Contribution of different anatomical and physiologic factors to iris contour and anterior chamber angle changes during pupil dilation: theoretical analysis.
To investigate the contribution of three anatomical and physiologic factors (dilator thickness, dynamic pupillary block, and iris compressibility) to changes in iris configuration and anterior chamber angle during pupil dilation. A MATHEMATICAL MODEL OF THE ANTERIOR SEGMENT BASED ON THE AVERAGE VALUES OF OCULAR DIMENSIONS WAS DEVELOPED TO SIMULATE PUPIL DILATION. TO CHANGE THE PUPIL DIAMETER FROM 3.0 TO 5.4 MM IN 10 SECONDS, ACTIVE DILATOR CONTRACTION WAS APPLIED BY IMPOSING STRESS IN THE DILATOR REGION. THREE SETS OF PARAMETERS WERE VARIED IN THE SIMULATIONS: (1) a thin (4 μm, 1% of full thickness) versus a thick dilator (covering the full thickness iris) to quantify the effects of dilator anatomy, (2) in the presence (+PB) versus absence of pupillary block (-PB) to quantify the effect of dynamic motion of aqueous humor from the posterior to the anterior chamber, and (3) a compressible versus an incompressible iris to quantify the effects of iris volume change. Changes in the apparent iris-lens contact and angle open distance (AOD500) were calculated for each case. The thin case predicted a significant increase (average 700%) in iris curvature compared with the thick case (average 70%), showing that the anatomy of dilator plays an important role in iris deformation during dilation. In the presence of pupillary block (+PB), AOD500 decreased 25% and 36% for the compressible and incompressible iris, respectively. Iris bowing during dilation was driven primarily by posterior location of the dilator muscle and by dynamic pupillary block, but the effect of pupillary block was not as large as that of the dilator anatomy according to the quantified values of AOD500. Incompressibility of the iris, in contrast, had a relatively small effect on iris curvature but a large effect on AOD500; thus, we conclude that all three effects are important.


Application of STS markers for leaf rust resistance genes in near-isogenic lines of spring wheat cv. Thatcher.
Sequence tagged site (STS) markers for eight resistance genes against Puccinia recondita f. sp. tritici were used to screen a set of near-isogenic lines of wheat cv. Thatcher containing in total 40 different Lr genes and their alleles. Polymerase chain reaction (PCR) analysis was carried out by using STS, SCAR and CAPS primers specific for the leaf rust resistance genes Lr1, Lr9, Lr10, Lr19, Lr24, Lr28, Lr37 and Lr47. The STS, CAPS and SCAR markers linked to resistance genes Lr9, Lr10, Lr19, Lr24, Lr37 and Lr47 were found to be reliable in diverse genetic backgrounds. The amplification product of the Lr1 gene marker was detected in the susceptible cv. Thatcher and in all of the near-isogenic lines examined except Lr2a, Lr2b, Lr2c and Lr19. The sequence analysis of PCR products amplified in lines Lr1, Lr10, Lr28 and in cv. Thatcher indicated that the near-isogenic lines and cv. Thatcher contained in the targeted chromosome region an allele that differed from the original alleles corresponding to Lr1/6*Thatcher (TLR621) and susceptible Thatcher (TH621). The amplification product specific to the STS marker of the Lr1 gene was amplified in almost all Thatcher near-isogenic lines and in cv. Thatcher because their alleles possessed primer sequences identical to the original allele TLR621. The marker for the Lr28 resistance gene was identified in line Lr28, carrying gene Lr28, and in 21 other near-isogenic lines. The sequencing of PCR products specific to Lr28 and generated in lines Lr1, Lr10 and Lr28 indicated that the lines Lr1, Lr10 and Lr28 are heterozygous in this region.


Association between arterial stiffness and peritoneal small solute transport rate.
While cardiovascular disease accounts for 40-50% of the mortality in dialysis patients, and while a high peritoneal transport in continuous ambulatory peritoneal dialysis (CAPD) is an independent predictor of outcome, it is unclear if there are any links. Aortic stiffness has become established as a cardiovascular risk factor. We thus studied pulse wave velocity (PWV) in CAPD patients to explore the possible link between peritoneal small solute transport and aortic stiffness. CAPD patients (n = 76, 27 M/49 F) in our center were included in the present study. Aortic stiffness was assessed by brachial pulse pressure (PP) and carotid-femoral PWV. Patients' peritoneal small solute transport rate was assessed by D/P(cr) at 4 h. Extracellular water over total body water (E/T ratio) was assessed by means of bioimpedance analysis. C-reactive protein was also measured. Carotid-femoral PWV was positively associated with patients' age (r = 0.555; P < 0.01), time on peritoneal dialysis (r = 0.332; P < 0.01), diabetic status (r = 0.319; P < 0.01), D/P(cr) (r = 0.241; P < 0.05), PP (r = 0.475; P < 0.01), and E/T (r = 0.606; P < 0.01). In a multivariate regression analysis, carotid-femoral PWV was independently determined by E/T (P < 0.01), PP (P < 0.01), age (P < 0.01), and D/P(cr) (P < 0.05). D/P(cr), in addition to E/T, age, and PP, was an independent predictor of elevated carotid-femoral PWV in CAPD patients, suggesting that there might be a link between high aortic stiffness and increased peritoneal small solute transport rate.


Adolescents' expectancies for smoking to regulate affect predict smoking behavior and nicotine dependence over time.
Mounting evidence suggests that individuals smoke, in part, to regulate affective experience (e.g., tension reduction, mood enhancement). Implicit in such motives is the expectancy or belief that smoking will decrease negative affect and increase positive affect. The contribution of cognitively-driven expectancies to the initiation and continuation of smoking during adolescence remains largely uninvestigated. The current study examined the influence of negative affect relief expectancies (NAREs) for smoking on smoking behavior and nicotine dependence using longitudinal data from a study on the emotional and social contexts of youth smoking. Participants were 568 adolescents with smoking experience (mean age 15.67, 56.7% female). Three separate mixed regression models were estimated to determine the relative contribution of NAREs to smoking behavior and nicotine dependence measured at 4 time points over 2 years. NAREs for smoking influenced all smoking outcomes at baseline and predicted increases in smoking behavior and nicotine dependence over time, even after controlling for anxious and depressive symptoms and baseline nicotine dependence. Outcome expectancies for affect management emerged as an important risk factor for smoking escalation and the development of nicotine dependence during adolescence. The present findings highlight the potential importance of cognitively-driven expectancies as a risk factor for smoking escalation during this critical developmental period.


'Intern life': a longitudinal study of burnout, empathy, and coping strategies used by French GPs in training.
More than half of French medical GP trainees (GPTs) suffer from burnout. To define and follow the evolution of risk factors, such as empathy and coping strategies, associated with burnout in this population. Prospective longitudinal study involving volunteers of 577 Parisian university GPTs in 2012. Self-reported anonymous online questionnaires were sent three times every 6 months to all participants. Stress was measured using the Intern-Life scale and burnout using the Maslach Inventory, and anxiety and depression measured using the Hospital Anxiety and Depression Scale (HADS). Sociodemographic, professional, and personal data, including coping strategies and measures of empathy were also collected. In total 343 questionnaires were fully completed at baseline (T0): 304 were usable at baseline, 169 were usable at 6 months (T1) and 174 at 1 year (T2). Stress rates decreased sharply between T1 (scores 42.96) and T2 (17.08), while scores for burnout remained relatively stable: more than 13% of GPTs had high scores in all three dimensions of burnout. Depersonalisation increased from 61% (T1) to 66% (T2). One hundred and four paired samples were analysed between T0 and T1, and between T1 and T2. Emotion-centred coping was associated with emotional exhaustion (P<0.05), while professional support reduced it. Experiences of aggression increased depersonalisation (P<0.05). Social support, problem-centred coping, perspective-taking empathy, and professional support improved the sense of personal accomplishment (P<0.05). Tools to help GPTs are available but are underused. More training in doctor-patient relationships and understanding of medical hidden curricula are necessary to decrease burnout among GPTs and improve their wellbeing and patient care.


Assessment of GFR by four methods in adults in Ashanti, Ghana: the need for an eGFR equation for lean African populations.
Equations for estimating glomerular filtration rate (GFR) have not been validated in Sub-Saharan African populations, and data on GFR are few. GFR by creatinine clearance (Ccr) using 24-hour urine collections and estimated GFR (eGFR) using the four-variable Modification of Diet in Renal Disease (MDRD-4)[creatinine calibrated to isotope dilution mass spectrometry (IDMS) standard], Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) and Cockcroft-Gault equations were obtained in Ghanaians aged 40-75. The population comprised 1013 inhabitants in 12 villages; 944 provided a serum creatinine and two 24-hour urines. The mean weight was 54.4 kg; mean body mass index was 21.1 kg/m(2). Mean GFR by Ccr was 84.1 ml/min/1.73 m(2); 86.8% of participants had a GFR of >/=60 ml/min/1.73 m(2). Mean MDRD-4 eGFR was 102.3 ml/min/1.73 m(2) (difference vs. Ccr, 18.2: 95% CI: 16.8-19.5); when the factor for black race was omitted, the value (mean 84.6 ml/min/1.73 m(2)) was close to Ccr. Mean CKD-EPI eGFR was 103.1 ml/min/1.73 m(2), and 89.4 ml/min/1.73 m(2) when the factor for race was omitted. The Cockcroft-Gault equation underestimated GFR compared with Ccr by 9.4 ml/min/1.73 m(2) (CI: 8.3-10.6); particularly in older age groups. GFR by Ccr, and eGFR by MDRD-4, CKD-EPI and Cockcroft-Gault showed falls with age: MDRD-4 5.5, Ccr 7.7, CKD-EPI 8.8 and Cockcroft-Gault 11.0 ml/min/1.73 m(2)/10 years. The percentage of individuals identified with CKD stages 3-5 depended on the method used: MDRD-4 1.6% (7.2 % without factor for black race; CKD-EPI 1.7% (4.7% without factor for black race), Ccr 13.2% and Cockcroft-Gault 21.0%. Mean eGFR by both MDRD-4 and CKD-EPI was considerably higher than GFR by Ccr and Cockcroft-Gault, a difference that may be attributable to leanness. MDRD-4 appeared to underestimate the fall in GFR with age compared with the three other measurements; the fall with CKD-EPI without the adjustment for race was the closest to that of Ccr. An equation tailored specifically to the needs of the lean populations of Africa is urgently needed. For the present, the CKD-EPI equation without the adjustment for black race appears to be the most useful.


Diabetes, depressive symptoms, and functional disability in African Americans: the Jackson Heart Study.
To investigate the degree to which comorbid depression contributes to the relationship of diabetes with functional disability in African Americans (AAs), a population at high-risk for complications. We examined 2989 African Americans (AAs) in the Jackson Heart Study who had diabetes and depressive symptoms (CES-D) assessed at baseline. Overall functional disability was defined as the inability to perform at least one task of daily living. Multivariable logistic regression models explored the association of diabetes and depressive symptoms with functional disability. Prevalence of overall functional disability was highest with both diabetes and depressive symptoms (54%), similar with diabetes alone (31%) or depressive symptoms alone (33%), and lowest with neither (15%). Adjusting for demographics, smoking, BMI, cardiovascular comorbidities, and hsCRP, the association of depressive symptoms alone (OR=2.30,95% CI 1.75-3.03) and both diabetes and depressive symptoms (OR=2.75,1.88-4.04) with overall functional disability was significant, but not for diabetes alone (OR=1.26,0.95-1.67), compared to neither. In regression analyses including any diabetes and any depressive symptoms together in models, the main effect of depressive symptoms but not diabetes was associated with overall functional disability, and the interaction term was not significant (p-value=0.84). Functional disability was highest among AAs who have both diabetes and depressive symptoms; the latter was a stronger contributor. Future studies should explore mechanisms underlying functional disability in diabetes, particularly the role of depression.


Function of the MAPK scaffold protein, Ste5, requires a cryptic PH domain.
Ste5, the prototypic mitogen-activated protein kinase (MAPK) scaffold protein, associates with plasma membrane-tethered Gbetagamma freed upon pheromone receptor occupancy, thereby initiating downstream signaling. We demonstrate that this interaction and membrane binding of an N-terminal amphipathic alpha-helix (PM motif) are not sufficient for Ste5 action. Rather, Ste5 contains a pleckstrin-homology (PH) domain (residues 388-518) that is essential for its membrane recruitment and function. Altering residues (R407S K411S) equivalent to those that mediate phosphoinositide binding in other PH domains abolishes Ste5 function. The isolated PH domain, but not a R407S K411S derivative, binds phosphoinositides in vitro. Ste5(R407S K411S) is expressed normally, retains Gbetagamma and Ste11 binding, and oligomerizes, yet is not recruited to the membrane in response to pheromone. Artificial membrane tethering of Ste5(R407S K411S) restores signaling. R407S K411S loss-of-function mutations abrogate the constitutive activity of gain-of-function Ste5 alleles, including one (P44L) that increases membrane affinity of the PM motif. Thus, the PH domain is essential for stable membrane recruitment of Ste5, and this association is critical for initiation of downstream signaling because it allows Ste5-bound Ste11 (MAPKKK) to be activated by membrane-bound Ste20 (MAPKKKK).


Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Infectivity by Viral Load, S Gene Variants and Demographic Factors, and the Utility of Lateral Flow Devices to Prevent Transmission.
How severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infectivity varies with viral load is incompletely understood. Whether rapid point-of-care antigen lateral flow devices (LFDs) detect most potential transmission sources despite imperfect clinical sensitivity is unknown. We combined SARS-CoV-2 testing and contact tracing data from England between 1 September 2020 and 28 February 2021. We used multivariable logistic regression to investigate relationships between polymerase chain reaction (PCR)-confirmed infection in contacts of community-diagnosed cases and index case viral load, S gene target failure (proxy for B.1.1.7 infection), demographics, SARS-CoV-2 incidence, social deprivation, and contact event type. We used LFD performance to simulate the proportion of cases with a PCR-positive contact expected to be detected using 1 of 4 LFDs. In total, 231 498/2 474 066 (9%) contacts of 1 064 004 index cases tested PCR-positive. PCR-positive results in contacts independently increased with higher case viral loads (lower cycle threshold [Ct] values), for example, 11.7% (95% confidence interval [CI] 11.5-12.0%) at Ct = 15 and 4.5% (95% CI 4.4-4.6%) at Ct = 30. B.1.1.7 infection increased PCR-positive results by ~50%, (eg, 1.55-fold, 95% CI 1.49-1.61, at Ct = 20). PCR-positive results were most common in household contacts (at Ct = 20.1, 8.7% [95% CI 8.6-8.9%]), followed by household visitors (7.1% [95% CI 6.8-7.3%]), contacts at events/activities (5.2% [95% CI 4.9-5.4%]), work/education (4.6% [95% CI 4.4-4.8%]), and least common after outdoor contact (2.9% [95% CI 2.3-3.8%]). Contacts of children were the least likely to test positive, particularly following contact outdoors or at work/education. The most and least sensitive LFDs would detect 89.5% (95% CI 89.4-89.6%) and 83.0% (95% CI 82.8-83.1%) of cases with PCR-positive contacts, respectively. SARS-CoV-2 infectivity varies by case viral load, contact event type, and age. Those with high viral loads are the most infectious. B.1.1.7 increased transmission by ~50%. The best performing LFDs detect most infectious cases.


Effects of reservoir system historical evolution on water system landscape patterns: the case of Changshou, China.
Reservoirs are a crucial form of water ecological infrastructure that offers a wide range of ecosystem services. The impact of reservoir construction on land use and landscape patterns is a matter of considerable importance. However, the specific effects of local-scale reservoir development on water system landscape patterns have not been comprehensively investigated. The Changshou district of Chongqing, located in southwest China and having an artificial lake system primarily composed of reservoirs, is an excellent example of examining the effects of human activities on the landscape patterns of water systems. This research reconstructed the historical evolution of reservoirs and rivers in Changshou District by historical maps and remote sensing data over seven decades from 1950 to 2020. On this basis, the landscape pattern metrics and graph-theoretical metrics of water systems in four crucial years, 1950, 1960, 1990, and 2020, were analyzed. The results indicate that from 1950 to 2020, the reservoir area in Changshou District increased by 64.195 km2, while the river area decreased by 7.297 km2; the two periods with the highest reservoir construction intensity were the 1950s and the 1970s to 1980s, when the reservoir area increased by 58.456 km2 and 3.089 km2, respectively; the construction of reservoirs diminished the connectivity of water systems and increased their fragmentation, dispersion, and edge complexity, but had little effect on their concentration and shape complexity. The findings can be applied to the optimization of water system patterns at the local level, the management of reservoir groups, the decision-making of water ecological infrastructure, and ecological effect studies of artificial lakes.


Modulation of Kv3 potassium channels expressed in CHO cells by a nitric oxide-activated phosphatase.
1.Voltage-gated K+ channels containing Kv3 subunits play specific roles in the repolarization of action potentials. Kv3 channels are expressed in selective populations of CNS neurons and are thought to be important in facilitating sustained and/or repetitive high frequency firing. Regulation of the activity of Kv3 channels by neurotransmitters could have profound effects on the repetitive firing characteristics of those neurons. 2.Kv3 channels are found in several neuronal populations in the CNS that express nitric oxide synthases (NOSs). We therefore investigated whether Kv3 channels are modulated by the signalling gas nitric oxide (NO). 3. We found that Kv3.1 and Kv3.2 currents are potentially suppressed by D-NONOate and other NO donors. The effects of NO on these currents are mediated by the activation of guanylyl cyclase (GC), since they are prevented by Methylene Blue, an inhibitor of GC, and by ODQ, a specific inhibitor of the soluble form of GC. Moreover, application of 8-Br-cGMP, a permeant analogue of cGMP, also blocked Kv3.1 and Kv3.2 currents. 4.KT5283, a cGMP-dependent protein kinase (PKG) blocker, prevented the inhibition of Kv3.1 and Kv3.2 currents by D-NONOate and 8-Br-cGMP. This indicates that activation of PKG as a result of the increase in intracellular cGMP levels produced by D-NONOate or 8-Br-cGMP is necessary for channel block. 5. Although the effects of NO on Kv3.1 and Kv3.2 channels require PKG activity, two observations suggest that they are not mediated by phosphorylation of channel proteins: (a) the reagents affect both Kv3.2 and Kv3.1 channels, although only Kv3.2 proteins have a putative PKA-PKG phosphorylation site, and (b) mutation of the PKA-PKG phosphorylation site in Kv3.2 does not interfere with the effects of NO or cGMP. 6. The inhibitory effects of NO and cGMP on Kv3.1 and Kv3.2 currents appear to be mediated by the activation of serine-threonine phosphatase, since they are blocked by low doses of okadaic acid. Furthermore, direct intracellular application of the catalytic subunit of protein phosphatase 2A inhibited Kv3.2 currents, indicating that activity of PKG-induced phosphatase is necessary and sufficient to inhibit these channels. 7. The results suggest that basal phosphorylation of Kv3 channel proteins is required for proper channel function. Activation of phosphatases via NO or other signals that increase cGMP might be a potent mechanism to regulate Kv3 channel activity in neurons.


Ex situ cultivation protocol for Cystoseira amentacea var. stricta (Fucales, Phaeophyceae) from a restoration perspective.
Due to multiple impacts, Cystoseira forests are experiencing a significant decline, which is affecting the ecosystem services they provide. Despite conservation efforts, there is an urgent need to develop best practices and large-scale restoration strategies. To implement restoration actions, we developed an ex situ protocol for the cultivation of Cystoseira. amentacea var. stricta, aimed at reducing the time needed for laboratory culture, thus avoiding prolonged maintenance and minimizing costs. Specifically, we tested the effects of temperature, light and substratum on settlement and growth of early life stages using a factorial experiment. Temperature (20 and 24°C) and photoperiod (15L:9D) were selected to reflect the conditions experienced in the field during the reproductive period. Two light intensities (125 and 250 μmol photons m-2s-1) were selected to mimic the condition experienced in the absence of canopy (i.e. barren-higher light intensity) or in the understory (lower light intensity) during gamete release. The tested substrata were flat polished pebbles and rough clay tiles. The release of gametes and the successive survival and development of embryo and germlings were followed for two weeks. Regardless of the culture conditions, rougher tiles showed higher zygote settlement, but the substrata did not affect the successive development. Zygote mortality after one week averaged 50% and at the end of the second week, embryonic survival was higher under lower light and temperature conditions, which also determined the growth of larger embryos.


Weight loss with a modified Mediterranean-type diet using fat modification: a randomized controlled trial.
There is evidence that Mediterranean diets with a high proportion of olive oil and nuts can be effective for weight management and prevention of cardiovascular disease. It might be difficult for populations with other eating habits to follow such diets. Therefore, a modified Mediterranean-type diet using fat modification through neutral and butter-flavored canola oil, walnuts and walnut oil with two portion-controlled sweet daily snacks was tested in Germany. Randomized waiting-list control study with overweight/grade 1 obese subjects: 12-week self-help modified Mediterranean-type diet, 6 weeks of diet plans and 6 weeks of weight loss maintenance training. Trial duration was 12 months. Intervention group (IG) included 100 participants (average age of 52.4 years, weight 85.1 kg and body mass index (BMI) 30.1 kg/m(2)), waiting-list control group (CG) included 112 participants (52.6 years, 84.1 kg and 30.1 kg/m(2)). Per-protocol weight loss after 12 weeks was 5.2 kg in IG vs 0.4 kg in CG (P ⩽ 0.0001), BMI -1.8 vs -0.1 kg/m(2) (P ⩽ 0.0001), waist circumference -4.7 vs -0.9 cm (P ⩽ 0.0001). Triglycerides, total cholesterol and LDL cholesterol improved significantly in IG but not in CG. One-year dropouts: 44% in IG and 53% in CG. Weight loss after 12 months: 4.2 kg (pooled data). A five-meal modified Mediterranean-type diet with two daily portion-controlled sweet snacks was effective for weight management in a self-help setting for overweight and grade 1 obese subjects. Fat modification through canola oil, walnuts and walnut oil improved blood lipids even at 12 months.


Long-term follow-up study of unruptured vertebral artery dissection: clinical outcomes and serial angiographic findings.
Although the spontaneous occurrence of an unruptured vertebral artery (VA) dissection has increasingly been recognized as a relatively common cause of stroke, and the clinical aspects of this lesion have gradually been determined, its natural course remains obscure. The main goal of this study was to clarify the management protocol for this condition by examining serial angiographic changes in patients with unruptured VA dissections. Seventeen patients with unruptured VA dissections, including 13 men and four women, were clinically and angiographically examined between 1993 and 1998. All patients were observed using serial angiography studies. The initial angiography examinations most frequently revealed stenotic lesions (appearance of a pearl-and-string sign or string sign) in eight (47.1%) of 17 cases. In 15 cases (88.2%), changes in the lesions were evident on follow-up angiography studies. Stenotic lesions resulted in occlusion in four cases, normalization in three, and subsequent formation of an aneurysm in one case, which was treated successfully by proximal occlusion of the affected vessel performed using a detachable balloon. Occluded lesions, which were initially observed in three patients, recanalized in two patients and remained unchanged in one patient. Fusiform dilation alone was demonstrated in three patients during the initial angiography session; these lesions became normalized or were unchanged on follow-up studies. Saccular aneurysms were observed in two patients. In one of these cases, proximal ligation of the parent artery was successfully performed because of subsequent aneurysm enlargement. A double lumen, which appeared in one patient with an extradural VA dissection, became occluded. Magnetic resonance T2-weighted imaging studies revealed infarction corresponding to the posterior circulation in seven cases. During long-term observation in this series, good or excellent recovery was obtained in 14 (87.5%) of 16 patients, and moderate or severe disability in two (12.5%); one patient was lost to follow up after the second angiography study. A follow-up angiography study must be performed during the early stage (within approximately 3 weeks after onset of symptoms) to confirm the formation or enlargement of an aneurysm, because such conditions may be amenable to surgical treatment. Unruptured VA dissection could otherwise be treated and followed conservatively. Although the majority of dissected lesions seem likely to stabilize within a few months, as evidenced on angiography, in some cases a longer observation period is required.


Identification of novel therapeutic targets for contrast induced acute kidney injury (CI-AKI): alpha blockers as a therapeutic strategy for CI-AKI.
Iodinated contrast is used for imaging and invasive procedures and it can cause contrast induced acute kidney injury (CI-AKI), which is the third leading hospital-acquired health problem. The purpose of the present study was to determine the effect of α-adrenergic receptor-1b (Adra1b) inhibition by using terazosin on change in kidney function, gene, and protein expression in C57BL/6J male mice, 6-8 weeks with chronic kidney disease (CKD). CKD was induced by surgical nephrectomy. Twenty eight days later, 100-µL of iodinated contrast (CI group) or saline (S group) was given via the carotid artery. Whole-transcriptome RNA-sequencing (RNA-Seq) analysis of the kidneys was performed at day 2. Mice received either 50-µL of saline ip or terazosin (2 mg/kg) in 50-µL of saline ip 1 hour before contrast administration which was continued every 12 hours until the animals were euthanized 2 and 7 days later. The kidneys were removed for gene expression, immunohistochemical analysis, and blood serum analyzed for kidney function. Differential gene expression analysis identified 21 upregulated and 436 downregulated genes (fold change >2; P < 0.05) that were common to all sample (n = 3 for both contrast and saline). We identified Adra1b using bioinformatic analysis. Mice treated with terazosin had a significant decrease in serum creatinine, urinary Kim-1 levels, HIF-1α, apoptosis, and downstream Adrab1 genes including Ece1, Edn1, pMAPK14 with increased cell proliferation. Contrast exposure upregulated Adra1b gene expression in HK-2 cells. Inhibition of Adra1b with terazosin abrogated Ece1, Edn1, and contrast-induced Fsp-1, Mmp-2, Mmp-9 expression, and caspase-3/7 activity in HK-2 cells.


Infant apnea: polygraphic studies and follow-up monitoring.
Twenty-six infants with unexplained and apparently life-threatening apnea were evaluated clinically, underwent eight- or 12-hour polygraphic recordings during sleep, and were then observed on home monitors. The one exception was an infant who was treated with theophylline and not monitored at home. Clinical evaluation revealed a history of vomiting in 19 infants, gastroesophageal reflux (GER) on esophagram in 19, and subtle neurologic abnormalities in ten. Polygraphic studies revealed questionable EEG abnormalities in nine infants, abnormal rapid eye movement (REM) time in four, prolonged apnea in one, and increased frequency of brief mixed (central and obstructive) and obstructive apneas in five. Of the 11 infants who underwent pH monitoring during their polygraphic sleep studies, seven had at least one recorded episode of GER, but the episodes were not accompanied by apnea or bradycardia. Thirteen infants had a subsequent episode of apnea that required stimulation, and in eight infants, cardiopulmonary resuscitation was given. There was two deaths. Two infants subsequently developed seizure disorders. Three of the five infants who underwent surgical fundoplication had recurrent apneic episodes. GER commonly occurred in these infant apnea patients but did not correlate with the clinical outcome. Although subtle abnormalities may be detected by sophisticated polygraphic monitoring studies, they are not predictive of recurrent apnea or death. Home monitoring may be useful in detecting the infant at risk for recurrent life-threatening apnea and in providing some safeguard for that infant.


Statins and diabetes.
Lipid abnormalities play an important part in raising the cardiovascular risk in diabetic subjects. The main components of diabetic dyslipidemia are increased plasma triglycerides, low concentration of high-density lipoprotein cholesterol, preponderance of small, dense low-density lipoprotein, and excessive postprandial lipemia. Small, dense low-density lipoprotein, the elevation in remnant triglyceride-rich lipoprotein particles, and the low high-density lipoprotein are the most powerful atherogenic components. The coexistence of these three factors strongly aggravates the lipid accumulation in the arterial wall and the formation of atherosclerotic plaques. The position of diabetes in cardiovascular risk assessment has been recently reviewed in the Harmonized Clinical Guidelines on Prevention of Atherosclerotic Vascular Disease. In general, patients with diabetes carry a high risk for cardiovascular disease, but the absolute risk varies depending on the type of diabetes, age, and population baseline risk. The Adult Treatment Program III (ATP III) and the American Heart Association have designated diabetes as a high-risk condition and recommended intensive risk-factor management. Concerning therapeutic targets, both ATP III and the American Diabetes Association (ADA) guidelines have identified low-density lipoprotein cholesterol as the first priority of lipid lowering, and the optimal level was set at less than 2.6 mmol/L (100 mg/dL). There is strong evidence, coming from landmark secondary prevention studies, that LDL lowering in people with diabetes is associated with significant clinical benefits. The benefits of statin therapy in type 2 diabetics can no longer be questioned. Ongoing clinical trials will help clarify the question of whether increasing high-density lipoprotein cholesterol with fibrates in the presence of low low-density lipoprotein levels (lower than 3.4 mmol/L, or 130 mg/dL) will be more beneficial than statin therapy alone. The new paradigms in risk-reduction therapies for type 2 diabetic subjects are focused on cardiovascular disease prevention, rather than only on glucose or lipid control. Therapeutic lifestyle changes are considered primary therapies for hyperglycemia and coexisting metabolic syndrome, which can be diagnosed in more than half of type 2 diabetes subjects. New perspectives of lipid management in type 2 diabetes should take into account that insulin resistance, increased lipolysis, and overproduction of large, buoyant, very low density lipoprotein particles are at the base of diabetic dyslipidemia. Accordingly, drugs acting in the regulatory steps of very low density lipoprotein assembly should be developed. Activation of peroxisome proliferator activated receptor alpha (PPARalpha), as occurs with fibrates, lowers free fatty acids (FFAs) and triglyceride levels. PPARgamma agonism, as demonstrated by the thiazolidinediones, increases triglyceride lipolysis, FFA transport, and conversion of FFAs to triglycerides. As separate activation of PPARalpha and PPARgamma improves lipid metabolism, the development of new drugs integrating PPARalpha and PPARgamma activity (PPAR-alpha/gamma agonists) is a promising line that may further improve insulin resistance, FFA metabolism, and consequently, atherogenic diabetic dyslipidemia.


Minimum 5-Year Outcomes After Primary Segmental Labral Reconstruction for Irreparable Labral Tears in the Hip With Hamstring Grafts: With a Subanalysis Comparing Autograft Versus Allograft.
Comparable short-term outcomes have been obtained using hamstring allografts versus autografts after primary segmental labral reconstruction (SLR). Midterm results have not yet been determined. (1) To evaluate minimum 5-year patient-reported outcome (PRO) scores in patients who underwent primary SLR with hamstring grafts in the setting of femoroacetabular impingement syndrome (FAIS) and irreparable labral tears and (2) to compare the outcomes of hamstring autografts versus allografts in a subanalysis using propensity-matched groups. Cohort study; Level of evidence, 3. Prospectively collected data were retrospectively reviewed for patients who underwent primary hip arthroscopy between September 2010 and November 2015. Patients were included if they underwent SLR using hamstring autografts or allografts and had preoperative and minimum 5-year PROs. The exclusion criteria were previous ipsilateral hip surgery or conditions, dysplasia, or Tönnis grade >1. Patients with autograft SLR were propensity matched 1 to 1 based on age, sex, and body mass index (BMI) to patients who underwent SLR using hamstring allografts. The minimal clinically important difference (MCID) and the Patient Acceptable Symptom State (PASS) were calculated. Overall, 48 patients (N = hips 48) were eligible to be included in this study, and 41 patients (n = 41 hips [85.4%]) had a minimum 5-year follow-up reporting significant improvements in all PROs. Within the entire cohort, 9.8% required a secondary arthroscopy, with a mean time of 19 ± 1.8 months, and survivorship was 82.9%. Of the 41 included patients, 15 underwent an SLR with a hamstring autograft and were matched to 15 patients with labral reconstruction using a hamstring allograft. Groups were similar for sex (P > .999), age (P = .775), and BMI (P = .486). The mean follow-up times were 80.8 ± 25.5 and 66.1 ± 8.3 months (P = .223) for the autograft and allograft groups, respectively. Baseline PROs, preoperative radiographic measurements, surgical findings, and intraoperative procedures were similar. The groups achieved significant and comparable improvements for all PROs (P < .0001), satisfaction (P = .187), and the rate of achieving the MCID and the PASS. However, a tendency for higher postoperative PROs favoring allograft reconstruction was found. At a minimum 5-year follow-up, patients who underwent primary arthroscopic SLR in the context of FAIS and irreparable labra, with either autograft or allograft hamstring tendons, reported significant improvements and comparable postoperative scores for all PROs, patient satisfaction, MCID, and PASS.


Plasma requirement for expression of lupus-like anticoagulant.
Described here are five patients with lupus-like anticoagulants, four of whom required coincubation of normal plasma in order to inhibit the pro-coagulant activity of crude brain phospholipid. It is suggested that this plasma requirement for expression of anticoagulant activity is similar or identical to the "lupus-cofactor" effect described by earlier observers.


Chromoblastomycosis with Squamous Cell Carcinoma.
A 47 year old male patient had extensive verrucous lesions on the right upper limb for 33 years. One of the lesions-4eveloped ulceration which had not healed for6 years. A clinical diagnosis of chromoblastomycosis was confirmed histopathologically and mycologically, Cladosporium cladosporioides being the organism cultured. Combined treatment with amphotericin B and 5fluorouracil gave good response with most lesions resolving. A few verrucous plaques and the ulcer persisted. Biopsy from the ulcer revealed a low - grade squamous cell carcinoma which was treated with excision and grafting. A year later, the patient developed metastasis in the axillary lymph nodes which also showed fungal granuloma at the time of biopsy. The axilla was irradiated with 4500 rads and three courses of bleomycin injections were given. However, he died 10 months later following a massive bleed from an axillary vessel.


Interleukin-17A antagonist attenuates radiation-induced lung injuries in mice.
To investigate the effect of interleukin-17A (IL-17A) antibodies on radiation-induced lung injuries in mice. The thorax of 135 mice were divided into Sham (n = 30), radiation control (RC, n = 35), treatment (n = 35, IL-17A-neutralizing antibody, 4 μg/mouse, IV, 4 days per month for 4 months) and placebo group (n = 35) before a single dose irradiation (15 Gy) to the thorax. Inflammation and collagen contents in the lung tissues were examined, and the concentration of IL-17A, TGF-β1, and IL-6 in bronchoalveolar lavage fluid (BALF) were measured. In another 50 animals, 180-day survival rate following the irradiation and treatment was calculated by Kaplan-Meier method. Sixteen weeks after the irradiation and treatment, there was significant inflammatory cell infiltration and interstitial collagen depositions in the radiation control and placebo groups, whereas these changes were relatively mild in the treatment group. The percentage of grade II and III alveolitis in the treatment group (16%, P < .05) was lower than in the RC (72%) or placebo group (64%). The mean Aschcroft fibrosis scores were 2.8 (treatment group), 5.2 (RC), and 4.8 (placebo group), respectively. The scores of treatment group was lower than that of RC (P < .001) or placebo group (P < .001). The IL-17A, TGF-β, and IL-6 concentrations in the treatment group were lower than in the RC and placebo group (P < .01) following the irradiation. The 180-day mortality rate in the treatment group was lower than in the RC group 16.7% versus 75.0%, P < .05). IL-17A antibody treatment alleviates radiation-induced pneumonitis and subsequent fibrosis, and improvise postirradiation survival.


The Role of the Insulin-Like Growth Factor 1 Pathway in Immune Tumor Microenvironment and Its Clinical Ramifications in Gynecologic Malignancies.
Treatment of patients with gynecologic malignancies diagnosed at advanced stages remains a therapeutic challenge. Survival rates of these patients remain significantly low, despite surgery and chemotherapy. Advances in understanding the role of the immune system in the pathogenesis of cancer have led to the rapid evolution of immunotherapeutic approaches. Immunotherapeutic strategies, including targeting specific immune checkpoints, as well as dendritic cell (DC) immunotherapy are being investigated in several malignancies, including gynecological cancers. Another important approach in cancer therapy is to inhibit molecular pathways that are crucial for tumor growth and maintenance, such as the insulin-like growth factor-1 (IGF1) pathway. The IGF axis has been shown to play a significant role in carcinogenesis of several types of tissue, including ovarian cancer. Preclinical studies reported significant anti-proliferative activity of IGF1 receptor (IGF1R) inhibitors in gynecologic malignancies. However, recent clinical studies have shown variable response rates with advanced solid tumors. This study provides an overview on current immunotherapy strategies and on IGF-targeted therapy for gynecologic malignancies. We focus on the involvement of IGF1R signaling in DCs and present our preliminary results which imply that the IGF axis contributes to an immunosuppressive tumor microenvironment (TME). For the long term, we believe that restoring the TME function by IGF1R targeting in combination with immunotherapy can serve as a new clinical approach for gynecological cancers.


20p12.3 microdeletion predisposes to Wolff-Parkinson-White syndrome with variable neurocognitive deficits.
Wolff-Parkinson-White syndrome (WPW) is a bypass re-entrant tachycardia that results from an abnormal connection between the atria and ventricles. Mutations in PRKAG2 have been described in patients with familial WPW syndrome and hypertrophic cardiomyopathy. Based on the role of bone morphogenetic protein (BMP) signalling in the development of annulus fibrosus in mice, it has been proposed that BMP signalling through the type 1a receptor and other downstream components may play a role in pre-excitation. Using the array comparative genomic hybridisation (CGH), we identified five individuals with non-recurrent deletions of 20p12.3. Four of these individuals had WPW syndrome with variable dysmorphisms and neurocognitive delay. With the exception of one maternally inherited deletion, all occurred de novo, and the smallest of these harboured a single gene, BMP2. In two individuals with additional features of Alagille syndrome, deletion of both JAG1 and BMP2 were identified. Deletion of this region has not been described as a copy number variant in the Database of Genomic Variants and has not been identified in 13 321 individuals from other cohort examined by array CGH in our laboratory. Our findings demonstrate a novel genomic disorder characterised by deletion of BMP2 with variable cognitive deficits and dysmorphic features and show that individuals bearing microdeletions in 20p12.3 often present with WPW syndrome.


The Human Cathelicidin Antimicrobial Peptide LL-37 and Mimics are Potential Anticancer Drugs.
Antimicrobial peptides (AMPs) play a critical role in innate host defense against microbial pathogens in many organisms. The human cathelicidin, LL-37, has a net positive charge and is amphiphilic, and can eliminate pathogenic microbes directly via electrostatic attraction toward negatively charged bacterial membranes. A number of studies have shown that LL-37 participates in various host immune systems, such as inflammatory responses and tissue repair, in addition to its antibacterial properties. Moreover, recent evidence suggests that it is also involved in the regulation of cancer. Indeed, previous studies have suggested that human LL-37 is involved in carcinogenesis via multiple reporters, such as FPR2 (FPRL1), epidermal growth factor receptor, and ERBb2, although LL-37 and its fragments and analogs also show anticancer effects in various cancer cell lines. This discrepancy can be attributed to peptide-based factors, host membrane-based factors, and signal regulation. Here, we describe the association between AMPs and cancer with a focus on anticancer peptide functions and selectivity in an effort to understand potential therapeutic implications.


The phenylpropanoid pathway is controlled at different branches by a set of R2R3-MYB C2 repressors in grapevine.
Because of the vast range of functions that phenylpropanoids possess, their synthesis requires precise spatiotemporal coordination throughout plant development and in response to the environment. The accumulation of these secondary metabolites is transcriptionally controlled by positive and negative regulators from the MYB and basic helix-loop-helix protein families. We characterized four grapevine (Vitis vinifera) R2R3-MYB proteins from the C2 repressor motif clade, all of which harbor the ethylene response factor-associated amphiphilic repression domain but differ in the presence of an additional TLLLFR repression motif found in the strong flavonoid repressor Arabidopsis (Arabidopsis thaliana) AtMYBL2. Constitutive expression of VvMYB4a and VvMYB4b in petunia (Petunia hybrida) repressed general phenylpropanoid biosynthetic genes and selectively reduced the amount of small-weight phenolic compounds. Conversely, transgenic petunia lines expressing VvMYBC2-L1 and VvMYBC2-L3 showed a severe reduction in petal anthocyanins and seed proanthocyanidins together with a higher pH of crude petal extracts. The distinct function of these regulators was further confirmed by transient expression in tobacco (Nicotiana benthamiana) leaves and grapevine plantlets. Finally, VvMYBC2-L3 was ectopically expressed in grapevine hairy roots, showing a reduction in proanthocyanidin content together with the down-regulation of structural and regulatory genes of the flavonoid pathway as revealed by a transcriptomic analysis. The physiological role of these repressors was inferred by combining the results of the functional analyses and their expression patterns in grapevine during development and in response to ultraviolet B radiation. Our results indicate that VvMYB4a and VvMYB4b may play a key role in negatively regulating the synthesis of small-weight phenolic compounds, whereas VvMYBC2-L1 and VvMYBC2-L3 may additionally fine tune flavonoid levels, balancing the inductive effects of transcriptional activators.


On the verge of extinction: genetics of the critically endangered Iberian plant species, Borderea chouardii (Dioscoreaceae) and implications for conservation management.
Borderea chouardii is a relictual and dioecious, strictly sexually reproducing, long-living geophyte of the Dioscoreaceae family. Previous biological and demographic studies have indicated the existence of a uniformly distributed panmictic population of this taxon at the southernmost Spanish pre-Pyrenean mountain ranges where it occurs in rather inaccessible crevices of a single limestone cliff. However, individuals of B. chouardii are spatially subdivided into two subpopulations located, respectively, on the upper and lower parts of the cliff, and vertically separated 150 m. Because of its extreme rarity, B. chouardii was the first Iberian taxon to have a specific conservation plan and has been included in several red lists under the category of critically endangered (CR). However, no previous attempts have been conducted to analyse the fine scale evolutionary mechanisms involved in its present microspatial distribution. Genetic diversity and population structure have been investigated through the analysis of neutral hypervariable markers such as simple sequence repeats (SSRs) and randomly amplified polymorphic DNAs (RAPDs) to unravel the impact of life history traits in the differentiation of the two subpopulations. Both types of molecular markers were unequivocal in distinguishing two genetically distinct groups of individuals corresponding to their spatial separation. However, SSRs detected a higher level of subpopulation differentiation (F(ST) = 0.35, R(ST) = 0.32) than RAPDs (F(ST) = 0.21). SSR data indicated significant deviation from random dispersal of genes and genotypes between the two groups, suggesting that mating occurs mainly among individuals within subpopulations, thus, favouring the divergence between the two groups. This microevolutionary differentiation scenario might have been caused by a coupled effect of past genetic drift and reproductive isolation, as a result of strong glacial age bottlenecks and inefficient dispersal system of pollen and seeds, respectively. The identification of such genetic structure in this narrow endemic prompts a modification of the management strategies of its single extant population.


Do estrogens enhance activation of brown and beiging of adipose tissues?
Obesity and its associated co-morbidities are worldwide public health concerns. Obesity is characterized by excessive adipose tissue accumulation; however, it is important to recognize that human and rodent adipose tissues are made up of several distinct adipose tissue sub-types. White adipose tissue (WAT) is considered the prototypical fat cell, due to its capacity and capability to store large amounts of lipid. In contrast, brown adipose tissue (BAT) oxidizes substrates to generate heat. BAT contains more mitochondria than WAT and express uncoupling protein-1 (UCP1), which mediates BAT thermogenesis. A third sub-type of adipose tissue, Brown-in-white (BRITE)/beige adipocytes arise from WAT upon adrenergic stimulation and resembles BAT functionally. The energy burning feature of BAT/beige cells, combined with evidence of an inverse-correlation between BAT/beige adipose tissue and obesity have given rise to the hypothesis that obesity may be linked to BAT/beige 'malfunction'. Females have more BAT and perhaps an enhanced capacity to beige their adipose tissue when compared to males. Multiple signal pathways are capable of activating BAT thermogenesis and beiging of WAT; here, we discuss the potential role of estrogens in enhancing and mediating these factors to enhance adipose tissue thermogenesis.


An eFTD-VP framework for efficiently generating patient-specific anatomically detailed facial soft tissue FE mesh for craniomaxillofacial surgery simulation.
Accurate surgical planning and prediction of craniomaxillofacial surgery outcome requires simulation of soft tissue changes following osteotomy. This can only be achieved by using an anatomically detailed facial soft tissue model. The current state-of-the-art of model generation is not appropriate to clinical applications due to the time-intensive nature of manual segmentation and volumetric mesh generation. The conventional patient-specific finite element (FE) mesh generation methods are to deform a template FE mesh to match the shape of a patient based on registration. However, these methods commonly produce element distortion. Additionally, the mesh density for patients depends on that of the template model. It could not be adjusted to conduct mesh density sensitivity analysis. In this study, we propose a new framework of patient-specific facial soft tissue FE mesh generation. The goal of the developed method is to efficiently generate a high-quality patient-specific hexahedral FE mesh with adjustable mesh density while preserving the accuracy in anatomical structure correspondence. Our FE mesh is generated by eFace template deformation followed by volumetric parametrization. First, the patient-specific anatomically detailed facial soft tissue model (including skin, mucosa, and muscles) is generated by deforming an eFace template model. The adaptation of the eFace template model is achieved by using a hybrid landmark-based morphing and dense surface fitting approach followed by a thin-plate spline interpolation. Then, high-quality hexahedral mesh is constructed by using volumetric parameterization. The user can control the resolution of hexahedron mesh to best reflect clinicians' need. Our approach was validated using 30 patient models and 4 visible human datasets. The generated patient-specific FE mesh showed high surface matching accuracy, element quality, and internal structure matching accuracy. They can be directly and effectively used for clinical simulation of facial soft tissue change.


Adult Mental Health, Substance Use Disorders, and Functional Outcomes of Children Resilient to Early Adversity.
Some children are unaffected by mental illness despite exposure to childhood adversity. These children are typically considered resilient. The objective of this study was to follow up such resilient children in adulthood to characterize mental health status, substance use, and functional outcomes. The analysis was based on the prospective, representative Great Smoky Mountains Study (N=1,420). Participants were assessed for psychiatric disorders and exposure to adversity with the structured Child and Adolescent Psychiatric Assessment interview up to eight times in childhood (ages 9-16; 6,674 observations). In total, 1,266 participants (86.3%) were followed up in adulthood at ages 25 and 30 to assess psychiatric disorders, substance use disorders, and functional outcomes. Seventy-five percent of the sample had met criteria for a psychiatric disorder or displayed subthreshold psychiatric problems by age 16. The number of adverse childhood experiences was strongly associated with childhood psychiatric status. Of children exposed to multiple adversities (N=650), 12.2% (N=63) did not display psychiatric problems. This group meets common definitions of childhood resilience. In adulthood, these individuals showing childhood resilience had greater risk of anxiety (risk ratio=2.9, 95% CI=1.0-9.1) and depressive (risk ratio=4.5, 95% CI=1.1-16.7) disorders, as well as worse physical health (means ratio=0.7, 95% CI=0.5-0.9) and financial or educational functioning (means ratio=0.6, 95% CI=0.5-0.7), compared with individuals exposed to fewer childhood adversities. These individuals showing childhood resilience did not have elevated risk for substance use disorders. Resilience to childhood adversity was uncommon. Individuals who appeared resilient in childhood were at risk for delayed poorer outcomes in adulthood. Public health efforts should prioritize minimizing early adversity exposure over promoting resilience.


Antiplasmodial activity of aqueous extract of Berberis aristata roots against Plasmodium berghei-infected BALB/c mice.
The rising problem of resistance to present antimalarial drugs stresses the need to look for newer antiplasmodial components with effective modes of action. The roots of Berberis aristata DC. (Berberidaceae) are used in the traditional medicine for malaria in various parts of India. The objective of this study was to evaluate antiplasmodial activity of B. aristata roots extract for the validation of its traditional medicinal use. Aqueous root extract of Berberis aristata (AREBA) was screened for its in vitro as well as in vivo antiplasmodial activity against lethal rodent malaria parasite Plasmodium berghei NK65. In vitro activity was evaluated against schizont maturation of P. berghei using various concentrations ranging from 1 to 100 µg/mL. For in vivo studies, AREBA at the doses of 150, 250, 350, and 650 mg/kg/d was administered to P. berghei infected BALB/c mice orally for 4 consecutive days (D0-D3). AREBA showed in vitro antiplasmodial activity with an IC50 value of 40 µg/mL. In vivo studies demonstrated a variable dose-dependent chemosuppression with higher efficacy at lower doses. At a dose of 350 mg/kg/d, the suppressive and preventive activities were found to be 67.1% and 53.9%, respectively, followed by enhancing mean survival period up to 12.8 d for the curative assay versus 7.5 d for the untreated mice. These results provide relevant scientific evidences for the traditional medicinal use of this plant as malaria remedy and further advocates the isolation and characterization of active antiplasmodial principle from this plant.


Decreasing Reimbursement of Pituitary Tumor Surgery: An Analysis of Medicare Data From 2010 to 2020.
Despite the extensive amount of research aimed at comparing patient outcomes between microscopic transsphenoidal surgery (MTSS) and endoscopic transsphenoidal surgery (ETSS) approaches, there has been relatively little recent investigation into the nationwide utilization and reimbursement trends of both techniques. This study aimed to identify trends in pituitary tumor surgery utilization, charges to Medicare, and reimbursement dependent on (1) MTSS/ETSS surgery type, (2) provider type (ie, neurosurgeon vs ear, nose, and throat), and (3) cosurgery status. This study used publicly available data from the Medicare Physician/Supplier Procedure Summary for the years 2010-2020. Linear regression was used to quantify temporal trends for submitted service counts, submitted charges, reimbursements, and reimbursement-to-charge across the 2010-2020 period. Regarding service count trends from 2010 to 2020, our results demonstrate a significant increase in ETSS utilization ( = 1.55, CI = 0.99-2.12, P < .001), a significant decrease in MTSS utilization ( = -0.86, CI = -1.21 to -0.51, P < .001), a significant increase in services submitted by otolaryngologists ( = 0.59, CI = 0.24-0.93, P = .0040), and a significant increase in cosurgeries ( = 1.03, CI = 0.24-0.93, P = .0051). Importantly, our results also demonstrated a significant decrease in reimbursements for ETSS procedures ( = -12.74, CI = -22.38 to -3.09, P = .015) and for pituitary tumor surgeries submitted by neurosurgeons specifically ( = -41.56, CI = -51.67 to -31.63, P < .0001). Our results demonstrated a significant increase in ETSS utilization and a significant decrease in MTSS utilization. We also noted a significant decrease in reimbursements for ETSS procedures and among procedures submitted by neurosurgeons specifically. We hope that our study highlights nationwide utilization and reimbursement patterns that may be useful for guiding future reimbursement-oriented policy development.


[Prednisolone sensitivity of peripheral blood mononuclear cells of patients with systemic lupus erythematosus clinically resistant to glucocorticoids and the effects of carbon hemoperfusion].
A study was made of the sensitivity to prednisolone (PS) of peripheral blood mononuclear cells (MS) of patients with systemic lupus erythematosus (SLE) with renal lesion showing resistance to glucocorticoids (GC). It was shown that in most of the patients (60%) MC were poorly inhibited by PS whereas the rest of the patients (40%) demonstrated in vitro sensitivity compared to that of the donors and even higher in some cases. Thus, the group of SLE patients showing clinical resistance to GC action was inhomogeneous in sensitivity of their MC to PS; only some patients showed resistance associated with the properties of the MC system. Hemocarboperfusion was shown to produce a considerable effect on MC sensitivity to PS: higher sensitivity was noted in all the cases, it was particularly noticeable in the group of patients with low basal sensitivity to PS. An analysis of the data obtained led to a conclusion that hemocarboperfusion raised MC sensitivity to PS. This mechanism was shown to determine the efficacy of the method of therapy of resistance to GC.


[Anatomic and functional ocular characteristics and myopia development].
Literature data dealing with the main anatomic and physiological factors influencing myopia development are reviewed. They are as follows: activity of accomodation structures, haemo- and hidrodynamic changes, rigidity of fibrous tunic. A complex of different examination methods is essential for appropriate management of myopic patients because the degree of influence is variable for every factor and is still completely unknown.


Challenges for environmental epidemiology research: are biomarker concentrations altered by kidney function or urine concentration adjustment?
Biomonitoring has become a standard approach for exposure assessment in occupational and environmental epidemiology. The use of biological effect markers to identify early adverse changes in target organs has also become widely adopted. However, the potential for kidney function to affect biomarker levels in the body and the optimal approach to adjustment of biomarker concentrations in spot urine samples for hydration status are two important but underappreciated challenges associated with biomarker use. Several unexpected findings, such as positive associations between urine nephrotoxicant levels and estimated glomerular filtration rate (eGFR), have been reported recently in research using biomarkers. These and other findings, discussed herein, suggest an impact of kidney glomerular filtration or tubule processing on biomarker levels. This is more commonly raised in the context of decreased kidney filtration, traditionally referred to as reverse causality; however, recent data suggest that populations with normal kidney filtration may be affected as well. Misclassification bias would result if biomarkers reflect kidney function as well as either exposures or early biological effect outcomes. Furthermore, urine biomarker associations with eGFR that differ markedly by approach used to adjust for urine concentration have been reported. Associations between urine measures commonly used for this adjustment, such as urine creatinine, and specific research outcomes could alter observed biomarker associations with outcomes. Research recommendations to address the potential impact of kidney function and hydration status adjustment on biomarkers are provided, including a range of approaches to study design, exposure and outcome assessment, and adjustment for urine concentration.


Development of a rapid HPLC-UV method for simultaneous quantification of protodioscin and rutin in white and green asparagus spears.
Asparagus (Asparagus officinalis L.) spears are rich in bioactive compounds such as protodioscin, a saponin, and rutin, a flavonoid. Protodioscin and rutin are routinely quantified separately, and an approach permitting simultaneous measurement would significantly improve speed of analysis. We have optimized an extraction procedure and modified a method of high-performance liquid chromatography by coupling to an ultraviolet detector to simultaneously analyze protodioscin and rutin in asparagus extracts. An acidic ethanol solvent was more efficient than methanol, acetonitrile, or water in coextraction of protodioscin and rutin. Protodioscin and rutin were detected at 210 nm, with retention times of 12.6 min and 7.9 min, respectively. The method was validated by high linear correlations between 3.13 and 1000.0 μg/mL for protodioscin (r(2)= 0.9999), and between 0.3 and 1087.5 μg/mL for rutin (r(2)= 0.9997). The limit(s) of detection and quantification for protodioscin were 1.6 μg/mL and 3.13 μg/mL, respectively, and for rutin 0.2 μg/mL and 0.3 μg/mL, respectively. White asparagus spears and the crown of the plants were revealed to be rich sources of protodioscin and contained 2.59 to 10.4 mg/g dry weight. Green asparagus spears, particularly the upper portion, were rich in rutin and contained between 1.51 and 7.29 mg/g dry weight.


Playing it safe: The relationship between parent attitudes to risk and injury, and children's adventurous play and physical activity.
Children naturally seek risk in play and adventurous play outdoors confers many benefits, including the potential to increase moderate-to-vigorous physical activity (MVPA). This study aimed to investigate the relationship between parent attitudes to risk and injury, and their elementary school-aged child's daily adventurous play and MVPA. A panel sample of 645 Australian parents/guardians completed an online survey consisting of several validated measures of risk and injury attitudes, and physical activity and play behaviour. Data were analysed via descriptive statistics, univariate and multivariable regressions using Stata 17. A series of exploratory univariate logistic regressions were conducted, followed by a series of multivariable logistic regressions fitted to test the association between parent risk and injury attitudes and (i) children's MVPA, (ii) active play and (iii) adventurous play, while adjusting for socio-demographic factors. Most adult participants (81%) were female. The mean age of the child participants (53% male) was 8.6 years (SD = 2.4). On average, parents were positive about children's engagement with risk, however, 78% of parents had low tolerance of risk when presented with specific play scenarios, and attitudes towards injuries varied, with mothers more concerned than fathers. After adjusting for confounders, children with parents who were tolerant of risk in play were more likely to meet the MVPA guideline of ≥60 min daily (OR 2.86, CI: 1.41, 5.82, p < 0.004) and spend more time playing adventurously (OR 3.03, CI: 1.82, 5.06, p < 0.001). Positive associations for MVPA and adventurous play were observed across all models examining parent attitudes to risk and injury. Younger children engaged in more play and physical activity, however, more positive parent attitudes appeared to moderate the age-related influences. We found a divergence between the outcomes parents desire for their children through engagement with risk and the play activities they are comfortable with in practice. Parent attitudes to risk and injury are potentially modifiable factors that may increase children's affordances for adventurous play and physical activity. Interventions that provide parents with practical approaches to address injury concerns and support children's risk-taking in play outdoors are recommended.


Relationship between SARS-CoV-2 infection and the incidence of ventilator-associated lower respiratory tract infections: a European multicenter cohort study.
Although patients with SARS-CoV-2 infection have several risk factors for ventilator-associated lower respiratory tract infections (VA-LRTI), the reported incidence of hospital-acquired infections is low. We aimed to determine the relationship between SARS-CoV-2 pneumonia, as compared to influenza pneumonia or no viral infection, and the incidence of VA-LRTI. Multicenter retrospective European cohort performed in 36 ICUs. All adult patients receiving invasive mechanical ventilation > 48 h were eligible if they had: SARS-CoV-2 pneumonia, influenza pneumonia, or no viral infection at ICU admission. VA-LRTI, including ventilator-associated tracheobronchitis (VAT) and ventilator-associated pneumonia (VAP), were diagnosed using clinical, radiological and quantitative microbiological criteria. All VA-LRTI were prospectively identified, and chest-X rays were analyzed by at least two physicians. Cumulative incidence of first episodes of VA-LRTI was estimated using the Kalbfleisch and Prentice method, and compared using Fine-and Gray models. 1576 patients were included (568 in SARS-CoV-2, 482 in influenza, and 526 in no viral infection groups). VA-LRTI incidence was significantly higher in SARS-CoV-2 patients (287, 50.5%), as compared to influenza patients (146, 30.3%, adjusted sub hazard ratio (sHR) 1.60 (95% confidence interval (CI) 1.26 to 2.04)) or patients with no viral infection (133, 25.3%, adjusted sHR 1.7 (95% CI 1.2 to 2.39)). Gram-negative bacilli were responsible for a large proportion (82% to 89.7%) of VA-LRTI, mainly Pseudomonas aeruginosa, Enterobacter spp., and Klebsiella spp. The incidence of VA-LRTI is significantly higher in patients with SARS-CoV-2 infection, as compared to patients with influenza pneumonia, or no viral infection after statistical adjustment, but residual confounding may still play a role in the effect estimates.


Analysis of the difference in the perinatal mortality rate between white-collar and blue-collar workers in Japan, 1995-2015.
This study investigated differences in the perinatal mortality rate between white-collar and blue-collar workers. Data from the "Report of Vital Statistics: Occupational and Industrial Aspects" in Japan covering the period from 1995 to 2015 were used. Five-year maternal age groups from 15-19 years to 45-49 years were analyzed according to work type, and the perinatal mortality rate for each age group and the age-standardized perinatal mortality rate according to maternal age were calculated in each analyzed year. A Bayesian age-period-cohort analysis was used to estimate age, period, and cohort effects for the perinatal mortality rate according to work type. Moreover, the perinatal mortality rate ratios between types of workers were estimated for each age group, period, and cohort. The estimated perinatal mortality rate ratios of blue-collar to white-collar workers were above 1 in most of the age groups and cohorts. The age effect for the perinatal mortality rate among white-collar workers was the largest in the 15-year to 19-year age group, whereas that among blue-collar workers was the largest in the 45-year to 49-year age group. Furthermore, the estimated perinatal rate ratio between white-collar and blue-collar workers tended to increase with maternal age. The magnitude of the decrease of the cohort effects on the perinatal mortality rate was rather larger in blue-collar workers in the cohorts born between 1946-1950 and 1996-2000. The magnitude of the disparity markedly increased with maternal age. Thus, middle-aged blue-collar workers need more prenatal care and preventive measures for perinatal mortality than white-collar workers.


Control of gene expression during higher plant chloroplast biogenesis. Protein synthesis and transcript levels of psbA, psaA-psaB, and rbcL in dark-grown and illuminated barley seedlings.
Etioplasts of 4.5-day-old dark-grown barley synthesize and accumulate most of the membrane and nearly all the soluble polypeptides of mature chloroplasts of light-grown seedlings. Etioplasts do not synthesize a limited set of chloroplast-encoded polypeptides which are major constituents of chloroplast thylakoid membranes: two chlorophyll apoproteins of photosystem I (68 and 65 kDa), two chlorophyll apoproteins of photosystem II (47 and 43 kDa), and a 32-kDa polypeptide which has now been identified as the psbA gene product. Throughout development in the dark, etioplasts were unable to synthesize the chlorophyll apoproteins of photosystem I and II or the psbA gene product despite the presence of significant transcript levels for psbA and psaA-psaB (encode for photosystem I chlorophyll apoproteins). Light was not required for the synthesis of ribulose bisphosphate carboxylase large subunit with the highest rate of large subunit synthesis occurring in young dark-grown seedlings. Illumination of 4.5-day-old dark-grown barley rapidly induced the synthesis of the chlorophyll apoproteins and the psbA gene product at a time when transcript levels for psbA and psaA-psaB did not increase appreciably. Therefore, during the early stages of light-induced development the synthesis of the chlorophyll apoproteins of photosystem I and psbA gene product is regulated at the translational level. With continued chloroplast development in the light, the synthesis of the chlorophyll apoproteins of photosystem I and II decline rapidly as does the synthesis of the large subunit of ribulose bisphosphate carboxylase. The decline in polypeptide synthesis correlated with a decline in rbcL and psaA-psaB transcript levels and a light-dependent decline in plastid rRNA content. In contrast, synthesis of the psbA gene product was maintained throughout light-induced chloroplast development which correlated with the maintenance of psbA transcript levels. However, light is not strictly required for psbA transcript accumulation since psbA transcript levels increased slightly with continued plastid development in dark-grown seedlings.


p53-induced inhibition of protein synthesis is independent of apoptosis.
Activation of a temperature-sensitive form of p53 in murine erythroleukaemia cells results in a rapid impairment of protein synthesis that precedes inhibition of cell proliferation and loss of cell viability by several hours. The inhibition of translation is associated with specific cleavages of polypeptide chain initiation factors eIF4GI and eIF4B, a phenomenon previously observed in cells induced to undergo apoptosis in response to other stimuli. Although caspase activity is enhanced in the cells in which p53 is activated, both the effects on translation and the cleavages of the initiation factors are completely resistant to inhibition of caspase activity. Moreover, exposure of the cells to a combination of the caspase inhibitor z-VAD.FMK and the survival factor erythropoietin prevents p53-induced cell death but does not reverse the inhibition of protein synthesis. We conclude that the p53-regulated cleavages of eIF4GI and eIF4B, as well as the overall inhibition of protein synthesis, are caspase-independent events that can be dissociated from the induction of apoptosis per se.


How Language Is Embodied in Bilinguals and Children with Specific Language Impairment.
This manuscript explores the role of embodied views of language comprehension and production in bilingualism and specific language impairment. Reconceptualizing popular models of bilingual language processing, the embodied theory is first extended to this area. Issues such as semantic grounding in a second language and potential differences between early and late acquisition of a second language are discussed. Predictions are made about how this theory informs novel ways of thinking about teaching a second language. Secondly, the comorbidity of speech, language, and motor impairments and how embodiment theory informs the discussion of the etiology of these impairments is examined. A hypothesis is presented suggesting that what is often referred to as specific language impairment may not be so specific due to widespread subclinical motor deficits in this population. Predictions are made about how weaknesses and instabilities in speech motor control, even at a subclinical level, may disrupt the neural network that connects acoustic input, articulatory motor plans, and semantics. Finally, I make predictions about how this information informs clinical practice for professionals such as speech language pathologists and occupational and physical therapists. These new hypotheses are placed within the larger framework of the body of work pertaining to semantic grounding, action-based language acquisition, and action-perception links that underlie language learning and conceptual grounding.


[Role of Mechanical Bowel Preparation for Elective Colorectal Surgery].
The presence of bowel contents during colorectal surgery has been related to surgical site infections (SSI), anastomotic leakage (AL) and postoperative complications theologically. Mechanical bowel preparation (MBP) for elective colorectal surgery aims to reduce fecal materials and bacterial count with the objective to decrease SSI rate, including AL. Based on many observational data, meta-analysis and multicenter randomized control trials (RTC), non-MBP did not increase AL rates or SSI and other complications in colon and even rectal surgery. In 2011 Cochrane review, there is no significant benefit MBP compared with non-MBP in colon surgery and also no better benefit MBP compared with rectal enemas in rectal surgery. However, in surgeon's perspectives, MBP is still in widespread surgical practice, despite the discomfort caused in patients, and general targeting of the colon microflora with antibiotics continues to gain popularity despite the lack of understanding of the role of the microbiome in anastomotic healing. Recently, there are many evidence suggesting that MBP+oral antibiotics (OA) should be the growing gold standard for colorectal surgery. However, there are rare RCT studies and still no solid evidences in OA preparation, so further studies need results in both MBP and OA and only OA for colorectal surgery. Also, MBP studies in patients with having minimally invasive surgery (MIS; laparoscopic or robotics) colorectal surgery are still warranted. Further RCT on patients having elective left side colon and rectal surgery with primary anastomosis in whom sphincter saving surgery without MBP in these MIS and microbiome era.


Determinants of self-reported mental health using the British household panel survey.
The study of self-reported mental health is a fairly recent area for economists, although sociologists, psychologists and public health specialists have been studying it for years. One methodological problem with earlier research is that there are many unobserved characteristics of individuals that may be correlated with self-reported mental health. Neglecting these factors may lead to biased estimates of the effects of variables such as income, education, health, etc. Panel data enables us to control for unobserved individual specific effects, whereas a cross-section study or time series study cannot. This paper examines the determinants of self-reported mental health in UK using data from the first eight waves of the British Household Panel Survey. In particular, we are interested in assessing the effect of education on self-reported mental health which other studies have ignored. The measure of self-reported mental health used in this paper is the General Health Questionnaire (GHQ). To account for the possible correlation between the unobserved individual effects and some explanatory variables, a Hausman Taylor's instrumental variables estimator (HT) is employed. In order to derive this estimator, one has to distinguish between variables that are correlated with the individual specific effects (endogenous) and variables which are uncorrelated with the individual specific effects (exogenous). This HT estimator also allows for estimating the parameters corresponding with time invariant variables such as education and ethnicity. The evidence presented here confirms that mental health scores mentioned on the GHQ are significantly related to job status, age, marital status and self-assessed health status. The results also show no evidence that income impacts on self-reported mental health. Ethnicity is also found to deteriorate self-reported mental health yet the effect is not significant. The results of this paper also show that education had no significant impact on self-reported mental health. IMPLICATIONS FOR MENTAL HEALTH POLICY: Issues related to unemployment and social cohesion may be relevant factors in the prevention of mental illness. Policies aimed at improving these factors have an impact on the mental health status of society. In consideration of the evidence of gender differential in mental health, mental health policies should take into account properly this issue. In order to draw definite conclusions, it is important to formally test the presence of attrition bias as well as expand the sample to include more waves. Still, we are concerned about the issue of weak correlation between the instruments and potential endogenous variables. Additionally, we have to bear in mind that inconsistent estimates may potentially occur if the partition of the variables in subsets of endogenous and exogenous is not correctly specified. These issues need further research. The estimation technique also presented in this paper may be applied to a wide range of health services research.


More Than 30 000-fold Field Enhancement of Terahertz Nanoresonators Enabled by Rapid Inverse Design.
The rapid development of 6G communications using terahertz (THz) electromagnetic waves has created a demand for highly sensitive THz nanoresonators capable of detecting these waves. Among the potential candidates, THz nanogap loop arrays show promising characteristics but require significant computational resources for accurate simulation. This requirement arises because their unit cells are 10 times smaller than millimeter wavelengths, with nanogap regions that are 1 000 000 times smaller. To address this challenge, we propose a rapid inverse design method using physics-informed machine learning, employing double deep Q-learning with an analytical model of the THz nanogap loop array. In ∼39 h on a middle-level personal computer, our approach identifies the optimal structure through 200 000 iterations, achieving an experimental electric field enhancement of 32 000 at 0.2 THz, 300% stronger than prior results. Our analytical model-based approach significantly reduces the amount of computational resources required, offering a practical alternative to numerical simulation-based inverse design for THz nanodevices.


Unraveling the Activity Trends and Design Principles of Single-Atom Catalysts for Nitrate Electrocatalytic Reduction.
Electrocatalytic nitrate (NO3-) reduction represents one of the most promising approaches to mitigate NO3- pollution and yield NH3, but it is still challenged by the atomic economy and selectivity issues of substantial active sites. Here, we describe a comprehensive investigation on a series of single-atom catalysts (SACs) using nitrogen-doped carbon as substrate (metal/NC). The essence of activity is related to the extent of the electron transfer capacity (SAs → NO3-). Among these examined SACs, the Cu/NC presents good performance toward NH3 synthesis, i.e., a maximum NH3 Faradaic efficiency of 100% with a high NH3 yield rate of up to 32,300 μg h-1 mgcat.-1. X-ray absorption fine structure spectra and density functional theory calculations provide evidence that the electronic structure of Cu-N4 coordination prohibits the formation of N2, N2O, and H2 and facilitates the orbital hybridization between the 2p orbitals of NO3- and 3d orbitals of Cu single-atom sites. Our study is believed to provide fundamental guidance for the future design of highly efficient electrocatalysts in NO3- reduction to NH3.


Isolation of Bacillus licheniformis var. endoparasiticus from the blood of rheumatoid arthritis patients and normal subjects.
Sixty-five samples of blood from patients with rheumatoid arthritis and 94 from control subjects were examined by multiple culture and monthly subculture for periods of up to 33 months to detect the revision stages of Bacillus licheniformis var. endoparasiticus (BLE), which exists as L-forms in blood. Isolation of BLE were obtained more often from the blood of control subjects than from rheumatoid patients during the first 6 months of incubation, when there was clustering of positive cultures within samples. Thereafter, the isolation rate was similar for the two groups and positive cultures were distributed randomly between samples. Isolation of diphtheroid intermediate reversion stages (phase A and B) occurred mainly during the first year of incubation, but isolations of the fully reverted sporing bacillus (phase C) increased in frequency with incubation time, particularly from cultures with a high rate of desiccation during prolonged incubation. The proportion of different phases of BLE amongst the isolates and the distribution of the phases with incubation time were similar for the rheumatoid-arthritis patients and the normal subjects.


Inter-organ substrate exchanges in the critically ill.
Metabolic inter-organ exchange is a major field of research for improving the treatment of the critically ill. Adapting regional blood flows is the first regulatory step, although the relationships between hypoperfusion and metabolic disorders are matter of controversy. Metabolic steady state results from a vast inter-organ interplay and several nutrients or metabolites are signalling molecules in the regulation of gene transcription. Inter- or intra-organ substrate recycling shares or delays the mandatory need for aerobic ATP synthesis in some conditions. Nitrogen metabolism is highly compartmentalised in an inter-organ co-operation and liver, muscle, kidney and gut are the most important organs. By remodelling the amino acid mixture delivered to peripheral cells after intestinal absorption, the liver plays a determinant role in whole body protein synthesis. Albumin turnover increases after brain injury. Since the location of synthesis is different to that of breakdown this turnover can be viewed as an inter-organ exchange. The metabolic side of pH homeostasis is also an inter-organ exchange mainly shared by liver, kidney and muscle.


Risk factors and outcomes for the acquisition of carbapenem-resistant Gram-negative bacillus bacteremia: A retrospective propensity-matched case control study.
A substantial number of carbapenem-resistant Gram-negative bacilli (CR GNB) have been identified among the etiologic multidrug-resistant GNB in healthcare-associated infections. For achieving a better therapeutic outcome by minimizing inappropriate empirical antibiotic treatment before blood culture and susceptibility testing results are available, it is very important to identify patients who are at risk for the development of CR GNB bacteremia. Retrospective analysis of propensity-score matched (PSM) adult patients with CR GNB bacteremia (PSM-group 1 [n = 95]) and those with non-CR GNB bacteremia (PSM-group 2 [n = 190]). PSM-group 1 was found to a significantly longer length of hospital stay (27 vs. 18 days; p < 0.001) after emerging GNB bacteremia and a higher 30-day all-cause mortality rate (27.4% vs. 5.8%; p < 0.001), when compared with PSM-2 group. Independent risk factors for the acquisition of CR GNB bacteremia were previous exposure to an antipseudomonal penicillin (odds ratio [OR] = 3.58; 95% confidence interval [CI] = 1.30-9.90), an antipseudomonal cephalosporin (OR = 3.49; 95% CI = 1.09-11.24), and a carbapenem (OR = 3.60; 95% CI = 1.37-9.47), and longer length of hospital stay before the development of GNB bacteremia (OR = 1.03; 95% CI = 1.01-1.05). Risk factors for acquisition of CR GNB bacteremia identified in this study each may serve as a reminder alerting clinicians to hospitalized patients at risk for CR GNB bacteremia requiring appropriate antibiotic coverage, and in these circumstances, combined antibiotics may be used until antimicrobial de-escalation/adjustment is clearly indicated by the subsequently identified pathogenic GNB and its susceptibility profile.


Mesenchymal Stromal Cells Improve Renovascular Function in Polycystic Kidney Disease.
Polycystic kidney disease (PKD) is a common cause of end-stage renal failure, for which there is no accepted treatment. Progenitor and stem cells have been shown to restore renal function in a model of renovascular disease, a disease that shares many features with PKD. The objective of this study was to examine the potential of adult stem cells to restore renal structure and function in PKD. Bone marrow-derived mesenchymal stromal cells (MSCs, 2.5 × 10(5)) were intrarenally infused in 6-week-old PCK rats. At 10 weeks of age, PCK rats had an increase in systolic blood pressure (SBP) versus controls (126.22 ± 2.74 vs. 116.45 ± 3.53 mmHg, p < 0.05) and decreased creatinine clearance (3.76 ± 0.31 vs. 6.10 ± 0.48 µl/min/g, p < 0.01), which were improved in PKD animals that received MSCs (SBP: 114.67 ± 1.34 mmHg, and creatinine clearance: 4.82 ± 0.24 µl/min/g, p = 0.001 and p = 0.003 vs. PKD, respectively). MSCs preserved vascular density and glomeruli diameter, measured using microcomputed tomography. PCK animals had increased urine osmolality (843.9 ± 54.95 vs. 605.6 ± 45.34 mOsm, p < 0.01 vs. control), which was improved after MSC infusion and not different from control (723.75 ± 56.6 mOsm, p = 0.13 vs. control). Furthermore, MSCs reduced fibrosis and preserved the expression of proangiogenic molecules, while cyst size and number were unaltered by MSCs. Delivery of exogenous MSCs improved vascular density and renal function in PCK animals, and the benefit was observed up to 4 weeks after a single infusion. Cell-based therapy constitutes a novel approach in PKD.


Free Energy Analyses of Cell-Penetrating Peptides Using the Weighted Ensemble Method.
Cell-penetrating peptides (CPPs) have been widely used for drug-delivery agents; however, it has not been fully understood how they translocate across cell membranes. The Weighted Ensemble (WE) method, one of the most powerful and flexible path sampling techniques, can be helpful to reveal translocation paths and free energy barriers along those paths. Within the WE approach we show how Arg9 (nona-arginine) and Tat interact with a DOPC/DOPG(4:1) model membrane, and we present free energy (or potential mean of forces, PMFs) profiles of penetration, although a translocation across the membrane has not been observed in the current simulations. Two different compositions of lipid molecules were also tried and compared. Our approach can be applied to any CPPs interacting with various model membranes, and it will provide useful information regarding the transport mechanisms of CPPs.


Serum proinsulin in children and adolescents with chemical diabetes.
Proinsulin levels (PLC) in serum were determined after gel filtration on specimens obtained during oral glucose tolerance testing (OGTT) in seven patients with repeated abnormality in OGTT and in a group of seven control subjects matched for age. Fasting and thirty-, sixty-, and 120-minute postglucose venous samples were analyzed for glucose (PG) and immunocreactive insulin (IRI) as well as PLC, PG and IRI mean concentrations were greater at all testing times in the patient population, but the mean IRI/PG was significantly higher in the patients only at fasting. PLC mean levels were higher in the patients but not to a level of significance. Percentage of total IRI atributable to PLC at each time point was identical between the two groups. The apparcent diminished effectiveness of circulating insulin in chemical diabetes cannot be attributed to an abnormal proportion of proinsulin.


Flow cytometry sorting of nuclei enables the first global characterization of Paramecium germline DNA and transposable elements.
DNA elimination is developmentally programmed in a wide variety of eukaryotes, including unicellular ciliates, and leads to the generation of distinct germline and somatic genomes. The ciliate Paramecium tetraurelia harbors two types of nuclei with different functions and genome structures. The transcriptionally inactive micronucleus contains the complete germline genome, while the somatic macronucleus contains a reduced genome streamlined for gene expression. During development of the somatic macronucleus, the germline genome undergoes massive and reproducible DNA elimination events. Availability of both the somatic and germline genomes is essential to examine the genome changes that occur during programmed DNA elimination and ultimately decipher the mechanisms underlying the specific removal of germline-limited sequences. We developed a novel experimental approach that uses flow cell imaging and flow cytometry to sort subpopulations of nuclei to high purity. We sorted vegetative micronuclei and macronuclei during development of P. tetraurelia. We validated the method by flow cell imaging and by high throughput DNA sequencing. Our work establishes the proof of principle that developing somatic macronuclei can be sorted from a complex biological sample to high purity based on their size, shape and DNA content. This method enabled us to sequence, for the first time, the germline DNA from pure micronuclei and to identify novel transposable elements. Sequencing the germline DNA confirms that the Pgm domesticated transposase is required for the excision of all ~45,000 Internal Eliminated Sequences. Comparison of the germline DNA and unrearranged DNA obtained from PGM-silenced cells reveals that the latter does not provide a faithful representation of the germline genome. We developed a flow cytometry-based method to purify P. tetraurelia nuclei to high purity and provided quality control with flow cell imaging and high throughput DNA sequencing. We identified 61 germline transposable elements including the first Paramecium retrotransposons. This approach paves the way to sequence the germline genomes of P. aurelia sibling species for future comparative genomic studies.


Evolution of population dynamics following invasion by a non-native predator.
Invasive predatory species are frequently observed to cause evolutionary responses in prey phenotypes, which in turn may lead to evolutionary shifts in the population dynamics of prey. Research has provided a link between rates of predation and the evolution of prey population growth in the lab, but studies from natural populations are rare. Here, we tested for evolutionary changes in population dynamics parameters of zooplankton Daphnia pulicaria following invasion by the predator Bythotrephes longimanus into Lake Kegonsa, Wisconsin, US. We used a resurrection ecological approach, whereby clones from pre- and post-invasive periods were hatched from eggs obtained in sediment cores and were used in a 3-month growth experiment. Based on these data, we estimated intrinsic population growth rates (r), the shape of density dependence (θ) and carrying capacities (K) using theta-logistic models. We found that post-invasion Daphnia maintained a higher r and K under these controlled, predation-free laboratory conditions. Evidence for changes in θ was weaker. Whereas previous experimental evolution studies of predator-prey interactions have demonstrated that genotypes that have evolved under predation have inferior competitive ability when the predator is absent, this was not the case for the Daphnia. Given that our study was conducted in a laboratory environment and the possibility for genotype-by-environment interactions, extrapolating these apparent counterintuitive results to the wild should be done with caution. However, barring such complications, we discuss how selection for reduced predator exposure, either temporally or spatially, may have led to the observed changes. This scenario suggests that complexities in ecological interactions represents a challenge when predicting the evolutionary responses of population dynamics to changes in predation pressure in natural systems.


Noninvasive radioisotopic assessment of pulmonary vascular protein leak. Experimental studies and potential clinical applications.
Noninvasive, double-radioisotope measurement of pulmonary vascular protein leak is a specific and sensitive method for assessing pulmonary vascular permeability. It is sensitive enough to detect increases in vascular protein leak that are too small to produce detectable increases in gravimetric lung water. A simple method for calculating protein leak index reliably reflects expected changes in pulmonary vascular permeability in a broad range of lung injury conditions. This method of measurement may have significant experimental and clinical applications in the study of lung-injury-induced edema, and especially in the evaluation of levels of injury associated with increased permeability but not significant edema. In particular, this method may help to distinguish between increased-permeability edema, such as that which occurs in the adult respiratory distress syndrome, and other types of pulmonary edema in humans.


Characterization of a novel CYP2C9 mutation (1009C>A) detected in a warfarin-sensitive patient.
Warfarin is the most frequently prescribed anticoagulant for the long-term treatment in the clinic. Recent studies have shown that polymorphic alleles within the CYP2C9, VKORC1, and CYP4F2 genes are related to the warfarin dosage requirement. In this study, a novel non-synonymous mutation (1009C>A) in CYP2C9 was detected in a warfarin-hypersensitive patient, while the other two candidate genes were both found to be homozygous for the wild-type alleles. The newly identified point mutation results in an amino acid substitution at position 337 of the CYP2C9 protein (P337T) and has been designated as the novel allele CYP2C9*58. When expressed in insect cell microsomes, the relative intrinsic clearance values of the CYP2C9.58 variant for tolbutamide and losartan were quite similar to those of the typical defective variant CYP2C9.3, whereas the clearance value of CYP2C9.58 for diclofenac was slightly higher than that of another typical defective variant CYP2C9.2. These data suggested that when compared with wild-type CYP2C9.1, the enzymatic activity of the novel allelic variant has been greatly reduced by the 1009C>A mutation. If patients carrying this allele take drugs metabolized by CYP2C9, their metabolic rate might be slower than that of wild-type allele carriers and thus much more attention should be paid to their clinical care.


Sodium-potassium--dependent ATPase. II. Cytochemical localization during the reversal of galactose cataracts in rat.
The importance of enzyme Na-K-ATPase in the development of galactose-induced cataractogenesis is now well realized. In our recent studies we reported decreased level of activity of this enzyme with an increased duration of galactose feeding and the induced alterations in rat ocular lens. Our approach was to determine the level of Na-K-ATPase activity by ultrastructural cytochemical analysis of lenses and spectrophotometric analysis of the incubating media used for cytochemical localization as described by Ernst. Using these approaches, we have determined the activity level of this enzyme during the reversal phase of the galactose-induced injury to the lens. Our findings are presented in this report and show that the activity of Na-K-ATPase recovers rapidly and attains the normal level when the animals were transferred to Rat Chow diet after the establishment of mature cataracts resulting from galactose feeding. This study supports the previous biochemical and morphological findings that partial reversal of galactose-induced cataractous lens occurs upon discontinuation of feeding of cataractogenic agent galactose.