Citrus fruit flavonoids influence on neutrophil apoptosis and oxidative metabolism.
Citrus flavonoids are dietary antioxidants that may protect against oxidative stress linked to inflammation and reduce the risk of macromolecule damage caused by free radicals. Three citrus flavonoids - naringin, naringenin and hesperidin - have been examined for their ability to activate caspase-3, a marker of apoptosis execution, in human polymorphonuclear neutrophils in vitro, stimulated and non-stimulated with phorbol 12-myristate 13-acetate. The flavonoid potency to reduce reactive oxygen species generation was also assessed in measurements of superoxide radical and luminol dependent chemiluminescence. The caspase-3 activity depended on the time of flavonoid treatment and was increased markedly in phorbol-treated cells incubated with flavonoids for 24 h. Hesperidin at a concentration of 100 microm exerted the strongest stimulating effect on the enzyme (137% of control). Flavonoids inhibited the neutrophil ability to generate superoxide radical and 10-100 microM hesperidin appeared the most active phytochemical. The antioxidant capacity of the examined flavonoids was most prominently expressed in the chemiluminescent measurements. The obtained results suggest that reactive oxygen species may inhibit apoptosis via caspase-3 inhibition and the antioxidant action of citrus flavonoids may reverse this process.


Turgor-responsive gene transcription and RNA levels increase rapidly when pea shoots are wilted. Sequence and expression of three inducible genes.
Reduction of turgor in pea shoots caused the accumulation of several poly(A) RNAs. cDNA clones derived from three different poly(A) RNAs which accumulate in wilted pea shoots were isolated, sequenced and expression of the corresponding genes examined. Clone 7a encoded a 289 amino acid protein. The C-terminal 180 amino acids of this protein were homologous to soybean nodulin-26. RNA hybridizing to cDNA 7a was abundant in roots, and induced in shoots by dehydration, heat shock and to a small extent by ABA. Hydropathic plots indicate that the protein encoded by cDNA 7a contains six potential membrane spanning domains similar to proteins which form ion channels. Clone 15a encoded a 363 amino acid protein with high homology to cysteine proteases. RNA hybridizing to cDNA 15a was more abundant in roots than shoots of control plants. Dehydration of pea shoots induced cDNA 15a mRNA levels whereas heat shock or ABA treatment did not. Clone 26g encoded a 508 amino acid protein with 30% residue identity to several aldehyde dehydrogenases. RNA hybridizing to cDNA 26g was induced by dehydration of shoots but not roots and heat shock and ABA did not modulate RNA levels. Levels of the three poly(A) RNAs increased 4-6-fold by 4 h after wilting and this increase was not altered by pretreatment of shoots with cycloheximide. When wilted shoots were rehydrated, RNA hybridizing to cDNA 26g declined to pre-stress levels within 2 h. Run-on transcription experiments using nuclei from pea shoots showed that transcription of the genes which encode the three poly(A) RNAs was induced within 30 min following reduction of shoot turgor. One of the genes showed a further increase in transcription by 4 h after dehydration whereas transcription of the other 2 genes declined. These results indicate that plant cells respond to changes in cell turgor by rapidly increasing transcription of several genes. Furthermore, the expression of the turgor-responsive genes varies with respect to the time course of induction and reversibility of the wilting-induced changes.


Third-trimester amniotic fluid metal levels associated with preeclampsia.
Studies of maternal amniotic fluid and serum at delivery have revealed the presence of toxic metals or deficiencies of essential metals associated with high-risk pregnancy. Essential and toxic metal levels were studied in 29 preeclamptic and 101 normal pregnancies. The authors grouped the samples according to the following gestation ages: 33-36 wk (48 normal and 10 preeclamptic) and 37-40 wk (53 normal and 19 preeclamptic). The metals studied were calcium, magnesium, zinc, selenium, copper, cadmium, and lead. Comparisons of the 33-36-wk gestation group showed significant differences between normal and preeclamptic amniotic fluid in levels of lead, calcium, magnesium, zinc, and selenium. There were also significant differences in levels of magnesium, copper, zinc, cadmium, and lead during the gestation period of 37-40 wk. The changes observed in calcium and lead levels were consistent with the results of animal studies in which investigators have found depressed calcium transport associated with subacute or chronic lead poisoning, resulting in a physical syndrome similar to preeclampsia.


[Dapoxetine in treatment of premature ejaculation: a systematic review].
To assess the effectiveness of dapoxetine in the treatment of premature ejaculation. Both English and Chinese studies involving men with prematrue ejaculation who were treated with dapoxetine from the Cochrane Library, MEDLINE, EMBASE and CNKI, CBM, VIP between 1979 and 2009.were included in the randomized controlled trials (RCTs) and the data processed by RevMan. Five RCTs involving 4433 patients were included in the Meta analysis, of which 3 were of grade A and 2 were of grade B according to the quality evaluation of methodology. Intravaginal ejaculatory latency time (IELT), patient-reported global impression of change (PGI), satisfaction with sexual intercourse (SWSI), perceived control over ejaculation (PCOE), personal distress related to ejaculation (PDRE) were used for assessment. Meta analysis based on included studies of patients having been treated with dapoxetine for 9-24 weeks showed that: (1) the difference of the patients' IELT between treatment group and control group was statistically significant [P<0.001, WMD (95%CI) was 1.38 (1.21,1.55)]; (2) the difference of the PGI of development in premature ejaculation between treatment group and control group was statistically significant [P<0.001, OR (95%CI) was 3.56 (2.60,4.88)]; (3) the difference of the patients' SWSI between treatment group and control group was statistically significant [P<0.001, OR (95%CI) was 3.85 (2.08,7.10)]; the difference of the patients' score of SWSI between treatment group and control group was statistically significant [P<0.001, WMD (95%CI) was 0.55 (0.48,0.62)]; (4) the difference of the patients' change of PCOE between treatment group and control group was statistically significant [P<0.001, OR (95%CI) was 2.87(2.30,3.58)]; the difference of the patients' score of PCOE between treatment group and control group was statistically significant [P<0.001, WMD (95%CI) was 0.63(0.49,0.78)]; (5) after being treated with dapoxetine for 9-24 weeks, the difference of the patients' change of PDRE between treatment group and control group was statistically significant [P<0.001, OR (95%CI) was 2.02 (1.69,2.42)]. All the RCTs reported the side effects of dapoxetine, but the results showed that there were no serious side effects of dapoxetine during the treatment period. The available evidence indicates that dapoxetine would improve the symptoms of premature ejaculation, prolong IELT over 9-24 weeks in men from a wide range of cultural backgrounds, and significantly improve all patients' reported outcomes and the patients' clinical global impressions of premature ejaculation, including more control, greater satisfaction, and less distress.


Malignant tumor formation in dogs previously irradiated for acanthomatous epulis.
In this retrospective study of 57 dogs irradiated for oral acanthomatous epulis, 2 (3.5%) dogs developed a second tumor (sarcoma, osteosarcoma) in the radiation treatment field at 5.2 and 8.7 years after the end of radiation therapy. As opposed to previous reports, no second epithelial tumors developed in the radiation treatment field. There is a risk of radiation-induced carcinogenesis, but it appears that it is a relatively low risk and an event that occurs years after radiation therapy. Radiation-induced tumors are of more concern in younger dogs that undergo radiation therapy for tumors that are radioresponsive, such as acanthomatous epulis, where long-term survival is expected. The only statistically significant variable in the survival analysis was age, with dogs less than 8.3 years old having a significantly longer median overall survival (2322 days) than dogs older than 8.3 years (1106 days; P<0.0001).


[Sub-field imaging spectrometer design based on Offner structure].
To satisfy imaging spectrometers's miniaturization, lightweight and large field requirements in space application, the current optical design of imaging spectrometer with Offner structure was analyzed, and an simple method to design imaging spectrometer with concave grating based on current ways was given. Using the method offered, the sub-field imaging spectrometer with 400 km altitude, 0.4-1.0 microm wavelength range, 5 F-number of 720 mm focal length and 4.3 degrees total field was designed. Optical fiber was used to transfer the image in telescope's focal plane to three slits arranged in the same plane so as to achieve subfield. The CCD detector with 1 024 x 1 024 and 18 microm x 18 microm was used to receive the image of the three slits after dispersing. Using ZEMAX software optimization and tolerance analysis, the system can satisfy 5 nm spectrum resolution and 5 m field resolution, and the MTF is over 0.62 with 28 lp x mm(-1). The field of the system is almost 3 times that of similar instruments used in space probe.


Peripheral nerve regeneration with conduits: use of vein tubes.
Treatment of peripheral nerve injuries remains a challenge to modern medicine due to the complexity of the neurobiological nerve regenerating process. There is a greater challenge when the transected nerve ends are not amenable to primary end-to-end tensionless neurorraphy. When facing a segmental nerve defect, great effort has been made to develop an alternative to the autologous nerve graft in order to circumvent morbidity at donor site, such as neuroma formation, scarring and permanent loss of function. Tubolization techniques have been developed to bridge nerve gaps and have been extensively studied in numerous experimental and clinical trials. The use of a conduit intends to act as a vehicle for moderation and modulation of the cellular and molecular ambience for nerve regeneration. Among several conduits, vein tubes were validated for clinical application with improving outcomes over the years. This article aims to address the investigation and treatment of segmental nerve injury and draw the current panorama on the use of vein tubes as an autogenous nerve conduit.


[Carotid body tumour. Case report and literature review].
The tumour of the carotid body is rare. About 1000 cases had been reported in the literature. It may occur sporadically in 90% of cases and it affects both sexes in the same proportion and in the middle age. This tumour may be misdiagnosed if it is not suspected. Ultrasono-graphy and color-Doppler scan show a hypervascular tumour between the internal and external carotid arteries. CT-scan defines the tumour s extent on the surrounding structures. Angiography is the gold standard for diagnosis, showing a hypervascular mass displacing the bifurcation of the carotid arteries. Sometimes radiotherapy and embolization are indicated but the surgical excision of carotid body tumours is the therapy of choice. The surgical approach through incision like carotid artery operation is performed. If the subadventitial plane between tumour and arterial wall is not identified, resection of carotid artery and insertion of a shunt is required. Although the diagnosis and the surgical technique advances, the incidence of postoperative nerve injury is high in the different series. The clinical suspect and the early diagnosis are very important because low morbidity rate occurs with resection of a small chemodectoma. The surgical excision can be followed by postoperative respiratory depression or dyspnea both with regional and general anesthesia. The authors report a case of a medium size tumour operated on and developing a mild transient weakness of cranial nerve VII. Recent trends in evaluation and therapy are analysed and the literature is reviewed.


[Mid-term effectiveness of arthroscopic anterior cruciate ligament reconstruction combined with meniscus allograft transplantation].
To summarize the mid-term effectiveness of arthroscopic anterior cruciate ligament (ACL) reconstruction combined with meniscus allograft transplantation. A clinical data of 21 patients treated with arthroscopic ACL reconstruction and meniscus allograft transplantation and followed up more than 5 years between February 2007 and December 2014 was retrospectively analyzed. There were 12 males and 9 females, aged from 18 to 45 years, with an average age of 23.5 years. The cause of injury was sport sprain in 15 cases, falling in 4 cases, and traffic accident in 2 cases. The time from injury to operation ranged from 2 to 36 months, with an average of 12 months. Among them, 15 patients underwent previous meniscectomy, with an average interval of 1.6 years (range, 3 months to 6.5 years). All patients were primary ACL reconstruction. Preoperative anterior drawer test, Lachman test, and pivot shift test were positive. Lysholm score was 43.6±10.2. International Knee Documentation Committee (IKDC) score was 60.50±14.06. Of the 21 patients, 10 were gradeⅠ-Ⅱcartilage injuries and 11 were grade Ⅲ cartilage injuries according to MRI. All patients were followed up 5.1-7.8 years, with an average of 5.5 years. There were 2 cases of numbness of lower extremity, 3 cases of slight exudation of incision, 2 cases of articular movement bounce, 5 cases of mild joint swelling and pain after exercise. At last follow-up, Lachman tests were negative in 18 cases and positive in 3 cases; anterior drawer tests were negative in 19 cases and positive in 2 cases; pivot shift tests were negative in all cases. Lysholm score was 84.5±16.5 and IKDC score was 85.25±4.60, which were significantly higher than those before operation ( P<0.01). The flexion and extension of the affected knee joint were (128±13) and (3±7)°, respectively, which were smaller than those of the healthy knee joint [(133±15), (0±5)°] ( P<0.01). The results of KT-1000 test showed that when knee flexion was 30 and 90°, tibial anterior displacement of affected side [(2.35±1.20), (1.60±1.15) mm] were not significantly different from those of healthy side [(1.20±1.10), (1.10±1.03) mm] ( P>0.01). MRI showed that the ACL graft was in normal position and meniscus survived well. Cartilage injuries were gradeⅠ-Ⅱ in 18 cases and grade Ⅲ in 3 cases. For patients with severe meniscus injury and ACL rupture, ACL reconstruction combined with meniscus allograft transplantation can restore the stability of the joint, recover the meniscus function which is conducive to the protection of articular cartilage and obtain satisfactory mid-term effectiveness.


Estimating the cost of cervical cancer screening in five developing countries.
Cost-effectiveness analyses (CEAs) can provide useful information to policymakers concerned with the broad allocation of resources as well as to local decision makers choosing between different options for reducing the burden from a single disease. For the latter, it is important to use country-specific data when possible and to represent cost differences between countries that might make one strategy more or less attractive than another strategy locally. As part of a CEA of cervical cancer screening in five developing countries, we supplemented limited primary cost data by developing other estimation techniques for direct medical and non-medical costs associated with alternative screening approaches using one of three initial screening tests: simple visual screening, HPV DNA testing, and cervical cytology. Here, we report estimation methods and results for three cost areas in which data were lacking. To supplement direct medical costs, including staff, supplies, and equipment depreciation using country-specific data, we used alternative techniques to quantify cervical cytology and HPV DNA laboratory sample processing costs. We used a detailed quantity and price approach whose face validity was compared to an adaptation of a US laboratory estimation methodology. This methodology was also used to project annual sample processing capacities for each laboratory type. The cost of sample transport from the clinic to the laboratory was estimated using spatial models. A plausible range of the cost of patient time spent seeking and receiving screening was estimated using only formal sector employment and wages as well as using both formal and informal sector participation and country-specific minimum wages. Data sources included primary data from country-specific studies, international databases, international prices, and expert opinion. Costs were standardized to year 2000 international dollars using inflation adjustment and purchasing power parity. Cervical cytology laboratory processing costs were I$1.57-3.37 using the quantity and price method compared to I$1.58-3.02 from the face validation method. HPV DNA processing costs were I$6.07-6.59. Rural laboratory transport costs for cytology were I$0.12-0.64 and I$0.14-0.74 for HPV DNA laboratories. Under assumptions of lower resource efficiency, these estimates increased to I$0.42-0.83 and I$0.54-1.06. Estimates of the value of an hour of patient time using only formal sector participation were I$0.07-4.16, increasing to I$0.30-4.80 when informal and unpaid labor was also included. The value of patient time for traveling, waiting, and attending a screening visit was I$0.68-17.74. With the total cost of screening for cytology and HPV DNA testing ranging from I$4.85-40.54 and I$11.30-48.77 respectively, the cost of the laboratory transport, processing, and patient time accounted for 26-66% and 33-65% of the total costs. From a payer perspective, laboratory transport and processing accounted for 18-48% and 25-60% of total direct medical costs of I$4.11-19.96 and I$10.57-28.18 respectively. Cost estimates of laboratory processing, sample transport, and patient time account for a significant proportion of total cervical cancer screening costs in five developing countries and provide important inputs for CEAs of alternative screening modalities.


Elevated relative fluorescence intensity of CD38 antigen expression on CD8+ T cells is a marker of poor prognosis in HIV infection: results of 6 years of follow-up.
Relative fluorescence intensity measurements from a flow cytometer were used to evaluate expression of CD38 and HLA-DR antigens. These molecules are associated with cellular activation and are present at increased levels on the CD8+ lymphocytes of HIV-infected subjects. In the current study, the prognostic value of mean fluorescence intensity measurements of CD38 and HLA-DR on CD8+ cells was compared to results from our previous study in which we reported prognostic value for an elevated percentage of CD8+ cells that were positive for expression of the CD38 antigen (Giorgi et al.: JAIDS 6:904-912, 1993). Using the proportional hazards model, elevated mean fluorescence intensity of CD38 expression on CD8+ cells had prognostic value for development of AIDS that was almost identical to the prognostic value of the percentage of CD8+ cells that were positive for expression of CD38. This prognostic value was in addition to that provided by the patient's CD4+ cell measurement. To our knowledge, this is the first report that a measurement of fluorescence intensity can be used as a prognostic marker in an immunodeficiency disease. Efforts are needed to establish methods that will allow widespread application of this observation in the clinical management of HIV-infected subjects.


[The influence of meteorological factors for frequency of Herpes simplex keratitis].
The variation of the meteorological factors could be a stress for the organism, which could determine the start of some diseases. We studied 252 cases of herpes simplex keratitis which had been hospitalized in Eye Clinic of Cluj between 1983-1991. The selection criteria were: known diagnosis; day of the rise could be exactly established; disease was taken place in Cluj or in region not longer than 30 km on Somes Valley. The meteorological parameters were collected by specialist from the Meteorological Station of the city and from the Cluj's Airport. The results were verified statistical and were guaranted by p = 0.05. The herpes simplex keratitis are more frequently in days with a temperature higher and an atmospheric pressure lower than the average value of the season. We drew a possible pathogenic mechanism to explained the reactivation of the virus under the influence of warm weather.


[Differential inhibition of raubasine and tetrahydroalstonine on the vasopressor response to sympathetic nervous stimulation and intravenous noradrenaline in the pithed rat].
The effects of raubasine and tetrahydroalstonine (THA) on the vasopressor response to sympathetic nerve stimulation and to i.v. administration of noradrenaline were studied in the pithed rat to ascertain whether raubasine and THA would preferentially block pressor responses due to exogenous versus endogenous noradrenaline alpha-adrenergic receptor activation. Raubasine (0.5 to 4 mg/kg i.v.) was more effective in blocking the response due to sympathetic nerve stimulation than that due to i.v. noradrenaline. On the other hand THA (0.5 to 4 mg/kg i.v.) produced greater inhibition of the i.v. noradrenaline response than the sympathetic nerve stimulation response. THA (0.5, 1, 2 mg/kg i.v.) enhanced the nerve stimulation response, while at the 4 mg/kg dose the response was slightly reduced. This may be explained by a preferential block by THA at low doses, of presynaptic alpha 2-adrenoceptors. In pithed rat raubasine and THA showed dose-related blocking effects on the pressor response of phenylephrine and B-HT 933, respectively. This suggest that raubasine preferentially blocks alpha-1 and THA alpha 2-adrenoceptors. The results suggest that raubasine and THA preferentially block the pressor responses of post-synaptic alpha-adrenergic receptor activation due to endogenous and exogenous noradrenaline, respectively.


High resolution electron tomography and segmentation-by-modeling interpretation in Bsoft.
Bsoft offers many tools for the processing of tomographic tilt series and the interpretation of tomograms. Since I introduced tomography into Bsoft almost two decades ago, the field has advanced significantly, requiring refinement of old algorithms and development of new ones. The current direct detectors allow us to collect data more efficiently and with better quality, progressing towards automation. The goal is then to also automate alignment of tilt series and reconstruction. I added an estimation of the specimen thickness as well as fiducialless alignment, to augment the existing fiducial-based alignment. High-resolution work requires correction for the contrast transfer function, in tomography complicated by the tilted specimen. For this, I developed a method to generate a power spectrum using the whole micrograph, compensating for tilting. This is followed by routine determination of the contrast transfer function, and correction for it during reconstruction. The next steps involve interpretation of the tomogram, either by subtomogram averaging where possible, or by segmentation and modeling otherwise. Such interpretation actually constitutes the main time-consuming part of tomography and is less amenable to automation compared to the initial reconstruction.


Neuromuscular Patients Are 40% More Likely to Get a Cast Injury.
Cast injuries can occur during application, throughout immobilization, and during removal, with common morbidities being pressure ulcers and cast saw burns. The incidence rate of cast injuries in generalized and diagnosis specific pediatric populations is not known. The goal of this study is to accurately quantify the rate of incidence of cast injuries at a large pediatric orthopaedic practice and identify potentially modifiable risk factors to guide quality of care improvement. A retrospective review was performed at our institution between July 2019 and October 2020. Inclusion criteria was all pediatric patients (below 21 y old) with an orthopaedic diagnosis that was treated with casting. The child's diagnosis, specific type of cast, and training level of the person applying the cast was recorded. Patient injuries were identified through cast technicians' documentation regarding cast removal. The primary outcome was the incidence of casting injuries from July 2019 through October 2020. Additional outcomes included the association between diagnosis of neuromuscular disease and training level of individual applying the cast with casting injuries. χ2 tests were used to compare categorical variables and post hoc comparisons using Bonferroni correction. Injury incidence rates were calculated as number of injuries per 1000 casts. There were 2239 casts placed on children at this institution between July 2019 and October 2020 and a total of 28 injuries for an incidence rate of 12.5 per 1000. Of the 28 total injuries reported, there were 5 cast saw burns (2.2 per 1000) and 23 pressure ulcers (10.3 per 1000). Incidence of cast injury was not significantly correlated with timing of application during the academic year or training level of the individual applying the cast (P=0.21 and 0.86). Notably, there was a significantly higher incidence of cast injuries in individuals with a diagnosis of a neuromuscular disorder (37.4 per 1000) than those without (7.5 per 1000) (P<0.01). The incidence of cast injuries is 12.5 per 1000 children at our level I trauma tertiary referral pediatric clinic. Training level of the individual applying the cast or timing during the academic year did not correlate with cast injuries. Patients with neuromuscular disorders are at significantly higher risk for experiencing cast injuries. Level III.


Codelivery of doxorubicin and elacridar to target both liver cancer cells and stem cells by polylactide-co-glycolide/d-alpha-tocopherol polyethylene glycol 1000 succinate nanoparticles.
Liver cancer is the third leading cause of cancer-related deaths worldwide. Liver cancer stem cells (LCSCs) are a subpopulation of cancer cells that are responsible for the initiation, progression, drug resistance, recurrence, and metastasis of liver cancer. Recent studies have suggested that the eradication of both LCSCs and liver cancer cells is necessary because the conversion of cancer stem cells (CSCs) to cancer cells occasionally occurs. As ATP-binding cassette (ABC) transporters are overexpressed in both CSCs and cancer cells, combined therapies using ABC transporter inhibitors and chemotherapy drugs could show superior therapeutic efficacy in liver cancer. In this study, we developed poly(lactide-co-glycolide)/d-alpha-tocopherol polyethylene glycol 1000 succinate nanoparticles to accomplish the simultaneous delivery of an optimized ratio of doxorubicin (DOX) and elacridar (ELC) to target both LCSCs and liver cancer cells. Median-effect analysis was used for screening of DOX and ELC for synergy in liver cancer cells (HepG2 cells) and LCSCs (HepG2 tumor sphere [HepG2-TS]). Then, nanoparticles loaded with DOX and ELC at the optimized ratio (NDEs) were prepared by nanoprecipitation method. The cytotoxicity and colony and tumor sphere formation ability of nanoparticles were investigated in vitro, and the tissue distribution and antitumor activity of nanoparticles were evaluated in vivo. We demonstrated that a DOX/ELC molar ratio of 1:1 was synergistic in HepG2 cells and HepG2-TS. NDEs were shown to exhibit significantly increased cytotoxic effects against both HepG2 and HepG2-TS compared with DOX-loaded nanoparticles (NDs) or ELC-loaded nanoparticles (NEs) in vitro. In vivo studies demonstrated that the nanoparticles exhibited better tumor targeting, with NDE showing the strongest antitumor activity with lower systemic toxicity. These results suggested that NDE represented a promising combination therapy against liver cancer by targeting both liver cancer cells and CSCs.


Adiponectin and chronic kidney disease; a review on recent findings.
Adiponectin is a multifunctional cytokine that has a role in regulating inflammation. In patients without chronic renal failure (CRF) and type 2 diabetics, decreased adiponectin levels are associated with insulin resistance. Lower serum plasma adiponectin values are link to larger tumor size and metastasis in clear-cell carcinoma of the kidney too. However, in patients with established chronic kidney disease (CKD), adiponectin levels are elevated and positively predict progression of disease. In addition, increased levels of serum adiponectin of hemodialysis patients were associated with decrease in bone mineral density in hemodialysis patients. Thus, depending on type of renal failure should be adjusted the adiponectin levels in patients. In CKD patients without diabetic, decreasing adiponectin levels by ARB drugs may be appropriate for inhibition of disease progression.


Activation of Munc13-1 by Diacylglycerol (DAG)-Lactones.
Munc13-1 is a key protein necessary for vesicle fusion and neurotransmitter release in the brain. Diacylglycerol (DAG)/phorbol ester binds to its C1 domain in the plasma membrane and activates it. The C1 domain of Munc13-1 and protein kinase C (PKC) are homologous in terms of sequence and structure. In order to identify small-molecule modulators of Munc13-1 targeting the C1 domain, we studied the effect of three DAG-lactones, (R,Z)-(2-(hydroxymethyl)-4-(3-isobutyl-5-methylhexylidene)-5-oxotetrahydrofuran-2-yl)methyl pivalate (JH-131e-153), (E)-(2-(hydroxymethyl)-4-(3-isobutyl-5-methylhexylidene)-5-oxotetrahydrofuran-2-yl)methyl pivalate (AJH-836), and (E)-(2-(hydroxymethyl)-4-(4-nitrobenzylidene)-5-oxotetrahydrofuran-2-yl)methyl 4-(dimethylamino)benzoate (130C037), on Munc13-1 activation using the ligand-induced membrane translocation assay. JH-131e-153 showed higher activation than AJH-836, and 130C037 was not able to activate Munc13-1. To understand the role of the ligand-binding site residues in the activation process, three alanine mutants were generated. For AJH-836, the order of activation was wild-type (WT) Munc13-1 > R592A > W588A > I590A. For JH-131e-153, the order of activation was WT > I590 ≈ R592A ≈ W588A. Overall, the Z isomer of DAG-lactones showed higher potency than the E isomer and Trp-588, Ile-590, and Arg-592 were important for its binding. When comparing the activation of Munc13-1 and PKC, the order of activation for JH-131e-153 was PKCα > Munc13-1 > PKCε and for AJH-836, the order of activation was PKCε > PKCα > Munc13-1. Molecular docking supported higher binding of JH-131e-153 than AJH-836 with the Munc13-1 C1 domain. Our results suggest that DAG-lactones have the potential to modulate neuronal processes via Munc13-1 and can be further developed for therapeutic intervention for neurodegenerative diseases.


Diagnosis and surgical treatment of small papillary carcinomas of the thyroid gland.
The preoperative and pathological findings of 38 cases of small papillary carcinoma of the thyroid gland measuring < or = 10 mm in diameter and 74 cases of large papillary carcinoma measuring more than 10 mm were compared. In the small carcinoma cases, the sensitivity of palpation, ultrasonography, and fine-needle aspiration biopsy (FNAB) was higher than that of computed tomography and soft tissue roentgenography of the neck. Therefore, palpation, US, and FNAB should be instituted as reliable diagnostic methods, but other examinations seem unsuitable as methods for preoperative qualitative diagnosis. No definite correlation was found between the intraglandular dissemination and the size of the carcinoma, and the frequency of metastasis was relatively high even in the small carcinoma cases. If the patient elects to undergo surgery, subtotal thyroidectomy with lymph node dissection should be performed for small thyroid carcinomas as well as large carcinomas.


[French epidemiological survey of burns in children under 5 years of age].
To add to epidemiological data concerning serious accidents in the home in France. This prospective study, carried out over 12 months, included all children with burns admitted to 14 burn units (BU) and 18 pediatric surgery units (PS) using a questionnaire from the SFETB comprising 20 items. On population 0 to 60 months, 687 patients were included (434 in BU, 253 in PS). Descriptive analysis and correlation of several parameters identified patterns of burns in childhood and the factors of severity. Victim data: young boys (59.3%), aged < 36 months (mean = 24 months), burned at home, in the kitchen (62.4%) with hot fluids (73%), or bathroom (16.2%) with tap water by immersion. Flames were cause of burns (8.7%) by inflammable products. Lesions were always more severe in surface and risk of mortality was higher. Burns covered a surface equal or less than 10% in 57% of cases. The value of the study was to be more aware of the dangers of which children are exposed and to encourage application of preventive measures.


Wilkie syndrome. What is this?
Superior mesenteric artery syndrome is extremely rare and is characterized by postprandial epigastric pain, nausea, vomiting and loss of appetite, with subsequent weight loss, which aggravates the condition of the patients. The syndrome is caused by compression of the third part of the duodenum in the angle between the aorta and the superior mesenteric artery. This review updates etiology, epidemiology, diagnosis, treatment and outcome of the superior mesenteric artery syndrome. Superior mesenteric artery syndrome is clearly defined and frequently associated with a wide range of predisposing conditions and surgical procedures.


Selectins ligand decorated drug carriers for activated endothelial cell targeting.
New active particulate polymeric vectors based on branched polyester copolymers of hydroxy-acid and allyl glycidyl ether were developed to target drugs to the inflammatory endothelial cell surface. The hydroxyl and carboxyl derivatives of these polymers allow grafting of ligand molecules on the polyester backbones at different densities. A known potent nonselective selectin ligand was selected and synthesized using a new scheme. This synthesis allowed the grafting of the ligand to the polyester polymers, preserving its binding activity as assessed by docking simulations. Selectin expression on human umbilical cord vascular endothelial cells (HUVEC) was induced with the pro-inflammatory bacterial lipopolysaccharide (LPS) or with the nonselective inhibitor of nitric oxide synthase L-NAME. Strong adhesion of the ligand decorated nanoparticles was evidenced in vitro on activated HUVEC. Binding of nanoparticles bearing ligand molecules could be efficiently inhibited by prior incubation of cells with free ligand, demonstrating that adhesion of the nanoparticles is mediated by specific interaction between the ligand and the selectin receptors. These nanoparticles could be used for specific drug delivery to the activated vascular endothelium, suggesting their application in the treatment of diseases with an inflammatory component such as rheumatoid arthritis and cancer.


Development and characterization of cIEF-MALDI-TOF MS for protein analysis.
This paper describes the hyphenation of cIEF and MALDI-TOF MS via a fractionation or spotting device. After focusing in cIEF the compounds are hydrodynamically mobilized and deposited on a MALDI target plate using a sheath liquid interface, which provides the catholyte solution and the electrical ground. From previous experiments, sample conditions that resulted in a high resolution in cIEF and acceptable protein signal intensity in MS were selected [Silvertand et al., Electrophoresis, 2008, 29, 1985-1996]. Besides the mixture of test proteins, the sample solution contains 1% Pharmalyte, 0.3% hydroxyethyl cellulose and 0.1% Tween 20 and is used for both optimization as well as characterization of the cIEF-MALDI-TOF MS system. Hyphenation problems encountered are mainly due to transfer of the liquid from the needle to the MALDI target plate and are solved by choosing the proper sheath catholyte (200 mM NH4OH in 50% methanol with 0.1% Tween20). MS electropherograms were reconstructed by plotting the intensities of the m/z values corresponding to the proteins versus migration time (related to spot number). Reproducibility, peak width and signal intensity for different focusing and spotting (fractionation) times were calculated using these reconstructed MS electropherograms as well as the UV electropherograms. The best results were obtained with focusing time of 75 min (no under- or overfocusing) and a spotting time of 5 s (highest protein signal intensity in MS). The applicability of the system is demonstrated by the analysis of a biopharmaceutical (glucagon) and its deamidation product.


Code of Silence: Students' Perceptions of School Climate and Willingness to Intervene in a Peer's Dangerous Plan.
The current study presented 1,933 adolescents from 13 schools with a scenario about a hypothetical peer's plan to "do something dangerous" at school and asked how likely they would be to respond with four different actions: intervene directly, tell a teacher or principal, discuss it with a friend but not an adult, and do nothing. High school students were less likely than those in middle school to say they would approach the peer directly or confide in a teacher or principal. Students were most likely to favor taking action on their own over all of the other response strategies. Students with positive perceptions of their schools were more likely to say they would do something rather than ignore their peer's dangerous intentions. These relationships were mediated by students' beliefs that confiding in a teacher may have unfavorable consequences. Findings from this study support the important role schools play in creating a culture where students take responsibility for one another.


PWP1 Mediates Nutrient-Dependent Growth Control through Nucleolar Regulation of Ribosomal Gene Expression.
Ribosome biogenesis regulates animal growth and is controlled by nutrient-responsive mTOR signaling. How ribosome biogenesis is regulated during the developmental growth of animals and how nutrient-responsive signaling adjusts ribosome biogenesis in this setting have remained insufficiently understood. We uncover PWP1 as a chromatin-associated regulator of developmental growth with a conserved role in RNA polymerase I (Pol I)-mediated rRNA transcription. We further observed that PWP1 epigenetically maintains the rDNA loci in a transcription-competent state. PWP1 responds to nutrition in Drosophila larvae via mTOR signaling through gene expression and phosphorylation, which controls the nucleolar localization of dPWP1. Our data further imply that dPWP1 acts synergistically with mTOR signaling to regulate the nucleolar localization of TFIIH, a known elongation factor of Pol I. Ribosome biogenesis is often deregulated in cancer, and we demonstrate that high PWP1 levels in human head and neck squamous cell carcinoma tumors are associated with poor prognosis.


Long CTG.CAG repeat sequences markedly stimulate intramolecular recombination.
Previous studies have shown that homologous recombination is a powerful mechanism for generation of massive instabilities of the myotonic dystrophy CTG.CAG sequences. However, the frequency of recombination between the CTG.CAG tracts has not been studied. Here we performed a systematic study on the frequency of recombination between these sequences using a genetic assay based on an intramolecular plasmid system in Escherichia coli. The rate of intramolecular recombination between long CTG.CAG tracts oriented as direct repeats was extraordinarily high; recombinants were found with a frequency exceeding 12%. Recombination occurred in both RecA(+) and RecA(-) cells but was approximately 2-11 times higher in the recombination proficient strain. Long CTG.CAG tracts recombined approximately 10 times more efficiently than non-repeating control sequences of similar length. The recombination frequency was 60-fold higher for a pair of (CTG.CAG)(165) tracts compared with a pair of (CTG.CAG)(17) sequences. The CTG.CAG sequences in orientation II (CTG repeats present on a lagging strand template) recombine approximately 2-4 times more efficiently than tracts of identical length in the opposite orientation relative to the origin of replication. This orientation effect implies the involvement of DNA replication in the intramolecular recombination between CTG.CAG sequences. Thus, long CTG.CAG tracts are hot spots for genetic recombination.


The effect of dietary linoleic acid and pectin on lipoprotein and apolipoprotein A1 concentrations in rhesus monkeys.
Medium term effects (3 months) on serum lipoprotein levels of a diet with a high P/S ratio (2.2) and a low P/S ratio (0.3) were investigated in 13 normolipoproteinemic rhesus monkeys. Both diets were studied with and without added gel-forming fibre, as pectin. Addition of pectin did not have major influences on serum lipid levels. Changing from a low to a high P/S diet resulted in a significant decrease of total cholesterol (23%) and LDL cholesterol (18%) levels, but also HDL cholesterol (23%) and apolipoprotein A1 (Apo A1; 13%) concentrations fell significantly. However, after 12 weeks on the high P/S diet, HDL cholesterol concentrations rose by 24%, not significantly different from the levels during the low P/S diet. We conclude that the medium-term effects of a high P/S diet are a decrease in LDL cholesterol with only a transient effect on HDL cholesterol.


[Preventive measures against HIV infection].
The principle form of AIDS transmission are well known: sexually, blood and blood products, vertical transmission and in order to prevent HIV infections one must avoid these modes of transmission. Much has done to make more secure blood and blood products used for transfusions. It is furthermore, advised to those who must utilize intravenous drugs to employ the use of disposable syringes and more precisely, not to use the syringes of other persons. It is far more difficult to advise regarding the prevention of transmission of the virus through sexual contact, or to attempt to modify the sexual habits of certain categories of persons. Clearly, women with HIV infections should avoid pregnancy. Particular precautionary measures must be taken by those assisting patients with AIDS or HIV infections, thus avoiding infecting themselves and other patients.


Safety of JAK inhibitors: focus on cardiovascular and thromboembolic events.
To date, four Janus kinase inhibitors (JAKis) are licensed for the treatment of rheumatoid arthritis (RA) and/or psoriatic arthritis (PsA) in North America and/or Europe: tofacitinib, baricitinib, upadacitinib, and filgotinib. Most DMARDs have cardioprotective potential, yet for some of them, including JAKi, the modulatory effect on cardiovascular risk remains undetermined. Since their commercialization, the risk of venous thromboembolism (VTE) including deep vein thrombosis (DVT) and pulmonary embolism (PE) is taking on a relevant role in RA patients. In this article, we will review the effect of JAKi on CV risk and atherosclerosis process as major contributor to MACE. Furthermore, we will present the effect of JAKi on thromboembolic risk. Although the effect of JAKi on CV and thrombosis is one of the most relevant topic regarding RA patients, it is necessary to underline that these patients have an increased risk of these comorbidities due to the inflammation process, and further study are needed in order to understand the real role of JAKi in controlling or inducing these complications.


Improving image quality by optimizing beam width and helical pitch in CT.
Expanding computed tomography (CT) detector coverage broadens the beam width, but inaccurate tube current application can reduce image quality at the boundaries between body regions with different attenuation values along the z-axis. This study aims to develop and validate a new CT scanning technique with a fixed pitch to achieve higher imaging quality. A cylindrical water phantom and an anthropomorphic chest phantom with different diameters represent a human body with different attenuation values. By optimizing the beam width and helical pitch, the pitch is fixed during scanning. The mean noise of the images and the standard deviation were calculated, and the coefficient of variation (COV) was compared to evaluate the uniformity of image noise according to the beam width. At the boundaries between regions with different attenuation values, the 10 mm beam width (COV: 0.065) in the water phantom showed a 47.7% COV reduction of image noise compared with the 20 mm beam width (COV: 0.125). In addition, the 20 mm beam width (COV: 0.146) in the chest phantom showed a 29.3% COV reduction of image noise compared with the 40 mm beam width (COV: 0.206). Thus, as the beam was narrowed, the mean noise was similar, but the standard deviation was reduced. The proposed CT scanning technique with a fixed pitch, optimized beam width, and helical pitch demonstrates that image quality can be improved without increasing radiation dose at the boundary between regions with different attenuation values.


Striped phase in a quantum XY model with ring exchange.
We present quantum Monte Carlo results for a square-lattice S=1/2 XY model with a standard nearest-neighbor coupling J and a four-spin ring exchange term K. Increasing K/J, we find that the ground state spin stiffness vanishes at a critical point at which a spin gap opens and a striped bond-plaquette order emerges. At still higher K/J, this phase becomes unstable and the system develops a staggered magnetization. We discuss the quantum phase transitions between these phases.


[Hypertension, diabetes mellitus and comorbidity in primary care].
Almost every second patient seeing a primary care doctor suffers from arterial hypertension and about every fifth has diabetes mellitus. These diseases often occur at the same time. They are associated in more than 80% of the cases with other severe concomitant and subsequent diseases (heart attack, stroke, renal failure, neuropathy etc.). The magnitude especially of subsequent and concomitant diseases, the dimension of the personal suffering and the immense diagnostic and therapeutic challenges for the doctors have been massively underestimated so far. The article informs about the prevalence of the above disease, and structure of the problem. Before the background of an extremely high patient load seen by German General Physicians, the mainstay challenge is highlighted how to achieve further improvements of the quality of care on the basis of scientific guidelines alone, without a concomitant change in the system structure.


[Selective transmission of hepatitis B virus variant with S gene mutation between HBV-carrier mothers and their infants].
To investigate the role of hepatitis B virus (HBV) DNA levels in HBV-carrier mother and selective transmission of HBV variant with S gene mutation to infant in the failure of neonatal HBV immunization. Serum HBV DNA level was determined by Roche LightCycler quantities in 95 HBsAg-positive pregnant women and later in their newborns. Hepatitis B virus S gene sequences isolated from mother/child pairs were analyzed in 7 cases which did not respond to HBV vaccination and hepatitis B immunoglobulin. HBV infection was identified in 7.4 % of the infants during their first year after birth. The levels of HBV DNA in the mothers of the infected infants were higher than those of the uninfected group, and selective transmission of HBV variant from mother to infant was observed. HBV DNA levels in HBV-carrier mother and selective transmission of HBV variant from mother to infant contribute to the failure of neonatal HBV immunization.


[Emergency EEG: actual indications and results].
The indications of emergency electroencephalogram (eEEG) were defined by a French consensus conference in May 1996. We retrospectively confronted the recommendations with the actual requests for emergency EEG in our University hospital, in order to determine the contribution of the eEEG in the most frequent clinical situations encountered. Three hundred and twenty nine consecutive eEEGs over a 6-months period were retrospectively analyzed. The most frequent indications were presumption of brain death (13%), convulsive status epilepticus after treatment (12.1%), and suspicion of nonconvulsive epilepticus status (10.6%). More than one third of the requests (38.6%) were not in conformity with the recommendations of the consensus conference. The contribution of the EEG is much improved by the application of the consensual criteria. Thus, the EEG remains essential for the management of convulsive status epilepticus after treatment, to seek a subtle epilepticus status or a nonconvulsive epilepticus status. Conversely, the EEG did not prove useful in emergency after a transient loss or alteration of consciousness or a focal, non-febrile, neurological transient or permanent deficit.


Abortion foe convicted and sentenced for murders of women's health clinic workers.
On March 18, after deliberating for approximately nine hours over two days, a Dedham, Massachusetts, jury found John Salvi guilty of two counts of first-degree murder for the December 1994 shooting deaths of Shannon Lowney and Lee Ann Nichols, receptionists at two Brookline, Massachusetts, women's health care clinics. Salvi was also convicted of five counts of assault with intent to murder (see RFN IV/1). Almost immediately, Norfolk Superior Court Judge Barbara Dortch-Okara ordered Salvi to serve the maximum sentence--two consecutive life terms in prison without parole--for the murders and an additional 90-100 years for the five assaults. Under Massachusetts law, the first-degree murder convictions will be automatically appealed. In reaching their verdict, the jury of six men and six women rejected Salvi's plea of not guilty by reason of insanity. The five-week trial included testimony from more than 100 witnesses. In the past two years, two other men have been convicted for the first-degree murders of abortion providers: Michael Griffin, who was convicted of murdering Dr. David Gunn in March 1994, is now serving a life sentence; and Paul Hill, who was convicted of murdering Dr. John Britton and James Barrett in November 1995, is on Florida's death row while his sentence is pending mandatory appeal (see RFN III/19, 22).


Congenital salivary gland anlage tumor of the nasopharynx.
Nasal and upper respiratory tract obstruction in the neonatal period can result from a variety of conditions, and may be present with variable symptoms. Salivary gland anlage tumor, also referred as congenital pleomorphic adenoma, is a very rare benign congenital tumor of the nasopharynx, which may produce nasal obstruction and other associated, nonspecific symptoms. We report a case of congenital salivary gland anlage tumor causing a severe neonatal respiratory distress with pulmonary hypertension. The tumor was removed and the outcome was favourable without recurrence at five years of the follow up.


Early Responses of Sodium-Deficient Amaranthus tricolor L. Plants to Sodium Application.
Effects of sodium application on sodium-deficient Amaranthus tricolor L. cv Tricolor seedlings were studied. Thirty-day-old A. tricolor seedlings grown without sodium received either 0.5 millimolar of NaCl or KCl, and the changes in the growth rate, chlorophyll concentration, photosynthetic oxygen evolution, and dark-oxygen consumption, and some enzyme activities were compared. Following the sodium treatment, the sodium concentration in the leaves increased from the initial value of 0.4 millimolar to 2 to 3 millimolar within 24 hours, and also the relative growth rate and O(2) evolution were enhanced within 24 hours. The stimulation of O(2) evolution was greater in the upper leaves than in the lower leaves. Although total chlorophyll concentration did not increase significantly, the increase in the chlorophyll a/b ratio was apparent within 24 hours. There were not significant increases in the C(4) photosynthetic enzyme activities; however, nitrate reductase activity increased by 350% by the sodium treatment within 24 hours, and this increase is considered not to be one of the consequences of the improved photosynthesis. Results suggest that the sodium treatment promoted CO(2) and nitrate assimilation resulting in the growth enhancement, and that sodium can be involved in some other functions than C(4) photosynthesis in A. tricolor plants.


A Methylation-Based Reclassification of Bladder Cancer Based on Immune Cell Genes.
Bladder cancer is highly related to immune cell infiltration. This study aimed to develop a new classification of BC molecular subtypes based on immune-cell-associated CpG sites. The genes of 28 types of immune cells were obtained from previous studies. Then, methylation sites corresponding to immune-cell-associated genes were acquired. Differentially methylated sites (DMSs) were identified between normal samples and bladder cancer samples. Unsupervised clustering analysis of differentially methylated sites was performed to divide the sites into several subtypes. Then, the potential mechanism of different subtypes was explored. Bladder cancer patients were divided into three groups. The cluster 3 subtype had the best prognosis. Cluster 1 had the poorest prognosis. The distribution of immune cells, level of expression of checkpoints, stromal score, immune score, ESTIMATEScore, tumor purity, APC co_inhibition, APC co_stimulation, HLA, MHC class_I, Type I IFN Response, Type II IFN Response, and DNAss presented significant differences among the three subgroups. The distribution of genomic alterations was also different. The proposed classification was accurate and stable. BC patients could be divided into three subtypes based on the immune-cell-associated CpG sites. Specific biological signaling pathways, immune mechanisms, and genomic alterations were varied among the three subgroups. High-level immune infiltration was correlated with high-level methylation. The lower RNAss was associated with higher immune infiltration. The study of the intratumoral immune microenvironment may provide a new perspective for BC therapy.


ARV DRUG RESISTANCE MUTATIONS AMONG A6 SUBTYPE PLWH IN KAZAKHSTAN.
The Department of Health and Human Services and the World Health Organization (WHO) have established HIV regimens that include a variety of antiretroviral therapy (ART) medications for the treatment of HIV infected people. The Ministry of Health in Kazakhstan has authorized a clinical protocol (№97, 11.06.2020) for HIV treatment in accordance with WHO recommendations. For HIV treatment two types of ARV cocktails are recommended: preferable and alternative. The preferable is a combination of two NRTIs and INIs, whereas alternatives consist of two NRTIs' and one NNRTI. According to this protocol, ARV treatment should be administered no later than 14 days from the date of diagnosis, regardless of the clinical stage of the disease or CD4 cell count. The aim of the study was to analyze drug resistance mutations in 676 patients (52% men and 48% women) with HIV infection of subtype A6 living in different regions of Kazakhstan. ARV therapy was received by 18% of patients for less than 3 years; 48% - for 3-5 years; 24% for 6-8 years. The preferable ARV therapy was received by 49% of patients; 42.4% of patients received alternative therapy; 8.6% of patients took other types of cocktails. It was concluded that the main reason for the development of mutations of resistance to ARV drugs is insufficient adherence to the ARV therapy regimen. This may be due to difficulties in accessing medical care, stigmatization of HIV and a shortage of medicines due to exorbitant costs or other factors. The development of mutations of resistance to ARV drugs makes it difficult to maintain viral load suppression, creating a problem for the health of patients and at the same time increasing the risk of further HIV transmission.


Crossed cerebellar hyperperfusion in symptomatic epilepsy--two case reports.
A 74-year-old female with cerebral infarction and a 78-year-old female with cerebral glioblastoma suffered complex partial seizure. Ictal perfusion single photon emission computed tomography in these patients showed the interesting phenomenon of 'crossed cerebellar hyperperfusion,' a reversed crossed cerebellar diaschisis. The mechanism is probably spread of electrical seizure through efferent projections, and may be related to the cerebellar atrophy seen in patients with long-standing partial epilepsy.


MR of the spinal cord in a patient with herpes zoster.
Increased signal intensity on initial magnetic resonance images of the spinal cord in a patient with herpes zoster demonstrated that this virus caused inflammation of the cervical spinal cord. This pathology corresponded well with neurologic deficits seen clinically, but the extent of the neurologic deficits ultimately could not be determined by magnetic resonance of the spinal cord alone because the nerve roots were also affected.


Viral loads in clinical specimens and SARS manifestations.
A retrospective viral load study was performed on clinical specimens from 154 patients with laboratory-confirmed severe acute respiratory syndrome (SARS); the specimens were prospectively collected during patients' illness. Viral load in nasopharyngeal aspirates (n = 142) from day 10 to day 15 after onset of symptoms was associated with oxygen desaturation, mechanical ventilation, diarrhea, hepatic dysfunction, and death. Serum viral load (n = 53) was associated with oxygen desaturation, mechanical ventilation, and death. Stool viral load (n = 94) was associated with diarrhea, and urine viral load (n = 111) was associated with abnormal urinalysis results. Viral replications at different sites are important in the pathogenesis of clinical and laboratory abnormalities of SARS.


Bacterial delivery of RNAi effectors: transkingdom RNAi.
RNA interference (RNAi) represents a high effective mechanism for specific inhibition of mRNA expression. Besides its potential as a powerful laboratory tool, the RNAi pathway appears to be promising for therapeutic utilization. For development of RNA interference (RNAi)-based therapies, delivery of RNAi-mediating agents to target cells is one of the major obstacles. A novel strategy to overcome this hurdle is transkingdom RNAi (tkRNAi). This technology uses non-pathogenic bacteria, e.g. Escherichia coli, to produce and deliver therapeutic short hairpin RNA (shRNA) into target cells to induce RNAi. A first-generation tkRNAi-mediating vector, TRIP, contains the bacteriophage T7 promoter for expression regulation of a therapeutic shRNA of interest. Furthermore, TRIP has the Inv locus from Yersinia pseudotuberculosis that encodes invasin, which permits natural noninvasive bacteria to enter beta1-integrin-positive mammalian cells and the HlyA gene from Listeria monocytogenes, which produces listeriolysin O. This enzyme allows the therapeutic shRNA to escape from entry vesicles within the cytoplasm of the target cell. TRIP constructs are introduced into a competent non-pathogenic Escherichia coli strain, which encodes T7 RNA polymerase necessary for the T7 promoter-driven synthesis of shRNAs. A well-characterized cancer-associated target molecule for different RNAi strategies is ABCB1 (MDR1/P-glycoprotein, MDR1/P-gp). This ABC-transporter acts as a drug extrusion pump and mediates the "classical" ABCB1-mediated multidrug resistance (MDR) phenotype of human cancer cells which is characterized by a specific cross resistance pattern. Different ABCB1-expressing MDR cancer cells were treated with anti-ABCB1 shRNA expression vector bearing E. coli. This procedure resulted in activation of the RNAi pathways within the cancer cells and a considerable down regulation of the ABCB1 encoding mRNA as well as the corresponding drug extrusion pump. Accordingly, drug accumulation was enhanced in the pristine drug-resistant cancer cells and the MDR phenotype was reversed. By means of this model the data provide the proof-of-concept that tkRNAi is suitable for modulation of cancer-associated factors, e.g. ABCB1, in human cancer cells.


Profiling the safety and tolerability of bisphosphonates.
Because patients with metastatic bone disease suffer a significant burden from their illness and from anticancer treatments, it is therefore important to minimize the side effects of bisphosphonates. The intravenous bisphosphonates, zoledronic acid and pamidronate, have tolerability issues that include a flu-like syndrome, injection-site reactions, and occasional renal toxicity. Because of the potentially severe nature of the renal toxicity, renal monitoring is required before each dose, with drug withdrawal if the patients' renal function deteriorates. Oral clodronate often causes gastrointestinal disturbances, particularly diarrhea; compliance is often poor because of the large tablet size and multiple daily dosing. Long-term data have shown that the bisphosphonate ibandronate is well tolerated either intravenously or orally, with a renal safety profile similar to placebo and no evidence of cumulative renal damage. Studies investigating the effects of 15-minute infusions and intensive dosing indicate that intravenous ibandronate given rapidly or at high doses is also well tolerated with no renal safety concerns. Taken together, these results suggest that the favorable safety profile of ibandronate provides an important alternative to existing bisphosphonate options for metastatic bone disease. Using ibandronate could improve patient acceptability and simplify management, with reductions in the need for safety monitoring and management of adverse events.


[Spleen-preserving distal pancreatectomy in treatment of solid-pseudopapillary neoplasm].
To investigate the feasibility and safety of spleen-preserving distal pancreatectomy with conservation of the splenic arteries and veins in treatment of benign neoplasms of distal pancreas. Four patients with solid-pseudopapillary neoplasm, 1 males and 3 females, aged 30 (17 - 37), underwent laparotomy. The distal pancreas with tumor was cut. The perforating branches between the splenic artery and vein and the distal pancreas were isolated, ligated, and cut. The splenic artery and vein and the short and left gastric vessels were all preserved. The spleen remained intact. The mean operation time was 208 minutes +/- 52 minutes, and the mean blood loss was 475 ml +/- 96 ml. Blood transfusion was not necessary. The mean post-operative hospital stay was 18 days +/- 13 days. No post-operative complications were found except for pancreatic leakage occurring in one patient that was cured by conservative treatment. No metastasis and recurrence was found. Safe and feasible, spleen-preserving distal pancreatectomy with conservation of the splenic arteries and veins is one of the best choices for treatment of benign and borderline tumors of pancreas.


Improved experimental procedures for achieving efficient germ line transmission of nonobese diabetic (NOD)-derived embryonic stem cells.
The manipulation of a specific gene in NOD mice, the best animal model for insulin-dependent diabetes mellitus (IDDM), must allow for the precise characterization of the functional involvement of its encoded molecule in the pathogenesis of the disease. Although this has been attempted by the cross-breeding of NOD mice with many gene knockout mice originally created on the 129 or C57BL/6 strain background, the interpretation of the resulting phenotype(s) has often been confusing due to the possibility of a known or unknown disease susceptibility locus (e.g., Idd locus) cosegregating with the targeted gene from the diabetes-resistant strain. Therefore, it is important to generate mutant mice on a pure NOD background by using NOD-derived embryonic stem (ES) cells. By using the NOD ES cell line established by Nagafuchi and colleagues in 1999 (FEBS Lett., 455, 101-104), the authors reexamined various conditions in the context of cell culture, DNA transfection, and blastocyst injection, and achieved a markedly improved transmission efficiency of these NOD ES cells into the mouse germ line. These modifications will enable gene targeting on a "pure" NOD background with high efficiency, and contribute to clarifying the physiological roles of a variety of genes in the disease course of IDDM.


[Extended total gastrectomy: indications in the 3rd millennium].
Total extended gastrectomy (TEG) is indicated in the treatment of gastric cancer for necessity or to achieve an oncologic radicality. By this surgical treatment the stomach and other organs or a part of them involved by primitive tumor are removed. The authors report a study about 15 patients, out of 116 cases of gastric cancer, operated by TEG between 1990-1998. The middle-age of this patients was 63 years (range 45-76) and their general conditions were good in 9 cases and not-good in 6. The postoperative total parenteral nutrition (TPN) was carried out in all the patients, while preoperatively only in the most compromised patients. The surgical treatments were: 2 TG (total gastrectomy)+splenecomy; 3 TG+splenectomy+pancreatic resection; 4 TG+splenectomy+pancreatic resection+distal esophageal resection; 1 TG+distal esophageal resection; 2 TG+atypic hepatic resection; 1 TG+ atypic hepatic resection+duodenum resection; 2 TG+large intestine resection. While 10 patients were operated on to obtain radicality, 5 patients had a palliative treatment. There was not perioperative mortality, but we have observed: one dehiscence of the duodenal stump and one pancreatic fistula treated with conservative therapy; one left subfrenic abscess treated with surgical therapy. The survival has been higher in the patients treated with radicality. On the basis of these cases, the authors consider: 1) the possibility to obtain radicality by TEG; 2) the gastric localizations more often associated to extravisceral neoplastic localization; 3) the role of extensive lymph node resection (III and IV level) to obtain oncological radicality or neoplastic reduction. CONCLUSIONS. On the basis of their personal experience and related literature, the authors conclude that TEG is indicated to: 1) obtain a better lymphadenectomy; 2) obtain an oncologic radicality; 3) reduce the neoplastic mass in order to facilitate adjuvant therapy; 4) avoid or treat neoplastic complications; 5) improve the quality of life.


Total syntheses of (+)-chloropuupehenone and (+)-chloropuupehenol and their analogues and evaluation of their bioactivities.
Tetracyclic pyrans (+)-chloropuupehenone (1) and (+)-chloropuupehenol (5) and its C8-R-isomer (+)-3 were synthesized via a one-pot condensation of 1-chloro-2-lithio-3,5,6-tris(tert-butyldimethylsilyloxy)benzene (8) with (4aS,8aS)-3,4,4a,5,6,7,8,8a-octahydro-2,5,5,8a-tetramethylnaphthalene-1-carboxaldehyde (7). The major condensation product, (4aS,6aR,12bS)-2H-9,10-bis(tert-butyldimethylsilyloxy)-11-chloro-1,3,4,4a,5,6,6a,12b-octahydro-4,4,6a,12b-tetramethyl-benzo[a]xanthene (4), after desilylation provided tetracyclic pyran (+)-(4aS,6aR,12bS)-2H-11-chloro-1,3,4,4a,5,6,6a,12b-octahydro-4,4,6a,12b-tetramethyl-benzo[a]xanthene-9,10-diol (3). At a dosage of 42 mg/rat over 8 h, pyran diol 3 inhibited the intestinal absorption of cholesterol by 71% in rats. Tetracyclic pyran 4 was also converted to o-quinone 28, which inhibited cholesteryl ester transfer protein (CETP) activity and L1210 leukemic cell viability with IC(50) values of 31 and 2.4 microM, respectively. Diol (+)-5 inhibited CETP activity with an IC(50) value of 16 microM. The minor condensation product, (4aS,6aS,12bS)-2H-9,10-bis(tert-butyldimethylsilyloxy)-11-chloro-1,3,4,4a,5,6,6a,12b-octahydro-4,4,6a,12b-tetramethyl-benzo[a]xanthene (6), was transformed into (+)-5 and (+)-1. A stepwise stereoselective synthesis of (+)-1 was also developed utilizing an oxyselenylation ring-closure reaction. The synthetic sequence also produced four biologically active naturally occurring drimanic sesquiterpenes, (+)-drimane-8alpha,11-diol (34), (-)-drimenol (38), (+)-albicanol (39), and (-)-albicanal (31) as intermediates.


The NAC domain mediates functional specificity of CUP-SHAPED COTYLEDON proteins.
In higher plants, although several genes involved in shoot apical meristem (SAM) formation and organ separation have been isolated, the molecular mechanisms by which they function are largely unknown. CUP-SHAPED COTYLEDON (CUC) 1 and CUC2 are examples of two such genes that encode the NAC domain proteins. This study investigated the molecular basis for their activities. Nuclear localization assays indicated that green fluorescent protein (GFP)-CUC proteins accumulate in the nucleus. Yeast one-hybrid and transient expression assays demonstrated that the C-terminal domain (CTD) of the CUC has transactivation activity. Domain-swapping experiments revealed that the functional specificity of the CUC for promoting adventitious shoot formation resides in the highly conserved NAC domain, not in the CTD in which motifs specific to the CUC subfamily are located. Taken together, these observations suggest that CUC proteins transactivate the target genes involved in SAM formation and organ separation through a specific interaction between the NAC domain and the promoter region of the target genes.


Targeting vascular endothelial growth factor receptor in thyroid cancer: the intracellular and extracellular implications.
Our understanding of the molecular pathophysiology of differentiated thyroid cancer (DTC) has developed considerably over the last 10 years. Aberrant signaling through B-Raf and Akt has been implicated in the tumorigenesis of DTC. Moreover, these highly vascular tumors have proven to be sensitive to the inhibition of vascular endothelial growth factor receptor (VEGFR-2). It is likely that the multikinase inhibitors, sorafenib, sunitinib, axitinib, and motesanib, whose targets include VEGFR-2, exert their effects primarily through inhibition of endothelial cells. However, as VEGFR-2 is expressed on DTC cells, these compounds may have direct antitumor action. This review will discuss the key signaling pathways involved in thyroid cancer and their implications for targeted therapy.


Drug-Drug Interaction Studies of Methadone and Antiviral Drugs: Lessons Learned.
Different views appear in the literature on the extent of specific cytochrome P450 (CYP) involvement in methadone metabolism. The aim of this work is to leverage knowledge from drug-drug interaction (DDI) studies in new drug applications between methadone and antiviral medications to better understand methadone disposition and to inform design of future DDI studies with methadone. A database of DDI studies between all FDA-approved human immunodeficiency virus and hepatitis C virus medications and methadone was constructed. The database contains data from 29 DDI studies. Sixteen of the 29 studies had statistically significant changes in methadone area under the concentration-time curve. Methadone exposure was either decreased or unchanged when it was coadministered with weak to strong CYP3A inhibitors or a moderate CYP3A4 inducer. Methadone exposure was reduced when it was coadministered with CYP2B6 inducers. The role of other enzymes (CYP2C9, CYP2C19, and CYP2D6) cannot be fully elucidated from these studies. In conclusion, CYP2B6 plays a prominent role in methadone metabolism, although methadone exposure is not sensitive to CYP3A perturbation. In designing methadone DDI studies, (1) measuring R- and S-methadone is more informative than measuring total methadone, and (2) CYP2B6 genotyping of subjects enrolled in methadone DDI studies should be considered. Finally, there is a need for the development of predictive models to determine the influence of medications on methadone disposition.


Insulin-like growth factor-II: its role in metabolic and endocrine disease.
Insulin-like growth factor-II (IGF-II) is a widely expressed 7·5 kDa mitogenic peptide hormone. Although it is abundant in serum, understanding of its physiological role is limited compared with that of IGF-I. IGF-II regulates foetal development and differentiation, but its role in adults is less well understood. Evidence suggests roles in a number of tissues including skeletal muscle, adipose tissue, bone and ovary. Altered IGF-II expression has been observed in metabolic conditions, notably obesity, diabetes and the polycystic ovary syndrome. This article summarizes what is known about the actions of IGF-II and its dysregulation in metabolic and endocrine diseases. The possible causes and consequences of dysregulation are discussed along with the implications for diagnostic tests and future research.


Epigenetic factors up-regulate expression of myelin proteins in the dysmyelinating jimpy mutant mouse.
Proteolipid protein (PLP) is a major structural component of central nervous system (CNS) myelin. Evidence exists that PLP or the related splice variant DM-20 protein may also play a role in early development of oligodendrocytes (OLs), the cells that form CNS myelin. There are several naturally occurring mutations of the PLP gene that have been used to study the roles of PLP both in myelination and in OL differentiation. The PLP mutation in the jimpy (jp) mouse has been extensively characterized. These mutants produce no detectable PLP and exhibit an almost total lack of CNS myelin. Additionally, most OLs in affected animals die prematurely, before producing myelin sheaths. We have studied cultures of jp CNS in order to understand whether OL survival and myelin formation require production of normal PLP. When grown in primary cultures, jp OLs mimic the relatively undifferentiated phenotype of jp OLs in vivo. They produce little myelin basic protein (MBP), never immunostain for PLP, and rarely elaborate myelin-like membranes. We report here that jp OLs grown in medium conditioned by normal astrocytes synthesize MBP and incorporate it into membrane expansions. Some jp OLs grown in this way stain with PLP antibodies, including an antibody to a peptide sequence specific for the mutant jp PLP. This study shows that: (1) an absence of PLP does not necessarily lead to dysmyelination or OL death; (2) OLs are capable of translating at least a portion of the predicted jp PLP; (3) the abnormal PLP made in the cultured jp cells is not toxic to OLs. These results also highlight the importance of environmental factors in controlling OL phenotype.


International Society for Cell and Gene Therapy of Cancer 2009 Annual Meeting held in Cork, Ireland.
The International Society for Cell and Gene Therapy (ISCGT) of Cancer annual meeting was held from September 2 through September 4, 2009, in Cork, Ireland ( www.iscgt2009.com ). The conference was held in conjunction with the Irish Society for Gene and Cell Therapy third annual meeting, and brought together scientists and clinicians from around the world in a country developing its knowledge economy. Next year's ISCGT meeting will be held in Doha, the capital of Qatar ( www.iscgt.net ), from September 27 through September 29, 2010.


Genomic resources for wild populations of the house mouse, Mus musculus and its close relative Mus spretus.
Wild populations of the house mouse (Mus musculus) represent the raw genetic material for the classical inbred strains in biomedical research and are a major model system for evolutionary biology. We provide whole genome sequencing data of individuals representing natural populations of M. m. domesticus (24 individuals from 3 populations), M. m. helgolandicus (3 individuals), M. m. musculus (22 individuals from 3 populations) and M. spretus (8 individuals from one population). We use a single pipeline to map and call variants for these individuals and also include 10 additional individuals of M. m. castaneus for which genomic data are publically available. In addition, RNAseq data were obtained from 10 tissues of up to eight adult individuals from each of the three M. m. domesticus populations for which genomic data were collected. Data and analyses are presented via tracks viewable in the UCSC or IGV genome browsers. We also provide information on available outbred stocks and instructions on how to keep them in the laboratory.


Genomic Analysis of a New Serovar of Leptospira weilii Serogroup Manhao.
Leptospirosis, caused by pathogenic Leptospira spp., is recognized as an important emerging zoonotic disease throughout the world. In this study, multiple approaches were used to characterize the recently discovered serovar Heyan strain L231. This strain can infect guinea pigs and belonged to the pathogenic species L. weilii. Genome sequencing analysis revealed the draft genome of 4.2 M bp with a G+C content of 40.67% for strain L231, and a total of 4,794 ORFs were identified. The strain L231 genome was found to have a larger LPS biosynthesis locus than that of strains L. interrogans serovar Lai and L. borgpetersenii serovar Hardjobovis. Phylogenomic reconstructions showed that the evolutionary position of L. weilii serovar Heyan was different from that of other serovars from serogroup Manhao. These findings may lead us to a better understanding of Leptospira pathogenesis and evolution.


Antiretroviral Therapy for HIV Infection Induced-Neuropathic Pain, A Narrative Review.
Human immunodeficiency virus (HIV) causes acquired immunodeficiency syndrome (AIDS). AIDS is a human disease in which there is a systematic failure of the immune system, thereby leading to severe opportunistic infections. HIV is treated with antiretroviral therapy (ART), which helps in preventing the virus from replicating in the body. ART also enables the immune system to repair itself and restrict further injury. Nevertheless, the long-term use of ART leads to multiple neurological complications (e.g.neuropathic pain). Neuropathic pain (NP) arises if the nervous system is damaged or not working correctly. NP is is characterized by pain in both hands and feet and can hinder the quality of life (QOL) as it is always linked to impaired cognition, anxiety, depression, loss of function, among others. To provide an overview of the clinical issues regarding the toxicity implications arising from the long-tern use of antiretroviral therapy (ART). In this paper, we ascertained and summarized studies on manifestations of ART-induced neuropathic pain (ART-NP)-using the relevant and up-to-date facts from Google Scholar, and PubMed, among others. While ART assists in suppressing HIV and may help to lessen neurocognitive impairments, long-term use incurs toxicity to the central nervous system (CNS). Consequently, future studies must explore the effects of ART on adaptations in distinct regions of the CNS as well as the pathophysiological feature of ART-induced NP.


The General Approach and Management of the Patient Who Discloses a Sexual Assault.
The effects a sexual assault on a survivor can be profound and multifaceted. Some of the aftermath may include bodily and/or anal/genital injury, sexually transmitted infection (STI) or disease (STD), post-traumatic stress disorder (PTSD), depression, suicidal ideation, and pregnancy. There is no typical experience or survivor response. So, if a survivor presents to the Emergency Departments for care, it is imperative for healing that the response is stabilizing, coordinated, and compassionate. Immediate needs of a survivor of sexual assault include: medical and/or psychiatric evaluation and stabilization, activation of community advocacy to the bedside, mandated reporting as directed by state statutes, offering and conducting (if desired by the survivor) the Sexual Assault Forensic Exam (SAFE), collaboration with law enforcement (if desired by the survivor), prophylactic medications for STI and STD, pregnancy risk evaluation and care and safe discharge planning.


Synthesis of 3-hydroxyoxindoles by Pd-catalysed intramolecular nucleophilic addition of aryl halides to alpha-ketoamides.
Pd/PtBu(3)-catalysed intramolecular nucleophilic addition of aryl halides to alpha-ketoamides in the presence of nBuOH and base has been realized with high yields, providing a new, direct, and efficient synthetic strategy to obtain 3-hydroxyoxindoles.


Effect of different beta-blocking drugs and adrenaline on the conversion of thyroxine to triiodothyronine in isolated rat hepatocytes.
The in vitro effect of various selective and non-selective beta-blocking drugs and adrenaline on the conversion of thyroxine (T4) to triiodothyronine (T3) was studied in suspensions of isolated rat hepatocytes after 90 min of incubation. Compared with the untreated controls propranolol caused a dose-related inhibition of the T4 to T3 conversion in conc of 100, 200 and 400 microM. The other beta-blocking drugs studied, timolol, oxprenolol, atenolol and metoprolol, were without any effect on this in vitro conversion. Propranolol did not interfere with the cellular association of T4 or the degradation of T4 and T3. Adrenaline 200 microM caused a small decrease of T3 in the medium and a corresponding increase in the intracellular content of T3. The inhibitory effect of propranolol 200 microM was not antagonized by equimolar concentrations of adrenaline. Our study suggests that the inhibitory effect of propranolol on the conversion of T4 to T3 in hepatocytes is caused by a direct chemical effect of the drug unrelated to its beta-blocking and membrane stabilizing properties.


Design Analysis of Adhesively Bonded Structures.
The existing analytical solutions for the peeling and shearing stresses in polymeric adhesively bonded structures are either too inaccurate or too complex for adoption by practicing engineers. This manuscript presents a closed-form solution that is reasonably accurate yet simple and concise enough to be adopted by practicing engineers for design analysis and exploration. Analysis of these concise solutions have yielded insightful design guidelines: (i) the magnitude of peeling stress is generally higher than that of shearing stress; (ii) the peeling stress in a balanced structure may be reduced most effectively by reducing the elastic modulus of the adherends or by increasing the adhesive-to-adherend thickness ratio and less effectively by reducing the elastic modulus of the adhesive; and (iii) the peeling stress in an unbalanced structure may be reduced by increasing the in-plane compliance of the structure, which may be implemented most effectively by reducing the thicknesses of the adherends and less effectively by reducing the elastic modulus of the adherends.


Effect of eutectic mixture of local anesthetics on pain perception during scaling by ultrasonic or hand instruments: a masked randomized controlled trial.
The aims of this study are to evaluate the efficacy of a eutectic mixture of local anesthetic (EMLA) cream on pain perception during scaling and to compare the intensities of pain provoked by hand and ultrasonic instruments. Forty subjects with chronic gingivitis or periodontitis were enrolled in the study. In this randomized, split-mouth, controlled, masked clinical trial, each of the four quadrants in subjects was randomly assigned to receive one of the following protocols: scaling with an ultrasonic scaler with or without the application of the EMLA cream or scaling with a Gracey curet with or without the application of the EMLA cream. Pain levels after each quadrant of scaling were assessed with a visual analog scale (VAS; 0 to 100 mm) and verbal rating score (VRS; 0 to 4). All subjects were recalled to detect any complications after 24 hours. The mean VAS and VRS when EMLA cream was applied (18.39 ± 14.47 mm and 0.95 ± 0.69) were significantly lower (P <0.001 for VAS and VRS) compared to when EMLA cream was not used (26.54 ± 16.46 mm and 1.30 ± 0.75). The mean VAS and VRS of the ultrasonic scaler group (20.43 ± 16.40 mm and 0.98 ± 0.76) were significantly lower (P = 0.024 for VAS; P = 0.003 for VRS) than those of the Gracey curet group (24.50 ± 15.17 mm and 1.28 ± 0.69). None of the subjects showed adverse effects after EMLA-cream application. Although most patients experienced limited pain during scaling, a significant reduction of pain is achieved by using EMLA cream and ultrasonic scaler.


SEM study on the dorsal lingual surface of the large flying fox, Pteropus vampyrus.
The dorsal lingual surface of the large flying foxes were examined by scanning electron microscopy. Filiform, fungiform and vallate papillae were observed. The filiform papillae were distributed over the entire dorsal surface of the tongue. The filiform papillae notably differed in morphology by their location on the tongue and could be classified into 6 types: 1) scale-like, 2) giant trifid, 3) small crown-like, 4) large crown-like, 5) long conical and 6) short conical papillae. The fungiform papillae were present rounded bodies on the anterior 2/3 of the tongue. The large flying foxes showed the triangular arrangement of the three vallate papillae, with the apex of the triangle directed posteriorly. These findings indicate that the tongue of the large flying bat is similar to that of the lesser dog-faced fruit bat.


Fenretinide, Tocilizumab, and Reparixin Provide Multifaceted Disruption of Oral Squamous Cell Carcinoma Stem Cell Properties: Implications for Tertiary Chemoprevention.
Locoregional recurrence of oral squamous cell carcinoma (OSCC) dramatically reduces patient survival. Further, as many OSCC recurrences are inoperable, radiotherapy and chemotherapy with or without biological adjuncts are the remaining treatment options. Although the tumors may initially respond, radiotherapy- and chemotherapy-resistant cancer stem cells (CSC) can readily repopulate OSCC tumors. Currently, following the initial OSCC treatment, patients are closely monitored until a recurrence or a second primary is detected. Identification of agents with complementary mechanisms to suppress CSC tumorigenic functions could change this passive approach. The goals of this study were twofold: (1) develop and validate CSC-enriched (CSCE) OSCC cell lines and (2) identify chemopreventive agents that obstruct multiple CSCE protumorigenic pathways. CSCE cultures, which were created by paclitaxel treatment followed by three tumorsphere passes, demonstrated CSC characteristics, including increased expression of stem cell and inflammatory genes, increased aldehyde dehydrogenase (ALDH) activity, and enhanced in vitro/in vivo proliferation and invasion. Three chemopreventives, fenretinide, tocilizumab, and reparixin, were selected due to their distinct and complementary CSC-disruptive mechanisms. The CSCE selection process modulated the cells' intermediate filaments resulting in an epithelial-predominant (enhanced cytokeratin, proliferation, IL6 release) line and a mesenchymal-predominant (upregulated vimentin, invasive, IL8 release) line. Our results confirm that 4HPR binds with appreciably higher affinity than Wnt at the Frizzled binding site and significantly inhibits CSC-enabling Wnt-β-catenin downstream signaling. Notably, combination fenretinide-tocilizumab-reparixin treatment significantly suppressed IL6 and IL8 release, stem cell gene expression, and invasion in these diverse CSCE populations. These promising multiagent in vitro data provide the basis for our upcoming in vivo CSCE tertiary chemoprevention studies.


[Prenatal diagnosis of lethal junctional epidermolysis bullosa by fetal skin biopsy].
A 22-year-old woman, whose first infant had died of lethal junctional epidermolysis bullosa (JEB), requested prenatal diagnosis for her third pregnancy. At 20 weeks gestation, fetal biopsy was performed under direct vision by fetoscopy. A semithin section of epon-embedded skin showed dermo-epidermal separation at the light microscopic level. Electron microscopy revealed the site of separation to be within the lamina lucida of the epidermal basement membrane (EBM). Indirect immunofluorescence on a 5 microns cryostat specimen of skin showed a complete absence of GB3 monoclonal antibody immunostaining at the EBM compared with a control 18 week old normal fetal skin sample. The diagnosis was therefore made that the fetus was affected with lethal JEB and a prostaglandin termination performed. The diagnosis was confirmed by further studies on the aborted fetus. 54 cases of prenatal diagnosis of various types of epidermolysis bullosa performed at Institute of Dermatology over the last 10 years are briefly reviewed. Several social and practical problems to launch prenatal diagnosis in Japan are also discussed.


Impacts of single-walled carbon nanotubes on microbial community structure in activated sludge.
Single-walled carbon nanotubes (SWNTs) are likely to become increasingly widespread and yet their environmental impact is not well understood. The purpose of the current study was to evaluate the impact of SWNTs on microbial communities in a 'sentinel' environmental system, activated sludge batch-scale reactors. Triplicate batch reactors were exposed to SWNTs and compared to control reactors exposed to impurities associated with SWNTs. Automated ribosomal intergenic spacer analysis (ARISA) was used to assess bacterial community structure in each reactor. SWNT exposure was found to impact microbial community structure, while SWNT-associated impurities had no effect, compared to controls. 16S rRNA gene sequence analysis indicated that dominant phylotypes detected by ARISA included members of the families Sphingomonadaceae and Cytophagacaceae and the genus Zoogloea. ARISA results indicated an adverse impact of SWNTs on the sphingomonad relative to other community members. Changes in community structure also occurred in both SWNT-exposed and control reactors over the experimental time period and with the date on which activated sludge was obtained from a wastewater treatment facility. These results indicate that SWNTs differentially impact members of the activated sludge reactor bacterial community. The finding that community structure was affected by SWNTs indicates that this emerging contaminant differentially impacted members of the activated sludge bacterial community and raises the concern that SWNTs may also affect the services it provides.


Cardioprotective Effects of Genistin in Rat Myocardial Ischemia-Reperfusion Injury Studies by Regulation of P2X7/NF-κB Pathway.
The present study aimed to assess the effects and mechanisms of genistin in the rat model of myocardial ischemia reperfusion injury. The rat hearts were exposed to the left anterior descending coronary artery (LAD) ligation for 30 min followed by 1 h of reperfusion. In the rat of myocardial ischemia/reperfusion (MI/R), it was found that genistin pretreatment reduced myocardial infarct size, improved the heart rate, and decreased creatine kinase (CK) and lactate dehydrogenase (LDH) levels in coronary flow. This pretreatment also increased catalase (CAT), superoxide dismutase (SOD) activities but decreased glutathione (GSH), malondialdehyde (MDA) levels. Furthermore, we determined that genistin can ameliorate the impaired mitochondrial morphology and oxidation system; interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10), and tumor necrosis factor-α (TNF-α) levels were also recovered. Besides, related-proteins of nuclear factor kappa-B (NF-κB) signal pathway activated by P2X7 were investigated to determine the molecular mechanism of genistin and their expressions were measured by western blot. These results presented here demonstrated that genistin enhanced the protective effect on the rats with myocardial ischemia reperfusion injury. Therefore, the cardioprotective effects of genistin may rely on its antioxidant and anti-inflammatory activities via suppression of P2X7/NF-κB pathways.


Synergistic Effects of 3-Nitrooxypropanol with Fumarate in the Regulation of Propionate Formation and Methanogenesis in Dairy Cows In Vitro.
3-Nitrooxypropanol (3-NOP) is effective at reducing ruminal methane emissions in ruminants. But it also causes a drastic increase in hydrogen accumulation, resulting in feed energy waste. Fumarate is a key precursor for propionate formation and plays an important role in rumen hydrogen metabolism. Therefore, this study examined the effects of 3-NOP combined with fumarate on volatile fatty acids, methanogenesis, and microbial community structures in dairy cows in vitro. The in vitro culture experiment was performed using a 2-by-2 factorial design, two 3-NOP levels (0 or 2 mg/g dry matter [DM]) and two fumarate levels (0 or 100 mg/g DM), including 3 runs with 4 treatments, 4 replicates, and 4 blanks containing only the inoculum. Rumen fluid was collected from three lactating Holstein cows with permanent ruminal fistulas. The combination of 3-NOP and fumarate reduced methane emissions by 11.48% without affecting dry matter degradability. The propionate concentration increased and the acetate/propionate ratio decreased significantly. In terms of bacteria, the combination of 3-NOP and fumarate reduced the abundances of Ruminococcus and Lachnospiraceae_NK3A20_group and increased the abundances of Prevotella and Succiniclasticum. For archaea, the combination of 3-NOP and fumarate significantly increased the abundances of Methanobrevibacter_sp._AbM4, while the abundance of operational taxonomic unit 581 (OTU581) (belonging to an uncultured_rumen_methanogen_g__Methanobrevibacter strain) was significantly decreased. These results indicated that the combination of 3-NOP and fumarate could alleviate the accumulation of hydrogen and enhance the inhibition of methanogenesis compared with 3-NOP only in dairy cows. IMPORTANCE The global problem of climate change and the greenhouse effect has become increasingly severe, and the abatement of greenhouse gases has received great attention from the international community. Methane produced by ruminants during digestion not only aggravates the greenhouse effect but also causes a waste of feed energy. As a methane inhibitor, 3-nitrooxypropanol can effectively reduce methane emissions from ruminants. However, when it inhibits methane emissions, the emission of hydrogen increases sharply, resulting in the waste of feed resources. Fumarate is a propionic acid precursor that can promote the metabolism of hydrogen to propionic acid in animals. Therefore, we studied the effects of the combined addition of 3-nitrooxypropanol and fumarate on methanogenesis, rumen fermentation, and rumen flora. It is of great significance to inhibit methane emission from ruminants and slow down the greenhouse effect.


Determination of Polychlorinated Biphenyls in Solid Samples by Isotope Dilution Mass Spectrometry Using ³⁷Cl-Labeled Analogues.
This work describes the first application of (37)Cl-labeled compounds to isotope dilution mass spectrometry (IDMS). The synthesis of 12 (37)Cl-labeled polychlorinated biphenyls (PCBs) was carried out by the chlorination of biphenyl with isotopically enriched chlorine gas, generated by the direct oxidation of Na(37)Cl with potassium peroxymonosulfate. After an exhaustive purification due to the presence of other congeners, the concentration and the isotopic enrichment of all (37)Cl-labeled PCBs in the mixture was determined. The proposed procedure allows the simultaneous quantification of every isotope diluted PCB congener in a single gas chromatography-tandem mass spectrometry (GC-MS/MS) injection without resorting to a methodological calibration graph. The results obtained here demonstrate that the use of (37)Cl-labeled analogues provides results in agreement with the certified values of three different Certified Reference Materials (marine sediment SRM 1944, fish tissue 1947, and loamy soil CRM 962-50) and analytical figures of merit comparable to those obtained using regular IDMS procedures based on the use of commercially available (13)C-labeled analogues.


Practical Imaging Strategies for Intussusception in Children.
OBJECTIVE. Intussusception is the most common cause of intestinal obstruction in young children. Radiology has a key role in its diagnosis and treatment. This systematic review summarizes the currently available evidence for best practices in radiologic management of pediatric intussusception. CONCLUSION. High diagnostic accuracy and lack of ionizing radiation make ultrasound (US) the preferred imaging modality for diagnosing intussusception. For intussusception reduction, fluoroscopy-guided pneumatic enema and US-guided hydrostatic enema are equally dependable and safe techniques. The areas that warrant further research in this field include the efficacy and safety of the US-guided pneumatic enema, potential benefits of sedation and general anesthesia for the reduction procedure, and the optimal management of intussusceptions potentially involving pathologic lead points.


Seven-color time-resolved fluorescence hybridization analysis of human papilloma virus types.
Identification of human papilloma virus (HPV) types is important in order to determine the risk of cervical carcinoma in women. This requires a technique to probe individual samples for multiple virus specificities. Here we describe simultaneous multicolor analysis of amplification products for any of seven amplified HPV types 16, 18, 31, 33, 35, 39, and 45, associated with cancer of the cervix. A seminested polymerase chain reaction was performed in a single tube using a biotinylated inner primer. Sets of amplification products, immobilized on a 96-pronged manifold solid support, were rendered single stranded and probed with a mix of seven type-specific, differentially labeled oligonucleotides. These probes contained 10 or 20 lanthanide chelates at the 5' ends with seven distinct combinations of europium, terbium, and samarium ions. The seven viral strains were correctly identified by time-resolved fluorescence measurement of the specifically hybridized probes. Using this assay format, simultaneous detection of any of seven or even more target variants is possible.