Validation of a homology model of Mycobacterium tuberculosis DXS: rationalization of observed activities of thiamine derivatives as potent inhibitors of two orthologues of DXS.
The enzyme DXS catalyzes the first, rate-limiting step of the 2-C-methyl-d-erythritol-4-phosphate (MEP, 1) pathway using thiamine diphosphate (ThDP) as cofactor; the DXS-catalyzed reaction constitutes also the first step in vitamin B1 and B6 metabolism in bacteria. DXS is the least studied among the enzymes of this pathway in terms of crystallographic information, with only one complete crystal structure deposited in the Protein Data Bank (Deinococcus radiodurans DXS, PDB: ). We synthesized a series of thiamine and ThDP derivatives and tested them for their biochemical activity against two DXS orthologues, namely D. radiodurans DXS and Mycobacterium tuberculosis DXS. These experimental results, combined with advanced docking studies, led to the development and validation of a homology model of M. tuberculosis DXS, which, in turn, will guide medicinal chemists in rationally designing potential inhibitors for M. tuberculosis DXS.


Using a parental checklist to identify diagnostic groups in children with communication impairment: a validation of the Children's Communication Checklist--2.
The Children's Communication Checklist (CCC 1998) was revised in 2003 (CCC-2) to provide a general screen for communication disorder and to identify pragmatic/social interaction deficits. Two validation studies were conducted with different populations of children with language and communication impairments. In Study 1, the questionnaire was given to families of 87 children attending full-time special education for specific language impairment, pragmatic language impairments or autistic spectrum disorders. In addition, the teachers of half the sample completed CCC-2 forms for the same children, providing evidence for interrater agreement. In Study 2, the sample was increased to include 24 children with similar diagnoses in educational contexts in Scotland and 27 children referred for clinical evaluation at a neurodevelopment centre. The CCC-2 distinguished children with communication impairments from non-impaired peers. Furthermore, the social-interaction deviance composite (SIDC) of the CCC-2 identified children with disproportionate pragmatic and social difficulties in relation to their structural language impairments. This measure also had good interrater agreement (r=0.79). CCC-2 provides a useful screening measure for communication impairment and can be helpful in identifying children who should be referred for more detailed assessment of possible autistic spectrum disorder. However, the present data highlight substantial overlap amongst groups with 'distinct' diagnoses. It is suggested that it is unrealistic to use the CCC-2 to make categorical distinctions on this continuum of disorder.


Prevalence and predictors of low future expectations among Syrian refugees resettled in Sweden.
Future Expectation is important for motivation and wellbeing, however drastic life events such as in refugee situations may result in low expectations. This study aims to investigate the prevalence and determinants of low future expectations among Syrian refugees resettled in Sweden. A random sample of 1215 Syrian refugees resettled in Sweden responded to questionnaire. Weighted analyses and adjusted relative risks were conducted to determine the prevalences and predictors of low future expectations. Synergy index was calculated for low social support and depression in relation to low expectations. The prevalences of low future expectations for labour market, social and economic intergration were 10.9%, 13.4% and 14.1% respectively. Longer stay in Sweden, being older, low social support and depression were associated with low future expectations. The simultaneous presence of depression and low social support had a synergistic effect on low social expectation. Understanding and addressing factors related to low future expectations among refugees may be useful for facilitating their labour market, social and economic integration.


Dyserythropoiesis of myelodysplastic syndromes.
Myelodysplastic syndromes (MDS) are heterogeneous diseases of the hematopoietic stem cell in the elderly. Anemia is the main symptom that mostly correlates with dysplastic erythropoiesis in the bone marrow. We will review the recent advances in understanding the diverse mechanisms of dyserythropoiesis. Dyserythropoiesis defined as 10% dysplastic erythroid cells in the bone marrow is found in more than 80% of early MDS. Immature erythroblasts accumulate at the expense of mature erythroblasts due to differentiation arrest and apoptosis. In early MDS with dyserythropoiesis, caspase-dependent cleavage of the erythroid transcription factor GATA-1 occurring in basophilic erythroblasts accounts for impairment of final maturation. Depending on initiating genetic alteration, specific mechanisms contribute to erythroid defect. In MDS with 5q deletion, the haploinsufficiency of ribosomal protein gene, RPS14, opposes the transition of immature to mature erythroblasts by inducing a p53-dependent ribosome stress, cell cycle arrest and apoptosis. Recent work identifies the activation of a p53-S100A8/9 innate immune pathway that both intrinsically and extrinsically contributes to defective erythropoiesis. In MDS with ring sideroblasts, a paradigm of dyserythropoiesis, a unique mutation in SF3B1 splicing factor gene induces a multiplicity of alterations at RNA level that deeply modifies the patterns of gene expression. Insights in the pathophysiology of MDS with dyserythropoiesis may guide the choice of the appropriate therapy, for instance lenalidomide in MDS with del(5q). A better understanding of the mechanisms of dyserthropoiesis is required to treat anemia in non-del(5q) MDS, especially in case of resistance to first-line therapy by erythropoiesis-stimulating agents.


Radiation Dose and Procedure Time for 994 CT-guided Spine Pain Control Procedures.
Image guidance for spine pain control procedures, including epidural steroid injection, nerve root block, and facet block, can be performed with either computed tomography (CT) or conventional fluoroscopy. CT has the advantage of improved anatomic localization and use of air for contrast; however, there are concerns that CT leads to higher radiation dose and longer procedure time. To evaluate procedure time and radiation dose for multiple types of spine pain control procedures performed under CT guidance. Retrospective evaluation. Department of radiology in single academic medical center. Institutional review board approval was obtained. We reviewed CT-guided spine procedures performed over a 12-month period from January 2012 to December 2012. Procedure type, procedure time, and dose-length product were recorded. Patient age and gender were recorded for each case; additionally, demographic and medical history data were obtained for a sub-group of patients. Nine hundred ninety-four studies (performed in 699 patients) were reviewed, including 585 epidural steroid injections, 228 nerve root blocks, and 90 facet blocks. For all studies, procedure time averaged 7:34 ± 5:05, and dose-length product averaged 75 mGy·cm ± 61. Additional medical history (available for 483 patients) revealed high rate of obesity (body mass index [BMI] = 30 ± 6.8, with 76% of patients overweight [BMI > 25] and 42% obese [BMI > 30]), and frequent medical comorbidities (including hypertension [n = 179], diabetes [n = 101], renal failure [n = 30], and heart failure [n = 17]). This study was performed retrospectively, and limited to a single institution. These findings add to the growing evidence that CT guidance is a safe and effective technique for epidural steroid injection. These results further demonstrate that other spine intervention procedures, including nerve root block and facet block, can also be performed under CT guidance with short procedure time and reasonable levels of radiation exposure. This approach can be effectively used in a patient population with a high rate of obesity and medical comorbidities.


Measuring the effects of neurodevelopmental treatment on the daily living skills of 2 children with cerebral palsy.
This study examined the short-term effects of neurodevelopmental treatment (NDT) on the motor performance of daily living skills in 2 girls with cerebral palsy, aged 27 and 32 months. For 12 weeks, the subjects received treatment sessions consisting of NDT followed by play, or vice versa. After each treatment of play or NDT, the subjects were videotaped performing individually prescribed dressing activities. Trained raters, unaware of the type of treatment given, scored the videotapes on ordinally scaled dressing criteria. The results were not statistically significant for NDT or play for any criteria. The importance of this study is its development and exploration of refinements in single-subject research methods that show promise for use in the documentation of treatment outcomes. Such refinements include the use of functional activity as an evaluation tool and ordinal scales for the assessment of change.


Are airflow obstruction and radiographic evidence of emphysema risk factors for lung cancer? A nested case-control study using quantitative emphysema analysis.
Several studies have identified airflow obstruction as a risk factor for lung cancer independent of smoking history, but the risk associated with the presence of radiographic evidence of emphysema has not been extensively studied. We proposed to assess this risk using a quantitative volumetric CT scan analysis. Sixty-four cases of lung cancer were identified from a prospective cohort of 1,520 participants enrolled in a spiral CT scan lung cancer screening trial. Each case was matched to six control subjects for age, sex, and smoking history. Quantitative CT scan analysis of emphysema was performed. Spirometric measures were also conducted. Data were analyzed using conditional logistic regression making use of the 1:6 set groups of 64 cases and 377 matched control subjects. Decreased FEV(1) and FEV(1)/FVC were significantly associated with a diagnosis of lung cancer with ORs of 1.15 (95% CI, 1.00-1.32; P = .046) and 1.29 (95% CI, 1.02-1.62; P = .031), respectively. The quantity of radiographic evidence of emphysema was not found to be a significant risk for lung cancer with OR of 1.042 (95% CI, 0.816-1.329; P = .743). Additionally, there was no significant association between severe emphysema and lung cancer with OR of 1.57 (95% CI, 0.73-3.37). We confirm previous observations that airflow obstruction is an independent risk factor for lung cancer. The absence of a clear relationship between radiographic evidence of emphysema and lung cancer using an automated quantitative volumetric analysis may result from different population characteristics than those of prior studies, radiographic evidence of emphysema quantitation methodology, or absence of any relationship between emphysema and lung cancer risk.


[Medin modular implant for total knee arthroplasty--mid-term results].
PURPOSE OF THE STUDY To evaluate the mid-term results of total knee arthroplasty (TKA) using a Medin Modular joint prosthesis in a standard design preserving the posterior cruciate ligament. MATERIAL The study comprised 206 consecutive total knee arthroplasties, 10 bilateral, 60 in men and 136 in women. Twelve patients died of causes unrelated to the surgery and 14 knee replacements were lost to follow-up. A total of 180 were evaluated at an average follow-up of 6.5 years (range, 5 to 8.7 years). The patients ranged in age from 43 to 84 years (average, 69.5 years). The basic indication for surgery was osteoarthritis in 179 and rheumatoid arthritis in 37 cases. METHODS Pain, range of motion, stability and function were scored on a 100-point scale of the Hospital for Special Surgery (HSS) Scoring System. Subjective satisfaction was essessed by the patient's willingness to undergo the surgery again. At yearly intervals, radiographs in two projections were examined for radiolucent lines around the implant. The Kaplan-Meier survival curve was evaluated for the whole group and compared with that for non-infected cases only. The mechanical properties of the implant were most reliably shown by the survival curve for the subgroup of knee replacements free of infection. RESULTS At the final follow-up out of 180 knees, 169 were fully functioning and 11 required revision arthroplasty for the following reasons: polyethylene wear, one knee; instability, two; infection, six; patellar fracture, one; and problems associated with the femoro-patellar joint, one knee. The Kaplan-Meier curve for the whole group showed survival of 92 % of the implants at the end of the 9-years follow-up time. In the infection-free subgroup, in which aseptic loosenings were only included, there was a 95 % survival. The average range of flexion was over 106 degrees, and only 11 knees had flexion below 90 degree lack of. Extension from 5 to 10 degrees was found in four knees. Thirteen knees showed instability that did not interfere with function. In 191 knees, after surgery alignment was optimal, between 0 and 5-degree valgus; in 11 knees varus up to 5 degrees and valgus up to 10 degrees were found. No pain was recorded in 123 knees, occasional pain with no effect on function in 36 cases, and moderate pain after weight-bearing in 10 knees. By the HSS score, 136 knees had excellent and very good (over 90 points) outcomes and 21 had unsatisfactory or poor outcomes (below 85 points); the remaining 23 had satisfactory outcomes (between 89 and 85 points). The subjective evaluation was positive for 156 knees. Of 169 TKA evaluated radiographs, 141 were without signs of radiolucency, 17 showed a slight marginal radiolucent line and 11 had a more pronounced radiolucent line up to 1 mm in width, but without progression. DISCUSSION A total of 93 % of total replacements were evaluated and 7 % were lost to follow-up. Therefore, the number of remaining patients is big enough to make the group valid for assessment. The rate of complications, including 2.9 % of infected knees, is in agreement with the reported values. The occurence of polyethylene wear was negligible. A comparison of the cumulative survival curve for the whole group with that for the infection-free subgroup showed that the effect of infection on the TKA outcome was statistically significant (p=0.0431), and that late infection remains the most serious complication. Aseptic loosening plays a less important role in implant survival evaluated at mid-term intervals. CONCLUSIONS The rate of complications for Medin Modular implants evaluated at an average follow-up of 6.5 years is not different from the values reported for similar prostheses. The most serious complication is late infection. Persistent pain is usually located to the femoro-patellar articulation, but it seldom markedly affects joint function. The cumulative survival curve for the whole group was 92 % at a 9 years. The HSS evaluation of the whole group showed that 75 % of the knees achieved mor than 90 points.


Cognitive reorganization during pregnancy and the postpartum period: an evolutionary perspective.
Where the non-human animal research investigating reproduction-induced cognitive reorganization has focused on neural plasticity and adaptive advantage in response to the demands associated with pregnancy and parenting, human studies have primarily concentrated on pregnancy-induced memory decline. The current review updates Henry and Rendell's 2007 meta-analysis, and examines cognitive reorganization as the result of reproductive experience from an adaptationist perspective. Investigations of pregnancy-induced cognitive change in human females may benefit by focusing on areas, such as social cognition, where a cognitive advantage would serve a protective function, and by extending the study duration beyond pregnancy into the postpartum period.


Genitals and ethnicity: the politics of genital modifications.
The discrepancy in societal attitudes toward female genital cosmetic surgery for European women and female genital cutting in primarily African girl children and women raises the following fundamental question. How can it be that extensive genital modifications, including reduction of labial and clitoral tissue, are considered acceptable and perfectly legal in many European countries, while those same societies have legislation making female genital cutting illegal, and the World Health Organization bans even the "pricking" of the female genitals? At present, tensions are obvious as regards the modification of female genitalia, and current legislation and medical practice show inconsistencies in relation to women of different ethnic backgrounds. As regards the right to health, it is questionable both whether genital cosmetic surgery is always free of complications and whether female genital cutting always leads to them. Activists, national policymakers and other stakeholders, including cosmetic genital surgeons, need to be aware of these inconsistencies and find ways to resolve them and adopt non-discriminatory policies. This is not necessarily an issue of either permitting or banning all forms of genital cutting, but about identifying a consistent and coherent stance in which key social values - including protection of children, bodily integrity, bodily autonomy, and equality before the law - are upheld.


Speckle size of light scattered from slightly rough cylindrical surfaces.
This research is an extension of the optical method of quality control presented in a previous paper [Appl. Opt. 39, 5811 (2000)] to the case of slightly rough cylindrical surfaces. Applying the Kirchhoff scalar diffraction theory yields an analytical expression of the autocorrelation function of the intensity scattered from slightly rough cylindrical surfaces. This function, which is related to speckle size and shape, is shown to depend on the surface correlation length, unlike for plane surfaces for which the speckle depends on the illuminated area only. The theoretical expression is compared with that for the speckle produced by the light scattered from a cylindrical bearing and from various high-quality wires, showing that the method allows the correlation lengths of high-quality cylindrical surfaces to be determined.


Magnetic field sensor using tilted fiber grating interacting with magnetic fluid.
A novel magnetic field sensor using tilted fiber Bragg grating (TFBG) interacting with magnetic fluid is proposed and experimentally demonstrated. The TFBG is surrounded by magnetic fluid whose complex refractive index changes with external magnetic field. The guiding properties of cladding modes excited by the TFBG are therefore modulated by the external magnetic field. As a result, the magnetic field strength measurement is successfully achieved within a range up to 196 Gauss by monitoring extinction ratio of cladding mode resonance. Furthermore, temperature variation can be obtained simultaneously from the wavelength shift of the TFBG transmission spectrum.


S100A12 Expression Is Modulated During Monocyte Differentiation and Reflects Periodontitis Severity.
S100A12 is a calcium-binding protein of the S100 subfamily of myeloid-related proteins that acts as an alarmin to induce a pro-inflammatory innate immune response. It has been linked to several chronic inflammatory diseases, however its role in the common oral immunopathology periodontitis is largely unknown. Previous in vitro monoculture experiments indicate that S100A12 production decreases during monocyte differentiation stages, while the regulation within tissue is poorly defined. This study evaluated S100A12 expression in monocyte subsets, during monocyte-to-macrophage differentiation and following polarization, both in monoculture and in a tissue context, utilizing a three-dimensional co-culture oral tissue model. Further, we explored the involvement of S100A12 in periodontitis by analyzing its expression in peripheral circulation and gingival tissue, as well as in saliva. We found that S100A12 expression was higher in classical than in non-classical monocytes. S100A12 expression and protein secretion declined significantly during monocyte-to-macrophage differentiation, while polarization of monocyte-derived macrophages had no effect on either. Peripheral monocytes from periodontitis patients had higher S100A12 expression than monocytes from controls, a difference particularly observed in the intermediate and non-classical monocyte subsets. Further, monocytes from periodontitis patients displayed an increased secretion of S100A12 compared with monocytes from controls. In oral tissue cultures, monocyte differentiation resulted in increased S100A12 secretion over time, which further increased after inflammatory stimuli. Likewise, S100A12 expression was higher in gingival tissue from periodontitis patients where monocyte-derived cells exhibited higher expression of S100A12 in comparison to non-periodontitis tissue. In line with our findings, patients with severe periodontitis had significantly higher levels of S100A12 in saliva compared to non-periodontitis patients, and the levels correlated to clinical periodontal parameters. Taken together, S100A12 is predominantly secreted by monocytes rather than by monocyte-derived cells. Moreover, S100A12 is increased in inflamed tissue cultures, potentially as a result of enhanced production by monocyte-derived cells. This study implicates the involvement of S100A12 in periodontitis pathogenesis, as evidenced by increased S100A12 expression in inflamed gingival tissue, which may be due to altered circulatory monocytes in periodontitis.


Postcardioplegic myocardial recovery: effects of halothane, nifedipine, HOE 694, and quinacrine.
Halothane has been shown to be a powerful myocardial protectant during normothermic cardioplegic arrest and subsequent reperfusion. In view of its multiple effects on cellular Ca2+ movements and the role of this ion in ischemia-reperfusion injury, the questions of whether halothane is capable of maximally protecting the heart or whether combination therapy of halothane with other Ca2+ blocking agents may be more effective arose. Therefore, the effects of combination therapy with halothane and a calcium antagonist (nifedipine), or a Na+/H+ inhibitor (HOE 694), or a Na+/Ca2+ inhibitor (quinacrine) on postcardioplegic functional recovery were evaluated. The isolated perfused rat heart subjected to 45 minutes normothermic cardiac arrest was used as an experimental model. Dose-response curves were performed for each drug. Using the optimal dosage for each drug, the following results were obtained: (1) Nifedipine (10(-7) M; administered retrogradely 10 minutes before and after cardioplegia) and halothane (1.5% administered during cardioplegia), when administered separately, improved functional recovery. Combination therapy did not further improve protection. (2) HOE 694 (10(-7) M) or quinacrine (10(-9) M) improved post-cardioplegic functional recovery when added for 2 minutes at the onset of reperfusion. Simultaneous administration of HOE 694 and 1.5% halothane was the only combination that yielded additive protection. (3) Quinacrine, a phospholipase and Na+/Ca2+ exchanger inhibitor, appeared to be the most powerful drug used. In summary, the results obtained indicate that interventions aimed at preventing intracellular Ca2+ overload improve recovery after cardioplegic arrest. The beneficial effects of halothane could be further improved by HOE 694.


Pregnancy outcomes in women with a systemic right ventricle and transposition of the great arteries results from the ESC-EORP Registry of Pregnancy and Cardiac disease (ROPAC).
Cardiac disease is a major cause of maternal mortality. Data regarding pregnancy outcomes in women with a systemic right ventricle (sRV) are scarce. We studied pregnancy outcomes in women with an sRV after the atrial switch procedure for transposition of the great arteries (TGA) or congenitally corrected TGA (CCTGA). The ESC EORP Registry of Pregnancy and Cardiac Disease is an international prospective registry of pregnant women with cardiac disease. Pregnancy outcomes (maternal/fetal) in all women with an sRV are described. The primary end point was a major adverse cardiac event (MACE) defined as maternal death, supraventricular or ventricular arrhythmias requiring treatment, heart failure, aortic dissection, endocarditis, ischaemic coronary event and other thromboembolic events. Altogether, 162 women with an sRV (TGA n=121, CCTGA n=41, mean age 28.8±4.6 years) were included. No maternal mortality occurred. In 26 women, at least one MACE occurred, heart failure in 16 (9.8%), arrhythmias (atrial 5, ventricular 6) in 11 (6.7%) and others in 4 (2.5%). Prepregnancy signs of heart failure as well as an sRV ejection fraction <40% were predictors of MACE. One woman experienced fetal loss, while no neonatal mortality was observed. No significant differences were found between women with CCTGA and TGA. In the subset of women who had an echocardiogram before and after pregnancy, no clear deterioration in sRV was observed. The majority of women with an sRV tolerated pregnancy well with a favourable maternal and fetal outcome. Heart failure and arrhythmias were the most common MACE.


Deviation in the predefined calibration factors of the dose calibrators and the associated inaccuracy in the radioactivity measurements of beta-gamma emitters.
To determine whether the predefined calibration factors of the dose calibrators can provide accurate radioactivity measurements of beta-gamma emitters used in routine therapeutic nuclear medicine procedures. Two models of dose calibrators were used in the present study for radioactivity measurements of (153)Sm ethylenediamine-N, N, N', N'-tetrakis methylene phosphonic acid (EDTMP) and (177)Lu (EDTMP). A known (precalibrated) activity of each of the two beta emitters received by us from our National Supplier for administration to the patients with extensive bony metastases for bone pain palliation, was used for experiments. When we used the manufacturers' provided dial setting of 450 × 10, each of the dose calibrators underestimated the radioactivity of (177)Lu by about 9.0%. Dial settings of 403 × 10 and 408 × 10 for (177)Lu on CRC-15R and CRC-ultra dose calibrators respectively were calculated experimentally using an iterative approach. The radioactivity measurements made at these settings provided an excellent agreement with the specified values. Likewise, a dial setting of 230 for each of the two dose calibrators was calculated for (153)Sm, which provided a good agreement between the experimentally derived radioactivity values and the certified values. A deviation of ± 5.0% was observed when radioactivity of (177)Lu and (153)Sm was measured over a wide range (4.0 MBq to 2.1 GBq) for time intervals equivalent to 4.5 half-lives of each of the two radionuclides. A deviation of ± 5% was observed when radioactivity was counted in different dilution volumes and in syringes of varying size. These variations could lead to a cumulative error of about 20.0% toward the inaccuracy in the radioactivity measurements of the beta-gamma emitters and thus predefined calibration factors of the dose calibrators may require experimental re-setting of these parameters and periodic checking to provide accurate radioactivity estimates of beta-gamma emitters in a given clinical setting.


Low-dose risedronate sodium protects bone cells after abrupt oestrogen withdrawal.
To investigate the effects of low-dose risedronate sodium on the in vitro cellular profile of osteoblasts, adipocytes, osteocytes and osteoclasts in a rat model of abrupt oestrogen deficiency. Oestrogen deficiency was induced by ovariectomy in 24 female rats. The rats were treated with low-dose (0.24 μg/kg) or high-dose (2.4 μg/kg) risedronate sodium for 4 days presurgery, continuing every 3 days until 15 days postsurgery. Osteogenic and adipogenic differentiation were determined in cultured bone marrow cells by alkaline phosphatase and Oil Red O staining, respectively, and by osteogenic and adipogenic gene expression. Osteoclast formation was measured in bone marrow cells stimulated with macrophage colony-stimulating factor and receptor activator of nuclear factor κB ligand, and stained with tartrate-resistant acid phosphatase. Osteocyte apoptosis was assessed by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labelling assay and B-cell lymphoma-2 (Bcl-2) immunohistochemistry. Low-dose risedronate sodium enhanced osteoblast differentiation, suppressed adipocyte differentiation and osteoclast formation, and reduced osteocyte apoptosis through regulation of Bcl-2 and Bcl-2-associated X protein. Low-dose risedronate sodium may have clinical benefit in protecting against bone loss after abrupt oestrogen deficiency.


Hard X-ray detection using a single 100 nm diameter nanowire.
Submicron sized sensors could allow higher resolution in X-ray imaging and diffraction measurements, which are ubiquitous for materials science and medicine. We present electrical measurements of a single 100 nm diameter InP nanowire transistor exposed to hard X-rays. The X-ray induced conductance is over 5 orders of magnitude larger than expected from reported data for X-ray absorption and carrier lifetimes. Time-resolved measurements show very long characteristic lifetimes on the order of seconds, tentatively attributed to long-lived traps, which give a strong amplification effect. As a proof of concept, we use the nanowire to directly image an X-ray nanofocus with submicron resolution.


Cryogenic Media in Biomedical Applications: Current Advances, Challenges, and Future Perspectives.
This paper explores the crucial role of cryogenic mediums in driving breakthroughs within the biomedical sector. The objective was to investigate, critically discuss, and present the current knowledge and state-of-the-art practices, along with the challenges and perspectives of the most common applications. Through an extensive literature review, this work aims to supplement existing research, offering a comprehensive and up-to-date understanding of the subject. Biomedical research involving cryogenic mediums is advancing on multiple fronts, including the development of advanced medical technologies, clinical treatments for life-threatening conditions, high-quality biospecimen preservation, and antimicrobial interventions in industrial food processing. These advances open new horizons and present cutting-edge opportunities for research and the medical community. While the current body of evidence showcases the impressive impact of cryogenic mediums, such as nitrogen, helium, argon, and oxygen, on revolutionary developments, reaching definitive conclusions on their efficiency and safety remains challenging due to process complexity and research scarcity with a moderate certainty of evidence. Knowledge gaps further underline the need for additional studies to facilitate cryogenic research in developing innovative technological processes in biomedicine. These advancements have the potential to reshape the modern world and significantly enhance the quality of life for people worldwide.


Spatial neglect and perseveration in visuomotor exploration.
Right-brain-damaged patients with left spatial neglect make perseveration errors in target cancellation tasks. A recent study (Ronchi, Posteraro, Fortis, Bricolo, & Vallar, 2009) showed that perseveration occurred more frequently in star than in letter cancellation, possibly due to different stimulus disposition. In this study we investigated the role of the spatial organization of targets (scattered vs. arranged) of the star and letter cancellation tasks in eliciting omission and perseveration errors; the role of impairments of divided attentional resources; and the lesion correlates of perseveration. Thirty-three right-brain-damaged patients (27 with neglect, and six without neglect; 17 showing perseveration, 16 of them with spatial neglect) were given two versions of the star and letter cancellation tasks (with stimuli scattered or in rows), and a dual task. A scattered target disposition increased omission and perseveration errors. Target type modulated differently omission and perseveration, with the former being more elicited by verbal targets, and the latter occurring more frequently in the star cancellation task, with scattered stimuli. Perseveration behavior was unrelated to deficits of divided attention, as assessed by the dual task. Lesion analysis indicated damage to the right insula as a neural correlate of perseveration. A display including nonverbal (star) and scattered targets brings about more perseveration errors. Target type and organization modulate in a different fashion omission and perseveration, suggesting the involvement of independent pathological mechanisms, which, however, do not implicate deficits of divided attention. The role of insular damage in motor perseveration in spatial neglect is discussed.


Ultrasonography and color Doppler of proximal gluteal enthesitis in juvenile idiopathic arthritis: a descriptive study.
The presence of enthesitis (insertional inflammation) in patients with juvenile idiopathic arthritis (JIA) is difficult to establish clinically and may influence classification and treatment of the disease. We used ultrasonography (US) and color Doppler (CD) imaging to detect enthesitis at the small and deep-seated proximal insertion of the gluteus medius fascia on the posterior iliac crest where clinical diagnosis is difficult. The findings in JIA patients were compared with those obtained in healthy controls and with the patients' MRI results. Seventy-six proximal gluteus medius insertions were studied clinically (tenderness to palpation of the posterior iliac crest) and by US and CD (echogenicity, thickness, hyperemia) in 38 patients with JIA and in 38 healthy controls, respectively (median age 13 years, range 7-18 years). In addition, an additional MRI examination of the sacroiliac joints and iliac crests was performed in all patients. In patients with focal, palpable tenderness, US detected decreased echogenicity of the entheses in 53% of the iliac crests (bilateral in 37% and unilateral in 32%). US also revealed significantly thicker entheses in JIA patients compared to healthy controls (p < 0.003 left side, p < 0.001 right side). There was no significant difference in thickness between the left and right sides in individual subjects. Hyperemia was detected by CD in 37% (28/76) of the iliac crests and by contrast-enhanced MRI in 12% (6/50). According to US, the gluteus medius insertion was thicker in JIA patients than in controls, and it was hypoechoic (enthesitis) in about half of the patients. These findings may represent chronic, inactive disease in some of the patients, because there was only limited Doppler flow and MRI contrast enhancement. The present study indicates that US can be useful as an adjunct to clinical examination for improved assessment of enthesitis in JIA. This may influence disease classification, ambition to treat, and choice of treatment regimen.


The physiology of L-methionine catabolism to the secondary metabolite ethylene by Escherichia coli.
Catabolism of L-methionine by Escherichia coli strain B SPAO led to the formation of ethylene as a secondary metabolite (ethylenogenesis). Methionine was initially deaminated by a transamination reaction to the 2-oxo acid 2-oxo-4-methylthiobutyric acid (KMBA) which was then converted to ethylene. The utilization of L-methionine as an additional nitrogen source was investigated by examining ethylene synthesis under different nitrogen supply conditions. Ethylene formation in batch culture was unaffected by the concentration of the precursor L-methionine in the medium although increasing concentrations of NH4Cl resulted in progressively less ethylene formation. Cultures grown without L-methionine did not produce ethylene but were able to synthesize ethylene when L-methionine or KMBA was provided. Addition of L-tyrosine to batch cultures reduced the yield of ethylene after 42 h by 54%. Under these conditions the maximum transient level of KMBA was reduced by 32% and occurred later compared to when L-methionine was the only amino acid supplement. Continuous cultures grown under ammonia limitation produced both ethylene and KMBA. In contrast, when glucose was limiting, neither of these metabolites were produced. Cells harvested from continuous cultures grown under glucose or ammonia limitation were able to synthesize ethylene from either L-methionine or KMBA although their capacity for ethylene synthesis (ethylenogenic capacity) was optimal under ammonia limitation (C:N ratio = 20).


SDR-type human hydroxysteroid dehydrogenases involved in steroid hormone activation.
Hydroxysteroid dehydrogenases catalyze the NAD(P)(H)-dependent oxidoreduction of hydroxyl and oxo-functions at distinct positions of steroid hormones. This reversible reaction constitutes an important pre-receptor control mechanism for nuclear receptor ligands of the androgen, estrogen and glucocorticoid classes, since the conversion "switches" between receptor ligands and their inactive metabolites. The major reversible activities found in mammals acting on steroid hormones comprise 3alpha-, 11beta- and 17beta-hydroxysteroid dehydrogenases, and for each group several distinct isozymes have been described. The enzymes differ in their expression pattern, nucleotide cofactor preference, steroid substrate specificity and subcellular localization, and thus constitute a complex system ensuring cell-specific adaptation and regulation of steroid hormone levels. Several isoforms constitute promising drug targets, of particular importance in cancer, metabolic diseases, neurodegeneration and immunity.


Plaque quantification by coronary computed tomography angiography using intravascular ultrasound as a reference standard: a comparison between standard and last generation computed tomography scanners.
The emerging role of coronary computed tomography angiography (CCTA) as a non-invasive tool for atherosclerosis evaluation is supported by data reporting a good correlation between CCTA and intravascular ultrasound (IVUS) for plaque volume quantification. Aim of the present study was to evaluate whether a last generation CT-scanner may improve coronary plaque volume assessment using IVUS as standard-of-reference. From a registry of 1915 consecutive, all-comers, patients who underwent a clinically indicated IVUS evaluation we enrolled 59 patients who underwent CCTA with a 64-slice CT (Group 1) and 59 patients who underwent CCTA with whole-heart coverage CT scanner (Group 2). Patients who underwent CCTA with unfavourable heart rhythm were not excluded from the analysis. Image quality (4-point Likert scale) focused on plaque analysis was evaluated. Plaque volume quantification by CCTA was compared to IVUS. No difference in clinical characteristics was found between Group 1 and Group 2. Plaque volume quantification by CCTA was considered not feasible in 11 plaques of Group 1 and in 4 plaques of Group 2 (P = 0.09). Higher correlation for plaque volume quantification by CCTA vs. IVUS was demonstrated in Group 2 when compared with Group 1 (r = 0.9888 vs. 0.9499; P < 0.0001). The Bland-Altman analysis showed plaque volume overestimation by CCTA of 11.9 mm3 in Group 1 and 4 mm2 in Group 2 (P < 0.001). Effective radiation dose of CCTA was significantly lower in Group 2 vs. Group 1 (2.7 ± 0.9 vs. 8.1 ± 3.6 mSv, respectively; P < 0.001). CCTA using a new scanner generation showed to be an accurate non-invasive tool to assess and quantify coronary plaque volume.


[Significance of a mass survey for colorectal cancer at the workplace].
Two thousand four hundred and forty workers from 2 plants were enrolled in a mass survey for colorectal cancer using a single immunological fecal occult blood test as screening. The occult blood test was positive in 71 (4.0%) out of 1,759 subjects in whom the test was conducted. A further investigation, including total colonoscopy or barium enema, was performed on 52 (73.2%) of the 71. Colon cancer was detected in 5 cases: 1 with advanced cancer and 4 with early cancer (cancer in the adenoma). The advanced cancer was successfully resected by surgery and the early cancers were removed by endoscopic polypectomy. Adenomas were found in 21 cases, 11 of which were polypectomized. From these results, it can be seen that a survey for colorectal cancer at the workplace is important, and by expanding the number of subjects for the survey in the workplace we can expect more and earlier detections of colorectal cancer.


Do comorbidities predict pain and function in knee osteoarthritis following an exercise intervention, and do they moderate the effect of exercise? Analyses of data from three randomized controlled trials.
Although exercise is a core treatment for people with knee osteoarthritis (OA), it is currently unknown whether those with additional comorbidities respond differently to exercise than those without. We explored whether comorbidities predict pain and function following an exercise intervention in people with knee OA, and whether they moderate response to: exercise versus no exercise; and enhanced exercise versus usual exercise-based care. We undertook analyses of existing data from three randomized controlled trials (RCTs): TOPIK (n = 217), APEX (n = 352) and Benefits of Effective Exercise for knee Pain (BEEP) (n = 514). All three RCTs included: adults with knee pain attributable to OA; physiotherapy-led exercise; data on six comorbidities (overweight/obesity, pain elsewhere, anxiety/depression, cardiac problems, diabetes mellitus and respiratory conditions); the outcomes of interest (six-month Western Ontario and McMaster Universities Arthritis Index knee pain and function). Adjusted mixed models were fitted where data was available; otherwise linear regression models were used. Obesity compared with underweight/normal body mass index was significantly associated with knee pain following exercise, as was the presence compared with absence of anxiety/depression. The presence of cardiac problems was significantly associated with the effect of enhanced versus usual exercise-based care for knee function, indicating that enhanced exercise may be less effective for improving knee function in people with cardiac problems. Associations for all other potential prognostic factors and moderators were weak and not statistically significant. Obesity and anxiety/depression predicted pain and function outcomes in people offered an exercise intervention, but only the presence of cardiac problems might moderate the effect of exercise for knee OA. Further confirmatory investigations are required.


Binding of NO and CO to the d(1) Heme of cd(1) nitrite reductase from Pseudomonas aeruginosa.
The cd(1) nitrite reductase, a key enzyme in bacterial denitrification, catalyzes the one-electron reduction of nitrite to nitric oxide. The enzyme contains two redox centers, a c-type heme and a unique d(1) heme, which is a dioxoisobacteriochlorin. Nitric oxide, generated by this enzymatic pathway, if not removed from the medium, can bind to the ferrous d(1) cofactor with extremely high affinity and inhibit enzyme activity. In this paper, we report the resonance Raman investigation of the properties of nitric oxide and carbon monoxide binding to the d(1) site of the reduced enzyme. The Fe-ligand (Fe-NO and Fe-CO) stretching vibrational frequencies are unusually high in comparison to those of other ferrous heme complexes. The frequencies of the Fe-NO and N-O stretching modes appear at 585 and 1626 cm(-1), respectively, in the NO complex, while the frequencies of the Fe-CO and C-O stretching modes are at 563 and 1972 cm(-1), respectively, for the CO complex. Also, the widths (fwhm) of the Fe-CO and C-O stretching modes are smaller than those observed in the corresponding complexes of other heme proteins. The unusual spectroscopic characteristics of the d(1) cofactor are discussed in terms of both its unique electronic properties and the strongly polar distal environment around the iron-bound ligand. It is likely that the influence of a highly ruffled structure of heme d(1) on its electronic properties is the major factor causing anomalous Fe-ligand vibrational frequencies.


Pax-6 functions in boundary formation and axon guidance in the embryonic mouse forebrain.
The Pax-6 gene encodes a transcription factor that is expressed in regionally restricted patterns in the developing brain and eye. Here we describe Pax-6 expression in the early forebrain (prosencephalon) on embryonic day 9.5 (E9.5) to E10.5 using both whole-mount in situ hybridization and antibody labeling. We find close correlations between Pax-6+ domains and initial neural patterning, and identify corresponding defects in embryos homozygous for the Pax-6 allele, Small eye (Sey). Pax-6 expression defines the prosencephalon-mesencephalon boundary, and mutant embryos lack this morphological boundary. Markers of the caudal prosencephalon are lost (Pax-6, Lim-1, Gsh-1) and a marker for mesencephalon is expanded rostrally into the prosencephalon (Dbx). We conclude that the caudal prosencephalon (prosomere 1) is at least partially transformed to a mesencephalic fate. This transformation results in a specific deficit of posterior commissure axons. Sey/Sey embryos also exhibit an axon pathfinding defect specific to the first longitudinal tract in the prosencephalon (tpoc, tract of the postoptic commissure). In wild type, tpoc axons fan out upon coming in contact with a superficial patch of Pax-6+ neuron cell bodies. In the mutant, the tpoc axons have normal initial projections, but make dramatic errors where they contact the neuron cell bodies, and fail to pioneer this first tract. Thus Pax-6 is required for local navigational information used by axons passing through its domain of expression. We conclude that Pax-6 plays multiple roles in forebrain patterning, including boundary formation, regional patterning, neuron specification and axon guidance.


Targeted Proteomics to Assess the Response to Anti-Angiogenic Treatment in Human Glioblastoma (GBM).
Glioblastoma (GBM) is a highly aggressive primary brain tumor with dismal outcome for affected patients. Because of the significant neo-angiogenesis exhibited by GBMs, anti-angiogenic therapies have been intensively evaluated during the past years. Recent clinical studies were however disappointing, although a subpopulation of patients may benefit from such treatment. We have previously shown that anti-angiogenic targeting in GBM increases hypoxia and leads to a metabolic adaptation toward glycolysis, suggesting that combination treatments also targeting the glycolytic phenotype may be effective in GBM patients. The aim of this study was to identify marker proteins that are altered by treatment and may serve as a short term readout of anti-angiogenic therapy. Ultimately such proteins could be tested as markers of efficacy able to identify patient subpopulations responsive to the treatment. We applied a proteomics approach based on selected reaction monitoring (SRM) to precisely quantify targeted protein candidates, selected from pathways related to metabolism, apoptosis and angiogenesis. The workflow was developed in the context of patient-derived intracranial GBM xenografts developed in rodents and ensured the specific identification of human tumor versus rodent stroma-derived proteins. Quality control experiments were applied to assess sample heterogeneity and reproducibility of SRM assays at different levels. The data demonstrate that tumor specific proteins can be precisely quantified within complex biological samples, reliably identifying small concentration differences induced by the treatment. In line with previous work, we identified decreased levels of TCA cycle enzymes, including isocitrate dehydrogenase, whereas malectin, calnexin, and lactate dehydrogenase A were augmented after treatment. We propose the most responsive proteins of our subset as potential novel biomarkers to assess treatment response after anti-angiogenic therapy that warrant future analysis in clinical GBM samples.


Comparison of the effect of different methods of molt: production and welfare evaluation.
The purpose of this study was to evaluate alternative molting protocols assessing hen welfare and performance during and after molt. Hyline W-36 pullets were housed at 15 wk of age, and their egg production was obtained during this first cycle. When birds were 80 wk, the following molting treatments were applied: a conventional molt consisting of 10 d of fasting followed by cracked corn for 8 d and a pullet developer diet for 10 d; and 4 alternative molting programs: a soy hulls-based diet (12% CP, 1,455 kcal/kg of ME, and 1.38% Ca) offered for 14 d followed by cracked corn for 4 d and a pullet developer diet for 10 d; and the other 3 molt regimens consisted of feeding soy hulls for 4, 8, or 12 d followed by 10, 6, or 2 d, respectively, of a soy hulls-based diet and 4 d of cracked corn plus 10 d of a pullet developer diet. A nonmolted group of birds was fed a laying hen diet during the experimental period. Hen-day egg number was recorded daily for 56 wk (through 80-136 wk of age). The nonmolted hens showed lower hen-day production and fewer intact eggs and a higher number of cracked and shell-less eggs compared with those of the molted hens (P < 0.0001). A significant treatment by age effect (P < 0.0001) was observed for the variables of high-density lipoproteins and triglycerides. Control hens showed the lowest high-density lipoprotein concentration and the highest triglyceride levels at 84 wk of age when compared with all treatments. The lack of difference in heterophil-to-lymphocyte ratio during molt suggests little influence of molting protocols on this variable. Regardless of the treatments, molting was deleterious to bone quality. A high mobilization of Ca through bone resorption for eggshell formation could explain the results obtained. Behavioral patterns coincided with a decline in frustration activities and an increase in alertness as molt proceeded until 83 wk of age. Alternative molting diets consisting of soybean hulls were successful in providing acceptable postmolt egg production performance.


Alternative mitochondrial respiratory chains from two crustaceans: Artemia franciscana nauplii and the white shrimp, Litopenaeus vannamei.
Mitochondrial ATP is synthesized by coupling between the electron transport chain and complex V. In contrast, physiological uncoupling of these processes allows mitochondria to consume oxygen at high rates without ATP synthesis. Such uncoupling mechanisms prevent reactive oxygen species overproduction. One of these mechanisms are the alternative redox enzymes from the mitochondrial respiratory chain, which may help cells to maintain homeostasis under stress independently of ATP synthesis. To date, no reports have been published on alternative redox enzymes in crustaceans mitochondria. Specific inhibitors were used to identify alternative redox enzymes in mitochondria isolated from Artemia franciscana nauplii, and the white shrimp, Litopenaeus vannamei. We report the presence of two alternative redox enzymes in the respiratory chain of A. franciscana nauplii, whose isolated mitochondria used glycerol-3-phosphate as a substrate, suggesting the existence of a glycerol-3-phosphate dehydrogenase. In addition, cyanide and octyl-gallate were necessary to fully inhibit this species' mitochondrial oxygen consumption, suggesting an alternative oxidase is present. The in-gel activity analysis confirmed that additional mitochondrial redox proteins exist in A. franciscana. A mitochondrial glycerol-3-phosphate dehydrogenase oxidase was identified by protein sequencing as part of a branched respiratory chain, and an alternative oxidase was also identified in this species by western blot. These results indicate different adaptive mechanisms from artemia to face environmental challenges related to the changing levels of oxygen concentration in seawater through their life cycles. No alternative redox enzymes were found in shrimp mitochondria, further efforts will determine the existence of an uncoupling mechanism such as uncoupling proteins.


The fate of incomplete stent apposition with drug-eluting stents: an optical coherence tomography-based natural history study.
To assess the fate of incomplete stent apposition (ISA) after deployment of sirolimus-eluting stents (SESs). Thirty-two patients having intravascular ultrasound (IVUS)-guided PCI with SESs underwent assessment of stent deployment with quantitative coronary angiography, IVUS, and optical coherence tomography (OCT) pre-procedure, post-procedure, and at 10 months follow-up. Incomplete stent apposition was defined as separation of a stent strut from the inner vessel wall by >160 microm. At follow-up, 4.67% of struts with ISA at deployment failed to heal and 7.59% which were well apposed did not develop neointimal hyperplasia even after 10 months. Lesion remodelling was responsible for the development of late ISA in only 0.37% of struts. Failure of adequate neointimal hyperplasia was quantitatively the most important mechanism responsible for persistent acute ISA, classified in previous studies, which relied only on follow-up OCT, as late ISA. Thrombus was visualized in 20.6% of struts with ISA at follow-up and in 2.0% of struts with a good apposition (P < 0.001). In patients with SESs, ISA can fail to heal and even complete apposition can be associated with no neointimal hyperplasia. Incomplete stent apposition without neointimal hyperplasia was significantly associated with the presence of OCT-detected thrombus at follow-up, and may constitute a potent substrate for late stent thrombosis.


Reorganization in the human brain as illustrated by the thalamus.
A comparison is made between the relative size of the various thalamic nuclei in man and other primates. Using data for non-human primates predictions are made as to the expected size of the nuclei for the human brain. Of the nuclear groupings five are of the size predicted but three are not. The lateral geniculate is proportionately smaller than predicted, but it is argued that this need not imply a radical change.


Novel temporalis muscle and fat pad morphomic analyses aids preoperative risk evaluation and outcome assessment in nonsyndromic craniosynostosis.
Analytical morphomics is the term we created to describe an innovative, highly automated, anatomically indexed processing of 3D medical imaging data captured during the course of a patients' preoperative CT scan. Our specific aim is to determine the efficacy of craniofacial morphomic indices (CMI) such as temporalis muscle and temporal fat pad morphometric values to predict blood transfusion requirement and hospital stay in a cohort of children with nonsyndromic craniosynostosis (NSC). High-throughput, semi-automated image analysis was used to reconstruct the 3-dimensional anatomy of the temporalis muscle and temporal fat pad and to quantify CMIs. The prognostic effect of CMI on clinical outcomes were evaluated among all NSC patients and compared across various craniosynostosis subtypes using Wilcoxon nonparametric tests and Kendall's τ to determine significance. Using preoperative CT images, we evaluated 117 children with NSC from the University of Michigan Health System. Results demonstrate that increased temporal fat pad volume and local temporalis muscle volume are associated with better clinical outcomes in craniosynostosis patients. More specifically, temporal fat pad volume was shown to be a significant predictor of perioperative blood transfusion requirements (P = 0.0033) and increased temporal muscle volume correlated with decreased hospital stay (P = 0.016) when controlling for other covariates including age, sex, weight, and preoperative hematocrit. In addition, the same significant predictors were found when examining individual subtypes of craniosynostosis. Our findings demonstrate that maxillofacial CT scans provide a useful quantitative index reflecting general patient health, risk stratification, and probabilities of intervention in addition to their previously established ability to determine the specific pathology of the patient. We demonstrate that temporal morphomics predict the incidence of blood transfusion, hospital stay, and serve as a proxy for fitness in patients undergoing craniosynostosis surgery.


Gender differences in a randomized controlled trial treating tobacco use among adolescents and young adults with mental health concerns.
Treatment of tobacco use in mental health settings is rare despite high rates of comorbidity. With a focus on early intervention, we evaluated a tobacco treatment intervention among adolescents and young adults recruited from outpatient, school-based, and residential mental health settings and tested for gender differences. Intervention participants received computerized motivational feedback at baseline, 3 months, and 6 months and were offered 12 weeks of cessation counseling and nicotine patches. Usual care participants received a self-help guide and brief cessation advice. We examined 7-day point prevalence abstinence with biochemical confirmation at 3, 6, and 12 months; smoking reduction; and 24-hr quit attempts. At baseline, the sample (N = 60, 52% female, mean age = 19.5±2.9 years, 40% non-Hispanic Caucasian) averaged 7±6 cigarettes/day, 62% smoked daily, 38% smoked ≤ 30 min of waking, 12% intended to quit in the next month, 47% had a parent who smoked, and 3 of 5 of participants' closest friends smoked on average. During the 12-month study, 47% of the sample reduced their smoking, 80% quit for 24 hr, and 11%, 13%, and 17% confirmed 7-day point prevalence abstinence at 3-, 6-, and 12-month follow-up, respectively, with no differences by treatment condition (ps > .400). Over time, abstinence was greater among girls (adjusted odds ratio [AOR] = 8.9) than among boys, and abstinence was greater for lighter smokers than heavier smokers (AOR = 4.5) (p < .05). No mental health or other measured variables predicted abstinence. Adolescent and young adult smokers with mental health concerns are a challenging group to engage and to effectively treat for tobacco addiction, particularly heavier smokers and boys. Innovative approaches are needed.


Exercise is medicine in oncology: Engaging clinicians to help patients move through cancer.
Multiple organizations around the world have issued evidence-based exercise guidance for patients with cancer and cancer survivors. Recently, the American College of Sports Medicine has updated its exercise guidance for cancer prevention as well as for the prevention and treatment of a variety of cancer health-related outcomes (eg, fatigue, anxiety, depression, function, and quality of life). Despite these guidelines, the majority of people living with and beyond cancer are not regularly physically active. Among the reasons for this is a lack of clarity on the part of those who work in oncology clinical settings of their role in assessing, advising, and referring patients to exercise. The authors propose using the American College of Sports Medicine's Exercise Is Medicine initiative to address this practice gap. The simple proposal is for clinicians to assess, advise, and refer patients to either home-based or community-based exercise or for further evaluation and intervention in outpatient rehabilitation. To do this will require care coordination with appropriate professionals as well as change in the behaviors of clinicians, patients, and those who deliver the rehabilitation and exercise programming. Behavior change is one of many challenges to enacting the proposed practice changes. Other implementation challenges include capacity for triage and referral, the need for a program registry, costs and compensation, and workforce development. In conclusion, there is a call to action for key stakeholders to create the infrastructure and cultural adaptations needed so that all people living with and beyond cancer can be as active as is possible for them.


Prevalence and impact of occult hepatitis B infection in chronic hepatitis C patients treated with pegylated interferon and ribavirin.
The prevalence of occult hepatitis B, defined by absence of HBsAg and HBV DNA, ranges widely in patients with hepatitis C. This may influence the treatment of hepatitis C and the severity of liver disease. Sensitive and specific real-time PCR techniques are available commercially and can detect more reliably low HBV DNA levels. The aim of this study was to determine the prevalence of occult hepatitis B virus infection using the COBAS Taqman assay (Roche Diagnostics, Meylan, France) in the serum and liver of HBsAg negative patients with chronic hepatitis C and to evaluate its clinical consequences on liver pathology and its impact on the response to treatment with peg-IFNalpha and Ribavirin. HBV DNA detection was assessed retrospectively on 140 sera and 113 liver biopsies of HCV positive/HBsAg negative patients before treatment. A 4.4% (5/113) prevalence of occult hepatitis B was recorded in liver samples and in none of the sera. Anti-HBc was not detected in one, three of whom were sustained virological responders to treatment, one was relapsed responder and one was non-responder. Furthermore, in this cohort composed of 12% anti-HBs negative/anti-HBc positive and 20% anti-HBs positive/anti-HBc positive patients, anti-HBc was not associated with pre-therapeutic viral load, ALT serum levels, and histological activity or fibrosis. Using a commercial real-time PCR assay, we observed a low prevalence of occult B hepatitis. This, just as anti-HBC status, had no clinical impact in a large cohort of hepatitis C patients. It therefore does not appear useful to screen for occult hepatitis B in these patients with this test before beginning HCV treatment.


Protein kinases of malaria parasites: an update.
Protein kinases (PKs) play crucial roles in the control of proliferation and differentiation in eukaryotic cells. Research on protein phosphorylation has expanded tremendously in the past few years, in part as a consequence of the realization that PKs represent attractive drug targets in a variety of diseases. Activity in Plasmodium PK research has followed this trend, and several reports on various aspects of this subject were delivered at the Molecular Approaches to Malaria 2008 meeting (MAM2008), a sharp increase from the previous meeting. Here, the authors of most of these communications join to propose an integrated update of the development of the rapidly expanding field of Plasmodium kinomics.


Differential activation of alpha 1- and alpha 2-adrenoceptors on microvascular smooth muscle during sympathetic nerve stimulation.
The relative contribution of postjunctional alpha 1- and alpha 2-adrenoceptors to constriction of microvessels was examined during sympathetic nerve stimulation and sympathetic escape (difference between peak and steady-state constriction). Large arterioles (120 +/- 4 microns control diameter) and venules (174 +/- 6 microns) and small arterioles (13 +/- 4 microns) were examined in rat cremaster skeletal muscle during stimulation of the cremaster efferent innervation (decentralized lumbar sympathetic chain, 0.5-16 Hz, 2-minute train). The muscle was suspended in a tissue bath, and diameter was measured with intravital microscopy. Frequency-response curves were obtained after vehicle (prazosin or rauwolscine) was added to the bath. In large arterioles, prazosin (10(-7) M) significantly attenuated constriction by 60-80%; a fivefold higher concentration had no additional effect. In contrast, rauwolscine (1 to 5 x 10(-7) M) had no effect. Venules evidenced minimal response to nerve stimulation. In small arterioles, rauwolscine (5 x 10(-7) M) significantly attenuated constriction by 50-60%, while prazosin (10(-7) M) had no effect. These data suggest that for large arterioles, which are known to possess both receptors, alpha 1-adrenoceptors are preferentially stimulated by nerve-released norepinephrine. In contrast, sympathetic constriction of small arterioles is mediated by alpha 2-adrenoceptors. Compared with large arterioles, small arterioles exhibited greater peak and steady-state constriction at all frequencies, with maximal responses achieved over the 0.5-4 Hz range. Large arterioles exhibit graded constriction over the entire frequency range. Sympathetic escape exhibited a small, negatively correlated frequency dependence for large arterioles, tended to be greater for small arterioles, and was more evident in large arterioles during alpha 2-adrenoceptor constriction at low-frequency stimulation. This distinct neural control of large resistance vessels by alpha 1-adrenoceptors and small terminal arterioles by alpha 2-adrenoceptors may allow neurogenic regulation of these vessel segments to be differentially susceptible to modulation by other extrinsic and intrinsic vasoactive controls that preferentially interact with alpha 1- and alpha 2-adrenergic contractile mechanisms.


Conceptualising engagement with HIV care for people on treatment: the Indicators of HIV Care and AntiRetroviral Engagement (InCARE) Framework.
As the crisis-based approach to HIV care evolves to chronic disease management, supporting ongoing engagement with HIV care is increasingly important to achieve long-term treatment success. However, 'engagement' is a complex concept and ambiguous definitions limit its evaluation. To guide engagement evaluation and development of interventions to improve HIV outcomes, we sought to identify critical, measurable dimensions of engagement with HIV care for people on treatment from a health service-delivery perspective. We used a pragmatic, iterative approach to develop a framework, combining insights from researcher experience, a narrative literature review, framework mapping, expert stakeholder input and a formal scoping review of engagement measures. These inputs helped to refine the inclusion and definition of important elements of engagement behaviour that could be evaluated by the health system. The final framework presents engagement with HIV care as a dynamic behaviour that people practice rather than an individual characteristic or permanent state, so that people can be variably engaged at different points in their treatment journey. Engagement with HIV care for those on treatment is represented by three measurable dimensions: 'retention' (interaction with health services), 'adherence' (pill-taking behaviour), and 'active self-management' (ownership and self-management of care). Engagement is the product of wider contextual, health system and personal factors, and engagement in all dimensions facilitates successful treatment outcomes, such as virologic suppression and good health. While retention and adherence together may lead to treatment success at a particular point, this framework hypothesises that active self-management sustains treatment success over time. Thus, evaluation of all three core dimensions is crucial to realise the individual, societal and public health benefits of antiretroviral treatment programmes. This framework distils a complex concept into three core, measurable dimensions critical for the maintenance of engagement. It characterises elements that the system might assess to evaluate engagement more comprehensively at individual and programmatic levels, and suggests that active self-management is an important consideration to support lifelong optimal engagement. This framework could be helpful in practice to guide the development of more nuanced interventions that improve long-term treatment success and help maintain momentum in controlling a changing epidemic.


Visual scanning behaviours and their role in the navigation of the Australian desert ant Melophorus bagoti.
Ants are excellent navigators, using a combination of innate strategies and learnt information to guide habitual routes. The mechanisms underlying this behaviour are little understood though one avenue of investigation is to explore how innate sensori-motor routines are used to accomplish route navigation. For instance, Australian desert ant foragers are occasionally observed to cease translation and rotate on the spot. Here, we investigate this behaviour using high-speed videography and computational analysis. We find that scanning behaviour is saccadic with pauses separated by fast rotations. Further, we have identified four situations where scanning is typically displayed: (1) by naïve ants on their first departure from the nest; (2) by experienced ants departing from the nest for their first foraging trip of the day; (3) by experienced ants when the familiar visual surround was experimentally modified, in which case frequency and duration of scans were proportional to the degree of modification; (4) when the information from visual cues is at odds with the direction indicated by the ant's path integration system. Taken together, we see a general relationship between scanning behaviours and periods of uncertainty.


Phase I study of lovastatin, an inhibitor of the mevalonate pathway, in patients with cancer.
Lovastatin, an inhibitor of the enzyme 3-hydroxy-3-methylglutaryl-coenzyme A reductase (the major regulatory enzyme of the mevalonate pathway of cholesterol synthesis), displays antitumor activity in experimental models. We therefore conducted a Phase I trial to characterize the tolerability of lovastatin administered at progressively higher doses to cancer patients. From January 1992 to July 1994, 88 patients with solid tumors (median age, 57 +/- 14 years) were treated p.o. with 7-day courses of lovastatin given monthly at doses ranging from 2 to 45 mg/kg/day. The inhibitory effects of lovastatin were monitored through serum concentrations of cholesterol and ubiquinone, two end products of the mevalonate pathway. Concentrations of lovastatin and its active metabolites were also determined, by bioassay, in the serum of selected patients. Cyclical treatment with lovastatin markedly inhibited the mevalonate pathway, evidenced by reductions in both cholesterol and ubiquinone concentrations, by up to 43 and 49% of pretreatment values, respectively. The effect was transient, however, and its magnitude appeared to be dose independent. Drug concentrations reached up to 3.9 micrometer and were in the range associated with antiproliferative activity in vitro. Myopathy was the dose-limiting toxicity. Other toxicities included nausea, diarrhea, and fatigue. Treatment with ubiquinone was associated with reversal of lovastatin-induced myopathy, and its prophylactic administration prevented the development of this toxicity in a cohort of 56 patients. One minor response was documented in a patient with recurrent high-grade glioma. Lovastatin given p.o. at a dose of 25 mg/kg daily for 7 consecutive days is well tolerated. The occurrence of myopathy, the dose-limiting toxicity, can be prevented by ubiquinone supplementation. To improve on the transient inhibitory activity of this dosing regimen on the mevalonate pathway, alternative schedules based on uninterrupted administration of lovastatin should also be studied.


The development of a radioimmunoassay procedure for the estimation of Tamm-Horsfall glycoprotein in human serum.
1. A quantitative radioimmunoassay was developed for the measurement of nanogram amounts of Tamm-Horsfall (TH) glycoprotein in the presence of serum proteins at concentrations above 30 mg/ml. 2. Specific anti-(TH-glycoprotein) antibodies were labelled with 125I and these were usable for a period of 8 weeks. 3. Agarose beads (Sepharose 4B), to which TH-glycoprotein had been coupled via cyanogen bromide activation of the Sepharose, were used as the solid phase in the assay. This proved stable upon storage at 4 degrees C for periods in excess of 4 months. 4. The dissociation of the glycoprotein in the presence of serum proteins that was necessary for quantification was achieved by subjecting the sample to ultrasonication. 5. Assays conducted on a small sample of normal serum produced evidence that normal serum contained amounts (50-180 ng/ml) of a substance that reacted similarly to TH-glycoprotein in the assay procedure and in a series of experiments conducted to confirm the presence of this substance in human serum.


Phosphorylation by casein kinase 2 facilitates rRNA gene transcription by promoting dissociation of TIF-IA from elongating RNA polymerase I.
The protein kinase casein kinase 2 (CK2) phosphorylates different components of the RNA polymerase I (Pol I) transcription machinery and exerts a positive effect on rRNA gene (rDNA) transcription. Here we show that CK2 phosphorylates the transcription initiation factor TIF-IA at serines 170 and 172 (Ser170/172), and this phosphorylation triggers the release of TIF-IA from Pol I after transcription initiation. Inhibition of Ser170/172 phosphorylation or covalent tethering of TIF-IA to the RPA43 subunit of Pol I inhibits rDNA transcription, leading to perturbation of nucleolar structure and cell cycle arrest. Fluorescence recovery after photobleaching and chromatin immunoprecipitation experiments demonstrate that dissociation of TIF-IA from Pol I is a prerequisite for proper transcription elongation. In support of phosphorylation of TIF-IA switching from the initiation into the elongation phase, dephosphorylation of Ser170/172 by FCP1 facilitates the reassociation of TIF-IA with Pol I, allowing a new round of rDNA transcription. The results reveal a mechanism by which the functional interplay between CK2 and FCP1 sustains multiple rounds of Pol I transcription.


A graph-based approach to diploid genome assembly.
Constructing high-quality haplotype-resolved de novo assemblies of diploid genomes is important for revealing the full extent of structural variation and its role in health and disease. Current assembly approaches often collapse the two sequences into one haploid consensus sequence and, therefore, fail to capture the diploid nature of the organism under study. Thus, building an assembler capable of producing accurate and complete diploid assemblies, while being resource-efficient with respect to sequencing costs, is a key challenge to be addressed by the bioinformatics community. We present a novel graph-based approach to diploid assembly, which combines accurate Illumina data and long-read Pacific Biosciences (PacBio) data. We demonstrate the effectiveness of our method on a pseudo-diploid yeast genome and show that we require as little as 50× coverage Illumina data and 10× PacBio data to generate accurate and complete assemblies. Additionally, we show that our approach has the ability to detect and phase structural variants. https://github.com/whatshap/whatshap. Supplementary data are available at Bioinformatics online.


TP53 mutations and MDM2(SNP309) identify subgroups of AML patients with impaired outcome.
TP53 is commonly mutated in several cancers and confers treatment resistance and poor prognosis. Altered expression of mouse double minute 2 (MDM2), a negative regulator of p53, may also attenuate normal p53 signaling, thereby enhancing tumor transformation and resistance to apoptosis. The single nucleotide polymorphism (SNP) 309 has been reported to increase MDM2 expression and impair normal p53 response. We investigated the frequency and impact of TP53 mutations (TP53mut) and MDM2(SNP309) on treatment outcome and overall survival (OS) in 189 Swedish acute myeloid leukemia patients. The genetic analyses were performed using SSCA and direct sequencing (for mutations in exon 5-8 of TP53) and Pyrosequencing (for the MDM2(SNP309) ). We found a high frequency (22%) of TP53mut in patients with cytogenetic aberrations, with association to high-risk cytogenetics (P < 0.001). TP53mut patients had lower response rates (22% compared with 76% CR in TP53 wild-type (wt) patients, P < 0.001) and reduced OS (2 and 16 months, respectively, P < 0.001). In TP53wt patients with high or intermediate risk cytogenetic aberrations, the MDM2(SNP309) conferred an impaired outcome, with patients carrying the alternative G-allele having shorter OS compared with T/T patients (median 9 vs. 50 months, P = 0.020). Our results show that TP53mut analysis and MDM2(SNP309) genotyping may be useful tools for prognostication, risk stratification, and selection of patients most likely to benefit from new drugs targeting the p53 signaling pathway.


Trajectory shifts in the Arctic and subarctic freshwater cycle.
Manifold changes in the freshwater cycle of high-latitude lands and oceans have been reported in the past few years. A synthesis of these changes in freshwater sources and in ocean freshwater storage illustrates the complementary and synoptic temporal pattern and magnitude of these changes over the past 50 years. Increasing river discharge anomalies and excess net precipitation on the ocean contributed approximately 20,000 cubic kilometers of fresh water to the Arctic and high-latitude North Atlantic oceans from lows in the 1960s to highs in the 1990s. Sea ice attrition provided another approximately 15,000 cubic kilometers, and glacial melt added approximately 2000 cubic kilometers. The sum of anomalous inputs from these freshwater sources matched the amount and rate at which fresh water accumulated in the North Atlantic during much of the period from 1965 through 1995. The changes in freshwater inputs and ocean storage occurred in conjunction with the amplifying North Atlantic Oscillation and rising air temperatures. Fresh water may now be accumulating in the Arctic Ocean and will likely be exported southward if and when the North Atlantic Oscillation enters into a new high phase.


Polycaprolactone fibres as a potential delivery system for collagen to support bone regeneration.
Poly(ε-caprolactone) (PCL) is biocompatible, non-immunogenic and non-toxic, and slowly degrades, allowing sufficient time for tissue regeneration. PCL has the potential for application in bone and cartilage repair as it may provide the essential structure required for bone regeneration, however, an ideal scaffold system is still undeveloped. PCL fibres were prepared using the gravity spinning technique, in which collagen was either incorporated into or coated onto the 'as-spun' fibres, in order to develop novel biodegradable polymer fibres which will effectively deliver collagen and support the attachment and proliferation of human osteoblast (HOB) cells for bone regeneration. The physical and mechanical characteristics and cell fibre interactions were analysed. The PCL fibres were found to be highly flexible and inclusion of collagen did not alter the mechanical properties of PCL fibres. Overall, HOB cells were shown to effectively adhere and proliferate on all fibre platforms tested, although proliferation rates were enhanced by surface coating PCL fibres with collagen compared to PCL fibres incorporating collagen and PCL-only fibres. These findings highlight the potential of using gravity spun PCL fibres as a delivery platform for extracellular matrix proteins, such as collagen, in order to enhance cell adherence and proliferation for tissue repair.


Charge carrier-selective contacts for nanowire solar cells.
Charge carrier-selective contacts transform a light-absorbing semiconductor into a photovoltaic device. Current record efficiency solar cells nearly all use advanced heterojunction contacts that simultaneously provide carrier selectivity and contact passivation. One remaining challenge with heterojunction contacts is the tradeoff between better carrier selectivity/contact passivation (thicker layers) and better carrier extraction (thinner layers). Here we demonstrate that the nanowire geometry can remove this tradeoff by utilizing a permanent local gate (molybdenum oxide surface layer) to control the carrier selectivity of an adjacent ohmic metal contact. We show an open-circuit voltage increase for single indium phosphide nanowire solar cells by up to 335 mV, ultimately reaching 835 mV, and a reduction in open-circuit voltage spread from 303 to 105 mV after application of the surface gate. Importantly, reference experiments show that the carriers are not extracted via the molybdenum oxide but the ohmic metal contacts at the wire ends.


Diagnosing early cognitive decline-When, how and for whom?
Mild cognitive impairment (MCI) is a term used to describe cognitive impairment in one or more cognitive domains that is greater than any expected age-related changes, but not of the magnitude to warrant a diagnosis of dementia. This review considers how early cognitive decline is diagnosed, focusing on the use of neuropsychological tests and neuroimaging, as well as the differential diagnosis. Potential treatments, including secondary prevention, post-diagnostic support and self-help are discussed. Finally, medico-legal matters such as driving, lasting power of attorney and employment are outlined.


Correlation Between Apolipoprotein M and Inflammatory Factors in Obese Patients.
BACKGROUND The aim of this study was to observe apolipoprotein M (ApoM) level in obese patients and to explore its correlation with inflammatory factors. MATERIAL AND METHODS A total number of 96 participants were recruited and divided into 2 groups: the control group (or healthy group) whose participants had normal body weight (n=58), and the obese group with all its participants diagnosed with obesity (n=38). Data on blood pressure, body weight, height, body mass index, diastolic function of brachial artery endothelium, fasting venous blood glucose, blood lipids, plasmatic ApoM, interleukin-6 (IL-6), C-reactive protein (CRP), tumor necrosis factor-α (TNF-α), fasting insulin, and adiponectin levels were collected for both groups. RESULTS In the obese group, the levels of plasmatic ApoM, high-density lipoprotein cholesterol (HDL-C), and plasmatic adiponectin were significantly (p<0.05) decreased compared to the control group, and the levels of IL-6, TNF-α, CRP, and fasting insulin were significantly increased (p<0.05) compared to the control group. For the obese group, plasmatic ApoM level was positively correlated with HDL-C level and negatively correlated with levels of IL-6, TNF-α, CRP, insulin, and insulin resistance index. However, no significant correlations were revealed between plasmatic ApoM and the diastolic function of brachial artery endothelium, adiponectin level, blood pressure, and blood glucose level. CONCLUSIONS Obese patients showed significantly lower plasmatic ApoM levels than people with normal body weight, and ApoM level showed a strong correlation with CRP, TNF-α, and IL-6 levels, which indicated that ApoM might be regulated by these inflammatory factors.


Micro-integrated extended cavity diode lasers for precision potassium spectroscopy in space.
We present a micro-integrated, extended cavity diode laser module for space-based experiments on potassium Bose-Einstein condensates and atom interferometry. The module emits at the wavelength of the potassium D2-line at 766.7 nm and provides 27.5 GHz of continuous tunability. It features sub-100 kHz short term (100 μs) emission linewidth. To qualify the extended cavity diode laser module for quantum optics experiments in space, vibration tests (8.1 g(RMS) and 21.4 g(RMS)) and mechanical shock tests (1500 g) were carried out. No degradation of the electro-optical performance was observed.


Castration-Resistant Prostate Cancer: AUA Guideline Amendment 2015.
The purpose of this amendment is to incorporate relevant newly-published literature to better provide a rational basis for the management of patients with castration-resistant prostate cancer. The original systematic review and meta-analysis of the published literature yielded 303 studies published from 1996 through 2013. This review informed the majority of the guideline statements. Clinical Principles and Expert Opinions were used for guideline statements lacking sufficient evidence. In April 2014, the CRPC guideline underwent amendment based on an additional literature search, which retrieved additional studies published between February 2013 and February 2014. Thirty-seven studies from this search provided data relevant to the specific treatment modalities for CRPC. In March 2015, the CRPC guideline underwent a second amendment, which incorporated 10 additional studies into the evidence base published through February 2015. Guideline statements based on six index patients developed to represent the most common scenarios encountered in clinical practice were amended appropriately. The additional literature provided the basis for an update of current supporting text as well as the incorporation of new guideline statements for multiple index patients. Given the rapidly evolving nature of this field, this guideline should be used in conjunction with recent systematic literature reviews and an understanding of the individual patient's treatment goals. Patients' preferences and personal goals should be considered when choosing management strategies. This guideline will be continually updated as new literature emerges in the field.


Examining trust factors in online food risk information: The case of unpasteurized or 'raw' milk.
The internet has become an increasingly important way of communicating with consumers about food risk information. However, relatively little is known about how consumers evaluate and come to trust the information they encounter online. Using the example of unpasteurized or raw milk this paper presents two studies exploring the trust factors associated with online information about the risks and benefits of raw milk consumption. In the first study, eye-tracking data was collected from 33 pasteurised milk consumers whilst they viewed six different milk related websites. A descriptive analysis of the eye-tracking data was conducted to explore viewing patterns. Reports revealed the importance of images as a way of capturing initial attention and foregrounding other features and highlighted the significance of introductory text within a homepage. In the second, qualitative study, 41 consumers, some of whom drank raw milk, viewed a selection of milk related websites before participating in either a group discussion or interview. Seventeen of the participants also took part in a follow up telephone interview 2 weeks later. The qualitative data supports the importance of good design whilst noting that balance, authorship agenda, the nature of evidence and personal relevance were also key factors affecting consumers trust judgements. The results of both studies provide support for a staged approach to online trust in which consumers engage in a more rapid, heuristic assessment of a site before moving on to a more in-depth evaluation of the information available. Findings are discussed in relation to the development of trustworthy online food safety resources.


Self-driven filter-based blood plasma separator microfluidic chip for point-of-care testing.
There is currently a growing need for lab-on-a-chip devices for use in clinical analysis and diagnostics, especially in the area of patient care. The first step in most blood assays is plasma extraction from whole blood. This paper presents a novel, self-driven blood plasma separation microfluidic chip, which can extract more than 0.1 μl plasma from a single droplet of undiluted fresh human blood (~5 μl). This volume of blood plasma is extracted from whole blood with high purity (more than 98%) in a reasonable time frame (3 to 5 min), and without the need for any external force. This would be the first step towards the realization of a single-use, self-blood test that does not require any external force or power source to deliver and analyze a fresh whole-blood sample, in contrast to the existing time-consuming conventional blood analysis. The prototypes are manufactured in polydimethylsiloxane that has been modified with a strong nonionic surfactant (Silwet L-77) to achieve hydrophilic behavior. The main advantage of this microfluidic chip design is the clogging delay in the filtration area, which results in an increased amount of extracted plasma (0.1 μl). Moreover, the plasma can be collected in one or more 10 μm-deep channels to facilitate the detection and readout of multiple blood assays. This high volume of extracted plasma is achieved thanks to a novel design that combines maximum pumping efficiency without disturbing the red blood cells' trajectory through the use of different hydrodynamic principles, such as a constriction effect and a symmetrical filtration mode. To demonstrate the microfluidic chip's functionality, we designed and fabricated a novel hybrid microdevice that exhibits the benefits of both microfluidics and lateral flow immunochromatographic tests. The performance of the presented hybrid microdevice is validated using rapid detection of thyroid stimulating hormone within a single droplet of whole blood.


Toward structural femtosecond chemical dynamics: imaging chemistry in space and time.
We aim to observe a chemical reaction in real time using gas-phase X-ray diffraction. In our initial experiment at the Linac Coherent Light Source (LCLS), we investigated the model system 1,3-cyclohexadiene (CHD) at very low vapor pressures. This reaction serves as a benchmark for numerous transformations in organic synthesis and natural product biology. Excitation of CHD by an ultraviolet optical pulse initiates an electrocyclic reaction that transforms the closed ring system into the open-chain structure of 1,3,5-hexatriene. We describe technical points of the experimental method and present first results. We also outline an approach to analyze the data involving nonlinear least-square optimization routines that match the experimental observations with predicted diffraction patterns calculated from trajectories for nonadiabatic vibronic wave packets.


Effects of a series of 4-alkyl analogues of 3,5-diethoxycarbonyl-1,4-dihydro-2,4,6-trimethylpyridine on the major inducible cytochrome P-450 isozymes of rat liver.
Various 4-alkyl analogues of 3,5-diethoxycarbonyl-1,4-dihydro-2,4,6-trimethylpyridine (DDC) cause mechanism-based inactivation of cytochrome P-450 (P-450) by destroying the heme prosthetic group. We have examined the isozyme selectivity of representative DDC analogues with respect to the major inducible P-450 isozymes of rat liver. Hepatic microsomes from untreated, phenobarbital (PB)-treated, beta-naphthoflavone (beta NF)-treated, and dexamethasone (DEX)-treated rats were incubated with a DDC analogue and NADPH and were subsequently analyzed for P-450 and heme content, P-450 isozyme immunoreactivity, and enzyme activity. Compared with the uninduced state, 4-isopropyl-DDC caused slightly less P-450 destruction following beta NF induction and much greater destruction following DEX pretreatment. Also, 4-hexyl-DDC was found to cause less P-450 destruction following PB or DEX pretreatment, compared with results obtained with untreated rats. These results suggest that DDC analogues possess different isozyme selectivity profiles. Monoclonal antibodies (MAbs) directed against the major inducible isozymes of P-450 were used to probe Western blots of microsomal protein following DDC analogue treatment. The formation of lower molecular mass (45-55 kDa) immunoreactive proteins in microsomes from beta NF-treated rats following DDC analogue treatment was revealed by two MAbs (1-31-2 and 1-36-1), suggesting that the apoprotein of the major beta NF-inducible isozyme, P-450c, is subject to alteration by DDC analogues. In microsomes from DEX-treated rats, DDC analogues caused the formation of higher molecular mass (80, 94, and 115 kDa) proteins showing immunoreactivity with MAb 2-13-1, directed against a major DEX-inducible isozyme belonging to the P-450p family. These immunochemical findings are supported by the demonstration that DDC analogues also caused mechanism-based inhibition of the catalytic activity of P-450c (7-ethoxyresorufin O-deethylase) and P-450p (erythromycin N-demethylase) but not that of the major PB-inducible isozyme, P-450b (7-pentoxyresorufin O-dealkylase). The combined immunochemical and enzymic studies indicate that rat liver P-450 c and p are targets for mechanism-based inactivation by DDC analogues.


A serial study of erythrocyte sodium content and sodium pump kinetics in pregnancy.
1. Normotensive primigravid pregnant women were studied longitudinally during pregnancy and 20 weeks after delivery. 2. Erythrocyte sodium content, ouabain-sensitive sodium flux and sodium pump rate constant were measured in whole blood, and the maximum velocity and sodium affinity of the sodium pump were measured in vitro. 3. Erythrocyte sodium content decreased and the sodium pump rate constant increased up to 26 weeks gestation. The increase in rate constant was due to an increase in the affinity of the sodium pump for sodium up to 20 weeks gestation. After 20 weeks gestation there was an increase in maximum velocity and a decrease in sodium affinity of the sodium pump but no further change in the sodium pump rate constant. 4. At 14 weeks gestation the sodium pump rate constant was correlated with both the maximum velocity and sodium affinity constant. After this time the relationship was much more variable and there was no correlation with the sodium affinity constant. The comparison of measurements of the sodium pump in whole blood and in vitro gave no evidence of sodium pump inhibition. 5. The erythrocyte sodium pump changed throughout gestation with different components to the change, but, overall, available sodium pump activity in blood increased and sodium content decreased.


Social bees are fitter in more biodiverse environments.
Bee population declines are often linked to human impacts, especially habitat and biodiversity loss, but empirical evidence is lacking. To clarify the link between biodiversity loss and bee decline, we examined how floral diversity affects (reproductive) fitness and population growth of a social stingless bee. For the first time, we related available resource diversity and abundance to resource (quality and quantity) intake and colony reproduction, over more than two years. Our results reveal plant diversity as key driver of bee fitness. Social bee colonies were fitter and their populations grew faster in more florally diverse environments due to a continuous supply of food resources. Colonies responded to high plant diversity with increased resource intake and colony food stores. Our findings thus point to biodiversity loss as main reason for the observed bee decline.


[Smoking and chronic obstructive pulmonary disease: attributable risk determination].
Smoking is the main risk factor for Chronic Obstructive Pulmonary Disease (COPD), an important cause of morbidity and mortality. To estimate smoking attributable risk and population attributable risk in COPD patients attended in Public Health Services of Santiago. A case control study matched by sex and age was carried out. Crude and adjusted attributable risks as well as population attributable risk were estimated, controlled by potential confounders and by interaction variables. Mean ages for cases and controls were 68 and 67 years respectively. When compared to the control group, COPD patients had a higher smoking prevalence (at least 100 cigarettes in their life span: 89.7% vs 60.3%; p <0.01). Among COPD patients, heavy smokers proportion was 4 times higher than in controls, they smoked for more years (43 vs 31; p <0.01) and more cigarettes per day (18 vs 5; p <0.01). Adjusted attributable risk was 87% (95% Confidence Interval (CI): 63.7-94.8). If a patient smoked at least 100 cigarettes in his/her life span and this risk was 92.7% (CI: 82.4-96.9) for heavy smokers. Projecting this index to Santiago inhabitants, about 87,000 individuals older than 40 years would be suffering COPD due to smoking. This article confirms the strong association between smoking and COPD. Attributable risks are high and significant, even when they are adjusted by confounding variables. Women had a higher risk than men, at lower levels of tobacco consumption.


Changes in CRH and ACTH synthesis during experimental and human septic shock.
The mechanisms of septic shock-associated adrenal insufficiency remain unclear. This study aimed at investigating the synthesis of corticotropin-releasing hormone (CRH) and vasopressin (AVP) by parvocellular neurons and the antehypophyseal expression of ACTH in human septic shock and in an experimental model of sepsis. To test the hypothesis that ACTH secretion is decreased secondarily to alteration of CRH or AVP synthesis, we undertook a neuropathological study of the antehypophyseal system in patients who had died from septic shock and rats with experimental faecal peritonitis. Brains obtained in 9 septic shock patients were compared to 10 nonseptic patients (controls). Parvocellular expression of AVP and CRH mRNA were evaluated by in situ hybridization. Antehypophyseal expression of ACTH, vasopressin V1b and CRH R1 receptors and parvocellular expression of iNOS in the PVN were evaluated by immunohistochemistry. The same experiments were carried out in a fecal peritonitis-induced model of sepsis. Data from septic rats with (n = 6) or without (n = 10) early death were compared to sham-operated (n = 8) animals. In patients and rats, septic shock was associated with a decreased expression of ACTH, unchanged expression of V1B receptor, CRHR1 and AVP mRNA, and increased expression of parvocellular iNOS compared to controls. Septic shock was also characterized by an increased expression of CRH mRNA in rats but not in patients, who notably had a greater duration of septic shock. The present study suggests that in humans and in rats, septic shock is associated with decreased ACTH synthesis that is not compensated by its two natural secretagogues, AVP and CRH. One underlying mechanism might be increased expression of iNOS in hypothalamic parvocellular neurons.