Comparative growth performance of backgrounded beef heifers treated with an injectable fixed-dose combination (0.2 mg/kg doramectin + 6.0 mg/kg levamisole hydrochloride) or single-active (0.2 mg/kg ivermectin) endectocide.
Gastrointestinal nematodes (GINs) can negatively impact all production classes of cattle, particularly growing cattle. A global decline in efficacy of broad-spectrum single-active anthelmintics requires alternative GIN control methods without the aid of novel drug classes. Here, we present a new fixed-dose combination injectable (FDCI) endectocide for cattle that combines doramectin (5 mg/ml) and levamisole hydrochloride (150 mg/ml). A 56-day comparative performance confinement backgrounding trial was conducted in stocker beef heifers (n = 1548) with confirmed GIN infections to (1) compare the Day 14 post-treatment effectiveness of the new FDCI endectocide to pen mates treated with the injectable single-active endectocide ivermectin, as evidenced by fecal egg counts (FECs) conducted for a randomly selected subset (10%) of both treatment groups, and (2) determine if the greater GIN control by the FDCI evidenced in the subsample improved growth performance in all FDCI-treated heifers. Heifers were procured in four cohorts, with a 10-week timeframe between enrollment of the first and last cohort. Treatment groups were comingled within dirt-floor pens (n = 31; 7-8 per cohort) and offered a standard backgrounding diet ad libitum for the study duration. Heifers with enrollment FEC ≥ 30 eggs per gram (EPG) were randomly allocated to receive the FDCI (n = 773) or ivermectin (n = 775) on Day 0. Day 0 FECs conducted on 10% of enrolled heifers (FDCI, n = 78; ivermectin, n = 79) were not different between treatment groups (p = 0.491). Day 14 FECs for the same heifers were reduced compared to Day 0 within each treatment group. Heifers given the FDCI had lower Day 14 AM FECs and higher FEC reduction test (FECRT) result (0.07 EPG; 0.999) than ivermectin-treated heifers (21.58 EPG; FECRT = 0.850). Mean body weight (BW) was not different between treatment groups on Day 0 (p = 0.2762) and Day 14 (p = 0.2010) but was significantly greater (p = 0.0007) for FDCI-treated heifers compared to ivermectin-treated heifers on Day 56. Compared to ivermectin-treated heifers, overall average daily gain from all evaluation periods (Day 0-14, Day 14-56, and Day 0-56) was greater (p ≤ 0.0052) in FDCI-treated heifers, and FDCI-treated heifers had 4.223 kg greater total weight gain over the 56-day study. The FDCI (0.2 mg/kg doramectin + 6.0 mg/kg levamisole hydrochloride) was highly effective in reducing GIN infections and thus promoted improved growth performance in beef heifers over a 56-day backgrounding period.


High prevalence of gastroesophageal reflux in vocal opera students. A case-control type study.
Opera singers are at risk to develop gastroesophageal reflux, because of the vocational challenges during singing. The singers are reluctant to be submitted to pH-metry fearing throat injuries. We evaluated the gastroesophageal reflux in vocal opera students using a non-invasive salivary test. A prospective, case-control study was carried out in 30 vocal opera students from the Music Academy of the city Cluj-Napoca, Romania, enrolled in the "canto" section. For control, 20 medical students and residents were enrolled. Each subject filled in a questionnaire and gave saliva samples for the salivary test Peptest™ (RD Biomed). The statistical analysis was made using program R version 3.2.1 (2015-06-18). According to salivary Peptest, vocal opera students had a higher prevalence of gastro-esophageal reflux than the control group (96.67% versus 30%; 29 from 30 vs 14 from 20), P<0.001. Only 50% (15 of 30) of the vocal opera students reported reflux symptoms. Salivary Peptest confirmed a higher prevalence of gastroesophageal reflux in vocal opera students, although not all of them reported reflux symptoms.


Radiomics Analysis Based on Diffusion Kurtosis Imaging and T2 Weighted Imaging for Differentiation of Pancreatic Neuroendocrine Tumors From Solid Pseudopapillary Tumors.
To develop and validate a radiomics model of diffusion kurtosis imaging (DKI) and T2 weighted imaging for discriminating pancreatic neuroendocrine tumors (PNETs) from solid pseudopapillary tumors (SPTs). Sixty-six patients with histopathological confirmed PNETs (n = 31) and SPTs (n = 35) were enrolled in this study. ROIs of tumors were manually drawn on each slice at T2WI and DWI (b = 1,500 s/mm2) from 3T MRI. Intraclass correlation coefficients were used to evaluate the interobserver agreement. Mean diffusivity (MD) and mean kurtosis (MK) were derived from DKI. The least absolute shrinkage and selection operator regression were used for feature selection. MD and MK had a moderate diagnostic performancewith the area under curve (AUC) of 0.71 and 0.65, respectively. A radiomics model, which incorporated sex and age of patients and radiomics signature of the tumor, showed excellent discrimination performance with AUC of 0.97 and 0.86 in the primary and validation cohort. Moreover, the new model had better diagnostic performance than that of MD (P = 0.023) and MK (P = 0.004), and showed excellent differentiation with a sensitivity of 95.00% and specificity of 91.67% in primary cohort, and the sensitivity of 90.91% and specificity of 81.82% in the validation cohort. The accuracy of radiomics analysis, radiologist 1, and radiologist 2 for diagnosing SPTs and PNETs were 92.42, 77.27, and 78.79%, respectively. The accuracy of radiomics analysis was significantly higher than that of subjective diagnosis (P < 0.05). Radiomics model could improve the diagnostic accuracy of SPTs and PNETs and contribute to determining an appropriate treatment strategy for pancreatic tumors.


The mechanism of immunoinhibition by arachidonic and linoleic acid: effects on the lymphoid and reticulo-endothelial systems.
Subcutaneous injection with certain polyunsaturated fatty acids (PUFA), in particular linoleic acid (C18:2), has recently been shown to prolong the survival of skin allografts in mice and to reduce both primary and secondary cytotoxic responses by isolated spleen cells. In this study we have examined changes in the lymphoid and reticulo-endothelial systems of both grafted and ungrafted mice treated according to the schedules shown to prolong allograft survival. Many of the changes in the lymphoid and reticulo-endothelial systems which can be produced by allografting could also be produced in ungrafted animals by PUFA treatment. These changes included: increased 125IudR uptake by spleen, lymph nodes, and bone marrow, increased spleen and lymph node weight, increased proportion of red pulp and granulocyte precursors in the spleen, and reticulo-endothelial activation as shown by an increased rate of carbon clearance. The combined effects of allografting and C18:2 on peripheral lymph node weight were greater than the effect of either treatment alone, but fell short of the sum of both treatments. Allografting increased 125IudR uptake/unit organ weight, not only in the peripheral lymph nodes, but also in bone marrow, thymus and spleen. When allografted animals were C18:2 treated, this increase was considerably reduced, uptake being little different from that in C18:2 treated ungrafted controls. The number of theta-positive cells in the spleen was markedly increased by a tumor allograft, but such an increase could not be seen in allografted animals when treated with C18:2. Whilst prolonged C18:2 treatment led to destructive changes in the spleen, immunoinhibition could still be demonstrated after shorter treatments with C18:2 when no gross histological evidence for tissue destruction was apparent. Partial reversibility of inhibition of cytotoxic activity of isolated spleen cells could be domonstrated when treatment was discontinued.


Biomechanical Analysis of the Closed Kinetic Chain Upper-Extremity Stability Test.
The closed kinetic chain upper-extremity stability (CKCUES) test is a functional test for the upper extremity performed in the push-up position, where individuals support their body weight on 1 hand placed on the ground and swing the opposite hand until touching the hand on the ground, then switch hands and repeat the process as fast as possible for 15 s. To study scapular kinematic and kinetic measures during the CKCUES test for 3 different distances between hands. Experimental. Laboratory. 30 healthy individuals (15 male, 15 female). Participants performed 3 repetitions of the test at 3 distance conditions: original (36 in), interacromial, and 150% interacromial distance between hands. Participants completed a questionnaire on pain intensity and perceived exertion before and after the procedures. Scapular internal/external rotation, upward/downward rotation, and posterior/anterior tilting kinematics and kinetic data on maximum force and time to maximum force were measured bilaterally in all participants. Percentage of body weight on upper extremities was calculated. Data analyses were based on the total numbers of hand touches performed for each distance condition, and scapular kinematics and kinetic values were averaged over the 3 trials. Scapular kinematics, maximum force, and time to maximum force were compared for the 3 distance conditions within each gender. Significance level was set at α = .05. Scapular internal rotation, posterior tilting, and upward rotation were significantly greater in the dominant side for both genders. Scapular upward rotation was significantly greater in original distance than interacromial distance in swing phase. Time to maximum force in women was significantly greater in the dominant side. CKCUES test kinematic and kinetic measures were not different among 3 conditions based on distance between hands. However, the test might not be suitable for initial or mild-level rehabilitation due to its challenging requirements.


Arginine Homopeptide of 11 Residues as a Model of Cell-Penetrating Peptides in the Interaction with Bacterial Membranes.
Cell-penetrating peptides rich in arginine are good candidates to be considered as antibacterial compounds, since peptides have a lower chance of generating resistance than commonly used antibiotics. Model homopeptides are a useful tool in the study of activity and its correlation with a secondary structure, constituting an initial step in the construction of functional heteropeptides. In this report, the 11-residue arginine homopeptide (R11) was used to determine its antimicrobial activity against Staphylococcus aureus and Escherichia coli and the effect on the secondary structure, caused by the substitution of the arginine residue by the amino acids Ala, Pro, Leu and Trp, using the scanning technique. As a result, most of the substitutions improved the antibacterial activity, and nine peptides were significantly more active than R11 against the two tested bacteria. The cell-penetrating characteristic of the peptides was verified by SYTOX green assay, with no disruption to the bacterial membranes. Regarding the secondary structure in four different media-PBS, TFE, E. coli membrane extracts and DMPG vesicles-the polyproline II structure, the one of the parent R11, was not altered by unique substitutions, although the secondary structure of the peptides was best defined in E. coli membrane extract. This work aimed to shed light on the behavior of the interaction model of penetrating peptides and bacterial membranes to enhance the development of functional heteropeptides.


Epithelial-mesenchymal transition via transforming growth factor beta in pancreatic cancer is potentiated by the inflammatory glycoprotein leucine-rich alpha-2 glycoprotein.
We previously showed that an inflammation-related, molecule leucine-rich alpha-2 glycoprotein (LRG) enhances the transforming growth factor (TGF)-β1-induced phosphorylation of Smad proteins and is elevated in patients with pancreatic ductal adenocarcinoma (PDAC). As TGF-β/Smad signaling is considered to play a key role in epithelial-mesenchymal transition (EMT), we attempted to clarify the mechanism underlying LRG-related EMT in relation to metastasis in PDAC. We cultured LRG-overexpressing PDAC cells (Panc1/LRG) and evaluated the morphology, EMT-related molecules and TGF-β/Smad signaling pathway in these cells. We also assessed the LRG levels in plasma and resected specimens from patients with PDAC. Inflammatory cytokines induced LRG production in PDAC cells. A spindle-like shape was visualized more frequently than other shapes in Panc1/LRG with TGF-β1 exposure. The expression of E-cadherin in Panc1/LRG was decreased with TGF-β1 exposure. Invasion increased with TGF-β1 stimulation of Panc1/LRG. The phosphorylation of smad2 in Panc1/LRG was increased in comparison with parental Panc1 under TGF-β1 stimulation. In the plasma LRG-high group, the recurrence rate tended to be higher and the recurrence-free survival (RFS) tended to be worse in comparison with the plasma LRG-low group. LRG enhanced EMT induced by TGF-β signaling, thus indicating that LRG has a significant effect on the metastasis of PDAC.


Presence of an acute phase response in sheep with clinical classical scrapie.
Work with experimental scrapie in sheep has been performed on-site for many years including studies on PrPSc dissemination and histopathology of organs and tissues both at preclinical and clinical stages. In this work serum was sampled at regular intervals from lambs which were infected immediately after birth and from parallel healthy controls, and examined for acute phase proteins. In contrast to earlier experiments, which extensively studied PrPSc dissemination and histopathology in peripheral tissues and brain, this experiment is focusing on examination of serum for non-PrPSc markers that discriminates the two groups, and give insight into other on-going processes detectable in serum samples. There was clear evidence of an acute phase response in sheep with clinical scrapie, both experimental and natural. All the three proteins, ceruloplasmin, haptoglobin and serum amyloid A, were increased at the clinical stage of scrapie. There was evidence of a systemic measurable acute phase response at the clinical terminal end-stage of classical scrapie.


Controlled delivery of vascular endothelial growth factor promotes neovascularization and maintains limb function in a rabbit model of ischemia.
Vascular endothelial growth factor (VEGF) modulates new blood vessel development and growth and has been suggested as a potential therapeutic agent that could alleviate debilitating claudication in patients. The objective of this study was to determine whether controlled, local delivery of a low dose of VEGF from an osmotic pump could promote neovascularization, limb perfusion, and functional improvements in the hind limbs of rabbits rendered partially ischemic by surgery. The effects of VEGF were compared with those of the vasodilator nitroglycerin (NTG) and to saline administered similarly. Thirty rabbits were randomly assigned to either VEGF (n = 10), NTG (n = 10), or saline (n = 10) treatment groups. Partial ischemia was induced in each left hind limb by surgical ligation of the common and superficial femoral arteries, leaving the internal iliac artery intact. The right limb of each animal served as a nonischemic control. Immediately after vessel ligations, a 28-day osmotic pump was implanted to deliver VEGF (0.22 microg/kg/day), NTG (17.8 microg/kg/day), or saline solution into the common iliac artery just proximal to the ligation site. Comparative vascularity between ischemic and nonischemic limbs within treatment groups and between groups was evaluated by (1) capillary counts from representative fields of hematoxylin and eosin stained muscle tissue taken from hind limbs at day 40; (2) digitized arteriograms of ischemic legs at day 40, which were used to quantify the complexity of vascular branching (fractal dimension index) and the total extent of vascularization (vascular density index); (3) measuring capillary refill times in ischemic limbs; and (4) observations of functional and trophic changes in ischemic limbs. Statistical differences between treatment groups were evaluated by one-way ANOVA. Complexity of vascular branching and vascular density were significantly greater (p < 0.001) in VEGF-treated ischemic limbs compared with NTG- and saline-treated ischemic limbs. By postoperation day 14, all VEGF-treated ischemic limbs had restored capillary refill (p < 0.001), new hair growth, and greatly improved limb function and appearance. Saline-treated limbs exhibited ischemic changes, with poor capillary refill and negligible limb function. Capillary refill in NTG-treated ischemic limbs did not differ significantly from saline-treated limbs. Ischemic VEGF-treated limbs had significantly more capillaries compared with both ischemic and nonischemic limbs in saline-treated animals (p < 0.05). Ischemic NTG-treated limbs also had significantly more capillaries compared with ischemic limbs in saline-treated animals (p < 0.05). Because of high variability, however, capillary counts in VEGF-treated ischemic limbs did not differ significantly from those of contralateral nonischemic limbs, or from capillary counts in either ischemic or nonischemic limbs of NTG-treated rabbits. Controlled release of microgram quantities of VEGF significantly enhanced neovascularization and vascular perfusion in ischemic limbs compared with controls in this rabbit model of partial ischemia. In addition, VEGF-treated ischemic limbs demonstrated near-normal function and appearance, whereas NTG- and saline-treated ischemic controls remained noticeably impaired. This novel approach of VEGF delivery may prove clinically useful either alone or combined with revascularization procedures.


Parasite infections in nestling red-shouldered hawks (Buteo lineatus) in northeast Wisconsin.
Red-shouldered hawks (Buteo lineatus) are threatened in Wisconsin and long-term data suggest that nest productivity is low in the state for unknown reasons. Our objective was to determine whether red-shouldered hawks in northeast Wisconsin were infected with parasites that could contribute to low nest productivity. We examined nestlings for the presence of Trichomonas gallinae, Protocalliphora avium, and blood parasites in June 2006 and 2007. We did not detect T. gallinae in throat swabs taken from 24 nestlings in 2007. Ear canals of nestlings were parasitized by P. avium larvae in 10 of 11 (91%) nests and in 22 of 24 (92%) nestlings. Larvae were found in higher intensity in 1 ear relative to the other. Leucocytozoon toddi was present in 90.5% (38/42) of the nestlings. At least 1 bird in each nest was infected. Intensity of L. toddi averaged 48.6 +/- 58.3 infected cells per 2,000 erythrocytes (2.4 +/- 2.9%). No other blood parasites were identified.


Mexican-Origin Youth's Cultural Orientations and Values: Do Older Sisters and Brothers Matter?
Family is an important context for cultural development, but little is known about the contributions of siblings. This study investigated whether older siblings' cultural orientations and familism values predicted changes in younger siblings' cultural orientations and familism values across 2 years and tested sibling characteristics and younger siblings' modeling as moderators. Participants were 246 Mexican-origin younger (Mage = 17.72; SD = 0.57) and older siblings (Mage = 20.65; SD = 1.57) and their parents. Findings revealed that older siblings' Anglo orientations and familism values interacted with younger siblings' modeling: When younger siblings reported high modeling, older siblings' Anglo orientations and values predicted increases in younger siblings' Anglo orientations and values. Discussion highlights the importance of siblings in cultural socialization.


Diagnosis of Osteoarthritis by Cartilage Surface Smoothness Quantified Automatically from Knee MRI.
We investigated whether surface smoothness of articular cartilage in the medial tibiofemoral compartment quantified from magnetic resonance imaging (MRI) could be appropriate as a diagnostic marker of osteoarthritis (OA). At baseline, 159 community-based subjects aged 21 to 81 with normal or OA-affected knees were recruited to provide a broad range of OA states. Smoothness was quantified using an automatic framework from low-field MRI in the tibial, femoral, and femoral subcompartments. Diagnostic ability of smoothness was evaluated by comparison with conventional OA markers, specifically cartilage volume from MRI, joint space width (JSW) from radiographs, and pain scores. A total of 140 subjects concluded the 21-month study. Cartilage smoothness provided diagnostic ability in all compartments (P < 0.0001). The diagnostic smoothness markers performed at least similar to JSW and were superior to volume markers (e.g., the AUC for femoral smoothness of 0.80 was higher than the 0.57 for volume, P < 0.0001, and marginally higher than 0.73 for JSW, P = 0.25). The smoothness markers allowed diagnostic detection of pain presence (P < 0.05) and showed some correlation with pain severity (e.g., r = -0.32). The longitudinal change in smoothness was correlated with cartilage loss (r up to 0.60, P < 0.0001 in all compartments). This study demonstrated the potential of cartilage smoothness markers for diagnosis of moderate radiographic OA. Furthermore, correlations between smoothness and pain values and smoothness loss and cartilage loss supported a link to progression of OA. Thereby, smoothness markers may allow detection and monitoring of OA-supplemented currently accepted markers.


Paleo-oscillomics: inferring aspects of Neanderthal language abilities from gene regulation of neural oscillations.
Language seemingly evolved from changes in brain anatomy and wiring. We argue that language evolution can be better understood if particular changes in phasal and cross-frequency coupling properties of neural oscillations, resulting in core features of language, are considered. Because we cannot track the oscillatory activity of the brain from extinct hominins, we used our current understanding of the language oscillogenome (that is, the set of genes responsible for basic aspects of the oscillatory activity relevant for language) to infer some properties of the Neanderthal oscillome. We have found that several candidates for the language oscillogenome show differences in their methylation patterns between Neanderthals and humans. We argue that differences in their expression levels could be informative of differences in cognitive functions important for language.


Alpha (1,2)-fucosyltransferase M307A polymorphism improves piglet survival.
To confirm the beneficial effects of alpha (1,2)-fucosyltransferase (FUT1) M307 (A) on piglet survival on commercial farms, we performed PCR-RFLP analysis of FUT1 M307 in successfully marketed (n = 245) and disease affected/deceased pigs during weaning (n = 252) at a commercial farm. We also evaluated the FUT1 genotypes of 190 healthy pigs from three different genetic backgrounds. The distribution of genotypes differed between the successfully marketed and disease affected/deceased pig groups. The frequency of the A allele, associated with resistance to edema and post-weaning diarrhea, was higher in the post-weaning survival group (0.21) than in the non-survival group (0.16, P < 0.05). The odds ratio for piglet survival between AA and GG genotypes was 1.98; thus, piglet survival for individuals with the AA genotype was almost two-fold greater than for GG individuals. The FUT1 gene polymorphism can be used as an effective marker for selection programs to improve post-weaning piglet survival.


[Immunologic aspects related to hemoglobinopathic diseases].
The records of 96 patients with sickle cell disease were studied in relation to various immunological parameters. Age, sex, levels of total and fractionated serum protein and immunoglobulins, serum complements, antierythrocytic antibodies, circulating immune complexes, antiviral antibodies and lymphoid populations were taken into account. No major alterations in humoral immunity were found in this group of panamanian sickle cell patient.


Psoriatic fasciitis in a pediatric patient: A case report.
Diffuse fasciitis with psoriatic arthritis on magnetic resonance imaging (MRI) has not been previously described in childhood. Here we present the first case report of a pediatric patient developing fasciitis, beyond plantar fasciitis, with psoriatic arthritis. An 11-year-old female was admitted with the complaints of psoriatic rash on the body associated with severe pain in the lower extremities and arthritis in the right knee. Psoriasis was confirmed by skin biopsy, she diagnosed with juvenile psoriatic arthritis. Diagnostic tests did not indicate any pathology except MRI. MRI of the femur and tibia revealed that high-signal inflammatory changes in the subdermal fascia. These findings led to a diagnosis of psoriatic fasciitis. Methotrexate was given for 3 mo but the patient showed no response to therapy; therefore, etanercept was added. However, there was no response to treatment. Etanercept was switched to adalimumab at the sixth month of therapy. Clinical improvement started with therapy of adalimumab within one month. Fasciitis finding in MRI disappeared at seventh months on adalimumab therapy. She has no complaint for two years with adalimumab. The most effective imaging method is MRI and adalimumab may be the best choice of treatment for psoriatic fasciitis.


Relationship between clinical and radiologic findings of spinal cord injury in decompression sickness.
Decompression sickness may involve the central nervous system. The most common site is spinal cord. This study was conducted to determine the relationship between magnetic resonance(MR) imaging findings of spinal damage. We conducted a retrospective review of 12 patients (male=10, female=2) who presented with spinal cord symptoms. We investigated their clinical features, neurological findings and radiologic findings. The depth and bottom time of the dive were 34.5 meters (range 22-56) and 22.7 minutes (range 10-55) respectively. Most divers ascended within appropriate time frame as shown by the decompression tables. The most frequent initial symptoms were lower limb weakness (n=12), followed by sensory disturbances (n=10) and bladder dysfuction (n=5). The chief radiologic abnormalities were continuous (n=3), or non-continuous (n=5) high-signal intensity on T2-weighted images at posterior paramedian portion of the spinal cord, mainly thoracic level. There were no abnormal findings in the remaining four (4) patients, and they showed good prognosis. All patients were treated with hyperbaric oxygen therapy and some received high-dose dexamethasone. On discharge, five (5) patients had made a full recovery, seven (7) had some residual neurological sequelae, and all patients except one (1) regained normal bladder function. Spinal cord decompression sickness is a neurological emergency. Early recognition and treatment may minimize neurological damage. Initial normal finding in MR imaging was a good predictor for prognosis in spinal decompression sickness.


Medical treatment of patients with infantile spasms.
Infantile spasms are the main feature in West syndrome, an age-related epilepsy syndrome that affects 1 in every 2,000-4,000 infants. The authors provide a comprehensive review of the literature about infantile spasms and their therapy. In the United States, the drug of choice for infantile spasms, at least the cryptogenic cases, has been adrenocorticotropic hormone (ACTH). It is generally considered to be more effective than corticosteroids. Adrenocorticotropic hormone appears to alter long-term prognosis of cryptogenic infantile spasms, and helps in some cases of symptomatic infantile spasm. Vigabatrin has been considered the drug of choice for infantile spasms secondary to tuberous sclerosis, and possibly, according to many neurologists, for all cases of infantile spasm. Recent concerns regarding retinopathy associated with vigabatrin therapy are, however, limiting the use of this drug. Valproic acid benefits 40%-70% of patients who failed a trial of ACTH. Nitrazepam is as effective as ACTH in acutely controlling infantile spasms; however, its long-term effects on prognosis have not been studied. Pyridoxine, lamotrigine, topiramate, zonisamide, ketogenic diet, immunoglobulin therapy, felbamate, and thyrotropin-releasing hormone have all been used for the treatment of infantile spasms, but are usually reserved for cases refractory to vigabatrin and/or ACTH.


[Cesarean section in a patient with threatening hemoptysis caused by tracheal carcinoid].
A woman 29 weeks pregnant presented with acute respiratory insufficiency and massive hemoptysis of unknown origin. An emergency cesarean section was performed to avoid hypoxic fetal damage during episodes of maternal hypoxemia. Bleeding was due to an unsuspected endotracheal carcinoid tumor located near the carina. The tumor obstructed the distal portion of the trachea, leading to life-threatening complications during tracheal intubation because of its histological characteristics and placement. Pulse oximetry response to standard mechanical ventilation was unsatisfactory. Special measures to ventilate mother and fetus were successful, in spite of iatrogenic pneumothorax and pneumomediastinum. We report an exceptional case of tracheal carcinoid tumor during the pregnancy and recommend that tracheal or bronchial intubation take place under fiber optic guidance in cases of hemoptysis with no firm etiologic diagnosis.


Expert Consensus on SABA Use for Asthma Clinical Decision-Making: A Delphi Approach.
A modified Delphi process was undertaken to provide a US expert-led consensus to guide clinical action on short-acting beta2-agonist (SABA) use. This comprised an online survey (Phase 1), forum discussion and statement development (Phase 2), and statement adjudication (Phase 3). In Phase 1 (n = 100 clinicians), 12% routinely provided patients with ≥4 SABA prescriptions/year, 73% solicited SABA use frequency at every patient visit, and 21% did not consult asthma guidelines/expert reports. Phase 3 experts (n = 8) reached consensus (median Likert score, interquartile range) that use of ≥3 SABA canisters/year is associated with increased risk of exacerbation and asthma-related death (5, 4.75-5); SABA use history should be solicited at every patient visit (5, 4.75-5); usage patterns over time, not absolute thresholds, should guide response to SABA overuse (5, 4.5-5). Future asthma guidelines should include clear recommendations regarding SABA usage, using expert-led thresholds for action.


A proangiogenic signature is revealed in FGF-mediated bevacizumab-resistant head and neck squamous cell carcinoma.
Resistance to antiangiogenic therapies is a critical problem that has limited the utility of antiangiogenic agents in clinical settings. However, the molecular mechanisms underlying this resistance have yet to be fully elucidated. In this study, we established a novel xenograft model of acquired resistance to bevacizumab. To identify molecular changes initiated by the tumor cells, we performed human-specific microarray analysis on bevacizumab-sensitive and -resistant tumors. Efficiency analysis identified 150 genes upregulated and 31 genes downregulated in the resistant tumors. Among angiogenesis-related genes, we found upregulation of fibroblast growth factor-2 (FGF2) and fibroblast growth factor receptor-3 (FGFR3) in the resistant tumors. Inhibition of the FGFR in the resistant tumors led to the restoration of sensitivity to bevacizumab. Furthermore, increased FGF2 production in the resistant cells was found to be mediated by overexpression of upstream genes phospholipase C (PLCg2), frizzled receptor-4 (FZD4), chemokine [C-X3-C motif] (CX3CL1), and chemokine [C-C motif] ligand 5 (CCL5) via extracellular signal-regulated kinase (ERK). In summary, our work has identified an upregulation of a proangiogenic signature in bevacizumab-refractory HNSCC tumors that converges on ERK signaling to upregulate FGF2, which then mediates evasion of anti-VEGF therapy. These findings provide a new strategy on how to enhance the therapeutic efficacy of antiangiogenic therapy. Novel xenograft model leads to the discovery of FGF as a promising therapeutic target in overcoming the resistance of antiangiogenic therapy in HNSCC.


Localized myopathy in a young man with abetalipoproteinaemia-myocardial infarction?
Myocardial infarction has not previously been reported as a complication of abetalipoproteinaemia (ABL) We describe a case of probable asymptomatic myocardial infarction in a 24-year-old male with ABL and no previous cardiac history . Electrocardiographic and imaging evidence of the injury is presented.


Antioxidant capacity and phenolic composition of leaves from 10 Bene (Pistacia atlantica subsp. kurdica) genotypes.
Leaves of 10 Bene genotypes were collected from six provinces (West Azerbaijan, Kurdistan, Kermanshah, Ilam, Hamedan and Lorestan) in Iran. This study was carried out to better characterise the total phenolic and flavonoid contents, antioxidant capacity and phenolic composition, as well as to evaluate the correlation between content of phenolic compounds and antioxidant activity in 10 genotypes of Bene. The antioxidant activity of leaf extracts was measured using different assays: ferric reducing antioxidant power, nitric oxide radical scavenging and 2,2-diphenyl-1-picrylhydrazyl. The results indicated that the antioxidant capacity can be related to total phenolic and flavonoid content, so that among all the genotypes studied here, the highest and the lowest phenolic content and antioxidant activity were observed in B2 and B10 genotypes, respectively. Analysing the phenolic composition using high performance liquid chromatography, caffeic acid, vanillic acid, ferulic acid and sinapic acid were found in all investigated genotypes.


Resolving morphology and antibody labeling over large distances in tissue sections.
Protein expression patterns are a primary determinant of tissue function and in this study we developed methods to study protein expression over macroscopic distances at subcellular levels of detail. Using the mammalian lens as a model tissue system, we show that by combining two-photon microscopy with novel image montage methods (fast beam blanking coupled with mathematical alignment tools) we have extended the limited field of view of laser scanning microscopes. To illustrate the utility of our approach, the distribution of connexin-46 was visualized across equatorial sections of the rat mammalian lens. By optimizing fixation protocols, good morphological preservation could be achieved over the thickness of the lens (approximately 4 mm) while preserving antigenicity of lens proteins. Using the same image data, changes in lens fiber cell morphology were mapped quantitatively by automatic image analysis routines. The methods presented should be generally applicable to any tissue system where changes in antibody labeling and tissue structure occur over large and small distances.


Assessment of treatment-related thyroid dysfunction in patients with head and neck cancer.
To assess thyroid dysfunction in head and neck cancer patients who have received external beam radiotherapy according to radiotherapy fields and dose, tumor site and other local or systemic treatments retrospectively and prospectively and propose a follow-up schedule. A total of 378 patients was classified into two groups. Group I (n = 345) consisted of surgically treated 153 laryngeal, 80 nasopharyngeal and 112 oral cavity/oropharyngeal carcinoma patients; these patients were evaluated retrospectively for treatment-related thyroid dysfunction using their data files. Group II included 33 patients with head and neck cancer who were evaluated prospectively. Thyroid function tests were performed at the beginning of the radiotherapy and every three months after the radiotherapy course, and thyroid dysfunction regarding surgery, radiotherapy and chemotherapy was evaluated. In Group I, the median follow-up for 153 operated laryngeal carcinoma patients was 44 months. Four (2.6%) of them were found to have clinically apparent hypothyroidism. After a median follow-up of 36 months, none of the 80 nasopharyngeal carcinoma patients showed signs of hypothyroidism. Clinically apparent hypothyroidism was detected in only 1 (0.8%) of the oral cavity/oropharyngeal carcinoma patients after a median follow-up of 25 months. In Group II, 1 (3%) patient was found to have thyroid dysfunction postoperatively prior to radiotherapy. At the time of analysis, 29 (87.8%) patients were euthyroidic, 2 (6.1%) patients had subclinical and 2 (6.1%) patients had clinical hypothyroidism. All patients with thyroid dysfunction have had combined surgery and radiotherapy, and none of the patients treated with radical radiotherapy has experienced hypothyroidism. Even after a short follow-up, the incidence of thyroid dysfunction was 12.2% in head and neck cancer patients treated with combined surgery and radiotherapy. We recommend thyroid function tests in these patients prior to and once every 3-6 months after the radiotherapy course.


Long-term follow-up of patients with chronic myeloid leukemia in chronic phase developing sudden blast phase on imatinib therapy.
Sudden blast phase (SBP) is a rare event that occurs in an unpredictable fashion amongst patients with chronic myeloid leukemia (CML) who otherwise appear to be responding satisfactorily to imatinib (IM) treatment. We investigated the incidence, clinical characteristics, treatment outcome and long-term follow-up of 213 patients with chronic phase CML treated with IM according to the European LeukemiaNet guidelines. Nine patients, eight of whom received IM as first-line therapy, developed SBP (4.2% of the total). They tended to have low or intermediate risk Sokal scores at diagnosis, a predominance of the lymphoid phenotype and a short interval from "optimal" response to the development of BP. Five of the nine patients with SBP are alive in complete molecular remission; however, all of them underwent allogeneic hematopoietic stem cell transplant. The cumulative incidence of SBP for the patients who received IM as first-line therapy was 5.9% and the 2-year overall survival of the nine patients who developed SBP was 56%. Despite the improved outcome for patients with SBP receiving tyrosine kinase inhibitors (TKIs) and transplant, many of these patients are not salvaged with these therapies. This illustrates the need to develop predictive models to identify patients early whose response to TKI therapy will not be durable and hopefully prevent the transformation to advanced disease.


Hyperkinetic Movement Disorders in Children: A Brief Review.
Movement disorders are common neurologic disturbances in childhood. There are two major types of movement disorders. Hypokinetic disorders are with paucity of voluntary movements and are very uncommon in pediatric age group. Hyperkinetic movement abnormalities are very common in children and defined as abnormal repetitive involuntary movements. Movement disorders in childhood and even in adolescents are different in etiology, timing, treatment and prognosis versus adulthood movement abnormalities. In this brief article, we reviewed common types of hyperkinetic abnormal movements in children and adolescents with emphasis on etiologies, new classifications and recent treatment strategies.


Myosin-like sequences in the malaria parasite Plasmodium falciparum bind human erythrocyte membrane protein 4.1.
Plasmodium falciparum malaria is one of the most lethal infectious diseases afflicting humanity. During development within the erythrocyte, P. falciparum induces significant modifications to the structure and function of the human erythrocyte membrane. This study focused on the identification of new protein-protein interactions between host and parasite. A novel application of in vitro display technology was used: P. falciparum phage display expression libraries were screened against purified human erythrocyte protein 4.1. DNA sequencing and bioinformatic analyses were used to identify parasite proteins that bind protein 4.1. P. falciparum proteins displaying strong binding specificity toward protein 4.1 included five hypothetical proteins, erythrocyte binding antigen-175, erythrocyte binding ligand-1 like protein and a putative serine/threonine kinase. A common binding motif displaying homology to muscle myosin and neurofilament sequences was also identified in four of the eight proteins. These proteins are potentially involved in the invasion and/or release, as well as the growth and survival of malaria parasites during development with the red blood cell. The characterization of novel protein interactions between P. falciparum and erythrocyte membrane protein 4.1 will lead to a better understanding of malaria pathogenesis and parasite biology.


Storage of methane and freon by interstitial van der Waals confinement.
A known host-guest assembly, organized only by means of relatively weak dispersive forces, exhibits hitherto unappreciated thermal stability. The hexagonal close-packed arrangement of calix[4]arene contains lattice voids that can occlude small, highly volatile molecules. This host-guest system can be exploited to retain a range of freons, as well as methane, not only well above their normal boiling points, but also at relatively high temperatures and low pressures. The usually overlooked van der Waals interactions in organic crystals can indeed be used in a highly stable supramolecular system for gas storage.


Detection and HER2 expression of circulating tumor cells: prospective monitoring in breast cancer patients treated in the neoadjuvant GeparQuattro trial.
This study was aimed at detecting and characterizing circulating tumor cells (CTC) before and after neoadjuvant therapy (NT) in the peripheral blood of patients with breast cancer. The clinical trial GeparQuattro incorporated NT approaches (epirubicin/cyclophosphamide prior to randomization to docetaxel alone, docetaxel in combination with capecitabine, or docetaxel followed by capecitabine) and additional trastuzumab treatment for patients with HER2-positive tumors. We used the Food and Drug Administration-approved CellSearch system for CTC detection and evaluation of HER2 expression and developed HER2 immunoscoring for CTC. We detected > or =1 CTC/7.5 mL in 46 of 213 patients (21.6%) before NT and in 22 of 207 patients (10.6%) after NT (P = 0.002). Twenty (15.0%) initially CTC-positive cases were CTC-negative after NT, whereas 11 (8.3%) cases were CTC-positive after NT, although no CTC could be found before NT. CTC detection did not correlate with primary tumor characteristics. Furthermore, there was no association between tumor response to NT and CTC detection. HER2-overexpressing CTC were observed in 14 of 58 CTC-positive patients (24.1%), including 8 patients with HER2-negative primary tumors and 3 patients after trastuzumab treatment. CTC scored HER2-negative or weakly HER2-positive before or after NT were present in 11 of 21 patients with HER2-positive primary tumors. HER2 overexpression on CTC was restricted to ductal carcinomas and associated with high tumor stage (P = 0.002). CTC number was low in patients with primary breast cancer. The decrease in CTC incidence during treatment was not correlated with standard clinical characteristics and primary tumor response. Information on the HER2 status of CTC might be helpful for stratification and monitoring of HER2-directed therapies.


Gene-activated matrices for cartilage defect reparation.
A cartilage defect has a very limited ability to repair itself spontaneously due to the shortage of blood. Many attempts have been made to restore the integrity of cartilage in clinical and experimental studies. Recently, tissue engineering has emerged as a new protocol for lost tissue regeneration. Meanwhile, the defect-repairing environment can be improved by gene therapy methods. Gene-activated matrices (GAM) fabricated with biomaterials and plasmids fill the cartilage defects to restore the integrity of joint surface, facilitating repair cell adhesion and proliferation as well as the synthesis of extracelluar matrix. And they also serve as a local gene delivery system, inducing therapeutic agent expression at the repair site. In the present study, we fabricated two- and three-dimensional matrices from chitosan and gelatin, then added a plasmid DNA encoding transforming growth factors-ss1 (TGF-ss1) for cartilage defect regeneration. First, we demonstrated primary chondrocytes could maintain their biological characteristics and secrete therapeutic proteins when they were cultured onto GAM in vitro. Subsequently we inserted three-dimensional GAM into cartilage defects of rabbit knee joints. With the results of the new cartilage tissue formation, we came to the conclusion that GAM was helpful for new tissue production and this therapeutic protocol provided a cheap, simple, and effective method for cartilage defect reparation.


Urinary magnesium loss in aging diabetic mellitus rats.
Past studies on urinary loss of magnesium (Mg) have focused on young diabetic rats. The aims of the present study were to determine the rapidity at which glycosuria and magnesuria occur after the induction of diabetes mellitus (DM) in old male rats, and the maximal amount of Mg and glucose (Glu) loss in the urine and whether or not the loss is persistent and (3) the most sensitive means of correlating the Mg and Glu loss in the urine. Three methods of expressing urinary Mg and Glu concentrations were selected: Method A: mg/24 h; Method B: mg/24 h/kg body weight (BW), and Method C: ratio of Mg to creatinine (Mg/Crea) or ratio of Glu to Crea (Glu/Crea). Our study indicated a maximal and rapid loss of Mg and Glu occurring within 1 week after induction of DM (by streptozotocin injection) and remained in effect for 6 weeks, the end of the study. The urinary Mg loss due to DM correlated best with urinary Glu when expressed as mg/24 h/kg BW. In addition, the increase in urinary Mg concentration paralleled the degree of glycosuria and reached a maximum of 5-12 times baseline values when expressed by Method B. This was 4-10 times when expressed by Method A, and 2-6 times when expressed by Method C. Since polyuria is a feature of DM, we also correlated the relationship between these two factors, and found a significantly positive correlation (r = 0.735) particularly when Mg was expressed as mg/24 h. In summary, rapid and significant urinary Mg loss is observed in old rats made diabetic with streptozotocin.(ABSTRACT TRUNCATED AT 250 WORDS)


Methods and tools for spatial mapping of single-cell RNAseq clusters in Drosophila.
Single-cell RNA sequencing (scRNAseq) experiments provide a powerful means to identify clusters of cells that share common gene expression signatures. A major challenge in scRNAseq studies is to map the clusters to specific anatomical regions along the body and within tissues. Existing data, such as information obtained from large-scale in situ RNA hybridization studies, cell type specific transcriptomics, gene expression reporters, antibody stainings, and fluorescent tagged proteins, can help to map clusters to anatomy. However, in many cases, additional validation is needed to precisely map the spatial location of cells in clusters. Several approaches are available for spatial resolution in Drosophila, including mining of existing datasets, and use of existing or new tools for direct or indirect detection of RNA, or direct detection of proteins. Here, we review available resources and emerging technologies that will facilitate spatial mapping of scRNAseq clusters at high resolution in Drosophila. Importantly, we discuss the need, available approaches, and reagents for multiplexing gene expression detection in situ, as in most cases scRNAseq clusters are defined by the unique coexpression of sets of genes.


Synthetic hydrogel nanoparticles for sepsis therapy.
Sepsis is a life-threatening condition caused by the extreme release of inflammatory mediators into the blood in response to infection (e.g., bacterial infection, COVID-19), resulting in the dysfunction of multiple organs. Currently, there is no direct treatment for sepsis. Here we report an abiotic hydrogel nanoparticle (HNP) as a potential therapeutic agent for late-stage sepsis. The HNP captures and neutralizes all variants of histones, a major inflammatory mediator released during sepsis. The highly optimized HNP has high capacity and long-term circulation capability for the selective sequestration and neutralization of histones. Intravenous injection of the HNP protects mice against a lethal dose of histones through the inhibition of platelet aggregation and migration into the lungs. In vivo administration in murine sepsis model mice results in near complete survival. These results establish the potential for synthetic, nonbiological polymer hydrogel sequestrants as a new intervention strategy for sepsis therapy and adds to our understanding of the importance of histones to this condition.


Efficacy of alpha-Adrenergic Receptor Blockers in the Treatment of Male Lower Urinary Tract Symptoms.
Male lower urinary tract symptoms (LUTS) are one of the most common causes for a consultation with a health care provider, and one of the most common causes of male LUTS is benign prostatic hyperplasia (BPH). In recent decades, medical therapy has established itself as viable and cost effective for the majority of men. For the treatment of male LUTS in the United States, the 5 currently available alpha-adrenergic receptor blockers are alfuzosin, doxazosin, silodosin, terazosin, and tamsulosin. alpha-Blockers remain one of the mainstays in the treatment of male LUTS and clinical BPH. They exhibit an early onset of efficacy (within less than 1 week) with regard to both symptoms and flow rate improvement, maintain such improvements in open-label and controlled trials for up to 5 years, and have been shown to prevent symptomatic progression.


Quality of life after sipuleucel-T therapy: results from a randomized, double-blind study in patients with androgen-dependent prostate cancer.
To collect and analyze quality-of-life (QOL) data from PROvenge Treatment and Early Cancer Treatment trial (PROTECT, NCT00779402), a phase III, randomized controlled trial of sipuleucel-T in patients with asymptomatic androgen-dependent prostate cancer. Patients experiencing prostate-specific antigen relapse after radical prostatectomy entered a 3- to 4-month run-in phase of androgen-deprivation therapy (ADT), followed by 2:1 randomization to sipuleucel-T or control. QOL was assessed throughout the run-in and 26-week post-randomization phases using the Brief Fatigue Inventory (BFI), Linear Analog Self-Assessment (LASA) scale, Global Rating of Change (GRoC) scale, and an elicited symptoms list. One hundred seventy-six patients were randomized into 2 groups, the sipuleucel-T group (n = 117) or the control group (n = 59). The sample provided 80% power to detect a difference in fatigue interference score between treatment arms of 0.9 points. QOL declined predictably during ADT. At week 26, 26.2% of sipuleucel-T-treated patients and 21.6% of control-treated patients (P = .68) reported fatigue in the previous week, and the mean score for fatigue interference in the past 24 hours was 0.9 for both arms (P = .88). Results were comparable for usual fatigue (P = .91) and worst fatigue (P >.99). Mean LASA scores decreased in both groups (P = .26). The proportion of patients reporting better overall QOL on GRoC was similar (P = .62). There is no clinically significant negative impact on QOL after sipuleucel-T treatment compared with control after a period of ADT in patients with asymptomatic androgen-dependent prostate cancer.


Molecular basis of an autoantibody-associated restriction fragment length polymorphism that confers susceptibility to autoimmune diseases.
Recently, combined serological and molecular studies of autoantibodies have revealed that these antibodies play an important role in the normal function of the immune system and in the development of the B cell repertoire. Accordingly, we hypothesized that a homozygous deletion of a critical autoantibody-associated Ig variable (V) gene may alter the immune system and thus predispose the host to autoimmune disorders. Initial experiments revealed several restriction fragment length polymorphisms (RFLP) of the Humhv3005 gene, that is likely to encode heavy chains of rheumatoid factors, and the closely related 1.9III gene. By probing EcoR1-digested DNA with the Humhv3005/P1 probe, we found that one of the four major hybridizing bands was missing in approximately 20% of patients with either rheumatoid arthritis or systemic lupus erythematosus, but only 2% of normal subjects. To delineate the genetic basis of this polymorphism, we have now employed the PCR to amplify and analyze hv3005, 1.9III, and homologous genes in individuals with characteristic RFLP genotypes. Our results indicate that the human Vh gene repertoire contains several hv3005- and 1.9III-like genes, and that a complete deletion of the hv3005-like genes is relatively restricted to a subset of autoimmune patients. These findings provide initial evidence for deletion of developmentally regulated autoreactive V genes in autoimmune diseases.


Prognostic implication of glycolysis related gene signature in non-small cell lung cancer.
Abnormal glycolysis is one of the hallmarks of cancer and plays an important role in its development. This study was devoted to identify glycolysis related genes as prognostic biomarkers for non-small cell lung cancer (NSCLC). The mRNA expression profile and clinical follow-up data were obtained using The Cancer Genome Atlas (TCGA) database. The validation set was obtained by bootstrap method of random repeated sampling. A total of 200 glycolysis-related genes were obtained from Gene Set Enrichment Analysis (GSEA) and 46 genes were significantly associated with overall survival (OS). Five genes (PKP2, LDHA, HMMR, COL5A1 and B3GNT3) were eventually identified to calculate risk score of NSCLC patients. The univariate and multivariate Cox regression analysis indicated that the risk score was an independent prognostic factor (training set: HR=2.126, 95% CI [1.605, 2.815], p<0.001; validation set: HR=2.298, 95%CI [1.450, 3.640], p<0.001). Patients assigned to the high-risk group were associated with poor OS compared with patients in the low-risk group (training set: P=7.946e-06; validation set: P=6.368e-07). Receiver operating characteristic (ROC) curve and stratification analysis also demonstrated the potential prognostic performance. In conclusion, we constructed a novel glycolysis related risk signature which might contribute to predicting the prognosis of NSCLC.


Effect of weaning period on nutritional status of children.
To determine the effect of age of introduction and the type of weaning foods used, on the nutritional status of children under three years of age. Cross-sectional, community-based study. The study was conducted in rural areas of District Malir in Karachi, Pakistan, comprising of 900 villages spread over 1200 square kilometer, from November 2000 to April 2001. Three hundred and fifty-nine children under 3 years were included in the study. Based on the age of commencement of weaning, the children were categorized as children weaned at appropriate and inappropriate age. For the type of weaning foods, the children were grouped into two categories based on the nature of food items. Children for whom weaning was started at the recommended age of 4-6 months were found to be significantly less malnourished on anthropometric measurements as compared to the other set in which weaning was started at an earlier or later age. Significant statistical difference was noted in the children weaned on nutritious home-made items compared to children having commercial foods. Malnutrition is of multi-faceted origin beyond only poverty and food shortage. Inappropriate feeding practices can result in growth faltering as noted in the present study. Programmes should be launched for education of mothers regarding weaning practices.


The influence of psychosocial characteristics and race/ethnicity on the use, duration, and success of antiretroviral therapy.
Expanded access to antiretroviral therapy (ART) has produced dramatic reductions in HIV-associated morbidity and mortality. Disparities in access to and benefit from ART have been observed by race, gender, and mental health status, however. From 2001 to 2002, 611 HIV-positive patients were consecutively recruited from 5 southeastern US states and followed for 3 years. We evaluated demographic and psychosocial predictors of probability of receiving ART among all those eligible for ART (on ART, CD4 <350 cells/mm3 or viral load [VL] >55,000 copies/mL in the year preceding enrollment), time to first ART discontinuation among those on ART, and time to VL >400 copies/mL among those on ART with VL <400 copies/mL at enrollment. Of 611 participants, 474 consented to medical record abstraction and had known ART status at enrollment; 81% (385 of 474) of all participants and 89% (385 of 435) of ART-eligible participants were receiving ART at enrollment. In multivariable analyses, ART receipt was associated with greater age (adjusted odds ratio = 1.92 per 10 years, 95% confidence interval: 1.23 to 3.01), fewer recent stressful life events (odds ratio = 0.68, 95% confidence interval: 0.51 to 0.92), less alcohol use (odds ratio = 0.64, 95% confidence interval: 0.46 to 0.90), and greater perceived self-efficacy (OR = 2.82, 95% confidence interval: 1.41 to 5.62). No psychosocial characteristics were associated with ART discontinuation or virologic failure. No racial/ethnic or gender disparities were observed in ART receipt; however, minority racial/ethnic groups were faster to discontinue ART (adjusted hazard ratio = 2.44, 95% confidence interval: 1.33 to 4.49) and experience virologic failure (adjusted hazard ratio = 2.01, 95% confidence interval: 1.09 to 3.71). Patients with unfavorable psychosocial profiles were less likely to be on ART, perhaps attributable to providers' or patients' expectations of readiness. Psychosocial characteristics were not associated with ART discontinuation or virologic failure, however, possibly reflecting the selection process involved in who initiates ART. Racial disparities in ART discontinuation and virologic failure merit further attention.


Macrophage subsets harbouring Leishmania donovani in spleens of infected BALB/c mice: localization and characterization.
The purpose of the current study was to characterize parasite-containing cells located in spleens of BALB/c mice infected with Leishmania donovani. In particular, expression of MHC class II molecules by these cells was examined to determine whether they could potentially act as cells capable of immunostimulating Leishmania-reactive CD4+ T lymphocytes. To this end, an immunohistological analysis of spleens taken at various time points after infection was undertaken. Using this approach, we observed, in the red pulp, the formation of small cellular infliltrates containing heavily infected macrophages that could be stained with the monoclonal antibodies MOMA-2 and FA/11. All of them expressed high levels of MHC class II molecules. Parasites were also detected in the white pulp, especially in MOMA-2+, FA/11+ and MHC class II+ macrophages of the periarteriolar lymphocyte sheath and in MOMA-2+ marginal zone macrophages. Infected cells were further characterized by fluorescence microscopy after their enrichment by adherence. All infected mononuclear cells recovered by this procedure could be stained with MOMA-2 and FA/11 and thus very probably belonged to the mononuclear phagocyte lineage. Furthermore, all of them strongly expressed both MHC class II as well as H-2M molecules, regardless of the time points after infection. Analysis of the parasitophorous vacuoles (PV) by confocal microscopy showed that these compartments were surrounded by a membrane enriched in lysosomal glycoproteins lamp-1 and lamp-2, in macrosialin (a membrane protein of prelysosomes recognized by FA/11) and in MOMA-2 antigen. About 80% of the PV also had MHC class II and H-2M molecules on their membrane. Altogether, these data indicate that in the spleens of L. donovani-infected mice, a high percentage of amastigotes are located in macrophages expressing MHC class II molecules and that they live in PV exhibiting properties similar to those of PV detected in mouse bone marrow-derived macrophages exposed to a low dose of interferon gamma (IFN-gamma) and infected in vitro.


Prediagnostic Plasma Bile Acid Levels and Colon Cancer Risk: A Prospective Study.
Bile acids have been proposed to promote colon carcinogenesis. However, there are limited prospective data on circulating bile acid levels and colon cancer risk in humans. Associations between prediagnostic plasma levels of 17 primary, secondary, and tertiary bile acid metabolites (conjugated and unconjugated) and colon cancer risk were evaluated in a nested case-control study within the European Prospective Investigation into Cancer and Nutrition (EPIC) cohort. Bile acid levels were quantified by tandem mass spectrometry in samples from 569 incident colon cancer cases and 569 matched controls. Multivariable logistic regression analyses were used to estimate odds ratios (ORs) for colon cancer risk across quartiles of bile acid concentrations. Positive associations were observed between colon cancer risk and plasma levels of seven conjugated bile acid metabolites: the primary bile acids glycocholic acid (ORquartile 4 vs quartile 1= 2.22, 95% confidence interval [CI] = 1.52 to 3.26), taurocholic acid (OR = 1.78, 95% CI = 1.23 to 2.58), glycochenodeoxycholic acid (OR = 1.68, 95% CI = 1.13 to 2.48), taurochenodeoxycholic acid (OR = 1.62, 95% CI = 1.11 to 2.36), and glycohyocholic acid (OR = 1.65, 95% CI = 1.13 to 2.40), and the secondary bile acids glycodeoxycholic acid (OR = 1.68, 95% CI = 1.12 to 2.54) and taurodeoxycholic acid (OR = 1.54, 95% CI = 1.02 to 2.31). By contrast, unconjugated bile acids and tertiary bile acids were not associated with risk. This prospective study showed that prediagnostic levels of certain conjugated primary and secondary bile acids were positively associated with risk of colon cancer. Our findings support experimental data to suggest that a high bile acid load is colon cancer promotive.


DL-a-Monofluoromethylputrescine is a potent irreversible inhibitor of Escherichia coli ornithine decarboxylase.
DL-alpha-Monofluoromethylputrescine (compound R.M.I. 71864) is an enzyme-activated irreversible inhibitor of the biosynthetic enzyme ornithine decarboxylase from Escherichia coli. This compound, however, has much less effect in vitro on ornithine decarboxylase obtained from Pseudomonas aeruginosa. These findings are in contrast with those previously found with the substrate analogue DL-alpha-difluoromethylornithine (compound R.M.I. 71782). The K1 of the DL-alpha-monofluoromethylputrescine for the E. coli ornithine decarboxylase is 110 microM, and the half-life (t1/2) calculated for an infinite concentration of inhibitor is 2.1 min. When DL-alpha-monofluoromethylputrescine is used in combination with DL-alpha-difluoromethylarginine (R.M.I. 71897), an irreversible inhibitor of arginine decarboxylase, in vivo in E. coli, both decarboxylase activities are inhibited (greater than 95%) but putrescine levels are only decreased to about one-third of control values and spermidine levels are slightly increased.


Adverse health effects of particulate air pollution: modification by air conditioning.
The short-term effects of particulate matter (PM) on mortality and morbidity differ by geographic location and season. Several hypotheses have been proposed for this variation, including different exposures with air conditioning (AC) versus open windows. Bayesian hierarchical modeling was used to explore whether AC prevalence modified day-to-day associations between PM10 and mortality, and between PM2.5 and cardiovascular or respiratory hospitalizations, for those 65 years and older. We considered yearly, summer-only, and winter-only effect estimates and 2 types of AC (central and window units). Communities with higher AC prevalence had lower PM effects. Associations were observed for cardiovascular hospitalizations and central AC. Each additional 20% of households with central AC was associated with a 43% decrease in PM2.5 effects on cardiovascular hospitalization. Central AC prevalence explained 17% of between-community variability in PM2.5 effect estimates for cardiovascular hospitalizations. Higher AC prevalence was associated with lower health effect estimates for PM.


Sex differences in complaints and diagnoses.
This paper examines male-female differences in complaints and diagnoses for ambulatory care visits. Data are from the 1973 National Ambulatory Medical Care Survey, a national probability survey of visits to office-based physicians. The results suggest that: (1) Men are often unaware of serious health problems, they delay seeking diagnosis and care for symptoms, and they hesitate to admit symptoms and known health problems when they do visit a physician. (2) Women appear to have a more diffuse view of illness. They often report both mental and physical symptoms, and their physical symptoms "radiate" throughout the body rather than remain localized. (3) Both sexes confuse reproductive, digestive, and urinary symptoms because the body systems overlap. (4) Some sex differences in diagnoses for a particular symptom reflect real morbidity differences. (5) There is little evidence that women and men differ in their perception, interpretation, and description of physical symptoms. (6) The evidence for sex bias in physicians' diagnoses is scant.


Gastric coinfection with thiopeptide-positive Cutibacterium acnes decreases FOXM1 and pro-inflammatory biomarker expression in a murine model of Helicobacter pylori-induced gastric cancer.
H. pylori infects half of the world population and is the leading cause of gastric cancer. We previously demonstrated that gastric cancer risk is associated with gastric microbiota. Specifically, gastric urease-positive Staphylococcus epidermidis and Streptococcus salivarius had contrasting effects on H. pylori-associated gastric pathology and immune responses in germ-free INS-GAS mice. As gastritis progresses to gastric cancer, the oncogenic transcription factor Foxm1 becomes increasingly expressed. In this study, we evaluated the gastric commensal C. acnes, certain strains of which produce thiopeptides that directly inhibit FOXM1. Thiopeptide-positive C. acnes was isolated from Nicaraguan patient gastric biopsies and inoculated into germ-free INS-GAS mice with H. pylori. We, therefore, asked whether coinfection with C. acnes expressing thiopeptide and H. pylori would decrease gastric Foxm1 expression and pro-inflammatory cytokine mRNA and protein levels. Our study supports the growing literature that specific non-H. pylori gastric bacteria affect inflammatory and cancer biomarkers in H. pylori pathogenesis.


Adjuvant chemotherapy with vincristine, cyclophosphamide, and doxorubicin after radiotherapy in local-regional nasopharyngeal cancer: results of a 4-year multicenter randomized study.
To evaluate the effect of adjuvant chemotherapy in patients with local-regional nasopharyngeal carcinoma (NPC) (squamous or undifferentiated) in complete remission at the end of curative radiotherapy (RT) 229 patients were randomized from 1979 to 1983 in a multicenter study to no further therapy (116 patients) or a combination of vincristine, cyclophosphamide, and Adriamycin (doxorubicin; Adria Laboratories, Columbus, OH) (VCA) for six monthly cycles (113 patients). The RT and RT + VCA groups were well balanced for median age (50 v 49 years), histology (undifferentiated carcinoma, 73% v 70%), tumor extent (tumor limited to nasopharynx, 57% v 57%), and nodal extent (negative nodes 26% v 24%, nodes in the lower cervical levels, 17% v 16%). RT was delivered to the nasopharynx, the base of the skull, and bilateral cervical nodes using a split course technique over 10 weeks up to the dose of 60 to 70 Gy in involved sites and 50 Gy to negative nodes. Response to RT was evaluated within 65 days post-RT treatment. Analysis at 48 months did not show significant difference between the two treatment groups in terms of relapse-free survival (RT, 55.8%, RT + VCA, 57.7%, P = .45) and overall survival (RT, 67.3%, RT + VCA, 58.5%, P = .13). The pattern of relapse was similar in the two treatment arms. Distant metastases were the cause of treatment failure in about 50% of relapsing patients. Although the results of the present study did not show any benefit from VCA administered after curative RT, combined systemic chemotherapy should be further explored due to the high incidence of local and distant failure after intensive RT.


Amygdalin Attenuates Airway Epithelium Apoptosis, Inflammation, and Epithelial-Mesenchymal Transition through Restraining the TLR4/NF-κB Signaling Pathway on LPS-Treated BEAS-2B Bronchial Epithelial Cells.
Cough-variant asthma (CVA) is a special type of asthma, solely manifesting with coughing. Studies suggest that airway inflammation is associated with CVA pathogenesis. Amygdalin is found to have an anti-inflammatory potential, while how it affects CVA remains unexplored. Cytotoxicity delivered by various concentrations of LPS and amygdalin on BEAS-2B cells was determined by Cell Counting Kit-8 assay. CVA in vitro models were established via LPS exposure on BEAS-2B cells which underwent amygdalin pretreatment. Cell apoptosis was determined by flow cytometry. Production of tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-8, and mucin 5AC (MUC5AC) in BEAS-2B cells was measured by ELISA and qRT-PCR. Expressions of TLR4, E-cadherin, N-cadherin, α-smooth muscle actin (SMA), vimentin, phosphorylated-p65 (p-p65), p65, phosphorylated-IκBα (p-IκBα), and IκBα in BEAS-2B cells were measured by qRT-PCR or Western blot. LPS and high concentrations of amygdalin (over 600 μg/mL) decreased BEAS-2B cell toxicity. Exposure to LPS inhibited toxicity, enhanced apoptosis; and promoted production of TNF-α, IL-6, IL-8, and MUC5AC, increased the levels of N-Cadherin, α-SMA, vimentin, p-p65, and p-IκBα, and decreased the levels of E-cadherin and IκBα in BEAS-2B cells. Amygdalin pretreatment counteracted the effects of LPS on BEAS-2B cells. Overexpressing TLR4 reversed amygdalin-exerted effects in LPS-exposed BEAS-2B cells. Amygdalin attenuated airway epithelium apoptosis, inflammation and epithelial-mesenchymal transition through restraining the TLR4/NF-κB signaling pathway in CVA.


Glucose metabolism in Goto-Kakizaki rat islets.
Islets from Goto-Kakizaki (GK) rats from our colony, despite marked impairment of glucose-induced insulin release, used glucose and produced CO2 at a rate 3 times that of islets from control Wistar rats. Almost all glucose used was accounted for in CO2 and lactate production. The percentages of glucose carbon used collected in CO2 and lactate were similar for control and GK islets. GK islets also oxidized 40% more acetate and leucine to CO2 than did control islets. The fraction of carbon leaving the Krebs cycle relative to CO2 production was the same in GK and control islets. The capacities of mitochondria from GK islets to generate ATP from glutamate and malate were similar and that to generate ATP from succinate and rotenone was somewhat less from GK islets. The reason for the enhanced utilization of substrates by islets of the GK rat is not apparent. In conclusion, there is no decrease in islet glucose utilization, glucose oxidation, Krebs cycle function, or the electron transport system evident from these measurements to explain the impaired insulin release in islets from GK rats.


A Clinically Evaluated Interferometric Continuous-Wave Radar System for the Contactless Measurement of Human Vital Parameters.
Vital parameters are key indicators for the assessment of health. Conventional methods rely on direct contact with the patients' skin and can hence cause discomfort and reduce autonomy. This article presents a bistatic 24 GHz radar system based on an interferometric six-port architecture and features a precision of 1 µm in distance measurements. Placed at a distance of 40 cm in front of the human chest, it detects vibrations containing respiratory movements, pulse waves and heart sounds. For the extraction of the respiration rate, time-domain approaches like autocorrelation, peaksearch and zero crossing rate are compared to the Fourier transform, while template matching and a hidden semi-Markov model are utilized for the detection of the heart rate from sphygmograms and heart sounds. A medical study with 30 healthy volunteers was conducted to collect 5.5 h of data, where impedance cardiogram and electrocardiogram were used as gold standard for synchronously recording respiration and heart rate, respectively. A low root mean square error for the breathing rate (0.828 BrPM) and a high overall F1 score for heartbeat detection (93.14%) could be achieved using the proposed radar system and signal processing.


Reactivity of Biliatresone, a Natural Biliary Toxin, with Glutathione, Histamine, and Amino Acids.
In our previous work, we identified a natural toxin, biliatresone, from Dysphania glomulifera and D. littoralis, endemic plants associated with outbreaks of biliary atresia in Australian neonatal livestock. Biliatresone is a very rare isoflavonoid with an α-methylene ketone between two phenyls, 1,2-diaryl-2-propenone, along with methylenedioxy, dimethoxyl, and hydroxyl functional groups, that causes extrahepatic biliary toxicity in zebrafish. The toxic core of biliatresone is a methylene in the α-position relative to the ketone of 1,2-diaryl-2-propenone that serves as an electrophilic Michael acceptor. The α-methylene of biliatresone spontaneously conjugated with water and methanol (MeOH), respectively, via Michael addition in a reverse phase high-performance liquid chromatography (RP-HPLC) analysis. We here report the reactivity of biliatresone toward glutathione (GSH), several amino acids, and other thiol- or imidazole-containing biomolecules. LC-MS and HPLC analysis of the conjugation reaction showed the reactivity of biliatresone to be in the order histidine > N-acetyl-d-cysteine (D-NAC) = N-acetyl-l-cysteine (L-NAC) > histamine > glutathione ≥ cysteine ≫ glycine > glutamate > phenylalanine, while serine and adenine had no reactivity due to intramolecular hydrogen bonding in the protic solvents. The reactivity of ethyl vinyl ketone (EVK, 1-penten-3-one), an example of a highly reactive α,ß-unsaturated ketone, toward GSH gave a 6.7-fold lower reaction rate constant than that of biliatresone. The reaction rate constant of synthetic 1,2-diaryl-2-propen-1-one (DP), a core structure of the toxic molecule, was 10-fold and 1.5-fold weaker in potency compared to the reaction rate constants of biliatresone and EVK, respectively. These results demostrated that the methylenedioxy, dimethoxyl, and hydroxyl functional groups of biliatresone contribute to the stronger reactivity of the Michael acceptor α-methylene ketone toward nucleophiles compared to that of DP and EVK.


Structures and mechanisms of Nudix hydrolases.
Nudix hydrolases catalyze the hydrolysis of nucleoside diphosphates linked to other moieties, X, and contain the sequence motif or Nudix box, GX(5)EX(7)REUXEEXGU. The mechanisms of Nudix hydrolases are highly diverse in the position on the substrate at which nucleophilic substitution occurs, and in the number of required divalent cations. While most proceed by associative nucleophilic substitutions by water at specific internal phosphorus atoms of a diphosphate or polyphosphate chain, members of the GDP-mannose hydrolase sub-family catalyze dissociative nucleophilic substitutions, by water, at carbon. The site of substitution is likely determined by the positions of the general base and the entering water. The rate accelerations or catalytic powers of Nudix hydrolases range from 10(9)- to 10(12)-fold. The reactions are accelerated 10(3)-10(5)-fold by general base catalysis by a glutamate residue within, or beyond the Nudix box, or by a histidine beyond the Nudix box. Lewis acid catalysis, which contributes 10(3)-10(5)-fold to the rate acceleration, is provided by one, two, or three divalent cations. One divalent cation is coordinated by two or three conserved residues of the Nudix box, the initial glycine and one or two glutamate residues, together with a remote glutamate or glutamine ligand from beyond the Nudix box. Some Nudix enzymes require one (MutT) or two additional divalent cations (Ap(4)AP), to neutralize the charge of the polyphosphate chain, to help orient the attacking hydroxide or oxide nucleophile, and/or to facilitate the departure of the anionic leaving group. Additional catalysis (10-10(3)-fold) is provided by the cationic side chains of lysine and arginine residues and by H-bond donation by tyrosine residues, to orient the general base, or to promote the departure of the leaving group. The overall rate accelerations can be explained by both independent and cooperative effects of these catalytic components.


p53, p16 and Fhit proteins expressions in chronic esophagitis and Chagas disease.
Models have suggested esophageal carcinogenesis can result from the alteration of sequences, leading to esophagitis, atrophy, dysplasia, carcinoma in situ and invasive carcinoma. While numerous genetic alterations have been reported in esophageal carcinogenesis, studies of benign lesions with precancerous potential are scarce. Immunohistochemistry was performed for p53, p16 and Fhit proteins in the esophageal mucosa from patients with Chagas disease (CD), chagasic megaesophagus (CM), chronic esophagitis (CE), esophageal squamous cell carcinoma (ESCC) and in normal mucosa (NM). The proportion of p53-positive cases increased progressively according to the severity of the pathology CD (7.7%), CM (26.1%), CE (522%) and ESCC (100%). However, p16 and Fhit did not show any statistically significant differences among the groups. p53 overexpression is involved in the initial steps of esophageal carcinogenesis, supporting further evaluation of its utility as a marker in precursor lesions, conversely, losses of Fhit and p16 expression may not be significant.


Self-Reported Sedative Drug Use Among Students Attending at University of Gondar, Gondar, Northwest, Ethiopia: A Cross-Sectional Study.
Medical students experience significant psychological stress and are therefore at higher risk of using sedatives. There are currently no studies describing the prevalence of sedative drug use among medical students in Ethiopia. This study aimed to assess the prevalence and associated factors of self-reported sedative drug use among medical students attending the College of Medicine and Health Science (CMHS) students at the University of Gondar (UoG). A prospective cross-sectional study was conducted from May to July 2018 in CMHS at UoG. Data were collected using a pre-tested self-administered standard questionnaire. Data were collected, entered into a computer using Epi Info 7 software, and analyzed using SPSS version 20. Frequency, mean, and standard deviation were used to describe descriptive statistics, and binary and multiple logistic regression analyses were used to assess the association between different variables and sedative drug use; P <0.05 was used to declare association. Of the 422 students who returned questionnaires, 26 (6.2%) participants were reported sedative drug use at some time since enrollment. Of these, 61.54% participants used antihistamine drugs. Smoking status (AOR (95% CI), 0.046 (0.009-0.241) P = 0.0001), stimulant use (AOR (95% CI), 0.220 (0.062-0.780) P = 0.019), sleeping hour (AOR (95% CI), 9.931 (4.155-14.785) P = 0.001) and sleep disorder (AOR (95% CI), 0.149 (0.033-0.680) P = 0.014) were significantly associated with sedative drug use. Self-reported sedative drug use among medical students at the University of Gondar is relatively low, and antihistamines are the most commonly used drugs. Smoking, stimulant use, sleeping hour, and the presence of sleep disorders were associated with sedative drug use.


HPLC-MS/MS method for quantitative determination of the novel dual inhibitor of FGF and VEGF receptors E-3810 in tumor tissues from xenograft mice and human biopsies.
We developed and validated a high-performance liquid chromatography-tandem mass spectrometry analytical method to measure E-3810, a novel dual inhibitor of fibroblast growth factor receptor 1 and vascular endothelial growth factor receptor 1-3 in tissues and determined the drug concentration in a biopsy of human breast cancer for the first time. The method is a modification of our previous one in plasma to study the clinical pharmacokinetics of the drug during the phase I/II trial. In view of the changes in matrix, we applied a partial validation protocol to determine recovery, sensitivity, range of linearity, precision, accuracy and stability of the method over three runs in a mouse tumor tissue and liver. The recovery of E-3810 from liver or tumor homogenate was >69%, and the lower limit of quantification was 5 ng/ml. The method was linear in the concentration range 5.0-500.0 ng/ml, as demonstrated by a determination coefficient R(2) ≥ 0.9955. The range of the calibration curve was appropriate for the analysis, as demonstrated by the accuracy, which was between 91.4% and 106.7%. Interday precision and accuracy on quality control samples at 9, 30 and 300 ng/ml were 3.1-11.2% and 98.3-111.4%, respectively. The assay was applied successfully to determine the intratumor concentration of E-3810 in different mouse xenograft tumor models and in a biopsy of a patient with breast cancer included in the phase I/II trial of the drug. In mouse tumors, the concentrations of E-3810 were higher than necessary to exert antitumor activity in vitro (1 µM). Even more of interest was the result obtained in a human biopsy of few milligrams, where E-3810 reached 4.9 µg/g (11 µM).


Subcellular targeting and biosynthesis of cyclotides in plant cells.
The cyclotide kalata B1 is found in the leaves of Oldenlandia affinis and is a potent insecticidal and nematocidal molecule. This peptide is cleaved from a precursor protein, Oak1, and ligation of the N- and C-termini occurs to form a continuous peptide backbone. The subcellular location of the excision and cyclization reactions is unknown, and there is debate as to which enzyme catalyzes the event. To determine where in the plant cell Oak1 is processed, we prepared constructs encoding GFP (green fluorescent protein) linked to the cyclotide precursor Oak1. The GFP constructs were transiently expressed in the leaves of Nicotiana benthamiana, and GFP fluorescence was observed in living cells using confocal microscopy. A Fei Mao (FM) styryl dye was infiltrated into whole leaves that were still growing and expressing GFP constructs, enabling the plasma membrane and the tonoplast to be highlighted for visualization of the vacuole in living cells. The full length Oak1 precursor directed GFP to the vacuole, suggesting that excision and cyclization of the cyclotide domain occurs in the vacuole where the cyclotides are then stored. The N-terminal propeptide and N-terminal repeat of Oak1 were both sufficient to target GFP to the vacuole, although the C-terminal propeptide, which is essential for cyclization, was not a targeting signal. The vacuolar location of cyclotides supports our hypothesis that the vacuolar processing enzyme, asparaginyl endoproteinase, has a pivotal role in excision and cyclization from cyclotide precursors.


Algorithmic handwriting analysis of Judah's military correspondence sheds light on composition of biblical texts.
The relationship between the expansion of literacy in Judah and composition of biblical texts has attracted scholarly attention for over a century. Information on this issue can be deduced from Hebrew inscriptions from the final phase of the first Temple period. We report our investigation of 16 inscriptions from the Judahite desert fortress of Arad, dated ca 600 BCE-the eve of Nebuchadnezzar's destruction of Jerusalem. The inquiry is based on new methods for image processing and document analysis, as well as machine learning algorithms. These techniques enable identification of the minimal number of authors in a given group of inscriptions. Our algorithmic analysis, complemented by the textual information, reveals a minimum of six authors within the examined inscriptions. The results indicate that in this remote fort literacy had spread throughout the military hierarchy, down to the quartermaster and probably even below that rank. This implies that an educational infrastructure that could support the composition of literary texts in Judah already existed before the destruction of the first Temple. A similar level of literacy in this area is attested again only 400 y later, ca 200 BCE.


APP, PSEN1, and PSEN2 mutations in early-onset Alzheimer disease: A genetic screening study of familial and sporadic cases.
Amyloid protein precursor (APP), presenilin-1 (PSEN1), and presenilin-2 (PSEN2) mutations cause autosomal dominant forms of early-onset Alzheimer disease (AD-EOAD). Although these genes were identified in the 1990s, variant classification remains a challenge, highlighting the need to colligate mutations from large series. We report here a novel update (2012-2016) of the genetic screening of the large AD-EOAD series ascertained across 28 French hospitals from 1993 onwards, bringing the total number of families with identified mutations to n = 170. Families were included when at least two first-degree relatives suffered from early-onset Alzheimer disease (EOAD) with an age of onset (AOO) ≤65 y in two generations. Furthermore, we also screened 129 sporadic cases of Alzheimer disease with an AOO below age 51 (44% males, mean AOO = 45 ± 2 y). APP, PSEN1, or PSEN2 mutations were identified in 53 novel AD-EOAD families. Of the 129 sporadic cases screened, 17 carried a PSEN1 mutation and 1 carried an APP duplication (13%). Parental DNA was available for 10 sporadic mutation carriers, allowing us to show that the mutation had occurred de novo in each case. Thirteen mutations (12 in PSEN1 and 1 in PSEN2) identified either in familial or in sporadic cases were previously unreported. Of the 53 mutation carriers with available cerebrospinal fluid (CSF) biomarkers, 46 (87%) had all three CSF biomarkers-total tau protein (Tau), phospho-tau protein (P-Tau), and amyloid β (Aβ)42-in abnormal ranges. No mutation carrier had the three biomarkers in normal ranges. One limitation of this study is the absence of functional assessment of the possibly and probably pathogenic variants, which should help their classification. Our findings suggest that a nonnegligible fraction of PSEN1 mutations occurs de novo, which is of high importance for genetic counseling, as PSEN1 mutational screening is currently performed in familial cases only. Among the 90 distinct mutations found in the whole sample of families and isolated cases, definite pathogenicity is currently established for only 77%, emphasizing the need to pursue the effort to classify variants.


A propagated wave of MPF activation accompanies surface contraction waves at first mitosis in Xenopus.
During the period of mitosis, two surface contraction waves (SCWs) progress from the animal to vegetal poles of the Xenopus egg. It has been shown that these SCWs occur in parallel with the activation of MPF and with its subsequent inactivation in the animal and vegetal hemispheres, suggesting that they are responses to propagated waves of MPF activity across the egg. We have analysed the mechanism of MPF regulation in different regions of the egg in detail in relation to SCW progression. The distributions of histone HI kinase activity and of Cdc2 and cyclin B (the catalytic and regulatory subunits of MPF) were followed by dissection of intact eggs following freezing and in cultured fragments separated by ligation. Cdc2 was found to be distributed evenly throughout the egg cytoplasm. Loss of phosphorylated (inactive) forms of Cdc2 coincided spatially with the wave of MPF activation, while cyclin B2 accumulation occurred in parallel in animal and vegetal regions. In ligated vegetal pole fragments no MPF activation or Cdc2 dephosphorylation were detectable. A wave of cyclin B destruction that occurred in concert with the second SCW was also blocked. Taken together these results indicate that the triggering mechanism for MPF activation requires components specific to the animal cytoplasm, acting via Cdc2 dephosphorylation, and that MPF activation subsequently propagates autocatalytically across the egg. SCW progression in the vegetal hemisphere was followed directly by time-lapse videomicroscopy of subcortical mitochondrial islands. The first SCW traversed the vegetal pole at the time of MPF activation in this region. Like MPF activation and inactivation, SCWs were blocked in the vegetal region by ligation. These observations reinforce the hypothesis that the first SCW is a direct consequence of the MPF activation wave. It may reflect depolymerisation of the subcortical microtubule network since it coincided exactly with the arrest of the microtubule-dependent movement of 'cortical rotation' and was related in direction in most eggs. The cyclin B destruction wave and associated cortical contraction of the second SCW may be localised downstream consequences of the MPF activation wave, or they may propagate independently from the animal cytoplasm.


Germ Cell-less Promotes Centrosome Segregation to Induce Germ Cell Formation.
The primordial germ cells (PGCs) specified during embryogenesis serve as progenitors to the adult germline stem cells. In Drosophila, the proper specification and formation of PGCs require both centrosomes and germ plasm, which contains the germline determinants. Centrosomes are microtubule (MT)-organizing centers that ensure the faithful segregation of germ plasm into PGCs. To date, mechanisms that modulate centrosome behavior to engineer PGC development have remained elusive. Only one germ plasm component, Germ cell-less (Gcl), is known to play a role in PGC formation. Here, we show that Gcl engineers PGC formation by regulating centrosome dynamics. Loss of gcl leads to aberrant centrosome separation and elaboration of the astral MT network, resulting in inefficient germ plasm segregation and aborted PGC cellularization. Importantly, compromising centrosome separation alone is sufficient to mimic the gcl loss-of-function phenotypes. We conclude Gcl functions as a key regulator of centrosome separation required for proper PGC development.


Kisspeptin antagonist prevents RF9-induced reproductive changes in female rats.
The aim of this study was to determine the modulatory effects of peptide 234 (p234) (an antagonist of GPR54 receptors) on kisspeptin and RF9 (an RFamide-related peptide antagonist)-induced changes in reproductive functions and energy balance in female rats. Female Sprague-Dawley rats were weaned on postnatal day (pnd) 21. The animals were intracerebroventricularly cannulated under general anesthesia on pnd 23. Groups of female rats were injected with kisspeptin, RF9, p234, kisspeptin plus p234, or RF9 plus p234, daily. The experiments were ended on the day of first diestrus following pnd 60. Kisspeptin or RF9 alone advanced vaginal opening (VO), which was delayed by administration of kisspeptin antagonist alone. In the rats given kisspeptin plus p234 or RF9 plus p234, VO was not different from control rats. Kisspeptin and RF9 elicited significant elevations in circulating LH levels. Coadministrations of kisspeptin or RF9 with p234 decreased LH levels significantly. The use of p234 alone did not cause any significant change in LH secretion. Kisspeptin decreased both food intake and body weight while RF9 decreased only food intake without affecting body weight. The effects of kisspeptin on energy balance were also reversed by central administration of p234. In conclusion, kisspeptin antagonist, p234, modulates the effects of kisspeptin on reproductive functions and energy balance, whereas RF9 seems to exert only its effects on reproductive functions by means of GPR54 signaling in female rats.


Analysis of Fish-Consumption Benefits and Safety Knowledge in a Population-Based Sample of Polish Adolescents.
Inadequate fish consumption is common and may result from multiple reasons, especially in adolescents who are a population at particular risk of the negative consequences of not consuming the recommended amounts of fish. The aim of the study was to analyze the knowledge about fish-consumption benefits and safety in a population-based sample of Polish adolescents. The stratified random sampling was conducted within two stages: sampling of counties from all voivodeships in Poland (being the basic administrative units of Poland) and inviting secondary schools from the drawn counties to obtain a sample representative of all regions of Poland. The Computer-Assisted Web Interview (CAWI) method was applied to gather the data within the study, and a questionnaire concerning knowledge about fish-consumption benefits and safety with 20 true-false statements was applied. Among 1289 participants, the statement with the highest share of correct answers concerned fish being a source of protein (78.9%) and fish-derived fats being health promoting (77.0%). The statement receiving the least correct answers concerned the type of fatty acids found in fish (7.6%) and the risk of polychlorinated biphenyls (PCBs) in fish (20.5%). Participants who were female, older than 18, underweight, living in an urban environment, from a region far away from the sea and from comprehensive schools provided a higher share of correct answers than other subgroups (p < 0.05). Knowledge concerning fish-consumption benefits and safety among Polish adolescents is in many cases inadequate; thus, nutritional education is needed, especially among younger adolescents, those attending vocational schools, males and those living in a rural environment.


US hospice industry experienced considerable turbulence from changes in ownership, growth, and shift to for-profit status.
The US hospice industry, which provides palliative and supportive care to patients with terminal illness, has undergone substantial changes during the last decade. The magnitude of these changes has not been fully captured in previous studies or reports. In this longitudinal study of hospices active in Medicare during 1999-2009, we analyzed Provider of Services files to understand key shifts in the industry. We found evidence of substantial turbulence. One-fifth of Medicare-certified hospices active in 1999 had closed or withdrawn from the program by 2009, and more than 40 percent had experienced one or more changes in ownership. The most prominent trend was the shift in ownership type from nonprofit to for-profit ownership. Four out of five Medicare-certified hospices that entered the marketplace between 2000 and 2009 were for-profit. Hospices also became larger, as the proportion with 100 or more full-time employees doubled to 5 percent from 1999 to 2009. Although each of the Census regions had more hospices in 2009 than in 1999, the geographic distribution of hospices in the country changed, with proportionally more in the South and West. The impact of all of these changes on cost and quality of hospice care, as well as patient access, remains a critical area for future research.


Imaging tumor angiogenesis by use of combined near-infrared diffusive light and ultrasound.
A novel two-step reconstruction scheme using a combined near-infrared and ultrasound technique and its utility in imaging distributions of optical absorption and hemoglobin concentration of breast lesions are demonstrated. In the first-step image reconstruction, the entire tissue volume is segmented based on initial coregistered ultrasound measurements into lesion and background regions. Reconstruction is performed by use of a finer grid for lesion region and a coarse grid for the background tissue. As a result, the total number of voxels with unknown absorption can be maintained on the same order of total measurements, and the matrix with unknown total absorption distribution is appropriately scaled for inversion. In the second step, image reconstruction is refined by optimization of lesion parameters measured from ultrasound images. It is shown that detailed distributions of wavelength-dependent absorption and hemoglobin concentration of breast carcinoma can be obtained with the new reconstruction scheme.


Neural representation of object approach in a decision-making motor circuit.
Although behavior is ultimately guided by decision-making neurons and their associated networks, the mechanisms underlying neural decision-making in a behaviorally relevant context remain mostly elusive. To address this question, we analyzed goldfish escapes in response to distinct visual looming stimuli with high-speed video and compared them with electrophysiological responses of the Mauthner cell (M-cell), the threshold detector that initiates such behaviors. These looming stimuli evoke powerful and fast body-bend (C-start) escapes with response probabilities between 0.7 and 0.91 and mean latencies ranging from 142 to 716 ms. Chronic recordings showed that these C-starts are correlated with M-cell activity. Analysis of response latency as a function of the different optical parameters characterizing the stimuli suggests response threshold is closely correlated to a dynamically scaled function of angular retinal image size, (t), specifically kappa(t) = (t-delta x e(-beta(t-delta)), where the exponential term progressively reduces the weight of (t). Intracellular recordings show that looming stimuli typically evoked bursts of graded EPSPs with peak amplitudes up to 9 mV in the M-cell. The proposed scaling function kappa(t) predicts the slope of the depolarizing envelope of these EPSPs and the timing of the largest peak. An analysis of the firing rate of presynaptic inhibitory interneurons suggests the timing of the EPSP peak is shaped by an interaction of excitatory and inhibitory inputs to the M-cell and corresponds to the temporal window in which the probabilistic decision of whether or not to escape is reached.