The effects of a 12-week strength-training program on strength and functionality in women with fibromyalgia.
To determine whether women with fibromyalgia benefit from strength training. Randomized controlled trial. Testing was completed at the university and training was completed at a local community wellness facility. Twenty-nine women (age range, 18-54 y) with fibromyalgia participated. Subjects were randomly assigned to a control (n=14; wait-listed for exercise) or strength (n=15) group. After the first 4 weeks, 7 (47%) women dropped from the strength group. Subjects underwent 12 weeks of training on 11 exercises, 2 times a week, performing 1 set of 8 to 12 repetitions at 40% to 60% of their maximal lifts and were progressed to 60% to 80%. Subjects were measured for strength, functionality, tender point sensitivity, and fibromyalgia impact. The strength group significantly (P< or =.05) improved upper- (strength, 39+/-11 to 42+/-12 kg; control, 38+/-13 to 38+/-12 kg) and lower- (strength, 68+/-28 to 82+/-25 kg; control, 61+/-25 to 61+/-26 kg) body strength. Upper-body functionality measured by the Continuous-Scale Physical Functional Performance test improved significantly (strength, 44+/-11 to 50+/-16U; control, 51+/-11 to 49+/-13U) after training. Tender point sensitivity and fibromyalgia impact did not change. Strength training improved strength and some functionality in women with fibromyalgia. Interventions with resistance have important implications on independence and quality of life issues for women with fibromyalgia.


Vector-Borne Pathogens in Stray Cats in Eastern Germany (Thuringia).
Bacterial, protozoal, and viral vector-borne pathogens (VBPs) can cause infections in cats. There is little information on feline VBP prevalence in Germany. Stray cats are frequently exposed to vectors but receive no veterinary care. The aim of this study was to determine the prevalence of selected VBPs in stray cats. EDTA blood and serum samples were taken from apparently healthy stray cats during a spay/neuter campaign in the federal state of Thuringia. Overall, 11/50 (22%) and 32/50 (64%) cats tested positive for at least one VBP by direct and indirect detection methods, respectively. PCR testing of EDTA blood detected hemotropic Mycoplasma spp. in 12% of cats, Hepatozoon spp. in 10%, and Anaplasma phagocytophilum in 4%. PCR testing for Rickettsia spp. and piroplasms was negative. IFAT on serum samples showed 46% of cats had detectable antibodies for Bartonella spp., 30% for Rickettsia spp., and 16% for A. phagocytophilum. The cats were additionally tested for feline coronavirus, FIV, and FeLV to identify potential risk factors for pathogen contact and/or infections. No correlation between FIV and FeLV status and VBP positivity was detected. Anaplasma phagocytophilum, Rickettsia spp., and Bartonella spp. have zoonotic potential, and surveillance is recommended in the context of the One Health approach.


Genotyping of Toxoplasma gondii Strains from Goats in Jahrom District, Southern Iran.
Toxoplasma gondii is transmitted congenitally or acquired by consumption of food and water contaminated with cysts or oocysts. This study aimed at genotyping T. gondii strains from slaughtered goats in Jahrom. A total of 561 specimens (heart, diaphragm, and tongue) from 187 slaughtered goats were collected from Jahrom slaughterhouse. After DNA extraction, the T. gondii strains were genotyped by the nested PCR-RFLP based on GRA6 and 3', and 5' ends of the SAG2 gene. T. gondii infection was present in 18.2% of cases. Among the examined organs, the diaphragm was more disposed to the infection (10.2%). Furthermore, infection rates of the heart and tongue were 8.6% and 3.7%, respectively. Concurrent infection in the heart and diaphragm, tongue and diaphragm, and heart and tongue were 3.2%, 0.5%, and 0.5%, respectively. In genotyping experiments, genotype I was the most frequent genotype of T. gondii (58.8%), followed by type II (23.5%), type III (11.8%), and a combination of type I and II (5.9%). The results of this study showed the presence of different genotypes of T. gondii in goats including three major and mixed genotypes. These results can be useful in toxoplasmosis control and prevention.


Precise Network Modeling of Systems Genetics Data Using the Bayesian Network Webserver.
The Bayesian Network Webserver (BNW, http://compbio.uthsc.edu/BNW ) is an integrated platform for Bayesian network modeling of biological datasets. It provides a web-based network modeling environment that seamlessly integrates advanced algorithms for probabilistic causal modeling and reasoning with Bayesian networks. BNW is designed for precise modeling of relatively small networks that contain less than 20 nodes. The structure learning algorithms used by BNW guarantee the discovery of the best (most probable) network structure given the data. To facilitate network modeling across multiple biological levels, BNW provides a very flexible interface that allows users to assign network nodes into different tiers and define the relationships between and within the tiers. This function is particularly useful for modeling systems genetics datasets that often consist of multiscalar heterogeneous genotype-to-phenotype data. BNW enables users to, within seconds or minutes, go from having a simply formatted input file containing a dataset to using a network model to make predictions about the interactions between variables and the potential effects of experimental interventions. In this chapter, we will introduce the functions of BNW and show how to model systems genetics datasets with BNW.


Round ligament varicosities associated with uterine varicosities in pregnancy: a case report.
Round ligament varicosities associated with uterine varicosities are rare during pregnancy. We report the case of a pregnant woman with a painful mass in her left groin at 32 weeks' gestation. Ultrasound revealed multiple distended veins within the left round ligament and over the uterus. The patient was treated by close observation and had a vaginal delivery of a healthy baby.


[Endosseous-endodontic implants: indications and technic].
The endodontic endosseous implants were used to a lot of clinic indication. The Authors suggest their use in the median fractures of roots and describe the operative technic, of which they consider the long-term results.


Direct NMR-spectroscopic determination of active-enzyme concentration by titration with a labeled inhibitor: determination of the k(cat) value of almond beta-glucosidase.
A new method for the determination of active-enzyme concentration of a glucosidase by using (13)C NMR spectroscopy is reported. The method consists of quantifying the binding between a (13)C-labelled, strong competitive inhibitor, [5-(13)C]-1-azafagomine (1), and the enzyme. The concentration of free inhibitor 1 is measured in a series of binding experiments from the intensity of its NMR signal relative to that of a reference. From a plot of the concentrations of bound vs. free inhibitor 1, the amount of specifically bound 1, that is, the amount of active sites, is determined. From this value, active-enzyme concentration and k(cat) value can be calculated.


De novo FUS mutations are the most frequent genetic cause in early-onset German ALS patients.
In amyotrophic lateral sclerosis (ALS) patients with known genetic cause, mutations in chromosome 9 open reading frame 72 (C9orf72) and superoxide dismutase 1 (SOD1) account for most familial and late-onset sporadic cases, whereas mutations in fused in sarcoma (FUS) can be identified in just around 5% of familial and 1% of overall sporadic cases. There are only few reports on de novo FUS mutations in juvenile ALS patients. To date, no systematic evaluation on the frequency of de novo FUS mutations in early-onset ALS patients has been conducted. Here, we screened a cohort of 14 early-onset sporadic ALS patients (onset age <35 years) to determine the frequency of mutations in C9orf72, SOD1, and FUS in this defined patient cohort. All patients were recruited prospectively by a single center in a period of 38 months. No mutations were detected in SOD1 or C9orf72; however, we identified 6 individuals (43%) carrying a heterozygous FUS mutation including 1 mutation that has not been described earlier (c.1504delG [p.Asp502Thrfs*27]). Genetic testing of parents was possible in 5 families and revealed that the mutations in these patients arose de novo. Three of the 6 identified patients presented with initial bulbar symptoms. Our study identifies FUS mutations as the most frequent genetic cause in early-onset ALS. Genetic testing of FUS thus seems indicated in sporadic early-onset ALS patients especially if showing predominant bulbar symptoms and an aggressive disease course.


A Fano-resonance plasmonic assembly for broadband-enhanced coherent anti-Stokes Raman scattering.
Surface-enhanced coherent anti-Stokes Raman scattering (SECARS) technique has triggered huge interests due to the significant signal enhancement for high-sensitivity detection. Previous SECARS work has tended to focus only on the enhancement effect at a certain combination of frequencies, more suitable for single-frequency CARS. In this work, based on the enhancement factor for broadband SECARS excitation process, a novel Fano resonance plasmonic nanostructure for SECARS is studied. In addition to the 12 orders of magnitude enhancement effect that can be realized under single-frequency CARS, this structure also shows huge enhancement under broadband CARS in a wide wavenumber region, covering most of the fingerprint region. This geometrically-tunable Fano plasmonic nanostructure provides a way to realize broadband-enhanced CARS, with potentials in single-molecular monitoring and high-selectivity biochemical detection.


An Overview of Parotidectomy for Benign Parotid Lesions with Special Reference to Perioperative Techniques to Avoid Complications: Our Experience.
The aim of this analytical study was to assess the reduction in intra- and post-operative complications of parotid surgeries for benign lesions following simple surgical steps and to compare the outcome with the literature. A retrospective study of 77 patients requiring parotidectomy surgery for benign parotid lesions was conducted in Department of Otorhinolaryngology, Sri Guru Ram Rai Institute of Medical and Health Sciences, Dehradun from the period of January 2011 to January 2016. Patients were followed up for 1-5 years and serial evaluation was done for all cases who developed any of intra- or post-operative complications in their disease course. The ages of presentation varied from 1.5 to 70 years. There were 51 males and 26 female patients. Majority cases were of pleomorphic adenoma followed by warthins tumor. Superficial parotidectomy was done in 69 patients; 4 cases required near total parotidectomy while total conservative parotidectomy was done in 4 cases. The commonest complication was altered sensations around the ear which was seen in 10 cases followed by temporary facial nerve paresis and sialocele formation which occurred in 3 cases each. This study advocates the importance of detailed anatomical knowledge, behavior of various benign pathologies, experience of surgeon and his fine dissection techniques to ensure minimum risks of injury to important structures in surgical field.


Radiation therapy improves CAR T cell activity in acute lymphoblastic leukemia.
Autologous T cells engineered to express a chimeric antigen receptor (CAR) specific for CD19 are approved for the treatment of various CD19+ hematological malignancies. While CAR T cells induce objective responses in a majority of patients, relapse frequently occurs upon loss of CD19 expression by neoplastic cells. Radiation therapy (RT) has been successfully employed to circumvent the loss of CAR targets in preclinical models of pancreatic cancer. At least in part, this reflects the ability of RT to elicit death receptor (DR) expression by malignant cells, enabling at least some degree of CAR-independent tumor killing. In a human model of CD19+ acute lymphoblastic leukemia (ALL), we also observed DR upregulation by RT, both in vitro and in vivo. Moreover, low-dose total body irradiation (LD-TBI) delivered to ALL-bearing mice prior to CAR T cell infusion considerably extended the overall survival benefit afforded by CAR T cells alone. Such an improved therapeutic activity was accompanied by a superior expansion of CAR T cells in vivo. These data encourage the initiation of clinical trials combining LD-TBI with CAR T cells in patients with hematological malignancies.


One number does not fit all: mapping local variations in resolution in cryo-EM reconstructions.
The resolution of density maps from single particle analysis is usually measured in terms of the highest spatial frequency to which consistent information has been obtained. This calculation represents an average over the entire reconstructed volume. In practice, however, substantial local variations in resolution may occur, either from intrinsic properties of the specimen or for technical reasons such as a non-isotropic distribution of viewing orientations. To address this issue, we propose the use of a space-frequency representation, the short-space Fourier transform, to assess the quality of a density map, voxel-by-voxel, i.e. by local resolution mapping. In this approach, the experimental volume is divided into small subvolumes and the resolution determined for each of them. It is illustrated in applications both to model data and to experimental density maps. Regions with lower-than-average resolution may be mobile components or ones with incomplete occupancy or result from multiple conformational states. To improve the interpretability of reconstructions, we propose an adaptive filtering approach that reconciles the resolution to which individual features are calculated with the results of the local resolution map.


Trichodermatides A-D, four new polyketides from Trichoderma sp. XM-3.
Four new polyketides named trichodermatides A-D (1-4), along with five known analogues (5-9), were isolated from the fungus Trichoderma sp. XM-3. Their structures were elucidated on the basis of HRESIMS and NMR analyses, and their absolute configurations were determined by ECD comparison, 1H and 13C NMR calculation, DP4+ analysis, modified Mosher's method, and X-ray crystallography. Trichodermaketone D (9) showed mild antibacterial activity against Pseudomonas aeruginosa.


A first calibration of culturable bacterial diversity and their dual resistance to heavy metals and antibiotics along altitudinal zonation of the Teesta River.
Culturable bacterial diversity and co-occurrence of heavy metal and antibiotic resistance were investigated from the water and sediments along the course of the Teesta River, in the Eastern Himalayas. Water and sediment samples collected from six sampling points during the monsoon and winter seasons were subjected to analysis of physico-chemical parameters, heavy metal contamination and antibiotic tolerance. The culturable bacterial diversity established by application of bacterial culture and 16S rRNA gene sequencing, ascertained the majority belonged to Proteobacteria, Actinobacteria, Firmicutes, Bacteroidetes and Deinococcus-Thermus. Among the 5 phyla, Proteobacteria and Actinobacteria were the dominant phyla present in both water and sediment samples, whereas Bacteroidetes, Firmicutes and Deinococcus-Thermus were unique to particular sites. The Shannon index indicated that the bacterial richness was more in the water column as compared to sediment. From the total of 245 isolates, 69 genera were identified. Heavy metal tolerance and antibiotic resistance profiles showed some isolates to be tolerant to high levels of heavy metals and multiple antibiotics indicating a major concern in terms of river ecosystem serving as a pool for dissemination of such resistant genes. The antibiotic resistance and heavy metal contamination diversified along the human-impacted downstream sites, endorsing the contribution of anthropogenic factors. The present report on bacterial diversity and the associated metal and antibiotics tolerance among bacteria is the first of its kind on Teesta River, the only major river system flowing through the state of Sikkim and parts of North Bengal.


The nature of the initial step in the conformational folding of disulphide-intact ribonuclease A.
Here we investigate conformational folding reaction of disulphide-intact ribonuclease A in the absence of the complicating effects due to non-native interactions (such as cis/trans proline isomerization) in the unfolded state. The conformational folding process is found to be intrinsically very fast occurring on the milliseconds time scale. The kinetic data indicate that the conformational folding of ribonuclease A proceeds through the formation of a hydrophobically collapsed intermediate with properties similar to those of equilibrium molten-globules. Furthermore, the data suggest that the rate-limiting transition states on the unfolding and refolding pathways are substantially different with the refolding transition state having non-native-like properties.


Development of a rapid total nucleic acid extraction method for the isolation of hepatitis A virus from fresh produce.
Recently, there have been increasing reports of foodborne illnesses associated with the consumption of fresh produce. Among these, hepatitis A virus (HAV) remains epidemiologically important and has been continually implicated in several outbreaks. We describe a rapid method (<8h) for the isolation and subsequent detection with real-time quantitative PCR (RT-qPCR) of the HAV HM-175 cytopathic strain seeded onto baby spinach and sliced tomatoes using a total RNA extraction method, utilizing a high concentration (4M) guanidine thiocyanate buffer. Consistent detection of HAV genome from both produce items was achieved at an inoculation level of 3×10³ PFU/25 g of food, with less consistent detection achieved at 3×10² PFU/25g. Initial studies revealed that a final precipitation of recovered RNA with potassium acetate to reduce carryover of polysaccharides and the addition of polyvinylpyrrolidone to remove polyphenolics in spinach were essential. For tomatoes, virus isolation was achieved with the incorporation of either an elution step with a high pH Tris-glycine-beef extract (TGBE) buffer or with an enzymatic digestion with pectinase. We also describe the development of a protocol for the detection of HAV from tomatoes utilizing a Luminex® microbead-based suspension array. The results correlated well with the RT-qPCR assay suggesting the feasibility of the Bioplex® as a detection platform for viruses isolated from foods.


Prevalence and influencing factors of chronic pain in middle-aged and older adults in China: results of a nationally representative survey.
With China's rapidly aging population, chronic pain has become a major public health issue. This article aims at determining associations between chronic pain and multiple factors, including demographic characteristics, health status, and health service utilization of middle-aged and older adults in China. We selected all the 19,829 respondents who were over 45 years old from the China Health and Aging Tracking Survey 2018 (CHARLS) as our study population. The key information in terms of the body pain, demographic characteristics, health status, behaviors and health services use was extracted and analyzed. Logistic regression model was used to determine the influencing factors of chronic pain. Analysis revealed that 60.02% (9,257) of the data from this survey reported physical pain, with pain sites concentrated at the head (40.9%), lower back (62.2%) and knees (47.2%). Pain was positively associated with influencing factors for pain: being a female (OR = 2.10, 95% CI 1.90-2.33, p < 0.001), living in a western region (OR = 1.28, 95% CI 1.16-1.41, p < 0.001), living in a rural area (OR = 1.14, 95% CI 1.06-1.23, p < 0.001), smoked (OR = 1.26, 95% CI 1.14-1.38, p < 0.001), drank alcohol (OR = 1.16, 95% CI 1.06-1.26, p = 0.001), and had poor self-rated health (OR = 6.84, 95% CI 5.41-8.65, p < 0.001), had hearing problems (OR = 1.23, 95% CI 1.11-3.37, p < 0.001), were depressed (OR = 1.56, 95% CI 1.03-1.29, p < 0.001), had arthritis (OR = 2.21, 95% CI 2.02-2.41, p < 0.001), stomach disorders (OR = 1.69, 95% CI 1.55-1.85, p < 0.001), visited a Western medicine hospital (OR = 1.28, 95% CI 1.10-1.50, p = 0.002), and visits to other medical institutions (OR = 1.42, 95%CI 1.22-1.64, p < 0.001). On the other side, as a protective factor for pain, having nighttime sleep ≥7 h (OR = 0.74, 95%CI 0.68-0.80, p < 0.001) was negatively associated with pain. Physical pain affects many older adults. Women, regional, rural residents, smokers, alcohol drinkers, people with poor self-rated health, those having <7 h of sleep at night, those with hearing problems, depression, arthritis, stomach disorders, and people who visits Western hospitals or other medical institutions are at greater risk for pain and deserve the attention of health care providers and policy makers to focus on pain prevention and management in middle-aged and older adults. Future research studies should also focus on the impact of health literacy on pain prevention and management outcomes.


HIF-1α drives resistance to ferroptosis in solid tumors by promoting lactate production and activating SLC1A1.
Solid tumors have developed robust ferroptosis resistance. The mechanism underlying ferroptosis resistance regulation in solid tumors, however, remains elusive. Here, we report that the hypoxic tumor microenvironment potently promotes ferroptosis resistance in solid tumors in a hypoxia-inducible factor 1α (HIF-1α)-dependent manner. In combination with HIF-2α, which promotes tumor ferroptosis under hypoxia, HIF-1α is the main driver of hypoxia-induced ferroptosis resistance. Mechanistically, HIF-1α-induced lactate contributes to ferroptosis resistance in a pH-dependent manner that is parallel to the classical SLC7A11 and FSP1 systems. In addition, HIF-1α also enhances transcription of SLC1A1, an important glutamate transporter, and promotes cystine uptake to promote ferroptosis resistance. In support of the role of hypoxia in ferroptosis resistance, silencing HIF-1α sensitizes mouse solid tumors to ferroptosis inducers. In conclusion, our results reveal a mechanism by which hypoxia drives ferroptosis resistance and identify the combination of hypoxia alleviation and ferroptosis induction as a promising therapeutic strategy for solid tumors.


Core muscle functional strength training for reducing the risk of low back pain in military recruits: An open-label randomized controlled trial.
Core muscle functional strength training (CMFST) has been reported to reduce injuries to the lower extremity. However, no study has confirmed whether CMFST can reduce the risk of low back pain (LBP). This study identified the effects of CMFST on the incidence of LBP in military recruits. We performed a prospective, open-label, randomized, controlled study in a population of young healthy male naval recruits from a Chinese basic combat training program. Participants were randomly assigned to either the core group or the control group. In additional to normal basic combat training, recruits in the core group underwent a CMFST program for 12 weeks, while recruits in the control group received no extra training. At the beginning of the study and at the 12th week, the number of participants with LBP was counted, and lumbar muscle endurance was measured. In addition, when participants complained of LBP, they were assessed using the visual analog scale (VAS) and Roland Morris Disability Questionnaire (RMDQ). A total of 588 participants were included in the final analysis (295 in the core group and 293 in the control group). The incidence of LBP in the control group was about twice that of the core group over the 12-week study (20.8% vs 10.8%, odds ratio: 2.161-2.159, P < 0.001). The core group had better lumbar muscle endurance at 12 weeks than the control group ([200.80 ± 92.98] s vs [147.00 ± 84.51] s, P < 0.01). There was no significant difference in VAS score between groups, but the core group had a significantly lower RMDQ score at week 12 than the control group (3.33 ± 0.58 vs 5.47 ± 4.41, P < 0.05). This study demonstrated that the CMFST effectively reduced the incidence of LBP, improved lumbar muscle endurance, and relieved the dysfunction of LBP during basic military training.


A new species of Platydecticus (Orthoptera: Tettigoniidae: Tettigoniinae; Nedubini) from the Andes of Chile.
A new species of the genus Platydecticus is described based on adult male and female specimens and the egg. The new species, Platydecticus diaguita, inhabits the Andes Range at 27º S latitude, above 3000 m elevation. Both sexes are easily identifiable by genital morphology characters and by the external characters of the fastigium of the vertex and the reduced number of spines in the hind tibia. It is also the smallest species described for the genus.


Ureteric obstruction secondary to endometriosis. Report of three cases with a review of the literature.
Three cases of endometriosis are presented. In one patient bilateral obstruction was noted; in a second patient bilaterally enlarged kidneys with unilateral obstruction were found and in the third patient only unilateral obstruction was defined. Intrinsic ureteric obstruction and bilateral obstruction, both rare findings, are described in this report. The diagnosis, evaluation and management of these patients are discussed.


Motion-corrected 23 Na MRI of the human brain using interleaved 1 H 3D navigator images.
To evaluate the feasibility of motion correction for sodium (23 Na) MRI based on interleaved acquired 3D proton (1 H) navigator images. A 3D radial density-adapted sequence for interleaved 23 Na/1 H MRI was implemented on a 7 Tesla whole-body MRI system. The 1 H data obtained during the 23 Na acquisition were used to reconstruct 140 navigator image volumes with a nominal spatial resolution of (2.5 mm)3 and a temporal resolution of 6 s. The motion information received from co-registration was then used to correct the 23 Na image dataset, which also had a nominal spatial resolution of (2.5 mm)3 . The approach was evaluated on six healthy volunteers, whose motion during the scans had different intensities and characteristics. Interleaved acquisition of two nuclei did not show any relevant influence on image quality (SNR of 13.0 for interleaved versus 13.2 for standard 23 Na MRI and 176.4 for interleaved versus 178.0 for standard 1 H MRI). The applied motion correction increased the consistency between two consecutive scans for all examined volunteers and improved the image quality for all kinds of motion. The SD of the differences ranged between 2.30% and 6.96% for the uncorrected and between 2.13% and 2.67% for the corrected images. The feasibility of interleaved acquired 1 H navigator images to be used for retrospective motion correction of 23 Na images was successfully demonstrated. The approach neither affected the 23 Na image quality nor elongated the scan time and can therefore be an important tool to improve the accuracy of quantitative 23 Na MRI.


COVID-19 and attitudes towards early withdrawal of pension funds: The role of trust and political ideology.
In response to the COVID-19 pandemic, the Chilean Congress approved three laws between July 2020 and April 2021 that allowed early withdrawals of pension funds without any eligibility constraints. In this paper, we use nationwide survey data to examine the factors associated with people's assessments about the suitability of these policies in the context of the pandemic, with a particular focus on trust and political ideology. We find that respondents that declare high levels of trust in Chile's pension system, financial system, and political institutions and actors tend to oppose early withdrawal policies. Similarly, respondents on the right of the political spectrum and those that voted for the incumbent president, who opposed this policy, also declare opposition to early withdrawals. Overall, our findings suggest that political attitudes and beliefs are associated with policy views on changes to Chile's pension-fund system and that support for early withdrawal policies may be driven by lack of confidence in institutions.


Comparison of stereofundus photographs in patients with insulin-dependent diabetes during conventional insulin treatment or continuous subcutaneous insulin infusion.
Sixty-five patients with mild to moderate nonproliferative diabetic retinopathy who enrolled in a prospective controlled clinical trial had stereofundus photographs assessed for change over an 8-mo period. The entire study group showed a worsening of retinopathy with time (P less than 0.001). The worsening was greater in the pump-treated group (15/32) than in the conventionally treated group (9/33). The significance of this difference ranged from P = 0.67, if changes in mean retinopathy level for each patient were compared, to P = 0.177 if a grading system keyed to the worse eye was compared. The difference in rates of change between treatment groups was found to be related to the baseline mean retinopathy level (P = 0.031), but less significantly so if baseline retinopathy keyed to the worse eye was used as a covariate (P = 0.08). Worsening occurred more frequently in those patients starting with the lower retinopathy levels. Progression was associated with the appearance of retinal infarcts (cotton-wool spots, soft exudates) and/or intraretinal microvascular abnormalities, with the pump patients showing a significant increase in these individual retinal lesions compared with the conventionally treated patients over 8 mo (P less than 0.025).


Pharmacoepidemiology for oncology clinical practice: Foundations, state of the art and perspectives.
Since the early 2000s, the arrival of the so-called targeted therapies and immunotherapies have prolonged survival rates in many cancers. In parallel, post-marketing surveillance of anticancer drugs through pharmacoepidemiology has gradually developed. This paper provides (i) a detailed argumentation of the foundations for pharmacoepidemiology of anticancer drugs, (ii) an overview of pharmacoepidemiological studies currently available in this field, and (iii) some perspectives to improve pharmacoepidemiology for oncology practice. First of all, according to the existing literature, the development of pharmacoepidemiological studies for the clinical evaluation of anticancer drugs appears particularly justified based on common limitations of clinical trials in oncology regarding essential methodological principles such as adequate control groups, randomisation or double blinding. Many descriptive field cohort studies have investigated together treatment patterns, effectiveness, and safety to compare results from clinical trials with those of everyday practice. The utilisation of anticancer drugs has also been extensively described through cross-sectional or cohort studies by often using medico-administrative or medical databases. Such studies are useful to quantify and characterise use over time in the population, including clinically unvalidated use, and to evaluate adherence and persistence to increasingly available oral anticancer drugs. Despite their importance to increase knowledge, comparative effectiveness or safety studies remain uncommon. In a context of rapidly emerging therapies and personalised treatments, this may be due to methodological challenges especially related to the choice of a comparator or the consideration of confounding by indication. In the future, efforts must be pursued to provide real-time access to high-quality, large-scale clinical, biological and treatment data, and to improve record-linkage between hospital and outpatient databases. More research is also needed to better evaluate all medications, not only anticancer, as part of an overall cancer care pathway and to bring the evaluation of anticancer drugs closer to patients and society (social pharmacology).


[Reconstruction of the middle phalangeal base of the finger using an osteocartilaginous autograft from the hamate].
Restoration of proximal interphalangeal joint stability with preservation of mobility by reconstruction of the middle phalanx base using an osteochondral graft from the carpometacarpal joint surface of the hamate. Acute and older isolated destruction of the palmar middle phalanx base >25%. Destruction of the head of the proximal phalanx, advanced chondropathia of the head of the proximal phalanx, extensive soft tissue injury with loss of skin coverage at the proximal interphalangeal joint. The fractured middle phalangeal base is debrided and the defect is replaced by a size-matched autograft from the dorsal carpometacarpal hamate osteoarticular surface that is secured in place with miniscrews. Immobilization for 2 weeks in a below-elbow cast in intrinsic plus position. Subsequent immobilization by a splint including the distal and proximal interphalangeal joint. Healing was achieved in 100% with restoration of joint congruity in 12 of 13 cases and slight subluxation in 1 case. Follow up was possible in 9 cases after 22 ± 16 (5-51) months. The average range of motion in the reconstructed joint achieved 0/9/73°, grip strength 82% of the unaffected side. Of the 9 patients, 5 developed a mild flexion contracture of the proximal interphalangeal joint. The DASH score achieved 4 ± 3 (0-8) points, pain at rest was 1 ± 2 (0-5), pain at exercise 2 ± 2 (0-5) on a visual analogue scale (0-10). All patients were satisfied and willing to undergo the procedure again. According to the literature, reconstruction of the base of the middle phalanx by using an osteochondral graft from the hamate is a reliable procedure to restore stability and mobility of the joint.


A left hemisphere contribution to visuospatial processing.
Men with chronic, focal, unilateral missile injures of the brain--twenty-five with left hemisphere lesions (LH group) and twenty with right hemisphere lesions (RH groups)--and twenty-two control subjects were given two visuoperceptual and two visuospatial tests. The LH group was significantly impaired in relation to the control group on both the spatial tasks. A different pattern of dissociable perceptual and spatial deficits was found in the experimental groups: better preserved perceptual than spatial performance was observed more frequently in the LH group whereas the converse--relatively better spatial than perceptual performance--was more evident in the RH group. Double dissociations in performance on the two spatial tasks were found predominantly in the LH group. These findings suggest an important left hemisphere contribution to visuospatial processing and the possibility of a more focal representation of spatial abilities in the left hemisphere than in the right.


Transient hyper-17-OHPnemia unrelated to cross-reactions with residual fetal adrenal cortex products.
To clarify the pathogenesis of transient hyper-17alpha-hydroxyprogesteronemia, we initiated a laboratory investigation in a pre-term infant with persistently high serum 17alpha-hydroxyprogesterone (17-OHP) until 2 months of age. Serum 17-OHP level was measured by high-performance liquid chromatography and radioimmunoassay, and gene analysis of CYP21A2 (21-hydroxylase) was performed. Serum 17-OHP level on the 29th day of life was 25.4 ng/ml, and the urinary steroid profile showed low pregnanetriolone. Gene analysis of 21-hydroxylase disclosed no mutation, and 17-OHP normalized by 3 months of age without specific treatment. Transient elevations in 17-OHP, which do not appear related to cross-reactions with products of a residual fetal adrenal cortex, may occur in the first few months of life.


Trial of a new acellular pertussis vaccine in healthy adult volunteers.
Immunogenicity and reactogenicity of a new acellular pertussis vaccine were tested in healthy adults. The vaccine contained three constituents of Bordetella pertussis; filamentous haemagglutinin, pertussis toxin (PT) and fimbriae bearing agglutinogens 2 and 3. The constituents were separately purified, treated with formaldehyde and combined with one of two aluminium adjuvants. Subjects received one dose of vaccine or an appropriate adjuvant-only preparation and were monitored for clinical responses for 7 days. Results with the two forms of vaccine were similar. Of 35 vaccinees, none had a temperature higher than 37 degrees C or a severe reaction, one had a moderate reaction (possibly due in part to intercurrent infection) and nine had mild reactions confined to localized discomfort and/or erythema or induration at the injection site. All vaccinees had good serum antibody responses to vaccine antigens measured by ELISA and for PT, by neutralization of its effects on Chinese hamster ovary cells.


Impact of dexamethasone-induced immunosuppression on the duration and level of shedding of Escherichia coli O157:H7 in calves.
The goal of this study was to determine whether immunosuppression plays a role in the level and duration of fecal shedding of Escherichia coli O157. Immunosuppression was induced in calves by administering dexamethasone. Six 1-week-old Holstein bull calves were injected intramuscularly with dexamethasone and orally inoculated with 10(9) CFU of a mixture of three nalidixic-acid resistant strains of E. coli O157:H7. Five 1-week-old Holstein bull calves that were given the same oral inoculation of E. coli O157:H7, but not the dexamethasone injections, served as controls. All calves were examined daily and fecal samples were collected three times a week for detection and enumeration of the nalidixic-acid resistant E. coli O157. Four weeks after the last calf stopped shedding, all calves were necropsied and samples from the gastrointestinal tract were taken for the detection of the nalidixic-acid resistant E. coli O157. Dexamethasone-injected calves shed at higher levels (P = 0.04) on days 4 and 7 postinoculation, but not thereafter. None of the samples collected at necropsy were positive for E. coli O157. Data from this study suggest that there may be a time-dependent relationship between dexamethasone immunosuppression and the fecal concentration of E. coli O157 but that transient immunosuppression does not appear to prolong shedding of E. coli O157.


Effect of helium/neon laser irradiation on nerve growth factor synthesis and secretion in skeletal muscle cultures.
Low energy laser irradiation therapy in medicine is widespread but the mechanisms are not fully understood. The aim of the present study was to elucidate the mechanism by which the light might induce therapeutic effects. Skeletal muscle cultures were chosen as a target for light irradiation and nerve growth factor (NGF) was the biochemical marker for analysis. It was found that there is a transient elevation of intracellular calcium in the myotubes immediately after irradiation (P<0.001). Preincubation of the myotubes with either the photosensitizers 5-amino-levulinic acid (5-ALA), or with hematoporphyrin (Hp) enhanced the elevation of cytosolic calcium (P<0.001) after helium/neon irradiation (633 nm) with an energy of 3 J/cm(2). In addition, helium/neon irradiation augmented the level of NGF mRNA fivefold and increased NGF release to the medium of the myotubes. Thus, it is speculated that transient changes in calcium caused by light can modulate NGF release from the myotubes and also affect the nerves innervating the muscle. The NGF is probably responsible for the beneficial effects of low-level light.


First Case of Human Brucella canis Infection in the Netherlands.
A patient was diagnosed with Brucella canis following exposure to infected dogs in her breeding facility. Transboundary spread of B. canis through (illegal) import of infected dogs to non-endemic countries in Europe suggest that B. canis infection should be considered in European patients with occupational exposure to dogs.


Retraction to artesunate induces apoptosis and inhibits the proliferation, stemness, and tumorigenesis of leukemia.
[This retracts the article DOI: 10.21037/atm-20-4558.].


The impact of adjacent segment degeneration on the clinical outcome after lumbar spinal fusion.
A retrospective study. To determine the impact of the adjacent segment degeneration (ASD) on the clinical outcome after lumbar fusion. Few studies have analyzed correlation between clinical outcomes and ASD. From January 2000 to December 2004, 217 patients who underwent lumbar fusion and have more than 2 years of follow-up were evaluated. The patients were divided into 3 groups: fusion was carried out on 1 (A), 2 (B), and more than 3 segments (C). UCLA grading scale was used to evaluate the ASD on radiograph preoperative, postoperative, and last follow-up. The clinical outcome was evaluated and the association between clinical outcome and ASD was evaluated by the Spearman correlation. ASD occurred 11.6% (13/112 cases) in group A, 14.5% (9/62 cases) in group B, and 16.3% (7/43 cases) in group C. Among them, the cases which had progressed more than 2 level of degenerative grade were 0 in group A, 7 cases (77.8%) in group B, and 6 cases (85.7%) in group C. In group A, 5 cases with ASD had down graded 1 level in clinical outcome. In group B, all 9 cases with ASD had down graded more than 1 level in clinical outcome; 1 level aggravation in 1, 2 level in 5, and 3 level in 1 case. In group C, all 7 cases with ASD also had down graded more than 1 level in clinical outcome; 1 level aggravation in 2, 2 level in 5 cases. Finally, the more the change of the degenerative grade is severe, the more the clinical outcome is worse (P < 0.05). The impact of ASD on clinical outcome after fusion showed a significant correlation, especially with the ASD after multiple-segments fusion. Careful attention is needed in multiple-segments fusion such as degenerative lumbar deformity.


Protein synthesis and fast axonal transport in regenerating goldfish retinal ganglion cells.
To characterize the fast component of axonal transport in regenerating goldfish optic axons, the incorporation of L-2,3-[3H]proline into newly-synthesized proteins in the cell bodies of the retinal ganglion cells and the amount of transported labeled protein were determined at 2-36 days after cutting the optic tract. Both the incorporation and the amount of transported protein had doubled by 10 days after the lesion and continued to increase to about 5 times normal at 15 days, a time when a large proportion of the regenerating axon population had reached the optic tectum. Near-normal levels were recovered by 36 days. In contralateral control neurons, the incorporation of L-2,3-[3H]proline was unchanged from normal throughout, whereas the amount of labeled transported protein entering control axons was decreased by 55% at 2 and 10 days after the testing lesion, returning to normal by 15 days. An increase in fast transport velocity was seen in the regenerating axons beginning at 10 days after the lesion. However, a similar velocity increase was also seen in the contralateral control axons and in undamaged axons following removal of the cerebral hemispheres. Therefore, the velocity increase was not a specific consequence of axotomy.


The aging of the endocrine pancreas of the rat. I. Parameters of cell proliferation.
Morphometrical analysis of the endocrine pancreas of senile 30-month-old rats revealed that the volume density as well as the numeric density of islets of Langerhans were much lower than in 24-month-old rats, which coincided with a much higher percentage of pycnotic nuclei in islet cells. The proportion and localization of the different categories of endocrine cells (A, B, D and PP) remained however unchanged with aging. The apparent problem of cell renewal observed in vivo in the very old age was detected earlier in vitro by tritiated thymidine incorporation. Such experiments showed that 24-month-old islet cells had a decreased labelling index when compared to 3-month-old cells. The proliferation capacity of the old cells could be partially increased by changing the serum concentration or type. Similarly as being more sensitive to serum factors, these cells underwent also more pronounced negative influence of high oxygen pressure on replication. A stereological analysis of the ultrastructure of non-degenerated B-cell nuclei revealed that with age, the relative volume of the condensed chromatin increased progressively at the expense of the dispersed form. This suggests that the still functioning senile B-cells could reduce their transcriptional activity.


Photosynthetic reaction centre of Chloroflexus aurantiacus. Primary structure of M-subunit.
The M-subunit primary structure of the reaction centre (RC) from Chloroflexus aurantiacus composed of 306 amino acid residues has been determined by parallel analysis of the protein and corresponding DNA. The blocked N-terminus as well as replacement of the essential histidine liganding Mg of an accessory bacteriochlorophyll in purple bacteria by leucine distinguishes the M-subunit of Chloroflexus RC from that of purple bacteria.


Living with an inborn error of metabolism detected by newborn screening-parents' perspectives on child development and impact on family life.
Newborn screening for inborn errors of metabolism is regarded as highly successful by health professionals. Little is known about parents' perspectives on child development and social impact on families. Parents of 187 patients with metabolic disorders detected by newborn screening rated child development, perceived burdens on child and family, and future expectations on a questionnaire with standardized answers. Parental ratings were compared with standardized psychometric test results. Regression analysis was performed to identify factors associated with extent of perceived burden. In 26.2% of patients, parents perceived delays in global development and/or specific developmental domains (physical, social, intellectual, language). Parents expected normal future development in 95.7%, and an independent adult life for their child in 94.6%. Comparison with psychometric test results showed that parents of children with cognitive impairments tended to overrate their child's abilities. Mild/medium burden posed on the family (child) by the metabolic disorder was stated by 56.1% (48.9%) of parents, severe/very severe burden by 19.3% (8.6%). One third of families reported financial burden due to the metabolic disorder. Dietary treatment and diagnoses with risk for metabolic decompensation despite treatment were associated with higher perceived burden for the family. Disorders rated as potentially very burdensome by experts were not rated accordingly by parents, demonstrating different perspectives of professionals and parents. Although newborn screening leads to favourable physical and cognitive outcome, living with a metabolic disorder may cause considerable stress on patients and families, emphasizing the need for comprehensive multidisciplinary care including psychological and social support.


Shape-dependent enzyme-like activity of Co3O4 nanoparticles and their conjugation with his-tagged EGFR single-domain antibody.
In this study, Co3O4 nanopolyhedrons, nanocubes, nanoplates and nanorods were synthesized and characterized. Furthermore, the peroxidase- and catalase-like activities of these Co3O4 nanoparticles (NPs) were studied and influence of the exposed crystal planes was explored. According to their morphology and peroxidase-like activity, dimercaptosuccinic acid (DMSA) modified Co3O4 nanopolyhedrons synthesized via coprecipitation method (Co3O4 NHs) were selected as a proper candidate for the immunohistochemical (IHC) detection of epidermal growth factor receptor (EGFR) expression in non-small cell lung cancer (NSCLC) tissues. Bivalent cobalt ions were coupled to the carboxyls on the surface of the obtained Co3O4 NHs so as to chelate the hexahistidine residues (His-Tags) at the C-terminal of EGFR single-domain antibodies (EGFR sdAbs). Finally, the as-obtained EGFR sdAbs-binding Co3O4 NHs (Co3O4 nanoprobes) were successfully applied to the detection of EGFR expression in NSCLC tissues.


Effect of tissue-plasminogen activator on leukocyte-endothelial interactions at the microcirculatory level.
In free tissue transfer and replantation surgery, there is a debate over whether any pharmacologic agents should be used to improve vessel patency and tissue survival. Because tissue-plasminogen activator (t-PA) is a highly effective and safe fibrinolytic, it may be useful in obtaining and maintaining vessel patency. The direct effects of t-PA on skeletal muscle hemodynamics and leukocyte activation at the microcirculatory level were investigated. Male Sprague-Dawley rats (n = 20) were divided into three experimental groups: control (n = 8), vehicle (n = 6), and t-PA (n = 6). Using the cremaster muscle flap model and intravital microscopy, red blood cell velocity, vessel diameter, capillary perfusion, endothelial edema index, and leukocyte-endothelial interactions (rolling, adhering, and transmigrating leukocytes) in postcapillary venules were measured. In the vehicle and t-PA groups, vehicle or t-PA was infused by means of a catheter inserted into the lower abdominal aorta for local infusion. Except for a significant reduction in the diameter of the first order arterioles from 117 microm to 82 microm (medians; p = 0.026), t-PA did not significantly affect red blood cell velocity, vessel diameter, or capillary perfusion compared with vehicle. However, leukocyte-endothelial interactions did differ significantly in postcapillary venules. Adhering leukocytes counted per visual field decreased from 4.67 in the vehicle group and 3.50 in the control group to 1.67 in the t-PA group (medians; p = 0.015 and p = 0.005, respectively); transmigrating leukocytes in the t-PA group decreased from 4.75 in the vehicle group and 3.50 in the control group to 1.67 in the t-PA group (medians; p = 0.002 and p = 0.043, respectively). t-PA treatment significantly decreased the number of both adhering and transmigrating leukocytes. These novel findings on leukocyte-endothelial interactions suggest that t-PA has anti-inflammatory effect.


Familial acanthosis nigricans with madarosis.
We report familial acanthosis nigricans affecting a 35-year-old woman, her 7-year-old son and 5-year-old daughter. Absence of the eyebrows and eyelashes was noted in this family. The mother had no axillary hair and her pubic hair was sparse. The boy also suffered from congenital heart disease and a congenital cataract in the left eye. The combination of acanthosis nigricans and ectodermal defects in this family may represent a distinct nosological entity.


Protein function microarrays for customised systems-oriented proteome analysis.
Protein microarrays have many potential applications in the systematic, quantitative analysis of protein function. However, simple, reproducible, and robust methods for array fabrication that are compatible with the study of large, custom collections of potentially unrelated proteins are required. Here, we discuss different routes to array fabrication and describe in detail one approach in which the purification and immobilisation procedures are combined into a single step, significantly simplifying the array fabrication process. We illustrate this approach by reference to the creation of an array of human protein kinases and discuss methods for assay and data analysis on such arrays.


The CD11/CD18 leukocyte glycoprotein deficiency.
CD11/CD18 leukocyte glycoprotein deficiency is a rare, inherited disorder of leukocyte function, manifested by recurrent severe bacterial infections. A deficiency in the expression of a family of leukocyte membrane glycoproteins (the CD11/CD18 glycoproteins) represents the molecular basis for this disease.


Temporal heterogeneity of placental segmental fetal vascular malperfusion: timing but not etiopathogenesis.
Clinicopathologic correlations of segmental villous avascularity and other histological lesions of segmental fetal vascular malperfusion (SFVM) were analyzed retrospectively to determine whether lesions of various durations reflect different etiopathogeneses. The frequencies of 25 independent clinical and 43 placental phenotypes were statistically compared by ANOVA or Chi-square among 3 groups containing a total of 378 placentas with SFVM: group 1 contained 44 cases of recent SFVM (endothelial fragmentation, villous hypovascularity by CD34 immunostain, and/or stromal vascular karyorrhexis); group 2 contained 264 cases of established SFVM (clusters of avascular villi); and group 3 contained 70 cases of remote SFVM (villous mineralization). Statistically significant differences among the three study groups (p Bonferroni < 0.002) were found in four clinical variables (gestational age, frequencies of macerated stillbirth, induction of labor, and cesarean section) and in five placental variables (frequencies of fetal vascular ectasia, stem vessel luminal vascular abnormalities, diffusely increased extracellular matrix in chorionic villi, chorionic disk extravillous trophoblast microcysts, and excessive extravillous trophoblasts in the chorionic disc). In summary, the absence of statistically significant differences between the study groups regarding the most common causes of SFVM (hypertensive conditions of pregnancy, diabetes mellitus, fetal anomalies, and clinical and pathological features of umbilical cord compromise) is evidence that the three types of SFVM reflect temporal heterogeneity rather than etiopathogenesis. This evidence can be used to date the onset of fetal vascular malperfusion before delivery or stillbirth. The coexistence of different SVFM lesions of various durations indicates ongoing or repeat occurrences of FVM rather than single episodes.


Leveraging high-throughput purification to accelerate viral vector process development.
Recombinant adeno-associated virus (AAV) has been broadly used as a delivery tool for gene therapy applications. The development of a robust purification process is essential for delivering high purity and quality AAV products to clinic. The short clinical timelines and material limitations of early-stage development pose unique challenges to developing robust and scalable downstream purification processes. One approach to overcome these limitations is to leverage high throughput (HTP) strategies and automation technologies for purification process development, an approach that is well established in protein biologics and other areas. However, due to the unique challenges related to viral vector purification, implementing HTP approaches for gene therapy process development has not been explored extensively. In this paper, we established a HTP chromatography platform and demonstrated its capability to facilitate gene therapy purification process development using both mini-columns and self-packed resin plates. The end-to-end development workflow for AAV HTP purification is detailed in this work with the expectation of serving as an introductory for the AAV purification development field. Comparable process performance was confirmed between a bench-scale chromatography process and an HTP chromatography format. Slightly lower recovery was observed using the HTP format (62% vs 75%), as well as %full capsid enrichment (71% vs. 82%). Comparable impurity clearance capability was demonstrated between the two different systems as well. It was concluded that the established HTP chromatography formats can serve as a surrogate to bench-scale chromatography development to reduce material needs and development timelines for AAV purification development.


Novel catalytic oxidative coupling reaction of N,N-dimethyl-p-phenylenediamine with 1,3-phenylenediamine and its applications to the determination of copper and iron at trace levels by flow injection technique.
A new catalytic oxidative coupling reaction of N,N-dimethyl-p-phenylenediamine (DPD) with 1,3-phenylenediamine (mPD) in the presence of hydrogen peroxide has been developed for trace metals analysis. The rate of the oxidation/coupling reaction can be enhanced significantly by iron, copper and cobalt. These metal ions can catalyze the oxidation reaction of DPD to form an oxidized product; the oxidized DPD was then coupled with mPD to give a blue-colored product which was measured spectrophotometrically at 650 nm. On the basis of such a reaction scheme, two simple flow injection analysis methods for the determination of copper and iron have been developed. Detailed studies on chemical and FIA variables affecting the sensitivity of the detection were carried out. Interferences from several ionic species were examined for the determination of copper: the interference effect by Fe(III) and Fe(II) up to 1.5 mg L(-1) was successfully suppressed by pretreating sample with ammonium acetate buffer solution (pH 8.4). Good linearity of a standard calibration graph was obtained over the ranges of 0-8 and 0-2 microg L(-1) of copper and iron, respectively, and the detection limits were 0.05 and 0.02 microg L(-1) for copper and iron, respectively. The precision of the methods in terms of relative standard deviation were 1.4 and 1.5% of R.S.D. which were obtained from 10 injections of 2.0 and 1.0 microg L(-1) of standard copper and iron, respectively. The proposed methods were successfully applied to the determination of copper and iron in tap and river water samples. The accuracy of the proposed methods was assessed by the analysis of certified reference material of river water.


Comparison of clinical characteristics of first-episode thrombotic thrombocytopenic purpura and TTP-like syndrome: a retrospective cohort study in a level I hematology center in China.
Comparing the characteristics of thrombotic thrombocytopenic purpura (TTP) and TTP-like syndrome patients at admission will allow early differentiation of TTP from TTP-like syndrome and help tailor initial treatment. The medical records of 78 patients with suspected TTP in the Emergency Department of Peking University People's Hospital in the past 5 years were retrospectively analyzed and divided into TTP and TTP-like syndrome groups based on ADAMTS13 activity and ADAMTS13 antibody titer. There were 25 and 53 patients in the TTP group and the TTP-like syndrome group, respectively. The neutrophil-to-lymphocyte ratio (P = 0.025) was tremendously higher, and albumin (P = 0.002) was lower in the TTP-like syndrome group, indicating a more severe inflammation. Compared with the TTP-like syndrome group, the TTP group had an approximately two-fold to three-fold higher prevalence of central nervous system dysfunction (P < 0.001). Also, hemolysis was more substantial in the TTP group as evidenced by higher schistocytes (P < 0.001), reticulocyte (P < 0.001), total bilirubin (P = 0.002), indirect bilirubin (P < 0.001), lactate dehydrogenase (P = 0.007) and cell-free hemoglobin (P < 0.001), simultaneously lower platelet (P < 0.001), haptoglobin (P = 0.044), and ADAMTS13 activity (P < 0.001). The Kaplan-Meier survival analysis showed that the TTP group significantly predicted poor prognosis (log-rank test: X2 = 5.368, P = 0.021). TTP and TTP-like syndrome are two kinds of distinct phenotypes with different hemolysis statuses and illustrated differentiated inflammatory reactions, target organ damage (TOD), and the clinical outcome.


Phosphatidylserine externalization in caveolae inhibits Ca2+ efflux through plasma membrane Ca2+-ATPase in ECV304.
It has been evidenced that plasma membrane Ca(2+)-ATPase (PMCA) is localized at caveolae. However, the caveolar function of PMCA in living cells has never been demonstrated. In the present study, PMCA is exclusively localized at caveolae from ECV 304 cells demonstrated by sucrose gradient fractionation and the co-localization of PMCA with caveolin-1 was visualized by confocal microscopy. We found that PMCA is the main mechanism involved in Ca(2+) efflux in ECV 304 cells. Treatment of cells with MbetaCD to disrupt caveolae significantly reduced the Ca(2+) efflux, and the rate of decay is 4.45+/-0.14 min(-1) in the absence of MbetaCD and 1.99+/-0.038 min(-1) in the presence of MbetaCD. Moreover, the replenishment of cholesterol restored the reduction of the PMCA-mediated Ca(2+) efflux in the presence of MbetaCD. Consistent with Ca(2+) efflux in living cells, the activity of the reconstituted PMCA in membranes extracted from cells in vitro was decreased in the presence of MbetaCD. It was found that phosphatidylserine, which is normally in the inner leaflet of plasma membranes and is able to stimulate PMCA was relatively enriched in caveolae. Importantly, the treatment of cells with MbetaCD concomitantly increased the phosphatidylserine externalization. Taken together, our results suggest that activation of PMCA in caveolae is modulated by phosphatidylserine, and phosphatidylserine externalization induced by MbetaCD reduced the interaction of phosphatidylserine with PMCA, subsequently PMCA-mediated Ca(2+) efflux in ECV 304 cells.


Association Between rs217727 and rs2839698 H19 Polymorphisms and Obesity.
Obesity is a worldwide health problem with an increasing trend. This condition has a significant genetic background. H19 lncRNA has been shown to protect from dietary obesity through decreasing levels of monoallelic genes in brown fat. In the current study, we aimed to find the association between two possibly functional H19 polymorphisms, namely rs217727 and rs2839698 and obesity in Iranian population. These polymorphisms have been shown to affect risk of some obesity-related conditions in different populations. The study included 414 obese cases and 392 controls. Notably, both rs2839698 and rs217727 were associated with obesity in the allelic model as well as all supposed inheritance models. In addition, after adjustment for gender, all P values remained significant. For rs2839698, the OR (95% CI) for T allele vs. C allele was 3.29 (2.67-4.05) (P-value < 0.0001). In the co-dominant model, both TT and CT genotypes were found to confer risk of obesity compared with CC genotype (OR (95% CI)= 14.02 (8.39-23.43) and 9.45 (6.36-14.04), respectively). Similarly, combination of TT and CT genotypes had an OR (95% CI) = 10.32 (7.03-15.17) when compared with CC genotype. For rs217727, the T allele was found to exert a protective effect (OR (95% CI) = 0.6 (0.48-0.75)). Moreover, in the co-dominant model, OR (95% CI) values for TT and TC genotypes vs. CC genotype were 0.23 (0.11-0.46) and 0.65 (0.49-0.87), respectively. Taken together, H19 polymorphisms may affect risk of obesity in Iranian population. It is necessary to conduct functional studies to confirm a causal relationship between the rs217727 and rs2839698 polymorphisms and obesity.


Prognostic nutritional index as a risk factor for diabetic kidney disease and mortality in patients with type 2 diabetes mellitus.
Microinflammation and malnutrition are common in individuals with type 2 diabetes mellitus (T2DM). We aimed to validate whether prognostic nutritional index (PNI) may increase the risk of diabetic kidney disease (DKD) and all-cause mortality in T2DM patients. This retrospective cohort study was based on the National Health and Nutrition Examination Survey (NHANES) and National Death Index (NDI) 2013-2018 database. A total of 14,349 eligible subjects were included, and 2720 of them were with T2DM. PNI was assessed by the 5 × lymphocyte count (109/L) + serum albumin (g/L). The Logistic and Cox regression analyses were conducted to investigate the risk factors of DKD and mortality in T2DM patients. For 14,349 participants represented 224.7 million noninstitutionalized residents of the United State, the average PNI was 53.72 ± 0.12, and the prevalence of T2DM was 14.89%. T2DM patients had a lower level of PNI and dietary protein intake, a higher risk of mortality, kidney injury, anemia, arterial hypertension and hyperuricemia, compared with non-T2DM subjects. DKD occurred in 35.06% of diabetic participants and a higher PNI was independently related with a lower risk of DKD (OR 0.64, 95% CI 0.459-0.892, p = 0.01) in T2DM after multivariate adjustment. During a median follow-up of 46 person-months (29-66 months), a total of 233 T2DM individuals died from all causes (mortality rate = 8.17%). Subjects with T2DM who had a higher PNI showed a lower risk of all-cause mortality (HR 0.60, 95% CI 0.37-0.97, p = 0.036). PNI, as a marker of immunonutrition, correlated with the incidence of DKD, and was an independent predictor for all-cause mortality in participants with T2DM. Thus, PNI may conduce to the risk stratification and timely intervention of T2DM patients.


Multivariate analysis of enriched landfill soil consortia provide insight on the community structural perturbation and functioning during low-density polyethylene degradation.
Plastic-enriched sites like landfills have immense potential for discovery of microbial consortia that can efficiently degrade plastics. In this study, we used a combination of culture enrichment, high-throughput PacBio sequencing of 16 S rRNA and the ITS gene, Fourier transform infrared (FTIR), and scanning electron microscopy (SEM) to examine the compositional and diversity perturbations of bacterial and fungal consortia from landfill soils and their impact on low-density polyethylene (LDPE) film biodegradation over a 90-day period. Results showed that enrichment cultures effectively utilized LDPE as a carbon source for cellular growth, resulting in significant weight reduction (22.4% and 55.6%) in the films. SEM analysis revealed marked changes in the micrometric surface characteristics (cracks, fissures, and erosion) and biofilm formation in LDPE films. FTIR analyses suggested structural and functional group modification related to C-H (2831-2943 cm⁻¹), and CH₂ (1400 cm⁻¹) stretching, CO and CC (680-950 cm⁻¹) scission, and CO incorporation (3320-3500 cm⁻¹) into the carbon backbone, indicative of LDPE polymer biodegradation. Enrichment cultures had lower diversity and richness of microbial taxa compared to soil samples, with LDPE as a carbon source having a direct influence on the structure and functioning of the microbial consortia. A total of 26 bacterial and 12 fungal OTU exhibiting high relative abundance and significant associations (IndVal > 0.7, q < 0.05) were identified in the enrichment culture. Bacterial taxa such as unclassified Parvibaculum FJ375498, Achromobacter xylosoxidans, unclassified Chitinophagaceae PAC002331, unclassified Paludisphaera and unclassified Comamonas JX898122, and six fungal species (Galactomyces candidus, Trichosporon chiropterorum, Aspergillus fumigatus, Penicillium chalabudae, Talaromyces thailandensis, and Penicillium citreosulfuratum) were identified as the putative LDPE degraders in the enrichment microbial consortium cultures. PICRUSt2 metagenomic functional profiling of taxonomic bacterial taxa abundances in both landfill soil and enrichment microbial consortia also revealed differential enrichment of energy production, stress tolerance, surface attachment and motility pathways, and xenobiotic degrading enzymes important for biofilm formation and hydrolytic/oxidative LDPE biodegradation. The findings shed light on the composition and structural changes in landfill soil microbial consortia during enrichment with LDPE as a carbon source and suggest novel LDPE-degrading bacterial and fungal taxa that could be explored for management of polyethylene pollution.


Identification of an RNase that preferentially cleaves A/G nucleotides.
Ribonucleases play an important role in the RNA metabolism which is critical for the localization, stability and function of mature RNA transcripts. More and more ribonucleases were discovered in recent years with the progress of technology. In the present study, we found that the uncharacterized C19orf43, a novel interacting protein of human telomerase RNA (hTR), digested T7 transcribed RNA, total cellular RNA and RNA oligos but not DNA. Thus we named this new RNase as hTRIR (human telomerase RNA interacting RNase). Genetic analysis showed that hTRIR is conserved among eukaryotic species and widely expressed in different cell lines. The RNase activity of hTRIR works in a broad temperature and pH range while divalent cations are not required. The conserved C-terminus of C19orf43 is necessary for its activity. Finally, we found that hTRIR cleaves all four unpaired RNA nucleotides from 5' end or 3' end with higher efficiency for purine bases, which suggested that hTRIR is an exoribonuclease. Taken together, our study showed the first evidence of the novel function of hTRIR in vitro, which provides clue to study the regulatory mechanism of hTR homeostasis in vivo.


The synthesis of 64Cu-chelated porphyrin photosensitizers and their tumor-targeting peptide conjugates for the evaluation of target cell uptake and PET image-based pharmacokinetics of targeted photodynamic therapy agents.
Targeted photodynamic therapy (PDT) is necessary for preventing the side effects associated with PDT, such as photosensitivity caused by the distribution of photosensitizers into normal tissues. In the development of targeted PDT agents, a simple evaluation system of in vivo pharmacokinetics, as well as target cell uptake, is absolutely imperative. We hypothesized that (64)Cu chelation with porphyrin photosensitizer-biomacromolecule conjugates may become a simple and versatile labeling strategy for this purpose. Protoporphyrin IX (PPIX) and a bombesin (BBN) analog, that interacts with the gastrin-released peptide (GRP) receptor, were used as a photosensitizer and tumor-targeting peptide, respectively. Then, a conjugate of PPIX and BBN analog linked via short polyethylene glycol (PPIX-PEG6-BBN analog) was synthesized and used as a targeted PDT agent. In addition, a (64)Cu-chelated PPIX-PEG6-BBN analog was synthesized under optimized reaction conditions. Lastly, cell uptake study and PET image-based pharmacokinetic analyses of the PPIX-PEG6-BBN analog were carried out in a human prostate cancer cell line, PC-3, which highly expresses the GRP receptor, and PC-3 tumor-bearing mice. It was confirmed that degradation (thought to be based on radiolysis) occurs, and large amounts of (64)Cu-labeling compounds are wasted in the reaction mixture. Interestingly, the addition of ethanol into the reaction mixture provides an effective solution for this problem. As for cell uptake study, the [(64)Cu]PPIX-PEG6-BBN analog demonstrated significantly higher uptake for PC-3 cells than [(64)Cu]PPIX and, in addition, the uptake of [(64)Cu]PPIX-PEG6-BBN analog was significantly inhibited by adding excess cold BBN analog peptide. PET image-based pharmacokinetic evaluation revealed that [(64)Cu]PPIX-PEG6-BBN analog and [(64)Cu]PPIX rapidly accumulate into the liver and kidney, circulate in blood for a long time compared with normal peptides, and distribute at a low level in the tumor. This result suggested that in vivo biodistribution of PPIX-PEG6-BBN analog is mainly dependent on the lipophilicity of PPIX. Ex vivo measurements of radioactivity distribution after PET studies showed that although there was no remarkable difference in the tumor/skin ratio of radioactivity between [(64)Cu]PPIX-PEG6-BBN analog and [(64)Cu]PPIX, the pancreas (an organ that also expresses GRP receptors)/skin ratio was significantly higher in the case of [(64)Cu]PPIX-PEG6-BBN analog. We report on the successful synthesis of (64)Cu-chelated porphyrin photosensitizers and their tumor-targeting peptide conjugates under conditions in which radiolysis is suppressed. This labeling strategy with porphyrin photosensitizers may be of value for the rapid development of ideal targeted PDT agents.


Intravitreal injection worsens outcomes in a mouse model of indirect traumatic optic neuropathy from closed globe injury.
It is well established that an intravitreal needle poke or injection of buffer is protective to the retina in models of photoreceptor degeneration due to release of endogenous neurotrophic factors. Here we assess the effect of intravitreal injection of buffer in a model of closed globe trauma that causes air-blast induced indirect traumatic optic neuropathy (bITON). We injected animals 1-day after the last bITON or sham procedure and performed assessments 1-month later. Surprisingly, we detected a lower electroretinogram (ERG), greater optic nerve damage, and increased levels of pro-inflammatory cytokines in animals given an intravitreal injection. The effect was sometimes independent of bITON and sometimes exacerbated by the injury. Retina histology appeared normal, however the total number of axons in the optic nerve was lower even in uninjured animals that were injected. The number of degenerative axons was further increased in injured animals that were injected. In contrast, we detected a decrease in the ERG a wave and b wave amplitudes, but no effect on the visual evoked potential. Levels of the pro-inflammatory cytokines, IL-1α and IL-1β were elevated in the mice that received an intravitreal injection. This increase was even greater in animals that also had a bITON. This suggests that intravitreal injections may be injurious to the optic nerve particularly during the acute stage of optic nerve injury. In addition, the data suggests a role for IL-1α and IL-1β in this response.


Risk factors for poor outcome and mortality at 3 months after the ischemic stroke in patients with atrial fibrillation.
It has been shown that patients with atrial fibrillation have a poor prognosis in the early recovery phase after ischemic stroke (IS) or transient ischemic attack (TIA). The purpose of this study was to identify the risk factors associated with poor outcome, including mortality, 3 months after the onset of IS in patients with atrial fibrillation. We have prospectively investigated the characteristics of patients selected from the China National Stroke Registry. Poor outcome was defined as the modified Rankin scale score of 3 or more or death at 3-month follow-up. Association between the relevant risk factors and poor outcome was analyzed using logistic regression analysis. Additionally, the interaction between multiple risk factors was also analyzed. Each year of age (odds ratio [OR]: 1.031; 95% confidence interval [CI] 1.017-1.045), the National Institutes of Health Stroke Scale (NIHSS) at admission (OR: 1.219; 95% CI 1.185-1.254), and female gender (OR: 1.710; 95% CI 1.296-2.256) were independent risk factors for poor outcome at 3 months after IS. Independent risk factors for 3-month mortality included age (OR: 1.024; 95% CI 1.007-1.041), NIHSS at admission (OR: 1.122; 95% CI 1.100-1.144), and history of heart failure (OR: 1.855; 95% CI 1.141-3.015). Conversely, heavy alcohol intake was associated with protective effect on mortality poststroke (OR: .400; 95% CI .173-.928). There was no significant interaction between age and gender (for mortality, P=.16; for poor outcomes, P=.91), age and NIHSS (for mortality, P=.38; for poor outcomes, P=.11), and gender and NIHSS (for mortality, P=.33; for poor outcomes, P=.80). Age, gender, and NIHSS score were independently associated with poor outcome for IS or TIA patients with nonvalvular atrial fibrillation in the early recovery stage.


[Streptococcus pneumoniae-induced hemolytic uremic syndrome: a serotype-3-associated case].
Hemolytic uremic syndrome (HUS) is the primary cause of acute renal failure in children younger than 3 years of age. It usually occurs after a diarrheal illness due to Shiga-toxin-producing Escherichia coli. Streptococcus pneumoniae (SP)-induced HUS remains rare, involving 5% of all cases of HUS in children, but its frequency has increased over the last decade. The incidence of HUS following invasive pneumococcal infections is estimated at 0.4 to 0.6%. We report here the case of a 3.5-year-old child who presented SP serotype-3-associated HUS. The diagnosis was suspected by the patient's multiple organ failure. The pathogenesis involves the activation of the Thomsen-Friedenreich antigen. To prevent transfusion-associated hemolysis, it is recommended that fresh-frozen plasma or unwashed blood products should be avoided when possible. Our patient was transfused with 4 units of unwashed red blood cell and 2 units of fresh-frozen plasma. No special complication was noted. The risk of immediate complications requires close clinical and biological monitoring, and the possibility of starting dialysis immediately. Twenty-five to 35% of SP-HUS patients exhibit long-term renal aftereffects. The acute mortality rate depends on the site of infection. The increased frequency of SP-HUS may be related to the new ecology of serotypes created by widespread Prevenar7(®) vaccination.


[Ways of supporting relatives in intensive care units : Overview and update].
Relatives of patients in the intensive care unit (ICU) face a challenging situation: they often experience an existential crisis with great emotional stress and at the same time they are often actively involved in therapeutic decisions. The visiting restrictions of the coronavirus disease 2019 (COVID-19) pandemic have created new challenges in providing support to relatives. The aim of this work is to present current and new developments in supporting relatives of critically ill patients in the form of a narrative review. In recent years, numerous new approaches and projects to support relatives have been developed. They can be assigned to the following six areas: 1) presence of relatives in the ICU, 2) proactive involvement in care, 3) structured communication/information and online offers, 4) multidisciplinary cooperation, 5) organizational management and 6) follow-up offers. The evidence and the current implementation status of these measures are very heterogeneous internationally and nationally. Measures for providing support for ICU relatives are diverse. Some can even be implemented despite visit bans. Recent digital developments enable virtual visits and a better exchange of information between the ICU team and relatives.


Peptidyl 3-substituted 1-hydroxyureas as isosteric analogues of succinylhydroxamate MMP inhibitors.
To evaluate N-hydroxyurea as zinc binding group in the design of MMP inhibitors, two peptidyl 1-hydroxyureas were prepared by N-hydroxycarbamoylation of the diastereomeric dipeptides H-Leu-Phe-NHMe and H-D-Leu-Phe-NHMe. Peptidyl 1-hydroxyureas were more potent than the parent peptides, but dramatically weaker (4-5 orders of magnitude) than the isosteric (R)-succinylhydroxamate analogue, which displays IC(50) in the range of nM vs MMP-1, -3, -7 and sub-nM vs MMP-2, -8, and -9. The peptidyl 1-hydroxyurea 1a attained an IC(50) of 20 microM vs MMP-9, and substantially approaches inhibition of known N-hydroxyureas based on aminoacids or peptides against other zinc metalloenzymes and non-peptidic N-hydroxyureas against MMPs. Strong preference of the O-N1-C=O unit for the antiperiplanar amide bond conformation seems to be the major limit for more effective zinc chelation. Methylation of a peptidyl 1-hydroxyurea at N3, to promote the synperiplanar O-N1-C=O conformation required for zinc chelation and improve affinity, resulted in release of a methylimidazolidine-2,4-dione through an undesired intramolecular reaction reminiscent of the Edman peptide degradation.


The investigation of unique water-soluble heptamethine cyanine dye for use as NIR photosensitizer in photodynamic therapy of cancer cells.
In this paper, a unique water-soluble heptamethine cyanine dye as NIR photosensitizer was synthesized to explore its properties associated with potential applications in photodynamic therapy (PDT). In the strategy of designing this photosensitizer, a sulfonic acid was used as a water soluble functional group and linked to the fluorophore through alkyl chains. 4-amino-2,2,6,6-tetramethylpiperidine-N-oxyl(Tempo) moiety was used as the a nitroxide spin label in obtaining biochemical reaction information in vivo due to it could greatly increase the inter-system crossing (ISC) process for triplet-state photosensitizers and low toxicity. As expected, the photosensitizers performed well in vitro photodynamic therapy. There were a remarkable absorbance band located at 692 nm and emission peaks falls at 762 nm, the quantum yield (Φf) was calculated to be 12.12% in pure aqueous solution using ICG as standards. The photosensitizer also has high singlet oxygen quantum yield (Φ△) for 16.96% with NIR LED irradiation. This photosensitizer can rapidly produce singlet oxygen and exhibit high phototoxicity under NIR light irradiation. It has excellent cellular uptake ability and better cell compatibility. It was also successfully applied in Near-infrared fluorescence imaging and AO/EB staining. In a whole, the organic dye based on Heptamethine cyanine used as photosensitizer has great potential in vivo cancer treatment.


Impacts of Preoperative Serum Albumin Level on Outcomes of Cytoreductive Surgery and Perioperative Intraperitoneal Chemotherapy.
Only few small studies in the literature have explored the impacts of preoperative serum albumin level and clinical outcomes of patients with peritoneal surface malignancy (PSM) who underwent cytoreductive surgery (CRS) and perioperative intraperitoneal chemotherapy (PIC). This study aimed to evaluate the value of preoperative serum albumin as a prognostic factor for long-term survival outcomes after CRS and PIC in a large patient cohort and to determine whether preoperative serum albumin is correlated with perioperative complications. This retrospective study examined prospectively collected data for patients with PSM who underwent CRS and PIC by one surgical team at St George Hospital in Sydney, Australia. The study used 35 g/L as the cuffoff for normal serum albumin level. The study enrolled 591 patients. Hypoalbuminemia was found to be associated with a significantly higher rate of major morbidity (p < 0.001), a longer ICU stay (p = 0.003), a longer HDU stay (p < 0.001), a longer total hospital stay (p < 0.001), and a shorter overall survival (OS) (p = 0.016). Factor analysis showed preoperative serum hypoalbuminemia to be a prognostic factor for a poor perioperative outcome (p = 0.018) and a poor OS (p = 0.026). Preoperative hypoalbuminemia is associated with poor perioperative outcomes. More importantly, it is a predictor of poorer OS for patients with PSM independent of the PCI, age, and completeness of cytoreduction. In the future, strategies should be undertaken to improve preoperative nutrition of malnourished patients as a means of improving clinical outcomes for patients with PSM.


Congruence as a measurement of extended haplotype structure across the genome.
Historically, extended haplotypes have been defined using only a few data points, such as alleles for several HLA genes in the MHC. High-density SNP data, and the increasing affordability of whole genome SNP typing, creates the opportunity to define higher resolution extended haplotypes. This drives the need for new tools that support quantification and visualization of extended haplotypes as defined by as many as 2000 SNPs. Confronted with high-density SNP data across the major histocompatibility complex (MHC) for 2,300 complete families, compiled by the Type 1 Diabetes Genetics Consortium (T1DGC), we developed software for studying extended haplotypes. The software, called ExHap (Extended Haplotype), uses a similarity measurement we term congruence to identify and quantify long-range allele identity. Using ExHap, we analyzed congruence in both the T1DGC data and family-phased data from the International HapMap Project. Congruent chromosomes from the T1DGC data have between 96.5% and 99.9% allele identity over 1,818 SNPs spanning 2.64 megabases of the MHC (HLA-DRB1 to HLA-A). Thirty-three of 132 DQ-DR-B-A defined haplotype groups have > 50% congruent chromosomes in this region. For example, 92% of chromosomes within the DR3-B8-A1 haplotype are congruent from HLA-DRB1 to HLA-A (99.8% allele identity). We also applied ExHap to all 22 autosomes for both CEU and YRI cohorts from the International HapMap Project, identifying multiple candidate extended haplotypes. Long-range congruence is not unique to the MHC region. Patterns of allele identity on phased chromosomes provide a simple, straightforward approach to visually and quantitatively inspect complex long-range structural patterns in the genome. Such patterns aid the biologist in appreciating genetic similarities and differences across cohorts, and can lead to hypothesis generation for subsequent studies.


Hypericum perforatum L. extract exerts insulinotropic effects and inhibits gluconeogenesis in diabetic rats by regulating AMPK expression and PKCε concentration.
Hypericum perforatum L., commonly known as St. John's Wort (SJW), represents one of the best-known and most thoroughly researched medicinal plant species. The ethnobotanical usage and bioactivities related to H. perforatum include treatment of skin diseases, wounds and burns, gastrointestinal problems, urogenital diseases and psychiatric disorders, particularly depression. In the last decade, many studies focused on the bioactive constituents responsible for the antihyperglycemic and antidiabetic activity of SJW extracts. However, the mechanism by which H. perforatum extract exhibits these properties is still unclear. Hence, the current study was designed to gain insight into the underlying biochemical and molecular mechanisms by which wildly growing H. perforatum exerts its antihyperglycemic and antidiabetic activities. Plant material of H. perforatum was harvested from a natural population in the Republic of North Macedonia during full flowering season. Methanol (80% v/v) was used to extract bioactive components from HH powder. The dissolved HH dry extract (in 0.3% CMC) was given daily as a single treatment (200 mg/kg bw) during 14 days both in healthy and streptozotocin-induced diabetic rats. As a positive control, we applied glibenclamide. The activity of key enzymes involved in carbohydrate methabolisam in the liver were assessed, along with substrate concentration, as well as AMPK mRNA levels, PKCε concentration, plasma insulin level and pancreatic PARP activity. Compared to diabetic rats, treatment of diabetic rats with HH extract resulted with decreased activity of hepatic enzymes glucose-6-phospatase and fructose-1,6-bisphosphatase, increased liver glycogen and glucose-6-phosphate content, which resulted with reduced blood glucose concentration up to normoglycaemia. Non-significant changes were observed in the activity of hexokinase, glycogen phosphorylase and glucose-6-phospahte dehydrogenase. HH-treatment also caused an increase in plasma insulin concentration and increase in pancreatic PARP activity. Finally, HH treatment of diabetic rats showed significant increase in AMPK expression and decrease of PKCε concentration. We present in vivo evidence that HH- extract exert insulinotropic effects and regulate endogenous glucose production mostly by suppressing liver gluconeogenesis. The HH-treatment did not effected glycogenolysys and glycolysis. Finally, we confirm the antihyperglycemic and antidiabetic effect of HH-extract and the mechanism of this effect involves amelioration of AMPK and PKCε changes in the liver.


Construction of polycistronic baculovirus surface display vectors to express the PCV2 Cap(d41) protein and analysis of its immunogenicity in mice and swine.
To increase expression levels of the PCV2 Cap(d41) protein, novel baculovirus surface display vectors with multiple expression cassettes were constructed to create recombinant baculoviruses BacSC-Cap(d41), BacDD-2Cap(d41), BacDD-3Cap(d41), and BacDD-4Cap(d41). Our results reveal that the recombinant baculovirus BacDD-4Cap(d41) was able to express the highest levels of Cap(d41) protein. Optimum conditions for expressing the PCV2 Cap(d41) protein were determined, and our results show that 107 of Sf-9 infected with the recombinant baculovirus BacDD-4Cap(d41) at an MOI of 5 for 3 days showed the highest level of protein expression. Mice immunized with the 4Cap(d41) vaccine which was prepared from the recombinant baculovirus-infected cells (107) elicited higher ELISA titers compared to the Cap (d41) vaccine. The 4Cap(d41) vaccine could elicit anti-PCV2 neutralizing antibodies and IFN-γ in mice, as confirmed by virus neutralization test and IFN-γ ELISA. Moreover, the swine lymphocyte proliferative responses indicated that the 4Cap(d41) vaccine was able to induce a clear cellular immune response. Flow cytometry analysis showed that the percentage of CD4+ T cells and CD4+/CD8+ ratio was increased significantly in SPF pigs immunized with the 4Cap(d41) vaccine. Importantly, the 4Cap(d41) vaccine induced an IFN-γ response, further confirming that its effect is through cellular immunity in SPF pigs. An in vivo challenge study revealed that the 4Cap(d41) and the commercial vaccine groups significantly reduce the viral load of vaccinated pigs as compared with the CE negative control group. Taken together, we have successfully developed a 4Cap(d41) vaccine that may be a potential subunit vaccine for preventing the disease associated with PCV2 infections.


Determination of biomarkers for polycyclic aromatic hydrocarbons (PAHs) toxicity to earthworm (Eisenia fetida).
Polycyclic aromatic hydrocarbon (PAH) compounds are persistent, carcinogenic, and mutagenic. When PAHs enter agricultural soils through sewage sludge, they pose an environmental risk to soil organisms, including earthworms. Therefore, we aimed to determine the toxic effects of PAHs on earthworms. Five PAHs were used: fluorene, anthracene, phenanthrene, fluoranthene, and pyrene. Only fluorene and phenanthrene exhibited toxicity (LC50 values 394.09 and 114.02 g L(-1), respectively) against the earthworm Eisenia fetida. None of the other PAHs tested in this study enhanced the mortality of adult earthworm until the concentrations reached to 1000 g L(-1). After exposure to PAHs, acetylcholinesterase (AChE) activity in E. fetida decreased in a concentration-dependent manner, and phenanthrene exhibited the strongest inhibitory effect on AChE, followed by fluorene. Activity of a representative detoxifying enzyme, carboxylesterase, was dramatically reduced in E. fetida exposed to all tested PAHs in comparison with that observed in the control test. The remaining glutathione S-transferase activity significantly decreased in E. fetida after exposure to PAHs. To profile small proteins <20 kDa, SELDI-TOF MS with Q10 ProteinChips was used, and 54 proteins were identified as being significantly different from the control (p = 0.05). Among them, the expressions of three proteins at 4501.8, 4712.4, and 4747.9 m/z were only enhanced in E. fetida exposed to anthracene and pyrene. One protein with 16,174 m/z was selectively expressed in E. fetida exposed to fluorene, phenanthrene, and fluoranthene. These proteins may be potential biomarkers for the five PAHs tested in E. fetida.


Progression toward metastatic phenotype with loss of growth-inhibiting tumor-cell/cell interactions in vivo.
Five autonomous sublines, T4-O1320, T4-O1320CY, T4-O1165, T4-O1145 and T4-O196, were established from the transplantable hormone-dependent mouse mammary tumor, TPDMT-4, by passaging under different conditions. These autonomous tumors were characterized by rapid growth in DDD virgin mice and the parental TPDMT-4 by no growth in these mice. Thus, 10(5) T4-O1320, 2 x 10(4) T4-O1320CY, 2 x 10(3) T4-O1165, 2 x 10(3) T4-O1145 and 10(3) T4-O196 cells were co-injected with 5 x 10(5) TPDMT-4 cells into virgin mice to determine whether or not hormone-dependent tumor cells influence the growth of autonomous tumor cells. TPDMT-4 cells retarded the growth of T4-O1320 and T4-O1320CY tumors but accelerated that of T4-O1165, T4-O1145 and T4-O196. Irradiated TPDMT-4 cells stimulated the growth of all the sublines except T4-O1320. In 3-dimensional collagen-gel culture, T4-O1320 and T4-O1320CY cells formed branched or stellate structures similar to normal mammary glands, as did TPDMT-4, but T4-O1165, T4-O1145 and T4-O196 cells grew as rounded masses with knobs and showed a completely different morphology. T4-O1165, T4-O1145 and T4-O196 cells, but not the others, had lung-colonizing ability. Chromosomal aberration was found in T4-O1320CY and T4-O196 but not in the others. Thus, the susceptibility of autonomous subline tumor cells to growth-inhibitory regulation from the parental hormone-dependent tumor cells correlated well with growth morphology within collagen gels and metastatic ability, but not with chromosomal aberration. The results suggest that metastatically-competent tumor-cell variants, once they appear, may have a growth advantage in hormone-dependent mammary tumors.


Feedback on and knowledge, attitude, and skills at the end of pharmacology practical sessions.
Concern has been raised about inadequate pharmacology teaching in medical schools and the high incidence of prescribing errors by doctors in training. Modifications in pharmacology teaching have been carried out in many countries. The present study was carried out using a semi-structured questionnaire to obtain students' perceptions of their knowledge, attitudes, and skills with regard to different subject areas related to rational prescribing at the end of two-year activity-based pharmacology practical learning sessions in a private medical school in Nepal. The effectiveness of the sessions and strengths and suggestions to further improve the sessions were also obtained. The median total knowledge, attitude, skills and overall scores were calculated and compared among different subgroups of respondents. The median effectiveness score was also calculated. Eighty of the 100 students participated; 37 were male and 43 female. The median knowledge, attitude, and skills scores were 24, 39, and 23, respectively (maximum scores being 27, 45, and 36). The median total score was 86 (maximum score being 108). The effectiveness score for most subject areas was 3 (maximum 4). The strengths were the activity-based nature of the session, use of videos and role-plays, and repeated practice. Students wanted more sessions and practice in certain areas. They also wanted more resources and an internet connection in the practical room. The skills scores were relatively low. The immediate impact of the sessions was positive. Studies may be needed to assess the long term impact. Similar programs should be considered in other medical schools in Nepal and other developing countries.


Roll-out of a nurse-led welfare benefits screening service throughout the largest Local Health Care Co-operative in Glasgow: an evaluation study.
To evaluate the roll-out of a nurse-led Attendance Allowance (AA) screening programme in 24 general practices located within the largest Local Health Care Co-operative in Glasgow. Evaluation study Six hundred and thirty participants aged over 64 years who, in the nurses' clinical judgement, appeared to have care needs were recruited opportunistically by community nurses over a 15-month period. A money advice worker contacted all potential underclaimers offering a home visit to assess for unclaimed benefits. The main outcome measured was the total amount of unclaimed AA, linked benefits and grants. Three hundred and sixty-three participants and 13 relatives were awarded a total of 1,136,424.10 pounds. Of this, 1,016,908.70 pounds was on a recurrent annual basis and 119,515.44 pounds was awarded as lump sums. This method of benefits assessment (community-nurse-led pre-AA screening followed by a home visit from a money advice worker) would appear to be an efficient and effective method of income maximization that could be rolled out nationally within primary care settings located in deprived areas.


Single and joint associations of exposure to polycyclic aromatic hydrocarbons with blood coagulation function during pregnancy: A cross-sectional study.
Association linking polycyclic aromatic hydrocarbons (PAHs) to blood coagulation function during pregnancy remains absent. Hence, we conducted a cross-sectional study including 679 late pregnant women (27.2 ± 5.1 years old) drawn from Zunyi birth cohort, Southwest China. During late pregnancy, ten urinary PAHs metabolites and four clinical blood coagulation parameters were measured, including activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT), and fibrinogen (FIB). Multiple linear regression, Restricted cubic spline (RCS) regression, Bayesian kernel machine regression (BKMR), and quantile g-computation (Q-g) regression were used to investigate their single, nonlinear, and mixed associations. Each 2.7-fold increment in 2-hydroxyfluorene (2-OHFlu), 9-hydroxyfluorene (9-OHFlu), 1-hydroxyphenanthrene (1-OHPhe), 2-hydroxyphenanthrene (2-OHPhe), and 3-hydroxyphenanthrene (3-OHPhe) were associated with 0.287 s, 0.190 s, 0.487 s, and 0.396 s shorter APTT, respectively; each 2.7-fold increment in 2-OHPhe was associated with a 0.047 s longer PT; each 2.7-fold increment in 9-hydroxyphenanthrene (9-OHPhe) and 1-hydroxypyrene (1-OHPyr) were associated with 0.087 s and 0.031 s shorter TT, respectively; and each 2.7-fold increment in 1-hydroxynaphthalene (1-OHNap) was associated with 0.032 g/L higher FIB level. The nonlinear association of 2-OHPhe with APTT and 1-OHNap with FIB were also observed. Furthermore, the shortened APTT and TT associated with PAHs mixture were indicated by BKMR and Q-g model. BKMR also revealed a nonlinear association of 2-OHPhe with PT and an interaction effect of 2-OHPhe and 3-OHPhe on APTT. Our results indicate that urinary PAHs was associated with shortened coagulation time and increased FIB. Therefore, more attention should be paid for late pregnant women to prevent PAHs-associated risk of thrombosis. Future perspective studies to confirm our findings and explore the underlying biological mechanism are warranted.