Tumor suppressor genes studied by cell hybridization and chromosome transfer.
Although the cloning of tumor suppressor genes has proved to be an arduous task, often involving several years of labor intensive cloning strategies, a greater understanding of neoplastic progression will be made once the function and role of these genes have been sorted out. To fully appreciate the state at which this field of research currently is, however, one must understand that the road to tumor suppression was paved by both somatic cell hybridization and chromosome transfer studies. Although regarded by many as laboratory artifact, somatic cell hybridization has provided strong circumstantial evidence, if not formal proof, for the existence of tumor suppressor genes. In further reducing the complexity associated with whole genome transfer, single chromosome transfer was subsequently developed as a refinement to this technique so that one could unequivocally correlate suppression with a particular chromosome. We have learned from these studies that single chromosomes harbor the genetic information necessary to reverse the malignant phenotype associated with cancer cells. Furthermore, multiple tumor suppressor loci are now known to exist, with one to several different loci associated with a given tumor type. In this review, we present evidence for tumor suppressor genes and discuss the early studies that led to their discovery.


In vivo semen-associated pH neutralization of cervicovaginal secretions.
Physiological cervicovaginal acidity can partly inactivate human immunodeficiency virus (HIV). Basic semen components should be able to partially neutralize in vivo cervicovaginal pH. The goals of the study were to evaluate the relationship between cervicovaginal pH and presence of semen components in sexually active African women and to assess whether vaginal douching with water performed just after sexual intercourse could significantly reduce semen components and restore physiological cervicovaginal pH. Cervicovaginal secretion (CVS) from 56 heterosexual African women (19 to 45 years old), living in Bangui, Central African Republic, were evaluated for pH, semen components (prostatic acid phosphatase [PAP] and prostatic specific antigen [PSA]), cellularity, and hemoglobin at inclusion and after vaginal douching with 100 ml of water by using a bock. Before douching, semen components were found in 46 of 56 CVS (82%). The mean vaginal pH was 5.2 (range, 3.6 to 7.7), and concentrations of both PAP and PSA correlated positively and strongly with cervicovaginal pH (P < 0.001). After douching, semen components were found in 35 of 56 CVS (62%) (P = 0.03). Cervicovaginal PAP and PSA levels were significantly decreased (respectively, P < 0.0001 and P < 0.01; PAP, -72%; PSA, -87%), as was the total cell count (-60%; P < 0.0001). Furthermore, in CVS previously positive for both PAP and PSA, the mean vaginal pH was significantly decreased (6.5 versus 5.3, P < 0.01); no genital bleeding was observed. Frequent persistence of semen in CVS from heterosexually active African women leads to a shift from acidity to neutrality that could favor male to female HIV transmission. Vaginal douching provides significant elimination of semen after sexual intercourse; it should be considered for study as a supplementary means for the prevention of heterosexual HIV transmission.


Control of cyclic adenosine 3',5'-monophosphate production in osteoclasts: calcitonin-induced persistent activation and homologous desensitization of adenylate cyclase.
Hormonal control of cAMP production in the osteoclast has not been investigated in detail because this bone-resorbing cell has been difficult to isolate. We have used osteoclasts freshly isolated by disaggregation from neonatal rat long bones and settled onto coverslips (100-150 cells per coverslip) to examine the effects of calcitonin and prostaglandin E2 on osteoclast cAMP levels and cytoplasmic spreading. Salmon, eel, and human calcitonin (CT), and various analogs, stimulated cAMP production in a dose-dependent manner with relative potencies as seen in other response systems. Forskolin (10(-7) M) increased the sensitivity and amplitude of the response. Pretreatment with pertussis toxin (200 ng/ml for 3 h) had no effect suggesting that CT does not act through Ni, the inhibitory guanine nucleotide regulatory unit of adenylate cyclase. CT treatment was associated with rapid and dose-dependent induction of a persistent activated state of adenylate cyclase and homologous desensitization, the former being a particular feature of CT action previously observed in nonosteoclastic cells. Quantitative histomorphometry demonstrated a sensitive, dose dependent, and prolonged (greater than 2 h) reduction in osteoclast plan area after exposure to salmon CT. Although prostaglandin E2 also stimulated cAMP production and resulted in cell contraction in osteoclasts this was not associated with persistent activation of adenylate cyclase nor with prolonged contraction. Persistent activation of adenylate cyclase may be an important mechanism in CT inhibition of osteoclast function.


The pregnant orthopaedic patient and medication concerns.
It is not every day that nurses practicing in an orthopaedic setting must care for a pregnant patient. However, when they do, there are medication concerns about two patients--the pregnant patient and the fetus. The pregnant patient will need relief from pain and discomfort at the same time that the fetus will need to be protected from medications that pose potential fetal risk. In this article, the risk to the fetus from selected medications is identified, and the effect that normal physiologic changes of pregnancy may have on absorption, distribution, and elimination of medications is discussed.


[Results of the Czech National Colorectal Cancer screening programme - faecal occult blood tests].
The nationwide Colorectal Cancer Screening Programme was introduced in the Czech Republic in 2000. The aim of this article is to describe the employment of faecal occult blood tests (FOBTs) by the Czech population within the screening programme, and to provide information on the latest results of the programme. Data on the development of the colorectal cancer (CRC) burden in the Czech population is obtained from the Czech National Cancer Registry, a database required by the Czech law that has been collecting comprehensive data on cancer patients since 1977. Data on FOBT employment can be obtained from health care payers, and was provided by the Czech National Reference Centre. Around 8,000 patients are diagnosed with colorectal cancer in the Czech Republic each year, and the number of CRC deaths is about 4,000. Despite the ongoing screening programme, significant improvements in the proportional representation of cancer stages (i.e., improvements in early detection of CRC cases) have yet to be seen. Although the number of FOBTs performed in the Czech Republic has significantly grown in the long term (which is accompanied by an increase in coverage by this screening test), the total coverage of the Czech population aged over 50 was only 25.5% in 2012. The Olomouc Region, the Zlin region, and the Usti nad Labem region had the highest coverage rates by CRC screening based on FOBT (over 28%), while the Capital of Prague had the lowest coverage rate (18%). Since 2008, FOBT positivity rates have seen a continuous and significant increase, reaching 6.9% in 2012. Between 13 to 14% of FOBTs in women are performed by practical gynaecologists. Despite a significant increase in the participation rate in recent years, which was partially improved by the involvement of practical gynaecologists, the programme unfortunately still covers only a quarter of the eligible population. Implementation of effective measures aimed at getting people interested in preventive examinations (including the recently introduced programme of personalized invitations) is therefore essential; otherwise, the screening programme will not be successful on the population level.


Hydrolytic degradation of poly(ethylene oxide)-block-polycaprolactone worm micelles.
Spherical micelles and nanoparticles made with degradable polymers have been of great interest for therapeutic application, but degradation-induced changes in a spherical morphology can be subtle and mechanism/kinetics appears poorly understood. Here, we report the first preparation of giant and flexible worm micelles self-assembled from degradable copolymer poly(ethylene oxide)-block-polycaprolactone. Such worm micelles spontaneously shorten to generate spherical micelles, triggered by polycaprolactone hydrolysis, with distinct mechanism and kinetics from that which occurs in bulk material.


Positive association to IgE levels and a physical map of the 13q14 atopy locus.
Linkage of atopy and associated traits to a locus on chromosome 13q14 has been identified by several studies in diverse populations. We have previously shown the putative atopy gene to be contained within an interval of approximately 5 Mb flanked by D13S328 and D13S1269 and centred on D13S273. We have now extended this work using a top-down approach to physical mapping. A YAC contig was constructed covering the D13S328 and D13S1269 interval. Thirty-one ESTs were mapped to the contig. We constructed a BAC and PAC contig flanking D13S273 by approximately 750 kb in either direction. The interval contained 27 of the 31 ESTS from the YAC contig. Seven previously unknown microsatellites were recovered and then typed in two subject panels. A positive association between the total serum Immunoglobulin E concentration and the novel USAT24G1 microsatellite was discovered (P(corrected)<0.005) and replicated in a second panel of families. The discovery of a region of positive association within the BAC/PAC contig will permit identification of the atopy gene from this locus.


Predicting mortality after kidney transplantation: a clinical tool.
An increasing number of patients referred for transplantation are older and have complex comorbidity affecting outcome. Patient counseling is often empiric and time consuming. For the physician there are few clinical tools available to help quantify survival chances after transplantation. We used registry data to develop a series of tables that could be used in the clinical setting to predict survival probability. Using data from the Canadian Organ Replacement Registry, we generated clinical survival tables using Cox's regression model. Model covariates included age, race, gender, treatment period, primary renal disease cause, donor source, months on dialysis and comorbidities. A total of 6324 patients were included, 22% had > or =1 comorbid condition at baseline. After adjustment for age, gender and cause of renal disease, increased comorbidity was strongly associated with reduced patient-survival (P < 0.05). Age and comorbidity specific clinical survival tables showing the expected 1-, 3- and 5-year patient survival probabilities were generated. Separate tables were created for diabetics, nondiabetics, living-donor organs and deceased-donor transplantation. Patient-specific survival data can be estimated from registry data. We suggest annual or biannual tables generated by national registries across Europe and N. America, may be useful to those physicians faced with counseling patients and families.


Infective endocarditis: changing aetiology of disease.
Infective endocarditis (IE) is an evolving disease resulting in high morbidity and mortality. Despite medical and diagnostic advances, the incidence of the disease has remained unchanged, reflecting the changing epidemiological and microbiological profile of IE. Classical risk factors such as rheumatic heart disease have now been overtaken by new risk factors including an ageing population, degenerative valve disease and intravenous drug use. The routine use of invasive procedures, implantable cardiac devices and prosthetic heart valves has served to increase the number of at-risk patients. The microbiology of IE mirrors the changing risk factors, with staphylococcal infections predominating over viridans streptococci. An overview of this rare disease is given describing current understanding, investigation and changing epidemiology and microbiology of IE.


Eight years experience in crush and avulsion type finger amputation.
Between July 1986 and July 1994, we studied 125 total finger amputations due to crush and avulsion type injuries at the Izmir Hand and Microsurgery Hospital. Of the 125 fingers replanted, 88 were successful, for a general survival rate of 70.4%. In the period between 1986 and 1991, we performed end-to-end anastomoses and achieved a survival rate of 48.3% (Group I). Between 1991 and 1994, with the application of a primary interpositional vein graft for the required indications in this type of injury, the general survival rate increased to 77.1% (Group II; P < 0.05). We conclude that the significant increase in the survival rate in Group II is related to both the increased experience of the surgical team and the application of the primary interpositional vein graft.


Infectious Agents in Bovine Red Meat and Milk and Their Potential Role in Cancer and Other Chronic Diseases.
Red meat and dairy products have frequently been suggested to represent risk factors for certain cancers, chronic neurodegenerative diseases, and autoimmune and cardiovascular disorders. This review summarizes the evidence and investigates the possible involvement of infectious factors in these diseases. The isolation of small circular single-stranded DNA molecules from serum and dairy products of Eurasian Aurochs (Bos taurus)-derived cattle, obviously persisting as episomes in infected cells, provides the basis for further investigations. Gene expression of these agents in human cells has been demonstrated, and frequent infection of humans is implicated by the detection of antibodies in a high percentage of healthy individuals. Epidemiological observations suggest their relationship to the development multiple sclerosis, to heterophile antibodies, and to N-glycolylneuraminic acid (Neu5Gc) containing cell surface receptors.


National Institutes of Health Funding in Rhode Island.
We present an overview of the National Institutes of Health (NIH) funding in Rhode Island through analysis of 935 NIH grants received during the fiscal years of 2012 to 2016. NIH funded over 2,600 grants from 2012 to 2016, of which approximately 900 were new grant awards, and the remainder were annual grant renewals. The most funded type of research in Rhode Island is mental health and substance abuse, followed by infectious disease, neurology, and public health. Research funding of cardiovascular diseases, on a per capita basis, are on par with the rest of the nation, while cancer research funding is less than one half the national average. The largest NIH institutional funding source is the National Institute of General Medical Sciences (NIGMS), followed by National Institute of Mental Health (NIMH) and National Institute on Alcohol Abuse and Alcoholism (NIAAA). While research grants (R01s) remain the predominant source of NIH funding, investigators in Rhode Island have secured additional funding through program project (P) grants with the aim of bolstering research resources and collaboration throughout the state. [Full article available at http://rimed.org/rimedicaljournal-2017-07.asp].


A longitudinal study of condylar growth and mandibular rotation in untreated subjects with class II malocclusion.
Condylar growth intensity, mandibular growth rotation, and the association between growth intensity and rotation were evaluated in 32 untreated subjects with Class II malocclusions. Condylar growth was measured on serial lateral head films annually from ages 8 to 13. The analysis showed that the growth intensity of the condyles varied only slightly between girls and boys, with boys growing more. Fluctuations in growth intensity were observed in all subjects from year to year. None of the individuals maintained a consistent growth velocity over two or more consecutive years. The average condylar growth intensity was two to three mm per year throughout the period from age 8 1/2 to 12 1/2 for both sexes, with boys growing slightly faster than girls at some ages. The greatest growth velocity observed was eight mm in one year, which was seen in one boy. Three girls had as much as six mm of condylar growth in one year. Average mandibular rotation was -0.8 (+/- 0.5 SD) per year in boys and -0.6 degrees (+/- 0.6 SD) in girls, with great individual variation for both sexes. Ninety percent of the subjects showed some degree of anterior or forward rotation; only one subject demonstrated posterior rotation and three demonstrated almost no rotation. No clear relationship was found between amount of condylar growth and mandibular rotation. This study shows that condylar growth intensity varies considerably between subjects and from year to year in each subject, and suggests that clinicians need to be conservative when estimating treatment length for a patient with a Class II malocclusion.


Frequency of Dirofilaria immitis infection in blood donor dogs of the Rio de Janeiro state.
Dirofilariasis, a parasitic disease caused by the nematode Dirofilaria immitis, commonly known as heartworm, primarily inhabits the pulmonary artery and right heart of dogs and other animals. The disease is transmitted through diptera, predominantly from the Culex, Aedes, and Anopheles genera. Dirofilariasis is cosmopolitan in nature, endemic in coastal regions and tropical climates. Factors such as temperature, humidity, vector density, and the presence of definitive hosts significantly contribute to the spread of this parasitic disease. In the state of Rio de Janeiro, a prevalence of 58.6% of D. immitis infected animals has been recorded in municipalities like Niterói. Given that blood transfusions are routine clinical procedures and blood bags are not always accurately evaluated, an investigation into D. immitis infection in blood donor dogs from the metropolitan region of Rio de Janeiro was conducted. A total of 1044 blood donor dog files from a blood bank in Niterói, RJ, collected from January 2019 to December 2022, were analyzed. These samples, originating from kennels in various municipalities in the Metropolitan Region, were tested for the presence of microfilariae through direct examination using tubes and microhematocrit evaluated in optical microscopy. Additionally, the search for antigens was conducted using the enzyme-linked immunosorbent assay technique. Out of the 1044 records evaluated, 17.8% (186/1044) were positive for heartworm infection, with 2% (21/1044) samples positive for microfilariae and 14.8% (154/1044) positive for D. immitis antigens. The high prevalence rate indicates that canine D. immitis infection remains prevalent in the state of Rio de Janeiro, necessitating effective guidelines for prescribing preventive medications by veterinarians and an increase in epidemiological surveillance in the region.


Soluble and Lente human insulin mixtures in normal man.
The interaction between soluble (Actrapid HM) and Lente (Monotard HM) human insulin preparations was examined in normal subjects. Incremental plasma insulin levels were determined following the subcutaneous administration of 6 U of soluble insulin admixed with 0.14 ml soluble insulin diluting medium, or 0.14 ml Lente diluting medium and injected either immediately or 5 min after preparation. For comparison separate but simultaneous injections of soluble (6 U) and Lente insulin (14 U) were administered subcutaneously. All injection volumes were identical. The incremental insulin levels were significantly greater between 60 and 90 min (p less than 0.05-0.01) following the administration of soluble insulin admixed with its own medium compared to the soluble insulin/Lente medium admixtures injected immediately and 5 min after preparation. There was no difference, however, in the plasma insulin profiles between the two soluble insulin/Lente medium schedules. The separate simultaneous administration of soluble and Lente insulin resulted in significantly higher plasma insulin levels at 30 and 90 min (p less than 0.05) when compared to the admixture injected immediately after preparation and from 30-120 min (p less than 0.05-0.01) compared to the admixture injected after 5 min. The two soluble/Lente insulin admixtures achieved similar plasma insulin profiles. Therefore when soluble human insulin is admixed with Lente human insulin and administered by subcutaneous injection immediately after admixing there is a significant reduction in the plasma insulin levels during the first 90 min in contrast to when the two preparations are given simultaneously by separate injection. Delaying the injection of the admixture for 5 min results in significantly lower insulin levels up to 120 min. These differences are reflected in the hypoglycaemic responses observed.


Nonclosure of peritoneum: a reappraisal.
In our specialty, the practice of closing or not closing the peritoneum is still being debated. Our purpose was to review the literature on the subject, to evaluate the advantages and disadvantages of the procedure, and to provide clinical opinions. Closure of the peritoneum either parietal or visceral peritoneum is unnecessary, it is associated with a slightly longer operating time and more postoperative pain, and there are some suggestions that it might cause more adhesion formation. There are more advantages than disadvantages to not closing the peritoneum. We encourage clinicians not to close both parietal and visceral peritoneum.


Preeclampsia Status Controls Interleukin-6 and Soluble IL-6 Receptor Release from Neutrophils and Endothelial Cells: Relevance to Increased Inflammatory Responses.
Increased neutrophil-endothelial binding and inflammatory responses are significant pathophysiological events in the maternal vascular system in preeclampsia, a hypertensive disorder in human pregnancy. Interleukin 6 (IL-6) and its soluble receptors (soluble IL-6R (sIL-6R) and soluble gp130 (sgp130)) are critical inflammatory mediators. During pregnancy, maternal IL-6 and sgp130 levels were increased, but sIL-6R levels were decreased, in women with preeclampsia compared to normotensive pregnant women. However, little is known about differences in IL-6, sIL-6R, and sgp130 production by neutrophils and endothelial cells between normal pregnancy and preeclampsia. To study this, we isolated neutrophils and cultured human umbilical vein endothelial cells (HUVECs) from normal and preeclamptic pregnancies. Production of IL-6, sIL-6R, and sgp130 was measured. The role of placental factor(s)-mediated neutrophil production of IL-6, sIL-6R, and sgp130 was also determined by pretreating neutrophils with placental conditioned medium generated from placental villous cultures. We found that IL-6 and sgp130 were mainly produced by endothelial cells, while sIL-6R was mainly produced by neutrophils. Endothelial cells from preeclampsia produced significantly more IL-6 and sgp130, and neutrophils from preeclampsia produced significantly less sIL-6R than normal pregnancy cells. Interestingly, production of IL-6, sIL-6R, and sgp130 were time-dependently increased when neutrophils and endothelial cells were co-cultured. We also found that neutrophils from normal pregnancies produced more IL-6, but less sIL-6R, after being primed by preeclamptic-placental conditioned medium. These results demonstrated that neutrophils and endothelial cells have different capacities in producing IL-6, sIL-6R, and sgp130 between normal pregnancy and preeclampsia. These results also provide evidence that the placenta plays a role in inducing neutrophil activation in preeclampsia.


Complementary distal arterio-venous fistula combined with deep vein interposition to improve infrapopliteal prosthetic bypass patency.
The rapid evolution of vascular surgical techniques over the last few years coupled with the availability of an adequate venous conduit has allowed a more liberal and successful approach to salvage ischemic limbs caused by advanced atherosclerosis. This aggressive approach can be exemplified by the construction of arterial bypasses to the terminal branches of tibial vessels. However, a significant number of patients will continue to face the threat of a major amputation because of insufficient amounts of vein(s) to perform a totally autogenous bypass to one of the infrapopliteal arteries.


Depressive symptoms in women with urinary incontinence: a prospective study.
In this prospective study, the incidence of depression in women who suffer from urinary incontinence (UI) is examined. Although the literature has confounding information directly linking depression to UI, this study revealed depression was significantly higher in women with UI as opposed to their counterparts without UI.


Open-label phase I clinical trial of Ad5-EBOV in Africans in China.
To determine the safety and immunogenicity of a novel recombinant adenovirus type 5 vector based Ebola virus disease vaccine (Ad5-EBOV) in Africans in China. A phase 1, dose-escalation, open-label trial was conducted. 61 healthy Africans were sequentially enrolled, with 31 participants receiving one shot intramuscular injection and 30 participants receiving a double-shot regimen. Primary and secondary end points related to safety and immunogenicity were assessed within 28 d after vaccination. This study was registered with ClinicalTrials.gov (NCT02401373). Ad5-EBOV is well tolerated and no adverse reaction of grade 3 or above was observed. 53 (86.89%) participants reported at least one adverse reaction within 28 d of vaccination. The most common reaction was fever and the mild pain at injection site, and there were no significant difference between these 2 groups. Ebola glycoprotein-specific antibodies appeared in all 61 participants and antibodies titers peaked after 28 d of vaccination. The geometric mean titres (GMTs) were similar between these 2 groups (1919.01 vs 1684.70 P = 0.5562). The glycoprotein-specific T-cell responses rapidly peaked after 14 d of vaccination and then decreased, however, the percentage of subjects with responses were much higher in the high-dose group (60.00% vs 9.68%, P = 0.0014). Pre-existing Ad5 neutralizing antibodies could significantly dampen the specific humoral immune response and cellular response to the vaccine. The application of Ad5-EBOV demonstrated safe in Africans in China and a specific GP antibody and T-cell response could occur 14 d after the first immunization. This acceptable safety profile provides a reliable basis to proceed with trials in Africa.


Comparison of the protective effects of the pertussis acellular vaccine with the component vaccine, which have different amounts of fimbriae, against the experimental aerosol infection of mice with Bordetella pertussis.
We obtained highly purified fimbrae from Bordetella pertussis cells which gave a single band in SDS-polyacrylamide gel electrophoresis. Using the purified fimbriae, we prepared anti-fimbriae rabbit or goat IgGs by fimbriae-Sepharose affinity chromatography, and developed ELISA for its determination. The concentration of fimbriae in our former type of acellular vaccine (80 micrograms per ml) was determined to be 20 ng per ml, and that in our component vaccine was at an almost negligible level. To estimate the protective effects of the small levels of fimbriae, we evaluated the protective activities of three types of vaccines; the former type acellular vaccine (pertussis toxin (PT): filamentous hemagglutinin (FHA): fimbriae = 15:61:0.02 (microgram per ml)), the two-component vaccine (16:64:0.0001), and the three-component vaccine (16:63:0.8). The three types of vaccines showed no significant differences in protectivity against the experimental aerosol infection suggesting that these levels of fimbriae are not effective against the experimental aerosol infection of mice with Bordetella pertussis.


COVID-19 Orthopaedic Safe Care Toolset: Guidelines for the Diagnosis and Management of Patients with Fracture and COVID-19.
The SARS-CoV-2 (severe acute respiratory syndrome-coronavirus 2) was reported in Wuhan, Hubei Province, People's Republic of China, and, subsequently, in other provinces and regions across the People's Republic of China and >212 countries. COVID-19, the disease caused by this coronavirus, was declared a worldwide pandemic by the World Health Organization (WHO). The incidence of patients with fracture who are also positive for COVID-19 is on the rise. The diagnosis and management of such patients can be complicated as their clinical characteristics are heterogeneous. Furthermore, a surgical procedure can be particularly challenging given that the use of high-speed devices results in aerosol generation. In this study, we develop and propose globally applicable guidelines to fill this knowledge gap and we identify and propose the necessary protective strategies for medical personnel in an orthopaedic emergency department and in the inpatient wards. We also introduce diagnostic criteria, surgical complication management, and follow-up strategies for infected patients. These guidelines may be helpful to decrease the infection rate of orthopaedic trauma personnel and to provide diagnosis and treatment therapy for patients with fracture and COVID-19.


Metal-catalyzed double migratory cascade reactions of propargylic esters and phosphates.
Propargylic esters and phosphates are easily accessible substrates, which exhibit rich and tunable reactivities in the presence of transition metal catalysts. π-Acidic metals, mostly gold and platinum salts, activate these substrates for an initial 1,2- or 1,3-acyloxy and phosphatyloxy migration process to form reactive intermediates. These intermediates are able to undergo further cascade reactions leading to a variety of diverse structures. This tutorial review systematically introduces the double migratory reactions of propargylic esters and phosphates as a novel synthetic method, in which further cascade reaction of the reactive intermediate is accompanied by a second migration of a different group, thus offering a rapid route to a wide range of functionalized products. The serendipitous observations, as well as designed approaches involving the double migratory cascade reactions, will be discussed with emphasis placed on the mechanistic aspects and the synthetic utilities of the obtained products.


Registration of congenital anomalies in ART populations: pitfalls.
Various methodological pitfalls preclude simple validation of the ostensible safety of offspring conceived thorough assisted reproductive technologies (ART). These pitfalls include lack of an appropriate comparison group (control), failure to take into account potential confounding variables, anomaly surveillance that is either too rigorous or too lax compared to the way in which anomalies are sought in general population surveys, and inconsistent criteria with respect to classifying anomalies (inclusion or exclusion of minor anomalies, internal anomalies evident only on ultrasound, and anomalies present in terminated pregnancies). To minimise these pitfalls, we recommend prospective surveillance for major anomalies, best defined as defects resulting in death, causing major handicap or necessitating surgery. Surveillance should ideally begin as soon as pregnancy is diagnosed. Data should be gathered allowing potential confounding variables to be taken into account, and systematic neonatal surveillance for anomalies should be performed during a designated time interval.


Deep-Sea Bacterium Shewanella piezotolerans WP3 Has Two Dimethyl Sulfoxide Reductases in Distinct Subcellular Locations.
Dimethyl sulfoxide (DMSO) acts as a substantial sink for dimethyl sulfide (DMS) in deep waters and is therefore considered a potential electron acceptor supporting abyssal ecosystems. Shewanella piezotolerans WP3 was isolated from west Pacific deep-sea sediments, and two functional DMSO respiratory subsystems are essential for maximum growth of WP3 under in situ conditions (4°C/20 MPa). However, the relationship between these two subsystems and the electron transport pathway underlying DMSO reduction by WP3 remain unknown. In this study, both DMSO reductases (type I and type VI) in WP3 were found to be functionally independent despite their close evolutionary relationship. Moreover, immunogold labeling of DMSO reductase subunits revealed that the type I DMSO reductase was localized on the outer leaflet of the outer membrane, whereas the type VI DMSO reductase was located within the periplasmic space. CymA, a cytoplasmic membrane-bound tetraheme c-type cytochrome, served as a preferential electron transport protein for the type I and type VI DMSO reductases, in which type VI accepted electrons from CymA in a DmsE- and DmsF-independent manner. Based on these results, we proposed a core electron transport model of DMSO reduction in the deep-sea bacterium S. piezotolerans WP3. These results collectively suggest that the possession of two sets of DMSO reductases with distinct subcellular localizations may be an adaptive strategy for WP3 to achieve maximum DMSO utilization in deep-sea environments.IMPORTANCE As the dominant methylated sulfur compound in deep oceanic water, dimethyl sulfoxide (DMSO) has been suggested to play an important role in the marine biogeochemical cycle of the volatile anti-greenhouse gas dimethyl sulfide (DMS). Two sets of DMSO respiratory systems in the deep-sea bacterium Shewanella piezotolerans WP3 have previously been identified to mediate DMSO reduction under in situ conditions (4°C/20 MPa). Here, we report that the two DMSO reductases (type I and type VI) in WP3 have distinct subcellular localizations, in which type I DMSO reductase is localized to the exterior surface of the outer membrane and type VI DMSO reductase resides in the periplasmic space. A core electron transport model of DMSO reduction in WP3 was constructed based on genetic and physiological data. These results will contribute to a comprehensive understanding of the adaptation mechanisms of anaerobic respiratory systems in benthic microorganisms.


Identification and characteristics of the testes-specific gene, Ccdc38, in mice.
Distinguishing the testes-specific genes in different species may disclose key genes associated with testes-specific functions and provide sufficient information for the study and treatment of male infertility. A testes‑specific gene, coiled-coil domain containing 38 (Ccdc38), was identified by screening UniGene libraries. Systematic bioinformatics analysis demonstrated that the CCDC38 protein was conserved in various mammalian species. It was determined that CCDC38 was exclusively expressed in testes and its expression increased from 2‑8 weeks of age. Additional immunohistochemical analysis indicated that CCDC38 was mainly expressed in spermatogonia and spermatocytes. It is of note that, immunofluorescence and co-immunoprecipitation assays demonstrated that CCDC38 interacted with ubiquitinated histone H2A in mouse testes. Therefore, these results suggest that Ccdc38 is a testes-specific gene, which may be important for mouse spermatogenesis.


Effects of monensin and colchicine on myelin galactolipids.
Monensin and colchicine have been used in a variety of systems to disrupt functioning of the Golgi apparatus and transport of Golgi-derived vesicles to the plasma membrane. In this study the effects of monensin and colchicine on the synthesis of cerebroside and sulfatide and their appearance in myelin were examined to determine whether these myelin components are processed through the Golgi apparatus. Brain slices from rats 17 days old were incubated with [3H]galactose and [35S]-sulfate to label cerebroside and sulfatide. Myelin was isolated on sucrose density gradients. Fractions highly enriched in cerebroside and sulfatide were prepared from homogenates and myelin fractions by lipid extraction, alkaline methanolysis, and in some cases TLC. Monensin at 0.1 microM had no significant effect on synthesis of these galactolipids as measured by incorporation of [3H]-galactose into cerebroside or [35S]sulfate into sulfatide in homogenates. However, appearance of [35S]sulfatide in the myelin fraction was reduced to 49% of control, while appearance of [3H]cerebroside was not significantly reduced. Colchicine from 1 mM to 0.1 microM had effects similar to monensin, that is, appearance of [35S]sulfatide in myelin was depressed, but again [3H]cerebroside was not affected. Incorporation of [35S]sulfate into sulfatide in homogenate was 93% of control, while appearance of [35S]sulfatide in the myelin fraction was depressed to 58% of control. The inhibition of appearance of sulfatide in myelin by colchicine and monensin is consistent with the view that sulfation of cerebroside occurs in the Golgi and that sulfatide is transported via Golgi-derived vesicles to the forming myelin membrane.(ABSTRACT TRUNCATED AT 250 WORDS)


Development in rice genome research based on accurate genome sequence.
Rice is one of the most important crops in the world. Although genetic improvement is a key technology for the acceleration of rice breeding, a lack of genome information had restricted efforts in molecular-based breeding until the completion of the high-quality rice genome sequence, which opened new opportunities for research in various areas of genomics. The syntenic relationship of the rice genome to other cereal genomes makes the rice genome invaluable for understanding how cereal genomes function. Producing an accurate genome sequence is not an easy task, and it is becoming more important as sequence deviations among, and even within, species highlight functional or evolutionary implications for comparative genomics.


Biologically-equivalent dose and long-term survival time in radiation treatments.
Within the linear-quadratic model the biologically-effective dose (BED)-taken to represent treatments with an equal tumor control probability (TCP)-is commonly (and plausibly) calculated according to BED(D) = -log[S(D)]/alpha. We ask whether in the presence of cellular proliferation this claim is justified and examine, as a related question, the extent to which BED approximates an isoeffective dose (IED) defined, more sensibly, in terms of an equal long-term survival probability, rather than TCP. We derive, under the assumption that cellular birth and death rates are time homogeneous, exact equations for the isoeffective dose, IED. As well, we give a rigorous definition of effective long-term survival time, T(eff). By using several sets of radiobiological parameters, we illustrate potential differences between BED and IED on the one hand and, on the other, between T(eff) calculated as suggested here or by an earlier recipe. In summary: (a) the equations currently in use for calculating the effective treatment time may underestimate the isoeffective dose and should be avoided. The same is the case for the tumor control probability (TCP), only more so; (b) for permanent implants BED may be a poor substitute for IED; (c) for a fractionated treatment schedule, interpreting the observed probability of cure in terms of a TCP formalism that refers to the end of the treatment (rather than T(eff)) may result in a miscalculation (underestimation) of the initial number of clonogens.


Health economic analysis of human papillomavirus vaccines in women of Chile: perspective of the health care payer using a Markov model.
In Chile, significant reductions in cervical cancer incidence and mortality have been observed due to implementation of a well-organized screening program. However, it has been suggested that the inclusion of human papillomavirus (HPV) vaccination for young adolescent women may be the best prospect to further reduce the burden of cervical cancer. This cost-effectiveness study comparing two available HPV vaccines in Chile was performed to support decision making on the implementation of universal HPV vaccination. The present analysis used an existing static Markov model to assess the effect of screening and vaccination. This analysis includes the epidemiology of low-risk HPV types allowing for the comparison between the two vaccines (HPV-16/18 AS04-adjuvanted vaccine and the HPV-6/11/16/18 vaccine), latest cross-protection data on HPV vaccines, treatment costs for cervical cancer, vaccine costs and 6% discounting per the health economic guideline for Chile. Projected incremental cost-utility ratio (ICUR) and incremental cost-effectiveness ratio (ICERs) for the HPV-16/18 AS04-adjuvanted vaccine was 116 United States (US) dollars per quality-adjusted life years (QALY) gained or 147 US dollars per life-years (LY) saved, while the projected ICUR/ICER for the HPV-6/11/16/18 vaccine was 541 US dollars per QALY gained or 726 US dollars per LY saved. Introduction of any HPV vaccine to the present cervical cancer prevention program of Chile is estimated to be highly cost-effective (below 1X gross domestic product [GDP] per capita, 14278 US dollars). In Chile, the addition of HPV-16/18 AS04-adjuvanted vaccine to the existing screening program dominated the addition of HPV-6/11/16/18 vaccine. In the probabilistic sensitivity analysis results show that the HPV-16/18 AS04-adjuvanted vaccine is expected to be dominant and cost-saving in 69.3% and 77.6% of the replicates respectively. The findings indicate that the addition of any HPV vaccine to the current cervical screening program of Chile will be advantageous. However, this cost-effectiveness model shows that the HPV-16/18 AS04-adjuvanted vaccine dominated the HPV-6/11/16/18 vaccine. Beyond the context of Chile, the data from this modelling exercise may support healthcare policy and decision-making pertaining to introduction of HPV vaccination in similar resource settings in the region.


Quantitative and qualitative sex differences in spatial navigation.
We examined sex differences in spatial navigation performance using an ecologically relevant experimental paradigm in which virtual maze-like museums are projected in front of a treadmill. Thirty-two 20-30-year-old adults (16 women/16 men) performed a way-finding task in city-block (straight corridors) or variable (irregular corridors) topographies while walking on the treadmill. Sex differences in spatial navigation performance were reduced in variable topographies, suggesting less reliance on spatial relational learning among women. Also, spatial geometric knowledge of the mazes continued to be higher in men after all participants had attained perfect place-finding performance. Results indicate that sex differences in spatial navigation performance are modulated by interactions between environmental demands and sex differences in spatial processing.


Expression of an unusual T cell receptor (TCR)-V beta repertoire by Ly-6C+ subpopulations of CD4+ and/or CD8+ thymocytes. Evidence for a developmental relationship between Ly-6C+ thymocytes and CD4-CD8-TCR-alpha beta+ thymocytes.
A novel thymocyte subpopulation expressing an unusual TCR repertoire was identified by high surface expression of the Ly-6C Ag. Ly-6C+ thymocytes were distributed among all four CD4/CD8 thymocyte subsets, and represented a readily identifiable subpopulation within each one. Ly-6C+ thymocytes express TCR-alpha beta, arise late in ontogeny, and appear in the CD4/CD8 developmental pathway after birth in a sequence that resembles that followed by conventional Ly-6C- cells during fetal ontogeny. Most interestingly, adult Ly-6C+ thymocytes express an unusual TCR-V beta repertoire that is identical to that expressed by CD4-CD8-TCR-alpha beta+ thymocytes in its overexpression of TCR-V beta 8 and in its expression of some potentially autoreactive TCR-V beta specificities. This unusual TCR-V beta repertoire was even expressed by Ly-6C+ thymocytes contained within the CD4+ CD8- 'single positive' thymocyte subset. Thus, expression of this unusual TCR-V beta repertoire is not limited to CD4-CD8-thymocytes, and is unlikely to be a consequence of their double negative phenotype. Rather, we think that Ly-6C+TCR-alpha beta+ thymocytes and CD4-CD8-TCR-alpha beta+ are developmentally interrelated, a conclusion supported by several lines of evidence including the selective failure of both Ly-6C+ and CD4-CD8-TCR-alpha beta+ thymocyte subsets to appear in TCR-beta transgenic mice. In contrast, peripheral Ly-6C+ T cells are developmentally distinct from Ly-6C+ thymocytes in that peripheral Ly-6C+ T cells expressed a conventional TCR-V beta repertoire and developed normally in TCR-beta transgenic mice in which Ly-6C+ thymocytes failed to arise. We conclude that: 1) expression of a skewed TCR-V beta repertoire is a characteristic of Ly-6C+TCR-alpha beta+ thymocytes as well as CD4-CD8-TCR-alpha beta+ thymocytes, and is not unique to thymocytes expressing neither CD4 nor CD8 accessory molecules; and 2) Ly-6C+ thymocytes are developmentally linked to CD4-CD8-TCR-alpha beta+ thymocytes, but not to Ly-6C+ peripheral T cells. We suggest that Ly-6C+TCR-alpha beta+ thymocytes are not the developmental precursors of Ly-6C+ peripheral T cells, but rather may be the developmental precursors of CD4-CD8-TCR-alpha beta+ thymocytes.


Towards a Generic Connection of EHR and DSS.
Decision Support Systems (DSS) are typically integrated in Electronic Health Record systems (EHR). By removing this integration full reuse of a DSS system is possible. The connection between the EHR and the DSS system should be standards-based and generic. We intend to demonstrate the viability of this setup by implementing it using PropeRWeb as EHR system, Gaston as DSS system, HL7v3 messages and the SOAP protocol.


[Role of kidney in calcium homeostasis and premature aging].
The kidney plays a critical role in the calcium (Ca) homeostasis. With advancing age, the kidney function, including glomerular filtration rate and tubular electrolyte reabsorption, is deteriorating without any kidney disease. Disturbed Ca metabolism may be associated with premature aging process, in which various cellular function is impaired by intracellular Ca burden due to increased Ca efflux from bone. Fibroblast growth factor (FGF) 23, a novel phosphate (P) -regulating factor, is involved in disorders of mineral bone metabolism. Recent studies suggest that FGF23 plays a certain role in premature aging process through altered P and vitamin D metabolism.


A Decision-Based Modified Total Variation Diffusion Method for Impulse Noise Removal.
Impulsive noise removal usually employs median filtering, switching median filtering, the total variation L1 method, and variants. These approaches however often introduce excessive smoothing and can result in extensive visual feature blurring and thus are suitable only for images with low density noise. A new method to remove noise is proposed in this paper to overcome this limitation, which divides pixels into different categories based on different noise characteristics. If an image is corrupted by salt-and-pepper noise, the pixels are divided into corrupted and noise-free; if the image is corrupted by random valued impulses, the pixels are divided into corrupted, noise-free, and possibly corrupted. Pixels falling into different categories are processed differently. If a pixel is corrupted, modified total variation diffusion is applied; if the pixel is possibly corrupted, weighted total variation diffusion is applied; otherwise, the pixel is left unchanged. Experimental results show that the proposed method is robust to different noise strengths and suitable for different images, with strong noise removal capability as shown by PSNR/SSIM results as well as the visual quality of restored images.


Multiorgan segmentation using distance-aware adversarial networks.
Segmentation of organs at risk (OAR) in computed tomography (CT) is of vital importance in radiotherapy treatment. This task is time consuming and for some organs, it is very challenging due to low-intensity contrast in CT. We propose a framework to perform the automatic segmentation of multiple OAR: esophagus, heart, trachea, and aorta. Different from previous works using deep learning techniques, we make use of global localization information, based on an original distance map that yields not only the localization of each organ, but also the spatial relationship between them. Instead of segmenting directly the organs, we first generate the localization map by minimizing a reconstruction error within an adversarial framework. This map that includes localization information of all organs is then used to guide the segmentation task in a fully convolutional setting. Experimental results show encouraging performance on CT scans of 60 patients totaling 11,084 slices in comparison with other state-of-the-art methods.


A Review of Atrial Fibrillation Detection Methods as a Service.
Atrial Fibrillation (AF) is a common heart arrhythmia that often goes undetected, and even if it is detected, managing the condition may be challenging. In this paper, we review how the RR interval and Electrocardiogram (ECG) signals, incorporated into a monitoring system, can be useful to track AF events. Were such an automated system to be implemented, it could be used to help manage AF and thereby reduce patient morbidity and mortality. The main impetus behind the idea of developing a service is that a greater data volume analyzed can lead to better patient outcomes. Based on the literature review, which we present herein, we introduce the methods that can be used to detect AF efficiently and automatically via the RR interval and ECG signals. A cardiovascular disease monitoring service that incorporates one or multiple of these detection methods could extend event observation to all times, and could therefore become useful to establish any AF occurrence. The development of an automated and efficient method that monitors AF in real time would likely become a key component for meeting public health goals regarding the reduction of fatalities caused by the disease. Yet, at present, significant technological and regulatory obstacles remain, which prevent the development of any proposed system. Establishment of the scientific foundation for monitoring is important to provide effective service to patients and healthcare professionals.


Pregnancy With SLE and Fetal Congenital Heart Block: A Case Report.
Autoimmune AV block is usually seen in association with autoimmune antibodies in mother that cross the placenta and damage the AV node of fetus. A 24-year-old primigravida, diagnosed to have SLE, at 25 weeks period of gestation found to have fetal bradycardia. Her ANA was moderately positive, SS-A (Ro) antibodies and SS-B (La) antibodies were positive. Fetal ECHO showed no structural defect but heart rate was 55 - 60 beats per minute. She was put on dexamethasone (4 mg/day). She was lost on follow up and presented at term in emergency with labor pains and fetal bradycardia, underwent a lower segment caesarean section. Baby underwent a temporary cardiac pacing within 10 hours of birth followed by permanent pacing on day 3 of birth. Baby is doing well on follow up. Neonates with isolated congenital heart block who are monitored antenatally and delivered in a planned fashion at an institution capable of early pacing can have favorable outcomes.


The Griqua of Campbell, Cape Provice, South Africa.
The Griqua of Campbell in the Cape Province of South Africa are reputed to be descended from an amalgam of Khoi ("Hottentots") with various Caucasoid, Negroid and East Asian elements at the Cape of Good Hope. A stormy history of migration and deprivation has left this small residuum on the edge of the Kalahari Desert. It was hoped that a sero-genetic study of these people, who proudly preserve their identity, might provide further insights into the genetic consitution of their Khoi forebears. Despite the retention of the language of their own, an social mores distinct from of other Mixed populations, they have been shown, however, to have received an appreciable inflow of non-Khoisan genes. This has probably resulted from social factors such as the prestige attaching to Griqua membership and the willingness of the Griqua to assimilate outsiders, combined with the tendency of Campbell Griqua to migrate elsewhere. There are no significant differences between "Griqua" members of the population and those who are not recognized as "Griqua."


A prospective study of adiposity and all-cause mortality: the Malmö Diet and Cancer Study.
This study aims to examine the association between various measures of adiposity and all-cause mortality in Swedish middle-aged and older men and women and, additionally, to describe the influences of age and sex on these associations. A prospective analysis was performed in a cohort of 10,902 men and 16,814 women ages 45 to 73 years who participated in the Malmö Diet and Cancer Study in Sweden. Baseline examinations took place between 1991 and 1996, and 982 deaths were documented during an average follow-up of 5.7 years. All-cause mortality was related to the following variables measured at baseline: body mass index (BMI), percentage of body fat, lean body mass (LBM), and waist-to-hip ratio (WHR), with adjustment for age and selected covariates. Body composition data were derived from bioelectrical impedance analysis. The association between percentage of body fat and mortality was modified by age, particularly in women. For instance, fatness was associated with excess mortality in the younger women but with reduced mortality in the older women. Weaker associations were seen for BMI than for percentage of body fat in both sexes. Placement in the top quintiles of waist-to-hip ratio, independent of overall body fat, was a stronger predictor of mortality in women than in men. The observed associations could not be explained by bias from early death or antecedent disease. The findings reveal sex and age differences for the effects of adiposity and WHR on mortality and indicate the importance of considering direct measures of adiposity, as opposed to BMI, when describing obesity-related mortality risks.


Deep brain stimulation complicated by bilateral large cystic cavitation around the leads in a patient with Parkinson's disease.
Deep brain stimulation (DBS) is an approved and effective therapy for patients suffering from advanced Parkinson's disease (PD). Several clinical trials have indicated significant motor function improvement in patients undergoing subthalamic nucleus stimulation. This therapy is, rarely, associated with complications, mostly related to infections, seizures or stimulation-induced side effects. We report a case of a 71-year-old man with a 10-year history of PD who underwent bilateral placement of subthalamic nucleus DBS. As a complication, the patient showed subjective postoperative cognitive decline, and subsequent MRI showed peri-lead oedema, which progressed to large cystic cavitation around the leads without indication of infection. The patient received steroid therapy and the cavitations regressed without surgical intervention.


[Emergence of methicillin-resistant Staphylococcus aureus SCCmec type IV/V epidemic clones in a large teaching hospital in China].
To investigate the staphylococcal cassette chromosome mec (SCCmec) genotype and molecular epidemiological characteristics of healthcare-associated methicillin-resistant Staphylococcus aureus (HA-MRSA) in a large teaching hospital in China. From January 2012 to December 2012, a total of 71 nonduplicate HA-MRSA were collected in a teaching hospital in Changsha, China. SCCmec types were determined by multiplex PCR, and Panton-Valentine leukocidin (PVL) gene was detected by PCR. The homology among the tested isolates was determined using pulsed-field gel electrophoresis (PFGE). Of the 71 HA-MRSA isolates, 49 (69.0%) carried SCCmec III, 10 (14.1%) carried SCCmec IV, 3 (4.2%) carried SCCmec V and 3 (4.2%) carried SCCmec II; the remaining 6 isolates were not typeable by PCR. Compared with patients having SCCmec I/II/III MRSA infections, those with SCCmec IV/V MRSA infections had a significantly younger age and a similar duration of hospital stay before the first MRSA-positive culture and total hospital stay. PVL genes were strongly associated with SCCmec type IV/V MRSA infections. HA-SCCmec IV/V MRSA strains showed a greater susceptibility to rifampicin, gentamicin, levofloxacin, ciprofloxacin, and tetracycline than HA-SCCmec I/II/III MRSA strains. The 13 HA-SCCmec IV/V MRSA isolates formed one large group at the 55% similarity level. Three PFGE clusters with a similarity index of 85% or more were identified, and unique PFGE profiles were observed in 4 isolates. This is the first report of HA-MRSA isolates carrying SCCmec V in Chinese hospitals. SCCmec types IV and V MRSA clones have emerged in Chinese hospitals, which urges more rigorous surveillance of their spread in healthcare facilities in China.


Sequence Determinants of Substrate Ambiguity in a HAD Phosphosugar Phosphatase of Arabidopsis Thaliana.
The Arabidopsis thaliana broad-range sugar phosphate phosphatase AtSgpp (NP_565895.1, locus AT2G38740) and the specific DL-glycerol-3-phosphatase AtGpp (NP_568858.1, locus AT5G57440) are members of the wide family of magnesium-dependent acid phosphatases subfamily I with the C1-type cap domain haloacid dehalogenase-like hydrolase proteins (HAD). Although both AtSgpp and AtGpp have a superimporsable α/β Rossmann core active site, they differ with respect to the loop-5 of the cap domain, accounting for the differences in substrate specificity. Recent functional studies have demonstrated the essential but not sufficient role of the signature sequence within the motif-5 in substrate discrimination. To better understand the mechanism underlying the control of specificity, we explored additional sequence determinants underpinning the divergent evolutionary selection exerted on the substrate affinity of both enzymes. The most evident difference was found in the loop preceding the loop-5 of the cap domain, which is extended in three additional residues in AtSgpp. To determine if the shortening of this loop would constrain the substrate ambiguity of AtSgpp, we deleted these three aminoacids. The kinetic analyses of the resulting mutant protein AtSgpp3Δ (ΔF53, ΔN54, ΔN55) indicate that promiscuity is compromised. AtSgpp3Δ displays highest level of discrimination for D-ribose-5-phosphate compared to the rest of phosphate ester metabolites tested, which may suggest a proper orientation of D-ribose-5-phosphate in the AtSgpp3Δ active site.


Epithelial-to-mesenchymal transition involves triacylglycerol accumulation in DU145 prostate cancer cells.
Epithelial to mesenchymal transition (EMT) is a biological process that plays a crucial role in cancer metastasis. Although studies regarding the EMT mechanisms are usual in terms of gene expression and protein functions, little is known about the involvement of lipids in EMT. In this work, an untargeted lipidomic analysis was performed to reveal which lipids are involved in the EMT process. DU145 prostate cancer cells were treated with TNFα, a well-known EMT inducer. After 6 hours of treatment, a decrease of cell membrane E-cadherin as well as a reduction in its gene expression were observed. Also, the mesenchymal markers Vimentin and Snail were up-regulated, suggesting that EMT started below 6 hours of treatment. Lipid extracts of untreated and TNFα-treated cells at short times were analyzed using ultra-performance liquid chromatography coupled to high-resolution mass spectrometry (UPLC-MS). Multivariate data analysis methods were applied to decipher which lipids presented significant changes after EMT induction. Among the results obtained, a significant increase of twelve unsaturated triacylglycerides (TAGs) was observed. This increase of TAGs was also observed for cells treated with TGFβ (another EMT inducer), suggesting that this feature is a common mechanism in the EMT process. In conclusion, this work reported for the first time a TAG accumulation through EMT induction. These TAG lipids could play a key role in providing cells with the energy, cell membrane components and signaling lipids necessary to guarantee the enhanced cell migration and proliferation of metastatic cells.


Impact of an augmented intervention on self-regulatory, dietary and physical activity outcomes in a diabetes prevention trial among adults with prediabetes.
Self-regulation can facilitate modifications in lifestyle to promote behavioral change. However, little is known about whether adaptive interventions promote improvement in self-regulatory, dietary, and physical activity outcomes among slow treatment responders. A stratified design with an adaptive intervention for slow responders was implemented and evaluated. Adults ≥ 21 years old with prediabetes were stratified to the standard Group Lifestyle Balance intervention (GLB; n = 79) or the adaptive GLB Plus intervention (GLB + ; n = 105) based on first-month treatment response. Intake of total fat was the only study measure that significantly differed between groups at baseline (P = 0.0071). GLB reported greater improvement in self-efficacy for lifestyle behaviors, goal satisfaction with weight loss, and very active minutes of activity than GLB + (all P < 0.01) at 4-months. Both groups reported significant improvement in self-regulatory outcomes and reduction in energy and fat intake (all P < 0.01). An adaptive intervention can improve self-regulation and dietary intake when tailored to early slow treatment responders.


Situation analysis of child labour in Karachi, Pakistan: a qualitative study.
In Karachi, large employment opportunities, burgeoning population and the availability of cheap labour might be the contributing factors for the increasing prevalence of child labour. A literature review was conducted in 2007 that included published and unpublished literature since 2000. Various organizations working in the field were also covered, while the perception of the child labourers was covered through three focus group discussions. Common health issues among the child labourers in Karachi included respiratory illnesses, fever and generalised pains, as well as drug and sexual abuse. Organisations working for child labour could be broadly categorised into those working for legal advice and advocacy; those generating statistics; and those that are providing interventions. Discussion with children showed that irrespective of the immediate cause, the underlying determinant for child labour was poverty. The best practices identified included evening schools and drop-in centers for working children with provision for skill-based education and basic health facilities. There is need to have more such centres.


A case of atypical dengue haemorrhagic fever.
A case of atypical dengue haemorrhagic fever is being described in a 30 years old male along with a short discussion on the subject.


TMS and neocortical neurons: an integrative review on the micro-macro connection in neuroplasticity.
Tian D, Izumi S. TMS and neocortical neurons: an integrative review on the micro-macro connection in neuroplasticity. Jpn J Compr Rehabil Sci 2023; 14: 1-9. Neuroplasticity plays a pivotal role in neuroscience and neurorehabilitation as it bridges the organization and reorganization properties of the brain. Among the numerous neuroplastic protocols, transcranial magnetic stimulation (TMS) is a well-established non-invasive protocol to induce plastic changes in the brain. Here, we review the findings of four plasticity-inducing TMS protocols in the human motor cortex with relatively evident mechanisms: conventional repetitive TMS (rTMS), theta-burst stimulation (TBS), quadripulse stimulation (QPS) and paired associative stimulation (PAS). Based on the reviewed evidence and a preliminary TMS neurocytological model proposed in our previous report, we further integrate the neurophysiological evidence and plasticity rules of these protocols to present an updated micro-macro connection model between neocortical neurons and the neurophysiological evidence in TMS. This prototypical model will guide further efforts to understand the neural circuit of the motor cortex, the mechanisms of TMS, and the advance of neuroplasticity technologies and their outcomes.


[Type IV Ehlers-Danlos syndrome associated with mitral valve prolapse: a case report].
A 52-year-old male patient with the Ehlers-Danlos syndrome (familial hypermobility of the joints, hyperextensibility of the skin and atrophic cutaneous scars) was evaluated because of a mitral regurgitant murmur. Echocardiography demonstrated marked mitral valve prolapse of the both leaflets and vegetation-like thickening of the anterior leaflet of the mitral valve. Two-dimensional color flow mapping showed severe mitral regurgitation. This patient had also acrogeria-like facial appearance and very thin skin (subcutaneous veins were readily visible). He developed repeated rupture of radial, ulnar and middle cerebral arteries and expired. A histological section of the ruptured ulnar artery demonstrated no infectious process. In view of the joint hyperextensibility, very thin skin, characteristic facial appearance and the spontaneous occurrence of three successive ruptures of peripheral arteries, a diagnosis of Ehlers-Danlos syndrome (type IV) was made. Although mitral valve prolapse is reportedly common in type I, II and III Ehlers-Danlos syndrome, this report emphasizes that it may also occur in patients with type IV Ehlers-Danlos syndrome and the relevant literatures are reviewed.


Altered cell cycle phase distributions in cultured human carcinoma cells partially depleted of polyamines by treatment with difluoromethylornithine.
We have studied the effects of partial polyamine depletion, induced by treatment with alpha-difluoromethylornithine (DFMO) on cell cycle phase distributions in five cultured human carcinoma cell lines. We used flow cytometry of cells stained with chromomycin-A3 and computer analysis to measure phase distributions of treated and control cultures. All five lines respond to 1-5 mM DFMO treatment with a total absence of measurable putrescine, a loss of greater than 90% of spermidine, and a 30-40% decline in spermine by 48 h after DFMO addition. The proliferation of all five lines is inhibited as well. Nonetheless, only four of the cell lines (HuTu-80, HT-29, MCF-7, and A-427) show a marked increase in the G1-phase fraction and decrease in the S-phase fraction as a consequence of DFMO treatment. Small, but significant, decreases in the G2-M populations of these cell lines also occurred after DFMO treatment. Exogenous putrescine (5-50 microM) reversed both the polyamine depletion and the perturbed phase distributions of DFMO-treated cultures but was without effect on phase distributions of cultures not treated with DFMO. The fifth cell line (ME-180) showed no effect of polyamine depletion on cell cycle phase distributions in DFMO-treated cultures and also no effect of exogenous putrescine on phase fractions of either control or DFMO-treated cells. These observations indicate that some human tumor cell lines are dependent upon adequate intracellular polyamine content for maintenance of cell cycle traverse. They also imply that human tumor cell lines are heterogeneous with regard to their cell cycle response to DFMO-induced polyamine deficiency.


Mathematical Model of Naive T Cell Division and Survival IL-7 Thresholds.
We develop a mathematical model of the peripheral naive T cell population to study the change in human naive T cell numbers from birth to adulthood, incorporating thymic output and the availability of interleukin-7 (IL-7). The model is formulated as three ordinary differential equations: two describe T cell numbers, in a resting state and progressing through the cell cycle. The third is introduced to describe changes in IL-7 availability. Thymic output is a decreasing function of time, representative of the thymic atrophy observed in aging humans. Each T cell is assumed to possess two interleukin-7 receptor (IL-7R) signaling thresholds: a survival threshold and a second, higher, proliferation threshold. If the IL-7R signaling strength is below its survival threshold, a cell may undergo apoptosis. When the signaling strength is above the survival threshold, but below the proliferation threshold, the cell survives but does not divide. Signaling strength above the proliferation threshold enables entry into cell cycle. Assuming that individual cell thresholds are log-normally distributed, we derive population-average rates for apoptosis and entry into cell cycle. We have analyzed the adiabatic change in homeostasis as thymic output decreases. With a parameter set representative of a healthy individual, the model predicts a unique equilibrium number of T cells. In a parameter range representative of persistent viral or bacterial infection, where naive T cell cycle progression is impaired, a decrease in thymic output may result in the collapse of the naive T cell repertoire.


Senescent Stromal Cells Promote Cancer Resistance through SIRT1 Loss-Potentiated Overproduction of Small Extracellular Vesicles.
Cellular senescence is a potent tumor-suppressive program that prevents neoplastic events. Paradoxically, senescent cells develop an inflammatory secretome, termed the senescence-associated secretory phenotype, which is implicated in age-related pathologies including cancer. Here, we report that senescent cells actively synthesize and release small extracellular vesicles (sEV) with a distinctive size distribution. Mechanistically, SIRT1 loss supported accelerated sEV production despite enhanced proteome-wide ubiquitination, a process correlated with ATP6V1A downregulation and defective lysosomal acidification. Once released, senescent stromal sEVs significantly altered the expression profile of recipient cancer cells and enhanced their aggressiveness, specifically drug resistance mediated by expression of ATP-binding cassette subfamily B member 4 (ABCB4). Targeting SIRT1 with agonist SRT2104 prevented development of cancer resistance by restraining sEV production by senescent stromal cells. In clinical oncology, sEVs in peripheral blood of posttreatment cancer patients were readily detectable by routine biotechniques, presenting an exploitable biomarker to monitor therapeutic efficacy and predict long-term outcome. Together, this study identifies a distinct mechanism supporting pathologic activities of senescent cells and provides a potent avenue to circumvent advanced human malignancies by cotargeting cancer cells and their surrounding microenvironment, which contributes to drug resistance via secretion of sEVs from senescent stromal cells. SIGNIFICANCE: Senescent stromal cells produce a large number of sEVs to promote cancer resistance in therapeutic settings, a process driven by SIRT1 decline in stromal cells and ABCB4 augmentation in cancer cells.See related commentary by Wiley, p. 3193 GRAPHICAL ABSTRACT: http://cancerres.aacrjournals.org/content/canres/80/16/3383/F1.large.jpg.


Immunohistochemical studies of peptidergic neurons in the dorsal horn of the spinal cord.
The indirect immunofluorescence technique was used to localize substance P, somatostatin, methionine--enkephalin, neurotensin, and oxytocin in the dorsal horn of the rat spinal cord. The unique distribution of each peptide is described and the relative amount of each peptide in laminae I--III of the dorsal horn and the dorsal part of the lateral funniculus qualitatively assessed. Colchicine treatment and dorsal rhizotomy were used to determine, in part, the origin of immunoreactive fibers and terminals observed in the dorsal horn.


Selenium Analysis and Speciation in Dietary Supplements Based on Next-Generation Selenium Ingredients.
Selenium is essential for humans and the deficit of Se requires supplementation. In addition to traditional forms such as Se salts, amino acids, or selenium-enriched yeast supplements, next-generation selenium supplements, with lower risk for excess supplementation, are emerging. These are based on selenium forms with lower toxicity, higher bioavailability, and controlled release, such as zerovalent selenium nanoparticles (SeNPs) and selenized polysaccharides (SPs). This article aims to focus on the existing analytical systems for the next-generation Se dietary supplement, providing, at the same time, an overview of the analytical methods available for the traditional forms. The next-generation dietary supplements are evaluated in comparison with the conventional/traditional ones, as well as the analysis and speciation methods that are suitable to reveal which Se forms and species are present in a dietary supplement. Knowledge gaps and further research potential in this field are highlighted. The review indicates that the methods of analysis of next-generation selenium supplements should include a step related to chemical species separation. Such a step would allow a proper characterization of the selenium forms/species, including molecular mass/dimension, and substantiates the marketing claims related to the main advantages of these new selenium ingredients.


Biosynthesis of chlorosome proteins is not inhibited in acetylene-treated cultures of Chlorobium vibrioforme.
The composition, abundance and apparent molecular masses of chlorosome polypeptides from Chlorobium tepidum and Chlorobium vibrioforme 8327 were compared. The most abundant, low-molecular-mass chlorosome polypeptides of both strains had similar electrophoretic mobilities and abundances, but several of the larger proteins were different in both apparent mass and abundance. Polyclonal antisera raised against recombinant chlorosome proteins of Cb. tepidum recognized the homologous proteins in Cb. vibrioforme, and a one-to-one correspondence between the chlorosome proteins of the two species was confirmed. As previously shown [Ormerod et al. (1990) J Bacteriol 172: 1352-1360], acetylene strongly suppressed the synthesis of bacteriochlorophyll c in Cb. vibrioforme strain 8327. No correlation was found between the bacteriochlorophyll c content of cells and the cellular content of chlorosome proteins. Nine of ten chlorosome proteins were detected in acetylene-treated cultures, and the chlorosome proteins were generally present in similar amounts in control and acetylene-treated cells. These results suggest that the synthesis of chlorosome proteins and the assembly of the chlorosome envelope is constitutive. It remains possible that the synthesis of bacteriochlorophyll c and its insertion into chlorosomes might be regulated by environmental parameters such as light intensity.


Evolution of a Natural Products and Nutraceuticals Course in the Pharmacy Curriculum.
Objective. To develop, implement, and modify a required, second-year pharmacy course that provides an understanding of the scientific, therapeutic, and clinical principles, as well as the evidence-based medicine underlying the use of natural products. Design. A 28-hour, multi-faculty course was developed and offered in 2008. The course was modified over the years to enhance students' practice skills in the use of natural products. A course evaluation and survey were administered to assess the students' opinions. Assessment. Students performed well in the course and provided favorable evaluations, especially for the latest offering. Students reported significantly improved skills in providing advice to patients regarding the use of natural products. Conclusion. The course increased the students' knowledge and application of information and counseling skills regarding natural products.


Cigarette smoking and health-promoting behaviours among tuberculosis patients in rural areas.
To explore cigarette smoking and health-promoting behaviours among disadvantaged adults before their tuberculosis diagnosis and after their tuberculosis treatment. Although tuberculosis infection is associated with impaired immune function, healthy lifestyle habits can play a role in improving the immune system. However, limited research has explored the health-promoting behaviours and cigarette smoking habits among tuberculosis patients in Taiwan. A cross-sectional retrospective study with a convenience sample. This study was conducted between May 2013-June 2014 with 123 patients at a rural district hospital in Chiayi County, Taiwan. Statistical analyses included descriptive statistics, univariate analysis and stepwise regression analysis. Tuberculosis tended to be associated with less education, male sex, malnutrition, cigarette smoking and unhealthy lifestyle habits before the tuberculosis diagnosis. The percentage of smoking decreased from 46·9% before to 30·2% after the tuberculosis diagnosis. Body mass index and health-promoting behaviours also significantly improved after tuberculosis treatment. After controlling for potential confounding factors, multivariate analysis identified chronic disease and completed treatment as significant factors that were associated with current health-promoting behaviours. A high prevalence of cigarette smoking and low levels of health-promoting behaviours were observed before the diagnosis and during or after completing tuberculosis treatment. This study's findings indicate the importance of promoting healthy lifestyle changes among tuberculosis patients; aggressive measures should be implemented immediately after the first diagnosis of tuberculosis. Furthermore, health promotion and smoking cessation programmes should be initiated in the general population to prevent activation of latent tuberculosis infection, and these programmes should specifically target men and rural residents.


Engineering and public health at CDC.
Engineering is the application of scientific and technical knowledge to solve human problems. Using imagination, judgment, and reasoning to apply science, technology, mathematics, and practical experience, engineers develop the design, production, and operation of useful objects or processes. During the 1940s, engineers dominated the ranks of CDC scientists. In fact, the first CDC director, Assistant Surgeon General Mark Hollis, was an engineer. CDC engineers were involved in malaria control through the elimination of standing water. Eventually the CDC mission expanded to include prevention and control of dengue, typhus, and other communicable diseases. The development of chlorination, water filtration, and sewage treatment were crucial to preventing waterborne illness. Beginning in the 1950s, CDC engineers began their work to improve public health while developing the fields of environmental health, industrial hygiene, and control of air pollution. Engineering disciplines represented at CDC today include biomedical, civil, chemical, electrical, industrial, mechanical, mining, and safety engineering. Most CDC engineers are located in the National Institute for Occupational Safety and Health (NIOSH) and the Agency for Toxic Substances and Disease Registry (ATSDR). Engineering research at CDC has a broad stakeholder base. With the cooperation of industry, labor, trade associations, and other stakeholders and partners, current work includes studies of air contaminants, mining, safety, physical agents, ergonomics, and environmental hazards. Engineering solutions remain a cornerstone of the traditional "hierarchy of controls" approach to reducing public health hazards.


Seasonal Patterns of Stored-Product Insects at a Rice Mill.
The temporal and spatial patterns in flight activity outside of a rice mill were evaluated for the lesser grain borer [Rhyzopertha dominica (F.)], warehouse beetle [Trogoderma variabile Ballion], cigarette beetle [Lasioderma serricorne (F.)], and Indian meal moth [Plodia interpunctella (Hüϋbner)] to determine critical times of year when the mill would be vulnerable to invasion. Insect activity was monitored using pheromone-baited glue traps (N = 99) from June 2008 to October 2010. Traps were placed along exterior walls of all major buildings and along the fence around the perimeter of the facility. Trogoderma variabile was the most abundant species, with flight activity between mid-March and November. No activity of T. variabile was observed during December through March. Rhyzopertha dominica was also abundant, with activity in mid-April through October. A few adult R. dominica were captured in traps during winter months in the first year of study. Trap captures for all four species increased with an increase in temperature and can be described by linear equations. Knowing seasonal patterns in insect activity allows rice facilities to better understand when facilities are most vulnerable to pest activity. However, this study demonstrates that more research is needed to address how insects are immigrating and emigrating within and around a rice mill.


Insight into the Key Restriction of BiVO4 Photoanodes Prepared by Pyrolysis Method for Scalable Preparation.
Bismuth Vanadate (BiVO4 ) photoanode has been popularly investigated for promising solar water oxidation, but its intrinsic performance has been greatly retarded by the direct pyrolysis method. Here we insight the key restriction of BiVO4 prepared by metal-organic decomposition (MOD) method. It is found that the evaporation of vanadium during the pyrolysis tends to cause a substantial phase impurity, and the unexpected few tetragonal phase inhibits the charge separation evidently. Consequently, suitably excessive vanadium precursor was adopted to eliminate the phase impurity, based on which the obtained intrinsic BiVO4 photoanode could exhibit photocurrent density of 4.2 mA cm-2 at 1.23 VRHE under AM 1.5 G irradiation, as comparable to the one fabricated by the currently popular two-step electrodeposition method. Furthermore, the excellent performance can be maintained on the enlarged photoanode (25 cm2 ), demonstrating the advantage of MOD method in scalable preparation. Our work provides new insight and highlights the glorious future of MOD method for the design of scale-up efficient BiVO4 photoanode.


Prejudices and their healthcare implications. Lessons learnt from a national survey.
Introduction: The presence and recognition of prejudice during care were examined among healthcare professionals towards the Roma population, the largest ethnic minority in Hungary. Aim: The aim of this study was to explore the extent to which prejudice in the Hungarian healthcare system might affect the quality of care and, thereby, the health of the Roma patients. Method: Semi-structured interviews were conducted between June 2017 and May 2018 with 13 interviewees. Qualitative methods were applied to analyse the transcripts. Results: Our respondents generally confirmed the presence of prejudices in the healthcare system. Prejudicial attitudes and discriminatory behaviour are less common in wards where the prevalence of disadvantaged patients (HIV-positive, homeless) is high (e.g., infectology). Prejudice is not specifically directed at the Roma ethnic minority, but also extends to populations living under poor socio-economic conditions, or with underlying self-destructive behaviour (alcoholism, drug use). In the opinion of our interviewees, occasional prejudicial behaviour does not lead to sub-standard care, but, on the contrary, to positive discrimination. Prejudice does not directly lead to poorer health, but it cannot be ruled out that it may indirectly contribute to it, in the case if the patient does not seek medical attention due to a former negative experience within healthcare. Conclusion: Our results confirmed the occasional presence of prejudice against the Romas in the healthcare system, however, this may not directly contribute to Romas' poorer health status. Our results also highlighted the importance of prevention, including the education of both parties and the prevention of the burnout of care providers. Orv Hetil. 2020; 161(19): 789-796.


Automated Classification of Resting-State fMRI ICA Components Using a Deep Siamese Network.
Manual classification of functional resting state networks (RSNs) derived from Independent Component Analysis (ICA) decomposition can be labor intensive and requires expertise, particularly in large multi-subject analyses. Hence, a fully automatic algorithm that can reliably classify these RSNs is desirable. In this paper, we present a deep learning approach based on a Siamese Network to learn a discriminative feature representation for single-subject ICA component classification. Advantages of this supervised framework are that it requires relatively few training data examples and it does not require the number of ICA components to be specified. In addition, our approach permits one-shot learning, which allows generalization to new classes not seen in the training set with only one example of each new class. The proposed method is shown to out-perform traditional convolutional neural network (CNN) and template matching methods in identifying eleven subject-specific RSNs, achieving 100% accuracy on a holdout data set and over 99% accuracy on an outside data set. We also demonstrate that the method is robust to scan-rescan variation. Finally, we show that the functional connectivity of default mode and salience networks identified by the proposed technique is altered in a group analysis of mild traumatic brain injury (TBI), severe TBI, and healthy subjects.


Australian Public Attitudes on Gene Editing of the Human Embryo.
Technology is now available which facilitates gene editing and has recently been applied internationally to embryos in the laboratory. A 2002 law in Australia prohibits making heritable changes in embryos, regardless of whether the treated embryo is discarded thereafter. We sought to begin to understand public opinion in Australia about this matter, using a questionnaire given to the audience attending a Q and A panel of experts. We found majority support for allowing heritable changes for health purposes. If this is confirmed in a larger survey of the population, we suggest the existing law should be reviewed.


Differential self-assembly behaviors of cyclic and linear peptides.
Here we ask the fundamental questions about the effect of peptide topology on self-assembly. The study revealed that the self-assembling behaviors of cyclic and linear peptides are significantly different in several respects, in addition to sharing several similarities. Their clear differences included the morphological dissimilarities of the self-assembled nanostructures and their thermal stability. The similarities include their analogous critical aggregation concentration values and cytotoxicity profiles, which are in fact closely related. We believe that understanding topology-dependent self-assembly behavior of peptides is important for developing tailor-made self-assembled peptide nanostructures.


Impact of 2018 ESC/ESH and 2017 ACC/AHA Hypertension Guidelines: Difference in Prevalence of White-Coat and Masked Hypertension.
Our study evaluated whether there were differences in the prevalence of white-coat hypertension (WH) and masked hypertension (MH) based on the 2018 ESC/ESH and 2017 ACC/AHA hypertension guidelines in Korea. The motivation was the lowering of the diagnostic threshold for hypertension in the 2017 ACC/AHA guidelines. Of 319 participants without antihypertensive drug history and with suspected hypertension based on outpatient clinic blood pressure (BP) measured by physicians, 263 participants (51.6 ± 9.6 years; 125 men) who had valid research-grade office BP and 24-h ambulatory BP measurements were enrolled. WH prevalence based on daytime ambulatory BP among normotensive individuals was lower with the ESC/ESH guidelines than the ACC/AHA guidelines (29.0% vs. 71.4%, p < 0.001). However, MH prevalence based on daytime ambulatory BP among hypertensive individuals was higher based on the ESC/ESH guidelines (21.6% vs. 1.8%, p < 0.001). Seventy percent of WH cases (2017 ACC/AHA guidelines) and 95.2% of MH cases (2018 ESC/ESH guidelines) occurred in individuals with systolic BP of 130-139 mmHg and/or diastolic BP of 80-89 mmHg. The diagnostic threshold of the 2017 ACC/AHA guidelines yielded a higher prevalence of WH compared to that of the 2018 ESC/ESH guidelines. However, the prevalence of MH was higher with the 2018 ESC/ESH guidelines than with the 2017 ACC/AHA guidelines. The high prevalence of WH and MH in people with a systolic BP of 130-139 mmHg or diastolic BP of 80-89 mmHg suggests the need for a more active out-of-office BP measurement in this patient group.


Membrane Association Dictates Ligand Specificity for the Innate Immune Receptor NOD2.
The human gut must regulate its immune response to resident and pathogenic bacteria, numbering in the trillions. The peptidoglycan component of the bacterial cell wall is a dense and rigid structure that consists of polymeric carbohydrates and highly cross-linked peptides which offers protection from the host and surrounding environment. Nucleotide-binding oligomerization domain-containing protein 2 (NOD2), a human membrane-associated innate immune receptor found in the gut epithelium and mutated in an estimated 30% of Crohn's disease patients, binds to peptidoglycan fragments and initiates an immune response. Using a combination of chemical synthesis, advanced analytical assays, and protein biochemistry, we tested the binding of a variety of synthetic peptidoglycan fragments to wild-type (WT)-NOD2. Only when the protein was presented in the native membrane did binding measurements correlate with a NOD2-dependent nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) response, supporting the hypothesis that the native-membrane environment confers ligand specificity to the NOD2 receptor for NF-κB signaling. While N-acetyl-muramyl dipeptide (MDP) has been thought to be the minimal peptidoglycan fragment necessary to activate a NOD2-dependent immune response, we found that fragments with and without the dipeptide moiety are capable of binding and activating a NOD2-dependent NF-κB response, suggesting that the carbohydrate moiety of the peptidoglycan fragments is the minimal functional epitope. This work highlights the necessity of studying NOD2-ligand binding in systems that resemble the receptor's natural environment, as the cellular membrane and/or NOD2 interacting partners appear to play a crucial role in ligand binding and in triggering an innate immune response.


Differential expression of HLA class II genes associated with disease susceptibility and progression in rheumatoid arthritis.
Rheumatoid arthritis (RA)-associated HLA class II genes are assumed to promote susceptibility to and/or progression of the disease. Among the various modes of action proposed so far is the effect of the differential expression of HLA class II genes in different types of antigen-presenting cells on the Th1/Th2 balance. The aim of this study was to investigate the differential expression of genes encoded within the RA-associated HLA-DR4 superhaplotype and within the neutral DR7 and DR9 superhaplotypes. The promoters encoded within these 3 haplotypes were first analyzed for sequence polymorphisms. To test for functional consequences, we assumed that the binding of nuclear factors to the promoter elements was correlated with the transcription activity, and we used surface plasmon resonance technology. To that end, oligonucleotides representing the polymorphic regulatory sequences and nuclear extracts from a monocyte cell line and a B cell line were used. While the promoters of the highly polymorphic HLA-DRB1*04, *07, and *09 alleles showed comparable binding of nuclear factors, differential binding was observed for the 2 promoters that drive the relatively nonpolymorphic DRB4 alleles in linkage disequilibrium with DRB1. Interestingly, analysis of RA patients positive for DR4, DR7, and DR9 revealed the segregation of radiographic progression with the stronger of the 2 DRB4 promoters, independent of the DRB1 allele. Moreover, DRB1*04 alleles in RA patients showed a reduced association with the DRB4 splice variant, completely preventing DRB4 expression. Our findings represent the first evidence of a correlation between the differential expression of HLA class II genes and both the susceptibility and the progression of RA.


Degradation Kinetics of Clavulanic Acid in Fermentation Broths at Low Temperatures.
Clavulanic acid (CA) is a β-lactam antibiotic inhibitor of β-lactamase enzymes, which confers resistance to bacteria against several antibiotics. CA is produced in submerged cultures by the filamentous Gram-positive bacterium Streptomyces clavuligerus; yield and downstream process are compromised by a degradation phenomenon, which is not yet completely elucidated. In this contribution, a study of degradation kinetics of CA at low temperatures (-80, -20, 4, and 25 °C) and pH 6.8 in chemically-defined fermentation broths is presented. Samples of CA in the fermentation broths showed a fast decline of concentration during the first 5 h followed by a slower, but stable, reaction rate in the subsequent hours. A reversible-irreversible kinetic model was applied to explain the degradation rate of CA, its dependence on temperature and concentration. Kinetic parameters for the equilibrium and irreversible reactions were calculated and the proposed kinetic model was validated with experimental data of CA degradation ranging 16.3 mg/L to 127.0 mg/L. Degradation of the chromophore CA-imidazole, which is commonly used for quantifications by High Performance Liquid Chromatography, was also studied at 4 °C and 25 °C, showing a rapid rate of degradation according to irreversible first-order kinetics. A hydrolysis reaction mechanism is proposed as the cause of CA-imidazole loss in aqueous solutions.


Treating black hypertensives with capozide.
Twenty-four black men with mild to moderate essential hypertension were enrolled in an open-label trial comparing the efficacy of two doses of Capozide (captopril and hydrochlorothiazide). All antihypertensive drugs were discontinued and patients then received placebo for 2 weeks. Twenty-two patients, mean age 59.1 +/- 14.3 years, with sitting diastolic blood pressure (BP) 92 to 110 mm Hg, entered the 6-week active-drug phase. Eleven patients (Group A) were randomized to Capozide 25/15 and 11 (Group B) to Capozide 50/15. Baseline mean BPs were 151.0/100.7 mm Hg in Group A and 153.1/100.7 mm Hg in Group B. At week 6, mean BPs were 128.7/84.4 mm Hg in Group A and 126.8/82.7 mm Hg in Group B. Uric acid, blood urea nitrogen and creatinine levels rose slightly in both groups. There were no adverse events. Eighteen patients had normal BPs at study completion. Twice-daily Capozide treatment is effective and well tolerated in blacks; patients responded equally well to both doses.


Aflatoxin B1 inhibition in Aspergillus flavus by Aspergillus niger through down-regulating expression of major biosynthetic genes and AFB1 degradation by atoxigenic A. flavus.
Twenty Aspergillus niger strains were isolated from peanuts and 14 strains were able to completely inhibit AFB1 production with co-cultivation. By using a Spin-X centrifuge system, it was confirmed that there are some soluble signal molecules or antibiotics involved in the inhibition by A. niger, although they are absent during the initial 24h of A. flavus growth when it is sensitive to inhibition. In A. flavus, 19 of 20 aflatoxin biosynthetic genes were down-regulated by A. niger. Importantly, the expression of aflS was significantly down-regulated, resulting in a reduction of AflS/AflR ratio. The results suggest that A. niger could directly inhibit AFB1 biosynthesis through reducing the abundance of aflS to aflR mRNAs. Interestingly, atoxigenic A. flavus JZ2 and GZ15 effectively degrade AFB1. Two new metabolites were identified and the key toxic lactone and furofuran rings both were destroyed and hydrogenated, meaning that lactonase and reductase might be involved in the degradation process.


Which model better fits the role of aire in the establishment of self-tolerance: the transcription model or the maturation model?
The discovery of Aire-dependent transcriptional control of many tissue-restricted self-antigen (TRA) genes in thymic epithelial cells in the medulla (medullary thymic epithelial cells, mTECs) has raised the intriguing question of how the single Aire gene can influence the transcription of such a large number of TRA genes within mTECs. From a mechanistic viewpoint, there are two possible models to explain the function of Aire in this action. In the first model, TRAs are considered to be the direct target genes of Aire's transcriptional activity. In this scenario, the lack of Aire protein within cells would result in the defective TRA gene expression, while the maturation program of mTECs would be unaffected in principle. The second model hypothesizes that Aire is necessary for the maturation program of mTECs. In this case, we assume that the mTEC compartment does not mature normally in the absence of Aire. If acquisition of the properties of TRA gene expression depends on the maturation status of mTECs, a defect of such an Aire-dependent maturation program in Aire-deficient mTECs can also result in impaired TRA gene expression. In this brief review, we will focus on these two contrasting models for the roles of Aire in controlling the expression of TRAs within mTECs.


Pulsed electromagnetic fields accelerate apoptotic rate in osteoclasts.
Selective control of cell function by applying specifically configured, low-energy, time-varying electromagnetic fields (EMF) has added a new, exciting dimension to biology and medicine. However, the mechanism involved is less clear. In our study, we investigated the effect of pulsed electromagnetic fields (PEMF) on induction of osteoclasts apoptosis. A statistically significant increase of apoptotic rate in osteoclasts (48 hr after isolation) was found when exposed to 7.5 Hz PEMF with induced electric fields intensity of 3.0 muv/cm for 8 (105%, p < 0.001) and 16 hr (30%, p < 0.05). However, exposure of osteoclasts to PEMF for only 1 hr showed no statistically significant differences. These findings suggest that PEMF have the ability to speed up apoptosis of osteoclasts derived from primary osteoblasts and bone marrow cells cocultures. This in vitro study, therefore, could be considered as groundwork for in vivo PEMF applications on some osteoclasts-associated bone diseases such as osteoporosis.