[Value of contrast-enhanced ultrasonography in detection of endoleak after endovascular repair of infrarenal abdominal aortic aneurysm].
Objective: To investigate the value of contrast-enhanced ultrasound in detecting endoleak after endovascular repair of infrarenal abdominal aortic aneurysm (EVAR). Methods: The postoperative follow-up data of 102 patients with infrarenal abdominal aortic aneurysm treated with EVAR in Tianjin Medical University General Hospital from August 2015 to December 2021 were retrospectively analyzed. There were 79 males and 23 females, aged 50-91 (69.6±7.6) years old. Using CT angiography (CTA) as the gold standard for diagnosing endoleaks, the effectiveness of contrast-enhanced ultrasound and CDUS in detecting endoleaks was evaluated by paired design chi-square test, and the Kappa value was calculated for consistency test. Patients were divided into groups according to body mass index (BMI), and the number of false-negative endoleaks detected by contrast-enhanced ultrasound in each group was calculated, and its ratio to the actual number of endoleaks was calculated to evaluate whether BMI was related to false-negative ultrasound-enhanced ultrasound. Results: A total of 203 follow-up visits met the inclusion criteria. Endoleaks were detected 36 times (17.7%) by CTA, 31 times (15.3%) by contrast-enhanced ultrasound, 16 times (7.9%) by CDUS, and they all detected type Ⅰ, type Ⅱ and type Ⅲ endoleaks. There was no significant difference between contrast-enhanced ultrasound and CTA in endoleak detection rate and determination of endoleak types (endoleak detection rate: 15.3% vs 17.7%; determination of endoleak types: type Ⅰ 4 vs 4, type Ⅱ 26 vs 31, type Ⅲ 1 vs 1; all P>0.05). CDUS and CTA had statistically significant differences in the detection rate of endoleaks and determination of endoleak types (endoleak detection rate: 7.9% vs 17.7%; determination of endoleak types: type Ⅰ 4 vs 4, type Ⅱ 11 vs 31, type Ⅲ 1 vs 1; all P<0.001). Compared with CTA, contrast-enhanced ultrasound has a sensitivity of 83.3%, a specificity of 99.4%, a Youden index of 0.827, a coincidence rate of 96.6%, a positive predictive value of 96.8%, a negative predictive value of 96.5%, and a Kappa value of 0.875(P<0.001). The two showed excellent diagnostic consistency. All 6 endoleaks not detected by contrast-enhanced ultrasound were type Ⅱ endoleak that did not require treatment, and 3(15.8%) occurred in obese patients with a BMI≥32 kg/m2. Compared with CTA, CDUS had a sensitivity of 38.9%, a specificity of 98.8%, a Youden index of 0.377, a coincidence rate of 88.2%, a positive predictive value of 87.5%, a negative predictive value of 88.2%, and a Kappa value of 0.482 (P<0.001). The two showed moderate diagnostic agreement. The correlation coefficient of the maximum diameter of aneurysms measured by ultrasound and CT was r=0.873(P<0.001). Conclusions: Contrast-enhanced ultrasound is accurate in detecting endoleak after infrarenal EVAR, and its sensitivity to endoleak detection in obese patients with BMI≥32 kg/m2 will be reduced. CDUS is not suitable for detection of endoleak after infrarenal EVAR, but it can be used to monitor the change of the largest diameter of aneurysm after EVAR.


Identification of pyramidal cells as the critical elements in hippocampal neuronal plasticity during learning.
The activity of single neurons recorded from rabbit hippocampus during classical conditioning of the nictitating membrane reflex was studied. All cells were first categorized according to their responses after fornix stimulation--i.i., antidromic activation, orthodromic activation, or no activation. The majority of cells that were antidromically activated--pyramidal cells--showed a highly positive correlation between the pattern of unit discharge and the topography of the nicititating membrane response within trial periods. Units that were orthodromically driven by fornix stimulation tended to inhibit during the presentation of trial stimuli, whereas most non-activated cells maintained low spontaneous levels of activity at all times. Thus, the major output neurons of the hippocampus appear to be the neuroanatomical substrate for the large and rapidly developing neuronal plasticity induced by this classical conditioning paradigm.


Nucleosome positioning from tiling microarray data.
The packaging of DNA around nucleosomes in eukaryotic cells plays a crucial role in regulation of gene expression, and other DNA-related processes. To better understand the regulatory role of nucleosomes, it is important to pinpoint their position in a high (5-10 bp) resolution. Toward this end, several recent works used dense tiling arrays to map nucleosomes in a high-throughput manner. These data were then parsed and hand-curated, and the positions of nucleosomes were assessed. In this manuscript, we present a fully automated algorithm to analyze such data and predict the exact location of nucleosomes. We introduce a method, based on a probabilistic graphical model, to increase the resolution of our predictions even beyond that of the microarray used. We show how to build such a model and how to compile it into a simple Hidden Markov Model, allowing for a fast and accurate inference of nucleosome positions. We applied our model to nucleosomal data from mid-log yeast cells reported by Yuan et al. and compared our predictions to those of the original paper; to a more recent method that uses five times denser tiling arrays as explained by Lee et al.; and to a curated set of literature-based nucleosome positions. Our results suggest that by applying our algorithm to the same data used by Yuan et al. our fully automated model traced 13% more nucleosomes, and increased the overall accuracy by about 20%. We believe that such an improvement opens the way for a better understanding of the regulatory mechanisms controlling gene expression, and how they are encoded in the DNA.


Imidacloprid is degraded by CYP353D1v2, a cytochrome P450 overexpressed in a resistant strain of Laodelphax striatellus.
Cytochrome P450s are associated with the metabolising of a wide range of compounds, including insecticides. CYP353D1v2 has been found to be overexpressed in an imidacloprid-resistant strain of Laodelphax striatellus. Thus, this study was conducted to express CYP353D1v2 in Sf9 cells as a recombinant protein, to assess its ability to metabolise imidacloprid. Western blot and carbon monoxide difference spectrum analysis indicated that the intact CYP353D1v2 protein had been successfully expressed in Sf9 insect cells. Catalytic activity tests with four traditional P450-activity-probing substrates found that the expressed CYP353D1v2 preferentially metabolised p-nitroanisole, ethoxycoumarin and ethoxyresorufin with specific activities of 32.70, 0.317 and 1.22 pmol min-1 pmol-1 protein respectively, but no activity to luciferin-H EGE. The enzyme activity for degrading imidacloprid was tested by measuring substrate depletion and formation of the metabolite. Kinetic parameters for imidacloprid were Km 5.99 ± 0.95 µm and kcat 0.03 ± 0.0004 min-1 . The chromatogram analysis showed clearly the NADPH-dependent depletion of imidacloprid and the formation of an unknown metabolite. The UPLC-MS mass spectrum demonstrated that the metabolite was an oxidative product of imidacloprid, 5-hydroxy-imidacloprid. These results suggest that CYP353D1v2 in L. striatellus is capable of degrading imidacloprid, and that enzyme activity can be evaluated well only by some traditional probing substrates. © 2017 Society of Chemical Industry.


Nonlinear Optical Phenomena in a Silicon-Smectic A Liquid Crystal (SALC) Waveguide.
Liquid crystals (LCs) are organic materials characterized by the intermediate properties between those of an isotropic liquid and a crystal with a long range order. The LCs possess strong anisotropy of their optical and electro-optical properties. In particular, LCs possess strong optical nonlinearity. LCs are compatible with silicon-based technologies. Due to these unique properties, LCs are promising candidates for the development of novel integrated devices for telecommunications and sensing. Nematic liquid crystals (NLCs) are mostly used and studied. Smectic A liquid crystals (SALCs) have a higher degree of long range order forming a layered structure. As a result, they have lower scattering losses, specific mechanisms of optical nonlinearity related to the smectic layer displacement without the mass density change, and they can be used in nonlinear optical applications. We theoretically studied the nonlinear optical phenomena in a silicon-SALC waveguide. We have shown theoretically that the stimulated light scattering (SLS) and cross-phase modulation (XPM) caused by SALC nonlinearity can occur in the silicon-SALC waveguide. We evaluated the smectic layer displacement, the SALC hydrodynamic velocity, and the slowly varying amplitudes (SVAs) of the interfering optical waves.


Spinal cord stimulation inhibits cortical somatosensory evoked potentials significantly stronger than transcutaneous electrical nerve stimulation.
Despite the good clinical results elicited by spinal cord stimulation (SCS), the physiological basis of action of SCS is widely unknown. Inhibition of somatosensory evoked potential (SEP) amplitudes by SCS has been described, but it is unclear whether this displays dose dependency. Moreover, it is unknown whether the pain-relieving effect elicited by SCS correlates with the inhibition of SEPs. Finally, this study aimed to answer the question whether there is a difference in the effect on SEPs between SCS and transcutaneous electrical nerve stimulation (TENS), thus between central nervous system stimulation and peripheral nervous system stimulation. Ten patients (4 men and 6 women, age range 40-77 years) with neuropathic lower limb pain were included in the study. All patients had implanted SCS systems with percutaneous type electrodes. Cortical SEPs under SCS and TENS were measured without stimulation, under stimulation at perception threshold (PT), and at maximal threshold (MT) in a crossover design. Cortical SEP amplitudes were significantly inhibited by SCS. Stimulation at PT and at MT both led to a statistically significant inhibition of the SEP amplitude. The difference between amplitude reduction at PT and MT showed a tendency towards significance. The degree of SEP amplitude inhibition did not correlate with pain relief. Inhibition of SEP amplitudes by TENS was weaker than that elicited by SCS. The average percentage of amplitude reduction at MT was twice as high under SCS as it was under TENS. No effects on SEP latencies were seen. SCS exerts a significantly stronger inhibition of SEP amplitudes than TENS. The data hint at a dose dependency of SCS-induced SEP amplitude inhibition. No correlation between SEP amplitude inhibition and pain relief was found.


[Petrous bone cholesteatoma: surgery approach and outcomes].
Objective: To investigate the choice of surgical approach of petrous bone cholesteatoma (PBC)and surgical outcomes. Methods: A retrospective study was performed on 90 patients diagnosed and treated for PBC from January 2000 to December 2014 by the Chinese People's Liberation Army General Hospital otolaryngologists. According to Sanna's classification, 40 out of the 90 cases were supralabyrinthine, five infralabyrinthine, four infralabyrinthine-apical, 25 massive and 16 apical. Five cases underwent transmastoid and retrolabyrinthine approach, translabyrinthine approach was performed on six patients, 19 cases underwent subtotal petrosectomy, seven cases underwent transotic approach, 41 cases underwent middle fossa approach, combined transmastoid/middle fossa approach was performed on 11 cases, translabyrinthine and sphenoid sinus approach were performed on one case. Supralabyrinthine cases mainly applied middle fossa approach (77.5%, 31/40) and combined transmastoid and middle-fossa approach(20.0%, 8/40). Combined transmastoid-retrolabyrinthine approach were applied for all the infralabyrinthine cases (100.0%, 5/5). Infralabyrinthine-apical cases mainly applied subtotal petrosectomy (75.0%, 3/4). Massive cases mainly applied subtotal petrosectomy (60.0%, 15/25), transcochlear approach (20.0%, 5/25), and translabyrinthine approach (16.0%, 4/25). Apical cases mainly applied middle fossa approach (62.5%, 10/16). Results: Ninty percent (18/20) of the patients who had preoperative grade Ⅰ facial nerve function maintained in the postoperative period. Out of 90 cases, only 11 cases received open cavity, and the rest cases received cavityobliteration. There were three cases of recurrence, four cases of cavity infection, three cases of cerebrospinal fluid leakage, and one case of epidural hematoma, who all received surgeries. Conclusions: Sanna's classification should be used to classify different kinds of PBC cases, choose the best surgical approach for different cases, and preserve or repair facial function during removal of PBC, and thus reduce recurrence and complications.


Long-term outcomes after surgical resection of pulmonary metastases from colorectal cancer.
Surgical resection has been widely performed on patients with pulmonary metastases from colorectal cancer with favorable outcomes. However, there are currently no standard surgical indications for pulmonary metastases. We reviewed 94 patients who underwent complete resection of pulmonary metastases from colorectal cancer between November 1991 and April 2013. The cumulative survival rate after pulmonary metastasectomy was calculated, and prognostic factors for long-term survival were analyzed. There were 60 men and 34 women, and their median age was 66 years. The 5-year survival rate was 45.5% after pulmonary metastasectomy. The 5-year survival of patients with colon and rectal cancers was 62.4% and 33.8%, respectively (p = 0.030), and the 5-year survival of those with normal and high carcinoembryonic antigen (CEA) levels before pulmonary resection was 57.0% and 30.9%, respectively (p = 0.038). Multivariate analysis revealed the preoperative CEA level was an independent prognostic factor. Recurrence was identified in 65 of the 94 patients (69.1%) after pulmonary metastasectomy, and the patients who underwent surgical resection for recurrent lesions in the liver or lungs, or both, had better survival than those who received other treatments or palliative care. Surgical resection offers a chance to prolong survival in colorectal cancer patients with resectable pulmonary metastases. Owing to the high recurrence rate, careful postoperative follow-up for early detection is recommended, and even for recurrence, surgical resection should be considered for better survival if the lesions are limited to the liver or lungs, or both.


Mediterranean diet and risk of rheumatoid arthritis: a population-based case-control study.
The Mediterranean diet has been associated with lower mortality and lower risk of cardiovascular diseases and cancer. Although its components have been analysed in several studies, only one study has specifically investigated the association between Mediterranean diet and risk of rheumatoid arthritis (RA), and reported no association. Data on 1721 patients with incident RA (cases) and 3667 controls, matched on age, gender and residential area, from the Swedish epidemiological investigation of RA (EIRA), a population-based case-control study, were analysed using conditional logistic regression. The Mediterranean diet score, ranging from 0 to 9, was calculated from a 124-item food frequency questionnaire. In the EIRA study (median age of participants 53 years), 24.1% of the patients and 28.2% of the controls had high adherence to the Mediterranean diet (a score between 6 and 9). After adjustments for body mass index, educational level, physical activity, use of dietary supplements, energy intake, and smoking, high adherence reduced the odds of developing RA by 21% (OR 0.79; 95% CI 0.65-0.96) as compared to low adherence (a score between 0 and 2). The OR was even lower among men (OR 0.49; 95% CI 0.33-0.73), but no significant association was found among women (OR 0.94; 95% CI 0.74-1.18). An association between high diet score and low risk of RA was observed in rheumatoid factor (RF)-positive (OR 0.69; 95% CI 0.54-0.88), but not RF-negative RA (OR 0.96; 95% CI 0.68-1.34), and in RA characterised by presence of antibodies to citrullinated peptides (ACPA), but not in ACPA-negative RA. In this large population-based case-control study, the Mediterranean diet score was inversely associated with risk of RA. However, an association was only found among men and only in seropositive RA.


Chronic obstructive pulmonary disease mortality in railroad workers.
There is little information describing the risk of non-malignant respiratory disease and occupational exposure to diesel exhaust. US railroad workers have been exposed to diesel exhaust since diesel locomotives were introduced after World War II. In a retrospective cohort study we examined the association of chronic obstructive pulmonary disease (COPD) mortality with years of work in diesel-exposed jobs. To examine the possible confounding effects of smoking, multiple imputation was used to model smoking history. A Cox proportional hazards model was used to estimate an incidence rate ratio, adjusted for age, calendar year, and length of follow-up after leaving work (to reduce bias due to a healthy worker survivor effect). Workers in jobs with diesel exhaust exposure had an increased risk of COPD mortality relative to those in unexposed jobs. Workers hired after the introduction of diesel locomotives had a 2.5% increase in COPD mortality risk for each additional year of work in a diesel-exposed job. This risk was only slightly attenuated after adjustment for imputed smoking history. These results support an association between occupational exposure to diesel exhaust and COPD mortality.


Repellency of aromatic medicinal plant extracts and a steam distillate to Aedes aegypti.
The repellent activity of methanol extracts from 23 aromatic medicinal plant species and a steam distillate against female blood-starved Aedes aegypti was examined in the laboratory by skin test and compared with that of N,N-diethyl-m-toluamide (deet). Responses varied according to plant species. At a dose of 0.1 mg/cm2, the repellency of extracts of Cinnamomum cassia bark (91%), Nardostachys chinensis rhizome (81%), Paeonia suffruticosa root bark (80%), and Cinnamomum camphora steam distillate (94%) was comparable to deet (82%). The duration of the effectiveness for extracts from C. cassia bark and N. chinensis rhizome was comparable to deet and lasted for approximately 1 h. Relatively short duration of repellency was observed in P. suffruticosa root bark extract and C. camphora steam distillate. The plants described merit further study as potential mosquito repellent agents.


Ectopic gastrointestinal variceal bleeding with portal hypertension.
Massive gastrointestinal bleeding from gastrointestinal varices is one of the most serious complications in patients with portal hypertension. However, if no bleeding point can be detected by endoscopy in the predilection sites of gastrointestinal varices, such as the esophagus and stomach, ectopic gastrointestinal variceal bleeding should be considered as a differential diagnosis. Herein, we report a case of ectopic ileal variceal bleeding in a 57-year-old woman, which was successfully diagnosed by multi-detector row CT (MDCT) and angiography and treated by segmental ileum resection. To date, there have been no consensus for the treatment of ectopic ileal variceal bleeding. This review was designed to clarify the clinical characteristics of patients with ectopic ileal variceal and discuss possible treatment strategies. From the PubMed database and our own database, we reviewed 21 consecutive cases of ileal variceal bleeding diagnosed from 1982 to 2017. MDCT and angiography is useful for the rapid examination and surgical resection of an affected lesion and is a safe and effective treatment strategy to avoid further bleeding.


Pharmacological characterization of human and murine neuropeptide s receptor variants.
We have recently shown that Neuropeptide S (NPS) can promote arousal and induce anxiolytic-like effects after central administration in rodents. Another study reported a number of natural polymorphisms in the human NPS receptor gene. Some of these polymorphisms were associated with increased risk of asthma and possibly other forms of atopic diseases, but the physiological consequences of the mutations remain unclear. One of the polymorphisms produces an Asn-Ile exchange in the first extracellular loop of the receptor protein, and a C-terminal splice variant of the NPS receptor was found overexpressed in human asthmatic airway tissue. We sought to study the pharmacology of the human receptor variants in comparison with the murine receptor protein. Here, we report that the N107I polymorphism in the human NPS receptor results in a gain-of-function characterized by an increase in agonist potency without changing binding affinity in NPSR Ile107. In contrast, the C-terminal splice variant of the human NPS receptor shows a pharmacological profile similar to NPSR Asn107. The mouse NPS receptor, which also carries an Ile residue at position 107, displays an intermediate pharmacological profile. Structure-activity relationship studies show that the amino terminus of NPS is critical for receptor activation. The altered pharmacology of the Ile107 isoform of the human NPS receptor implies a mechanism of enhanced NPS signaling that might have physiological significance for brain function as well as peripheral tissues that express NPS receptors.


Deficiency of pigment epithelium-derived factor in nasopharyngeal carcinoma cells triggers the epithelial-mesenchymal transition and metastasis.
Distant metastasis is the primary cause of nasopharyngeal carcinoma (NPC) treatment failure while epithelial-mesenchymal transition (EMT) is the critical process of NPC invasion and metastasis. However, tumor-suppressor genes involved in the EMT and metastasis of NPC have not been explored clearly compared with the oncogenes. In the present study, the expression of pigment epithelium-derived factor (PEDF), a potent endogenous antitumor factor, was diminished in human NPC tissues and associated with clinicopathological and EMT features. The knockdown of PEDF induced EMT in lower metastatic NPC cell lines and overexpression of PEDF restored epithelial phenotype in higher metastatic NPC cell lines with typical EMT. The inhibition of PEDF mediated NPC cell spontaneous metastasis in vivo. LRP6/GSK3β/β-catenin signal pathway rather than AKT/GSK3β pathway was involved in the effects of PEDF on EMT. The expression of PEDF was directly downregulated by elevated miR-320c in NPC. In conclusion, our findings indicate for the first time that PEDF functions as tumor-suppressor gene in the occurrence of EMT and metastasis in NPC. PEDF could serve as a promising candidate for NPC diagnosis, prognosis and treatment.


Myocardial perfusion at rest in uncomplicated type 2 diabetes patients without coronary artery disease evaluated by 320-multidetector computed tomography: A pilot study.
Using computed tomography myocardial perfusion imaging (CTP) to investigate resting myocardial perfusion alterations in uncomplicated type 2 diabetes mellitus (T2DM) patients without obstructive coronary artery disease (CAD).A total of 34 participants with 544 myocardial segments were included prospectively: 17 uncomplicated T2DM patients with no significant coronary artery stenosis on coronary computed tomography angiography and 17 healthy controls. Myocardial perfusion was evaluated by transmural perfusion ratio (TPR). Parameters of cardiac structure and function were measured for cardiac comprehensive assessment. Analyses included descriptive statistics and group comparisons.TPR of segments 5, 7, 9, 10 to 14 were significantly reduced in T2DM group compared with controls (P < .05). When 16 myocardial segments were localized into different areas according to the wall orientations, axial levels of left ventricle and coronary artery territories, respectively, TPR of each area in T2DM group were significantly lower than those in the control group (P < .05). No significant differences were found in cardiac anatomy and function analyses between 2 groups.In uncomplicated T2DM patients without obstructive CAD, myocardial perfusion impairments were present and may develop prior to cardiac morphological and functional abnormalities, which can be early detected by CTP.


Molecular diagnosis of hereditary deafness and application of stepwise testing strategy.
Hereditary deafness is one of the most common sensory disorders in humans, and exhibits high genetic heterogeneity. At present, the commonly used molecular diagnostic methods include gene chip, Sanger sequencing, targeted enrichment sequencing, and whole-exome sequencing, with diagnosis rates reaching 33.5%-56.67%. However, there are still a considerable number of patients who can not get a timely and definitive molecular diagnosis. Furthermore, considering the economic burden on patients' families and the relatively high cost of whole-exome or whole-genome sequencing, it is vital to provide stepwise strategies combining multiple detection methods according to the phenotypes of patients. In this review, we evaluate and discuss the utility of molecular diagnosis and the application of stepwise testing strategies in hereditary deafness to provide reference for the selection of diagnostic strategies.


Anatomical heterogeneity of parathyroid glands in posttransplant hyperparathyroidism.
Successful renal transplantation may be complicated by persistent hyperparathyroidism due to diffuse parathyroid hyperplasia remaining from a prolonged period of pretransplant chronic renal failure treatment. Posttransplant hyperparathyroidism is distinct from primary hyperparathyroidism, being characterized by multiple gland involvement and diffuse hyperplasia rather than a single adenoma. The gross pathologic anatomy of the parathyroid glands was assessed in 17 successful renal transplant recipients. Individual and total gland volumes were measured at the time of total parathyroidectomy and forearm reimplantation. Parathyroid hyperplasia was heterogenous in both location and gland size. Right-sided glands were enlarged more than left-sided ones. Subjects with primary tubulointerstitial disease exhibited greater hyperplasia than patients with glomerular disorders. Clinicians should be aware of the heterogeneity of the gland enlargement in patients with diffuse parathyroid hyperplasia, so that these patients are not misdiagnosed as suffering from adenomatous parathyroid disease.


In vitro effects of preserved or preservative-free antiglaucoma medications on human complement system.
Antiglaucoma drugs have been associated with conjunctival and trabecular inflammatory cell infiltrates. However, the underlying mechanisms are still poorly understood. The aim of this study was to assess the effects of antiglaucoma medications on the complement system, an early mediator of the inflammatory response. Human serum was first treated with a classical or alternative pathway activator (aggregated human IgG or zymosan, respectively) in the presence or the absence of preservative-free or benzalkonium (BAK)-preserved antiglaucoma drugs. CH50 assay was then performed to assess the functional activity of residual complement in treated serum. In the absence of complement activator, the antiglaucoma drugs tested in this study were all devoid of intrinsic complement-activating potency. Preserved and preservative-free carteolol as well as preserved latanoprost did not worsen or prevent complement activation induced by zymosan or aggregated IgG. Unexpectedly, both preserved and unpreserved timolol and betaxolol significantly counteracted the effects of complement activators. Timolol prevented activation triggered by both IgG and zymosan to the same extent (24% to 29%), despite the presence of BAK in the preserved formulation. Betaxolol was twice as effective at preventing the effect of IgG (34% to 37%) than that of zymosan (14%), regardless of the presence of BAK. However, BAK itself strongly aggravated complement activation by both activators. Carteolol, timolol, betaxolol and latanoprost did not activate complement system. On the contrary, the beta-blockers timolol and betaxolol exerted an anti-inflammatory effect by preventing complement activation. The deleterious effect of benzalkonium seems to have been neutralized within the preserved eyedrops through a mechanism that remains to be elucidated. Our study suggests that inflammatory signs in glaucoma patients should not be attributed to complement activation by antiglaucoma drugs.


[A study on enoxacin concentration in human prostatic tissue].
To examine the penetration of enoxacin into prostatic tissue of patients with benign prostatic hypertropHy, the serum concentration and prostatic tissue level of enoxacin were measured. Enoxacin was administered orally at a dose of 200 mg, three times 2 days and one day before and once on the day of the operation. Blood and prostatic tissue samples were taken during the operation. The mean concentration of enoxacin was 2.84 +/- 0.349 g/ml in serum and 4.60 +/- 0.631 micrograms/ml in the prostatic tissue.


In situ examination of the time-course for secondary mineralization of Haversian bone using synchrotron Fourier transform infrared microspectroscopy.
At the tissue level it is well established that the rate of remodeling is related to the degree of mineralization. However, it is unknown how long it takes for an individual bone structural unit (BSU) to become fully mineralized during secondary mineralization. Using synchrotron Fourier transform infrared microspectroscopy (FTIRM) we examined the time required for newly formed bone matrix to reach a physiological mineralization limit. Twenty-six, four-month old female New Zealand white rabbits were administered up to four different fluorochrome labels at specific time points to evaluate the chemical composition of labeled osteons from the tibial diaphysis that had mineralized for 1, 8, 18, 35, 70, 105, 140, 175, 210, 245, 280, 315, 350, and 385 days. Interstitial bone from 505 day old rabbits was used as a reference value for the physiological limit to which bone mineralizes. Using synchrotron FTIRM, area integrations were carried out on protein (Amide I: 1688-1623 cm(-1)), carbonate (v(2)CO(3)(2-): 905-825 cm(-1)), and phosphate (v(4)PO(4)(3-): 650-500 cm(-1)) IR bands. IR spectral data are presented as ratios of phosphate/protein (overall matrix mineralization) and carbonate/protein. The rate of mineralization of osteonal bone proceeded rapidly between day 1 and 18, reaching 67% of interstitial bone levels. This was followed by a slower, more progressive accumulation of mineral up to day 350. By 350 days the rate of increase plateaued. The ratio of carbonate/protein also increased rapidly during the first 18 days, reaching 73% of interstitial bone levels. The ratio of carbonate/protein plateaued by day 315, reaching levels not significantly different to interstitial bone levels. In conclusion, our data demonstrate that bone accumulates mineral rapidly during the first 18 days (primary mineralization), followed by a more gradual increase in the accumulation of mineral (secondary mineralization) which we found to be completed in 350 days.


Control of uncoupling protein in brown-fat mitochondria by purine nucleotides. Chemical modification by diazobenzenesulfonate.
The uncoupling protein (UP) of isolated brown adipose tissue mitochondria was studied with respect to the mechanism of control of UP function by purine nucleotides. Passive transport of H+ and Cl- was followed simultaneously in a KCl medium. With both GDP and ATP a higher sensitivity of Cl- transport (apparent Ki = 2.2 microM and 4.7 microM respectively) than of H+ transport (apparent Ki = 7.7 microM and 34 microM respectively) was observed. Chemical modification of isolated mitochondria by diazobenzenesulfonate (DABS) up to 75 mumol/mg protein did not affect the transport, its ionic selectivity and regulation by endogenous free fatty acids. In contrast, the sensitivity to purine nucleotides of both H+ and Cl- translocation was decreased (apparent Ki increased 71 and 47 times respectively). DABS decreased the affinity of [3H]GDP for the specific nucleotide-binding site on mitochondria (Kd increased from 2.7 microM to 13 microM) and depressed, to a smaller extent, the GDP-binding capacity. Correlation between occupancy of the specific nucleotide-binding site by GDP and inhibition of transport yielded a linear relationship for Cl- transport in control mitochondria. For H+ transport in the control, and for both H+ and Cl- transports in DABS-treated mitochondria, a biphasic correlation was obtained. The results show that different structural parts of UP are involved in transport and its control by the regulatory ligands and that, in addition to binding of purine nucleotides to UP, the inhibition of ion transport by purine nucleotides depends on an intrinsic factor modulating the inhibitory effect.


[Internship in a community hospital. Experience gained in the "Faculdade de Ciências Médicas e Biológicas de Botucatu" (author's transl)].
Currently the verification of the equivalence between an University Hospital (U.H.) and a Community Hospital (C.H.), in means of internship accomplishment is an important subject. The F.C.M.B.B. has accumulated experience in this matter, since among the 514 doctors graduated at this school, in the period of 1968 to 1973: 309 accomplished their internship at Botucatu (U.H.); and the remainers 205 at C.H.s. The author's purpose was to try to evaluate the existence, or not, of this equivalence. Thus two different questionnaries were elaborated, one that was answered by two kinds of intern's instructors (total 58): the teaching staff of U.H. (33), and the physicians belonging to the staff of one of the C.H.s. The other was submitted to two groups of ex-interns (total: 73): ex-interns from U.H. (53) and from C.H.s. (20). Both, ex-interns and teachers, were selected by chance. The answers were analysed by stablishing a comparison between the two groups: U.H. and C.H. The results were analysed by chi2 (chi-square) using the Yates correction for 2 X 2 tables. They seem to suggest that there is an equivalence between U.H.s. and C.H.s. in means of internship accomplishment, since certain characteristics are observed by the C.H.s.


Applications of Digital PCR for Clinical Microbiology.
Digital PCR (dPCR) is an important new tool for use in the clinical microbiology laboratory. Its advantages over quantitative PCR (qPCR), including absolute quantification without a standard curve, improved precision, improved accuracy in the presence of inhibitors, and more accurate quantitation when amplification efficiency is low, make dPCR the assay of choice for several specimen testing applications. This minireview will discuss the advantages and disadvantages of dPCR compared to qPCR, its applications in clinical microbiology, and considerations for implementation of the method in a clinical laboratory.


Vegetational association of host-seeking adult blacklegged ticks, Ixodes scapularis Say (Acari: Ixodidae), on dairy farms in northwestern Wisconsin.
As a measure of the risk for exposure to Lyme disease, we estimated the distribution of host-seeking adults of the blacklegged tick, Ixodes scapularis Say, on dairy farms in Barron County in northwestern Wisconsin. Vegetation ecotypes that were common to 18 farms that were representative of the county were surveyed by flag sampling. Tick prevalence and abundance, which were similar during fall and spring periods, were very low in farmhouse yards and forage croplands; only a single male was collected from 17 lawns. Sampling of 18 pastures with lactating cows also yielded only a single I. scapularis. In contrast, I. scapularis adults were captured in 9 of 37 samples from 18 pastures with heifers and dry cows; in those pastures, ticks were associated with woods and secondary vegetation or margins of pastures adjacent to woodlands. Blacklegged ticks were most prevalent and numerous in ungrazed woodlands; adults were captured in 27 of 53 samples on 13 of 15 farms, particularly when evidence of white-tailed deer, Odocoileus virginianus (Zimmermann), was apparent. The risk of encountering adult blacklegged ticks on dairy farms in Barron County, Wisconsin is therefore greatest in woodlands habitat. The presence of adult ticks in pastures with heifers and dry cows establishes an ecological basis for the exposure of dairy cattle to adult I. scapularis. Pastures with lactating cows, farmhouse yards, and forage croplands represent negligible risk.


A study on the tapping point drawing method for registration of maxillomandibular relationships in edentulous patients: Part I. Influence of the tapping force on the tapping point.
To investigate the registration of maxillomandibular relationships by the tapping point drawing method, the characteristics of tapping points were measured in ten edentulous subjects by tracing the tapping points when various tapping forces were used. In seven subjects, when tapping involved a heavy force, the tapping point position markedly shifted from the position when light force was used. The position of the tapping point shifted in the antero-posterior direction in five subjects and in the right-left direction in two others. The tapping point position of the other three subjects did not change when stronger tapping forces were applied; these tapping points either coincided with the apex of the gothic arch or included it. Furthermore, in all subjects, the tapping point size increased when the degree of tapping force increased. However, the degree of increase in tapping point size was smaller in the subjects in whom the tapping point position coincided with the apex of the gothic arch or included it. From these findings, we have confirmed that accurate tapping point tracing necessitates controlling the tapping force.


Functional characterization of a member of alanine or glycine: cation symporter family in halotolerant cyanobacterium Aphanothece halophytica.
Membrane proteins of amino acid-polyamine-organocation (APC) superfamily transport amino acids and amines across membranes and play important roles in the regulation of cellular processes. The alanine or glycine: cation symporter (AGCS) family belongs to APC superfamily and is found in prokaryotes, but its substrate specificity remains to be clarified. In this study, we found that a halotolerant cyanobacterium, Aphanothece halophytica has two putative ApagcS genes. The deduced amino acid sequence of one of genes, ApagcS1, exhibited high homology to Pseudomonas AgcS. The ApagcS1 gene was expressed in Escherichia coli JW4166 which is deficient in glycine uptake. Kinetics studies in JW4166 revealed that ApAgcS1 is a sodium-dependent glycine transporter. Competition experiments showed the significant inhibition by glutamine, asparagine, and glycine. The level of mRNA for ApagcS1 was induced by NaCl and nitrogen-deficient stresses. Uptake of glutamine by ApAgcS1 was also observed. Based on these data, the physiological role of ApAgcS1 was discussed.


Overcoming the organization-practice barrier in sports injury prevention: A nonhierarchical organizational model.
The organization of sports at the national level has seldom been included in scientific discussions of sports injury prevention. The aim of this study was to develop a model for organization of sports that supports prevention of overuse injuries. The quality function deployment technique was applied in seminars over a two-season period to develop a national organizational structure for athletics in Sweden that facilitates prevention of overuse injuries. Three central features of the resulting model for organization of sports at the national level are (a) diminishment of the organizational hierarchy: participatory safety policy design is introduced through annual meetings where actors from different sectors of the sporting community discuss training, injury prevention, and sports safety policy; (b) introduction of a safety surveillance system: a ubiquitous system for routine collection of injury and illness data; and (c) an open forum for discussion of safety issues: maintenance of a safety forum for participants from different sectors of the sport. A nonhierarchical model for organization of sports at the national level - facilitated by modern information technology - adapted for the prevention of overuse injuries has been developed. Further research is warranted to evaluate the new organizational model in prospective effectiveness studies.


Coping skills and mental health status in adolescents when a parent has cancer: a multicenter and multi-perspective study.
Parental cancer increases the risk of psychosocial problems in adolescents. We investigated the frequency and efficacy of adolescents' coping strategies and relationships between those strategies and mental health status. Age and gender differences regarding coping and mental health were also investigated. In total, 214 adolescents from 167 families participated in a cross-sectional, multicenter study. All participants were recruited from standard oncological care. Among the participants, 52% utilized a child-centered intervention program. Adolescents' coping skills were measured using KIDCOPE. Mental health status was rated by adolescents and parents by the SDQ for symptomatology and the KIDSCREEN for well-being. We found that 29% of the adolescents showed emotional and behavioral problems. We found gender differences in mental health status but not in coping. Adolescents used a broad spectrum of coping strategies. Active problem-solving, distraction, acceptance, wishful thinking and seeking social support were the most frequently used coping strategies. The utilization of certain coping skills was mediated by their perceived efficacy. Problem-focused or approach-oriented coping strategies generally are associated with better mental health, while avoidance-oriented coping are associated with worse mental health. Emotion-focused coping was associated with both lower and higher mental health. The strategies used by adolescents to cope with parental cancer are associated with their mental health. Problem-solving and approach-oriented coping strategies should be facilitated by psychological interventions regardless of age and gender. Age and gender differences in adolescents' mental health should be further investigated because these differences are not explained by differences in coping strategies.


Aortic Arch Debranching and Thoracic Endovascular Aortic Repair (TEVAR) for Type B Aortic Dissection.
Since its introduction, thoracic endovascular aortic repair (TEVAR) has revolutionized the treatment of type B aortic dissections (TBADs). However, the proximal aspect of the aortic pathology treated may infringe on the origin of the left subclavian artery or even more proximally. Hence, to ensure durable outcomes, the origin of these vessels needs to be covered, but an extra-anatomical bypass is required to perfuse vital branches, known as aortic arch debranching. This series aims to describe and delineate the disparities of aortic arch debranching during TEVAR for TBAD. A retrospective review and analysis of a multicenter international database was conducted to identify patients with TBAD treated with TEVAR between 2005 and 2021. Data analyzed included patient demographics, disease characteristics, operative characteristics, and postoperative outcomes with follow-up on mortality and reintervention. All statistical analyses were carried out using IBM SPSS 26. Patient survival was calculated using a Kaplan-Meier survival analysis, and a P value of less than 0.05 was considered statistically significant. A total of 58 patients were included in the analysis, of which 27 (46.6%) presented with complicated disease and 31 were uncomplicated, of which 10 (17.2%) were classed as high risk and 21 (36.2%) low risk. Zone 2 was the most common proximal landing zone for the stent graft. Left subclavian artery bypass was performed selectively (26%), with 1 stroke occurring, likely due to embolic reasons. A further 6 underwent more proximal aortic debranching before TEVAR (10%) and was a significant risk factor for mortality and the number of stents deployed. The overall rates of reintervention and mortality were 17.2% (n = 10) and 29.3% (n = 17). Aortic arch debranching and TEVAR for TBAD is associated with significant mortality. Future developments to treat aortic arch pathology could incorporate branched graft devices, eliminating the need for debranching, improving stroke rates, and reducing future reinterventions.


The Northwestern University Twin Study. VII: The mode of delivery in twin pregnancy. North American considerations.
No consensus exists for the optimal mode of delivery for twin fetuses. Opinions vary by type of institution (university medical center vs community hospital), country or continent (North America vs Western Europe) and personal preference of individual physicians. This article lists clinical considerations in arriving at the decision and presents them in the form of a decision tree.


Glucocorticoids stimulate plasminogen activator production by rat granulosa cells.
We studied the direct effects of glucocorticoids on plasminogen activator (PA) production by rat granulosa cells. PA production was assayed by culturing rat granulosa cells on [125I]fibrin plates and determining the extent of fibrinolysis after addition of the specific substrate plasminogen. In granulosa cells from preantral follicles of immature rats, treatment with FSH caused dose-dependent increases in PA production whereas glucocorticoids by itself was without effect. Increasing concentrations (10(-10) to 10(-6) M) of both natural and synthetic glucocorticoids potentiated the stimulating effect of FSH on PA production by 120 to 170%. The stimulatory potencies of the natural corticosteroids correlated with the glucocorticoid potencies (cortisol/corticosterone greater than aldosterone/11-deoxycorticosterone). In granulosa cells from Graafian follicles of mature rats, glucocorticoids on its own had direct stimulating effect on PA production. The stimulatory action of glucocorticoids on FSH-dependent PA production was completely blocked by simultaneous treatment with antiglucocorticoid RU 486. Antiserum directed against tissue-type PA (tPA) neutralized the increased fibrinolytic actions of glucocorticoids. Using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fibrin autography techniques, we showed that the increase in fibrinolytic activity in response to glucocorticoids resulted from increased production of tPA rather than urokinase-like PA. The effect of glucocorticoids on the production of PA inhibitors (PAI) was examined by 1) neutralization of urokinase activity by increasing amounts of culture media from granulosa cells treated with glucocorticoids, 2) reverse fibrin autography, and 3) Western blot analysis with a specific PAI antiserum. All these methods failed to detect a stimulatory action of glucocorticoids, with or without FSH, on PAI production by rat granulosa cells in the culture media. Our data showed that glucocorticoids have a direct stimulating effect on tPA production, but unlike its action on other in vitro systems, they have no significant effect on PAI production by rat granulosa cells in vitro.


Comparative forelimb morphology of scratch-digging and chisel-tooth digging African mole-rat species.
Bathyergus suillus (Cape dune mole-rat) and Heterocephalus glaber (naked mole-rat) are two species of subterranean burrowing rodents. Bathyergus suillus occurs in soft sandy soils and is regarded as a scratch-digger, while H. glaber is found in hard, compact soils and is a chisel-tooth digging species. The present study aimed to determine musculoskeletal differences in the forelimb of these two species. The muscles of the forelimb, back and neck were dissected to the points of origin and insertion in the left and right forelimbs, B. suillus (n = 7) and H. glaber (n = 5). Dissected muscles were photographed before maceration to demonstrate muscle attachments. The scapular spine, acromion process and clavicle were relatively straight in B. suillus. In comparison a curved scapular spine, acromion process and clavicle were observed in H. glaber. In both species, the clavicle rested on the greater tuberosity of the humerus. In B. suillus, the deltoid tuberosity was prominent and situated more distally on the humeral shaft compared to the indistinct, more proximally situated deltoid tuberosity in H. glaber. A prominent bony structure underlying the thenar pad as well as a cartilaginous protrusion beneath the hypothenar pad were observed on the palmar surface of the manus in B. suillus. Prominent claws were observed in B. suillus. A robust m. sternohyoideus was observed in H. glaber while mm. tensor fasciae antebrachii and coracobrachialis were absent. The flexors of the antebrachium of B. suillus had additional and enlarged attachment sites. The forelimb of B. suillus may be morphologically adapted for scratch-digging with relatively large and additional forelimb muscles and robust bones. In comparison, H. glaber had a reduction in the relative size, amount of muscles as well as number of attachment sites in the forelimb muscles, while the well-developed ventral neck muscles may facilitate neck and head stabilisation during chisel-tooth digging.


Mycotoxins and beer. Impact of beer production process on mycotoxin contamination. A review.
Beer is the most consumed alcoholic beverage in the world. Its contamination with mycotoxins is of public health concern, especially for heavy drinkers. Beer production implies a variety of operations which might impact the initial level of mycotoxins in a positive or negative way. The complexity of these operations do not give to the brewer a complete control on chemical and biochemical reactions that take place in the batch, but the knowledge about mycotoxin properties can help in identifying the operations decreasing their level in foodstuffs and in the development of mitigation strategies. This review discusses available data about mycotoxin evolution during malting and brewing process. The operations that may lead to a decrease in mycotoxin load are found to be steeping, kilning, roasting, fermentation and stabilization operations applied over the process (e.g. clarification). Also, other general decontamination strategies usually employed in food industry, such as hot water treatment of barley, ozonation or even the use of lactic acid bacteria starter cultures during malting or fermentation are considered.


Cardiac sarcoidosis - an expert review for the chest physician.
Introduction: Sarcoidosis is a multisystem granulomatous disease predominantly affecting the lungs, with increased risk of cardiovascular disease, pulmonary hypertension and cardiac sarcoidosis (CS), the latter due to direct granuloma infiltration. Sarcoidosis is often managed by chest physicians who need to understand the diagnostic pathways and initial management plans for patients with cardiac involvement. Areas covered: The most serious consequence of CS is sudden cardiac death due to ventricular tachyarrhythmias or complete atrioventricular block. Additional complications include atrial arrhythmias and congestive cardiac failure. There are no internationally accepted screening pathways, but a combination of history, clinical examination and ECG detects up to 85% of cases. Newer modalities including signal-averaged ECG and speckle-tracking echocardiography increase identification of patients who require a definitive diagnosis. Early immunosuppression reduces the risk of conduction abnormalities and incidence of supraventricular arrhythmias. Management of ventricular arrhythmias requires antiarrhythmic medications followed by possible catheter ablation and device (ICD) implantation. Expert commentary: Prospective trials are underway to identify the optimum methods for screening, which will guide future international statements on indications for and methods of screening in CS.


Physiology and pathophysiology of the DUOXes.
The dual oxidases (DUOXes) 1 and 2 are named based on their having both a domain homologous to the NADPH-oxidase of the phagocyte NADPH-oxidase gp91( phox )/NOX2 and a domain homologous to thyroid peroxidase. The DUOX1 and DUOX2 mRNAs were originally cloned from thyroid tissue, and the corresponding proteins were recognized as intricate components of the thyroid hormone synthesis process, providing hydrogen peroxide essential for the organification of iodide. The function of DUOX2 in thyroid hormonogenesis has been firmly established by linking the congenital hypothyroid phenotype "total iodide organification defect" to biallelic inactivating DUOX2 mutations. Based on the expression of both DUOXes in combination with a peroxidase in a range of different tissues and functional studies; the concept evolves that DUOX is important not only for thyroid hormonogenesis but also as an integral part of the host defense system of mucosal surfaces, participates in the control of epithelial infection, augments surface B-cell receptor signaling in lymphocytes, and is involved in generating a respiratory burst at fertilization.


Time-resolved single-photon detection module based on silicon photomultiplier: A novel building block for time-correlated measurement systems.
We present the design and preliminary characterization of the first detection module based on Silicon Photomultiplier (SiPM) tailored for single-photon timing applications. The aim of this work is to demonstrate, thanks to the design of a suitable module, the possibility to easily exploit SiPM in many applications as an interesting detector featuring large active area, similarly to photomultipliers tubes, but keeping the advantages of solid state detectors (high quantum efficiency, low cost, compactness, robustness, low bias voltage, and insensitiveness to magnetic field). The module integrates a cooled SiPM with a total photosensitive area of 1 mm(2) together with the suitable avalanche signal read-out circuit, the signal conditioning, the biasing electronics, and a Peltier cooler driver for thermal stabilization. It is able to extract the single-photon timing information with resolution better than 100 ps full-width at half maximum. We verified the effective stabilization in response to external thermal perturbations, thus proving the complete insensitivity of the module to environment temperature variations, which represents a fundamental parameter to profitably use the instrument for real-field applications. We also characterized the single-photon timing resolution, the background noise due to both primary dark count generation and afterpulsing, the single-photon detection efficiency, and the instrument response function shape. The proposed module can become a reliable and cost-effective building block for time-correlated single-photon counting instruments in applications requiring high collection capability of isotropic light and detection efficiency (e.g., fluorescence decay measurements or time-domain diffuse optics systems).


Compartmentalization of SHC, GRB2 and mSOS, and hyperphosphorylation of Raf-1 by EGF but not insulin in liver parenchyma.
Rat liver parenchyma harbors equal numbers of epidermal growth factor (EGF) and insulin receptors. Following administration of a saturating dose of EGF (10 micrograms/100 g body weight), there was a rapid (t1/2 approximately 1.1 min) internalization of receptor coincident with its tyrosine phosphorylation at residue 1173 and receptor recruitment of the adaptor protein SHC, its tyrosine phosphorylation and its association with GRB2 and the Ras guanine nucleotide exchange factor, mSOS, largely in endosomes. This led to a cytosolic pool of a complex of tyrosine-phosphorylated SHC, GRB2 and mSOS. It was demonstrated that these constituents were linked to Ras activation by the characteristic decrease in Raf-1 mobility on SDS-PAGE, which was maintained for 60 min after a single bolus of administered EGF. While insulin administration (15 micrograms/100 g body weight) led to insulin receptor beta-subunit tyrosine phosphorylation and internalization, there was little detectable tyrosine phosphorylation of SHC, recruitment of GRB2, association of a complex with mSOS or any detectable change in the mobility of Raf-1. Therefore, in normal physiological target cells in vivo, distinct signaling pathways are realized after EGF or insulin receptor activation, with regulation of this specificity most probably occurring at the locus of the endosome.


Oligomeric α-synuclein is increased in basal tears of Parkinson's patients.
Aim: Secretion of proteins into basal tears of Parkinson's disease (PD) patients may be altered by changes in nerve function. Materials & methods: Oligomeric α-SynOligo and total α-SynTotal, CCL-2, DJ-1, LF and MMP-9 were measured in basal tears from 93 PD patients and 82 age- and sex-equivalent healthy controls. Results: α-SynTotal was decreased (p = 0.0043), whereas α-SynOligo (p < 0.0001) and the ratio of α-SynOligo/α-SynTotal (p < 0.0001) were increased in basal tears from PD patients compared with healthy controls. Area under receiver-operating curves of α-SynOligo and α-SynOligo/α-SynTotal contents were 0.70 (95% confidence limits: 0.621-0.774) and 0.72 (95% confidence limits: 0.642-0.792). Conclusion: PD patient basal tears may contain biomarkers that can be assayed noninvasively and inexpensively.


Myofibroblasts acquire retinoic acid-producing ability during fibroblast-to-myofibroblast transition following kidney injury.
Tubular injury and interstitial fibrosis are the hallmarks of chronic kidney disease. While recent studies have verified that proximal tubular injury triggers interstitial fibrosis, the impact of fibrosis on tubular injury and regeneration remains poorly understood. We generated a novel mouse model expressing diphtheria toxin receptor on renal fibroblasts to allow for the selective disruption of renal fibroblast function. Administration of diphtheria toxin induced upregulation of the tubular injury marker Ngal and caused tubular proliferation in healthy kidneys, whereas administration of diphtheria toxin attenuated tubular regeneration in fibrotic kidneys. Microarray analysis revealed down-regulation of the retinol biosynthesis pathway in diphtheria toxin-treated kidneys. Healthy proximal tubules expressed retinaldehyde dehydrogenase 2 (RALDH2), a rate-limiting enzyme in retinoic acid biosynthesis. After injury, proximal tubules lost RALDH2 expression, whereas renal fibroblasts acquired strong expression of RALDH2 during the transition to myofibroblasts in several models of kidney injury. The retinoic acid receptor (RAR) RARγ was expressed in proximal tubules both with and without injury, and αB-crystallin, the product of an RAR target gene, was strongly expressed in proximal tubules after injury. Furthermore, BMS493, an inverse agonist of RARs, significantly attenuated tubular proliferation in vitro. In human biopsy tissue from patients with IgA nephropathy, detection of RALDH2 in the interstitium correlated with older age and lower kidney function. These results suggest a role of retinoic acid signaling and cross-talk between fibroblasts and tubular epithelial cells during tubular injury and regeneration, and may suggest a beneficial effect of fibrosis in the early response to injury.


Function of arginine-166 in the active site of Escherichia coli alkaline phosphatase.
The function of arginine residue 166 in the active site of Escherichia coli alkaline phosphatase was investigated by site-directed mutagenesis. Two mutant versions of alkaline phosphatase, with either serine or alanine in the place of arginine at position 166, were generated by using a specially constructed M13 phage carrying the wild-type phoA gene. The mutant enzymes with serine and alanine at position 166 have very similar kinetic properties. Under conditions of no external phosphate acceptor, the kcat for the mutant enzymes decreases by approximately 30-fold while the Km increases by less than 2-fold. When kinetic measurements are carried out in the presence of a phosphate acceptor, 1.0 M Tris, the kcat for the mutant enzymes is reduced by less than 3-fold, while the Km increases by more than 50-fold. For both mutant enzymes, in either the absence or the presence of a phosphate acceptor, the catalytic efficiency as measured by the kcat/Km ratio decreases by approximately 50-fold as compared to the wild type. Measurements of the Ki for inorganic phosphate show an increase of approximately 50-fold for both mutants. Phenylglyoxal, which inactivates the wild-type enzyme, does not inactivate the Arg-166----Ala enzyme. This result indicates that Arg-166 is the same arginine residue that when chemically modified causes loss of activity [Daemen, F.J.M., & Riordan, J.F. (1974) Biochemistry 13, 2865-2871]. The data reported here suggest that although Arg-166 is important for activity is not essential. The analysis of the kinetic data also suggests that the loss of arginine-166 at the active site of alkaline phosphatase has two different effects on the enzyme. First, the binding of the substrate, and phosphate as a competitive inhibitor, is reduced; second, the rate of hydrolysis of the covalent phosphoenzyme may be diminished.


Profiling Temporal Changes of the Pineal Transcriptomes at Single Cell Level Upon Neonatal HIBD.
Neonatal hypoxic-ischemic brain damage (HIBD) often results in various neurological deficits. Among them, a common, yet often neglected, symptom is circadian rhythm disorders. Previous studies revealed that the occurrence of cysts in the pineal gland, an organ known to regulate circadian rhythm, is associated with circadian problems in children with HIBD. However, the underlying mechanisms of pineal dependent dysfunctions post HIBD remain largely elusive. Here, by performing 10x single cell RNA sequencing, we firstly molecularly identified distinct pineal cell types and explored their transcriptome changes at single cell level at 24 and 72 h post neonatal HIBD. Bioinformatic analysis of cell prioritization showed that both subtypes of pinealocytes, the predominant component of the pineal gland, were mostly affected. We then went further to investigate how distinct pineal cell types responded to neonatal HIBD. Within pinealocytes, we revealed a molecularly defined β to α subtype conversion induced by neonatal HIBD. Within astrocytes, we discovered that all three subtypes responded to neonatal HIBD, with differential expression of reactive astrocytes markers. Two subtypes of microglia cells were both activated by HIBD, marked by up-regulation of Ccl3. Notably, microglia cells showed substantial reduction at 72 h post HIBD. Further investigation revealed that pyroptosis preferentially occurred in pineal microglia through NLRP3-Caspase-1-GSDMD signaling pathway. Taken together, our results delineated temporal changes of molecular and cellular events occurring in the pineal gland following neonatal HIBD. By revealing pyroptosis in the pineal gland, our study also provided potential therapeutic targets for preventing extravasation of pineal pathology and thus improving circadian rhythm dysfunction in neonates with HIBD.


Interferon-free treatment for patients with chronic hepatitis C and autoimmune liver disease: higher SVR rates with special precautions for deterioration of autoimmune hepatitis.
Interferon-free treatment can achieve higher sustained virological response (SVR) rates, even in patients in whom hepatitis C virus (HCV) could not be eradicated in the interferon treatment era. Immune restoration in the liver is occasionally associated with HCV infection. We examined the safety and effects of interferon-free regimens on HCV patients with autoimmune liver diseases. All 7 HCV patients with autoimmune hepatitis (AIH) completed treatment and achieved SVR. Three patients took prednisolone (PSL) at baseline, and 3 did not take PSL during interferon-free treatment. In one HCV patient with AIH and cirrhosis, PSL were not administered at baseline, but she needed to take 40 mg/day PSL at week 8 for liver dysfunction. She also complained back pain and was diagnosed with vasospastic angina by coronary angiography at week 11. However, she completed interferon-free treatment. All 5 HCV patients with primary biliary cholangitis (PBC) completed treatment and achieved SVR. Three of these HCV patients with PBC were treated with UDCA during interferon-free treatment. Interferon-free regimens could result in higher SVR rates in HCV patients with autoimmune liver diseases. As interferon-free treatment for HCV may have an effect on hepatic immunity and activity of the autoimmune liver diseases, careful attention should be paid to unexpected adverse events in their treatments. Total 12 patients with HCV and autoimmune liver diseases [7 AIH and PBC], who were treated with interferon-free regimens, were retrospectively analyzed.


The increasing relevance of immunobiology within a connected animal science curriculum.
Modern technological agriculture emerged in the 20th century and has expanded into a global enterprise occupying approximately 38% of the Earth's land area and accounting for over 40% of the world's workforce. The United Nations Food and Agriculture Organization estimates that to feed a world population of 9-billion people in 2050 will require an almost doubling of overall food production, including meat, dairy, and egg production over 2010 levels. However, our collective ability to meet this demand cannot be taken for granted. Despite many successes, global agricultural systems now face multiple unprecedented challenges including a dearth of new treatments for livestock diseases. The discovery of antibiotics led to a complacency now reflected in a dependency on exogenous antimicrobials and a growing threat of antimicrobial resistance. Developments within the field of immunobiology had led to significant breakthroughs in understanding of human health and disease. However, despite over 60% of infectious diseases being zoonotic in nature and nonhuman animals acting as an important disease reservoir, research in livestock immunobiology has not been as resourced. As a direct result, recalcitrant animal diseases continue to threaten sustainability of animal production systems, security of the food chain and human health. It is within the context of collective One Health action that ambitious innovation in the connectivity of animal science undergraduate curricula is urgently required, specifically to include threshold concepts in immunobiology. Fostering transformative learning is critical to equip future generations of animal scientists with the knowledge and interdisciplinary skills to counter these existential challenges of our time.


Comparison of hospitalization outcomes for delivery and resource utilization between pregnant women with kidney transplants and chronic kidney disease in the United States.
This study aimed to assess outcomes of delivery hospitalizations, including acute kidney injury (AKI), obstetric and foetal events and resource utilization among pregnant women with kidney transplants compared with pregnant women with no known kidney disease and those with chronic kidney disease (CKD) Stages 3-5. Hospitalizations for delivery in the US were identified using the enhanced delivery identification method in the National Inpatient Sample dataset from the years 2009 to 2014. Diagnoses of CKD Stages 3-5, kidney transplantation, along with obstetric events, delivery methods and foetal events were identified using ICD-9-CM diagnosis and procedure codes. Patients with no known kidney disease group were identified by excluding any diagnoses of CKD, end stage kidney disease, and kidney transplant. Multivariable logistic regression accounting for the survey weights and matched regression was conducted to investigate the risk of maternal and foetal complications in women with kidney transplants, compared with women with no kidney transplants and no known kidney disease, and to women with CKD Stages 3-5. A total of 5, 408, 215 hospitalizations resulting in deliveries were identified from 2009 to 2014, including 405 women with CKD Stages 3-5, 295 women with functioning kidney transplants, and 5, 405, 499 women with no known kidney disease. Compared with pregnant women with no known kidney disease, pregnant kidney transplant recipients were at higher odds of hypertensive disorders of pregnancy (OR = 3.11, 95% CI [2.26, 4.28]), preeclampsia/eclampsia/HELLP syndrome (OR = 3.42, 95% CI [2.54, 4.60]), preterm delivery (OR = 2.46, 95% CI [1.75, 3.45]), foetal growth restriction (OR = 1.74, 95% CI [1.01, 3.00]) and AKI (OR = 10.46, 95% CI [5.33, 20.56]). There were no significant differences in rates of gestational diabetes or caesarean section. Pregnant women with kidney transplants had 1.30-times longer lengths of stay and 1.28-times higher costs of hospitalization. However, pregnant women with CKD Stages 3-5 were at higher odds of AKI (OR = 5.29, 95% CI [2.41, 11.59]), preeclampsia/eclampsia/HELLP syndrome (OR = 1.72, 95% CI [1.07, 2.76]) and foetal deaths (OR = 3.20, 95% CI [1.06, 10.24]), and had 1.28-times longer hospital stays and 1.37-times higher costs of hospitalization compared with pregnant women with kidney transplant. Pregnant women with kidney transplant were more likely to experience adverse events during delivery and had longer lengths of stay and higher total charges when compared with women with no known kidney disease. However, pregnant women with moderate to severe CKD were more likely to experience serious complications than kidney transplant recipients.


Beating heart minimally invasive mitral valve surgery in patients with previous sternotomy: the operative technique and early outcomes.
Reoperative mitral valve surgery is increasingly required and can be associated with significant morbidity and mortality. The beating heart minimally invasive mitral valve surgery has a proposed benefit in avoiding the risks of repeat sternotomy, with reducing the need for adhesiolysis and cardioplegia reperfusion injury. We describe our experience with such a technique in patients with previous sternotomy. A retrospective study was performed and all patients undergoing surgery of mitral valve through a right limited thoracotomy without application of an aortic cross-clamp (beating heart) as a redo cardiac surgery between January 2006 and January 2015 were included (n=25). Perioperative data as well as the operative technique are presented. Six patients (24%) had two previous sternotomies and one (4%) had three previous sternotomies. Mitral valve repair was performed in 11 patients (44%). No patient required conversion to median sternotomy. Inotropic support beyond 4 hours after operation was required in seven patients (28%). Ventilation time was less than 12 hours in 14 patients (56%) with another six patients (24%) extubated within 24 hours after surgery. Postoperative course was complicated with cerebrovascular accident in two patients (8%). In-hospital mortality was 4% (n=1). There was no 30-day mortality after discharge. Reoperative mitral valve surgery can be safely performed through a limited right thoracotomy approach on a beating heart while on full cardiopulmonary bypass. The technique can be associated with potentially shorter operation, shorter cardiopulmonary bypass and a less complicated recovery.


Global and regional child deaths due to injuries: an assessment of the evidence.
Injuries result in substantial number of deaths among children globally. The burden across many settings is largely unknown. We estimated global and regional child deaths due to injuries from publicly available evidence. We searched for community-based studies and nationally representative data reporting on child injury deaths published after year 1990 from CINAHL, EMBASE, IndMed, LILACS, Global Health, MEDLINE, SCOPUS, and Web of Science. Specific and all-cause mortality due to injuries were extracted for three age groups (0-11 months, 1-4 years, and 0-4 years). We conducted random-effects meta-analysis on extracted crude estimates, and developed a meta-regression model to determine the number of deaths due to injuries among children aged 0-4 years globally and across the World Health Organization (WHO) regions. Twenty-nine studies from 16 countries met the selection criteria. A total of 230 data-points on 15 causes of injury deaths were retrieved from all studies. Eighteen studies were rated as high quality, although heterogeneity was high (I2 = 99.7%, P < 0.001) reflecting variable data sources and study designs. For children aged 0-11 months, the pooled crude injury mortality rate was 29.6 (95% confidence interval (CI) = 21.1-38.1) per 100 000 child population, with asphyxiation being the leading cause of death (neonatal) at 189.1 (95% CI = 142.7-235.4) per 100 000 followed by suffocation (post-neonatal) at 18.7 (95% CI = 11.8-25.7) per 100 000. Among children aged 1-4 years, the pooled crude injury mortality rate was 32.7 (95% CI = 27.3-38.1) per 100 000, with traffic injuries and drowning the leading causes of deaths at 10.8 (95% CI = 8.9-12.8) and 8.8 (95% CI = 7.5-10.2) per 100 000, respectively. Among children under five years, the pooled injury mortality rate was 37.7 (95% CI = 32.7-42.7) per 100 000, with traffic injuries and drowning also the leading causes of deaths at 10.3 (95% CI = 8.8-11.8) and 8.9 (95% CI = 7.8-9.9) per 100 000 respectively. When crude mortality changes over age, WHO regions, and study period were accounted for in our model, we estimated that in 2015 there were 522 167 (95% CI = 395 823-648 630) deaths among children aged 0-4 years, with South East Asia (SEARO) recording the highest number of deaths at 195 084 (95% CI = 159476-230502), closely followed by the Africa region (AFRO) with 176523 (95% CI = 115 040-237 831) deaths. Globally, traffic injuries and drowning were the leading causes of under-five injury fatalities in 2015 with 142 661 (22.0/100 000) and 123 270 (19.0/100 000) child deaths, respectively. The exception being burns in AFRO with 57 784 deaths (38.6/100 000). Varying study designs, case definitions, and particularly limited country representation from Africa and South-East Asia (where we reported higher estimates), imply a need for more studies for better population representative estimates. This study may have however provided improved understanding on child injury death profiles needed to guide further research, policy reforms and relevant strategies globally.


Metabolic activity, urease production, antibiotic resistance and virulence in dual species biofilms of Staphylococcus epidermidis and Staphylococcus aureus.
In this paper, the metabolic activity in single and dual species biofilms of Staphylococcus epidermidis and Staphylococcus aureus isolates was investigated. Our results demonstrated that there was less metabolic activity in dual species biofilms compared to S. aureus biofilms. However, this was not observed if S. aureus and S. epidermidis were obtained from the same sample. The largest effect on metabolic activity was observed in biofilms of S. aureus Mu50 and S. epidermidis ET-024. A transcriptomic analysis of these dual species biofilms showed that urease genes and genes encoding proteins involved in metabolism were downregulated in comparison to monospecies biofilms. These results were subsequently confirmed by phenotypic assays. As metabolic activity is related to acid production, the pH in dual species biofilms was slightly higher compared to S. aureus Mu50 biofilms. Our results showed that S. epidermidis ET-024 in dual species biofilms inhibits metabolic activity of S. aureus Mu50, leading to less acid production. As a consequence, less urease activity is required to compensate for low pH. Importantly, this effect was biofilm-specific. Also S. aureus Mu50 genes encoding virulence-associated proteins (Spa, SplF and Dps) were upregulated in dual species biofilms compared to monospecies biofilms and using Caenorhabditis elegans infection assays, we demonstrated that more nematodes survived when co-infected with S. epidermidis ET-024 and S. aureus mutants lacking functional spa, splF or dps genes, compared to nematodes infected with S. epidermidis ET-024 and wild- type S. aureus. Finally, S. epidermidis ET-024 genes encoding resistance to oxacillin, erythromycin and tobramycin were upregulated in dual species biofilms and increased resistance was subsequently confirmed. Our data indicate that both species in dual species biofilms of S. epidermidis and S. aureus influence each other's behavior, but additional studies are required necessary to elucidate the exact mechanism(s) involved.


Evaluation of a forage allocation model for Theodore Roosevelt National Park.
We developed a forage allocation model using a deterministic, linear optimization module in a commercially available spreadsheet package to help resource managers in Theodore Roosevelt National Park (TRNP), North Dakota determine optimum numbers of four ungulate species, bison (Bison bison), elk (Cervus elaphus), mule deer (Odocoileus hemionus), and feral horses, in the Park. TRNP staff actively managed bison, elk, and feral horse numbers within bounds suggested by our model from 1983 to 1996. During this period, we measured vegetation at 8 grassland and 12 wooded sites at 1-3 year intervals to determine if model solutions were appropriate for maintaining stable conditions in important plant communities in the Park. The data we recorded at these sites indicated minimal change in plant communities from 1983 to 1996. Changes in most vegetation categories that we expected when animal numbers exceeded model optimums for short periods (decreases in coverage/stem numbers of palatable plant species, increases in bare ground or unpalatable plant species) did not occur consistently under high or low precipitation conditions. The lack of sensitivity of our model to decreases in overall production of palatable plant species that occurred due to drought, fire, expansion of black-tailed prairie dog (Cynomys ludovicianus) colonies, and the spread of leafy spurge (Euphorbia esula) in areas of the Park where we did not have monitoring sites suggested that the model under-estimated the total number of ungulates that the Park could support. Management for population levels of ungulates defined by the model probably led to over protection of common plant communities and insufficient protection of rare plant communities. Detecting changes in rare plant communities could have been accomplished by re-designing our vegetation monitoring program, but changing emphasis to protection of rare plants would have likely promoted under use of grazing-tolerant habitat types, dissatisfaction in tourists visiting the Park to see large mammals, and large increases in cost and intrusiveness of management activities such as fencing and control of ungulate populations. The model was a flawed representation of grazing dynamics in TRNP, but we believe it succeeded in making management personnel aware of the biological constraints they face when making management decisions.


[Chemical constituents of ragweed (Ambrosia artemisiifolia L.)].
Three sesquiterpene lactones isolated from the aerial parts of Ambrosia artemisiifolia were identified as psilostachyin, psilostachyin B and psilostachyin C. All of them have some molluscicidal activity against Oncomelania hupensis.


Morphological and Optical Engineering for High-Performance Polymer Solar Cells.
We demonstrate morphological and optical engineering by using processing additives and optical spacers for polymer solar cells. Among various processing additives, introduction of diphenyl ether (DPE) into the active layer results in the smoothest surface roughness with uniform and well-distributed donor/acceptor domains, and the device with DPE shows the highest device efficiency of 10.22% due to enhanced charge collection efficiency and minimized recombination loss. Additional ZnO optical spacers on the active layer controls the distribution of the electric field in the whole device and enhances the light absorption within the active layer, thereby improving device efficiency up to 10.81%.


Structural origins of amino acid selection without editing by cysteinyl-tRNA synthetase.
Cysteinyl-tRNA synthetase (CysRS) is highly specific for synthesis of cysteinyl adenylate, yet does not possess the amino acid editing activity characteristic of many other tRNA synthetases. To elucidate how CysRS is able to distinguish cysteine from non-cognate amino acids, crystal structures of the Escherichia coli enzyme were determined in apo and cysteine-bound states. The structures reveal that the substrate cysteine thiolate forms a single direct interaction with a zinc ion bound at the base of the active site cleft, in a trigonal bipyramidal geometry together with four highly conserved protein side chains. Cysteine binding induces movement of the zinc ion towards substrate, as well as flipping of the conserved Trp205 indole ring to pack on the thiol side chain. The imidazole groups of five conserved histidines lie adjacent to the zinc ion, forming a unique arrangement suggestive of functional significance. Thus, amino acid discrimination without editing arises most directly from the favorable zinc-thiolate interaction, which is not possible for non-cognate substrates. Additional selectivity may be generated during the induced-fit conformational changes that help assemble the active site.


Substrate preference of Gen endonucleases highlights the importance of branched structures as DNA damage repair intermediates.
Human GEN1 and yeast Yen1 are endonucleases with the ability to cleave Holliday junctions (HJs), which are proposed intermediates in recombination. In vivo, GEN1 and Yen1 function secondarily to Mus81, which has weak activity on intact HJs. We show that the genetic relationship is reversed in Drosophila, with Gen mutants having more severe defects than mus81 mutants. In vitro, DmGen, like HsGEN1, efficiently cleaves HJs, 5΄ flaps, splayed arms, and replication fork structures. We find that the cleavage rates for 5΄ flaps are significantly higher than those for HJs for both DmGen and HsGEN1, even in vast excess of enzyme over substrate. Kinetic studies suggest that the difference in cleavage rates results from a slow, rate-limiting conformational change prior to HJ cleavage: formation of a productive dimer on the HJ. Despite the stark difference in vivo that Drosophila uses Gen over Mus81 and humans use MUS81 over GEN1, we find the in vitro activities of DmGen and HsGEN1 to be strikingly similar. These findings suggest that simpler branched structures may be more important substrates for Gen orthologs in vivo, and highlight the utility of using the Drosophila model system to further understand these enzymes.


Selective fluorescence quenching of 2,3-diazabicyclo[2.2.2]oct-2-ene by nucleotides.
[reaction: see text] The fluorescence quenching of 2,3-diazabicyclo[2.2.2]oct-2-ene (DBO) by nucleotides has been studied. The quenching mechanism was analyzed on the basis of deuterium isotope effects, tendencies for exciplex formation, and the quenching efficiency in the presence of a molecular container (cucurbit[7]uril). Exciplex-induced quenching appears to prevail for adenosine, cytidine, and uridine, while hydrogen abstraction becomes competitive for thymidine and guanosine. Compared to other fluorescent probes, DBO responds very selectively to the type of nucleotide.


De novo in vitro shoot morphogenesis from shoot tip-induced callus cultures of Gymnema sylvestre (Retz.) R.Br. ex Sm.
Gymnema sylvestre is a medicinal woody perennial vine known for its sweetening properties and anti-diabetic therapeutic uses in the modern and traditional medicines. Its over-exploitation for the therapeutic uses and to meet the demand of pharmaceutical industry in raw materials supply for the production of anti-diabetic drugs has led to considerable decline in its natural population. An efficient system of shoot bud sprouting from nodal segment explants and indirect plant regeneration from apical meristem-induced callus cultures of G. sylvestre have been developed on Murashige and Skoog (MS) medium amended with concentrations of cytokinins. Of the three growth regulators tested, N6-benzylaminopurine (BAP) was the most efficient and 2.0 mg L-1 gave the best shoot formation efficiency. This was followed by thidiazuron (TDZ) and kinetin (Kin) but, most of the TDZ-induced micro shoots showed stunted growth. Multiple shoot formation was observed on medium amended with BAP or TDZ at higher concentrations. The produced micro shoots were rooted on half strength MS medium amended with auxins and rooted plantlets acclimatized with 87% survival of the regenerates. The developed regeneration system can be exploited for genetic transformation studies, particularly when aimed at producing its high yielding cell lines for the anti-diabetic phytochemicals. It also offers opportunities for exploring the expression of totipotency in the anti-diabetic perennial vine.


Analysis of perfluoroalkyl substances in waters from Germany and Spain.
Water has been identified as one of the main routes of human exposure to perfluoroalkyl substances (PFASs). This work assessed the presence of 21 PFASs along the whole water cycle using a new fast and cost effective analytical method based on an online sample enrichment followed by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The method was validated for different types of matrices (ultrapure water, tap water and treated wastewater). The quality parameters for the 21 selected compounds presented good limits of detection (LOD) and quantification (LOQ) ranging, in general, from 0.83-10 ng/L to 2.8-50 ng/L, respectively. The method was applied to assess the occurrence of PFASs in 148 water samples of different steps along the whole water cycle, including: mineral bottled water, tap water, river water and treated effluent wastewater, from Germany to Spain. In addition, in order to prove the good performance of the online analytical method, the analysis of PFASs was carried out in parallel using a method based on offline anionic solid phase extraction (SPE) followed by LC-MS/MS. Consistent results were obtained using both approaches. The more frequently found compounds were perfluoroalkyl acids, such as the perfluorobutanoic acid which was in the 54% of the tap water samples investigated with concentrations in the range between 2.4 and 27 ng/L, the perfluoroheptanoic acid (0.23-53 ng/L) and perfluorooctanoic acid (0.16-35 ng/L), and the sulphonate perfluorooctanesulfonate (0.04-258 ng/L) which was the second more frequent compound and also the compound found in with the higher concentration. It should be remarked that the 88% of the samples analyzed presented at least one of the compounds at quantifiable concentrations. In addition, PFASs including short chain compounds were proved to be prevalent in drinking water, and the 50% of the drinking water samples showed quantifiable concentrations of PFASs. It should be said that the great majority of the samples may not pose an immediate health risk to consumers, and just 6 of the drinking water samples presented concentrations of PFOS exceeding the Provisional Health Advisory (PHA) level established by the Office of Water from the USEPA for PFOS, which was set in 200 ng/L.


Characterization of LipS1 and LipS2 from Thermococcus kodakarensis: Proteins Annotated as Biotin Synthases, which Together Catalyze Formation of the Lipoyl Cofactor.
Lipoic acid is an eight-carbon sulfur-containing biomolecule that functions primarily as a cofactor in several multienzyme complexes. It is biosynthesized as an attachment to a specific lysyl residue on one of the subunits of these multienzyme complexes. In Escherichia coli and many other organisms, this biosynthetic pathway involves two dedicated proteins: octanoyltransferase (LipB) and lipoyl synthase (LipA). LipB transfers an n-octanoyl chain from the octanoyl-acyl carrier protein to the target lysyl residue, and then, LipA attaches two sulfur atoms (one at C6 and one at C8) to give the final lipoyl cofactor. All classical lipoyl synthases (LSs) are radical S-adenosylmethionine (SAM) enzymes, which use an [Fe4S4] cluster to reductively cleave SAM to generate a 5'-deoxyadenosyl 5'-radical. Classical LSs also contain a second [Fe4S4] cluster that serves as the source of both appended sulfur atoms. Recently, a novel pathway for generating the lipoyl cofactor was reported. This pathway replaces the canonical LS with two proteins, LipS1 and LipS2, which act together to catalyze formation of the lipoyl cofactor. In this work, we further characterize LipS1 and LipS2 biochemically and spectroscopically. Although LipS1 and LipS2 were previously annotated as biotin synthases, we show that both proteins, unlike E. coli biotin synthase, contain two [Fe4S4] clusters. We identify the cluster ligands to both iron-sulfur clusters in both proteins and show that LipS2 acts only on an octanoyl-containing substrate, while LipS1 acts only on an 8-mercaptooctanoyl-containing substrate. Therefore, similarly to E. coli biotin synthase and in contrast to E. coli LipA, sulfur attachment takes place initially at the terminal carbon (C8) and then at the C6 methylene carbon.


[Epidemiological characteristics and analysis of related infection risk factors for influenza in Anhui Province from 2013 to 2021].
Objective: Analysis of the characteristics of influenza epidemic in Anhui Province and quantification of the impact of different factors on influenza occurrence, providing scientific basis for better influenza prevention and control. Methods: Descriptive analysis and factor analysis were conducted on influenza-like illness (ILI) cases and RT-PCR results in Anhui Province from 2013 to 2021 using data from China's Influenza Monitoring Information System. Results: The percentage of influenza-like illness (ILI%) of sentinel hospitals in Anhui Province from April 1, 2013 to March 31, 2021 was 3.80% (1 209 142/31 779 987), showing an overall increasing trend, with a relatively high proportion in 2017-2018 at 4.30% (191 148/4 448 211). The proportion of ILI cases in infants and young children aged 0-4 years was a relatively high at 54.14% (654 676/1 209 142), and the highest ILI% was observed in Fuyang City, Anhui Province (6.25%, 236 863/3 788 863). Laboratory monitoring results showed that the positive rate of ILI cases in sentinel hospitals in 8 influenza monitoring years was 16.38% (34 868/212 912), showing an increasing trend year by year, with a relatively proportion in 2017-2018 at 26.19% (6 936/26 488). The detection rate of school-age children aged 5-14 years was a relativelyhigh at 28.81% (13 869/48 144), and the positive rate was a relatively high in Wuhu City among the 16 cities, reaching 22.01% (2 693/122 237). Influenza activity showed a single peak in winter-spring and alternating double peaks in winter-spring and summer, with different subtypes alternating, and A (H3N2) was the dominant subtype in summer. The results of a multiple logistic regression model showed that the positive rate was higher in 2017-2018, among children aged 5-14 years, in winter, and in southern Anhui. Conclusions: Influenza epidemic in Anhui Province has a clear seasonal pattern, and the ILI% and detection rate have shown an upward trend from 2013 to 2021. Therefore, it is suggested to ensure vaccine supply before the winter-spring influenza season arrives, and to strengthen vaccine uptake and health education to avoid the risk of infection during the peak period of influenza.


[The incidence of Dicrocoelium dendriticum in Emmental].
A cross-sectional field study on the prevalence of Dicrocoelium dendriticum was performed in the Emmental. The study included 211 bovines, 170 equines, 20 ovines, 46 caprines and 23 rabbits (from 119 farms). In addition, laboratory routine diagnostic data obtained from 2.840 animals--all originating from the same area of investigation--were assessed in the same way. The infection extent concerning the different animal species were the following: bovines 46%, equines 12%, ovines 30%, caprines 48% and rabbits 9%. Univariate analyses of baseline epidemiological data identified no significant risk factors, with the exception of the type of stable used. Bovines kept in a modern free ranging stable had a significantly lower chance of infection with D. dendriticum than cattle in conventional tie stalls. The epidemiological data characterizing the area of investigation suggest the following procedure to reduce the problem of dicrocoeliosis: Pasturing animals of all ages should be regularly dewormed (e.g. every six week during pasture) using a compound effective against D. dendriticum. A treatment is especially indicated at the time after pasture in autumn or before housing the animals in winter. In spring, only animals having pastured the year before need to be treated prior to pasture in the new year. However, it is recommended to perform an economic analysis comparing costs of treatment versus putative costs of damage prior to the initiation of a strategic campaign: animal welfare aspects have to be considered. The laboratory routine diagnostic data showed infection extent similar to those of the cross-sectional study: bovines 60%, equines 24%, ovines 26%, caprines 31%, rabbits 32%. Atypical hosts such as dogs and cats exhibited low infection extent (3% and 1%, respectively), rather reflecting a gastro-intestinal passage of parasite eggs ingested by consumption of infected livers or by coprophagy of ruminant faeces.


Ohwia caudata aqueous extract attenuates doxorubicin-induced mitochondrial dysfunction in Wharton's jelly-derived mesenchymal stem cells.
Mitochondrial dysfunction has been linked to many diseases, including organ degeneration and cancer. Wharton's jelly-derived mesenchymal stem cells provide a valuable source for stem cell-based therapy and represent an emerging therapeutic approach for tissue regeneration. This study focused on screening the senomorphic properties of Ohwia caudata aqueous extract as an emerging strategy for preventing or treating mitochondrial dysfunction in stem cells. Wharton's jelly-derived mesenchymal stem cells were incubated with 0.1 μM doxorubicin, for 24 h to induce mitochondrial dysfunction. Next, the cells were treated with a series concentration of Ohwia caudata aqueous extract (25, 50, 100, and 200 μg/mL) for another 24 h. In addition, an untreated control group and a doxorubicin-induced mitochondrial dysfunction positive control group were maintained under the same conditions. Our data showed that Ohwia caudata aqueous extract markedly suppressed doxorubicin-induced mitochondrial dysfunction by increasing Tid1 and Tom20 expression, decreased reactive oxygen species production, and maintained mitochondrial membrane potential to promote mitochondrial stability. Ohwia caudata aqueous extract retained the stemness of Wharton's jelly-derived mesenchymal stem cells and reduced the apoptotic rate. These results indicate that Ohwia caudata aqueous extract protects Wharton's jelly-derived mesenchymal stem cells against doxorubicin-induced mitochondrial dysfunction and can potentially prevent mitochondrial dysfunction in other cells. This study provides new directions for the medical application of Ohwia caudata.


Biological activities of IC201 ((3S,8E)-1,3-dihydroxy-8-decen-5-one), a low molecular weight immunomodulator produced by Streptomyces.
IC201 was found in cultured broth of Streptomyces cirratus as an antitumor antibiotic which was effective in retarding growth of the established solid tumor of Ehrlich carcinoma by treatment starting 8 days after tumor inoculation. It retarded growth of the established solid tumor of IMC carcinoma but had no cytotoxicity at 100 micrograms/ml. IC201 treatment kept NK cell activity of tumor-bearing mice at normal level and stimulated cytostatic activity of peritoneal macrophages. It stimulated phagocytosis of yeast and phorbol myristate acetate-elicited superoxide production by peritoneal macrophages. The addition of IC201 to P388D1 cell cultures enhanced release of interleukin 1 (IL-1) into cultured supernatant but it affected lipopolysaccharide-induced IL-1 production. Although the addition to macrophage-depleted cultures did not show any stimulatory effect, mixed lymphocyte culture reaction was augmented in cultures using spleen cells as stimulator cells taken from mice given IC201. Results indicate that IC201 primarily activates macrophages and the activation may cause modulation of immune responses.


Experimental and clinical studies on plasma leptin in obese dogs.
Leptin is a protein synthesized and secreted primarily by adipocytes, and the circulating leptin concentration is elevated in obese humans and rodents. Recently, we have established a sandwich enzyme-linked immunosorbent assay for canine leptin. In the present study, plasma leptin concentrations were measured in experimentally developed obese beagles and in clinically obese dogs. When 5 male beagles were given a high-energy diet for 3 months, all of them became obese and the plasma leptin concentration significantly increased from 2.4+/-1.2 to 4.9+/-0.9 ng/ml, positively correlating with body fat content estimated by the deuterium oxide dilution method (r=0.87). The leptin concentrations of plasma samples collected from 59 dogs in veterinary practices were compared with their body condition scores (BCS). The plasma leptin concentrations of obese dogs were 9.7+/-0.7 and 12.3+/-1.5 ng/ml at BCS=4 and BCS=5, respectively, which were significantly higher than those of optimal (BCS=3) dogs (2.7+/-0.3 ng/ml). There was no significant effect of sex and breed. A weak positive correlation (r=0.37) was found between the plasma leptin concentration and age, probably due to the lesser content of visceral fat in puppies younger than 1 year old. These results indicate that plasma leptin is a good index of adiposity in dogs regardless of breed, age and sex, and may be useful for quantitative assessment of obesity in small animal practice.