Factors Associated With Cannabis Use Among African American Nondaily Smokers.
Cannabis and tobacco dual use is a growing concern in the United States, especially among African Americans (AAs). Dual use increases nicotine dependence and poses negative health effects. Despite decreasing numbers of people who smoke daily, nondaily smokers (NDS) are increasing. Polytobacco use, including blunt use, is higher in AA NDS than AAs who smoke daily. This study examined factors associated with cannabis use among AA NDS. Adult AA NDS participated in a randomized controlled trial (n = 278) for smoking cessation. A subset of this sample (n = 262; mean age 48.2 years; 50% male) was analyzed to identify correlates of cannabis use. Logistic regression assessed the associations of demographic, smoking-related, and psychosocial variables with cannabis use. Participants smoked cigarettes on an average of 18 days of the last 30 and used 4.5 cigarettes on smoking days. Of the participants analyzed, 38% used cannabis, including blunts (ie, cigars hollowed out filled with cannabis) at baseline. Cannabis use was associated with polytobacco product use not including blunts (odds ratio [OR] 2.11, 95% confidence interval [CI] 1.18-3.77, P = 0.012), depressive symptoms (OR 1.22, 95% CI 1.05-1.42, P = 0.011), and younger age (OR 0.97, 95% CI 0.94-0.99, P = 0.004). Rates of cannabis and tobacco dual use in our sample exceed national rates. Dual use poses harmful health effects that exceed the risk of either substance alone. Findings will inform future work in tailoring treatments to vulnerable groups of people who use both tobacco and cannabis.


Picloram resistance in transgenic tobacco expressing an anti-picloram scFv antibody is due to reduced translocation.
Picloram resistance exhibited by transgenic tobacco (Nicotiana tabacum) plants expressing an anti-picloram single-chain variable fragment (scFv) antibody was investigated through the study of homozygous lines expressing the antibody. Dose-response bioassays, using foliar application of picloram, showed that these homozygous transgenic plants were resistant to at least 5 g of ai ha-1 picloram and grew normally to produce seed, whereas wild-type plants did not survive. Although these lines had improved resistance compared with those previously reported, significant improvements are still required to achieve field-level resistance. Uptake and translocation studies demonstrated that [14C]picloram translocation from treated leaves to the apical meristem was reduced in transgenic versus wild-type plants. The presence of [14C]picloram visualized by autoradiography and quantified by liquid scintillation spectrometry, demonstrated the distribution of more picloram in the treated leaf and less in the apical meristem of transgenic plants when compared to wild-type plants. No differences between transgenic and wild-type plants were found in the distribution of [14C]clopyralid, a herbicide with structural similarity to picloram as well as the same mechanism of action. No differences were found in the metabolism of [14C]picloram. Taken together, these results suggest that reduced translocation to the site of action is a major mechanism responsible for picloram resistance in tobacco plants expressing this anti-picloram antibody.


Finite-difference time-domain analysis of a complete transverse electromagnetic cell loaded with liquid biological media in culture dishes.
Transverse electromagnetic (TEM) cells can be used for exposing biological culture specimens to electromagnetic fields and observing possible anomalous effects. The uniformity of field exposure is critical to quantifying the biological response versus the electromagnetic dose. Standing waves and other electromagnetic field nonuniformities can cause nonuniform exposure. This paper reports the results of high-resolution three-dimensional finite-difference time-domain (FDTD) simulations of a complete TEM cell designed for operation at 837 MHz. Several different cases were studied in which the number of culture dishes, the depth of the culture liquid, and the orientation of the culture dishes were varied. Further, the effect of the culture-dish glass bottom thickness and the meniscus of the liquid medium were examined. The FDTD results show that there is a significant nonuniform field and specific absorption rate (SAR) distribution within the culture medium for each case examined. Hence, biological dose-response experiments using the TEM cells should account for the possibility of strong localized SAR peaking in the culture media to provide useful data in setting exposure standards for wireless communications.


Cortical excitability during prolonged antiepileptic drug treatment and drug withdrawal.
Previous reports characterized the effects of administration of single oral doses of antiepileptic drugs (AED) on cortical excitability. However, AED effects on cortical excitability, and their relationship to plasma blood levels, during chronic drug administration at therapeutic doses are not known. The objective of the study was to determine whether plasma blood levels during chronic administration at therapeutic doses would accurately predict changes in corticomotor excitability. We used transcranial magnetic stimulation (TMS) to measure cortical excitability during 5 weeks administration of carbamazepine (CBZ) and lamotrigine (LTG), and subsequent AED withdrawal in 20 healthy volunteers. Data were analyzed using ANOVA(RM) and regression analysis. Resting motor thresholds (r-MT) increased with increasing total and free CBZ and LTG levels during drug administration, but not drug withdrawal. After acute AED withdrawal, r-MT elevation persisted in most individuals with CBZ despite undetectable plasma levels, compared to a rapid normalization with LTG. In contrast, acute drug withdrawal resulted in a transient decrease in r-MT in 3/10 individuals with CBZ and 2/10 with LTG. Plasma levels provide information on motor cortical function during active treatment phases but not during AED withdrawal. The transient decrease in r-MT associated with acute AED withdrawal could represent a physiological substrate contributing to AED withdrawal seizures.


Weight-adjusted tinzaparin for the prevention of venous thromboembolism after bariatric surgery.
Essentials The optimal dose and duration of thromboprophylaxis after bariatric surgery are unclear. We evaluated the safety of weight-adjusted tinzaparin prophylaxis in 1212 patients. In-hospital rates of venous thromboembolism and major bleeding were 0.2% and 1.8% respectively. In a sub-set of patients, trough anti-Xa levels did not show excessive anticoagulant activity. Background Patients undergoing bariatric surgery are at moderate to high risk of venous thromboembolism (VTE). The optimal dose and duration of anticoagulant prophylaxis is uncertain. Objective To evaluate the safety of extended-duration weight-adjusted tinzaparin after bariatric surgery. Patients/methods We conducted a single-center retrospective cohort study of consecutive patients undergoing bariatric surgery who received weight-adjusted tinzaparin 4500-14 000 IU daily (75 IU kg-1 rounded to the nearest prefilled syringe) for 10 days after surgery (7-9 days post-hospital discharge). Primary safety outcomes were the frequency of VTE and major bleeding within 30 days of surgery in patients receiving at least one dose of tinzaparin. Results A total of 1279 patients undergoing bariatric surgery between July 2009 and December 2012 were reviewed, of whom 1212 received weight-adjusted tinzaparin. Safety outcomes were collected for 819 patients at 30 days, and for 1212 patients in-hospital only. The median age was 45.0 years, median weight was 130.0 kg and 98.8% of patients underwent gastric bypass or sleeve gastrectomy. In patients completing 30 days of follow-up, VTE occurred in 4/819 (0.5%) and major bleeding occurred in 13/819 patients (1.6%). In-hospital rates of VTE and major bleeding during surgical admission were 3/1212 (0.2%) and 22/1212 (1.8%), respectively. Conclusions Extended thromboprophylaxis with weight-adjusted tinzaparin appears to be a safe strategy after bariatric surgery, with low rates of postoperative VTE and major bleeding.


Surface tension transport of prey by feeding shorebirds: the capillary ratchet.
The variability of bird beak morphology reflects diverse foraging strategies. One such feeding mechanism in shorebirds involves surface tension-induced transport of prey in millimetric droplets: By repeatedly opening and closing its beak in a tweezering motion, the bird moves the drop from the tip of its beak to its mouth in a stepwise ratcheting fashion. We have analyzed the subtle physical mechanism responsible for drop transport and demonstrated experimentally that the beak geometry and the dynamics of tweezering may be tuned to optimize transport efficiency. We also highlight the critical dependence of the capillary ratchet on the beak's wetting properties, thus making clear the vulnerability of capillary feeders to surface pollutants.


Begin your partnership: the process of engagement.
Community Partnered-Participatory Research (CPPR) is based on and utilizes community engagement as its central method and principle. In this chapter, we explain the key differences between engaging the community vs merely involving the community. The chapter also reviews the plan-do-action cycle of work that is used in each stage of CPPR. We define five key values of CPPR: respect for diversity, openness, equality, redirected power (empowerment), and an asset-based approach. In addition, we present 12 operational principles, which guide work throughout every stage of all CPPR initiatives.


ADR in Journals: Are They Translated into Regulatory Frameworks?
An adverse drug reaction case report refers to a scientific publication that is written by a health care professional who suspects a casual relationship between a drug and an adverse drug reaction (ADR). ADR case reports help to identify potential risks associated with the use of drug. Most of the case reports do not mention about reporting the ADR to regulatory authorities. With this objective, the aim of this study was to analyze the number of Adverse Drug Reactions (ADR) published as case reports (PubMed indexed journals) from January 2018 to June 2019, and observe if they are translated in regulatory frameworks like Vigibase, and package inserts. 321 ADRs were obtained with the keywords "Adverse Drug Reaction". Out of those, 158 were independently extracted by two investigators, observed and categorized according to classes of the drugs, geographic location, severity, hospitalization, Completeness of ADR, whether reported to the regulatory authority (Vigibase), or listed in the package insert. Literature review articles were excluded. Out of the 158 ADRs, antibiotics accounted for 12.65%, CNS drugs and monoclonal antibodies11.39%, anticancer drugs 9.49%, CVS drugs 4.43%, anti-viral 3.79%, others 45.56%, respectively. According to geographic region, 26 ADRs published were from USA, Australia 4, Italy 3, India 17, Turkey 9, Singapore and UK 1, China 20, Denmark and Canada 2, Japan 10, France 9, Austria 1, Korea 5, South America 3, Switzerland 2, respectively. Depending upon the severity, causality assessment was done only for 45 ADRs, and not done for 113 ADRs. 41.13% patients (from 65 case reports) were hospitalized. Among the 158 ADRs, 14 ADRs were not found in Vigibase. 32 ADRs were not mentioned in the Drug package inserts. When categorized according to the completeness of case reports, weight accounted for1.89%, lab values and procedure for diagnosis, 96.8%, risk factors, 95.56%, prior exposure, 88.60%, Post ADR status, 60.12%, start-stop medication, route of administration, first dose, last dose, duration of illness accounted for 100%, respectively. Depending upon our observation, we have noticed that there is deficiency in reporting of suspected ADRs to regulatory authorities. Reporting can be included as mandatory criteria for ADR case reports. Also, there is an increased need to aware various healthcare workers for reporting ADR.


Calmodulin inhibits the protein kinase C-catalysed phosphorylation of an endogenous protein in A10 smooth-muscle cells.
The protein kinase C (PKC) activator phorbol 12,13-dibutyrate stimulated the phosphorylation of a 75 kDa protein (p75) in intact cultured A10 smooth-muscle cells and sonicated cell preparations; p75 was the only major substrate for endogenous PKC in sonicated A10 cells. The Ca(2+)-dependent phosphorylation of p75 in vitro was dramatically decreased in PKC-down-regulated A10 cells; however, p75 from identical sonicated cell preparations was still phosphorylated by an exogenous aortic PKC preparation. Calmodulin inhibited the phosphorylation of p75 by PKC, but not the phosphorylation of other PKC substrates (platelet P47 protein and histone). The addition of calmodulin after the phosphorylation reaction was started prevented further phosphorylation, but did not decrease the extent of phosphorylation of p75 that was reached before the addition of calmodulin. The inhibition of p75 phosphorylation was concentration-dependent, with IC50 values (concn. giving 50% inhibition) ranging from less than 0.5 to 10 micrograms of calmodulin/ml, and was Ca(2+)-dependent, requiring a free Ca2+ concentration of 10 microM or greater. These results suggest that the inhibition of the PKC-catalysed phosphorylation of p75 by calmodulin may be due to its interaction with the substrate, rather than a direct inhibitory effect on the enzyme, and that this inhibition could be regulated by intracellular Ca2+ concentration. Therefore, p75 may be a physiological link between the PKC and Ca2+/calmodulin pathways.


Antimicrobial resistance and distribution of sul genes and integron-associated intI genes among uropathogenic Escherichia coli in Queensland, Australia.
We studied 137 uropathogenic Escherichia coli (UPEC) isolates from hospitalized adult patients (Queensland, Australia) for their resistance to 17 antimicrobial agents using the calibrated dichotomous sensitivity method and the presence of class I, II and III integron-associated integrase (intI) genes, including functional class II intI2, as well as the presence of sul1, sul2 and sul3 genes, using PCR. Randomly amplified polymorphic DNA PCR, a high-resolution biochemical-fingerprinting method (PhP) and phylogenetic grouping were also used to identify the clonality of the sulphonamide-resistant isolates. One hundred and twenty (87.6 %) isolates were resistant to one or more of the tested antimicrobial drugs, with the highest resistance (70.1 %) observed against sulphafurazole (96 isolates). Of these, 84 (87.5 %) contained one or more sul alleles, with sul1 being the most common allele [occurring in 69 (72 %) isolates]. Only 38 of 69 (55.1 %) strains carrying the sul1 gene were positive for class I integrase. Our results indicate a high prevalence of sulphafurazole-resistant UPEC strains belonging to different clones among patients with urinary tract infection in Queensland, Australia. We also conclude that these strains carry predominantly a sul1 gene that is not commonly associated with the presence of class I integrase, indicating that it may be carried on either a bacterial chromosome or other genetic elements.


Associations between density and quality of health promotion programmes and built environment features across Jerusalem.
Health promotion programmes (HPPs) have the potential to influence individual health, depending on their quality and characteristics. Little is known about how they interact with built environment features and neighbourhood demographics in cities with substantial health disparities. Using the European Quality Instrument for Health Promotion (EQUIHP), we assessed the quality of HPPs, operating between 2016 and 2017, among adults aged 18-75 in Jerusalem. Areas were characterized by ethnicity and area socioeconomic level. Health information (body mass index, physical activity level) was obtained from the city profile survey. Geospatial information on the location and length of walking paths and bicycle lanes was obtained. Spearman correlations were used to assess associations among variables. Ninety-three HPPs operating in 349 locations in Jerusalem were identified. Programmes were unevenly distributed across urban planning areas (UPAs), with the highest density observed in the southwest, areas populated mainly by non-orthodox Jewish residents. However, the best performing HPPs based on EQUIHP score were in the north and east UPAs, inhabited primarily by Arab residents. At a neighbourhood level, characteristics of the built environment positively correlated with higher total EQUIHP scores: the ratio between walking lane length to the neighbourhood's population size (r = 0.413, P < 0.001) and length of bicycle lane per population (r = 0.309, P = 0.5). Median EQUIHP score negatively correlated with the number of programmes per neighbourhood size (m2) (r = -0.327, P = 0.006) and neighbourhood average socioeconomic status (SES; r = -0.266, P = 0.027). Our findings suggest that higher quality HPPs were preferentially located in areas of lower SES and served minority populations in Jerusalem.


Viability and ultrastructure of equine embryos following culture in a static or dynamic system.
The viability and ultrastructure of equine embryos were assessed following culture in a static or perifusion system. The percentage change in diameter was greater (P less than 0.025) for embryos in the static treatment (71%) than in the perifusion treatment (33%). Fluorescein diacetate (FD) scores, the percentage of fluorescing cells (FC) and fluorescent intensity (FI), also were greater (P less than 0.05 and P less than 0.01) following static culture than for embryos cultured in the perifusion system. Four of 9 control embryos resulted in pregnancies but no embryos cultured in either system prior to transfer resulted in pregnancies. Three of 9 embryos in the static treatment group and 1 of 9 embryos in the perifusion group resulted in non-ecogenic vesicles. Ultrastructural evaluation agreed with FD scores and morphological assessment. The continued development of static cultured embryos without the presence of an inner cell mass (ICM) was supported by ultrasound data.


Transplacental transmission and neonatal infection with swine influenza virus (Hsw1N1) in swine.
To study the question of chronicity or latency of swine influenza virus (Hsw1N1) infections in swine, newborn pigs were exposed to the virus in two experiments, and pregnant gilts were exposed in another experiment. Of five pigs exposed at 5 days of age, virus was isolated from throat swab samples of all (up to 10 postexposure days (PED) in one pig) and from a blood sample from one pig on PED 1 and 3. Virus was not isolated from urine, tissues, or explants of organs from pigs euthanatized PED 20 to 67, and disease was not evident. Of 11 pigs exposed within 2 hours of birth (before consuming colostrum), virus was shed for longer periods (for 10 and 11 days in four pigs) and severe respiratory tract pathologic changes developed. However, there was no evidence of chronic or latent infections. There was evidence of transplacental transmission of virus in one of ten pigs born to pregnant gilts that were exposed 10, 24, and 39 days before parturition, respectively.


Racial and ethnic differences in perceptions of campus climate related to sexual violence.
Objective: To assess perceptions of university institutional climate related to sexual violence and whether these differed by race/ethnicity. Participants: Matriculated undergraduates > age 18 (n = 1028). Methods: Students were invited via campus email to participate in an online survey. Results: Overall, only 20% agreed that the university is creating an environment in which unwanted sexual experiences seemed common or normal, but these findings differed by race. Black students were more likely than their white peers to feel the university is creating an environment in which unwanted sexual experiences seem common or normal (37.3% vs. 19.7%, p < .001) and creating an environment in which such instances were more likely to occur (33.3% vs. 13.4%, p < .001). Conclusions: Data suggest that while students generally perceive that the university is working to create a positive and safe climate, these perceptions vary by race. Further investigation is necessary to better understand the concerns of students of color.


Timosaponin AIII, a saponin isolated from Anemarrhena asphodeloides, ameliorates learning and memory deficits in mice.
Anemarrhena asphodeloides Bunge (AA, family Liliaceae), which primarily contains xantones, such as mangiferin, and steroidal saponins, such as timosaponin AIII and sarsasapogenin, has been used as an anti-pyretic, anti-inflammatory, anti-diabetic, anti-platelet aggregation, and anti-depressant agent in traditional Chinese medicine. In the present study, the memory-enhancing effects of these saponins were investigated in scopolamine-treated mice. Among saponins, timosaponin AIII (TA3) significantly reversed the scopolamine-induced deficits in a passive avoidance test and in the Morris water maze test. TA3 also increased hippocampal acetylcholine levels in scopolamine-treated mice and dose-dependently inhibited acetylcholinesterase (AChE) activity (IC(50) value, 35.4 microM). When TA3 (50 mg/kg) was orally administered to mice and its blood concentration was measured by liquid chromatography and tandem mass spectrometry, the C(max) of TA3 occurred 4-6 h after TA3 treatment. The memory-enhancing effect of TA3 was greater when it was administered 5 h before the acquisition trial than 1 h before. Scopolamine treatment in mice increased brain levels of TNF-alpha and IL-1beta expression. However, treatment with TA3 and scopolamine inhibited the increase of TNF-alpha and IL-1beta expression. These results suggest that scopolamine may cause learning and memory deficits that are further complicated by inflammation. TA3 also inhibited the activation of NF-kappaB signaling in BV-2 microglia and in SK-N-SH neuroblastoma cells induced with TNF-alpha or scopolamine. Nevertheless, TA3 may ameliorate memory deficits, mainly by inhibiting AChE.


Anatomic single- versus double-bundle ACL reconstruction: a meta-analysis.
To determine whether anatomic double-bundle anterior cruciate ligament (ACL) reconstruction compared to anatomic single-bundle ACL reconstruction more effectively restored antero-posterior (A-P) laxity, rotatory laxity and reduced frequency of graft rupture. Our hypothesis was that anatomic double-bundle ACL reconstruction results in superior rotational knee laxity and fewer graft ruptures due to its double-bundle tension pattern, compared with anatomic single-bundle ACL reconstruction. An electronic search was performed using the PubMed, EMBASE and Cochrane Library databases. All therapeutic trials written in English reporting knee kinematic outcomes and graft rupture rates of primary anatomic double- versus single-bundle ACL reconstruction were included. Only clinical studies of levels I-II evidence were included. Data regarding kinematic tests were extracted and included pivot-shift test, Lachman test, anterior drawer test, KT-1000 measurements, A-P laxity measures using navigation and total internal-external (IRER) laxity measured using navigation, as well as graft failure frequency. A total of 7,154 studies were identified of which 15 papers (8 randomized controlled trials and 7 prospective cohort studies, n = 970 patients) met the eligibility criteria. Anatomic ACL double-bundle reconstruction demonstrated less anterior laxity using KT-1000 arthrometer with a standard mean difference (SMD) = 0.36 (95% CI 0.214-0.513, p < 0.001) and less A-P laxity measured with navigation (SMD = 0.29 95% CI 0.01-0.565, p = 0.042). Anatomic double-bundle ACL reconstruction did not lead to significant improvements in pivot-shift test, Lachman test, anterior drawer test, total IRER or graft failure rates compared to anatomic single-bundle ACL reconstruction. Anatomic double-bundle ACL reconstruction is superior to anatomic single-bundle reconstruction in terms of restoration of knee kinematics, primarily A-P laxity. Whether these improvements of laxity result in long-term improvement of clinical meaningful outcomes remains uncertain. II.


Decoding the black box of health policy implementation: A case of regulating private healthcare establishments in southern India.
. Implementation of healthcare regulatory policies, especially in low- and middle-income countries where the private health sector is predominant, is challenging. Karnataka, a southern state in India, enacted the Karnataka Private Medical Establishments Act (KPMEA) with an aim to ensure quality of care in the private healthcare establishments. After more than a decade the implementation of KPMEA is suboptimal. . We used a case study design. The case was 'implementation of KPMEA'. The case study site was Bengaluru Urban district in Karnataka. Data from key informant interviews, focus group discussions held at the state, district and subdistrict levels and key policy documents, minutes of the meetings, data from the State Department of Health and Family Welfare, district level KPMEA data and litigations at the High Court of Karnataka were analysed using a framework. . The policy (KPMEA) content is inadequate and requires clarity in certain provisions of the Act. There was a lack of coordination between the implementing agencies. Workforce shortages were evident. Factors that impede the enforcement of the Act include poor knowledge and lack of competency of the officials on the content and the implementation mechanics of the policy, insufficient policy oversight from the state on the districts, corruption, political interference and lack of support from the local public, especially during raids on illegal establishments. . A regulatory policy such as KPMEA needs a clear, comprehensive content and directions for operationalization. However, improving the content of the policy is not easy as some aspects of the policy remain contentious with the private healthcare providers/ establishments. Addressing health governance issues at all levels is key to effective enforcement.


Tissue-engineered tendon nano-constructs for repair of chronic rotator cuff tears in large animal models.
Chronic rotator cuff tears (RCTs) are one of the most common injuries of shoulder pain. Despite the recent advances in surgical techniques and improved clinical outcomes of arthroscopically repaired rotator cuffs (RCs), complete functional recovery-without retear-of the RC tendon through tendon-to-bone interface (TBI) regeneration remains a key clinical goal to be achieved. Inspired by the highly organized nanostructured extracellular matrix in RC tendon tissue, we propose herein a tissue-engineered tendon nano-construct (TNC) for RC tendon regeneration. When compared with two currently used strategies (i.e., transosseous sutures and stem cell injections), our nano-construct facilitated more significant healing of all parts of the TBI (i.e., tendon, fibrocartilages, and bone) in both rabbit and pig RCT models owing to its enhancements in cell proliferation and differentiation, protein expression, and growth factor secretion. Overall, our findings demonstrate the high potential of this transplantable tendon nano-construct for clinical repair of chronic RCTs.


Managing Quartz Exposure in Apartment Building and Infrastructure Construction Work Tasks.
The present report describes exposure to respirable silica and dust in the construction industry, as well as means to manage them. The average exposure in studied work tasks (n = 148) amounted to 64% of the Finnish OEL value of 0.05 mg/m3. While 10% of exposure estimates exceeded the OEL, the 60% percentile was well below 10% of the OEL, as was the median exposure. In other words, exposure was low in more than half of the tasks. Work tasks where exposure was low included construction cleaning, work management, installation of concrete elements, rebar laying, driving work machines equipped with cabin air intake filtration, and landscaping, in addition to some road construction tasks. Excessive exposure (>OEL) was related to not using respiratory protection at all or not using it for long enough after the dusty activity ceased. Excessive exposures were found in sandblasting, dismantling facade elements, diamond drilling, drilling hollow-core slabs, drilling with a drilling rig, priming of explosives, tiling, use of cabinless earthmoving machines, and jackhammering, regardless of whether the hammering took place in an underpressurized compartment or not. Even in these tasks, it was possible to perform the work safely, following good dust prevention measures and, when necessary, using respiratory protection suitable for the job. Furthermore, in all tasks with generally low exposure, one could be significantly exposed through the general air or by making poor choices in terms of dust control.


Identification of a nuclear localization motif in the serine/arginine protein kinase PSRPK of physarum polycephalum.
Serine/arginine (SR) protein-specific kinases (SRPKs) are conserved in a wide range of organisms, from humans to yeast. Studies showed that SRPKs can regulate the nuclear import of SR proteins in cytoplasm, and regulate the sub-localization of SR proteins in the nucleus. But no nuclear localization signal (NLS) of SRPKs was found. We isolated an SRPK-like protein PSRPK (GenBank accession No. DQ140379) from Physarum polycephalum previously, and identified a NLS of PSRPK in this study. We carried out a thorough molecular dissection of the different domains of the PSRPK protein involved in its nuclear localization. By truncation of PSRPK protein, deletion of and single amino acid substitution in a putative NLS and transfection of mammalian cells, we observed the distribution of PSRPK fluorescent fusion protein in mammalian cells using confocal microscopy and found that the protein was mainly accumulated in the nucleus; this indicated that the motif contained a nuclear localization signal (NLS). Further investigation with truncated PSPRK peptides showed that the NLS (318PKKGDKYDKTD328) was localized in the alkaline Omega-loop of a helix-loop-helix motif (HLHM) of the C-terminal conserved domain. If the 318PKKGDK322 sequence was deleted from the loop or K320 was mutated to T320, the PSRPK fluorescent fusion protein could not enter and accumulate in the nucleus. This study demonstrated that the 318PKKGDKYDKTD328 peptides localized in the C-terminal conserved domain of PSRPK with the Omega-loop structure could play a crucial role in the NLS function of PSRPK.


Radiation Safety Considerations of Household Waste Disposal After Release of Patients Who Have Received [177Lu]Lu-PSMA-617.
Patients with metastatic prostate cancer are more likely than other groups to present for radiopharmaceutical therapy with urinary incontinence due to complications from prior local prostate cancer treatment. A consequence of urinary incontinence in patients receiving radiopharmaceutical therapy is the potential production of contaminated solid waste, which must be managed by the licensee and, at home, managed by and disposed of by the patient. Prolonging the patient stay in the treating facility after radiopharmaceutical therapy administration, until the first urinary void or potentially overnight, may moderately reduce the quantity of contaminated waste being managed by the patient at home. However, this approach does not fully mitigate the need for a patient waste-management strategy. In this brief communication, the relative radiation safety merits of contaminated waste disposal in the normal household waste stream in comparison to other waste management strategies are evaluated.


Short-acting P2Y12 blockade to reduce platelet dysfunction and coagulopathy during experimental extracorporeal circulation and hypothermia.
Extracorporeal circulation (ECC) and hypothermia are routinely used in cardiac surgery to maintain stable circulatory parameters and to increase the ischaemic tolerance of the patient. However, ECC and hypothermia cause platelet activation and dysfunction possibly followed by a devastating coagulopathy. Stimulation of the adenosinediphosphate (ADP) receptor P(2)Y(12) plays a pivotal role in platelet activation. This experimental study tested P(2)Y(12) receptor blockade as an approach to protect platelets during ECC. Human blood was treated with the short-acting P(2)Y(12) blocker cangrelor (1 µM, t(1/2)<5 min) or the P(2)Y(12) inhibitor 2-MeSAMP (100 µM) and circulated in an ex vivo ECC model at normothermia (37°C) and hypothermia (28°C). Before and after circulation, markers of platelet activation and of coagulation (thrombin-antithrombin complex generation) were analysed. During hypothermic ECC in pigs, the effect of reversible P(2)Y(12) blockade on platelet function was evaluated by cangrelor infusion (0.075 µg kg(-1) min(-1)). During ex vivo hypothermic ECC, P(2)Y(12) blockade inhibited platelet granule release (P<0.01), platelet-granulocyte binding (P<0.05), and platelet loss (P<0.001), whereas no effects on platelet-ECC binding, platelet CD42bα expression, glycoprotein IIb/IIIa activation, or thrombin-antithrombin complex generation were observed. During hypothermic ECC in pigs, cangrelor inhibited platelet-fibrinogen binding (P<0.05) and ADP-induced platelet aggregation (P<0.001). Platelet function was rapidly restored after termination of cangrelor infusion. P(2)Y(12) blockade by cangrelor prevents platelet activation during ECC and hypothermia. Owing to its short half-life, platelet inhibition can be well controlled, thus potentially reducing bleeding complications. This novel pharmacological strategy has the potential to reduce complications associated with ECC and hypothermia.


Radiation of the high-order plasmonic modes of large gold nanospheres excited by surface plasmon polaritons.
Large metallic nanoparticles with sizes comparable to the wavelength of light are expected to support high-order plasmon modes exhibiting resonances in the visible to near infrared spectral range. However, the radiation behavior of high-order plasmon modes, including scattering spectra and radiation patterns, remains unexplored. Here, we report on the first observation and characterization of the high-order plasmon modes excited in large gold nanospheres by using the surface plasmon polaritons generated on the surface of a thin gold film. The polarization-dependent scattering spectra were measured by inserting a polarization analyzer in the collection channel and the physical origins of the scattering peaks observed in the scattering spectra were clearly identified. More interestingly, the radiation of electric quadrupoles and octupoles was resolved in both frequency and spatial domains. In addition, the angular dependences of the radiation intensity for all plasmon modes were extracted by fitting the polarization-dependent scattering spectra with multiple Lorentz line shapes. A significant enhancement of the electric field was found in the gap plasmon modes and it was employed to generate hot-electron intraband luminescence. Our findings pave the way for exploiting the high-order plasmon modes of large metallic nanoparticles in the manipulation of light radiation and light-matter interaction.


Diabetes risk score in the United Arab Emirates: a screening tool for the early detection of type 2 diabetes mellitus.
The objective of this study was to develop a simple non-invasive risk score, specific to the United Arab Emirates (UAE) citizens, to identify individuals at increased risk of having undiagnosed type 2 diabetes mellitus. A retrospective analysis of the UAE National Diabetes and Lifestyle data was conducted. The data included demographic and anthropometric measurements, and fasting blood glucose. Univariate analyses were used to identify the risk factors for diabetes. The risk score was developed for UAE citizens using a stepwise forward regression model. A total of 872 UAE citizens were studied. The overall prevalence of diabetes in the UAE adult citizens in the Northern Emirates was 25.1%. The significant risk factors identified for diabetes were age (≥35 years), a family history of diabetes mellitus, hypertension, body mass index ≥30.0 and waist-to-hip ratio ≥0.90 for males and ≥0.85 for females. The performance of the model was moderate in terms of sensitivity (75.4%, 95% CI 68.3 to 81.7) and specificity (70%, 95% CI 65.8 to 73.9). The area under the receiver-operator characteristic curve was 0.82 (95% CI 0.78 to 0.86). A simple, non-invasive risk score model was developed to help to identify those at high risk of having diabetes among UAE citizens. This score could contribute to the efficient and less expensive earlier detection of diabetes in this high-risk population.


Measurements of personal UV exposure on different parts of the body during various activities.
Within this study, the erythemal ultraviolet (UV) exposure received by different parts of the body during four different activities is determined. Optoelectronic devices were used to measure the erythemal UV exposure at 10 different positions of the body. The measuring devices were fixed on the forehead, on the shoulders, on the arms, on the chest, on the thighs and on the lower legs. The measurements were performed during the following activities of the test persons: walking, sitting, lying and sitting up. The measurements were performed on four clear sky days in the early afternoon at 1 s interval. One measurement sequence was taking 30-40 min. For the analysis of the measured UV exposures, the ambient UV is taken as a reference to remove the atmospheric fluctuations on the measured UV exposure. The strong dependence of the UV exposure on the activity and on the orientation of the test person is shown. Most of the body parts receive the highest exposure, when the test subject is sitting up or lying. The shoulders are most at risk when the test person is walking, whereas during the activities sitting up and lying the legs are most at risk.


Lymphovascular invasion in micropapillary urothelial carcinoma: a study of 22 cases.
Micropapillary urothelial carcinoma (MPUC) is a known aggressive variant of urothelial carcinoma. However, the reasons for its aggressiveness remain unclear. To investigate the frequency of lymphovascular invasion in 22 cases of MPUC. Consecutive tissue sections were stained with D2-40 and CD34 to highlight lymphovascular channels associated with MPUC. Spaces containing tumor cells were scored as positive for lymphovascular invasion if the staining pattern on immunohistochemistry was distinct and circumferential. Of 22 cases, 21 (95%) had lymphovascular invasion on immunohistochemical staining, with 91% lymphatic invasion and 4% vascular invasion. Interestingly, 8 cases were originally signed out as negative for lymphovascular invasion on the basis of hematoxylin-eosin-stained sections; of these, 7 (88%) had focal lymphovascular invasion evident on immunohistochemical staining. Our results confirm that micropapillary lacunae are not lymphovascular channels. However, nearly all MPUC tumors (95% in this series) have evidence of lymphovascular invasion by immunohistochemical analysis. Our data support the use of micropapillary features as a morphologic marker for lymphovascular invasion and MPUC as an adverse histologic type of urothelial carcinoma.


Thermodynamic signature of substrates and substrate analogs binding to human blood group B galactosyltransferase from isothermal titration calorimetry experiments.
It has been observed earlier that human blood group B galactosyltransferase (GTB) hydrolyzes its donor substrate UDP-Galactose (UDP-Gal) in the absence of acceptor substrate, and that this reaction is promoted by the presence of an acceptor substrate analog, α-L-Fuc-(1,2)-β-D-3-deoxy-Gal-O-octyl (3DD). This acceleration of enzymatic hydrolysis of UDP-Gal was traced back to an increased affinity of GTB toward the donor substrate in the presence of 3DD. Herein, we present new thermodynamic data from isothermal titration calorimetry (ITC) on the binding of donor and acceptor substrates and analogs to GTB. ITC data are supplemented by surface plasmon resonance and STD-NMR titration experiments. These new data validate mutual allosteric control of binding of donor and acceptor substrates to GTB. It is of note that ITC experiments reveal significant differences in enthalpic and entropic contributions to binding of the natural donor substrate UDP-Gal, when compared with its analog UDP-Glucose (UDP-Glc). This may reflect different degrees of ordering of an internal loop (amino acids 176-194) and the C-terminus (amino acids 346-354), which close the binding pocket on binding of UDP-Gal or UDP-Glc. As both ligands have rather similar dissociation constants KD and almost identical modes of binding this finding is unexpected. Another surprising finding is that an acceptor analog, α-L-Fuc-(1,2)-β-D-3-amino-3-deoxy-Gal-O-octyl (3AD) as well as the constituent monosaccharide β-D-3-amino-3-deoxy-Gal-O-octyl (3AM) effectively inhibit enzymatic hydrolysis of UDP-Gal. This is unexpected, too, because in analogy to the effects of 3DD one would have predicted acceleration of enzymatic hydrolysis of UDP-Gal. It is difficult to explain these observations based on structural data alone. Therefore, our results highlight that there is an urgent need of experimental studies into the dynamic properties of GTB.


Genetic and biochemical studies of hepatic carcinoma in the Egyptian population.
Hepatocellular carcinoma (HCC), a deadly malignancy of the liver, is considered the third leading reason behind cancer deaths. It is more frequent in men than in women of ages above 50. Liver disease, leading to liver cirrhosis (LC), is mostly caused by alcoholism abuse, reaction diseases of the liver, or viral hepatitis B or C infection. Interleukin-6 (IL-6) is considered an effective pro-inflammatory cytokine, which plays a crucial role in the host defense mechanism. Its level is higher in HCC patients than in LC cases, indicating that tumor cells increase the production of cytokines. The X-ray repair cross-complementing group 1 (XRCC1) gene is a major DNA repair gene. It acts as a scaffold of various activities that are concerned in the repairing method by interacting with components of base excision repair. This study aims to measure the serum concentrations of IL6 and C-reactive protein (CRP) and investigate whether XRCC1 Arg194Trp and Arg399Gln polymorphisms are related to HCC disease. Whole-blood DNA was extracted from 123 HCC patients and 123 healthy volunteers. Tetra-primer amplification refractory mutation system was performed in the detection of XRCC1 Arg399Gln and Arg194Trp polymorphisms. Serum concentration levels of IL-6 and CRP are significantly higher in patients with HCC than in control subjects. The allelic and genotype frequency distributions of XRCC1 (Arg399Gln and Arg194Trp) are significantly increased in HCC cases compared to healthy volunteers. Arg/Gln, Arg/Trp, Gln/Gln, and Trp/Trp genotypes are associated with higher risk HCC than the Arg/Arg genotype.


PU.1 regulates both cytokine-dependent proliferation and differentiation of granulocyte/macrophage progenitors.
PU.1 is a unique regulatory protein required for the generation of both the innate and the adaptive immune system. It functions exclusively in a cell-intrinsic manner to control the development of granulocytes, macrophages, and B and T lymphocytes. We demonstrate that mutation of the PU.1 gene causes a severe reduction in myeloid (granulocyte/macrophage) progenitors. PU.1 -/- myeloid progenitors can proliferate in vitro in response to the multilineage cytokines interleukin-3 (IL-3), IL-6 and stem cell factor but are unresponsive to the myeloid-specific cytokines granulocyte-macrophage colony-stimulating factor (GM-CSF), G-CSF and M-CSF. The failure of PU.1 -/- progenitors to respond to G-CSF is bypassed by transient signaling with IL-3. In the presence of IL-3 and G-CSF, PU.1 -/- progenitors can differentiate into granulocytic precursors containing myeloperoxidase-positive granules. Thus PU.1 is not essential for specification of granulocytic precursors, but is required for their further differentiation. The failure of PU.1 -/- progenitors to respond to M-CSF is due to lack of c-fms gene transcription. Transduction of c-fms into PU.1 -/- myeloid progenitors bypasses the block to M-CSF-dependent proliferation but does not induce detectable macrophage differentiation. Therefore, PU. 1 appears to be essential for specification of monocytic precursors. Importantly, retroviral transduction of PU.1 into mutant progenitors restores responsiveness to myeloid-specific cytokines and development of mature granulocytes and macrophages. Thus PU.1 controls myelopoiesis by regulating both proliferation and differentiation pathways.


Combined carotid stenting and urgent coronary artery surgery in unstable angina patients with severe carotid stenosis.
Staged or combined carotid endarterectomy (CEA) offers the potential benefit of decreased neurological morbidity during and after cardiac surgery; however, the strategy for treating unstable high-risk patients, who need urgent coronary artery surgery, remains unresolved. We report in-hospital and 30-day outcomes of 23 consecutive patients admitted with unstable angina, who underwent carotid angioplasty and stenting (CAS) immediately prior to urgent coronary artery surgery, from October 2007 to October 2008. Aspirin and unfractioned heparin were administrated during carotid stenting and clopidogrel was only started after cardiac surgery. All patients remained event-free during and immediately after the carotid stenting procedure. One patient died due to sepsis 22 days after cardiac surgery. There was neither stroke nor myocardial infarction at follow-up. No patient needed a cardiac or carotid re-intervention. This new approach (combined carotid stenting and coronary artery surgery) provides a less radical intervention, can be performed with a low periprocedural complication rate and may become a valuable alternative in the treatment of high-risk patients with combined carotid and cardiac disease.


Expression of Hey marks a subset of enteroendocrine cells in the Drosophila embryonic and larval midgut.
Hey is a conserved transcription factor of the bHLH-Orange family that participates in the response to Notch signaling in certain tissues. Whereas three Hey paralogues exist in mammalian genomes, Drosophila possesses a single Hey gene. Fly Hey is expressed in the subset of newborn neurons that receive a Notch signal to differentiate them from their sibling cells after the asymmetric division of precursors called ganglion-mother-cells. We used a polyclonal anti-Hey serum and a GFP-tagged transgenic duplication of the Hey locus to examine its expression in tissues outside the nervous system in embryos and larvae. We detected robust Hey expression in the embryonic midgut primordium at the time of birth of enteroendocrine cells, identified by expression of Prospero. Approximately half of the Pros-positive cells were also Hey positive at mid-embryogenesis. By the end of embryogenesis, most enteroendocrine cells had downregulated Hey expression, although it was still detectable at low levels after hatching. Low levels of Hey were also detected in subsets of the epithelial enterocytes at different times. Embryo enteroendocrine Hey expression was found to be Notch dependent. In late third-instar larvae, when few new enteroendocrine cells are born, novel Hey expression was detected in one cell of each sibling pair. In conclusion, Hey is strongly expressed in one of each pair of newly-born enteroendocrine cells. This is consistent with a hypothesis that embryonic enteroendocrine cells are born by an asymmetric division of a precursor, where Notch/Hey probably distinguish between the subtypes of these cells upon their differentiation.


Evaluation of cortisol precursors for the diagnosis of pituitary-dependent hypercortisolism in dogs.
The serum concentrations of cortisol, 17alpha-hydroxypregnenolone, 17alpha-hydroxyprogesterone, 21-deoxycortisol and 11-deoxycortisol were measured in 19 healthy dogs, 15 dogs with pituitary-dependent hypercortisolism (pdh) and eight dogs with other diseases before and one hour after an injection of synthetic adrenocorticotrophic hormone (acth). At both times the dogs with pdh had significantly higher concentrations of cortisol, 17alpha-hydroxypregnenolone, 17alpha-hydroxyprogesterone and 21-deoxycortisol than the healthy dogs. Basal 11-deoxycortisol concentrations were also significantly higher in dogs with pdh compared with healthy dogs. When compared with the dogs with other diseases, the dogs with pdh had significantly higher basal and post-acth cortisol and basal 21-deoxycortisol, and significantly lower post-acth 11-deoxycortisol concentrations. The dogs with other diseases had significantly higher post-acth cortisol, 17alpha-hydroxyprogesterone and 11-deoxycortisol concentrations than the healthy dogs. In general, the post-acth concentrations of 17alpha-hydroxypregnenolone, 17alpha-hydroxyprogesterone, 11-deoxycortisol and 21-deoxycortisol were more variable than the post-acth concentrations of cortisol, resulting in large overlaps of the concentrations of these hormones between the three groups. A two-graph receiver operating characteristic (ROC) analysis was used to maximise the sensitivity and specificity of each hormone for diagnosing hypercortisolism; it showed that the post-acth concentration of cortisol had the highest sensitivity and specificity. The overlaps between the healthy dogs, the dogs with pdh and the dogs with other diseases suggested that the individual precursor hormones would not be useful as a screening test for hypercortisolism.


Acute sterile endophthalmitis following intravitreal bevacizumab: case series.
Since the ophthalmological community adopted the use of intravitreal bevacizumab as an accepted off-label treatment for neovascular diseases, the amount of knowledge regarding its effects and properties has been increasing continually. In the last few years, there have been an increasing number of reports about sterile intraocular inflammation and intraocular pressure elevations after intravitreal bevacizumab. In the following case series, we describe the clinical presentation and outcomes of ten consecutive cases of patients developing mild-to-severe sterile intraocular inflammation after intravitreal bevacizumab and their management. This report presents a retrospective case series. We reviewed the medical records of ten consecutive patients from a group of 46, in whom repackaged bevacizumab in individual aliquots from two vials from the same batch were used. All surgical procedures were performed using standard sterile techniques in the operating room. At each follow-up visit, patients underwent a complete ophthalmological examination including visual acuity assessment, intraocular pressure, biomicroscopy, and posterior fundus examination. Ten patients presented sterile endophthalmitis with an onset time of 3.5±1.95 days. The clinical characteristics were mild pain, slight visual loss, conjunctival hyperemia, and various degrees of intraocular inflammation with microhypopyon. All cultures were negative. All patients were managed with topical steroids and antibiotics, except two, in whom, due to severe vitreous cells, intravitreal antibiotics were used. Three patients showed a transient elevation of intraocular pressure. Only 50% of the patients regained a visual acuity equal or better to the baseline visual acuity on file. The increasing number of intravitreal injections of bevacizumab applied every day, due to its widespread acceptance, might be one reason why the number of cases of sterile endophthalmitis is rising. Fast recognition and accurate differential diagnosis is important to avoid unnecessary treatments and long-term complications. The low incidence of this event should not preclude the use of intravitreal injections in eyes that could benefit greatly from this therapy.


Protein actors sustaining arbuscular mycorrhizal symbiosis: underground artists break the silence.
The roots of most land plants can enter a relationship with soil-borne fungi belonging to the phylum Glomeromycota. This symbiosis with arbuscular mycorrhizal (AM) fungi belongs to the so-called biotrophic interactions, involving the intracellular accommodation of a microorganism by a living plant cell without causing the death of the host. Although profiling technologies have generated an increasing depository of plant and fungal proteins eligible for sustaining AM accommodation and functioning, a bottleneck exists for their functional analysis as these experiments are difficult to carry out with mycorrhiza. Nonetheless, the expansion of gene-to-phenotype reverse genetic tools, including RNA interference and transposon silencing, have recently succeeded in elucidating some of the plant-related protein candidates. Likewise, despite the ongoing absence of transformation tools for AM fungi, host-induced gene silencing has allowed knockdown of fungal gene expression in planta for the first time, thus unlocking a technological limitation in deciphering the functional pertinence of glomeromycotan proteins during mycorrhizal establishment. This review is thus intended to draw a picture of our current knowledge about the plant and fungal protein actors that have been demonstrated to be functionally implicated in sustaining AM symbiosis mostly on the basis of silencing approaches.


Psychiatric symptoms in adolescents with Internet use: comparison without Internet use.
The purpose of the present study was to investigate the psychiatric symptoms of adolescents with and without Internet use. A total of 2,080 students (1,105 male, 975 female) were recruited for the study. We administered the Symptom Checklist-90-Revision and Questions Related to Internet Use. Students in 10th through 12th grades were recruited from 14 senior high schools in Sivas, Turkey. Data were collected from October to May 2008. Data were analyzed after excluding the participants who provided incomplete data. It was found that Internet use in adolescents was associated with more severe psychiatric symptoms. Attention should be devoted to adolescents with Internet use for the design of preventive strategies.


Dissection of the asynchronous transport of intestinal microvillar hydrolases to the cell surface.
Novel subcellular fractionation procedures and pulse-chase techniques were used to study the intracellular transport of the microvillar membrane hydrolases sucrase-isomaltase and dipeptidylpeptidase IV in the differentiated colon adenocarcinoma cell line Caco-2. The overall rate of transport to the cell surface was two fold faster for dipeptidylpeptidase IV than for sucrase-isomaltase, while no significant differences were observed in transport rates from the site of complex glycosylation to the brush border. The delayed arrival of sucrase-isomaltase in the compartment where complex glycosylation occurs was only in part due to exit from the endoplasmic reticulum. A major slow-down could be ascribed to maturation in and transit of this enzyme through the Golgi apparatus. These results suggest that the observed asynchronism is due to more than one rate-limiting step along the rough endoplasmic reticulum to trans-Golgi pathway.


Distinct mast cell subpopulations within and around lymphatic vessels regulate lymph flow and progression of inflammatory-erosive arthritis in TNF-transgenic mice.
Inflammatory-erosive arthritis is exacerbated by dysfunction of joint-draining popliteal lymphatic vessels (PLVs). Synovial mast cells are known to be pro-inflammatory in rheumatoid arthritis (RA). In other settings they have anti-inflammatory and tissue reparative effects. Herein, we elucidate the role of mast cells on PLV function and inflammatory-erosive arthritis in tumor necrosis factor transgenic (TNF-tg) mice that exhibit defects in PLVs commensurate with disease progression. Whole mount immunofluorescent microscopy, toluidine blue stained histology, scanning electron microscopy, and in silico bioinformatics were performed to phenotype and quantify PLV mast cells. Ankle bone volumes were assessed by μCT, while corresponding histology quantified synovitis and osteoclasts. Near-infrared indocyanine green imaging measured lymphatic clearance as an outcome of PLV draining function. Effects of genetic MC depletion were assessed via comparison of 4.5-month-old WT, TNF-tg, MC deficient KitW-sh/W-sh (cKit-/-), and TNF-tg x cKit-/- mice. Pharmacological inhibition of mast cells was assessed by treating TNF-tg mice with placebo or cromolyn sodium (3.15mg/kg/day) for 3-weeks. PLVs are surrounded by MCT+/MCPT1+/MCPT4+ mast cells whose numbers are increased 2.8-fold in TNF-tg mice. The percentage of peri-vascular degranulating mast cells was inversely correlated with ICG clearance. A population of MCT+/MCPT1-/MCPT4- mast cells were embedded within the PLV structure. In silico single-cell RNA-seq (scRNAseq) analyses identified a population of PLV-associated mast cells (marker genes: Mcpt4, Cma1, Cpa3, Tpsb2, Kit, Fcer1a & Gata2) with enhanced TGFβ-related signaling that are phenotypically distinct from known MC subsets in the Mouse Cell Atlas. cKit-/- mice have greater lymphatic defects than TNF-tg mice with exacerbation of lymphatic dysfunction and inflammatory-erosive arthritis in TNF-tg x cKit-/- vs. TNF-Tg mice. Cromolyn sodium therapy stabilized PLV mast cells, increased TNF-induced bone loss, synovitis, and osteoclasts, and decreased ICG clearance. Mast cells are required for normal lymphatic function. Genetic ablation and pharmacological inhibition of mast cells exacerbates TNF-induced inflammatory-erosive arthritis with decreased lymphatic clearance. Together, these findings support an inflammatory role of activated/degranulated peri-PLV mast cells during arthritic progression, and a homeostatic role of intra-PLV mast cells, in which loss of the latter dominantly exacerbates arthritis secondary to defects in joint-draining lymphatics, warranting investigation into specific cellular mechanisms.


Use of the laryngeal mask airway in managing a patient with a large anterior mediastinal mass: a case report.
This is a case presentation of a patient with a large anterior mediastinal mass in whom the airway was maintained with a laryngeal mask airway. The patient was otherwise managed in the "classic" fashion with spontaneous respiration via a light general anesthetic. In reviewing many of the potential pitfalls of anesthesia in this class of patients, the laryngeal mask airway was found to be both a valuable and viable option for airway management.


Direct in situ reverse transcriptase-polymerase chain reaction.
In situ hybridization has been used for localization of specific nucleic acid sequences at the cellular level despite providing relatively low-detection sensitivity. In situ reverse transcriptase-polymerase chain reactions (RT-PCR) enhance sensitivity and thus enable localization of low-abundance mRNA in a cell. However, the available methods are fraught with problems of nonspecific amplifications as a result of mispriming and/or amplification from partially digested residual genomic DNA in tissue. Herein, we demonstrate that nonspecific background amplification can be eliminated by pretreatment of samples with restriction enzymes before DNase I digestion. Primers tagged with a far-red shifted fluorescent dye such as Cy5 in PCR reactions allow identification of target mRNA by fluorescence microscopy. These novel modifications lead to increased specificity and rapid in situ detection of cellular mRNA and thus may be used for pathological diagnosis.


Eliminating disparities in hypertension care for Hispanics and blacks using a heart failure disease management program.
This study assessed if patients enrolled in a heart failure disease management program (HFDMP) reach the JNC VII target goals for blood pressure (BP) control, eliminate disparities in hypertension control by race/ ethnicity and the impact BP control has on survival. Patients (N = 898) with an ejection fraction <40% were enrolled into two HFDMPs and screened for hypertension, defined as BP > 130/80. Mean baseline systolic BP (SBP) 132 ± 25.5 mm Hg and diastolic BP (DBP) 79 ± 16.8 mm Hg. Final mean SBP decreased to 129.6 mm Hg, DBP 77.6 mm Hg. Whites had the highest rate of achieving BP goals. Mortality reduction was associated with minority race, history of hypertension, increase ejection fraction and statin use. HFDMPs are an effective way to reduce BP in hypertensive patients. Disparities by race and ethnicity were not seen after adjustment for disease modifiers. There was no mortality difference in those who reached BP goal.


Peroxidase-like activity of ferruginous bodies isolated by exploiting their magnetic property.
Ferruginous bodies (FB) are polymorphic structures whose formation is macrophage dependent, and are composed of a core, which may consist of an asbestos fiber coated with proteins, among which ferritin is the main component. Within ferritin, the ferric and ferrous ions are coordinated as ferrihydrite, which is the main iron (Fe) storage compound. However, when ferritin accumulates in some tissues following Fe overload it also contains magnetite along with ferrihydrite, which endows it with magnetic properties. Recently studies showed that magnetite exerts peroxidase-like activity, and since ferruginous bodies display magnetic properties, it was postulated that these particular structures may also contain magnetite within the ferritin coating, and thus may also exert peroxidase-like activity. Histochemical analysis for peroxidase of isolated FB smears demonstrated positive staining. Samples isolated from 4 different autopsy lung fragments were also able to oxidize 3,3',5,5'-tetramethyl-benzidine to a blue colored compound that absorbs at 655 nm. This activity was (1) azide and heat insensitive with optimal pH from 5 to 6, and (2) highly variable, changing more than 25-fold from one sample to another. These findings, together with evidence that the peroxidase-like activity of ferruginous bodies has a hydrogen peroxide and substrate requirement different from that of human myeloperoxidase, can exclude that this enzyme gives a significant contribution to the formation of FB. Standard Fe-rich asbestos fibers also express a peroxidase-like activity, but this appears negligible compared to that of ferruginous bodies. Strong acidification of standard Fe-containing asbestos fibers or magnetically isolated ferruginous bodies liberates a high amount of peroxidase-like activity, which is probably accounted for by the release of Fe ions. Further, FB also damage mesothelial cells in vitro. Data suggest that FB exert peroxidase-like activity and cytotoxic activity against mesothelial cells, and hence may be an important factor in pathogenesis of asbestos-related diseases.


Cardiovascular adjustments to acute hypoxemia superimposed on chronic hypoxemia in lambs.
Cardiovascular responses to acute hypoxemia are in part mediated through adrenergic and chemoreceptor stimulation. In chronic hypoxemia the response to these stimuli may be blunted. Therefore, we determined whether the cardiovascular responses to acute hypoxemia superimposed on 3-4 wk of chronic hypoxemia were blunted in lambs with an experimental cardiac right-to-left shunt (combination of atrial septal defect and variable pulmonary stenosis). Cardiovascular variables and regional blood flows were determined during chronic hypoxemia and after acutely reducing the arterial oxygen saturation by increasing the cardiac right-to-left shunt. Arterial oxygen saturation decreased (65 +/- 7 to 40 +/- 7%, P < 0.001) and systemic blood flow increased (164 +/- 63 to 233 +/- 100 ml.min-1.kg-1, P < 0.01), maintaining systemic oxygen supply and oxygen uptake. Blood flow to the myocardium (P < 0.01), the adrenals (P < 0.05), and the brain (0.05 < P < 0.10) increased, and oxygen supply to these organs was maintained. Conversely, blood flow to the kidneys and the gastrointestinal tract was unaltered, so that oxygen supply to these organs was decreased. The responses to acute hypoxemia in chronically hypoxemic lambs were similar to those previously reported in normoxemic lambs. We conclude that the cardiovascular responses to acute hypoxemia in chronically hypoxemic lambs are not blunted.


Superfluorescent squaraine with efficient two-photon absorption and high photostability.
The synthesis, linear photophysical, two-photon absorption (2PA), femtosecond transient absorption, and superfluorescence properties of a new symmetrical squaraine derivative (1) are reported. Steady-state linear spectral and photochemical properties, fluorescence lifetimes, and excitation anisotropy of 1 were investigated in various organic solvents. High fluorescence quantum yields (≈0.7) and very high photostability (photodecomposition quantum yields ≈10(-6)-10(-8)) were observed. An open-aperture Z-scan method was used to obtain 2PA spectra of 1 over a broad spectral range (maximum 2PA cross section ≈1000 GM). Excited-state absorption (ESA) and gain was observed by femtosecond transient absorption spectroscopy, in which both reached a maximum at approximately 500 fs. Squaraine 1 exhibits efficient superfluorescence. The quantum chemical study of 1 revealed the simulated vibronic nature of the 1PA and 2PA spectra were in good agreement with experimental data; this may provide the ability to predict potential advanced photonic materials.


Rapid molecular mass and structural determination of plant cell wall-derived oligosaccharides using off-line high-performance anion-exchange chromatography/mass spectrometry.
A method has been developed for the rapid molecular mass determination and structural elucidation of mixtures of oligosaccharides derived from plant cell walls. The oligosaccharides were fractionated using gel permeation chromatography and 'analytical' high-performance anion-exchange chromatography (HPAEC), neutralized, dried and the mixtures of eluent salt and oligosaccharides were per-O-acetylated directly. The derivatized oligosaccharides were isolated by dissolution in dichloromethane and the salts were removed by aqueous partitioning. The per-O-acetylated oligosaccharides were analysed using electrospray (ES) and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MS). Exploiting the fact that acid-catalysed per-O-acetylation of oligosaccharides can be achieved even under the extremely salty conditions that are found in post-column neutralized HPAEC fractions, and combining this derivatization step with off-line ESMS, allow rapid screening for molecular mass and thus yield information on the composition of the various oligosaccharides in these complex mixtures. Subsequent per-O-methylation of the per-O-acetylated, salt-free fractions and collision-induced dissociation tandem mass spectrometric analysis was used for additional sequence and branching determination of the oligosaccharides.


Repeat Bone Mineral Density Screening Measurement and Fracture Prediction in Older Men: A Prospective Cohort Study.
Whether repeated bone mineral density (BMD) screening improves fracture prediction in men is uncertain. We evaluated whether a second BMD 7 years after the initial BMD improves fracture prediction in older men. Among 3651 community-dwelling men (mean age 79.1 years) with total hip BMD at baseline and Year 7 (Y7), self-reported fractures after Y7 were confirmed by radiographic reports. Fracture prediction assessed using Cox proportional hazards regression and logistic regression with receiver operating characteristic curves for models based on initial BMD, BMD change, and the combination of initial BMD and BMD change (combination model). During an average follow-up of 8.2 years after Y7, 793 men experienced ≥ 1 clinical fractures, including 426 men with major osteoporotic fractures (MOF) and 193 men with hip fractures. Both initial BMD and BMD change were associated with risk of fracture outcomes independent of each other, but the association was stronger for initial BMD. For example, the multivariable hazard ratio of MOF in the combination model per 1 SD decrement in BMD was 1.76 (95% CI 1.57-1.98) for initial BMD and 1.19 (95% CI 1.08-1.32) for BMD change. Discrimination of fracture outcomes with initial BMD models was somewhat better than with BMD change models and similar to combination models (AUC value for MOF 0.68 [95% CI 0.66-0.71] for initial BMD model, 0.63 [95% CI 0.61-0.66] for BMD change model, and 0.69 [95% CI 0.66-0.71] for combination model). Repeating BMD after 7 years did not meaningfully improve fracture prediction at the population level in community-dwelling older men.


Premonitory sensory phenomenon in Tourette's syndrome.
We administered a questionnaire designed to probe for premonitory sensations associated with motor tics to 50 patients with Tourette's syndrome (TS). Premonitory sensations were reported by 46 (92%) patients, and the most common sensation was an urge to move and an impulse to tic ("had to do it"). Intensification of premonitory sensations, if prevented from performing a motor tic, was reported also in 37 patients (74%), 36 patients (72%) reported relief of premonitory sensations after performing the tic, and 27 of 40 (68%) described a motor tic as a voluntary motor response to an involuntary sensation, rather than a completely involuntary movement. The "just right" sensation correlated with the presence of co-morbid obsessive-compulsive disorder. We conclude that premonitory sensations are an important aspect of motor tics and some patients perceive motor tics as a voluntary movement in response to an involuntary sensation.


Continuous monitoring of prostate position using stereoscopic and monoscopic kV image guidance.
To demonstrate continuous kV x-ray monitoring of prostate motion using both stereoscopic and monoscopic localizations, assess the spatial accuracy of these techniques, and evaluate the dose delivered from the added image guidance. The authors implemented both stereoscopic and monoscopic fiducial localizations using a room-mounted dual oblique x-ray system. Recently developed monoscopic 3D position estimation techniques potentially overcome the issue of treatment head interference with stereoscopic imaging at certain gantry angles. To demonstrate continuous position monitoring, a gold fiducial marker was placed in an anthropomorphic phantom and placed on the Linac couch. The couch was used as a programmable translation stage. The couch was programmed with a series of patient prostate motion trajectories exemplifying five distinct categories: stable prostate, slow drift, persistent excursion, transient excursion, and high frequency excursions. The phantom and fiducial were imaged using 140 kVp, 0.63 mAs per image at 1 Hz for a 60 s monitoring period. Both stereoscopic and monoscopic 3D localization accuracies were assessed by comparison to the ground-truth obtained from the Linac log file. Imaging dose was also assessed, using optically stimulated luminescence dosimeter inserts in the phantom. Stereoscopic localization accuracy varied between 0.13 ± 0.05 and 0.33 ± 0.30 mm, depending on the motion trajectory. Monoscopic localization accuracy varied from 0.2 ± 0.1 to 1.1 ± 0.7 mm. The largest localization errors were typically observed in the left-right direction. There were significant differences in accuracy between the two monoscopic views, but which view was better varied from trajectory to trajectory. The imaging dose was measured to be between 2 and 15 μGy/mAs, depending on location in the phantom. The authors have demonstrated the first use of monoscopic localization for a room-mounted dual x-ray system. Three-dimensional position estimation from monoscopic imaging permits continuous, uninterrupted intrafraction motion monitoring even in the presence of gantry rotation, which may block kV sources or imagers. This potentially allows for more accurate treatment delivery, by ensuring that the prostate does not deviate substantially from the initial setup position.


Hierarchical "tube-on-fiber" carbon/mixed-metal selenide nanostructures for high-performance hybrid supercapacitors.
This work reports hierarchical "tube-on-fiber" nanostructures, composed of carbon nanotubes (CNTs) on carbon nanofibers (CNFs), impregnated with mixed-metal selenide nanoparticles (Co-Zn-Se@CNTs-CNFs), as high performance supercapacitors. Co-Zn hybrid zeolitic imidazolate framework-67 (Co-Zn ZIF-67) was electrospun with polyacrylonitrile (PAN) to form nanofibers that were sequentially thermally treated and subjected to selenylation. The "tube-on-fiber" structure is designed to confine the Co-Zn mixed-metal selenide nanoparticles and prevents their agglomeration. Extruded CNTs rooting in carbon nanofibers further improve the electronic conductivity. The mixed-metal selenide allows more accommodation space and faradic reactions compared to single metal selenide. Based on these merits, the hierarchical Co-Zn-Se@CNTs-CNFs exhibit a high specific capacity of 1040.1 C g-1 (1891 F g-1) at 1 A g-1 with impressive rate performance in supercapacitors. Furthermore, a hybrid supercapacitor with Co-Zn-Se@CNTs-CNFs as the cathode and porous carbon nanofibers as the anode (denoted as Co-Zn-Se@CNTs-CNFs//PCNFs) is fabricated. It delivers a superior energy and power density of 61.4 W h kg-1 and 754.4 W kg-1, respectively, and meanwhile retains 31.7 W h kg-1 of the energy density with 15 421.6 W kg-1 of the working power. In addition, the assembled supercapacitor device displays an excellent capacity retention of 88.6% after 8000 cycles at 5 A g-1.


Structural principles of distinct assemblies of the human α4β2 nicotinic receptor.
Fast chemical communication in the nervous system is mediated by neurotransmitter-gated ion channels. The prototypical member of this class of cell surface receptors is the cation-selective nicotinic acetylcholine receptor. As with most ligand-gated ion channels, nicotinic receptors assemble as oligomers of subunits, usually as hetero-oligomers and often with variable stoichiometries 1 . This intrinsic heterogeneity in protein composition provides fine tunability in channel properties, which is essential to brain function, but frustrates structural and biophysical characterization. The α4β2 subtype of the nicotinic acetylcholine receptor is the most abundant isoform in the human brain and is the principal target in nicotine addiction. This pentameric ligand-gated ion channel assembles in two stoichiometries of α- and β-subunits (2α:3β and 3α:2β). Both assemblies are functional and have distinct biophysical properties, and an imbalance in the ratio of assemblies is linked to both nicotine addiction2,3 and congenital epilepsy4,5. Here we leverage cryo-electron microscopy to obtain structures of both receptor assemblies from a single sample. Antibody fragments specific to β2 were used to 'break' symmetry during particle alignment and to obtain high-resolution reconstructions of receptors of both stoichiometries in complex with nicotine. The results reveal principles of subunit assembly and the structural basis of the distinctive biophysical and pharmacological properties of the two different stoichiometries of this receptor.


The effects of benzoflavones on polycyclic hydrocarbon metabolism and skin tumor initiation.
The effects of benzoflavones on skin tumor initiation by polycyclic hydrocarbons and epidermal aryl hydrocarbon hydroxylase were investigated. 7,8-Benzoflavone (7,8-BF) was found to be a potent inhibitor of the inhibition of skin tumors by 3-methylcholanthrene (MC) as well as 7,12-dimethylbenz(a)anthracene (DMBA). 5,6-Benzoflavone(5,6-BF) inhibited tumor initiation by MC and DMBA, but to a lesser degree than 7,8-BF. Dose-response studies of the capacity of 7,8-BF to inhibit DMBA tumor initiation revealed that 7,8-BF was an effective inhibitor at 2.5 microgram and a maximum inhibition of 90% occurred at 100 microgram of 7,8-FB. The tumor initiating ability of 7-hydroxymethyl-12-methylbenz(a)anthracene (7-OHMe-12MeBA) was not inhibited by 7,8-BF. Epidermal aryl hydrocarbon(benzo(a)pyrene hydroxylase(AHH) was increased by 5,6-BF and either had no effect or was slightly inhibited by 7,8-BF when given either topically or i.p. Both flavones when added directly to the assay tubes inhibited the in vitro epidermal AHH activity from control and MC pretreated mice by greater than 75%. When added in vitro, 7,8-BF and 5,6-BF inhibited epidermally mediated covalent binding of radioactive DMBA and dibenz(a,h)anthracene to DNA by 50% or more. The inhibition of skin tumor initiation by 7,8-BF and 5,6-BF appears to be partially related to its ability to inhibit the formation of electrophilic intermediates.


The Effect of Automated Mammogram Orders Paired With Electronic Invitations to Self-schedule on Mammogram Scheduling Outcomes: Observational Cohort Comparison.
Screening mammography is recommended for the early detection of breast cancer. The processes for ordering screening mammography often rely on a health care provider order and a scheduler to arrange the time and location of breast imaging. Self-scheduling after automated ordering of screening mammograms may offer a more efficient and convenient way to schedule screening mammograms. The aim of this study was to determine the use, outcomes, and efficiency of an automated mammogram ordering and invitation process paired with self-scheduling. We examined appointment data from 12 months of scheduled mammogram appointments, starting in September 2019 when a web and mobile app self-scheduling process for screening mammograms was made available for the Mayo Clinic primary care practice. Patients registered to the Mayo Clinic Patient Online Services could view the schedules and book their mammogram appointment via the web or a mobile app. Self-scheduling required no telephone calls or staff appointment schedulers. We examined uptake (count and percentage of patients utilizing self-scheduling), number of appointment actions taken by self-schedulers and by those using staff schedulers, no-show outcomes, scheduling efficiency, and weekend and after-hours use of self-scheduling. For patients who were registered to patient online services and had screening mammogram appointment activity, 15.3% (14,387/93,901) used the web or mobile app to do either some mammogram self-scheduling or self-cancelling appointment actions. Approximately 24.4% (3285/13,454) of self-scheduling occurred after normal business hours/on weekends. Approximately 9.3% (8736/93,901) of the patients used self-scheduling/cancelling exclusively. For self-scheduled mammograms, there were 5.7% (536/9433) no-shows compared to 4.6% (3590/77,531) no-shows in staff-scheduled mammograms (unadjusted odds ratio 1.24, 95% CI 1.13-1.36; P<.001). The odds ratio of no-shows for self-scheduled mammograms to staff-scheduled mammograms decreased to 1.12 (95% CI 1.02-1.23; P=.02) when adjusted for age, race, and ethnicity. On average, since there were only 0.197 staff-scheduler actions for each finalized self-scheduled appointment, staff schedulers were rarely used to redo or "clean up" self-scheduled appointments. Exclusively self-scheduled appointments were significantly more efficient than staff-scheduled appointments. Self-schedulers experienced a single appointment step process (one and done) for 93.5% (7553/8079) of their finalized appointments; only 74.5% (52,804/70,839) of staff-scheduled finalized appointments had a similar one-step appointment process (P<.001). For staff-scheduled appointments, 25.5% (18,035/70,839) of the finalized appointments took multiple appointment steps. For finalized appointments that were exclusively self-scheduled, only 6.5% (526/8079) took multiple appointment steps. The staff-scheduled to self-scheduled odds ratio of taking multiple steps for a finalized screening mammogram appointment was 4.9 (95% CI 4.48-5.37; P<.001). Screening mammograms can be efficiently self-scheduled but may be associated with a slight increase in no-shows. Self-scheduling can decrease staff scheduler work and can be convenient for patients who want to manage their appointment scheduling activity after business hours or on weekends.


GEOPHYSICS: Atmosphere Drives Earth's Tipsiness.
For more than a century, geophysicists who track Earth's rotation have sensed a rhythmic unsteadiness about the planet, an ever-so-slight wobbling whose source remained frustratingly mysterious. But researchers have been homing in on the roots of the so-called Chandler wobble, and now a report in the 1 August issue of Geophysical Research Letters fingers the shifting pressures of the deep sea and ultimately the fickle winds of the atmosphere.


Limitations in a frataxin knockdown cell model for Friedreich ataxia in a high-throughput drug screen.
Pharmacological high-throughput screening (HTS) represents a powerful strategy for drug discovery in genetic diseases, particularly when the full spectrum of pathological dysfunctions remains unclear, such as in Friedreich ataxia (FRDA). FRDA, the most common recessive ataxia, results from a generalized deficiency of mitochondrial and cytosolic iron-sulfur cluster (ISC) proteins activity, due to a partial loss of frataxin function, a mitochondrial protein proposed to function as an iron-chaperone for ISC biosynthesis. In the absence of measurable catalytic function for frataxin, a cell-based assay is required for HTS assay. Using a targeted ribozyme strategy in murine fibroblasts, we have developed a cellular model with strongly reduced levels of frataxin. We have used this model to screen the Prestwick Chemical Library, a collection of one thousand off-patent drugs, for potential molecules for FRDA. The frataxin deficient cell lines exhibit a proliferation defect, associated with an ISC enzyme deficit. Using the growth defect as end-point criteria, we screened the Prestwick Chemical Library. However no molecule presented a significant and reproducible effect on the proliferation rate of frataxin deficient cells. Moreover over numerous passages, the antisense ribozyme fibroblast cell lines revealed an increase in frataxin residual level associated with the normalization of ISC enzyme activities. However, the ribozyme cell lines and FRDA patient cells presented an increase in Mthfd2 transcript, a mitochondrial enzyme that was previously shown to be upregulated at very early stages of the pathogenesis in the cardiac mouse model. Although no active hit has been identified, the present study demonstrates the feasibility of using a cell-based approach to HTS for FRDA. Furthermore, it highlights the difficulty in the development of a stable frataxin-deficient cell model, an essential condition for productive HTS in the future.


beta(2)-Adrenoceptor activation attenuates endotoxin-induced acute renal failure.
Abnormalities in the beta(2)-adrenergic control of organ function have been implicated in the pathogenesis of several disease states, such as septic shock. The objectives of the present study were to define the contribution of beta(2)-adrenoceptors (beta(2)-AR) to normal renal physiology and to investigate whether overexpression of renal beta(2)-AR might be potentially beneficial in preventing progressive renal damage associated with endotoxemia. Adenoviral transgenes containing the human beta(2)-AR (Adeno-beta(2)-AR) were constructed and delivered into the rat kidney by means of intraparenchymal injections. Administration of 10(9) total viral particles of Adeno-beta(2)-AR induced an approximately threefold increase in beta(2)-AR density in the renal tissue, which 2 wk after delivery, enhanced GFR and sodium reabsorption compared with control rats. The enhanced GFR was abolished by the addition of the beta(2)-AR antagonist, ICI 118,551. Administration of lipopolysaccharide (LPS) caused a reduction in GFR, beta(2)-AR density, and cAMP together with enhanced TNF-alpha mRNA in the kidney. In rats overexpressing beta(2)-AR, the reduction in baseline GFR and elevation of TNF-alpha mRNA and leukocyte infiltration into the kidney associated with the endotoxin were blocked. These findings suggested the possibility that a renal-specific overexpression of beta(2)-AR preserves basal renal function in response to a ligand-independent beta(2)-AR activation and that the delivery of Adeno-beta(2)-AR gene is a potential novel therapeutic strategy for treatment of acute renal failure associated with sepsis.


Simulated scaphoid proximal pole fracture.
Five fresh cadaver upper extremities were studied with use of a static positioning frame, pressure-sensitive film, a microcomputer-based videodigitizing system, and a Sun station image analysis system to assess the load bearing characteristics of the scaphoid in the proximal carpal joint. Specimens were studied in their normal condition, after a proximal pole osteotomy of the scaphoid, and after resection of the proximal pole of the scaphoid. The amount of contact area born through the scaphoid fossa was essentially the same whether the scaphoid was intact, or after a simulated scaphoid fracture of its proximal pole, or after resection of the proximal pole. The scaphoid contact area and pressure, although overall relatively constant, was redistributed after osteotomy, resulting in increased contact area under the distal fragment and no change or a slight decrease in the contact area under the proximal fragment of the scaphoid. After resection of the proximal fragment, all scaphoid contact area and pressure was born by the distal scaphoid fragment. The contact area and pressure characteristics of the lunate remained unchanged in all conditions compared with the normal condition. There were no significant changes in the locations of the centroids of the scaphoid segments and the lunate in any of the conditions tested.


Uptake and metabolism of beta-carotene and retinal by C3H/10T1/2 cells.
Both beta-carotene (beta-C), a vitamin A precursor, and vitamin A itself have been shown to reversibly inhibit neoplastic transformation in 10T1/2 cells during the progression phase of carcinogenesis. In order to determine whether the activity of beta-C in these cells may be attributed to conversion to vitamin A or is intrinsic to the carotenoid molecule, the uptake and metabolism of beta-C, and of retinal, the immediate product of dioxygenase-cleavage of beta-C, was studied in 10T1/2 cells. Cellular uptake of 2.6 nmol/10(6) cells occurred 24 h after treatment with 10(-5) M beta-C. Thereafter, cell levels remained relatively stable between 1 and 2 nmol/10(6) cells over the 1-week treatment period. Upon removal of beta-C from the medium, cellular levels decreased by approximately 80% in 2 weeks, then stabilized. Retinal was rapidly and quantitatively converted to retinol by 10T1/2 cells, suggesting that the inhibitory action of retinal on neoplastic transformation in these cells is due to its conversion to retinol, and that any enzymatic conversion of beta-C to retinal by these cells would be expected to result in retinol as the end product. Using [14C]beta-C, we found no evidence for formation of [14C]retinol, [14C]retinal or [14C]retinoic acid using sensitive HPLC. We therefore conclude that beta-C has intrinsic chemopreventive activity in 10T1/2 cells, perhaps due to its anti-oxidant properties.


Mortality rates, hospital length of stay, and the cost of treating subarachnoid hemorrhage in older patients: institutional and geographical differences.
The risk of disability and death and the cost of medical care are particularly high for patients with aneurysmal subarachnoid hemorrhage (SAH) who are 65 years of age or older. A retrospective analysis of 47,408 Medicare patients treated over an 8-year period was performed to determine whether a relationship exists between the mortality rate and surgical volume for older patients with SAH. The mortality rate, length of stay in the hospital, and cost of treatment for patients with SAH in California and New York state were also compared. The mortality rate was 14.3% for patients with SAH who were 65 years old or older and who were treated surgically in hospitals in which an average of five or more craniotomies were performed per year; in hospitals averaging between one and five craniotomies annually the mortality rate was 18.4%; and in those averaging less than one such operation per year the rate was 20.5% (trend p = 0.01). There was no difference in the mortality rate for patients in California versus the rate for those in New York. Surgically and medically treated patients, respectively, left the hospital an average of 6.7 and 5.1 days sooner in California than in New York. The unadjusted average reimbursement from Medicare to hospitals for surgically treated patients averaged $1468 more in New York than in California (p < 0.0001), but was equivalent for medically treated patients in the two states. The mortality rate in older patients who are treated surgically for SAH may be inversely correlated with the annual number of craniotomies performed for SAH in patients 65 years of age or older at a given institution. Hospital stays for patients with SAH are significantly shorter in California than in New York.


A streamlined DNA tool for global identification of heavily exploited coastal shark species (genus Rhizoprionodon).
Obtaining accurate species-specific landings data is an essential step toward achieving sustainable shark fisheries. Globally distributed sharpnose sharks (genus Rhizoprionodon) exhibit life-history characteristics (rapid growth, early maturity, annual reproduction) that suggests that they could be fished in a sustainable manner assuming an investment in monitoring, assessment and careful management. However, obtaining species-specific landings data for sharpnose sharks is problematic because they are morphologically very similar to one another. Moreover, sharpnose sharks may also be confused with other small sharks (either small species or juveniles of large species) once they are processed (i.e., the head and fins are removed). Here we present a highly streamlined molecular genetics approach based on seven species-specific PCR primers in a multiplex format that can simultaneously discriminate body parts from the seven described sharpnose shark species commonly occurring in coastal fisheries worldwide. The species-specific primers are based on nucleotide sequence differences among species in the nuclear ribosomal internal transcribed spacer 2 locus (ITS2). This approach also distinguishes sharpnose sharks from a wide range of other sharks (52 species) and can therefore assist in the regulation of coastal shark fisheries around the world.


Italian translation, cultural adaptation, and pilot testing of a questionnaire to assess family burden in inherited ichthyoses.
Inherited ichthyoses are rare disorders characterized by generalized skin scaling. Among them, autosomal recessive congenital ichthyoses (ARCI) form a major subgroup presenting lifelong and severely disabling cutaneous and extracutaneous features and symptoms for which no curative treatment is available. Management relies on daily time-consuming and distressing topical medications. Disease manifestations, symptoms, and daily care affect not only the patient self-perception, but also different dimensions of patient and family life. To date, there is only a French validated ichthyosis-specific questionnaire, "Family Burden in Ichthyosis" (FBI), for the evaluation of family disease burden. It addresses economical aspects, daily life, familial and personal relationships, work, and psychological impact. The aim of our study was to develop an Italian translation of the French FBI questionnaire and to pilot-test it in ARCI patients. The guidelines for cross-cultural adaptation of health-related quality of life measures were followed. Specifically, two independent forward translations were produced, followed by a reconciliation step by a multidisciplinary expert committee and back-translation. Revision of the original text and all translations was performed by the expert committee leading to a final version, which was pilot-tested by cognitive debriefing on 10 caregivers whose comments were evaluated by the committee. The translation and reconciliation process led to minor changes in five items in order to clarify the questions in relation to the possible answers or to obtain semantic/idiomatic/cultural equivalence of the Italian version with the French one. The cognitive debriefing process resulted into further minor wording modifications in four items to describe more precisely the disease impact according to parents' comments. The FBI developer approved the final Italian FBI version. The Italian version of the FBI generated in the present study is a useful instrument to measure the impact of ichthyosis on family daily life, education and working activities, psychological implications, and the disease economic load. The questionnaire will be further validated through a multicenter Italian study on burden of ARCI. A validated Italian questionnaire is a valuable tool for future clinical trials. In addition, it can be used to rapidly identify family distressing situations, which require attention and prompt intervention.


High incidence of occult familial SDHD cases amongst Czech patients with head and neck paragangliomas.
Head and neck paragangliomas (HNPGLs) are rare neuroendocrine tumors, which are mostly benign in nature. Amongst all genes, Succinate Dehydrogenase Subunit D (SDHD) is the most commonly mutated in familial HNPGLs. In about 30% of HNPGLs, germline mutations in SDHD can also occur in the absence of positive family history, thus giving rise to "occult familial" cases. Our aim was to evaluate the pattern of SDHD germline mutations in Czech patients with HNPGLs. We analyzed a total of 105 patients with HNPGLs from the Otorhinolaryngology departments of 2 tertiary centers between 2006 - 2021. All underwent complex diagnostic work-up and were also consented for genetic analysis. Eighty patients aged 13-76 years were included; around 60% with multiple PGLs were males. Carotid body tumor was the most frequently diagnosed tumor. Germline SDHD mutation was found in only 12% of the Czech patients; approximately 78% of those harboring the mutation had negative family history. The mutation traits had higher affiliation for multiple tumors with nearly 70% patients of ≤ 40 years of age. An SDHD mutation variant was shared amongst unrelated patients but no founder-effect was established. Our findings confirmed that the pattern of SDHD mutation distribution amongst HNPGLs in Czech Republic differs from most studies worldwide.


A Conserved DNA Repeat Promotes Selection of a Diverse Repertoire of Trypanosoma brucei Surface Antigens from the Genomic Archive.
African trypanosomes are mammalian pathogens that must regularly change their protein coat to survive in the host bloodstream. Chronic trypanosome infections are potentiated by their ability to access a deep genomic repertoire of Variant Surface Glycoprotein (VSG) genes and switch from the expression of one VSG to another. Switching VSG expression is largely based in DNA recombination events that result in chromosome translocations between an acceptor site, which houses the actively transcribed VSG, and a donor gene, drawn from an archive of more than 2,000 silent VSGs. One element implicated in these duplicative gene conversion events is a DNA repeat of approximately 70 bp that is found in long regions within each BES and short iterations proximal to VSGs within the silent archive. Early observations showing that 70-bp repeats can be recombination boundaries during VSG switching led to the prediction that VSG-proximal 70-bp repeats provide recombinatorial homology. Yet, this long held assumption had not been tested and no specific function for the conserved 70-bp repeats had been demonstrated. In the present study, the 70-bp repeats were genetically manipulated under conditions that induce gene conversion. In this manner, we demonstrated that 70-bp repeats promote access to archival VSGs. Synthetic repeat DNA sequences were then employed to identify the length, sequence, and directionality of repeat regions required for this activity. In addition, manipulation of the 70-bp repeats allowed us to observe a link between VSG switching and the cell cycle that had not been appreciated. Together these data provide definitive support for the long-standing hypothesis that 70-bp repeats provide recombinatorial homology during switching. Yet, the fact that silent archival VSGs are selected under these conditions suggests the 70-bp repeats also direct DNA pairing and recombination machinery away from the closest homologs (silent BESs) and toward the rest of the archive.