24) and test 2 (T2, n = 23), with two different CAIPP protocols. The clinicians' predictions of the bony morphology, materials needed for surgery, and surgery duration were matched with intrasurgical findings using kappa tests. Complications or deviations from the surgical or prosthetic protocol were recorded. Descriptive statistics were used to study the sample sorted out by treatment group. Differences between groups were evaluated with chi-square test for qualitative variables and with nonparametric Kruskal-Wallis test for quantitative continuous variables. For post-hoc tests, the Bonferroni corrected (P < .016 = .05/3) Mann-Whitney test was used. CAIPP protocols showed better diagnostic potential than conventional protocols for the bone topography, need for simultaneous GBR procedures, membrane selection, and implant length predictions. The rate of surgical protocol deviations was similar in all groups, but their nature differed. Conventional protocols showed fewer splint-related incidences. Implant bed preparation and insertion could not be fully completed using the surgical splint in 3.8% of patients in C (1/26), 45.8% in T1 (11/24), and 47.8% in T2 (11/23). Deviation from the initial prosthetic plan was necessary in one case (T2; 4.4%). No biologic or technical complications were observed. CAIPP protocols showed a higher diagnostic potential than conventional protocols. A high incidence of intraoperative surgical protocol modifications to adjust suboptimal implant placements was reported for every group. Therefore, strict intraoperative implant position monitoring is mandatory for both treatment protocols.


Occurrence of Naegleria fowleri and faecal indicators in sediments from Lake Pontchartrain, Louisiana.
The occurrence of amoeba, Naegleria fowleri, in sediment samples from Lake Pontchartrain in Louisiana was investigated. This amoeba is pathogenic and can cause primary amoebic meningoencephalitis. In this study, quantitative polymerase chain reaction methods were used to test for the prevalence of Naegleria fowleri, HF183, and E. coli. N. fowleri was detected in 51.25% of our sediment samples. Illumina sequencing of sediment samples revealed ten different phyla, with Cyanobacteria being the most predominant at sites that generally presented with the highest median N. fowleri concentrations. N. fowleri was however strongly negatively correlated with HF183 (r = -0.859, p < 0.001). Whenever sediment E. coli concentrations were below 1.54 Log GC/g, there was only a 37.5% chance that N. fowleri would be detected in the same sample. When sediment E. coli concentrations exceeded 2.77 Log GC/g, the chances of detecting N. fowleri in the same sample increased to 90%, potentially suggesting predatory activity by the amoeba. The effect of temperature was observed to be different in relation to observed N. fowleri concentrations and detection rates. Although sediment samples collected during periods of higher temperatures had significantly lower mean N. fowleri concentrations (2.7 Log GC/g) compared to those collected at lower temperatures (3.7 Log GC/g, t(39) = 4.167, p < 0.001), higher N. fowleri detection rates in the overall samples were observed at higher temperatures (>19.1 °C) than at lower temperatures (<19.1 °C).


Salivary DNA Loads for Human Herpesviruses 6 and 7 Are Correlated With Disease Phenotype in Myalgic Encephalomyelitis/Chronic Fatigue Syndrome.
Myalgic Encephalomyelitis/Chronic Fatigue Syndrome (ME/CFS) is a complex chronic condition affecting multiple body systems, with unknown cause, unclear pathogenesis mechanisms, and fluctuating symptoms which may lead to severe debilitation. It is frequently reported to have been triggered by an infection, but there are no clear differences in exposure to, or seroprevalence of, any particular viruses between people with ME/CFS and healthy individuals. However, herpes viruses have been repeatedly hypothesized to underlie the chronic relapsing/remitting form of MS/CFS due to their persistence in a latent form with periodic reactivation. It is possible that ME/CFS is associated with herpes virus reactivation, which has not been detectable previously due to insufficiently sensitive testing methods. Saliva samples were collected from 30 people living with ME/CFS at monthly intervals for 6 months and at times when they experienced symptom exacerbation, as well as from 14 healthy control individuals. The viral DNA load of the nine humanherpes viruses was determined by digital droplet PCR. Symptoms were assessed by questionnaire at each time point. Human herpesvirus (HHV) 6B, HHV-7, herpes simplex virus 1 and Epstein-Barr virus were detectable within the saliva samples, with higher HHV-6B and HHV-7 viral loads detected in people with ME/CFS than in healthy controls. Participants with ME/CFS could be broadly separated into two groups: one group displayed fluctuating patterns of herpesviruses detectable across the 6 months while the second group displayed more stable viral presentation. In the first group, there was positive correlation between HHV-6B and HHV-7 viral load and severity of symptom scores, including pain, neurocognition, and autonomic dysfunction. The results indicate that fluctuating viral DNA load correlates with ME/CFS symptoms: this is in accordance with the hypothesis that pathogenesis is related to herpesvirus reactivation state, and this should be formally tested. Herpesvirus reactivation might be a cause or consequence of dysregulated immune function seen in ME/CFS. The sampling strategy and molecular tools developed here permit such large-scale epidemiological investigations.


Effect of Weight Loss via Severe vs Moderate Energy Restriction on Lean Mass and Body Composition Among Postmenopausal Women With Obesity: The TEMPO Diet Randomized Clinical Trial.
Severely energy-restricted diets are the most effective dietary obesity treatment. However, there are concerns regarding potential adverse effects on body composition. To compare the long-term effects of weight loss via severe vs moderate energy restriction on lean mass and other aspects of body composition. The Type of Energy Manipulation for Promoting Optimum Metabolic Health and Body Composition in Obesity (TEMPO) Diet Trial was a 12-month, single-center, randomized clinical trial. A total of 101 postmenopausal women, aged 45 to 65 years with body mass index (calculated as weight in kilograms divided by height in meters squared) from 30 to 40, who were at least 5 years after menopause, had fewer than 3 hours of structured physical activity per week, and lived in the Sydney metropolitan area of New South Wales, Australia, were recruited between March 2013 and July 2016. Data analysis was conducted between October 2018 and August 2019. Participants were randomized to either 12 months of moderate (25%-35%) energy restriction with a food-based diet (moderate intervention) or 4 months of severe (65%-75%) energy restriction with a total meal replacement diet followed by moderate energy restriction for an additional 8 months (severe intervention). Both interventions had a prescribed protein intake of 1.0 g/kg of actual body weight per day, and physical activity was encouraged but not supervised. The primary outcome was whole-body lean mass at 12 months after commencement of intervention. Secondary outcomes were body weight, thigh muscle area and muscle function (strength), bone mineral density, and fat mass and distribution, measured at 0, 4, 6, and 12 months. A total of 101 postmenopausal women were recruited (mean [SD] age, 58.0 [4.2] years; mean [SD] weight, 90.8 [9.1] kg; mean [SD] body mass index, 34.4 [2.5]). Compared with the moderate group at 12 months, the severe group lost more weight (effect size, -6.6 kg; 95% CI, -8.2 to -5.1 kg), lost more whole-body lean mass (effect size, -1.2 kg; 95% CI, -2.0 to -0.4 kg), and lost more thigh muscle area (effect size, -4.2 cm2; 95% CI, -6.5 to -1.9 cm2). However, decreases in whole-body lean mass and thigh muscle area were proportional to total weight loss, and there was no difference in muscle (handgrip) strength between groups. Total hip bone mineral density (effect size, -0.017 g/cm2; 95% CI, -0.029 to -0.005 g/cm2), whole-body fat mass (effect size, -5.5 kg; 95% CI, -7.1 to -3.9 kg), abdominal subcutaneous adipose tissue (effect size, -1890 cm3; 95% CI, -2560 to -1219 cm3), and visceral adipose tissue (effect size, -1389 cm3; 95% CI, -1748 to -1030 cm3) loss were also greater for the severe group than for the moderate group at 12 months. Severe energy restriction had no greater adverse effect on relative whole-body lean mass or handgrip strength compared with moderate energy restriction and was associated with 2-fold greater weight and fat loss over 12 months. However, there was significantly greater loss of total hip bone mineral density with severe vs moderate energy restriction. Therefore, caution is necessary when implementing severe energy restriction in postmenopausal women, particularly those with osteopenia or osteoporosis. anzctr.org.au Identifier: 12612000651886.


Bit1 Silencing Enhances the Proliferation, Migration, and Invasion of Glioma Cells Through Activation of the IL-6/STAT3 Pathway.
Several studies have indicated that the anoikis effector Bcl-2 inhibitor of transcription 1 (Bit1) can promote or inhibit tumor progression depending on the nature of the malignancy. However, its regulatory effects on gliomas are unknown. This study aimed at assessing Bit1 expression in glioma tissues and cells, its subsequent effects on glioma cell apoptosis, proliferation, invasion, and migration, and the underlying molecular mechanisms. The findings showed that lower Bit1 expressions in glioma tissues as well as a negative correlation between Bit1 expression and glioma grade. Additional findings also revealed that Bit1 silencing significantly inhibited anoikis and enhanced glioma cell proliferation, invasion, and migration. Further analysis showed that the decrease in Bit1 expressions led to malignancy proliferation and anoikis resistance through activation of the IL-6/STAT3 signaling pathway. Our data suggested that Bit1 may play an anti-oncogenic role in glioma cells and that a decrease in its expressions might induce glioma cell proliferation, migration, and invasion through the IL-6/STAT3 signaling pathway.


Characterisation of P2Y(12) receptor responsiveness to cysteinyl leukotrienes.
Leukotriene E4 (LTE4), the most stable of the cysteinyl leukotrienes (cysLTs), binds poorly to classical type 1 and 2 cysLT receptors although in asthmatic individuals it may potently induce bronchial constriction, airway hyperresponsiveness and inflammatory cell influx to the lung. A recent study has suggested that the purinergic receptor P2Y12 is required for LTE4 mediated pulmonary inflammation in a mouse model of asthma and signals in response to cysLTs. The aim of the study was to characterise the responsiveness of human P2Y12 to cysteinyl leukotrienes. Models of human CysLT1, CysLT2 and P2Y12 overexpressed in HEK293, CHO cells and human platelets were used and responsiveness to different agonists was measured using intracellular calcium, cAMP and β-arrestin recruitment assays. CysLTs induced concentration dependent calcium mobilisation in cells overexpressing CysLT1 and CysLT2 but failed to induce any calcium response in cells expressing P2Y12 or P2Y12+ Gα16. In contrast, selective P2Y12 agonists ADP and 2-MeS-ADP induced specific calcium flux in cells expressing P2Y12+ Gα16. Similarly, specific response to 2-MeS-ADP, but not to cysLTs was also observed in cells expressing P2Y12 when intracellular cAMP and β-arrestin signalling were analysed. Platelets were used as a model of human primary cells expressing P2Y12 to analyse potential signalling and cell activation through P2Y12 receptor or receptor heterodimers but no specific LTE4 responses were observed. These results show that LTE4 as well as other cysLTs do not activate intracellular signalling acting through P2Y12 and suggest that another LTE4 specific receptor has yet to be identified.


Experimental study on the vascular thermal response to visible laser pulses.
Port-wine stains (PWSs) are congenital vascular malformations that progressively darken and thicken with age, and laser therapy is the most effective in clinical practice. Using dorsal skin chamber (DSC), this study evaluated thermal response of blood vessel to a 595-nm pulsed dye laser (PDL) with controlled energy doses and pulse durations. Totally, 32 vessels (30∼300 μm in diameter) are selected from the dorsal skin of the mouse to match those in port-wine stain. The experimental results showed that the thermal response of the blood vessels to laser irradiation can be recognized as coagulation, constriction with diameter decrease, disappearance (complete constriction), hemorrhage, and collagen damage in the order of increasing laser radiant exposure. Blood vessels with small diameter would response poorly and survive from the laser heating because their thermal relaxation time is much shorter than the pulse duration. The optimalradiant exposure is from 10 to 12 J/cm(2) under 6 ms pulse duration without considering the epidermal light absorption. Numerical simulations were also conducted using a 1,000-μm deep Sprague-Dawley (SD) mouse skinfold. The light transportation and heat diffusion in dorsal skin were simulated with the Monte Carlo method and heat transfer equation, while the blood vessel photocoagulation was evaluated by Arrhenius-type kinetic integral. Both experimental observation and numerical simulation supported that hemorrhage is the dominant thermal response, which occurs due to preferential heating of the superior parts of large blood vessels. In clinical practice for 595 nm PDL, the consequent purpura caused by hemorrhage can be used as a treatment end point.


Congenital conduction defects in children born to asymptomatic mothers with anti-SSA/SSB antibodies: report of two cases.
It is very difficult to identify pregnant asymptomatic mothers carrying anti-SSA/SSB antibodies. We report two cases of neonatal lupus erythematosus, born to asymptomatic mothers with anti-SSA/Ro antibody, who developed isolated complete congenital cardiac heart block and transient second degree conduction defect associated with cardiac abnormalities, respectively. The first died suddenly of acute myocarditis at the age of 20 months, while the second underwent surgery at the age of 10 years for a ventricular septal defect, after two episodes of second degree atrioventricular block in infancy. We believe that in both cases the diagnosis could have been made in utero after correct heart beat analysis. We propose careful monitoring of fetal heart beat in all pregnant mothers. The occurrence of heart beat modification should prompt clinicians to test the mother for antibody positivity. This approach may permit early diagnosis and in utero treatment in order to spare the child from cardiac conduction defects. We provide the evidence of these cases and propose a flow chart for all physicians dealing with pregnancy.


Analysis of the Contribution of 6-mer Seed Toxicity to HIV-1-Induced Cytopathicity.
HIV-1 (HIV) infects CD4+ T cells, the gradual depletion of which can lead to AIDS in the absence of antiretroviral therapy (ART). Some cells, however, survive HIV infection and persist as part of the latently infected reservoir that causes recurrent viremia after ART cessation. Improved understanding of the mechanisms of HIV-mediated cell death could lead to a way to clear the latent reservoir. Death induced by survival gene elimination (DISE), an RNA interference (RNAi)-based mechanism, kills cells through short RNAs (sRNAs) with toxic 6-mer seeds (positions 2 to 7 of sRNA). These toxic seeds target the 3' untranslated region (UTR) of mRNAs, decreasing the expression of hundreds of genes critical for cell survival. In most cells under normal conditions, highly expressed cell-encoded nontoxic microRNAs (miRNAs) block access of toxic sRNAs to the RNA-induced silencing complex (RISC) that mediates RNAi, promoting cell survival. HIV has been shown to inhibit the biogenesis of host miRNAs in multiple ways. We now report that HIV infection of cells deficient in miRNA expression or function results in enhanced RISC loading of an HIV-encoded miRNA HIV-miR-TAR-3p, which can kill cells by DISE through a noncanonical (positions 3 to 8) 6-mer seed. In addition, cellular RISC-bound sRNAs shift to lower seed viability. This also occurs after latent HIV provirus reactivation in J-Lat cells, suggesting independence of permissiveness of cells to viral infection. More precise targeting of the balance between protective and cytotoxic sRNAs could provide new avenues to explore novel cell death mechanisms that could be used to kill latent HIV. IMPORTANCE Several mechanisms by which initial HIV infection is cytotoxic to infected cells have been reported and involve various forms of cell death. Characterizing the mechanisms underlying the long-term survival of certain T cells that become persistent provirus reservoirs is critical to developing a cure. We recently discovered death induced by survival gene elimination (DISE), an RNAi-based mechanism of cell death whereby toxic short RNAs (sRNAs) containing 6-mer seed sequences (exerting 6-mer seed toxicity) targeting essential survival genes are loaded into RNA-induced silencing complex (RISC) complexes, resulting in inescapable cell death. We now report that HIV infection in cells with low miRNA expression causes a shift of mostly cellular RISC-bound sRNAs to more toxic seeds. This could prime cells to DISE and is further enhanced by the viral microRNA (miRNA) HIV-miR-TAR-3p, which carries a toxic noncanonical 6-mer seed. Our data provide multiple new avenues to explore novel cell death mechanisms that could be used to kill latent HIV.


Nephroprotective effect of diosmin against sodium arsenite-induced renal toxicity is mediated via attenuation of oxidative stress and inflammation in mice.
Arsenic compounds, which are used in different industries like pesticide manufacturing, cause severe toxic effects in almost all organs, including the kidneys. Since the primary route of exposure to arsenic is through drinking water, and millions of people worldwide are exposed to unsafe levels of arsenic that can pose a threat to their health, this research was performed to investigate the nephroprotective effects of Diosmin (Dios), a flavonoid found in citrus fruits, against nephrotoxicity induced by sodium arsenite (SA). To induce nephrotoxicity, SA (10 mg/kg, oral gavage) was administered to mice for 30 days. Dios (25, 50, and 100 mg/kg, oral gavage) was given to mice for 30 days prior to SA administration. After the study was completed, animals were euthanized and blood and kidney samples were taken for biochemical and histopathological assessments. Results showed that SA-treated mice significantly increased the blood urea nitrogen and creatinine levels in the serum. This increase was associated with significant kidney tissue damage in SA-treated mice, which was confirmed by histopathological studies. Furthermore, SA enhanced the amounts of renal thiobarbituric acid reactive substances and decreased total thiol reserves, as well as the activity of antioxidant enzymes such as catalase, superoxide dismutase, and glutathione peroxidase. Also, in the SA-exposed group, an increase in the levels of kidney inflammatory biomarkers, including nitric oxide and tumor necrosis factor-alpha was observed. The western blot analysis indicated an elevation in the protein expression of kidney injury molecule-1 and nuclear factor-kappa B in SA-treated mice. However, pretreatment with Dios ameliorated the SA-related renal damage in mice. Our findings suggest that Dios can protect the kidneys against the nephrotoxic effects of SA by its antioxidant and anti-inflammatory characteristics.


Validity Evidence of Brazilian Portuguese Version of the Professional Fulfillment Index.
The questionnaire "Professional Fulfillment Index" (PFI) was validated to assess emotional exhaustion, interpersonal disengagement, and professional achievement among physicians. This study presents the process of cross-cultural adaptation of the PFI to the Brazilian context. Analytical cross-sectional study carried out between July and October 2020. Settings: For conceptual equivalence, a panel of experts was constituted who conducted analyzes on the translation and back-translation processes from English to Brazilian Portuguese. For the operational equivalence, 432 physicians in the field of Occupational Medicine were invited. They answered the PFI in an online platform. Analysis: Both exploratory and confirmatory factor analysis were performed to seek evidence of the hypothesized structure of PFI item responses. Reliability was assessed using three indicators. The Brazilian version can be considered similar to the original. Most of participants were female (57.6%) and mean age 49.6 years-SD 14.0. The mean time since graduation was 18.5 years-SD 9.67. Most of them had a full-time job (88.0%) and were trained through post-graduation courses 83.8%. The most frequent job task was to perform worker´s clinical evaluations (84.9%). Exploratory and confirmatory analyzes showed the adequacy of the items in measuring the instrument's construct, with stability for use in other populations and samples. The PFI was developed to evaluate positive and negative aspects of physicians' role and performance. The process described in this article was the first cross-cultural adaptation of the questionnaire. The PFI can help in mapping potential risk situations for negative impacts on physicians' occupational well-being and performance at work, in order to propose interventions that reduce exhaustion and expand professional fulfillment.


Lake browning counteracts cyanobacteria responses to nutrients: Evidence from phytoplankton dynamics in large enclosure experiments and comprehensive observational data.
Lakes worldwide are affected by multiple stressors, including climate change. This includes massive loading of both nutrients and humic substances to lakes during extreme weather events, which also may disrupt thermal stratification. Since multi-stressor effects vary widely in space and time, their combined ecological impacts remain difficult to predict. Therefore, we combined two consecutive large enclosure experiments with a comprehensive time-series and a broad-scale field survey to unravel the combined effects of storm-induced lake browning, nutrient enrichment and deep mixing on phytoplankton communities, focusing particularly on potentially toxic cyanobacterial blooms. The experimental results revealed that browning counteracted the stimulating effect of nutrients on phytoplankton and caused a shift from phototrophic cyanobacteria and chlorophytes to mixotrophic cryptophytes. Light limitation by browning was identified as the likely mechanism underlying this response. Deep-mixing increased microcystin concentrations in clear nutrient-enriched enclosures, caused by upwelling of a metalimnetic Planktothrix rubescens population. Monitoring data from a 25-year time-series of a eutrophic lake and from 588 northern European lakes corroborate the experimental results: Browning suppresses cyanobacteria in terms of both biovolume and proportion of the total phytoplankton biovolume. Both the experimental and observational results indicated a lower total phosphorus threshold for cyanobacterial bloom development in clearwater lakes (10-20 μg P L-1 ) than in humic lakes (20-30 μg P L-1 ). This finding provides management guidance for lakes receiving more nutrients and humic substances due to more frequent extreme weather events.


The composition and friction-reducing properties of leaf layers.
Every autumn rail networks across the world suffer delays, accidents and schedule changes due to low friction problems caused by leaves landing on the rails. These leaves form a layer that can reduce the friction between the wheel and the rail to a similar level as that between ice and an ice-skate ( μ = 0.01 - 0.05 ). Previous works have generated several hypotheses for the chemical reactions and low friction mechanism associated with these layers. In this work, the reaction between an aqueous extract of sycamore leaves and metallic iron is investigated. This reaction has been shown to produce a black precipitate, which matches field observations of leaf layers, while friction tests with these extracts produce characteristic ultra low friction. The reaction is investigated through FTIR, XPS, CHNS and ICP-MS analysis as well as wet chemical testing. The impact of the reaction on friction is investigated through three rounds of tribological testing. The results indicate that the black precipitate produced is iron tannate, formed by complexation of tannins with dissolved iron ions. Friction testing showed that eliminating tannins from the leaf extract resulted in a significant increase in the friction coefficient compared with the control.


Overall Survival of Black and White Men With Metastatic Castration-Resistant Prostate Cancer Treated With Docetaxel.
Several studies have reported that among patients with localized prostate cancer, black men have a shorter overall survival (OS) time than white men, but few data exist for men with advanced prostate cancer. The primary goal of this analysis was to compare the OS in black and white men with metastatic castration-resistant prostate cancer (mCRPC) who were treated in phase III clinical trials with docetaxel plus prednisone (DP) or a DP-containing regimen. Individual participant data from 8,820 men with mCRPC randomly assigned in nine phase III trials to DP or a DP-containing regimen were combined. Race was based on self-report. The primary end point was OS. The Cox proportional hazards regression model was used to assess the prognostic importance of race (black v white) adjusted for established risk factors common across the trials (age, prostate-specific antigen, performance status, alkaline phosphatase, hemoglobin, and sites of metastases). Of 8,820 men, 7,528 (85%) were white, 500 (6%) were black, 424 (5%) were Asian, and 368 (4%) were of unknown race. Black men were younger and had worse performance status, higher testosterone and prostate-specific antigen, and lower hemoglobin than white men. Despite these differences, the median OS was 21.0 months (95% CI, 19.4 to 22.5 months) versus 21.2 months (95% CI, 20.8 to 21.7 months) in black and white men, respectively. The pooled multivariable hazard ratio of 0.81 (95% CI, 0.72 to 0.91) demonstrates that overall, black men have a statistically significant decreased risk of death compared with white men ( P < .001). When adjusted for known prognostic factors, we observed a statistically significant increased OS in black versus white men with mCRPC who were enrolled in these clinical trials. The mechanism for these differences is not known.


Afferent C-fibres release substance P and glutamate.
Capsaicin is currently used as a specific pharmacological tool for investigation of functions of primary afferent C-fibres. Their peripheral terminals play an important role in "neurogenic inflammation," mediated by released substance P and calcitonin gene related peptide, whereas in the mediation of central functions, activation of the N-methyl-D-aspartate (NMDA) receptors has recently been demonstrated. A method for continuous monitoring of glutamate concentration was used to study mechanisms of capsaicin-induced glutamate release from rat spinal cord slices. Both capsaicin and substance P released glutamate from spinal dorsal horns in a concentration-dependent manner (EC50 = 0.53 +/- 0.07 and 0.37 +/- 0.06 microM, respectively). The NMDA antagonist MK-801 (10 microM) had no effect on evoked glutamate release, whereas the tachykinin (NK-1) antagonist CP-99994 (10 microM) reduced responses to both stimuli (p < 0.001). In capsaicin-desensitized rats, evoked glutamate release from dorsal horns was significantly decreased yet not completely abolished. Although the evoked glutamate release from ventral horns was markedly smaller than that from dorsal horns, the normalized responses to capsaicin and to substance P were similar. This might be explained by smaller amounts of mobilizable glutamate in ventral horns. Our findings confirmed the ability of capsaicin to release glutamate mainly from the afferent C-fibres in the spinal cord. The observed effect of exogenous substance P and inhibitory action of the NK-1 antagonist indicate facilitation of capsaicin-induced glutamate release by coreleased substance P.


In vitro effects of Apixaban on 5 different cancer cell lines.
Cancer is associated with hypercoagulability. However, several data suggest that anticoagulant drugs may have an effect on tumor development and progression mediated by both coagulation dependent processes and non-coagulation dependent processes. Therefore, we investigated the in vitro effects of Apixaban on cell proliferation, mortality, cell migration, gene expression and matrix metalloproteinase in 5 different cancer cell lines. The following cancer cell lines, and 2 normal fibroblast cultures (lung and dermal fibroblasts), were studied: OVCAR3 (ovarian cancer), MDA MB 231 (breast cancer), CaCO-2 (colon cancer), LNCaP (prostate cancer) and U937 (histiocytic lymphoma). Proliferation and cell mortality were assessed in control cells and Apixaban treated cultures (dose from 0.1 to 5 μg/ml, 0 to 96-h). Necrosis/Apoptosis (fluorescence microscopy), cell migration (24-h after scratch test), matrix metalloproteinase (MMP) activity and mRNA expression (RT PCR) of p16, p21, p53 and HAS were also assessed. High-dose (5 μg/ml) Apixaban incubation was associated with a significantly reduced proliferation in 3 cancer cell lines (OVCAR3, CaCO-2 and LNCaP) and with increased cancer cell mortality in all, except LNCaP, cancer lines. Apoptosis seems to account for the increased mortality. The migration capacity seems to be impaired after high-dose Apixaban incubation in OVCAR3 and CaCO-2 cells. Data on mRNA expression suggest a consistent increase in tumor suppression gene p16 in all cell lines. Our data suggest that high-dose Apixaban may be able to interfere with cancer cell in vitro, reducing proliferation and increasing cancer cell mortality through apoptosis in several cancer cell lines.


Next-generation sequencing for cytomegalovirus genotypic antiviral resistance testing.
Sanger-based sequencing has long served as the gold standard for detecting cytomegalovirus (CMV) resistance mutations. However, next-generation sequencing (NGS) offers a highly multiplexed and sensitive approach. Clinical implementation of NGS-antiviral resistance testing requires thorough validation. In this issue of the Journal of Clinical Microbiology, M. A. Mallory, W. C. Hymas, K. E. Simmon, M. T. Pyne, et al. (J Clin Microbiol 61:e00829-23, 2023, https://doi.org/10.1128/jcm.00829-23) detail a validation of a targeted NGS-based assay for multiple genomic regions that confer resistance to CMV antivirals and share their bioinformatics analysis and reporting pipeline. This validation can serve as guidance for laboratories wishing to develop similar methodologies.


Intraductal papillary mucinous neoplasms of the pancreas (IPMNs): epidemiology, diagnosis and future aspects.
Intraductal papillary mucinous neoplasms of the pancreas (IPMNs) are potentially malignant intraductal epithelial neoplasms which consist of columnar, mucin-containing cells and arise from the epithelium of the main pancreatic duct or its branches. IPMNs as well as pancreatic intraepithelial neoplasias (PanINs) and mucinous cystic neoplasms represent noninvasive precursors of invasive ductal adenocarcinoma of the pancreas. The diagnosis of IPMNs includes radiographic (CT scanning, MRI, MRCP) and endoscopic evaluation (ERCP, EUS), PET, as well as serum tumor markers and molecular markers. The Sendai Consensus Guidelines help guide surgical resection for patients with IPMN. The follow-up of these patients, as well as of those who do not undergo surgical resection, is of great importance, since patients with IPMN appear to be at risk for other malignancies. Herein, the authors summarize the data presented at the 2013 ASCO Gastrointestinal Cancers Symposium regarding incidence and clinicopathological characteristics of IPMN (Abstracts #324, #187 and #179).


Dibucaine stimulation of p-aminohippurate accumulation in rat kidney cortical slices.
We studied the effects of dibucaine on p-aminohippurate (PAH) transport using rat kidney cortical slices. Addition of dibucaine in concentrations from 0.01 to 0.1 mM stimulated PAH accumulation in a concentration-dependent way; at higher concentrations, accumulation was inhibited. Kinetic studies showed that dibucaine increased the apparent maximal activity of PAH transport, whereas the apparent Km remained essentially constant. The efflux of PAH from the slices was significantly depressed by the drug. Dibucaine had no significant effect on the sodium gradient, medium to intracellular fluid. The drug had biphasic effects on hypotonic hemolysis; at lower concentrations (up to 0.2 mM) it protected erythrocytes, and at much higher concentrations, it has a lytic effect. The pattern of the membrane-stabilizing action of dibucaine was similar to that of the stimulation of PAH accumulation. These results indicate that dibucaine markedly stimulated accumulation of PAH. Stimulation by this drug of PAH transport cannot be explained by the increase in the sodium gradient; it may be due at least in part to membrane stabilization.


Dietary L-arginine and cutaneous wound healing.
Skin wound healing has been the subject of extensive studies and various drugs have been used in an attempt to improve wound healing. There are conflicting data regarding the effects of L-arginine, the substrate of nitric oxide, on wound healing. We examined the 1-week rate of cutaneous wound healing and collagen deposition in three groups of rats who received a (1) L-arginine (2% in drinking water)-supplemented diet from three days before until the seventh day following injury (Group 1), (2) L-arginine-supplemented diet for three days before injury (Group 2), and (3) a standard diet without L-arginine supplementation (Group 3). The wound length and width were measured each day and then the open wound area and cumulative percentage of open wound area reduction were calculated. Wound biopsy samples were examined with Trichrome-Masson stain in a subgroup of animals. Results showed that Group 1 rats had a significantly lower cumulative percentage of open wound area reduction on day 7 compared to other two groups (Mann-Whitney U test, P < 0.05). Relatively higher degrees of wound collagen deposit (day 7) were noted in groups 2 and 3. It may be concluded that L-arginine (2% in water) administered three days before until the seventh day following skin wound induction may diminish the rate of skin wound healing and collagen deposition.


Motivations to nurse: an exploration of what motivates students in Pacific Island countries to enter nursing.
The aim of this study was to explore the motivations of student nurses enrolled in nursing courses across a variety of Pacific Island countries. The image of nursing, the desire to help others, family and friends in the profession, personal experience, security, travel opportunities and flexibility have all been identified as motivators for people to enter nursing. To date, what motivates students in Pacific Island countries to enrol in a nursing course has not been investigated. An exploratory qualitative approach using focus group interviews with 152 nursing students was undertaken. Data were analysed using thematic content analysis, revealing four themes: (i) helping others; (ii) 'making a difference for my people'; (iii) following in the footsteps of others; and (iv) financial and professional gain. In a time of health and nursing workforce shortages, developing a deeper understanding of what drives people can be used to improve recruitment strategies in the future.


[Immunomodulation of Uncaria tomentosa over dendritic cells, il-12 and profile TH1/TH2/TH17 in breast cancer].
Objetives. This study aimed to research the in vitro immunomodulatory effects of an Uncaria tomentosa hydroalcoholic extract standardized (5.03%, pentacyclic oxindole alkaloids) (UT-POA) on the immunophenotype of dendritic cells (DC) subsets, Th1, Th2, Th17 and IL-12 cytokines from patients with stage II breast cancer (BCII) and healthy women (H). Blood of 11 H and 7 BCII was obtained, PBMC were isolated and cultured for 2h with/without various concentrations of UT-POA and stimulated or not with LPS for 24h. PBMC were labeled with specific antibodies for DC and in the supernatant we measured Th1/Th2/Th17 cytokines, both by flow cytometry. Furthermore IL-12 was measured by ELISA. UT-POA did not alter DC or accessory molecules expression in BCII. However, H exhibited a decrease in the percentage of mDC (myeloid DC) and an increase in HLA-DR and CD86 expression at 1000 μg/mL. IL-12 secretion was modified only in the H group, increasing p70 subunit and decreasing p40 subunit. UT-POA increased Th1 (IFN-γ and IL-2), Th2 (IL-4) and Th17 (IL-17) secretion in both groups. UT-POA increased the production of cytokines related with anti-tumoral response at concentrations of 500-1000 μg/mL. This positive effect should be evaluated not only systemically but also in the tumor microenvironment in further studies. UT-POA may be a useful phytochemical in chemoprevention and in the alternative use in cancer therapies.


Comparative studies of mitral valves in rheumatic heart disease.
Mitral valve tissues from 195 patients with rheumatic heart disease removed at the time of cardiac surgery were studied by conventional histologic techniques. One hundred seven patients from a five-year period at All-India Institute for Medical Sciences in New Delhi were studied in conjunction with 88 patients undergoing cardiac surgery in Albuquerque during a similar period. In Indian patients a correlation was noted between degree of moderate or severe valvular calcification and extent of adjacent valvular mononuclear cellular infiltration often consisting of striking lymphocytic-plasma cell collections. Similar distinct correlations were not observed among the mitral valves from the Albuquerque patients. It is possible that mitral valve mononuclear cell infiltrates among Indian patients may reflect a chronic immune reaction influencing the natural course of rheumatic heart disease in this latter population.


BASELINE CORTICOSTERONE, HEMATOLOGY, AND BIOCHEMISTRY RESULTS AND CORRELATIONS TO REPRODUCTIVE SUCCESS IN NESTING LOGGERHEAD SEA TURTLES ( CARETTA CARETTA).
Characterizing the health status and reproductive success of wild populations of sea turtles can be difficult; however, obtaining data to do this can provide important insight into the stability and long-term success of a population. This study examined the use of baseline corticosterone to assess reproductive success of a population of nesting loggerhead sea turtles ( Caretta caretta) on Jekyll Island, Georgia and investigated hematological and biochemical trends in this population. A total of 37 nesting loggerhead sea turtles was sampled for this study. Eleven (29.7%) turtles were sampled in 2013 and 26 (70.3%) were sampled in 2014. A majority of the turtles sampled successfully nested (29/37, 78.4%; false crawls: 8/37, 21.6%). There was no significant correlation between baseline corticosterone concentrations and reproductive success (hatch success or emergence success) in this population. There was a significant positive correlation ( r = 0.461, P = 0.02) between corticosterone concentrations and absolute monocyte counts. There was a significant negative correlation between monocyte count and hatch success ( r = -0.464, P = 0.05) and a positive correlation between phosphorus and hatch success ( r = 0.405, P = 0.05). Calcium concentrations were significantly different ( P = 0.01) between animals that false crawled and those that nested, with nesting turtles having lower calcium concentrations than those that false crawled. Turtles that false crawled were significantly ( P = 0.008) more likely to have elevated potassium concentrations than turtles that nested. This study provides the first attempt at characterizing baseline corticosterone, hematology, and biochemistry data and correlations with reproductive success in nesting loggerhead sea turtles. Overall, loggerhead sea turtles capable of nesting were found to be in good health and have good reproductive success while maintaining low levels of corticosterone during reproductive activities.


Aging of the auditory system.
Presbycusis or age-related hearing loss (ARHL) affects most elderly people. It is characterized by reduced hearing thresholds and speech understanding with the well-known negative consequences for communication and quality of social life. The hearing loss is connected to age-related histologic changes, as described and classified by Schuknecht. Aging itself is a multifactorial, genetically driven process that is influenced by oxidative stress that gradually leads to reduced endocochlear potential and cell loss of key players in sound transmission and supporting structures. Oxidative stress is caused by damaging factors like noise, infection, and other systemic factors. All reparative mechanisms in acute and chronic cochlear damage attempt to reduce oxidative stress and to balance inner-ear homeostasis. Accurate clinical assessment of ARHL starts with the differentiation between peripheral and central components. Treatment of the peripheral hearing loss often involves hearing aids, whereas auditory and psychologic training seems to be important in central auditory disturbance.