Case Report: A Pseudoaneurysm Involving the Left Common Iliac Artery Secondary to Brucellosis: A Rare Case Report.
Pseudoaneurysms of the common iliac artery caused by Brucellosis are exceedingly uncommon. Infected common iliac artery pseudoaneurysms, particularly those caused by brucellosis, are more difficult to diagnose and cure than general pseudoaneurysms. The risk of mortality is significantly high in this condition. Nonsurgical treatment of a brucellosis-induced common iliac artery pseudoaneurysm is futile, and it should be operated on as soon as feasible. Long-term and multi-course antibacterial therapy with combination antibiotics is required. For the treatment of Brucella-infected pseudoaneurysms, endovascular surgery can be both effective and safe.


Intermittent positive airway pressure by nasal mask as a treatment for respiratory insufficiency in a patient with syringomyelia.
We report a case of a 73-year-old woman with chronic respiratory failure in association with syringohydromyelia and sleep apnea. She was initially intubated and mechanically ventilated. Weaning from the ventilator was only successful after nasal intermittent positive airway pressure therapy was started. A normalization of arterial blood gas exchange could be achieved.


In vitro antifungal activities of longan (Dimocarpus longan Lour.) seed extract.
Longan, Dimocarpus longan Lour., contains polyphenolic compounds which exhibit several pharmacological properties. This study aims to evaluate antifungal activities of longan fruit extract in comparison to its active compounds. The results showed that longan seed exhibited antifungal activity against the opportunistic yeasts (Candida species and Cryptococcus neoformans). In contrast, longan pulp and whole fruit did not demonstrate any inhibitory effects. Ellagic acid showed the most potent antifungal activity followed by corilagin and gallic acid, respectively. Ellagic acid inhibited Candida parapsilosis and C. neoformans more effectively than Candida krusei and also some Candida albicans clinical strains. Baidam cultivar possessed higher antifungal activity (MIC=500-4000 μg/ml) as it contained higher contents of ellagic acid and gallic acid than Edor (MIC=1000-8000 μg/ml). For antibacterial activity, only corilagin and gallic acid possessed weak to moderate inhibitory effects against Staphylococcus aureus and Streptococcus mutans, respectively. Longan seed was then applied in the oral care products. Longan effervescent granule (5% extract) significantly reduced adhesion of C. albicans to acrylic strips. Mouthwash containing 0.5% extract exhibited good antifungal activity compared to a commercial product. These findings indicated that longan seed extract and its polyphenolic compounds can be used as an antifungal agent in oral care products for the treatment of opportunistic yeast infection.


Assessment of Silicone Oil Emulsification: A Comparison of Currently Applied Methods.
To compare ultrasound biomicroscopy (UBM), Coulter counter, and B-scan ultrasonography in the evaluation of silicone oil (SO) emulsification. Patients who underwent primary pars plana vitrectomy with SO tamponade for rhegmatogenous retinal detachment and SO removal were included. UBM images were acquired before the SO removal, and B-scan images were taken after removal. The number of droplets in the first and last 2 mL of washout fluid was analyzed using a Coulter counter. The correlations between these measurements were analyzed. Thirty-four eyes received both UBM and Coulter counter analysis for the first 2 mL of washout fluid, and 34 underwent B-scan and Coulter counter analysis of the last 2 mL washout fluid. The mean UBM grading was 26.41 ± 9.71 (range: 1-36); the mean SO index obtained with B-scan was 5.25 ± 5.00% (range: 0.10-16.49%), and the mean number of SO droplets was 1.26 ± 2.45 × 107/mL and 3.34 ± 4.22 × 106/mL in the first and last 2 mL of washout fluid, respectively. There were significant correlations between UBM grading and SO droplets in the first 2 mL and between B-scan grading and SO droplets in the last 2 mL (all P < 0.05). UBM, Coulter counter, and B-scan ultrasonography could all be used in the evaluation of SO emulsification, and their findings were comparable.


Informal Caregiving Among American Indians and Alaska Natives in the Pacific Northwest.
Unpaid or informal caregivers are people who provide assistance without compensation to adults and children who require care beyond typical needs. Although often rewarding, informal caregiving can be associated with high rates of depression, stress, anxiety, sleep and endocrine system disruption, immunosuppression, and general morbidity and mortality. Few recent studies of informal caregivers have included data from American Indians and Alaska Natives (AI/AN). Given this noteworthy gap in the literature, we surveyed a total of 225 AI/ANs attending two cultural, community functions in the Pacific Northwest to gain a general understanding of frequency of caregiving, caregiver and recipient characteristics, caregiving duties, support needs, and financial, emotional, and physical strains as a consequence of caregiving. Of the 225 participants who completed the survey, 90 (40%) indicated that they had been an unpaid caregiver for a month or more and 28 (12%) were current unpaid caregivers. Consistent with prior research, most caregivers (84%) reported satisfaction from providing help, but 84% of caregivers reported experiencing "increased stress," 40% reported financial strain, and 34% reported decreased health "because of involvement with providing care." Our data also suggested a disproportionate impact on AI/AN women given higher rates of being a caregiver compared to other populations and less support from others in their communities. Our data suggest similarities for AI/AN caregivers with other groups of caregivers but also emphasize the importance of including AI/AN populations in future research in order to understand ways to best serve their unique needs.


Boom-Bust Turnovers of Megabase-Sized Centromeric DNA in Solanum Species: Rapid Evolution of DNA Sequences Associated with Centromeres.
Centromeres are composed of long arrays of satellite repeats in most multicellular eukaryotes investigated to date. The satellite repeat-based centromeres are believed to have evolved from "neocentromeres" that originally contained only single- or low-copy sequences. However, the emergence and evolution of the satellite repeats in centromeres has been elusive. Potato (Solanum tuberosum) provides a model system for studying centromere evolution because each of its 12 centromeres contains distinct DNA sequences, allowing comparative analysis of homoeologous centromeres from related species. We conducted genome-wide analysis of the centromeric sequences in Solanum verrucosum, a wild species closely related to potato. Unambiguous homoeologous centromeric sequences were detected in only a single centromere (Cen9) between the two species. Four centromeres (Cen2, Cen4, Cen7, and Cen10) in S. verrucosum contained distinct satellite repeats that were amplified from retrotransposon-related sequences. Strikingly, the same four centromeres in potato contain either different satellite repeats (Cen2 and Cen7) or exclusively single- and low-copy sequences (Cen4 and Cen10). Our sequence comparison of five homoeologous centromeres in two Solanum species reveals rapid divergence of centromeric sequences among closely related species. We propose that centromeric satellite repeats undergo boom-bust cycles before a favorable repeat is fixed in the population.


Several glycolipid fractions have been isolated from the bound lipids of N. brasiliensis. The main fraction was characterized as a monoester of nocardomycolic acids and D-arabinose. Fractionation of the free lipids yielded a glycolipid fraction consisting of nocardomycouc acids, arabinose, galactose and several amino acids. This last fraction appears to be similar to wax D of Mycobacteria; mild acid treatment of this fraction produced arabinose mononorcardomycolate.


Completion pericardiectomy for recurrent constrictive pericarditis: importance of timing of recurrence on late clinical outcome of operation.
Recurrent right-side heart failure after operation for constrictive pericarditis (CP) may be caused by incomplete pericardiectomy, recurrent constriction due to exuberant scar tissue, or diastolic dysfunction. Because the risks and benefits of reoperation are not well defined, we reviewed the outcome of completion pericardiectomy. From 1993 to December 2010, 41 patients underwent redo pericardiectomy. Thirty-eight patients had the initial operation elsewhere, and 3 had first pericardiectomy at our clinic. All patients had the diagnosis of CP at initial operation. We divided patients into two groups according to the interval between the first and second pericardiectomies: group A, 1 year or less, n = 20; group B, more than 1 year, n = 21. The mean age was 57.6 ± 12.7 years, and there were 34 males (83%). Twenty-six patients (63%) were in New York Heart Association class II, 10 (24%) were in class III, and 5 (12%) were in class IV. Etiologies were idiopathic in 20 (49%), prior cardiac surgery in 13 (32%), radiation in 6 (15%), and trauma in 2 (5%). There was no significant difference in patient characteristics between group A and group B. The 30-day and in-hospital mortalities were 7% (n = 3) and 12% (n = 5), respectively. Overall 5-year survival was 49%, and was significantly better in group A than group B (73% versus 29%, p = 0.032). In multivariate analysis, New York Heart Association class III or IV and the interval between operations longer than 1 year were significant risk factors for death (p = 0.010 and p = 0.027, respectively). The significant early mortality of repeat pericardiectomy emphasizes the importance of complete pericardial resection at first operation and accurate diagnosis of recurrent constriction. The poor clinical outcome of late (more than 1 year) reoperation suggests that many of these patients may have unrecognized diastolic dysfunction or recurrent mediastinal scarring as the cause of right-side heart failure rather than incomplete initial pericardiectomy.


SARS-CoV-2 infection in children evaluated in an ambulatory setting during Delta and Omicron time periods.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants and re-emergence of other respiratory viruses highlight the need to understand the presentation of and factors associated with SARS-CoV-2 in pediatric populations over time. The objective of this study was to evaluate the sociodemographic characteristics, symptoms, and epidemiological risk factors associated with ambulatory SARS-CoV-2 infection in children and determine if factors differ by variant type. We conducted a retrospective cohort study of outpatient children undergoing SARS-CoV-2 polymerase chain reaction testing between November 2020 and January 2022. Test-positive were compared with test-negative children to evaluate symptoms, exposure risk, demographics, and comparisons between Omicron, Delta, and pre-Delta time periods. Among 2264 encounters, 361 (15.9%) were positive for SARS-CoV-2. The cohort was predominantly Hispanic (51%), 5-11 years (44%), and 53% male; 5% had received two coronavirus disease 2019 (COVID-19) vaccine doses. Factors associated with a positive test include loss of taste/smell (adjusted odds ratio [aOR]: 6.71, [95% confidence interval, CI: 2.99-15.08]), new cough (aOR: 2.38, [95% CI: 1.69-3.36]), headache (aOR: 1.90, [95% CI: 1.28-2.81), fever (aOR: 1.83, [95% CI: 1.29-2.60]), contact with a positive case (aOR: 5.12, [95% CI: 3.75-6.97]), or household contact (aOR: 2.66, [95% CI: 1.96-3.62]). Among positive children, loss of taste/smell was more predominant during the Delta versus Omicron and pre-Delta periods (12% vs. 2% and 3%, respectively, p = 0.0017), cough predominated during Delta/Omicron periods more than the pre-Delta period (69% and 65% vs. 41%, p = 0.0002), and there were more asymptomatic children in the pre-Delta period (30% vs. 18% and 10%, p = 0.0023). These findings demonstrate that the presentation of COVID-19 in children and most susceptible age groups has changed over time.


Internal transcribed spacer polymorphism in Pseudo-nitzschia multistriata (Bacillariophyceae) in the Gulf of Naples: recent divergence or intraspecific hybridization?
The planktonic diatom Pseudo-nitzschia multistriata is a potentially toxic species recorded during late summer-autumn in the Gulf of Naples (Tyrrhenian Sea, Italy). We analysed the genetic structure by amplifying the internal transcribed spacer (ITS-1-5.8S-ITS-2) region of the ribosomal DNA of 44 strains isolated along 2 years. Polymorphism in the ITS region was detected by direct sequencing and the PCR-products from selected strains were thus cloned to assess intra-strain ITS variability. Strains clustered into three main types: type A and B - differing by 0.6% sequence divergence - and type A/B, showing both A and B variants within the same genome. The three types showed no differences in the large subunit sequences (LSU) of the rDNA, ultrastructure of the valve, secondary structure of ITS-1 and ITS-2, ploidy level and they were sexually compatible. Based on the results of these multiple approaches, we can state that the three ITS-types belong to the same reproductive unit (or "species" sensu Mayr [(1942). Systematics and the Origin of Species. Columbia University Press, New York]). We suggest that ITS polymorphism in P. multistriata may be related to the contemporary occurrence of different but still inbreeding populations which either diverged recently or originated in different geographic areas and became sympatric in the studied area.


Is it possible to stop follow-up of patients with primary T1G3 urothelial carcinoma of the bladder managed with intravesical bacille Calmette-Guérin immunotherapy?
Recurrence and progression of T1 grade 3 (T1G3) urothelial bladder carcinomas (UBCs) treated with bacille Calmette-Guérin (BCG) are common events, but the long-term follow-up of the disease remains controversial. To evaluate the long-term outcomes of BCG intravesical therapy in relation to disease recurrence and progression in primary T1G3 UBCs and upper tract disease. A single-institution, retrospective, population-based analysis of 316 patients with primary T1G3 UBC treated with transurethral resection (TUR) and BCG induction intravesical instillations was performed. Response was determined and monitored by routine periodic urine cytology, cystoscopy, and upper tract imaging. The median follow-up was 70 months (maximum 210 months). Among all of the tumours, 49.4 % did not relapse, 48.7 % recurred in the bladder during the first 5 years of surveillance, and only 6 patients (1.9 %) recurred after being free of disease during the first 5 years of follow-up. Nineteen percentage of the UBCs progressed to stage T2, and only 2 patients (1.2 %) progressed after the first 5 years of surveillance. An upper urinary tract recurrence was detected in 9.2 % of the patients; 65.5 % were diagnosed within the upper urinary tract during the first 5 years of follow-up. Following a 5-year tumour-free period, there is minimal risk of recurrence and progression in T1G3 UBCs treated with TUR and BCG induction intravesical instillations. This finding supports a less intensive and potentially less invasive surveillance scheme of bladder follow-up and upper urinary tract imaging in patients without any recurrence.


The antidysrhythmic effect of metoprolol in the epinephrine treated chick embryo.
It was confirmed through electrocardiography that within two hours after epinephrine treatment, four day chick embryos either maintained normal rhythm or developed a severe cardiac dysrhythmia (22/93, 24% dysrhythmic). The ECG dysrhythmia in epinephrine treated embryos were characterized by periods of bradycardia, asystole, and various supraventricular or ventricular dysrhythmias. Within four hours after treatment, dysrhythmic embryos either reestablished normal rhythmicity or died. Electrocardiographic data also demonstrated that metoprolol pretreatment will block epinephrine induced dysrhythmias (0/46, 0% dysrhythmic). We conclude that metoprolol possesses antidysrhythmic properties in the epinephrine treated chick embryo.


Molecular imaging and fusion targeted biopsy of the prostate.
This paper provides a review on molecular imaging with positron emission tomography (PET) and magnetic resonance imaging (MRI) for prostate cancer detection and its applications in fusion targeted biopsy of the prostate. Literature search was performed through the PubMed database using the keywords "prostate cancer", "MRI/ultrasound fusion", "molecular imaging", and "targeted biopsy". Estimates in autopsy studies indicate that 50% of men older than 50 years of age have prostate cancer. Systematic transrectal ultrasound (TRUS) guided prostate biopsy is considered the standard method for prostate cancer detection and has a significant sampling error and a low sensitivity. Molecular imaging technology and new biopsy approaches are emerging to improve the detection of prostate cancer. Molecular imaging with PET and MRI shows promising results in the early detection of prostate cancer. MRI/TRUS fusion targeted biopsy has become a new clinical standard for the diagnosis of prostate cancer. PET molecular image-directed, three-dimensional ultrasound-guided biopsy is a new technology that has great potential for improving prostate cancer detection rate and for distinguishing aggressive prostate cancer from indolent disease. Molecular imaging and fusion targeted biopsy are active research areas in prostate cancer research.


ADVANCE system testing: Benefit-risk analysis of a marketed vaccine using multi-criteria decision analysis and individual-level state transition modelling.
The Accelerated Development of VAccine beNefit-risk Collaboration in Europe (ADVANCE) is a public-private collaboration aiming to develop and test a system for rapid benefit-risk (B/R) monitoring of vaccines using electronic health record (eHR) databases in Europe. Proof-of-concept studies were designed to assess the proposed processes and system for generating the required evidence to perform B/R assessment and near-real time monitoring of vaccines. We aimed to test B/R methodologies for vaccines, using the comparison of the B/R profiles of whole-cell (wP) and acellular pertussis (aP) vaccine formulations in children as an example. We used multi-criteria decision analysis (MCDA) to structure the B/R assessment combined with individual-level state transition modelling to build the B/R effects table. In the state transition model, we simulated the number of events in two hypothetical cohorts of 1 million children followed from first pertussis dose till pre-school-entry booster (or six years of age, whichever occurred first), with one cohort receiving wP, and the other aP. The benefits were reductions in pertussis incidence and complications. The risks were increased incidences of febrile convulsions, fever, hypotonic-hyporesponsive episodes, injection-site reactions and persistent crying. Most model parameters were informed by estimates (coverage, background incidences, relative risks) from eHR databases from Denmark (SSI), Spain (BIFAP and SIDIAP), Italy (Pedianet) and the UK (RCGP-RSC and THIN). Preferences were elicited from clinical and epidemiological experts. Using state transition modelling to build the B/R effects table facilitated the comparison of different vaccine effects (e.g. immediate vaccine risks vs long-term vaccine benefits). Estimates from eHR databases could be used to inform the simulation model. The model results could be easily combined with preference weights to obtain B/R scores. Existing B/R methodology, modelling and estimates from eHR databases can be successfully used for B/R assessment of vaccines.


Pure ground glass opacity (GGO) on chest CT: a rare presentation of lung metastasis of Malignant Phyllodes Tumor.
Lung is the most common target organ for distant metastasis of phyllodes tumor (PT), where the metastatic tumors are mostly seen as nodules or masses. We report here a rare case in which pure ground-glass opacity (GGO) was observed on chest CT about 3 years after the initial treatment of breast PT. After the lung lobectomy, we histologically confirmed that it should be diagnosed as a metastasis of PT. GGO rarely shows on metastatic tumors, and no reports have been made on GGO thus far in connection with metastasis of PT. Thus, in case GGO was found on the CT of patients having a history of PT, we suggest to take possible distant-metastasis into consideration.


Strongyloides ransomi: proteolytic enzymes from larvae.
The filariform larvae of Strongyloides ransomi can infect their hosts by penetration through skin. In this report, homogenates of these organisms were prepared and their proteolytic enzymes examined. Homogenates prepared in 0.2 M citrate, pH 4.0, contain two thiol-dependent proteinases with molecular weights of approximately 32,000 and 28,000. These proteinases have an acidic pH optimum and show substrate preferences and inhibitor susceptibilities similar to the vertebrate acidic cysteinyl proteinases. Homogenates prepared in 0.1 M Tris, pH 7.5, contain multiple proteolytic enzymes, active against both Azocoll and synthetic substrates. These enzymes do not require thiols for activity and they have an alkaline pH optimum. The enzymes are inhibited by both chelating agents and heavy metals, but not by serine-proteinase inhibitors. Extracts prepared in 0.1 M Tris-HCl, pH 7.5, contain endogenous proteinase inhibitors.


When Jump Height is not a Good Indicator of Lower Limb Maximal Power Output: Theoretical Demonstration, Experimental Evidence and Practical Solutions.
Lower limb external maximal power output capacity is a key physical component of performance in many sports. During squat jump and countermovement jump tests, athletes produce high amounts of mechanical work over a short duration to displace their body mass (i.e. the dimension of mechanical power). Thus, jump height has been frequently used by the sports science and medicine communities as an indicator of the power output produced during the jump and by extension, of maximal power output capacity. However, in this article, we contend that squat jump and countermovement jump height are not systematically good indicators of power output produced during the jump and maximal power output capacity. To support our opinion, we first detail why, theoretically, jump height and maximal power output capacity are not fully related. Specifically, we demonstrate that individual body mass, push-off distance, optimal loading and the force-velocity profile confound the jump height-power relationship. We also discuss the relationship between squat jump or countermovement jump height and maximal power output capacity measured with a force plate based on data reported in the literature, which added to our own experimental evidence. Finally, we discuss the limitations of existing practical solutions (regression-based estimation equations and allometric scaling), and advocate using a valid, reliable and simple field-based procedure to compute individual power output produced during the jump and maximal power output capacity directly from jump height, body mass and push-off distance. The latter may allow researchers and practitioners to reduce bias in their assessment of lower limb mechanical power output by using jump height as an input with a simple yet accurate computation method, and not as the first/only variable of interest.


Assessment of human resources for health programme implementation in 15 Latin American and Caribbean countries.
The health systems in the Americas region are characterized by fragmentation and segmentation, which constitute an important barrier to expanding coverage, achieving integrated primary health care, and reducing inefficiency and discontinuity of care. An assessment of the human resources for health (HRH) programmes that have been implemented at the country level was developed as part of the measurement of the 20 HRH regional goals for 2007-2015, adopted in 2007 by the Pan American Sanitary Conference (CSPA). The exercise was a combination of academic research and the development/application of an advocacy tool involving policy makers and stakeholders to influence the decision-making in the development, implementation, or change of HRH programmes while building evidence through a structured approach based on qualitative and quantitative information and the exchange and dissemination of best practices. This paper covers the methodological challenges, as well as a summary of the main findings of the study, which included 15 countries: Belize, Costa Rica, El Salvador, Guatemala, Honduras, Nicaragua and Panama in the Central America, Dominican Republic in the Caribbean, Chile, Colombia, Ecuador and Peru in the Andean sub region, and Argentina, Paraguay, and Uruguay in the South Cone. Despite the different contexts, the results showed that the programmes evaluated faced common challenges, such as lack of political support and financial unsustainability. The evaluation process allowed the exchange and dissemination of practices, interventions, and programmes currently running in the region. A shared lesson was the importance of careful planning of the implementation of programmes and interventions. The similarities in the problems and challenges of HRH among the participating countries highlighted the need for a cooperation programme on the evaluation and assessment of implementation strategies in the Americas region.


Determination of ochratoxin A at part-per-trillion level in Italian salami by immunoaffinity clean-up and high-performance liquid chromatography with fluorescence detection.
A fast high-performance liquid chromatography method has been devised for the determination of ochratoxin A (OTA) in Italian salami in the low part-per-trillion (pg/g) level. The samples were extracted with ethyl acetate and purified by immunoaffinity column (IAC). The IAC eluate could be directly injected or previously concentrated 10-fold. Recovery at 0.5 and 1 ng/g was 77 +/- 4%. The between-day coefficient of variation measured over 5 days on samples spiked at 1 ng/g was 8%. The developed method required a relatively small volume of non-halogenated organic solvent and the whole procedure was simpler and faster compared to other existing procedures. The limit of detection was 0.06 ng/g that could be even lowered using a preconcentration step. A total of 30 salami samples were analysed using this procedure; the most contaminated sample was found to have OTA concentration at 0.4 ng/g level.


[Retinal Detachment Part 2 - Treatment Strategies].
This review article summarizes the relevant surgical strategies for retinal detachment repair and discusses common postoperative complications, as well as factors influencing the functional and anatomical results. Treatment of retinal detachment requires surgery. In most cases, pars plana vitrectomy is performed. In phakic patients with a clear lens scleral buckling can be considered as an alternative. Laser and cryotherapy are still the standard for treatment in symptomatic retinal tears or degenerations without retinal detachment. Postoperatively, the occurrence of a re-detachment as well as the development of proliferative vitreoretinopathy can have a negative influence on postoperative results. Furthermore, typical complications include the occurrence of a cystoid macular edema, epiretinal membrane or even persistent neurosensory detachment. Functional results are significantly influenced by macular involvement and the primary anatomical success rate.


Lowe syndrome identified in the offspring of an oocyte donor who was an unknown carrier of a de novo mutation: a case report and review of the literature.
Oculocerebrorenal syndrome of Lowe is an X-linked disorder with very low prevalence in the general population. The OCRL gene encodes the protein phosphatidylinositol 4,5-bisphosphate-5-phosphatase, a lipid phosphatase, located in the trans-Golgi network. Point mutations in the OCRL gene cause Lowe syndrome and Dent disease, which are characterized as a multisystemic disorder. The symptoms of Lowe syndrome are expressed primarily as dysfunction of the eyes, kidneys, and the central nervous system. This report describes a case of a 31-year-old Georgian woman with a de novo pathogenic mutation causing oculocerebrorenal syndrome of Lowe, who was a volunteer in an oocyte donation program for in vitro fertilization purposes, and the outcome of the treatments of this particular donor's oocyte receivers, describing the implications of the mutation for the children born as a result of the treatments. It raises important medical and ethical issues about the necessity of genetic testing of oocyte donors and the possibility of rare genetic disorders being inherited by the offspring of donors. This particular case indicates the legal, medical, and emotional risks of utilizing donor oocytes from phenotypically healthy women, whose genetic constitution is unknown in terms of being silent carriers of rare diseases. In addition, all the necessary actions were followed; the further examinations that are required are mentioned. The donor and the offspring should be further tested. The remaining cryopreserved embryos should be destroyed or preimplantation genetic testing should be performed before they are utilized. Finally, all the people involved, the treated couples and the donor, alongside her family, should follow genetic and psychological counselling.


On the origin, evolution, and nature of programmed cell death: a timeline of four billion years.
Programmed cell death is a genetically regulated process of cell suicide that is central to the development, homeostasis and integrity of multicellular organisms. Conversely, the dysregulation of mechanisms controlling cell suicide plays a role in the pathogenesis of a wide range of diseases. While great progress has been achieved in the unveiling of the molecular mechanisms of programmed cell death, a new level of complexity, with important therapeutic implications, has begun to emerge, suggesting (i) that several different self-destruction pathways may exist and operate in parallel in our cells, and (ii) that molecular effectors of cell suicide may also perform other functions unrelated to cell death induction and crucial to cell survival. In this review, I will argue that this new level of complexity, implying that there may be no such thing as a 'bona fide' genetic death program in our cells, might be better understood when considered in an evolutionary context. And a new view of the regulated cell suicide pathways emerges when one attempts to ask the question of when and how they may have become selected during evolution, at the level of ancestral single-celled organisms.


Structural and spectroscopic diagnosis of Eu:ZnO and Eu:Yb:ZnO glass and ceramics.
Eu and Eu:Yb codoped ZnO nanocrystals embedded in B(2)O(3) glass matrix were synthesized and their structural, optical properties were discussed. On excitation with 532 nm laser radiation intense orange/red emissions from Eu(3+) ions were observed. The fluorescence intensity was found to enhance on annealing. Frequency upconversion emissions from Eu(3+) ions were observed in presence of Yb(3+) ions on excitation with NIR (976 nm) laser. The enhancement in emission intensity was explained and the photo-physics involved is linked with the unique structural properties of the crystallites formed. Ion interactions and the different energy transfer parameters were also calculated.


The mycobacteriology of pulmonary tuberculosis in South African gold miners.
Two bacteriological surveys of gold miners with pulmonary tuberculosis diagnosed for the first time, have shown a stable level of primary drug resistance which is substantially lower than that reported for the home areas of these men. Initial drug resistance was detected in 12.7% of the 205 cultures of Mycobacterium tuberculosis in 1983-1984 and in 10.7% of 253 cultures in 1988-1989. Resistance to isoniazid was detected in 5.4% and 5.5% of the strains tested, to streptomycin in 6.8% and 5.1% and to rifampicin in 0% and 0.8% respectively. In a separate study, the sputum bacteriology of 691 miners with relapsed pulmonary tuberculosis was examined. M. tuberculosis was cultured from the sputum of 547 of the men and environmental mycobacteria (EM) in the remainder. Drug resistant tuberculosis was found in 20.5% of the previously treated men: isoniazid resistance in 14.3%, streptomycin resistance in 11.2% and rifampicin resistance in 3.7%. All of the rifampicin resistant strains were also resistant to isoniazid and 85% were resistant to streptomycin as well. Only EM were cultured from 29 of the 234, 1983-1984 sputum cultures, from 23 of the 276, 1988-1989 cultures and from 144 of the 691 relapse cultures. Mycobacterium kansasii (67%) was the commonest EM cultured.


Nature of toxicity for chick embryo fibroblast cells of coumermycin A1 and its physico-chemical interactions with protein and nucleic acid.
The results reported in this paper describe the effects produced by the antibiotic Coumermycin A1 (CA1) on survival and metabolism of chick embryo fibroblast cells (CEF), and give a clue to the understanding of its toxicity. The drug acts primarily at the level of DNA and RNA synthetic enzymes; no effect on DNA superstructure is detectable at doses at which cytotoxicity is pronounced. A spectroscopic approach produced evidence that CA1 binds to DNA, RNA, chromatin components such as histones and to a structurally unrelated protein such as bovine serum albumin. Furthermore, CA1 behaves like a pure non-competitive inhibitor of lactic dehydrogenase, a ubiquitous enzyme not involved in nucleic acid metabolism. The interaction of CA1 with a wide range of macromolecules playing different biological roles is certainly relevant to its activity and adds a new insight into the mechanism of action of this antibiotic. These observations are also discussed in the light of the alleged role of CA1 as a specific inhibitor of DNA topoisomerase in eukaryotic cells.


Intravenously administered APAC, a dual AntiPlatelet AntiCoagulant, targets arterial injury site to inhibit platelet thrombus formation and tissue factor activity in mice.
Arterial thrombosis is the main underlying mechanism of acute atherothrombosis. Combined antiplatelet and anticoagulant regimens prevent thrombosis but increase bleeding rates. Mast cell-derived heparin proteoglycans have local antithrombotic properties, and their semisynthetic dual AntiPlatelet and AntiCoagulant (APAC) mimetic may provide a new efficacious and safe tool for arterial thrombosis. We investigated the in vivo impact of intravenous APAC (0.3-0.5 mg/kg; doses chosen according to pharmacokinetic studies) in two mouse models of arterial thrombosis and the in vitro actions in mouse platelets and plasma. Platelet function and coagulation were studied with light transmission aggregometry and clotting times. Carotid arterial thrombosis was induced either by photochemical injury or surgically exposing vascular collagen after infusion of APAC, UFH or vehicle. Time to occlusion, targeting of APAC to the vascular injury site and platelet deposition on these sites were assessed by intra-vital imaging. Tissue factor activity (TF) of the carotid artery and in plasma was captured. APAC inhibited platelet responsiveness to agonist stimulation (collagen and ADP) and prolonged APTT and thrombin time. After photochemical carotid injury, APAC-treatment prolonged times to occlusion in comparison with UFH or vehicle, and decreased TF both in carotid lysates and plasma. Upon binding from circulation to vascular collagen-exposing injury sites, APAC reduced the in situ platelet deposition. Intravenous APAC targets arterial injury sites to exert local dual antiplatelet and anticoagulant actions and attenuates thrombosis upon carotid injuries in mice. Systemic APAC provides local efficacy, highlighting APAC as a novel antithrombotic to reduce cardiovascular complications.


Epileptic encephalopathy with suppression-bursts and nonketotic hyperglycinemia.
Bursts of paroxysmal activity alternating with lack of activity define the suppression-burst (SB) pattern that may be acute, in hypoxic-ischemic encephalopathy and barbiturate intoxication, or chronic in the course of early epileptic and neonatal myoclonic (NME) encephalopathies. Malformations, namely Aicardi syndrome and hemimegalencephaly, gene mutations - of ARX and MUNC18 -, and inborn errors of metabolism, namely glycine encephalopathy, are the main causes, with spasms indicating more likely a malformation whereas myoclonus indicates metabolic disorders. Although glycine encephalopathy has a very severe outcome in its classical expression, it may be transient in the neonatal period, for reasons yet not identified. Although glycine encephalopathy is the main identified cause of NME, the disorder may not cause SB, especially in cases with later onset. The biochemical bases, due to changes in one of the four proteins that compose the enzyme, are well understood, but there is no phenotype-genotype correlation. Prenatal diagnosis is based on villous biopsy. The mechanism of SB partly depends on glutamate - or glycine, the co-neurotransmitter for NMDA transmission - overflow, mainly in the immature brain but also in cases due to barbiturate intoxication. Energy supply defect may also be involved in some inborn errors of metabolism.


Laparoscopic versus open reduction of intussusception in children: experience over a decade.
Intussusception is a common cause of bowel obstruction in children, which sometimes necessitates operative reduction and or resection. We report our series of patients with intussusception who were treated laparoscopically (LAP group) compared with exploratory laparotomy (OPEN group). After institutional review board approval, a retrospective review was performed evaluating outcomes for patients requiring surgical reduction of intussusception over a 10-year period. Analysis was based on intent to treat, and technique of exploration was surgeon's choice. Data were analyzed with the Wilcoxon rank sum test and chi-squared test where appropriate. P≤.05 was considered significant. During the time period studied, there were 92 patients treated surgically for intussusception: 65 LAP and 27 OPEN. Conversion to the open procedure was required for 21 patients in the LAP group, and of those, 6 required bowel resection. Seven of the patients who were started in the OPEN group ultimately required bowel resection. Operative time, length of hospital stay, time to full feeds, and total days of narcotics were all significantly shorter for the LAP group compared with the OPEN group (P=.003, P=.001, P=.001, and P=.004, respectively). A pathologic lead point was found in 14% of LAP and 15% of OPEN cases. In a subset analysis, 33% of patients who were converted from the LAP group to the open procedure had a pathologic lead point. Complication rates between the LAP and OPEN groups were comparable. Laparoscopy appears to be a safe and effective technique for reducing intussusception in children. The laparoscopic cases had shorter operative time, shorter time to full feeds, lower requirement for intravenous narcotics, and earlier discharges.


Synthesis and characterisation of four bimetallic gold-gallium clusters with Au-Ga rings as a new structural motif in gold cluster chemistry.
Using the newly introduced reducing agent in gold cluster chemistry GaCp, four new bimetallic gold-gallium cluster compounds were synthesised, stabilised by alkylphosphine ligands. The gold core of Au6(GaCl2)4(PEt3)6, Au7(GaCl2)3(P n Pr3)6 and Au13(GaCl2)5(GaCl)(P n Pr2 n Bu)9 is composed of two, three and four Au4 tetrahedra, linked in different ways. This arrangement gives the last-mentioned cluster the shape of a five-pointed star. A fourth synthesised metalloid gold cluster Au13(GaCl2)5(P n Pr3)9 consists of a voluminous gold core encircled by a gold-gallium chain. These gold-gallium chains are present in all four cluster compounds and are similar to the well-known gold-sulphur staple motifs in thiol-stabilised gold cluster compounds.


Influence of pyramidotomy on limb flexion movements induced by cortical stimulation and on associated postural adjustment in the cat.
Flexion movements induced by cortical stimulation and the associated postural adjustments in bilaterally pyramidotomised cats have been studied by means of an apparatus which measures separately the changes of force under each limb in the upright position of the animal. The results show: (1) The general motor behaviour of the animal is not affected by the lesion. The principal deficit is loss of tactile placing reaction in the forelimbs; there also appears a state akin to a vestibular syndrome when a light restraining harness is placed around the back and chest. (2) Stimulation of the motor cortex continues to evoke flexion movements of the contralateral limbs together with associated postural adjustment. Coordination between movement and postural adjustment is generally similar to that observed before pyramidal section, and response thresholds are unchanged. (3) Measurements reveal great quantitative modification both of the movement and the postural adjustment after the lesion. Weight shift latencies are increased and more dispersed in time, while the weight shifts themselves are reduced in amplitude and speed. (4) All these changes are more marked in the case of forelimb, rather than hindlimb, flexion, emphasising the greater degree of pyramidal influence on forelimb activity in the normal animal. (5) The results as a whole underline the ability of non-pyramidal systems, under the control of the motor cortex, to bring about both limb flexion and the associated postural adjustments in the absence of the corticospinal pathway.


Enantiomeric separation of citalopram and its metabolites by capillary electrophoresis.
A simple and fast capillary electrophoretic method has been developed for the enantioselective separation of citalopram and its main metabolites, namely N-desmethylcitalopram and N,N-didesmethylcitalopram, using beta-cyclodextrin (beta-CD) sulfate as the chiral selector. For method optimisation several parameters were investigated, such as CD and buffer concentration, buffer pH, and capillary temperature. Baseline enantioseparation of the racemic compounds was achieved in less than 6 min using a fused-silica capillary, filled with a background electrolyte consisting of a 35 mM phosphate buffer at pH 2.5 supplemented with 1% w/v beta-CD sulfate and 0.05% w/v beta-CD at 25 degrees C and applying a voltage of -20 kV. A fast separation method for citalopram was also optimized and applied to the analysis of pharmaceutical formulations. Racemic citalopram was resolved in its enantiomers in less than 1.5 min using short-end injection (8.5 cm, effective length) running the experiments in a background electrolyte composed of a 25 mM citrate buffer at pH 5.5 and 0.04% w/v beta-CD sulfate at a temperature of 10 degrees C.


EBP50 inhibits the anti-mitogenic action of the parathyroid hormone type 1 receptor in vascular smooth muscle cells.
Parathyroid hormone-related protein (PTHrP) and the parathyroid hormone type 1 receptor (PTH1R) are important regulators of vascular remodeling. PTHrP expression is associated to increased proliferation of vascular smooth muscle cells (VSMC). In contrast, signaling via the PTH1R inhibits cell growth. The mechanisms regulating the dual effect of PTHrP and PTH1R on VSMC proliferation are only partially understood. In this study we examined the role of the adaptor protein ezrin-radixin-moesin-binding phosphoprotein (EBP50) on PTH1R expression, trafficking, signaling and control of A10 cell proliferation. In normal rat vascular tissues, EBP50 was restricted to the endothelium with little expression in VSMC. EBP50 expression significantly increased in VSMC following angioplasty in parallel with PTHrP. Interestingly, PTHrP was able to induce EBP50 expression. In the clonal rat aortic smooth muscle cell line A10, EBP50 increased the recruitment of PTH1R to the cell membrane and delayed its internalization in response to PTHrP(1-36). This effect required an intact C-terminal motif in the PTH1R. In naïve A10 cells, PTHrP(1-36) stimulated cAMP production but not intracellular calcium release. In contrast, PTHrP(1-36) induced both cAMP and calcium signaling in A10 cells over-expressing EBP50. Finally, EBP50 attenuated the induction of p27(kip1) and the anti-proliferative effect of PTHrP(1-36). In summary, this study demonstrates the dynamic expression of EBP50 in vessels following injury and the effects of EBP50 on PTH1R function in VSMC. These findings highlight one of the mechanisms leading to increased VSMC proliferation and have important implication in the understanding of the molecular events leading to restenosis.


Effect of reperfusion temperature and pressure on the functional and metabolic recovery of preserved hearts.
Reperfusion conditions significantly affect recovery following global myocardial ischemia. Using an isolated dog heart model, we investigated the effect of initial (first 10 minutes) reperfusion temperature and pressure on the metabolic and functional recovery of the preserved heart. Four groups of five or six dogs each underwent 2 hours of ischemic cardioplegic arrest at 15 degrees C following single-dose crystalloid cardioplegia. Hearts were initially reperfused at 37 degrees C (high temperature) or 28 degrees C (low temperature) and at 50 mm Hg (low pressure) or 80 mm Hg (high pressure), giving four groups: (1) high-temperature, high-pressure; (2) high-temperature, low-pressure; (3) low-temperature, high-pressure; and (4) low-temperature, low-pressure. Septal temperatures were continuously recorded. Ventricular function curves 1 and 2 hours following reperfusion were significantly depressed in the high-temperature, high-pressure group (70%, p less than 0.01, and 83%, p less than 0.02) and the low-temperature, high-pressure group (78%, p less than 0.03, and 85%, p less than 0.03) but were normal in the low-temperature, low-pressure and the high-temperature, low-pressure groups. All groups showed edema 1/2 hour after reperfusion as measured by water and sodium content in myocardial biopsy specimens but only the high-temperature, high-pressure and the low-temperature, high-pressure groups showed persistent edema at 3 hours (3.95 +/- 0.2 ml H2O/gm dry weight, p less than 0.03 and 3.99 +/- 0.16 ml/gm, p less than 0.02, respectively). Only low-temperature, high-pressure reperfusion resulted in statistically significant reductions in adenosine triphosphate (ATP) 1/2 hour and 2 hours following reperfusion (a 15% reduction from baseline). Maximum rewarming rates were measured for each group. High-temperature, high-pressure = 2 degrees C per second; low-temperature, high-pressure = 1 degree C per second; high-temperature, low-pressure = 0.75 degrees C per second; and low-temperature, low-pressure = 0.4 degrees C per second. High-pressure reperfusion following global myocardial ischemia results in rapid rewarming and is associated with prolonged myocardial edema, depressed ATP levels, and delayed functional recovery. Therefore, we employ 10 minutes of low-pressure reperfusion in our patients undergoing potassium cardioplegic arrest.


Salvage Radical Prostatectomy after Primary Focal Ablative Therapy: A Systematic Review and Meta-Analysis.
Focal therapy (FT) has been gaining popularity as a treatment option for localized intermediate-risk prostate cancer (PCa) due to the associated lower morbidity compared to whole-gland treatment. However, there is an increased risk of local cancer recurrence requiring subsequent treatment in a small proportion of patients. To conduct a systematic review and meta-analysis to better describe and analyze patient postoperative, oncologic, and functional outcomes for those who underwent salvage radical prostatectomy (sRP) to manage their primary FT failure. A systematic review was completed using three databases (PubMed, Embase, and CINAHL) from October to December 2021 to identify data on outcomes in patients who received sRP for cancer recurrence after prior focal treatment. 12 articles (482 patients) were included. Median time to sRP was 24 months. Median follow-up time was 27 months. A meta-analysis revealed a postoperative complication rate of 15% (95% CI: 0.09, 0.24), with 4.6% meeting criteria for a major complication Clavien (CG) grade ≥3. Severe GU toxicity was seen in 3.6% of the patients, and no patients had severe GI toxicity. Positive surgical margins (PSM) were found in 27% (95% CI: 0.19, 0.37). Biochemical recurrence (BCR) after sRP occurred in 23% (95% CI: 0.17, 0.30), indicating a BCR-free probability of 77% at 2 years. Continence (pad-free) and potency (ability to have penetrative sex) were maintained in 67% (95% CI: 0.53, 0.78) and 37% (95% CI: 0.18, 0.62) at 12 months, respectively. Our evidence shows acceptable complication rates and oncologic outcomes; however, with suboptimal functional outcomes for patients undergoing sRP for recurrent PCa after prior FT. Inferior outcomes were observed for salvage treatment compared to primary radical prostatectomy (pRP). More high-quality studies are needed to better characterize outcomes after this sequence of PCa treatments. We looked at treatment outcomes and toxicity for men treated with sRP for prior FT failure. We conclude that these patients will have significant detriment to genitourinary function, with outcomes being worse than those for pRP patients.


Effectiveness of acupuncture as auxiliary combined with Western medicine for epilepsy: a systematic review and meta-analysis.
Although more and more clinical studies have shown that acupuncture as an auxiliary combined with Western medicine is effective in the treatment of patients with epilepsy, no systematic reviews of acupuncture as a treatment for epilepsy have been published. Hence, we conducted this meta-analysis to evaluate the effect of acupuncture treatment on patients with epilepsy. This study retrieved randomized controlled trials (RCTs) of acupuncture treatment for epilepsy from various electronic databases including PubMed, Embase, Cochrane Library, China National Knowledge Infrastructure, Chinese BioMedical Literature Database, and Wangfang database. These studies evaluated the effectiveness of acupuncture as an auxiliary treatment combined with Western medicine for patients with epilepsy. The methodological quality of the studies was assessed using the Cochrane Handbook for Systematic Reviews of Interventions. A total of 17 RCTs involving a total of 1,389 participants were included. The results showed that acupuncture combined with Western medicine improved the effective rates of treatment (OR: 4.28; 95% CI: 3.04-6.02; p < 0.001), and reduced the seizure frequency of patients (SMD: -3.29; 95% CI: -3.51 to -3.07; p < 0.001) and the EEG discharge frequency (SMD: -5.58; 95% CI: -7.02 to -4.14; p < 0.001). Regarding the quality of life and adverse events, the acupuncture group was superior to the control group in improving the overall quality of life of patients with epilepsy (SMD: 14.41; 95% CI: 12.51-16.32; p < 0.001) and decreased adverse events (OR: 0.38; 95% CI: 0.23-0.63, p < 0.001). The results of the analysis suggested that acupuncture combined with Western medicine is probably helpful in patients with epilepsy, but strong supportive data are not yet available. Given that this study is based on a low to moderate evidence-based analysis, the conclusions should be viewed with caution. PROSPERO, identifier no. CRD42023409923.


Neuropsychological Symptoms and Intrusive Thoughts Are Associated With Worse Trajectories of Chemotherapy-Induced Nausea.
Although chemotherapy-induced vomiting is well controlled with evidence-based antiemetic regimens, chemotherapy-induced nausea (CIN) remains a significant clinical problem. Study purposes, in a sample of outpatients with breast, gastrointestinal, gynecological, or lung cancer who received two cycles of chemotherapy (CTX, n = 1251), were to evaluate for interindividual differences in the severity of CIN and to determine which demographic, clinical, symptom, and stress characteristics are associated with higher initial levels as well as with the trajectories of CIN. Patients were recruited during their first or second cycle of CTX. Patients completed self-report questionnaires a total of six times over two cycles of CTX. Hierarchical linear modeling was used to evaluate for interindividual differences in and characteristics associated with the severity of CIN. Across the two cycles of CTX, higher levels of sleep disturbance, depression, and morning fatigue, as well as higher levels of intrusive thoughts, were associated with higher initial levels of CIN. In addition, lower functional status scores and shorter cycle lengths were associated with higher initial levels of CIN, and younger age and higher emetogenicity of the CTX regimen were associated with both higher initial levels as well as worse trajectories of CIN severity. These findings suggest that common symptoms associated with cancer and its treatment are associated with increased severity of CIN. Targeted interventions for these symptoms may reduce the burden of unrelieved CIN.


Age-related decrease in brain synaptic membrane Ca2+-ATPase in F344/BNF1 rats.
We used Fisher 344/Brown Norway hybrid rats (F344/BNF1) to determine whether previously reported decreases in brain synaptic plasma membrane (SPM) Ca2+-ATPase activity in inbred F344 rats also occurred in the hybrids. Plasma membrane Ca2+-ATPase (PMCA) activity in SPMs from F344/BNF1 rat brains showed a progressive age-dependent decrease in Vmax from 60.9 +/- 3.7 nmol Pi/mg/min (n = 6) in 5-month rats to 32.4 +/- 3.6 nmol Pi/mg/min (n = 6) in 34-month animals, with no change in K (act) for Ca2+. Immunoreactive PMCA in SPMs also decreased by approximately 20% at 34 months, and the calmodulin (CaM) bound to membranes following extraction with EDTA also declined progressively with age. The effectiveness of CaM in stimulating PMCA activity was significantly lower when CaM was purified from the brains of old vs. young F344 rats and when CaM from 5-month rats was oxidized in vitro. These results indicate: 1) that PMCA activity in SPMs from longer lived F344/BNF1 hybrids also decreases with age; 2) that part of the reduction in PMCA activity is due to loss of PMCA from the membranes; and 3) that age-related structural changes in CaM may decrease its interaction with proteins in SPMs.


Self-powered supercapacitive microbial fuel cell: The ultimate way of boosting and harvesting power.
In this work, for the first time, we demonstrate a supercapacitive microbial fuel cell which integrates the energy harvesting function of a microbial fuel cell (MFC) with the high-power operation of an internal supercapacitor. The pursued strategies are: (i) the increase of the cell voltage by the use of high potential cathodes like bilirubin oxidase (BOx) or iron-aminoantipyrine (Fe-AAPyr); (ii) the use of an additional capacitive electrode (additional electrode, AdE) which is short-circuited with the MFC cathode and coupled with the MFC anode (MFC-AdE). The high working potential of BOx cathode and the low impedances of the additional capacitive electrode and the MFC anode permitted to achieve up to 19 mW (84.4 Wm(-2), 152 Wm(-3)), the highest power value ever reported for MFCs. Exploiting the supercapacitive properties of the MFC electrodes allows the system to be simpler, cheaper and more efficient without additional electronics management added with respect to an MFC/external supercapacitor coupling. The use of the AdE makes it possible to decouple energy and power and to achieve recharge times in the order of few seconds making the system appealing for practical applications.


Frozen for combat: Quality of deep-frozen thrombocytes, produced and used by The Netherlands Armed Forces 2001-2021.
The Netherlands Armed Forces (NLAF) are using -80°C deep-frozen thrombocyte concentrate (DTC) since 2001. The aim of this study is to investigate the effect of storage duration and alterations in production/measurement techniques on DTC quality. It is expected that DTC quality is unaffected by storage duration and in compliance with the European guidelines for fresh and cryopreserved platelets. Pre-freeze and post-thaw product platelet content and recovery were collected to analyze the effects of dimethyl sulfoxide (DMSO) type, duration of frozen storage (DMSO-1 max 12 years and DMSO-2 frozen DTC max 4 years at -80°C) and type of plasma used to suspend DTC. Coagulation characteristics of thawed DTC, plasma and supernatant of DTC (2× 2500 G) were measured with Kaolin thromboelastography (TEG) and phospholipid (PPL) activity assay. Platelet content and recovery of DTC is ±10%-15% lower in short-stored products and remained stable when stored beyond 0.5 years. Thawed DTC (n = 1724) were compliant to the European guidelines (98.1% post-thaw product recovery ≥50% from original product, 98.3% ≥200 × 109 platelets/unit). Compared to DMSO-1, products frozen with DMSO-2 showed ±8% reduced thaw-freeze recovery, a higher TEG clot strength (MA 58 [6] vs. 64 [8] mm) and same ±11 s PPL clotting time. The use of cold-stored thawed plasma instead of fresh thawed plasma did not influence product recovery or TEG-MA. Regardless of alterations, product quality was in compliance with European guidelines and unaffected by storage duration up to 12 years of -80°C frozen storage.


Status dystonicus associated with CLN8 disease.
Status dystonicus is an underdiagnosed condition, probably due to heterogeneous etiology, presentation and course. Herein, we report the first case of CLN8 disease in the literature presenting with status dystonicus who responded well to pharmacological intervention. A boy aged five years and three months presented with fever, loss of appetite, intermittent excessive dystonic contractions, opisthotonus with retrocollis, and irritability for three days. His developmental milestones were reported as normal up to the age of three years and six months. At this age, he developed seizures, ataxia, and vision problems. Deterioration in developmental milestones was observed from the age of four. Laboratory tests demonstrated leukocytosis, abnormal renal function, mild metabolic acidosis, elevated creatine kinase and transaminase levels. The brain magnetic resonance imaging demonstrated cerebral and cerebellar atrophy. Homozygous missense mutation of c.709G > A (p.G237R) in the CLN8 gene was revealed. With all these clinical and laboratory findings, he was diagnosed with status dystonicus associated with CLN8 disease. Antibiotherapy, anticonvulsant drugs, and intravenous hydration with alkaline fluids were initiated. Due to irregular breathing, dysphagia, and worsening of dystonic contractions, mechanical ventilation was performed, and baclofen, haloperidol, midazolam infusion and chloral hydrate were administered, respectively. Finally, serum creatine kinase levels decreased, and dystonic contractions improved on the 15th day of hospitalization. To the best of our knowledge, our case is the first report describing the status dystonicus in a patient with CLN8 disease. Our report suggested that neuronal ceroid lipofuscinoses should be kept in mind in the etiology of status dystonicus.


Expression of aromatase mRNA and effects of aromatase inhibitor during ovarian development in the medaka, Oryzias latipes.
In teleost fish, several studies have implicated estrogens in the process of ovarian development, but the exact role of endogenous estrogen is still unclear. We examined the expression of aromatase mRNA with in situ hybridization, and the effects of Fadrozole, a nonsteroidal aromatase inhibitor (AI), during ovarian development in medaka Oryzias latipes. Medaka aromatase was first detected on the ventral side of ovaries from four to 10 days after hatching (dah), after occurrence of oogenesis. AI treatment after hatching suppressed the ovarian cavity formation from 30 dah but did not affect early oogenesis and folliculogenesis during ovarian development. These results suggest that endogenous estrogen is specifically required for formation of the ovarian cavity, but is not essential for early oogenesis and folliculogenesis in medaka.


Sleep is not just for the brain: transcriptional responses to sleep in peripheral tissues.
Many have assumed that the primary function of sleep is for the brain. We evaluated the molecular consequences of sleep and sleep deprivation outside the brain, in heart and lung. Using microarrays we compared gene expression in tissue from sleeping and sleep deprived mice euthanized at the same diurnal times. In each tissue, nearly two thousand genes demonstrated statistically significant differential expression as a function of sleep/wake behavioral state. To mitigate the influence of an artificial deprivation protocol, we identified a subset of these transcripts as specifically sleep-enhanced or sleep-repressed by requiring that their expression also change over the course of unperturbed sleep. 3% and 6% of the assayed transcripts showed "sleep specific" changes in the lung and heart respectively. Sleep specific transcripts in these tissues demonstrated highly significant overlap and shared temporal dynamics. Markers of cellular stress and the unfolded protein response were reduced during sleep in both tissues. These results mirror previous findings in brain. Sleep-enhanced pathways reflected the unique metabolic functions of each tissue. Transcripts related to carbohydrate and sulfur metabolic processes were enhanced by sleep in the lung, and collectively favor buffering from oxidative stress. DNA repair and protein metabolism annotations were significantly enriched among the sleep-enhanced transcripts in the heart. Our results also suggest that sleep may provide a Zeitgeber, or synchronizing cue, in the lung as a large cluster of transcripts demonstrated systematic changes in inter-animal variability as a function of both sleep duration and circadian time. Our data support the notion that the molecular consequences of sleep/wake behavioral state extend beyond the brain to include peripheral tissues. Sleep state induces a highly overlapping response in both heart and lung. We conclude that sleep enhances organ specific molecular functions and that it has a ubiquitous role in reducing cellular metabolic stress in both brain and peripheral tissues. Finally, our data suggest a novel role for sleep in synchronizing transcription in peripheral tissues.


Predictors of walking performance and walking capacity in people with lumbar spinal stenosis, low back pain, and asymptomatic controls.
To examine predictors of community walking performance and walking capacity in people with lumbar spinal stenosis (LSS), compared with people with low back pain and asymptomatic control subjects. Retrospective analysis. University spine program. Participants (N=126; 50 LSS, 44 low back pain, 32 asymptomatic control subjects) aged 55 to 80 years were studied. Not applicable. Seven-day community walking distance measured by pedometer (walking performance) and a 15-minute walking test (walking capacity). All participants had lumbosacral magnetic resonance imaging, electrodiagnostic testing, and a history and physical examination, including a history of pain and neurologic symptoms, a straight leg raise test, and tests for directional symptoms, reflexes, strength, and nerve tension signs. The study questionnaire included demographic information, a history of back/leg pain, and questions about walking, exercise frequency, and pain level, as well as the standardized Quebec Back Pain Disability Scale. Body mass index (BMI), pain, age, and female sex predicted walking performance (r(2)=.41) and walking capacity (r(2)=.41). The diagnosis of LSS itself had no clear relationship with either walking variable. Compared with the asymptomatic group, LSS participants had significantly lower values for all walking parameters, with the exception of stride length, while there was no significant difference between the LSS and low back pain groups. BMI, pain, female sex, and age predict walking performance and capacity in people with LSS, those with low back pain, and asymptomatic control subjects. While pain was the strongest predictor of walking capacity, BMI was the strongest predictor of walking performance. Average pain, rather than leg pain, was predictive of walking performance and capacity. Obesity and pain are modifiable predictors of walking deficits that could be targets for future intervention studies aimed at increasing walking performance and capacity in both the low back pain and LSS populations.


Carvacrol reduces blood pressure, arterial responsiveness and increases expression of MAS receptors in spontaneously hypertensive rats.
To analyze the effect of the use of carvacrol in the cardiovascular system of spontaneously hypertensive rats (SHR). Methods: Twenty animals were allocated in four groups, one group control Wistar receiving only sorbitol, used as vehicle of administration of the carvacrol (Wistar-Vehicle), one control group SHR, also receive only sorbitol (SHR-Vehicle), a third, treated with losartan (SHR-Losartan/50 mg/kg), and the fourth, treated with carvacrol (SHR - Carvacrol/20 mg/kg). Sorbitol, losartan and carvacrol were administered by oral gavage daily for 30-day. Hemodynamic variables, vascular reactivity, biochemical parameters, and expression of Mas and AT1 receptors in kidney tissues were analyzed. SHR- Carvacrol group showed a maximal effect of inhibition of 56% in the curve of norepinephrine. The Emax of the curves with Ca2+ were smaller in the groups SHR-losartan (40.17%) and SHR-carvacrol (35.71%) when compared to the SHR-Vehicle. The carvacrol increased the expression of the MAS receptors in kidney tissue. Thirty days of treatment with carvacrol showed an antihypertensive effect associated with less peripheral vascular resistance. Also, treatment with carvacrol increased the expression of MAS receptors in kidney tissue.


Concise review: Human cell engineering: cellular reprogramming and genome editing.
Cell engineering is defined here as the collective ability to both reset and edit the genome of a mammalian cell. Until recently, this had been extremely challenging to achieve as nontransformed human cells are significantly refractory to both these processes. The recent success in reprogramming somatic cells into induced pluripotent stem cells that are self-renewable in culture, coupled with our increasing ability to effect precise and predesigned genomic editing, now readily permits cellular changes at both the genetic and epigenetic levels. These dual capabilities also make possible the generation of genetically matched, disease-free stem cells from patients for regenerative medicine. The objective of this review is to summarize the key enabling developments on these two rapidly evolving research fronts in human cell engineering, highlight unresolved issues, and outline potential future research directions.


Contraction stress testing for primary foetal surveillance by stimulated endogenous oxytocin release.
A test for uteroplacental insufficiency based on the idea that a given maternal-foetal unit could be tested by decreasing intervillous space blood flow, offers an individual test for each foetus. Clinically, the use of uterine activity to decrease intervillous space blood flow is the most practical. Following Freeman's suggestion, authors use an endogenous oxytocin test, where the oxytocin discharge is induced by nipple stimulation (EOS).


Alpha-sialyl cholesterol increases laminin in Schwann cell cultures and attenuates cytostatic drug-induced reduction of laminin.
Schwann cells play an important role in peripheral nerve regeneration. Here, we report the effect of alpha-sialyl cholesterol (alpha-SC), a derivative of the sialic acid-containing natural gangliosides, and the cytostatic agents, cisplatin, taxol and vincristine on the laminin production in Schwann cell cultures isolated from rat sciatic nerves. Laminin, one of several extracellular matrix components produced by Schwann cells, is known to potentiate axonal outgrowth. Laminin content was increased by alpha-SC, starting at 7.0 micrograms/ml with a maximal effect at 22.4 micrograms/ml (30%, P < 0.001). The three cytostatic drugs, dose-dependently reduced laminin content in Schwann cell cultures: (1) cisplatin at a threshold dose of 2 micrograms/ml (-26.4%, P < 0.001); (2) taxol, starting at a dose of 1 ng/ml (-8.0%, P < 0.05); and (3) vincristine, starting at 0.5 ng/ml (-5.9%, P < 0.05). Cultured Schwann cells were incubated with cytostatic drugs in combination with increasing amounts of alpha-SC and it was found that, depending on the cytostatic drug concentration used, alpha-SC could reduce or completely prevent the cytostatic drug-induced reduction of laminin in Schwann cell cultures. Co-treatment with alpha-SC also reduced part of the morphological changes caused by the cytostatic drugs. alpha-SC did not counteract the anti-proliferative effect of the cytostatic drugs on K-562 human erythroleukemia cells. In conclusion, alpha-SC increased laminin content in Schwann cell cultures and protected Schwann cell cultures against the decrease of laminin by cytostatic drugs without interfering with the anti-proliferative potential, suggesting that alpha-SC may have clinical use in protecting cancer patients against the neurotoxic effects of cytostatic drugs.


[The effect of a single firing stimulation of the vagus nerve on atrioventricular behavior of stimulation in the cat heart].
Dromotropic vagal effect was shown to consist of a prolonged tonic component and a short-term one depending on the interval between the burst irritation of the vagus nerve and the P-wave of the ECG.


Incidence of second primary cancers among survivors of childhood cancer: A population-based study, Osaka, Japan, 1975-2015.
Second primary cancer (SPC) is one of the most life-threatening late effects of childhood cancers. We investigated the incidence and survival outcomes of SPC in childhood cancer patients in Japan. Data were obtained from the population-based Osaka Cancer Registry. Individuals diagnosed with cancer at age 0-14 years during 1975-2014 and survived 2 months or longer were followed through December 2015. The risk of developing SPC was assessed with standardized incidence ratio (SIR), excess absolute risk (EAR, per 100,000 person-years), and cumulative incidence. Multivariable Poisson regression analysis was carried out to assess relative risks of SPC by treatment method. Survival analysis was undertaken using the Kaplan-Meier method. Of 7229 childhood cancer survivors, 101 (1.4%) developed SPC after a median of 11.6 years. Overall SIR was 5.0, which corresponded with 84.3 EAR. The cumulative incidence was 0.9%, 2.1%, and 3.4% at 10, 20, and 30 years, respectively. Among all SPCs, the type that contributed most to the overall burden was cancers in the central nervous system (EAR = 28.0) followed by digestive system (EAR = 15.1), thyroid (EAR = 8.3), and bones and joints (EAR = 7.8); median latency ranged from 2.0 years (lymphomas) to 26.6 years (skin cancers). Patients treated with radiotherapy alone were at a 2.58-fold increased risk of developing SPC compared to those who received neither chemotherapy nor radiotherapy. Among patients who developed SPCs, 5-year and 10-year survival probabilities after SPC diagnosis were 61.7% and 52.0%, respectively. Risk-based long-term follow-up planning is essential to inform survivorship care and help reduce the burden of SPCs in childhood cancer survivors.


Performance of serum apolipoprotein-A1 as a sentinel of Covid-19.
Since 1920, a decrease in serum cholesterol has been identified as a marker of severe pneumonia. We have assessed the performance of serum apolipoprotein-A1, the main transporter of HDL-cholesterol, to identify the early spread of coronavirus disease 2019 (Covid-19) in the general population and its diagnostic performance for the Covid-19. We compared the daily mean serum apolipoprotein-A1 during the first 34 weeks of 2020 in a population that is routinely followed for a risk of liver fibrosis risk in the USA (212,297 serum) and in France (20,652 serum) in relation to a local increase in confirmed cases, and in comparison to the same period in 2019 (266,976 and 28,452 serum, respectively). We prospectively assessed the sensitivity of this marker in an observational study of 136 consecutive hospitalized cases and retrospectively evaluated its specificity in 7,481 controls representing the general population. The mean serum apolipoprotein-A1 levels in the survey populations began decreasing in January 2020, compared to the same period in 2019. This decrease was highly correlated with the daily increase in confirmed Covid-19 cases in the following 34 weeks, both in France and USA, including the June and mid-July recovery periods in France. Apolipoprotein-A1 at the 1.25 g/L cutoff had a sensitivity of 90.6% (95%CI84.2-95.1) and a specificity of 96.1% (95.7-96.6%) for the diagnosis of Covid-19. The area under the characteristics curve was 0.978 (0.957-0.988), and outperformed haptoglobin and liver function tests. The adjusted risk ratio of apolipoprotein-A1 for survival without transfer to intensive care unit was 5.61 (95%CI 1.02-31.0; P = 0.04). Apolipoprotein-A1 could be a sentinel of the pandemic in existing routine surveillance of the general population. NCT01927133, CER-2020-14.


Frequency and implication of focal fatty sparing in segmental homogeneous fatty liver at ultrasound.
The purpose of this study was to investigate whether focal fatty sparing arises from preexisting segmental homogeneous nonalcoholic fatty liver (NAFL). Patients with various patterns of fatty liver were recruited from among 16,127 people, and 132 participants with segmental homogeneous NAFL were followed during the second year. All participants underwent ultrasonography. The sonographic findings in segmental homogeneous NAFL were analyzed. Of the 16,127 participants, 1,619 had fatty liver (10.0 %), and of these 1,568 had NAFL (9.72 %) and 51 had alcoholic fatty liver (0.31 %). Of the 1,568 patients with NAFL, 176 had homogeneous fatty liver with focal fatty sparing (11.2 %), 132 had segmental homogeneous fatty liver (8.42 %), and the rest had other patterns. At follow-up, of the 132 patients with segmental homogeneous fatty liver, 3 (2.3 %) were found to have a normal liver and 26 (19.7 %) diffuse homogeneous and heterogeneous fatty liver, and no focal fatty sparing was found (P < 0.001). Focal fatty sparing usually does not arise in preexisting segmental homogeneous NAFL. This implies that newly appearing nodular hypoechogenicity in segmental homogeneous fatty liver is more likely to be a true lesion rather than focal fatty sparing.


Rescue from Sexually Dimorphic Neuronal Cell Death by Estradiol and PI3 Kinase Activity.
Responses of primary hippocampal and cortical neurons derived from male and female rats to cellular stressors were studied. It is demonstrated that 17β-estradiol (E2), a potent neuroprotectant, protected the female neurons but had no effects on the male neurons from CoCl2- and glutamate-induced toxicity. Agonists of the estrogen receptor (ER) subtypes ERα and ERβ, DPN and PPT, respectively, had similar effects to E2. By contrast, effects of E2 were abolished by the ER antagonist ICI-182780, further corroborating the neuroprotective role of ERs. In male neurons, CoCl2 predominately activated the apoptosis-inducing factor (AIF)-dependent pathway and AIF translocation from the cytosol to the nucleus. In comparison, CoCl2 activated the caspase pathway and cytochrome c release in female neurons. The inhibitors of these pathways, namely DiQ for AIF and zVAD for caspase, specifically rescued CoCl2-induced cell death in male and female neurons, respectively. When zVAD and ICI-182780, and E2 were applied in combination, it was demonstrated E2 acted on the caspase pathway leading to female-specific neuroprotection. Furthermore, the PI3 kinase (PI3K) inhibitor blocked the rescue effects of DiQ and zVAD on the male and female neurons, respectively, suggesting that PI3K is a common upstream regulator for both pathways. The present study suggested that both sex-specific and nonspecific mechanisms played a role in neuronal responses to stressors and protective reagents.


Sharing individualised template MRI data for MEG source reconstruction: A solution for open data while keeping subject confidentiality.
The increasing requirements for adoption of FAIR data management and sharing original research data from neuroimaging studies can be at odds with protecting the anonymity of the research participants due to the person-identifiable anatomical features in the data. We propose a solution to this dilemma for anatomical MRIs used in MEG source analysis. In MEG analysis, the channel-level data is reconstructed to the source-level using models derived from anatomical MRIs. Sharing data, therefore, requires sharing the anatomical MRI to replicate the analysis. The suggested solution is to replace the individual anatomical MRIs with individualised warped templates that can be used to carry out the MEG source analysis and that provide sufficient geometrical similarity to the original participants' MRIs. First, we demonstrate how the individualised template warping can be implemented with one of the leading open-source neuroimaging analysis toolboxes. Second, we compare results from four different MEG source reconstruction methods performed with an individualised warped template to those using the participant's original MRI. While the source reconstruction results are not numerically identical, there is a high similarity between the results for single dipole fits, dynamic imaging of coherent sources beamforming, and atlas-based virtual channel beamforming. There is a moderate similarity between minimum-norm estimates, as anticipated due to this method being anatomically constrained and dependent on the exact morphological features of the cortical sheet. We also compared the morphological features of the warped template to those of the original MRI. These showed a high similarity in grey matter volume and surface area, but a low similarity in the average cortical thickness and the mean folding index within cortical parcels. Taken together, this demonstrates that the results obtained by MEG source reconstruction can be preserved with the warped templates, whereas the anatomical and morphological fingerprint is sufficiently altered to protect the anonymity of research participants. In cases where participants consent to sharing anatomical MRI data, it remains preferable to share the original defaced data with an appropriate data use agreement. In cases where participants did not consent to share their MRIs, the individualised warped MRI template offers a good compromise in sharing data for reuse while retaining anonymity for research participants.


Rare alkaline phosphatase-harboring bacteria mediate organic phosphorus mineralization during swine manure composting.
Deciphering ecological functions of alkaline phosphatase (phoD)-harboring bacteria in composting systems is crucial but poorly understood. High-throughput sequencing, gene quantification, and statistical analyses were applied to investigate effects of abundance and diversity of phoD-harboring bacteria (PHB) on phosphorus availability during swine manure composting. Results showed that available phosphorus notably increased from 0.5 to 1.43 g kg-1, and physicochemical properties and enzyme activities affected PHB community composition. Phylogenetic signals of PHB responded notably to temperature and phosphorus components, and stochasticity (94.2 %) dominated community assembly. Abundance and diversity of PHB directly and indirectly influenced phosphorus availability, and rare PHB mediated organic phosphorus mineralization. A phosphate-solubilizing bacterium (PSB) Pseudomonas sp. WWJ-22 isolated from compost displayed good efficiency in mineralizing lecithin, demonstrating the highest phosphorus-solubilizing level of 116.3 mg L-1. This study highlights ecological roles of PHB on phosphorus availability and provides a potential PSB candidate for composting.


Women's Empowerment and Infant and Child Health Status in Sub-Saharan Africa: A Systematic Review.
Although many studies have examined the relationship between women's empowerment and a wide range of health outcomes, the extent to which the different dimensions of empowerment influence children's health, and through which mechanisms and in what contexts, is limited in sub-Saharan Africa. The objective of this review is to systematically assess and examine studies that investigated the association between women's empowerment and children's health status in sub-Saharan Africa. A systematic review of the published literature is searched through PubMed, Google Scholar, Embase, Web of Science and Scopus databases focusing on different measures of women's empowerment and children's health outcomes. Inclusion criteria in the review are studies that are published in English; full and original articles; studies measuring at least one dimension of women's empowerment and children's health outcomes; and Sub-Saharan African context. Studies included in this review are articles published between the year 2000 and 2019. Studies were excluded if the source was a letter, editorial, review, commentary, abstracts without providing full information about the study. Initially 4718 citations were identified. Finally, 15 studies met the inclusion and exclusion criteria. In general, the evidence suggests that women's empowerment at the household level is positively and statistically significantly associated with better children's health outcomes in sub-Saharan African countries. The review also reveals that women's decision-making power or autonomy is the most common measure of women's empowerment employed by many studies. Future related studies would benefit by incorporating additional aspects of women's empowerment and child health outcomes.


[Organization of services for treatment of postoperative pain--3-year experience].
We report organisation principles and three year experience of Acute Pain Service in general surgery clinic. 481 patients were treated after abdominal and vascular interventions, hemorrhoidal varices and mammectomies. Continuous epidural, combined spinal-epidural, intrapleural anaesthesia and continuous brachial plexus block were used for pain control. Time of analgesia varied from 1 to 4 days. The level of analgesia was assessed as good (VAS 3) in 94.8% of cases. Complications were mainly technical due to catheter or antibacterial filter failure. In 2% of cases cardiovascular complications were observed. Respiratory depression occurred in 1 patient. The work of APS team was assessed as very good by both surgeons and patients.


Neurotransmitters alter the numbers of synapses and organelles in photoreceptor terminals in the lamina of the housefly, Musca domestica.
Various organelles in the lamina terminals of housefly photoreceptors exhibit daily rhythms having a circadian basis. These include changes in the numbers of photoreceptor tetrad and L2 feedback synapses, and longitudinal movements of screening pigment. Circadian information has previously been suggested to spread from the clock to the lamina via widefield cells expressing either 5-hydroxytryptamine or pigment-dispersing hormone-like immunoreactivity. We examined the action of these neuromodulators, and other candidate neurotransmitters, 4 h after injecting either the transmitter or a control into the medulla. We counted electron microscope profiles of organelles that normally exhibit circadian changes, and two types of invagination into photoreceptor terminals, capitate projections and inter-receptor invaginations. No single substance mediated the changes observed. Injected pigment-dispersing hormone peptide decreased the number of pigment granules, implicating this peptide in screening pigment migration, but produced no changes in synapse-related organelles. alpha-Aminobutyric acid exclusively decreased the number of L2 feedback synapses. Responses to other transmitters were specific, and often large, but generally not statistically significant. Histamine, for example, may decrease the number of tetrads, possibly by direct autoregulation. The results suggest that there is likely to be more than one effector in the circadian pathways to the lamina.


Proconvulsant effects of aminophylline during amygdala kindling in developing rats.
Aminophylline (A) is a proconvulsant in adult rats. We studied the effect of A on amygdala kindling in 15-day-old rat pups. Production of generalized seizures was significantly promoted by A at doses ranging from 10 to 100 mg/kg. Terminal status epilepticus (TSE) was produced in 33% of pups receiving 25 mg/kg A, 75% of pups receiving 50 mg/kg A, and 100% of pups receiving 100 mg/kg A. The number of stimuli needed to produce a stage 4-5 generalized seizure was significantly smaller in animals receiving 10 mg/kg A (5.7 +/- 3.4), 25 mg/kg A (3.4 +/- 2.4), 50 mg/kg A (1.9 +/- 1.4), or 100 mg/kg A (1.9 +/- 1.6) than in saline-treated controls (12.3 +/- 3.7) (P less than 0.001). In addition, 16% of pups receiving 50 mg/kg and 33% of pups receiving 100 mg/kg A and never stimulated developed TSE. These seizure-promoting effects of A in rat pups undergoing amygdala kindling are far more dramatic and occur at far lower doses than those previously reported in adults.


Extraoral digestion: outsourcing the role of the hemipteran midgut.
Extraoral digestion allows for breakdown of dietary components before they reach the midgut for final enzymatic degradation and absorption. In the Hemiptera, this is achieved by the secretion of enzyme-rich fluids from the salivary gland, with the combination of protein and mRNA from these tissues termed the sialome. Separate channels within the hemipteran stylets allow for secretion of saliva and ingestion of predigested material in a non-reflux mechanism. Both feeding mode and diet type influence the composition of the hemipteran sialome, as illustrated by 1) differences in protease abundance between hematophagous and predatory heteropteran sialomes, 2) diet specific aminopeptidase-N genes among aphid biotypes, and 3) adaptation-induced sialome variation in related cicada populations. Despite challenges associated with incomplete genome annotation, -omics analysis of the sialomes of diverse hemipteran species will enhance understanding of both sialome function and the evolution of extraoral digestion within the order.


Metalloproteinase with factor X activating and fibrinogenolytic activities from Vipera berus berus venom.
We have previously shown that Vipera berus berus venom contains several factor X activating enzymes. In the present study we have investigated one of them. The enzyme was separated from venom by gel filtration on Sephadex G-100 superfine and chromatography on agarose HPS-7 and phenyl-agarose. The enzyme is a glycosylated metalloproteinase containing hexoses, hexosamines and neuraminic acid. The purified factor X activating enzyme consists of two equal chains (59 kDa). The specificity studies have shown that enzyme is nonspecific factor X activating proteinase hydrolysing also proteins such as azocasein, gelatin and fibrinogen. The enzyme hydrolyses oxidized insulin B-chain at the positions Ala(14)-Leu(15) and Tyr(16)-Leu(17) but it is inactive on fibrin, plasminogen and prothrombin. We used 8-10 amino acid residues containing peptides, which reproduce the sequence around the cleavage sites in factor X, factor IX and fibrinogen, as potential substrates for enzyme. Cleavage products of peptide hydrolysis were determined by MALDI-TOF MS. The peptide Asn-Asn-Leu-Thr-Arg-Ile-Val-Gly-Gly-factor X fragment was cleaved by enzyme at positions Leu(3)-Thr(4) and Arg(5)-Ile(6). The fibrinogen peptide fragment Glu-Tyr-His-Thr-Glu-Lys-Leu-Val-Thr-Ser was hydrolysed at position Lys(6)-Leu(7).


Identification of Fusarium graminearum in cereal samples by DNA Detection Test Strips.
Development of a fast, sensitive and easy to handle method for the detection of Fusarium graminearum contamination in cereal samples by PCR. DNA Detection Test Strips were used for PCR-product detection and the method was compared to agarose gel electrophoresis. A minimum of 0.26 ng of purified target DNA was detectable with the Test Strip Detection limit in less contaminated samples was slightly lower when gel electrophoresis was used for amplicon detection. In highly contaminated samples, detection limits of both methods were similar. Detection of PCR products was performed in 20 min without the need of special technical equipment or hazardous fluorescent DNA dyes. The new method described is useful for the screening of cereals in industrial quality control.


An evolutionarily conserved charged residue dictates the specificity of heterophilic interactions among nectins.
Nectins are a family of four cell surface glycoproteins belonging to the immunoglobulin superfamily that mediate cell-cell adhesion and associated signalling pathways, thereby regulating several physiological processes including morphogenesis, growth and development of multicellular organisms. Nectins interact among themselves through their extracellular domains from the adjacent cells in both homophilic and heterophilic fashions to support cell-cell adhesion. Although nectins form homodimers as demonstrated in experimental set-ups, only the specific heterophilic interactions among nectins are physiologically relevant as shown by in vivo studies. It has been hypothesised that a conserved charged residue present at the binding interface acts as the molecular switch for heterophilic nectin-nectin recognitions. In this work, we have analysed the energetics of homophilic and heterophilic interactions of nectins, followed by surface plasmon resonance-based binding studies and complementary in silico analyses. Our findings confirm that the conserved charged residues at the binding interfaces dictate the specificity of the nectin-nectin heterophilic interactions. Furthermore, these residues also play a role in conferring higher affinity to the heterophilic interactions, thereby making them physiologically more prevalent compared to homophilic interactions. Thus, this work reveals the molecular basis of heterophilic recognitions among nectins that contribute to their physiological functions.


Dynamic pattern of estradiol binding to uterine receptors of the rat. Inhibition and stimulation by unsaturated fatty acids.
The binding of estradiol to uterine cytosoluble receptors from 24-day-old rats was reduced or potentiated by unsaturated fatty acids (NEFAs), depending on the concentrations of estradiol and unsaturated NEFAs. At estradiol concentrations of up to 1.5 x 10(-8) M, unsaturated NEFAs inhibited estradiol binding to the 8 S cytosol receptor. This inhibition was dose-dependent (10-70%, p less than 0.001) and a function of NEFA unsaturation. Scatchard analysis indicated that unsaturated NEFAs caused a large decrease in receptor affinity for estradiol. Polyunsaturated NEFAs had no apparent effect on estradiol binding at estradiol concentrations of 2-4 x 10(-8) M. At high estradiol concentrations (above 4 x 10(-8) M), estradiol binding was increased 130-250% (p less than 0.01) by polyunsaturated NEFAs. This increased binding was particularly associated with proteins sedimenting at 12.5 S and the 8 S binding was, in fact, reduced. Metabolic studies showed that the reduced binding in the presence of unsaturated fatty acids was correlated with a decrease in reversibly bound estradiol at low estradiol concentrations. The increase in estradiol binding at high estradiol concentrations is the result of a reduction in reversibly bound estradiol and an increase in nonorganic solvent-extractable (water-soluble) estradiol. The amounts of these water-soluble estradiol derivatives depended on both estradiol and unsaturated NEFA concentrations. 70% of the water-soluble estradiol derivatives were trichloroacetic acid-precipitable, suggesting a covalent protein-steroid link. Thus, changes in the hydrophobic fatty acid environment of the uterine cytosol estrogen receptor could modify estrogen-receptor function by altering binding site conformation and/or by inducing changes in estradiol metabolism.


Combination of antibiotics suppressed the increase of a part of ARGs in fecal microorganism of weaned pigs.
The presence of antibiotic resistance genes (ARGs) is one of the most important public health concerns. Six tetracycline resistance genes (TRGs-tetA, tetC, tetL, tetO, tetW, and tetX) were quantified using real-time quantitative polymerase chain reaction (qPCR) in the fecal microorganisms of weaned pigs. Two hundred 35-day-old weaned pigs were fed different dietary antibiotics for 28 days: (1) no antibiotic as the control treatment (CT); (2) chlortetracycline, bacitracin zinc and colistin sulfate (CBC); (3) bacitracin zinc and colistin sulfate (BC); and (4) chlortetracycline (CTC). The detection frequencies (DFs) of tetC, tetL, and tetW were 100 %; and the DFs of tetA, tetD, tetM, tetO, and tetX were 65 %. The relative abundances (tet/16S rRNA gene copy numbers) of six tet genes (tetA, tetC, tetL, tetO, tetW and tetX) were between 1.5 × 10(-4) and 2.0 × 10(-1). In the group CTC, the relative abundances of tetC (P < 0.01), tetL (P < 0.01), tetO (P < 0.05), tetW (P < 0.01), and tetX (P < 0.01) were greater than those of the group CT. Compared with the group CTC, the relative abundances of tetC (P < 0.01), tetL (P < 0.01), and tetW (P < 0.01) were decreased in the CBC and BC groups. These results indicate that a combination of different antibiotics suppressed the abundance increase of a part of tet genes, which suggests that a combination of antibiotics produces multiple selection pressures on fecal microorganism of pigs.


Effects of biochar and biofertilizer on cadmium-contaminated cotton growth and the antioxidative defense system.
Consistent use of large amounts of fertilizers, pesticides, and mulch can cause the accumulation of harmful substances in cotton plants. Among these harmful substances, cadmium (Cd), an undegradable element, stands out as being particularly highly toxic to plants. The objective of this study was to evaluate the ability of biochar (3%) and biofertilizer (1.5%) to decrease Cd uptake, increase cotton dry weight, and modulate the activities of photosynthetic and peroxidase (POD), superoxide dismutase (SOD), catalase enzyme (CAT) in cotton (Gossypium hirsutum L.) grown in Cd-contaminated soil (0, 1, 2, or 4 mg Cd kg-1 soil) in pots. These studies showed that, as expected, exogenous Cd adversely affects cotton chlorophyll and photosynthesis. However, biochar and biofertilizer increased cotton dry weight by an average of 16.82% and 32.62%, respectively. Meanwhile, biochar and biofertilizer decreased the accumulation of Cd in cotton organs, and there was a significant reduction in the amount of Cd in bolls (P < 0.05). Biochar and biofertilizer have a positive impact on cotton chlorophyll content, net photosynthesis, stomatal conductance, transpiration rate, and intercellular CO2 concentration. Thus, the addition of biochar and biofertilizer promote cotton growth. However, biochar and biofertilizer increased the SOD activity of leaves (47.70% and 77.21%), CAT activity of leaves (35.40% and 72.82%), SOD activity of roots (33.62% and 39.37%), and CAT activity of roots (36.91% and 60.29%), respectively, and the addition of biochar and biofertilizer decreased the content of MDA and electrolyte leakage rate. Redundancy analyses showed that biochar and biofertilizer also improved SOD and POD activities by reducing the heavy metal-induced oxidative stress in cotton and reducing Cd uptake in cotton organs. Therefore, biochar and biofertilizer have a positive effect on the growth of cotton.


Regulation of heterologous subtilin production in Bacillus subtilis W168.
Subtilin is a peptide antibiotic (lantibiotic) natively produced by Bacillus subtilis ATCC6633. It is encoded in a gene cluster spaBTCSIFEGRK (spa-locus) consisting of four transcriptional units: spaS (subtilin pre-peptide), spaBTC (modification and export), spaIFEG (immunity) and spaRK (regulation). Despite the pioneer understanding on subtilin biosynthesis, a robust platform to facilitate subtilin research and improve subtilin production is still a poorly explored spot. In this work, the intact spa-locus was successfully integrated into the chromosome of Bacillus subtilis W168, which is the by far best-characterized Gram-positive model organism with powerful genetics and many advantages in industrial use. Through systematic analysis of spa-promoter activities in B. subtilis W168 wild type and mutant strains, our work demonstrates that subtilin is basally expressed in B. subtilis W168, and the transition state regulator AbrB strongly represses subtilin biosynthesis in a growth phase-dependent manner. The deletion of AbrB remarkably enhanced subtilin gene expression, resulting in comparable yield of bioactive subtilin production as for B. subtilis ATCC6633. However, while in B. subtilis ATCC6633 AbrB regulates subtilin gene expression via SigH, which in turn activates spaRK, AbrB of B. subtilis W168 controls subtilin gene expression in SigH-independent manner, except for the regulation of spaBTC. Furthermore, the work shows that subtilin biosynthesis in B. subtilis W168 is regulated by the two-component regulatory system SpaRK and strictly relies on subtilin itself as inducer to fulfill the autoregulatory circuit. In addition, by incorporating the subtilin-producing system (spa-locus) and subtilin-reporting system (PpsdA-lux) together, we developed "online" reporter strains to efficiently monitor the dynamics of subtilin biosynthesis. Within this study, the model organism B. subtilis W168 was successfully established as a novel platform for subtilin biosynthesis and the underlying regulatory mechanism was comprehensively characterized. This work will not only facilitate genetic (engineering) studies on subtilin, but also pave the way for its industrial production. More broadly, this work will shed new light on the heterologous production of other lantibiotics.