The first clinical and epidemiological programme on renal disease in Bolivia: a model for prevention and early diagnosis of renal diseases in the developing countries.
The prevalence and incidence of renal diseases in developing countries are not known. This lack of knowledge is an obstacle to the adoption of preventive measures which may be of great value in a social and economic environment where treatment options for end-stage renal failure are simply not available to the vast majority of the population. Urinalysis, a simple and inexpensive test, remains a cornerstone in the evaluation of the kidney and may also be easily employed in mass screening for renal abnormalities in a developing country. An educational campaign on renal diseases was conducted in three selected areas of Bolivia. Urine samples were collected and sent to one of 21 participating clinical centers. Fresh urine specimens were screened using a dipstick for chemical analysis and by microscopic urinalysis after centrifugation. In those patients in whom urinary abnormalities were found, further investigations were carried out in order to define the diagnosis; these patients were enrolled in a 3-year follow-up program. Apparently healthy subjects (n = 14,082) were referred to the First Clinical and Epidemiological Program of Renal Diseases from rural and metropolitan areas in Bolivia. Urinary abnormalities were detected in 4261 subjects at first screening. The most common form of urinary abnormality was hematuria, which was found in 2010 (47% of positively screened subjects). Other renal abnormalities were leukocyturia (41%) and proteinuria (11%). Confirmatory tests and further clinical studies were then carried out in 1019 people. On a second screening 35% of the subjects had no urinary abnormalities; in the remaining people the following diagnosis were made: asymptomatic urinary-tract infection (48.4%), isolated benign hematuria (43.9%), chronic renal failure (1.6%), renal tuberculosis (1.6%). Other diagnosis were: renal stones 1.3%, diabetic nephropathy 1% and polycystic kidney diseases 1.9%. This study helped define for the first time the frequency of asymptomatic renal diseases in Bolivia. It shows that it is possible to screen a large population of patients at relatively low cost, providing the framework for further action that may help in the prevention and timely diagnosis of renal diseases.


Zerumbone reduced the inflammatory response of acute lung injury in endotoxin-treated mice via Akt-NFκB pathway.
Zerumbone, a cyclic eleven-membered sesquiterpene, is the major component of the essential oil isolated from the wild ginger, Zingiber zerumbet. There are several beneficial pharmacological activities of zerumbone including anti-inflammatory, antioxidant, and anticancer activities. Acute lung injury (ALI) is an acute pulmonary inflammatory disorder with high morbidity and mortality rate. In present study, we aimed to investigate the protective effects and mechanisms of zerumbone on endotoxin, lipopolysaccharide (LPS)-induced ALI. Mice were pretreated with zerumbone at various concentrations for 30 min followed by intratracheal administration of LPS for 6 h. Pretreatment with zerumbone not only reduced leukocytes infiltration into the alveolar space but also inhibited lung edema in LPS-induced ALI. Decreased secretion of proinflammatory cytokines such as TNFα and IL-6 caused by LPS were reversed by zerumbone. LPS-induced expressions of proinflammatory mediators, iNOS and COX-2, were inhibited by zerumbone. In addition, NFκB activation and Akt phosphorylation were inhibited by zerumbone in LPS-induced ALI. All these results suggested that the protective mechanisms of zerumbone on endotoxin-induced ALI were via inhibition of Akt-NFκB activation.


Pain assessment in temporomandibular disorders.
Pain characteristics, symptoms, medication use, demographic and psychological variables were assessed in 85 patients diagnosed with temporomandibular (TM) disorders. Results showed that 46 myofascial pain dysfunction (MPD) patients responded differently from 39 temporomandibular joint (TMJ) patients for numerous measures of pain-related variables. MPD patients perceived more generalized facial pain of longer duration and were able to identify more specific symptoms associated with their pain. The only symptom variable rated higher by TMJ patients was the presence of bothersome joint noise. When asked to rate their pain at its worse, the unpleasantness of pain was significantly higher in MPD patients than reported by TMJ patients. MPD patients might be considered more psychologically distressed than TMJ patients, as shown by their higher ratings for anxiety and inability to endure the pain. Although both types of pain patients took the same categories of medications, MPD patients consumed a larger total number. Such findings suggest that the prognosis of the dysfunction for MPD patients would be less favorable than for TMJ patients.


Hematopoietic stem cells are uniquely selective in their migratory response to chemokines.
Although hematopoietic stem cell (HSC) migration into and out of sites of active hematopoiesis is poorly understood, it is a critical process that underlies modern clinical stem cell transplantation and may be important for normal hematopoietic homeostasis. Given the established roles of chemotactic cytokine (chemokine)-directed migration of other leukocyte subsets, the migration of murine HSC to a large panel of CC and CXC chemokines was investigated. HSC migrated only in response to stromal derived factor-1alpha, the ligand for the CXC chemokine receptor 4 (CXCR4). CXCR4 expression by HSC was confirmed by reverse transcription polymerase chain reaction analysis. Surprisingly, HSC also expressed mRNA for CCR3 and CCR9, although they failed to migrate to the ligands for these receptors. The sharply restricted chemotactic responsiveness of HSC is unique among leukocytes and may be necessary for the specific homing of circulating HSC to bone marrow, as well as for the maintenance of HSC in hematopoietic microenvironments.


Transcriptional regulation of type I diabetes by NF-kappa B.
Development of type I diabetes requires coordinated expression of myriad genes responsible for the initiation and progression of the disease. Expression of these genes are regulated by a small number of transcription factors including the Rel/NF-kappaB family. To determine the roles of the Rel/NF-kappaB family in type I diabetes, we studied multiple low-dose streptozotocin-induced diabetes in mice deficient in either c-Rel or NF-kappaB1. We found that mice deficient in each of these NF-kappaB subunits were resistant to streptozotocin-induced diabetes. However, the mechanisms of the disease resistance may differ in different cases. Deficiency in c-Rel selectively reduced Th1, but not Th2 responses, whereas NF-kappaB1 deficiency had little effect on T cell responses to anti-CD3 stimulation. Death of dendritic cells was accelerated in the absence of NF-kappaB1, whereas death of macrophages and granulocytes was affected primarily by c-Rel deficiency. Furthermore, Stat-1 expression was significantly reduced in macrophages deficient in NF-kappaB1, but not c-Rel. These results indicate that both c-Rel and NF-kappaB1are essential for the development of type I diabetes and that strategies targeting each of these subunits would be effective in preventing the disease.


Characterization of Transcription Termination-Associated RNAs: New Insights into their Biogenesis, Tailing, and Expression in Primary Tumors.
Next-generation sequencing has uncovered novel classes of small RNAs (sRNAs) in eukaryotes, in addition to the well-known miRNAs, siRNAs, and piRNAs. In particular, sRNA species arise from transcription start sites (TSSs) and the transcription termination sites (TTSs) of genes. However, a detailed characterization of these new classes of sRNAs is still lacking. Here, we present a comprehensive study of sRNAs derived from TTSs of expressed genes (TTSa-RNAs) in human cell lines and primary tissues. Taking advantage of sRNA-sequencing, we show that TTSa-RNAs are present in the nuclei of human cells, are loaded onto both AGO1 and AGO2, and their biogenesis does not require DICER and AGO2 endonucleolytic activity. TTSa-RNAs display a strong bias against a G residue in the first position at 5' end, a known feature of AGO-bound sRNAs, and a peculiar oligoA tail at 3' end. AGO-bound TTSa-RNAs derive from genes involved in cell cycle progression regulation and DNA integrity checkpoints. Finally, we provide evidence that TTSa-RNAs can be detected by sRNA-Seq in primary human tissue, and their expression increases in tumor samples as compared to nontumor tissues, suggesting that in the future, TTSa-RNAs might be explored as biomarker for diagnosis or prognosis of human malignancies.


MSX1 induces G0/G1 arrest and apoptosis by suppressing Notch signaling and is frequently methylated in cervical cancer.
The objectives of this study were to investigate the expression of MSX1 in cervical cells and tissues, the methylation status of the MSX1 promoter, the influence of overexpression of gene MSX1 on the proliferation, migration, and invasion of HeLa and SiHa cells, and finally the possible molecular mechanisms responsible for the suppressive effects of MSX1 upon cervical cancer cells. Semi-quantitative and quantitative reverse transcription-polymerase chain reactions were used to investigate the expression levels of MSX1, and methylation-specific polymerase chain reaction (MSP) was performed to investigate promoter methylation status in cervical cancer cell lines, primary cervical tissues, and normal cervical tissues. Clone formation, Cell Counting Kit-8 (CCK-8), cell wound scratch, and transwell assays were performed to verify whether MSX1 could inhibit the proliferation and migration of cervical cancer cells. Western blot was used to analyze the effect of MSX1 upon Notch1, Jagged1, c-Myc, cleaved PARP, cleaved caspse-3, and cyclin D1 (CCND1). MSX1 was frequently downregulated or silenced in 60.0% (3/5) of cervical cancer cell lines. The promoter methylation of MSX1 was detected in 42.0% (42/100) of primary tumor tissues, while no methylation was observed in normal cervical tissues. Pharmacological demethylation reduced MSX1 promoter methylation levels and restored the expression of MSX1. The overexpression of MSX1 in cervical cancer cells thus inhibited the proliferation and migration of cervical cancer cells. The overexpression of MSX1 in cervical cancer cells downregulated the expression levels of Notch1, Jagged1, and c-Myc but upregulated the expression levels of CCND1, cleaved PARP, and cleaved caspase-3. MSX1 appears to be a functional tumor suppressor that regulates tumorigenesis in cervical cancer by antagonizing Notch signaling.


Use of Cardioprotective Dexrazoxane Is Associated with Increased Myelotoxicity in Anthracycline-Treated Soft-Tissue Sarcoma Patients.
Dexrazoxane (DEX) is indicated as a cardioprotective agent for breast cancer patients receiving the anthracycline doxorubicin. Two meta-analyses in metastatic breast cancer reported an apparent increase in the severity of myelosuppression when DEX was used. So far, no data in soft-tissue sarcoma (STS) patients are available. We retrospectively analyzed hematological toxicity data from 133 consecutive STS patients who received a chemotherapy regimen containing an anthracycline and ifosfamide (AI) in the perioperative or metastatic settings between January 2006 and December 2017. Of these, 46 received off-label DEX concurrently with the AI treatment. The differences between incidence of any of the explored outcomes were assessed according to the Fisher exact test. Compared with the non-DEX group, DEX treatment was associated with significantly higher rates of grade 3/4 hematological toxicities: leukopenia (56.5 vs. 28.7%; p = 0.0014), neutropenia (69.6 vs. 24.1%; p = 0.0001), febrile neutropenia (52.2 vs. 20.7%; p = 0.0004), anemia (41.3 vs. 28.7%; p = 0.1758), and thrombocytopenia (54.3 vs. 32.1%; p = 0.0159). Similarly, in the DEX group dose reductions were more frequent compared to the non-DEX group (39.1 vs. 19.5%; p = 0.0221). Adding DEX to AI in STS patients leads to higher rates of bone marrow suppression in all blood components, as well as to more frequent events of febrile neutropenia and dose reductions.


Multiplexed nanoplasmonic biosensor for one-step simultaneous detection of Chlamydia trachomatis and Neisseria gonorrhoeae in urine.
Development of rapid and multiplexed diagnostic tools is a top priority to address the current epidemic problem of sexually transmitted diseases. Here we introduce a novel nanoplasmonic biosensor for simultaneous detection of the two most common bacterial infections: Chlamydia trachomatis and Neisseria gonorrhoeae. Our plasmonic microarray is composed of gold nanohole sensor arrays that exhibit the extraordinary optical transmission (EOT), providing highly sensitive analysis in a label-free configuration. The integration in a microfluidic system and the precise immobilization of specific antibodies on the individual sensor arrays allow for selective detection and quantification of the bacteria in real-time. We achieved outstanding sensitivities for direct immunoassay of urine samples, with a limit of detection of 300 colony forming units (CFU)/mL for C. trachomatis and 1500CFU/mL for N. gonorrhoeae. The multiplexing capability of our biosensor was demonstrated by analyzing different urine samples spiked with either C. trachomatis or N. gonorrhoeae, and also containing both bacteria. We could successfully detect, identify and quantify the levels of the two bacteria in a one-step assay, without the need for DNA extraction or amplification techniques. This work opens up new possibilities for the implementation of point-of-care biosensors that enable fast, simple and efficient diagnosis of sexually transmitted infections.


What derived variables should be monitored during mechanical ventilation?
Without a careful definition, it is very difficult to propose a list of essential derived variables that should be monitored during mechanical ventilation. The list of essentials will vary not only with disease type and severity but also with the expertise of the operator in interpreting the data, and willingness to incorporate it into his/her surveillance and treatment plan. It can be cogently argued that the only variables of crucial significance to the vast majority of patients are the primary ones--airway pressure, flow, tidal volume, and minute ventilation. My own view is that end-inspiratory (PD, Ps, and Pz), end-expiratory (total PEEP), and mean airway pressures must be checked at frequent intervals, especially in ARDS. Partitioning of the total pressure into its flow-driving and elastance-counterbalancing components is always wise, whether or not resistance and compliance or elastance are formally calculated. Incremental changes in the pressure-volume relationship should be monitored whenever adjustments in PEEP or VT are made. Ventilatory demand, strength, and power-reserve assessment are often instrumental in the care of the ventilator-dependent patient who presents as a weaning problem. The most valuable indicators of these include the VE, the maximum voluntary inspiratory pressure, and the frequency-to-tidal-volume ratio. Measurements of the work of breathing, P0.1, and Pes should be reserved for unusually difficult clinical questions. Finally, the variability of the Paw tracing yields valuable data regarding the synchrony of patient-ventilator interactions.


Profiling of normal human leg lymph proteins using the 2-D electrophoresis and SELDI-TOF mass spectrophotometry approach.
The parenchymatous cells are supplied by nutrients transported in fluid from blood across the capillary wall. This fluid, called tissue fluid (TF), contains proteins originating from plasma as well as those synthesized and secreted by tissue cells. The protein composition of TF remains largely unknown. The TF which has entered lymphatics is called lymph (L). Harvesting L and measuring its proteins concentrations and identifying them provide an insight into biochemical processes in the TF. Here we describe our initial evaluation of the normal human prenodal L protein profile of m.w. 2.5 to 12.5 kDa using the ProteinChip SELDI MS system and compare it with that of plasma (P) protein. This is the first study in the literature providing evidence for the presence of the so far non-identified proteins in L as well as proteins identified in L but absent from P and conversely present in P but not in L. Evident differences between paired L and P samples have been found, along with similarities. Thirteen proteins were detected in P and seven in L in the region of 2.5 to 12.5 kDa. Five identical proteins, although of different relative intensity, were found in L and P. The proteins specific for L but not P had 7070 and 8619 ion values. P proteins absent from L were of 3890, 3969, 4078, 6863, 7676, 7778, 7847 and 7937 ion values. In addition to detecting some so far unknown proteins in L, these preliminary findings throw a new light on our understanding of the mechanism of transcapillary transport of low m.w. proteins. They challenge the commonly accepted notion of unlimited free diffusion of peptides across the capillary membrane.


Are uterine and ovarian artery Doppler velocimetry values good pregnancy predictors in clomiphene citrate cycles?
We conducted this prospective study to evaluate the prognostic significance of uterine and ovarian artery Doppler velocimetry in clomiphene citrate (CC) cycles. A total of 80 patients with unexplained infertility were given 100 mg/day of CC from day 3 to day 7 of their cycles in this current prospective study. On cycle day 3, before administration of CC, each patient underwent Doppler transvaginal ultrasonography. The Doppler velocimetries of the right and left uterine and ovarian arteries were recorded and analyzed in association with demographic and clinical parameters. TheThere were 6 out of 80 patients who became pregnant, the overall pregnancy rate in this population was 7.5% for the current study. The cases were divided into two groups according to whether they became pregnant or not. Demographic characteristics showed no statistically significant differences between these groups (p>0.05). However, the duration of infertility did show statistically significant differences between the groups. Doppler velocimetry was not statistically significantly different between the two groups. Doppler velocimetry of the uterine and ovarian arteries is not a factor in the prognosis for pregnancy in CC cycles.


Management and outcome of consecutive pregnancies complicated by idiopathic intracranial hypertension.
The effects of consecutive pregnancies on the course of idiopathic intracranial hypertension (IIH) are unclear in view of the scarce published data. To evaluate the course and management of visual and pregnancy outcomes of consecutive pregnancies with IIH. The medical records of women with IIH in consecutive pregnancies were reviewed for neuro-ophthalmological findings, management, and visual and pregnancy outcomes. The study group comprised eight women with at least two consecutive pregnancies (mean age 27.3 +/- 5.3 years). The mean duration of IIH prior to the first pregnancy was 3.4 +/- 3.16 years. One woman with IIH pre-pregnancy symptoms and three women with clinical features of IIH during the second trimester of pregnancy (gestational week 21.7 +/- 4.04) were treated with acetazolamide (250 mg every 8 hours). Symptoms resolved, resulting in uncomplicated first deliveries for all four. The first deliveries of four other women were by cesarean section due to obstetric indications. Only one woman developed symptoms and signs of IIH during her second pregnancy and was thus treated with acetazolamide. Two women who completed three pregnancies had no IIH symptoms during their pregnancies. The course and outcome of those pregnancies were normal. IIH apparently does not worsen or even become symptomatic in consecutive pregnancies. The appropriate management of IIH in pregnant women is similar to management for non-pregnant women; neither the course nor the obstetric outcome of first and consecutive pregnancies is influenced by the presence of IIH.


Evaluation of SARS-CoV-2 in Breastmilk from 18 Infected Women.
To The Editor, Currently, the U.S. Centers for Disease Control and Prevention, American Academy of Pediatrics and the World Health Organization advise that women who are infected with SARS-CoV-2 may choose to breastfeed with appropriate protections to prevent transmission of the virus through respiratory droplets. However, the potential for exposure to SARS-CoV-2 through breastfeeding is currently unknown. To date, case reports on breastmilk samples from a total of 24 SARS-CoV-2-infected women have been published. Of those, viral RNA was detected in ten breastmilk samples from four women. In some but not all cases, environmental contamination as the source of the virus or retrograde flow from an infected infant could not be ruled out. We established a quantitative RT-PCR assay for SARS-CoV-2 in breastmilk with a limit of detection of 250 copies per mL and validated it by spiking breastmilk from uninfected women with known amounts of viral RNA. In addition, we established tissue culture methods to detect replication-competent SARS-CoV-2 in breastmilk. No viral RNA nor culturable virus was detected after Holder pasteurization of breastmilk samples that had been spiked with replication-competent SARS-CoV-2 (see Supplement). Between March 27 and May 6, 2020, we collected and analyzed 64 serial breastmilk samples from 18 SARS-CoV-2-infected women residing in the U.S. (see Supplement for clinical characteristics). Breastmilk samples were collected before and after women had a positive SARS-CoV-2 RT-PCR test and all but one woman had symptomatic disease (see Figure). One of the 64 breastmilk samples had detectable SARS-CoV-2 RNA by RT-PCR. The positive sample was collected on the day of symptom onset but one sample 2 days prior to symptom onset and two subsequent samples, collected 12 and 41 days later, tested negative for viral RNA. In addition, a subset of 26 breastmilk samples from nine women were tested for the presence of replication-competent virus using our established culture methods, and all were negative including the one sample that tested positive for viral RNA by RT-PCR. Although SARS-CoV-2 RNA was detected in one milk sample from one of eighteen infected women, the viral culture for that sample was negative. This suggests that SARS-CoV-2 RNA does not represent replication-competent virus and that breastmilk itself is likely not a source of infection for the infant. Furthermore, when control breastmilk samples spiked with replication-competent SARS-CoV-2 virus were treated by Holder pasteurization, a process commonly performed by donor milk banks, no replication-competent virus nor viral RNA was detectable. Further research to confirm these findings is needed, as well as an examination of convalescent milk for the presence of antibodies against SARS-CoV-2.


Intra-Articular Morphine versus Dexmedetomedine for Knee Arthroscopy under Local Anesthesia.
Knee arthroscopy has both diagnostic and therapeutic applications which can be performed under general, regional, or local anesthesia. Morphine is used as an additive to local anesthetics. Dexmedetomedine, the highly selective alpha-2 (α2)-adrenoceptor agonist with the sedative and analgesic effect can be used also to augment local anesthetic effect. Sixty patients submitted for elective knee arthroscopy whose age between 25 and 45 years, of either sex, the American society of anethesiologists physical status Classes I and II at a university hospital were enrolled in this study. Patients were classified into two groups. Morphine Group (M) (n = 30): Patients received 20 ml of 0.5% bupivacaine plus 5 ml of 0.2% lidocaine with epinephrine 1:200,000 plus 1 mg morphine. Dexmedetomedine Group (D) (n = 30): Patients received 20 ml of 0.5% bupivacaine plus 5 ml of 0.2% lidocaine with epinephrine 1:200,000 plus 1 μg/kg dexmedetomedine. Demographic data of patients showed no significant difference among the studied groups. Heart rate (HR) was significantly lower in (D) Group compared to that of (M) Group 5 min from the start of procedure to immediately postoperatively. Moreover, (D) Group showed a significant decrease in HR 10 min up to 35 min intraoperatively compared to the basal value. Furthermore, mean arterial blood pressure (MBP) was significantly lower in (D) Group compared to that of (M) Group 15 min from the start of surgery up to 1 h postoperatively. Furthermore, (D) Group showed a significant decrease in MBP 15 min intraoperative up to 2 h postoperatively compared to the basal value. While there was no significant difference in (visual analogue score [VAS], onset and total consumption of ketorolac, surgeon and patients' satisfaction, side effects in (D) Group compared to (M) Group, respectively. Addition of either morphine or dexmedetomidine to bupivacaine intraarticularly improved both intraoperative anesthesia and postoperative analgesia with minimal side effects or complications in knee arthroscopy, with superiority of dexmedetomidine compared to morphine on hemodynamic stability.


Occurrence of serological reactions for serogroup Sejroe (CTG and Prajtino) in female buffalo in the state of Pernambuco, Brazil.
The objective of this study was to evaluate the occurrence of anti-Leptospira spp. antibodies in female buffalo in the state of Pernambuco. A total of 123 female buffalo blood samples were collected from five properties distributed in the state of Pernambuco. The microscopic agglutination test was used to study anti-Leptospira spp. antibodies. The occurrence of anti-Leptospira spp. antibodies was 28.5% (35/123; CI 20.7-37.3%) and on different properties, the occurrence ranged from 28.6% to 80.0%, with 100% of the properties showing animals with positive results. The serovars of the serogroup Sejroe with a higher incidence were Hardjoprajtino (CTG strain, 49.1%) and Hardjo (Prajtino genotype, 43.2%), followed by serogroup Grippotyphosa with the Grippotyphosa serovar (3.9%), serogroup Pomona with the Pomona serovar (1.9%), and the Icterohaemorrhagiae serovar Copenhageni (1.9%). This was the first record of the occurrence of anti-Lepstospira spp. antibodies in female buffalo in the state of Pernambuco. Control measures are necessary to prevent health and economic losses, given that the agent involved affects animal reproduction, triggering drops in conception rates or even clinical cases of abortion.


Zebrafish Cx35: cloning and characterization of a gap junction gene highly expressed in the retina.
The vertebrate connexin gene family encodes protein subunits of gap junction channels, which provide a route for direct intercellular communication. Consequently, gap junctions play a vital role in many developmental and homeostatic processes. Aberrant functioning of gap junctions is implicated in many human diseases. Zebrafish are an ideal vertebrate model to study development of the visual system as they produce transparent embryos that develop rapidly, thereby facilitating morphological and behavioral testing. In this study, zebrafish connexin35 has been cloned from a P1 artificial chromosome (PAC) library. Sequence analysis shows a high degree of similarity to the Cx35/36 orthologous group, which are expressed primarily in nervous tissue, including the retina. The gene encodes a 304-amino acid protein with a predicted molecular weight of approximately 35 kDa. Injection of zebrafish Cx35 RNA into paired Xenopus oocytes elicited intercellular electrical coupling with weak voltage sensitivity. In development, Cx35 is first detectable by Northern analysis and RT-PCR, at 2 days post-fertilization (2 dpf), and in the adult it is expressed in the brain and retina. Immunohistochemical analysis revealed that the Cx35 protein is expressed in two sublaminae of the inner plexiform layer of the adult retina. A similar pattern was seen in the 4 and 5 dpf retina, but no labeling was detected in the retina of earlier embryos.


Autonomic dysregulation, cognition and fatigue in people with depression and in active and healthy controls: observational cohort study.
Autonomic nervous system (ANS) dysregulation might be relevant to the pathophysiology of fatigue and cognitive impairment in depression and perhaps should be considered when making prescribing decisions. To determine the relationship of self-reported ANS symptoms with fatigue, cognition and prescribed medication in people with a diagnosis of depression, in comparators without depression but with other mental health, neurodevelopmental or neurodegenerative disorders (active controls) and in healthy controls. Cross-sectional analysis of an opportunistic sample from England. Self-reported data were collected on demographics, diagnosis, medication, ANS symptoms (Composite Autonomic Symptom Scale-31, COMPASS-31) and fatigue (Visual Analogue Scale for Fatigue, VAS-F). A subsample completed cognitive tests (THINC-it), including the subjective Perceived Deficits Questionnaire five-item version (PDQ-5). Spearman's correlation and mediation models were used to explore the relationship between COMPASS-31, VAS-F and PDQ-5 scores. Data were obtained for 3345 participants, 22% with depression. The depression group had significantly (P < 0.01) more severe autonomic dysregulation as measured by COMPASS-31 scores (median 30) than active (median 23) and healthy controls (median 10). The depression group had significantly higher symptom severity (P < 0.01) than both control groups on the VAS-F and PDQ-5. Overall, there was a significantly positive correlation (P < 0.01) between COMPASS-31, VAS-F scores (Spearman's rho rs = 0.44) and PDQ-5 scores (rs = 0.56). COMPASS-31 scores mediated greater symptom severity on the VAS-F and PDQ-5 for those with depression. COMPASS-31 scores remained significantly different between the depression group and both control groups independently of medication. People with a diagnosis of depression report worse fatigue and cognition than active and healthy comparators; this appears to be mediated by ANS dysregulation.


Chronic venous disease in an ethnically diverse population: the San Diego Population Study.
In a 1994-1998 cross-sectional study of a multiethnic sample of 2,211 men and women in San Diego, California, the authors estimated prevalence of the major manifestations of chronic venous disease: spider veins, varicose veins, trophic changes, and edema by visual inspection; superficial and deep functional disease (reflux or obstruction) by duplex ultrasonography; and venous thrombotic events based on history. Venous disease increased with age, and, compared with Hispanics, African Americans, and Asians, non-Hispanic Whites had more disease. Spider veins, varicose veins, superficial functional disease, and superficial thrombotic events were more common in women than men (odds ratio (OR) = 5.4, OR = 2.2, OR = 1.9, and OR = 1.9, respectively; p < 0.05), but trophic changes and deep functional disease were less common in women (OR = 0.7 for both; p < 0.05). Visible (varicose veins or trophic changes) and functional (superficial or deep) disease were closely linked; 92.0% of legs were concordant and 8.0% discordant. For legs evidencing both trophic changes and deep functional disease, the age-adjusted prevalences of edema, superficial events, and deep events were 48.2%, 11.3%, and 24.6%, respectively, compared with 1.7%, 0.6%, and 1.3% for legs visibly and functionally normal. However, visible disease did not invariably predict functional disease, or vice versa, and venous thrombotic events occurred in the absence of either.


Differential effects of trichostatin A on mouse embryogenesis and development.
Trichostatin A (TSA), a histone deacetylase (HDAC) inhibitor, can significantly improve the reprogramming efficiency of somatic cells. However, whether TSA has a detrimental effect on other kinds of embryos is largely unknown because of the lack of integrated analysis of the TSA effect on natural fertilized embryos. To investigate the effect of TSA on mouse embryo development, we analyzed preimplantation and post-implantation development of in vivo, in vitro fertilized, and parthenogenetic embryos treated with TSA at different concentrations and durations. In vivo fertilized embryos appeared to be the most sensitive to TSA treatment among the three groups, and the blastocyst formation rate decreased sharply as TSA concentration and treatment time increased. TSA treatment also reduced the livebirth rate for in vivo fertilized embryos from 56.59 to 38.33% but did not significantly affect postnatal biological functions such as the pups' reproductive performance and their ability for spatial learning and memory. Further analysis indicated that the acetylation level of H3K9 and H4K5 was enhanced by TSA treatment at low concentrations, while DNA methylation appeared to be also disturbed by TSA treatment only at high concentration. Thus, our data indicates that TSA has different effects on preimplantation embryonic development depending on the nature of the embryo's reproductive origin, the TSA concentration and treatment time, whereas the effect of TSA at the indicated concentration on postnatal function was minor.


Identification of Suicide Attempt Risk Factors in a National US Survey Using Machine Learning.
Because more than one-third of people making nonfatal suicide attempts do not receive mental health treatment, it is essential to extend suicide attempt risk factors beyond high-risk clinical populations to the general adult population. To identify future suicide attempt risk factors in the general population using a data-driven machine learning approach including more than 2500 questions from a large, nationally representative survey of US adults. Data came from wave 1 (2001 to 2002) and wave 2 (2004 to 2005) of the National Epidemiologic Survey on Alcohol and Related Conditions (NESARC). NESARC is a face-to-face longitudinal survey conducted with a national representative sample of noninstitutionalized civilian population 18 years and older in the US. The cumulative response rate across both waves was 70.2% resulting in 34 653 wave 2 interviews. A balanced random forest was trained using cross-validation to develop a suicide attempt risk model. Out-of-fold model prediction was used to assess model performance, including the area under the receiver operator curve, sensitivity, and specificity. Survey design and nonresponse weights allowed estimates to be representative of the US civilian population based on the 2000 census. Analyses were performed between May 15, 2019, and June 10, 2020. Attempted suicide in the 3 years between wave 1 and wave 2 interviews. Of 34 653 participants, 20 089 were female (weighted proportion, 52.1%). The weighted mean (SD) age was 45.1 (17.3) years at wave 1 and 48.2 (17.3) years at wave 2. Attempted suicide during the 3 years between wave 1 and wave 2 interviews was self-reported by 222 of 34 653 participants (0.6%). Using survey questions measured at wave 1, the suicide attempt risk model yielded a cross-validated area under the receiver operator characteristic curve of 0.857 with a sensitivity of 85.3% (95% CI, 79.8-89.7) and a specificity of 73.3% (95% CI, 72.8-73.8) at an optimized threshold. The model identified 1.8% of the US population to be at a 10% or greater risk of suicide attempt. The most important risk factors were 3 questions about previous suicidal ideation or behavior; 3 items from the 12-Item Short Form Health Survey, namely feeling downhearted, doing activities less carefully, or accomplishing less because of emotional problems; younger age; lower educational achievement; and recent financial crisis. In this study, after searching through more than 2500 survey questions, several well-known risk factors of suicide attempt were confirmed, such as previous suicidal behaviors and ideation, and new risks were identified, including functional impairment resulting from mental disorders and socioeconomic disadvantage. These results may help guide future clinical assessment and the development of new suicide risk scales.


Acetazolamide fails to decrease pulmonary artery pressure at high altitude in partially acclimatized humans.
In this randomized, double-blind placebo controlled trial our objectives were to determine if acetazolamide is capable of preventing high altitude pulmonary edema (HAPE) in trekkers traveling between 4250 m (Pheriche)\4350 m (Dingboche) and 5000 m (Lobuje) in Nepal; to determine if acetazolamide decreases pulmonary artery systolic pressures (PASP) at high altitude; and to determine if there is an association with PASP and signs and symptoms of HAPE. Participants received either acetazolamide 250 mg PO BID or placebo at Pheriche\Dingboche and were reassessed in Lobuje. The Lake Louise Consensus Criteria were used for the diagnosis of HAPE, and cardiac ultrasonography was used to measure the velocity of tricuspid regurgitation and estimate PASP. Complete measurements were performed on 339 of the 364 subjects (164 in the placebo group, 175 in the acetazolamide group). No cases of HAPE were observed in either study group nor were differences in the signs and symptoms of HAPE found between the two groups. Mean PASP values did not differ significantly between the acetazolamide and placebo groups (31.3 and 32.6 mmHg, respectively). An increasing number of signs and symptoms of HAPE was associated with elevated PASP (p < 0.01). The efficacy of acetazolamide against acute mountain sickness, however, was significant with a 21.9% incidence in the placebo group compared to 10.2 % in the acetazolamide group (p < 0.01). Given the lack of cases of HAPE in either group, we can draw no conclusions about the efficacy of acetazolamide in preventing HAPE, but the absence of effect on PASP suggests that any effect may be minor possibly owing to partial acclimatization during the trek up to 4200 m.


[The dependence of spatial-motor asymmetry on the nature of the reinforcement].
Spatial-motor asymmetry (SMA) of rats goal-directed running was studied in condition of numerous alternative choices in complex T-maze. In condition of free choices SMA in trained rats depended on reinforcement biological significance, volume and disposition scheme. In condition of non-free choices a change was possible of asymmetry of sequence of goal-directed motor acts adequate to situation, i.e. SMA regulation. Examples of the presence of properties of ego- and allocentric systems in SMA are given. The results of experiments testify to SMA dependence on the motivation of running, to rightfulness of SMA interpretation as behavioural tactics and to non-coming of SMA of goal-directed running to a simple rotation of animals.


[Effect of bortezomib on VEGF gene expression of endothelial cell line HMEC-1 and its possible mechanisms].
This study was aimed to investigate the effects of bortezomib on VEGF gene expression of endothelial cell line HMEC-1, and to determine the changes of the transcriptional regulation activity of hypoxia-inducible factor 1 (HIF-1alpha) and expression intensity of Annexin A2, so as to analyze the possible mechanisms of the above expression of VEGF gene. Expression intensity of VEGF gene was determined by real-time quantitative PCR; the relative proliferation activity of cells was assayed by cell count kit CCK-8; the expression intensity of carbonic anhydrase IX (CA IX) gene was detected by RT-PCR; expression of Annexin A2 at gene and protein levels were determined by real-time quantitative PCR and Western blot respectively. The results showed that after being treated by bortezomib with 2.5, 5.0, 10 nmol/L for 12 hours, the expression intensity of VEGF gene of endothelial cell line HMEC-1 was as follows: 0.730 +/- 0.106, 0.673 +/- 0.153, 0.767 +/- 0.090 (as 1.0 was made in 0 nmol/L) (p < 0.05); the proliferation activity of cells was not significantly suppressed by bortezomib in 2.5, 5.0 nmol/L (p > 0.05), while that was significantly suppressed by bortezomib of 10 nmol/L (p = 0.024), The results from RT-PCR showed that expression intensity of CA IX gene was conspicuously down-regulated by bortezomib in different concentrations, which suggested that the transcriptional regulation activity of HIF-1alpha was inhibited by bortezomib. And down-regulated expression of Annexin A2 protein by bortezomib in different concentrations was confirmed by real-time quantitative PCR and Western blot. It is concluded that low doses of bortezomib has no significant inhibition effect on the activity of proteasome. Bortezomib may down-regulate the expression of VEGF gene of endothelial cell through regulating the activity of HIF-1alpha and the expression of Annexin A2.


Influence of the number and distribution of NLS peptides on the photosensitizing activity of multimeric porphyrin-NLS.
Porphyrin-peptide conjugates bearing multiple nuclear localization sequences (NLS) could show increased tumor cell uptake and affinity for nuclear receptors, and consequently increased photodynamic activity. Previous studies suggest that an increase number of NLS might enhance the nuclear uptake of proteins and other macromolecules. We report the syntheses and investigation of a series of multimeric porphyrin-NLS conjugates bearing two, three or four peptides with the minimum sequence PKKKRKV, linked via PEG or 5-carbon linkers, and with different distributions at the porphyrin periphery. Our results show that the tumor cell uptake and phototoxicity of these conjugates is mainly determined by their amphiphilic character, and not the number of NLS residues per molecule, contrary to previous studies. The mono- and di-substituted photosensitizers bearing one or two PEG linkers and up to three peptide sequences were found to be the most phototoxic toward human carcinoma HEp2 cells, while the tetra-NLS conjugates symmetrically substituted around the porphyrin ring accumulated the least within cells and were non-phototoxic. All conjugates localized intracellularly within endosomal vesicles and lysosomes, probably as a result of an endocytic mechanism of uptake; as a consequence no nuclear uptake was detected by fluorescence microscopy.


[The determination of a gross utilization of 15N-lysine in laboratory rats. 1. Experiment with normal intestinal flora (without antibiotic supplement)].
Wistar rats of a live weight of about 100 g were divided into 14 groups (5 animals/group). The rations given supplied the animals with 75%, 100% and 125% lysine, which brought about a moderate growth of the animals of approximately 2 g/animal and day achieved by limited feeding. The 3 lysine levels mentioned could be achieved by lysine supplements (L-lysine-HCl) for the following rations: barley (B), wheat (W), and wheat gluten (WG). For isolated soybean protein (assay protein) (S) the lysine levels 100% and 125% and for soybean meal (SM) the levels 116% and 125% could only be achieved. A control group with whole egg ration (W) (with its natural lysine content of 125% of the requirement) were also tested as comparison. During the 10-day period of the main experiment all 14 rations were supplemented with 0.5 g 15N-lysine (alpha amino group, 95% labelled with 15N). The N balance could only be significantly improved by lysine supplements in the rations B, W and SM with the lysine level of 125%. The biologic value of the protein sources was in rations B and WG also significantly improved by the highest lysine supplement. 15N excess (15N') from the deaminated 15N lysine was excreted with diet B rich in crude fibre mainly in faeces (more than 15% of the intake) and only about 10% in urine. With the diets without native crude fibre the excretion quota changed in favour of urine. The following 15N' amounts in per cent of 15N' intake from lysine were excreted in urine and faeces: B 75 = 31.3, B 100 = 30.9, B 125 = 28.0, W 75 = 24.3, W 100 = 32.2, W 125 = 32.6, GW 75 = 18.3, WG 100 = 24.2, WG 125 = 28.1, S 100 = 39.4, S 125 = 50.4, SM 116 = 34.9, SM 125 = 32.9, W 125 = 19.1. 15N excretion in urine and faeces increased in comparable relations in 6 cases of lysine increase levels only. Gross utilization of lysine can only conditionally be quantified by 15N labelled lysine supplement.


Altered White Matter and microRNA Expression in a Murine Model Related to Williams Syndrome Suggests That miR-34b/c Affects Brain Development via Ptpru and Dcx Modulation.
Williams syndrome (WS) is a multisystem neurodevelopmental disorder caused by a de novo hemizygous deletion of ~26 genes from chromosome 7q11.23, among them the general transcription factor II-I (GTF2I). By studying a novel murine model for the hypersociability phenotype associated with WS, we previously revealed surprising aberrations in myelination and cell differentiation properties in the cortices of mutant mice compared to controls. These mutant mice had selective deletion of Gtf2i in the excitatory neurons of the forebrain. Here, we applied diffusion magnetic resonance imaging and fiber tracking, which showed a reduction in the number of streamlines in limbic outputs such as the fimbria/fornix fibers and the stria terminalis, as well as the corpus callosum of these mutant mice compared to controls. Furthermore, we utilized next-generation sequencing (NGS) analysis of cortical small RNAs' expression (RNA-Seq) levels to identify altered expression of microRNAs (miRNAs), including two from the miR-34 cluster, known to be involved in prominent processes in the developing nervous system. Luciferase reporter assay confirmed the direct binding of miR-34c-5p to the 3'UTR of PTPRU-a gene involved in neural development that was elevated in the cortices of mutant mice relative to controls. Moreover, we found an age-dependent variation in the expression levels of doublecortin (Dcx)-a verified miR-34 target. Thus, we demonstrate the substantial effect a single gene deletion can exert on miRNA regulation and brain structure, and advance our understanding and, hopefully, treatment of WS.


Dark Energy from Violation of Energy Conservation.
In this Letter, we consider the possibility of reconciling metric theories of gravitation with a violation of the conservation of energy-momentum. Under some circumstances, this can be achieved in the context of unimodular gravity, and it leads to the emergence of an effective cosmological constant in Einstein's equation. We specifically investigate two potential sources of energy nonconservation-nonunitary modifications of quantum mechanics and phenomenological models motivated by quantum gravity theories with spacetime discreteness at the Planck scale-and show that such locally negligible phenomena can nevertheless become relevant at the cosmological scale.


Criterion validity and the utility of reactive and proactive aggression: comparisons to attention deficit hyperactivity disorder, oppositional defiant disorder, conduct disorder, and other measures of functioning.
Examined the criterion validity and the utility of Dodge and Coie's (1987) measure of reactive and proactive aggression. Participants were 405 children in kindergarten through 5th grade attending an urban elementary school. Examined criterion validity by testing whether reactive aggression or proactive aggression was significantly correlated with criterion measures of overall impairment as measured by the Impairment Rating Scale (Pelham, Gnagy, et al., 1996), classroom behavior as measured by a frequency count of classroom rule violations, and peer adjustment as measured by teacher ratings of peer behavior on the Pittsburgh Modified IOWA Conners (Pelham, Milich, Murphy, & Murphy, 1989). Examined utility by testing whether reactive aggression or proactive aggression was significantly correlated with criterion measures after controlling for each other and after controlling for attention deficit hyperactivity disorder, oppositional defiant disorder, and conduct disorder as measured by the Disruptive Behavior Disorder Rating Scales (Pelham, Gnagy, Greenslade, & Milich, 1992). Results showed good evidence of criterion validity for both reactive and proactive aggression, but mixed evidence for their utility.


Radiolucent inflammatory implant periapical lesions: a review of the literature.
To discuss the terminology, etiopathogenesis, and treatment of radiolucent inflammatory implant periapical lesions. An electronic search for relevant articles published in the English literature in the PubMed database. Bacterial contamination of the apical portion of the implant either from a preexisting dental periapical infection or from a periapical lesion of endodontic origin of an adjacent tooth is the probable causative factor. Aseptic bone necrosis owing to overheating of the bone during preparation of osteotomies, or compression of the bone at the apex of the implant owing to excessive tightening, may also play a role. The histopathological features are of a mixed inflammatory cell infiltrate on a background of granulation tissue consistent with either a granuloma or an abscess as may be found at the apex of a nonvital tooth. Treatment consists of immediate and aggressive surgical debridement, chemical detoxification of the apical portion of the exposed implant surface, and systemic antibiotics with or without a bone regenerative procedure. A radiolucent inflammatory implant periapical lesion is analogous to either a granuloma or an abscess as may be found at the apex of a nonvital tooth.


Axotomy-induced miR-21 promotes axon growth in adult dorsal root ganglion neurons.
Following injury, dorsal root ganglion (DRG) neurons undergo transcriptional changes so as to adopt phenotypic changes that promote cell survival and axonal regeneration. Here we used a microarray approach to profile changes in a population of small noncoding RNAs known as microRNAs (miRNAs) in the L4 and L5 DRG following sciatic nerve transection. Results showed that 20 miRNA transcripts displayed a significant change in expression levels, with 8 miRNAs transcripts being altered by more than 1.5-fold. Using quantitative reverse transcription PCR, we demonstrated that one of these miRNAs, miR-21, was upregulated by 7-fold in the DRG at 7 days post-axotomy. In dissociated adult rat DRG neurons lentiviral vector-mediated overexpression of miR-21 promoted neurite outgrowth on a reduced laminin substrate. miR-21 directly downregulated expression of Sprouty2 protein, as confirmed by Western blot analysis and 3' untranslated region (UTR) luciferase assays. Our data show that miR-21 is an axotomy-induced miRNA that enhances axon growth, and suggest that miRNAs are important players in regulating growth pathways following peripheral nerve injury.


Evaluation of Streptomyces spp. for their plant-growth-promotion traits in rice.
Five strains of Streptomyces (CAI-17, CAI-68, CAI-78, KAI-26, and KAI-27) were previously reported to have potential for charcoal rot control and plant growth promotion (PGP) in sorghum. In this study, those 5 Streptomyces strains were characterized for their enzymatic activities and evaluated for their PGP capabilities on rice. All the Streptomyces strains were able to produce lipase and β-1,3-glucanase; grew in NaCl (up to 8%), at pH 5-13, and at temperatures 20-40 °C; and were resistant to ampicillin, sensitive to nalidixic acid, and highly sensitive to chloramphenicol, kanamycin, streptomycin, and tetracycline. They were highly tolerant to the fungicide bavistin but were highly sensitive to benlate, benomyl, and radonil. When evaluated on rice in the field, Streptomyces significantly enhanced tiller and panicle numbers, stover and grain yields, dry matter, root length, volume and dry weight, compared with the control. In the rhizosphere at harvest, microbial biomass carbon and nitrogen, dehydrogenase activity, total nitrogen, available phosphorus, and % organic carbon were also found significantly higher in Streptomyces-treated plots than in the control plots. This study further confirms that the selected Streptomyces have PGP activities.


Proteomic analysis of cervical vaginal fluid proteins among women in recurrent preterm labor.
Proteomic analysis of four cervical-vaginal fluid (CVF) proteins to identify biomarkers of recurrent preterm birth (rPTB) in at-risk women prior to onset of preterm labor. Nested case control study from 2007 to 2011 of women with prior spontaneous preterm birth(s) (PTB) who underwent serial CVF sampling. Mass spectrometry analysis was used and ELISA analysis was performed to validate candidates. 108 patients were enrolled and 10 cases and 20 gestational age matched controls were analyzed after exclusions. Of 748 CVF proteins identified, 72 had statistically significant (p < 0.05) expression differences and 38 were highly differentially expressed (p < 0.01). Four candidate proteins were abundant and involved in immune/inflammatory response, but ELISA analysis did not confirm altered expression patterns. The lack of confirmation of potential biomarkers identified by mass spectrometry and ELISA demonstrates the challenges of validating PTB biomarkers and suggests that a panel of biomarkers would improve the predictive value of CVF testing.


Isolated knee monoarthritis heralding resectable non-small-cell lung cancer. A paraneoplastic syndrome not previously described.
To describe isolated knee monoarthritis as a paraneoplastic syndrome heralding non-small cell lung cancer (NSCLC), and to discuss its clinical characteristics. Clinical records of all consecutive, new outpatients with isolated knee monoarthritis observed from January 2000 to December 2005 were reviewed. A systematic review of Medline and Cochrane Library databases was performed to identify English-language articles related to rheumatological paraneoplastic syndromes associated with NSCLC. Over 6 years, 6654 new outpatients with different rheumatic disorders were observed. Of these, 296 (4.4%) presented with isolated monoarthritis of the knee. In five out of 296 patients (1.7%) this feature represented the initial manifestation of NSCLC. All five patients were middle-aged men, with a long history of heavy cigarette smoking, who had a non-erosive, isolated knee monoarthritis, with mild articular fluid collection of non-inflammatory type. NSCLC was resectable in all patients, and knee monoarthritis remitted with no relapse confirming its paraneoplastic nature. All five patients are in good condition after a median follow up of 41 months. The literature review revealed that paraneoplastic knee monoarthritis has not previously been reported. Knee monoarthritis may in some cases represent a paraneoplastic syndrome heralding NSCLC at an early stage.


[Scapholunate dissociation. Results of ligament suture and ligament-plasty].
The diagnosis of scapholunate ligament rupture is frequently missed. Untreated, it will lead to carpal collapse and arthrosis of the wrist, and ultimately to scapholunate advanced collapse (SLAC-wrist). We report the results of 20 patients treated either by direct suture or reconstruction of the ligament with a follow-up of three to five years after surgery. The operation was performed between 0.5 and 120 months (suture 2.6 months, reconstruction 28 months) after the original trauma. The results of direct repair were superior to ligament reconstruction. In our experience, direct ligament repair was only possible up to four to six months after trauma, this study emphasizes the importance of early diagnosis and treatment by anatomical reposition of scapholunate ligament rupture.


Spexin and a Novel Cichlid-Specific Spexin Paralog Both Inhibit FSH and LH Through a Specific Galanin Receptor (Galr2b) in Tilapia.
Spexin (SPX) is a 14 amino acid peptide hormone that has pleiotropic functions across vertebrates, one of which is involvement in the brain-pituitary-gonad axis of fish. SPX(1) has been identified in each class of vertebrates, and a second SPX (named SPX2) has been found in some non-mammalian species. We have cloned two spexin paralogs, designated as Spx1a and Spx1b, from Nile tilapia (Oreochromis niloticus) that have varying tissue distribution patterns. Spx1b is a novel peptide only identified in cichlid fish, and is more closely related to Spx1 than Spx2 homologs as supported by phylogenetic, synteny, and functional analyses. Kisspeptin, Spx, and galanin (Gal) peptides and their corresponding kiss receptors and Gal receptors (Galrs), respectively, are evolutionarily related. Cloning of six tilapia Galrs (Galr1a, Galr1b, Galr2a, Galr2b, Galr type 1, and Galr type 2) and subsequent in vitro second-messenger reporter assays for Gαs, Gαq, and Gαi suggests that Gal and Spx activate Galr1a/Galr2a and Galr2b, respectively. A decrease in plasma follicle stimulating hormone and luteinizing hormone concentrations was observed with injections of Spx1a or Spx1b in vivo. Additionally, application of Spx1a and Spx1b to pituitary slices decreased the firing rate of LH cells, suggesting that the peptides can act directly at the level of the pituitary. These data collectively suggest an inhibitory mechanism of action against the secretion of gonadotropins for a traditional and a novel spexin paralog in cichlid species.


Predicting the no-reflow phenomenon following successful percutaneous coronary intervention.
In the setting of acute myocardial infarction, early and adequate reopening of an infarct-related artery is not necessarily followed by a complete restoration of myocardial perfusion. This condition is usually defined as 'no-reflow'. The pathophysiology of no-reflow is multifactorial since extravascular compression, microvascular vasoconstriction, embolization during percutaneous coronary intervention, and platelet and neutrophil aggregates are involved. In the clinical arena, angiographic findings and easily available clinical parameters can predict the risk of no-reflow. More recently, several studies have demonstrated that biomarkers, especially those related to the pathogenetic components of no-reflow, could also have a prognostic role in the prediction and in the full understanding of the multiple mechanisms of this phenomenon. Thus, in this article, we investigate the role of several biomarkers on admission in predicting the occurrence of no-reflow following successful percutaneous coronary intervention.


Neuromuscular Development and Disease: Learning From in vitro and in vivo Models.
The neuromuscular junction (NMJ) is a specialized cholinergic synaptic interface between a motor neuron and a skeletal muscle fiber that translates presynaptic electrical impulses into motor function. NMJ formation and maintenance require tightly regulated signaling and cellular communication among motor neurons, myogenic cells, and Schwann cells. Neuromuscular diseases (NMDs) can result in loss of NMJ function and motor input leading to paralysis or even death. Although small animal models have been instrumental in advancing our understanding of the NMJ structure and function, the complexities of studying this multi-tissue system in vivo and poor clinical outcomes of candidate therapies developed in small animal models has driven the need for in vitro models of functional human NMJ to complement animal studies. In this review, we discuss prevailing models of NMDs and highlight the current progress and ongoing challenges in developing human iPSC-derived (hiPSC) 3D cell culture models of functional NMJs. We first review in vivo development of motor neurons, skeletal muscle, Schwann cells, and the NMJ alongside current methods for directing the differentiation of relevant cell types from hiPSCs. We further compare the efficacy of modeling NMDs in animals and human cell culture systems in the context of five NMDs: amyotrophic lateral sclerosis, myasthenia gravis, Duchenne muscular dystrophy, myotonic dystrophy, and Pompe disease. Finally, we discuss further work necessary for hiPSC-derived NMJ models to function as effective personalized NMD platforms.


Managing genital chlamydia trachomatis infection in Scotland: targeted opportunistic testing or a screening programme?
To compare two models of testing for genital chlamydia trachomatis infection: opportunistic testing for genital chlamydia trachomatis at sexual health related consultations, as advocated in the SIGN guideline on Chlamydia trachomatis, or the introduction of a screening programme. Calculation of estimates of numbers of cases found in one Health Board Area using the two different approaches. Review of the literature on chlamydia with reference to the UK National Screening Committee criteria for appraisal of a screening programme. Lothian Health Board Area. The evidence suggests that a screening programme for chlamydia has population benefits and is cost effective compared to a no-screening approach. However, current understanding of the natural history of chlamydial infection and the effect of treatment is inadequate to fully inform individual women regarding the benefit of testing. Full implementation of the SIGN guideline on Chlamydia trachomatis in Lothian would detect at least 600 new cases of chlamydia whereas a screening programme would detect over 2000 new cases in the first year. Opportunistic testing at sexual health related consultations does not equate with a screening programme. Although a screening programme would detect more cases of chlamydia trachomatis its cost effectiveness compared to opportunistic testing is unproven. Introducing a screening programme has individual psychological and broader social and ethical implications and must not occur by default.


MHC class I-specific inhibitory receptors and their ligands structure diverse human NK-cell repertoires toward a balance of missing self-response.
Variegated expression of 6 inhibitory HLA class I-specific receptors on primary NK cells was studied using high-dimension flow cytometry in 58 humans to understand the structure and function of NK-cell repertoires. Sixty-four subsets expressing all possible receptor com-binations were present in each repertoire, and the frequency of receptor-null cells varied among the donors. Enhancement in missing-self response between NK subsets varied substantially where subset responses were defined by donor KIR/HLA allotypes, reflecting the differences in interaction between inhibitory receptors and their ligands. This contrasted to the enhancement conferred by NKG2A, which was constant and of intermediate strength. We infer a mechanism that modulates frequencies of the NK subsets displaying diverse levels of missing-self response, a system that reduces the presence of KIR-expressing subsets that display either too strong or too weak a response and effectively replaces them with NKG2A-expressing cells in the repertoire. Through this high-resolution analysis of inhibitory receptor expression, 5 types of NK-cell repertoire were defined by their content of NKG2A(+)/NKG2A(-) cells, frequency of receptor-null cells, and degree of KIR receptor coexpression. The analyses provide new perspective on how personalized human NK-cell repertoires are structured.


The cytotoxic, apoptotic and oxidative effects of carbonic anhydrase IX inhibitor on colorectal cancer cells.
Colorectal cancer (CRC) is the third most common tumor, malignant and has developed one of the main reasons of cancer mortality. According to studies conducted recently; carbonic anhydrase 9 (CAIX) is an especially attractive target for cancer therapy, in part since it is limited way expressed in normal tissues on the other hand in a wide variety of solid neoplasia are overexpressed. The aim of this study was to appreciate the effects of CAIX inhibitor, namely novel synthesized sulfonamide derivative (H-4i) with high affinity for CAIX, in CAIX-positive human colorectal cancer cell (HT-29) and CAIX-negative human normal embryonic kidney cell line (HEK-293). For this reason, we planned to investigate apoptotic, cytotoxic and oxidative stress activity of H-4i on HT-29 and HEK-293 cell lines. Cell viability determined by WST-1 assay afterwards IC50 values, apoptosis and cell cycle induction measured by flow cytometric analysis, intracellular free radical induction performed by reactive oxygen species (ROS) analyses. The IC50 value of the sulfonamide derivative compound was found to be very low, especially in HT-29 cells, when compared to human normal cells. This research found that H-4i significantly increased cytotoxicity and ROS production, caused significant signs of apoptosis level. High level of ROS and apoptosis lead to arrest the cell cycle and reduce cell survival. The most obvious finding to emerge from the analysis that novel synthesized sulfonamide derivative H-4i is effective on HT-29 more than HEK-293. Therefore, novel derivative H-4i might be used as an anti-cancer potential compound on CRC.


Field drought conditions impact yield but not nutritional quality of the seed in common bean (Phaseolus vulgaris L.).
Drought substantially limits seed yield of common bean (Phaseolus vulgaris L.) in the tropics. Understanding the interaction of drought on yield and the nutrient concentration of the seed is vital in order to supply nutrition to the millions of consumers who rely on common bean as a staple crop. Nevertheless, the impact of drought on common bean for both yield and nutrient concentration has not yet been concurrently investigated in a field environment. Using 10 bred lines developed by CIAT and its partners for their improved adaptation to drought and phosphorus deficiency, this study characterised the impact of drought on yield and nutrient concentration for leaf and seed tissue of common bean grown in the field. Drought significantly reduced leaf area (by ~50%), harvest index (by ~60%), yield (by ~70%), seed weight (by ~25%) and enriched carbon isotope abundance (δ13C) in the seed. Within the soluble leaf fraction, drought significantly decreased the concentration of mineral nutrients and amino acids, whereas no negative effect on the concentration of nutrients and amino acids was detected within the seed. Genotypic variation in nutrient concentration in both the leaf and seed tissue was identified and should be explored further to identify traits that may confer tolerance to abiotic stress.


Subpellicular microtubules of Euplotes eurystomus: their geometry relative to cell form, surface contours and ciliary organelles.
There is a layer of microtubules (MT) beneath the innermost pellicular membrane of the ciliated protozoan Euplotes eurystomus. These MT have been revealed for scanning electron microscopic study by chemical dissection techniques. In much of the body of this ciliate, these MT are oriented parallel to its long axis. Those directed towards cirri, which are complex ciliary structures of the ventral surface, either abut or bend around the cirral base. MT adjacent to or closely associated with the ciliary feeding structures (membranelles of the adoral zone of membranelles or AZM) are oriented parallel to the long axis of the AZM. Some of the MT within the oral cavity have quite complex paths. The various orientations of these subpellicular MT are discussed and evaluated for hypothetical functions of cytoskeletal support, cell shaping and organelle movement. Each of these roles is considered to be theoretically possible.


[Evolution of the prevalence of intestinal parasitosis among schoolchildren in Caxias do Sul, RS].
Reports on the prevalence of intestinal parasitosis in Brazil have been local in nature, with descriptions of different populations, which makes comprehensive diagnosis difficult. With the aim of studying the variation in the prevalence of intestinal parasitosis among schoolchildren in Caxias do Sul, Rio Grande do Sul, over a 35-year period, 9,787 parasitological stool tests that had been performed using centrifugal sedimentation were evaluated. There were positive results from 5,655 samples (58%), and the most prevalent infestations were of Ascaris lumbricoides (47%), Trichuris trichiura (36%), Enterobius vermicularis (8%) and the protozoa Giardia lamblia (24%) and Entamoeba coli (20%). The overall prevalence diminished from 89% to 37%, indicating an average decrease of 1.4% per year. Reductions in prevalence were observed for Ascaris lumbricoides (61 to 26%) and Trichuris trichiura (38 to 18%). No significant change was observed for Giardia lamblia. The prevalence of Entamoeba coli increased from 29 to 46%. The decreases in helminth prevalence were probably due to infrastructure improvements and educational actions undertaken in schools.


Desyndactylization: a unique case report.
Syndactylism is a congenital or acquired anomaly in which there is persistence of webbing between adjacent digits. It occurs in 1:2000 to 1:2500 live births, three times more commonly in males, and most often between the middle and third fingers. In the foot, syndactyly involves the second and third toes most frequently. Generally, this is thought of as a cosmetic condition when associated with the toes of the foot, as this usually causes little functional loss. The authors present a unique case report of syndactyly of the hallux and second toe in a 66-year-old black female and its surgical repair for painful hammertoe deformity.


Rostral raphe involvement in Lewy body dementia and multiple system atrophy.
Depression is a feature of both Lewy body disorders and multiple system atrophy (MSA). Since serotonergic neurons of the rostral raphe have been implicated in depression, we sought to determine whether there is a differential involvement of these neurons in cases with clinically diagnosed dementia with Lewy bodies (DLB) or MSA. We studied the brainstem obtained at autopsy from fourteen patients with diagnosis of DLB and pathological limbic or neocortical stage Lewy body disease, 13 patients with clinical and neuropathological diagnosis of MSA, and 12 controls with no history of neurologic disease. The clinical features of these patients were analyzed retrospectively by reviewing their medical records. Serial sections were immunostained for tryptophan hydroxylase (TrOH) and alpha-synuclein and cell counts were performed in the dorsal raphe (DR), median raphe (MR) and medullary raphe nuclei. There was loss of serotonergic cells in both the DR and MR in DLB compared to control cases: For the DR, the number of cells/section were 53 +/- 6 in DLB versus 159 +/- 13 (P < 0.001) respectively, and for the MR 70 +/- 11 in DLB versus 173 +/- 23 (P < 0.001) respectively. In contrast, these cells were relatively preserved in MSA. The caudal raphe groups were affected both in MSA and in DLB. There is a differential involvement of raphe neurons in DLB and MSA. Although loss of rostral raphe neurons may contribute to depression in DLB, this appears to be less likely in MSA. Factors other than the neurochemical phenotype determine neuronal vulnerability in MSA.


Medical signs and symptoms associated with disability, pain, and psychosocial adjustment in systemic sclerosis.
To examine physician-assessed medical signs and patient-reported medical symptoms as correlates of 3 quality of life (QOL) outcomes in patients with systemic sclerosis (SSc): disability, pain, and psychosocial adjustment. One hundred fourteen patients with SSc underwent a comprehensive clinical examination including determination of skin thickening [Modified Rodnan Skin Score (MRSS)]. Patients reported current symptoms and completed standardized questionnaires assessing disability and pain (Health Assessment Questionnaire) and psychosocial adjustment (Psychosocial Adjustment to Illness Scale). Regression analysis was used to examine physician-determined and patient-reported correlates of the 3 outcomes. MRSS was a significant correlate of all outcomes, although it explained only a small amount of the variance in psychosocial adjustment. Patient-reported postprandial bloating was the strongest correlate of psychosocial adjustment, explaining more than twice as much variance as MRSS. After accounting for MRSS, patient-reported dependent edema significantly correlated with all outcomes. For disability, significant correlates were physician-determined joint tenderness and number of tender points, and patient-reported joint pain on motion, joint contracture, extremity ulcers other than digital, and dyspnea. Patient-reported joint tenderness was significantly associated with pain. Regression analysis supported a model in which disability and pain mediated the relationship between MRSS and psychosocial adjustment. Skin score is strongly associated with disability and pain, but only weakly associated with psychosocial adjustment. Dependent edema has negative implications across quality-of-life outcomes. Disability and pain mediate the relationship between disease severity and psychosocial adjustment to disease. Assessment (including self-report of patient symptoms) of specific medical signs and symptoms may indicate SSc patients experiencing diminished QOL.


Survey of leafhopper species in almond orchards infected with almond witches'-broom phytoplasma in Lebanon.
Leafhoppers (Hemiptera: Auchenorrhyncha: Cicadellidae) account for more than 80% of all "Auchenorrhynchous" vectors that transmit phytoplasmas. The leafhopper populations in two almond witches'-broom phytoplasma (AlmWB) infected sites: Tanboureet (south of Lebanon) and Bourj El Yahoudieh (north of Lebanon) were surveyed using yellow sticky traps. The survey revealed that the most abundant species was Asymmetrasca decedens, which represented 82.4% of all the leafhoppers sampled. Potential phytoplasma vectors in members of the subfamilies Aphrodinae, Deltocephalinae, and Megophthalminae were present in very low numbers including: Aphrodes makarovi, Cicadulina bipunctella, Euscelidius mundus, Fieberiella macchiae, Allygus theryi, Circulifer haematoceps, Neoaliturus transversalis, and Megophthalmus scabripennis. Allygus theryi (Horváth) (Deltocephalinae) was reported for the first time in Lebanon. Nested PCR analysis and sequencing showed that Asymmetrasca decedens, Empoasca decipiens, Fieberiella macchiae, Euscelidius mundus, Thamnottetix seclusis, Balclutha sp., Lylatina inexpectata, Allygus sp., and Annoplotettix danutae were nine potential carriers of AlmWB phytoplasma. Although the detection of phytoplasmas in an insect does not prove a definite vector relationship, the technique is useful in narrowing the search for potential vectors. The importance of this information for management of AlmWB is discussed.


A unified approach to noise removal, image enhancement, and shape recovery.
We present a unified approach to noise removal, image enhancement, and shape recovery in images. The underlying approach relies on the level set formulation of the curve and surface motion, which leads to a class of PDE-based algorithms. Beginning with an image, the first stage of this approach removes noise and enhances the image by evolving the image under flow controlled by min/max curvature and by the mean curvature. This stage is applicable to both salt-and-pepper grey-scale noise and full-image continuous noise present in black and white images, grey-scale images, texture images, and color images. The noise removal/enhancement schemes applied in this stage contain only one enhancement parameter, which in most cases is automatically chosen. The other key advantage of our approach is that a stopping criteria is automatically picked from the image; continued application of the scheme produces no further change. The second stage of our approach is the shape recovery of a desired object; we again exploit the level set approach to evolve an initial curve/surface toward the desired boundary, driven by an image-dependent speed function that automatically stops at the desired boundary.


Dyssynergic Defecation in Inflammatory Bowel Disease: A Systematic Review and Meta-Analysis.
Inflammatory bowel disease (IBD) patients often continue to experience nonspecific gastrointestinal symptoms despite quiescent disease. Unlike non-IBD patients, IBD patients with dyssynergic defecation (DD) may present with various symptoms such as diarrhea, fecal incontinence, constipation, and rectal discomfort. Despite its importance and treatability, DD in IBD patients is not well recognized in practice. We conducted a systematic review and meta-analysis on the prevalence, diagnosis, and management of DD in IBD patients with ongoing defecatory symptoms. We searched MEDLINE, EMBASE, and Cochrane Database of Systematic Reviews (from 1966 through February 2017) to identify relevant studies on the prevalence, diagnostic methods, or management of DD in IBD patients with and without ileal pouch-anal anastomoses (IPAAs). A random effects model was used to calculate the pooled estimates with 95% confidence intervals (CIs). Heterogeneity was assessed with I2 statistics, Cochran Q statistic, and sensitivity analyses. Seven studies (n = 442) were included. In patients with ongoing defecatory symptoms, the prevalence of DD without IPAA ranged from 45% to 97%, and in patients with IPAA, it ranged from 25% to 75%. The prevalence of DD in IPAA patients with and without pouchitis ranged from 17% to 67% and 29% to 50%, respectively. The pooled response rate to biofeedback therapy in patients without IPAA was 70% (95% CI, 55%-84%; I2 = 95%; P < 0.01), and it was 86% (95% CI, 67%-98%; I2 = 61%; P = 0.05) in those with IPAA. Despite limited data, the current literature suggests that DD is highly prevalent in active or quiescent IBD patients with ongoing defecatory symptoms and is responsive to biofeedback therapy. Although more studies are needed, DD should be considered in IBD patients with persistent defecatory symptoms.


Sympathetic discharge to mesenteric organs and the liver. Evidence for substantial mesenteric organ norepinephrine spillover.
This study using sampling of blood from the portal vein, in addition to arterial and hepatic sites, to estimate separately spillovers of norepinephrine from mesenteric organs and the liver in seven patients undergoing upper abdominal surgery. Conventional measurements in arterial and hepatic venous plasma provided a measure of net hepatomesenteric NE spillover (403 pmol/ml) that indicated a 13% contribution of these organs to total body spillover of NE into systemic plasma (3,071+/-518 pmol/min). The net hepatomesenteric spillover of NE into systemic plasma was much lower than the spillover of NE from mesenteric organs into portal venous plasma (1,684+/-418 pmol/min). This and the hepatic spillover of NE into systemic plasma (212+/-72 pmol/min) indicated a considerable combined spillover of NE from hepatomesenteric organs (1,896+/-455 pmol/min). The sum of the latter estimate with the difference between total body and net hepatomesenteric NE spillovers provided an adjusted total body spillover of NE into both systemic and portal venous plasma (4,564+/-902 pmol/min). Mesenteric organs made a 37% contribution, and the liver made a 5% contribution to the adjusted total body spillover of NE. Thus, a substantial proportion of total body sympathetic outflow is directed towards mesenteric organs; this is obscured by efficient hepatic extraction of NE (86+/-6%) when measurements are restricted to arterial and hepatic venous plasma.


Assessing the Impacts of Creating Active Schools on Organisational Culture for Physical Activity.
National and international guidance recommends whole-school approaches to physical activity, but there are few studies assessing their effectiveness, especially at an organisational level. This study assesses the impact of the Creating Active School's (CAS) programme on organisational changes to physical activity provision. In-school CAS leads completed a 77-item questionnaire assessing school-level organisational change. The questionnaire comprised 19 domains aligned with the CAS framework and COM-B model of behaviour change. Wilcoxon Signed Rank Tests assessed the pre-to-nine-month change. >70% of schools (n = 53) pre-CAS had inadequate whole-school physical activity provision. After nine months (n = 32), CAS had a significant positive effect on organisational physical activity. The positive change was observed for: whole-school culture and ethos, teachers and wider school staff, academic lessons, physical education (PE) lessons, commute to/from school and stakeholder behaviour. This study provides preliminary evidence that CAS is a viable model to facilitate system-level change for physical activity in schools located within deprived areas of a multi-ethnic city. To confirm the results, future studies are required which adopt controlled designs combined with a holistic understanding of implementation determinants and underlying mechanisms.


Nano- and macroscale structural and mechanical properties of in situ synthesized bacterial cellulose/PEO-b-PPO-b-PEO biocomposites.
Highly transparent biocomposite based on bacterial cellulose (BC) mat modified with poly(ethylene oxide-b-propylene oxide-b-ethylene oxide) block copolymer (EPE) were fabricated in situ during biosynthesis of bacterial cellulose in a static culture from Gluconacetobacter xylinum. The effect of the addition to the culture medium of water-soluble EPE block copolymer on structure, morphology, crystallinity, and final properties of the novel biocomposites was investigated at nano- and macroscale. High compatibility between components was confirmed by ATR-FTIR indicating hydrogen bond formation between the OH group of BC and the PEO block of EPE block copolymer. Structural properties of EPE/BC biocomposites showed a strong effect of EPE block copolymer on the morphology of the BC mats. Thus, the increase of the EPE block copolymer content lead to the generation of spherulites of PEO block, clearly visualized using AFM and MO technique, changing crystallinity of the final EPE/BC biocomposites investigated by XRD. Generally, EPE/BC biocomposites maintain thermal stability and mechanical properties of the BC mat being 1 wt % EPE/BC biocomposite material with the best properties. Biosynthesis of EPE/BC composites open new strategy to the utilization of water-soluble block copolymers in the preparation of BC mat based biocomposites with tunable properties.


Understanding and mitigating the toxicity of cadmium to the anaerobic fermentation of waste activated sludge.
Cadmium (Cd) is present in significant levels in waste activated sludge, but its potential toxicities on anaerobic fermentation of sludge remain largely unknown. This work therefore aims to provide such support. Experimental results showed that the impact of Cd on short-chain fatty acids (SCFA) production from sludge anaerobic fermentation was dose-dependent. The presence of environmentally relevant level of Cd (e.g., 0.1 mg/g VSS) enhanced SCFA production by 10.6%, but 10 mg/g VSS of Cd caused 68.1% of inhibition. Mechanism exploration revealed that although all levels of Cd did not cause extra leakage of intracellular substrates, 0.1 mg/g VSS Cd increased the contents of both soluble and loosely-bound extracellular polymeric substances (EPS), thereby benefitting sludge solubilization. On the contrary, 10 mg/g VSS Cd decreased the levels of all EPS layers, which reduced the content of soluble substrates. It was also found that 0.1 mg/g VSS Cd benefited both the hydrolysis and acidogenesis but 10 mg/g VSS Cd inhibited all the hydrolysis, acidogenesis, and methanogenesis processes. Further investigations with microbial community and enzyme analysis showed that the pertinent presence of Cd enhanced the activities of protease, acetate kinase, and oxaloacetate transcarboxylase whereas 10 mg/g VSS Cd decreased the microbial diversity, the abundances of functional microbes, and the activities of key enzymes. Finally, one strategy that could effectively mitigate the adverse impact of high Cd levels on SCFA production was proposed and examined. This work provides insights into Cd-present sludge fermentation systems, and the findings obtained may guide engineers to manipulate sludge treatment systems in the future.


Statins Effects on Blood Clotting: A Review.
Statins are powerful lipid-lowering drugs that inhibit cholesterol biosynthesis via downregulation of hydroxymethylglutaryl coenzyme-A reductase, which are largely used in patients with or at risk of cardiovascular disease. Available data on thromboembolic disease include primary and secondary prevention as well as bleeding and mortality rates in statin users during anticoagulation for VTE. Experimental studies indicate that statins alter blood clotting at various levels. Statins produce anticoagulant effects via downregulation of tissue factor expression and enhanced endothelial thrombomodulin expression resulting in reduced thrombin generation. Statins impair fibrinogen cleavage and reduce thrombin generation. A reduction of factor V and factor XIII activation has been observed in patients treated with statins. It is postulated that the mechanisms involved are downregulation of factor V and activated factor V, modulation of the protein C pathway and alteration of the tissue factor pathway inhibitor. Clinical and experimental studies have shown that statins exert antiplatelet effects through early and delayed inhibition of platelet activation, adhesion and aggregation. It has been postulated that statin-induced anticoagulant effects can explain, at least partially, a reduction in primary and secondary VTE and death. Evidence supporting the use of statins for prevention of arterial thrombosis-related cardiovascular events is robust, but their role in VTE remains to be further elucidated. In this review, we present biological evidence and experimental data supporting the ability of statins to directly interfere with the clotting system.


Therapeutic Potential of Exogenous Ketone Supplement Induced Ketosis in the Treatment of Psychiatric Disorders: Review of Current Literature.
Globally, psychiatric disorders, such as anxiety disorder, bipolar disorder, schizophrenia, depression, autism spectrum disorder, and attention-deficit/hyperactivity disorder (ADHD) are becoming more prevalent. Although the exact pathological alterations are not yet clear, recent studies have demonstrated that widespread changes of very complex metabolic pathways may partially underlie the pathophysiology of many psychiatric diseases. Thus, more attention should be directed to metabolic-based therapeutic interventions in the treatment of psychiatric disorders. Emerging evidence from numerous studies suggests that administration of exogenous ketone supplements, such as ketone salts or ketone esters, generates rapid and sustained nutritional ketosis and metabolic changes, which may evoke potential therapeutic effects in cases of central nervous system (CNS) disorders, including psychiatric diseases. Therefore, the aim of this review is to summarize the current information on ketone supplementation as a potential therapeutic tool for psychiatric disorders. Ketone supplementation elevates blood levels of the ketone bodies: D-β-hydroxybutyrate (βHB), acetoacetate (AcAc), and acetone. These compounds, either directly or indirectly, beneficially affect the mitochondria, glycolysis, neurotransmitter levels, activity of free fatty acid receptor 3 (FFAR3), hydroxycarboxylic acid receptor 2 (HCAR2), and histone deacetylase, as well as functioning of NOD-like receptor pyrin domain 3 (NLRP3) inflammasome and mitochondrial uncoupling protein (UCP) expression. The result of downstream cellular and molecular changes is a reduction in the pathophysiology associated with various psychiatric disorders. We conclude that supplement-induced nutritional ketosis leads to metabolic changes and improvements, for example, in mitochondrial function and inflammatory processes, and suggest that development of specific adjunctive ketogenic protocols for psychiatric diseases should be actively pursued.


Transcriptome-wide analyses of adipose tissue in outbred rats reveal genetic regulatory mechanisms relevant for human obesity.
Transcriptomic analysis in metabolically active tissues allows a systems genetics approach to identify causal genes and networks involved in metabolic disease. Outbred heterogeneous stock (HS) rats are used for genetic mapping of complex traits, but to-date, a systems genetics analysis of metabolic tissues has not been done. We investigated whether adiposity-associated genes and gene coexpression networks in outbred heterogeneous stock (HS) rats overlap those found in humans. We analyzed RNAseq data from adipose tissue of 415 male HS rats, correlated these transcripts with body weight (BW) and compared transcriptome signatures to two human cohorts: the "African American Genetics of Metabolism and Expression" and "Metabolic Syndrome in Men." We used weighted gene coexpression network analysis to identify adiposity-associated gene networks and mediation analysis to identify genes under genetic control whose expression drives adiposity. We identified 554 orthologous "consensus genes" whose expression correlates with BW in the rat and with body mass index (BMI) in both human cohorts. Consensus genes fell within eight coexpressed networks and were enriched for genes involved in immune system function, cell growth, extracellular matrix organization, and lipid metabolic processes. We identified 19 consensus genes for which genetic variation may influence BW via their expression, including those involved in lipolysis (e.g., Hcar1), inflammation (e.g., Rgs1), adipogenesis (e.g., Tmem120b), or no previously known role in obesity (e.g., St14 and Ms4a6a). Strong concordance between HS rat and human BW/BMI associated transcripts demonstrates translational utility of the rat model, while identification of novel genes expands our knowledge of the genetics underlying obesity.


Exploring the Pre-exposure Prophylaxis (PrEP) Health Care Experiences Among Men Who Have Sex With Men (MSM) Who Live in Rural Areas of the Midwest.
Most pre-exposure prophylaxis (PrEP) research samples men who have sex with men (MSM) who live in metropolitan cities. There is a limited understanding of the PrEP experiences among rural MSM. Thirty-four semi-structured interviews were conducted to explore the PrEP health care experiences among 34 rural Midwestern MSM. Of the 34 participants, 23 obtained PrEP from their non-primary care provider (PCP). Three themes were present: (1) PrEP is unavailable in rural areas, (2) PrEP is inaccessible in rural areas due to PCPs being unwilling to prescribe PrEP, and (3) PrEP services are unamicable in rural areas due to stigmatizing attitudes and behaviors of PCPs. It is important to address PrEP care continuum factors in all settings; however, these factors are often exacerbated in rural areas due to social determinants. Without further research and programming, organizational and social determinants may contribute to lower rates of PrEP outcomes in rural areas.


Substrate suppression of thermal roughness in stacked supported bilayers.
We have fabricated a stack of five 1,2-dipalmitoyl-sn-3-phosphatidylethanolamine (DPPE) bilayers supported on a polished silicon substrate in excess water. The density profile of these stacks normal to the substrate was obtained through analysis of x-ray reflectivity. Near the substrate, we find the layer roughness and repeat spacing are both significantly smaller than values found in bulk multilayer systems. The reduced spacing and roughness result from suppression of lateral fluctuations due to the flat substrate boundary. The layer spacing decrease then occurs due to reduced Helfrich repulsion.


Physical mapping and InDel marker development for the restorer gene Rf2 in cytoplasmic male sterile CMS-D8 cotton.
Cytoplasmic male sterile (CMS) with cytoplasm from Gossypium Trilobum (D8) fails to produce functional pollen. It is useful for commercial hybrid cotton seed production. The restore line of CMS-D8 containing Rf2 gene can restore the fertility of the corresponding sterile line. This study combined the whole genome resequencing bulked segregant analysis (BSA) with high-throughput SNP genotyping to accelerate the physical mapping of Rf2 locus in CMS-D8 cotton. The fertility of backcross population ((sterile line×restorer line)×maintainer line) comprising of 1623 individuals was investigated in the field. The fertile pool (100 plants with fertile phenotypes, F-pool) and the sterile pool (100 plants with sterile phenotypes, S-pool) were constructed for BSA resequencing. The selection of 24 single nucleotide polymorphisms (SNP) through high-throughput genotyping and the development insertion and deletion (InDel) markers were conducted to narrow down the candidate interval. The pentapeptide repeat (PPR) family genes and upregulated genes in restore line in the candidate interval were analysed by qRT-PCR. The fertility investigation results showed that fertile and sterile separation ratio was consistent with 1:1. BSA resequencing technology, high-throughput SNP genotyping, and InDel markers were used to identify Rf2 locus on candidate interval of 1.48 Mb on chromosome D05. Furthermore, it was quantified in this experiment that InDel markers co-segregated with Rf2 enhanced the selection of the restorer line. The qRT-PCR analysis revealed PPR family gene Gh_D05G3391 located in candidate interval had significantly lower expression than sterile and maintainer lines. In addition, utilization of anther RNA-Seq data of CMS-D8 identified that the expression level of Gh_D05G3374 encoding NB-ARC domain-containing disease resistance protein in restorer lines was significantly higher than that in sterile and maintainer lines. This study not only enabled us to precisely locate the restore gene Rf2 but also evaluated the utilization of InDel markers for marker assisted selection in the CMS-D8 Rf2 cotton breeding line. The results of this study provide an important foundation for further studies on the mapping and cloning of restorer genes.


CNS arousal systems: possible role in delirium.
Delirium is often considered a global and nonspecific alteration in cerebral function. However, the recent clinical evidence for heterogeneity within the syndrome of delirium suggests that different systems of the brain may be important in different kinds of delirium. Some forms of delirium, such as anticholinergic toxicity and hepatic encephalopathy, may be caused by drugs or toxins acting on specific brain neurochemical systems. The neurophysiological bases of the control of normal arousal and attention are still relatively poorly understood. However, the recent appreciation of the existence of neurotransmitter-specific projections from the hypothalamus and brain stem directly to the cerebral cortex has spurred new research. Some of these projections may be important in particular kinds of delirium; for instance, evidence from a number of different lines of research implicates GABA systems in the brain as being important in the delirium of hepatic encephalopathy. Cholinergic neurons in the basal forebrain and pons innervate the cerebral cortex. Both of these neurons, but particularly the pontine group, may be important in the delirium of anticholinergic toxicity. Thus, more research on the physiology of these systems in normal sleep and arousal, as well as in pathophysiological states, is indicated. Little is known about changes in these systems with aging. The well-known degeneration in cholinergic systems in Alzheimer's disease, and the sensitivity of individuals with Alzheimer's disease to anticholinergic toxicity, suggest a role of central cholinergic systems in anticholinergic delirium in demented patients. Further research into the involvement of the other systems in aging and delirium apparently would be fruitful.


Immunohistochemical evaluation of toxic epidermal necrolysis treated with human intravenous immunoglobulin.
Toxic epidermal necrolysis (TEN) is a severe drug reaction characterized by massive epidermal cell death. The authors of the current study and others have noted improved outcomes in TEN patients treated with human intravenous immunoglobulin (IVIG), purportedly due to its ability to inhibit the fas/fas-ligand (Fas-L) apoptotic pathway, but published case series evaluating TEN through the use of immunohistochemical antibody stains for Fas and Fas-L before and after IVIG treatment are lacking. The authors hypothesized that due to IVIG's ability to arrest the evolution of TEN, expression of Fas/Fas-L on keratinocytes would be decreased or absent following IVIG treatment. Ten patients diagnosed with TEN underwent biopsies of their lesions prior to and five days after treatment with IVIG. Seven post-treatment biopsies were of sufficient quality to undergo evaluation. All ten pretreatment biopsies had Fas and Fas-L expression by immunohistochemistry, while six out of seven (85.7%) post-treatment biopsies failed to demonstrate Fas or Fas-L expression. One of seven post-treatment biopsies stained positive for Fas and Fas-L. This reduced immunohistochemical expression of apoptotic markers may represent IVIG inhibition of the pathogenic mechanism of TEN. Alternatively reduced Fas and Fas-L may be a feature of reepithelialization in TEN, or characteristic of rapidly proliferating epidermis.


The renal degradation of myelin basic protein peptide 43-88 by two enzymes in different subcellular fractions.
Previous studies have demonstrated that the kidney is the major site for clearance and catabolism of a peptide (residues 43-88) of encephalitogenic or basic protein (BP) derived from central-nervous-system myelin. In the present investigation rat renal tissue was shown to be capable of degrading human BP peptide 43-88 over the pH range 4-11.5 with peaks of activity at pH5 and pH9. The enzymic activity at pH5 was localized mainly to the 5900 g pellet (crude mitochondrial fraction) and, on the basis of its sulphydryl features, was inferred to be cathepsin B. The enzyme activity at pH9 was greatly enriched in the 100 000 g pellet (microsomal brush-border fraction), and its sensitivity to inhibitors suggested that it was a metalloproteinase. The activity at alkaline pH in the 100 000 g pellet was stimulated 3-fold by non-ionic detergents and 20-fold by ATP and polyphosphates. Through a series of experiments the ATP stimulation of the alkaline proteinase activity was concluded to be the result of a reversal of inhibition imposed by the presence of another cationic protein, methylated bovine serum albumin. Inhibition by certain bivalent cations, the irregular effects of chelators and the effects of poly-L-lysine supported this conclusion. These studies indicate the availability of renal enzymes of different types and in different cellular compartments that are capable of degrading BP peptide 43-88. In particular, the relative amounts of bivalent cations, anions and charged proteins and peptides are likely to be major influences on the activity of the alkaline proteinase in vivo. The control of this degradation as well as the features of the smaller fragments of the peptide formed may determine biological and immune events subsequent to the release of this potentially autoantigenic material.