Stimulus generalization of conditioned eyelid responses produced without cerebellar cortex: implications for plasticity in the cerebellar nuclei.
In Pavlovian eyelid conditioning and adaptation of the vestibulo-ocular reflex, cerebellar cortex lesions fail to completely abolish previously acquired learning, indicating an additional site of plasticity in the deep cerebellar or vestibular nucleus. Three forms of plasticity are known to occur in the deep cerebellar nuclei: formation of new synapses, plasticity at existing synapses, and changes in intrinsic excitability. Only a cell-wide increase in excitability predicts that learning should generalize broadly from a training stimulus to other stimuli capable of supporting learning, whereas the alternatives predict that learning should be relatively specific to the training stimulus. Here we show that deep nucleus plasticity, as assessed by conditioned eyelid responses produced without input from the cerebellar cortex, is relatively specific to the training conditioned stimulus (CS). We trained rabbits to a tone or light CS with periorbital stimulation as the unconditioned stimulus (US), and pharmacologically disconnected the cerebellar cortex during a posttraining generalization test. The short-latency conditioned responses unmasked by this treatment showed strong decrement along the dimension of auditory frequency and did not generalize across stimulus modalities. These results cannot be explained solely by a cell-wide increase in the excitability of deep nucleus neurons, and imply that an input-specific mechanism in the deep cerebellar nucleus operates as well.


Relation of immunoreactive gastric inhibitory polypeptide to changes in glycaemic control and B cell function in type 1 (insulin-dependent) diabetes mellitus.
The effect of strict glycaemic control on plasma immunoreactive gastric inhibitory polypeptide (IR-GIP) concentrations and pancreatic B cell function as estimated by plasma C-peptide was evaluated in 14 Type 1 (insulin-dependent) diabetics. The effect was estimated by giving a test meal before (test 1) and after (test 2) 1 week with near normal blood glucose control (mean blood glucose 6.7 +/- 0.2 mmol/l) and again 3 weeks later (test 3) in the outpatient clinic. The glycaemic control was significantly improved at test 2 and test 3 compared with that of test 1. The IR-GIP concentrations before and after the meals were similar at all three tests and not different from those found in 21 normal controls. In 8 patients with a significant B cell response at test 1, B cell function was significantly improved both at test 2 and test 3 but no change in fasting or post-prandial IR-GIP concentrations was found and no correlation between B cell function and IR-GIP existed. We conclude that strict glycaemic control improves B cell function but does not modulate plasma IR-GIP concentrations. Factors other than GIP seem to be of greater importance in determining the magnitude of B cell function in Type 1 diabetes.


Genetics and Intervention Research.
As part of this special section on genetics and behavioral intervention, we discuss the articles by McGue, Osler, and Christensen (2010, this issue) and by Davey Smith (2010, this issue). In the second half of the present article, we consider the integration of genetics and intervention research more broadly. The two articles describe ways to use genetic controls to infer causation from correlational ("observational") data without intervention. McGue et al. discuss the use of twins discordant for exposure, which is a variant of the cotwin control method. This method can show that the link between an exposure and outcome is not entirely mediated genetically. Davey Smith discusses a method called Mendelian randomization that uses DNA to draw causal inferences without the need for experimental intervention. Despite the possibilities for using genetic controls to infer causation from correlational data in order to attenuate the need for intervention studies, we are most excited about the opportunities for integrating genetics and intervention research, especially as new DNA technologies make it possible to incorporate genetics in any intervention research.


On the heterogeneity of native ribosomal subunits in Ehrlich-ascites-tumor cells cultured in vitro.
Native small ribosomal subunits in cultured Ehrlich ascites tumor cells, analyzed by high-resolution CsCl isopycnic centrifugation, consist of at least five classes of particles. These particles with buoyant densities of 1.39, 1.42, 1.45, and 1.51 g/cm3 were designated as SI, SII, SIII, SIV and SV, respectively. They were different from the ribosome-derived 40-S subunits which have a density of 1.54 g/cm3. Native large ribosomal subunits consist of at least two classes of particles with densities of 1.57 g/cm3 (LI) and 1.59 g/cm3 (LII), respectively. The ribosome-derived 60-S subunits have the same buoyant density as LII particles. Labeling of Ehrlich ascites cells with radioactive uridine and a subsequent chase in the presence of RNA-synthesis inhibitors shows that radioactivity was incorporated first in precursor particles with a density of 1.48 g/cm3 and then subsequently appeared in SIII and SII particles. Met-tRNAf is found exclusively on native 40-S particles with densities of 1.42 and 1.49 g/cm3. This result has been observed in cells labeled with [35S] methionine in vivo as well as with tRNA charged in vitro. The possibility that SII particles contain 40-S initiation complexes is discussed.


[Concerted action program for quality assurance of European community hospitals as applied in Israel].
The concerted action program on quality assurance (QA) of the European Community (COMAC) was implemented in 14 hospitals in Israel. It was focused on prevention of pressure sores, preoperative assessment, prophylactic use of antibiotics in elective surgery and proper keeping of medical and nursing records. Mechanisms for quality assessment and interventions for improvement and monitoring were implemented. Improvement in QA infrastructure, processes of care in the project topics, and in QA awareness were found. Management involvement, previous motivation for professional improvement, behavioral strategies for change and having highly respected QA leaders were found to contribute to the development of effective QA. Professional associations and national authorities should be responsible for establishing support systems as well as promoting legislation, formulating guidelines, recommending allotment of resources, and developing educational programs.


Autogenous bone grafts in oral implantology-is it still a "gold standard"? A consecutive review of 279 patients with 456 clinical procedures.
This study assessed the clinical outcomes of graft success rate and early implant survival rate after preprosthetic alveolar ridge reconstruction with autologous bone grafts. A consecutive retrospective study was conducted on all patients who were treated at the military outpatient clinic of the Department of Oral and Plastic Maxillofacial Surgery at the military hospital in Ulm (Germany) in the years of 2009 until 2011 with autologous bone transplantation prior to secondary implant insertion. Intraoral donor sites (crista zygomatico-alveolaris, ramus mandible, symphysis mandible, and anterior sinus wall) and extraoral donor site (iliac crest) were used. A total of 279 patients underwent after a healing period of 3-5 months routinely computer tomography scans followed by virtual implant planning. The implants were inserted using guided oral implantation as described by Naziri et al. All records of all the consecutive patients were reviewed according to patient age, history of periodontitis, smoking status, jaw area and dental situation, augmentation method, intra- and postoperative surgical complications, and surgeon's qualifications. Evaluated was the augmentation surgical outcome regarding bone graft loss and early implant loss postoperatively at the time of prosthodontic restauration as well a follow-up period of 2 years after loading. A total of 279 patients underwent 456 autologous augmentation procedures in 546 edentulous areas. One hundred thirteen crista zygomatico-alveolaris grafts, 104 ramus mandible grafts, 11 symphysis grafts, 116 grafts from the anterior superior iliac crest, and 112 sinus lift augmentations with bone scrapes from the anterior facial wall had been performed. There was no drop out or loss of follow-up of any case that had been treated in our clinical center in this 3-year period. Four hundred thirty-six (95.6%) of the bone grafts healed successfully, and 20 grafts (4.4%) in 20 patients had been lost. Fourteen out of 20 patients with total graft failure were secondarily re-augmented, and six patients wished no further harvesting procedure. In the six patients, a partial graft resorption was detected at the time of implantation and additional simultaneous augmentation during implant insertion was necessary. No long-term nerve injury occurred. Five hundred twenty-five out of 546 initially planned implants in 259 patients could be inserted into successfully augmented areas, whereas 21 implants in 20 patients due to graft loss could not be inserted. A final rehabilitation as preplanned with dental implants was possible in 273 of the 279 patients. The early implant failure rate was 0.38% concerning two out of the 525 inserted implants which had to be removed before the prosthodontic restoration. Two implants after iliac crest augmentation were lost within a period of 2 years after loading, concerning a total implant survival rate after 2 years of occlusal loading rate of 99.6% after autologous bone augmentation prior to implant insertion. This review demonstrates the predictability of autologous bone material in alveolar ridge reconstructions prior to implant insertion, independent from donor and recipient site including even autologous bone chips for sinus elevation. Due to the low harvesting morbidity of autologous bone grafts, the clinical results of our study indicate that autologous bone grafts still remain the "gold standard" in alveolar ridge augmentation prior to oral implantation.


Complications following resection of the olecranon bursa.
We retrospectively reviewed 37 cases of resection of the olecranon bursa and noted wound healing problems in 10 (27%) and recurrence in 8 (22%). A lateral arm flap was necessary in one patient. Conservative treatment remains the treatment of choice for olecranon bursitis. Differentiation between septic and non-septic cases is challenging. The risk of wound healing problems and recurrence should be taken into account when planning surgical resection.


Species delimitation for the molecular taxonomy and ecology of the widely distributed microbial eukaryote genus Euplotes (Alveolata, Ciliophora).
Recent advances in high-throughput sequencing and metabarcoding technologies are revolutionizing our understanding of the diversity and ecology of microbial eukaryotes (protists). The interpretation of protist diversity and the elucidation of their ecosystem function are, however, impeded by problems with species delimitation, especially as it applies to molecular taxonomy. Here, using the ciliate Euplotes as an example, we describe approaches for species delimitation based on integrative taxonomy by using evolutionary and ecological perspectives and selecting the most appropriate metabarcoding gene markers as proxies for species units. Our analyses show that: Euplotes (sensu lato) comprises six distinct clades, mainly as result of ecological speciation; the validity of the genera Euplotes (sensu stricto), Euplotoides, Euplotopsis and Moneuplotes are not supported; the vannus-type group, which includes species without distinct morphological differences, seems to be undergoing incipient speciation and contains cryptic species; the hypervariable V4 region of the small subunit rDNA and D1-D2 region of the large subunit rDNA are the promising candidates for general species delimitation in Euplotes.


Severe pulmonary regurgitation mimicking constrictive pericarditis: a case report-the sac or the content? That is the question.
Constrictive pericarditis (CP) is a pathological condition of the pericardium, resulting from fibrosis, scarring, and calcification of the pericardium. Other conditions have been reported to mimic 'constrictive physiology' despite the presence of an intact pericardium. However, there has been no report of pulmonary regurgitation (PR) mimicking the haemodynamic characteristics of CP. A 51-year-old woman was admitted to our institute because of severe right-sided heart failure. Transthoracic echocardiography revealed severe PR concomitant with significant dilatation of the right-sided heart. Septal bounce and the respiratory reciprocation of the transmitral and transtricuspid inflow velocities were also observed, indicating exacerbated ventricular interdependence. Cardiac catheter examination demonstrated elevated right atrial pressure with a prominent y descent, dip, and plateau waveform in the right ventricular pressure, and equalization of the diastolic pressure of all cardiac chambers, which are quite consistent with CP. On surgical inspection, however, there was no pericardial thickening or adhesion, indicating no obvious signs of CP. Pericardial constriction results from the relative relationship between intrapericardial volume and pericardial reserve. When the intrapericardial volume exceeds the physiological limit, the cardiac chambers compete with each other in a fixed pericardial space. In this case, prominent dilation of the right-sided chambers caused by severe PR resulted in overstretching of the pericardium above the pericardial reserve, which led to a characteristic haemodynamic picture that resembled CP. Thus, it is important to recognize the diagnostic pitfall in the preoperative evaluation of a 'CP mimic physiology'.


Intermediate filament cytoskeletal proteins associated with bovine lens native membrane fractions.
To examine the intermediate filament cytoskeletal proteins associated with native membrane fractions isolated from bovine lenses. Decapsulated bovine lenses were divided into cortex and nucleus. The lens regions were homogenized and separated into water-soluble and water-insoluble fractions by centrifugation. Sedimenting membrane fractions were isolated from the water-insoluble fraction by discontinuous sucrose-density-gradient centrifugation and the non-sedimenting membrane fractions were isolated from the Kbr high-density water-soluble fractions by flotation, during overnight centrifugation. The intermediate filament peptides of the membrane fractions were examined by Western blot analysis, using monoclonal antibodies to filensin, cytoskeletal protein 49 (CP49) and vimentin. Filensin immunoreactive peptides were found in all membrane fractions of both cortex and nucleus. The parent 115 kDa filensin was found almost exclusively in the urea-soluble protein of cortical membrane fractions, and was the predominant filensin immunoreactive peptide only in the urea-soluble protein of the cortical sedimenting membrane fraction isolated from the 25%/45% sucrose density interface. The predominant filensin immunoreactive peptide of all other samples migrated with a M(r) of 53 kDa. CP49 immunoreactive peptides were found almost exclusively in the urea-soluble protein of all membrane fractions from both the cortex and nucleus. The cortical non-sedimenting membrane fraction and the nuclear membrane fraction of the 25%/45% sucrose density interface were notably deficient in CP49. Vimentin immunoreactive peptides were found in both urea-soluble and urea-insoluble proteins of membrane fractions from the cortex only. Vimentin was particularly enriched in the cortical non-sedimenting membrane fraction. The urea-insoluble filensin immunoreactive peptides were only partially removed by alkali extraction, indicating a very avid association with the membrane. Two dimensional electrophoresis revealed that the urea-soluble protein of the major cortical membrane fraction contained two different filensin-derived 53 kDa fragments. The non-sedimenting membrane fraction, which may reflect a distinct domain of the lens plasma membrane, possesses a membrane-associated cytoskeletal composition different from that of the major sedimenting membrane fractions.


Geometrical control of interface patterning underlies active matter invasion.
Interaction between active materials and the boundaries of geometrical confinement is key to many emergent phenomena in active systems. For living active matter consisting of animal cells or motile bacteria, the confinement boundary is often a deformable interface, and it has been unclear how activity-induced interface dynamics might lead to morphogenesis and pattern formation. Here, we studied the evolution of bacterial active matter confined by a deformable boundary. We found that an ordered morphological pattern emerged at the interface characterized by periodically spaced interfacial protrusions; behind the interfacial protrusions, bacterial swimmers self-organized into multicellular clusters displaying +1/2 nematic defects. Subsequently, a hierarchical sequence of transitions from interfacial protrusions to creeping branches allowed the bacterial active drop to rapidly invade surrounding space with a striking self-similar branch pattern. We found that this interface patterning is geometrically controlled by the local curvature of the interface, a phenomenon we denote as collective curvature sensing. Using a continuum active model, we revealed that the collective curvature sensing arises from enhanced active stresses near high-curvature regions, with the active length scale setting the characteristic distance between the interfacial protrusions. Our findings reveal a protrusion-to-branch transition as a unique mode of active matter invasion and suggest a strategy to engineer pattern formation of active materials.


Folate-Modified Poly(malic acid) Graft Polymeric Nanoparticles for Targeted Delivery of Doxorubicin: Synthesis, Characterization and Folate Receptor Expressed Cell Specificity.
A novel amphiphilic biodegradable cholesterol and poly(ethylene glycol)-folate grafted poly(α,β-malic acid) (PMA-g-Chol/PEG-FA) was synthesized and characterized as self-assembled nanoparticles for targeted delivery of doxorubicin (DOX). The nanoparticles showed extremely low critical aggregation concentrations (CAC), appropriate zeta potential, narrow size distribution, good stability in serum conditions and negligible toxicity. After encapsulation'of DOX, PMA-g-Chol/PEG-FA nanoparticles showed significantly reduced cell viability (up to 30% for Hela and 27% for 4T1 cells) compared with the non-targeted ones on carcinoma cells with different levels of folate receptor (FR) expression. While no difference was detected on HEK293 cells (FR receptor negative) between the two nanoparticles. Addition of extra free folate obviously decreased the cellular mortality and inhibited the cellular uptake of targeted nanoparticles. In the Hela/HEK293 co-culture model, folate conjugated nanoparticles showed specific affiliation with Hela cells other than HEK293 cells, indicating good targeting property of the delivery system. As detected from ex vivo fluorescent imaging, PMA-g-Chol/PEG-FA nanoparticles could accumulate at tumor site with higher selectivity compared to PMA-g-Chol/PEG nanoparticles and DOX x HCl. In vivo antitumor studies confirmed the significant tumor inhibition efficacy of drug-loaded PMA-g-Chol/PEG-FA nanoparticles with lower toxicity to normal tissues than DOX x HCI at the same dosage.


Pilot-scale removal of pharmaceuticals in municipal wastewater: Comparison of granular and powdered activated carbon treatment at three wastewater treatment plants.
Adsorption with activated carbon is widely suggested as an option for the removal of organic micropollutants including pharmaceutically active compounds (PhACs) in wastewater. In this study adsorption with granular activated carbon (GAC) and powdered activated carbon (PAC) was analyzed and compared in parallel operation at three Swedish wastewater treatment plants with the goal to achieve a 95% PhAC removal. Initially, mapping of the prevalence of over 100 substances was performed at each plant and due to low concentrations a final 22 were selected for further evaluation. These include carbamazepine, clarithromycin and diclofenac, which currently are discussed for regulation internationally. A number of commercially available activated carbon products were initially screened using effluent wastewater. Of these, a reduced set was selected based on adsorption characteristics and cost. Experiments designed with the selected carbons in pilot-scale showed that most products could indeed remove PhACs to the target level, both on total and individual basis. In a setup using internal recirculation the PAC system achieved a 95% removal applying a fresh dose of 15-20 mg/L, while carbon usage rates for the GAC application were much broader and ranged from <28 to 230 mg/L depending on the carbon product. The performance of the PAC products generally gave better results for individual PhACs in regards to carbon availability. All carbon products showed a specific adsorption for a specific PhAC meaning that knowledge of the target pollutants must be acquired before successful design of a treatment system. In spite of different configurations and operating conditions of the different wastewater treatment plants no considerable differences regarding pharmaceutical removal were observed.


Determination of brain death in children--a medicial center experience.
We retrospectively reviewed our organ donation team's database (timeframe: 1988-2003) identifying 18 patients under 18 years old meeting criteria for the diagnosis of brain death. We emphasized the primary diagnosis, observation period, physical findings, neurological findings, and the apnea test results according to the 1987 guidelines for brain death determination. The average age at diagnosis was 13 years and 8 months old (range: 3 months-18 years; median: 16 years). Fifteen children (82%) suffered head injuries; one (6%) had hypoxic ischemic encephalopathy; one (6%) suffered intracranial hemorrhage; and one (6%) had a brain tumor. The average observation period from clinical suspicion of brain death to the brain death test was 146 hours (range: 16-1416 hours; median: 72 hours). Among the 18 patients fulfilling the criteria, brain death was declared after two successful apnea tests performed at least 4 hours apart (average: 6 hours; range: 4-18 hours). Solid organ procurement was successful in all patients (100%). Use of the guidelines for determining brain death in children is limited in Taiwan; however, we found that the previously used guidelines in Taiwan might be appropriate for children above 3 years old, although further study is necessary to determine the roles of electroencephalography and cerebral blood flow measurement in younger children.


Clinical significance of iodine-123-15-(p-iodophenyl)-3-R, S-methylpentadecanoic acid myocardial scintigraphy in patients with aortic valve disease.
The present study sought to determine whether myocardial fatty acid metabolism as assessed with iodine-123-labeled 15-(p-iodophenyl)-3-R,S-methylpentadecanoic acid (BMIPP) scintigraphy is impaired in patients with aortic valve disease (AVD) and whether the degree of the metabolic abnormality reflects the severity of AVD. BMIPP scintigraphy was performed in 12 patients with aortic stenosis (AS), 14 patients with aortic regurgitation (AR), and 9 healthy volunteers, and from that the heart-mediastinum uptake ratio (H/M ratio) corrected by the left ventricular (LV) mass (U/Mass ratio) and the myocardial washout rate (WR) were obtained. The H/M ratio tended to be higher in patients than in healthy volunteers (3.3 +/- 0.7 for AS, 3.5 +/- 0.5 for AR, 3.0 +/- 0.3 for healthy volunteers), and the WR was significantly higher in patients than in healthy volunteers (42.8 +/- 9.1% for AS, 35.7 +/- 6.5% for AR, 19.6 +/- 9.1% for healthy volunteers, p<0.01). In the AS patients, the U/Mass ratio showed significant negative correlations (r=-0.79 to -0.90, all p<0.01) and the WR showed significant positive correlations (r=0.61 to 0.82, all p<0.01) with transaortic pressure gradient, LV wall thickness, and LV mass. Similarly, in AR patients these BMIPP parameters showed proportional changes to the LV volumes and LV mass (r=-0.79 to -0.83, all p<0.01 for U/Mass ratio, r=0.55 to 0.70, p<0.05 to <0.01 for WR). In the 9 patients who underwent aortic valve replacement, the BMIPP parameters tended to normalize with increasing U/Mass ratio (0.90 +/- 0.41 x 10(-2)/g to 1.34 +/- 0.59 x 10(-2)/g, p<0.05) and decreasing WR (41.9 +/- 8.8% to 35.4 +/- 9.2%, p<0.01) after surgery. Myocardial fatty acid metabolism as assessed with BMIPP scintigraphy was impaired in patients with aortic valve disease and the U/Mass ratio and WR reflect the severity. These parameters may be useful for the noninvasive assessment of the myocardial metabolic abnormalities caused by hemodynamic overload.


Construction of multiple-epitope tag sequence by PCR for sensitive Western blot analysis.
Epitope tagging is a powerful technique to characterize a recombinantly expressed protein encoded by cDNA without the purification of the protein and the immunization of animals. In some cases, however, the expression of a tagged protein is too low to analyze by Western blot. We have developed a simple method to generate tandem repetitive nucleotide sequence by PCR, which allows us to label a protein of interest with a multiple-epitope tag. When five myc epitopes were attached to vaccinia virus protein CrmA, its signal was multiplied 5.8 times in Western blot analysis, compared with that of one epitope-tagged CrmA.


Effect of brief exercise on circulating insulin-like growth factor I.
An acute insulin-like growth factor I (IGF-I) response to 10 min of above-lactate threshold cycle ergometer exercise was studied in 10 subjects (age 22-35 yr). Each subject exercised on three separate mornings after ingesting one of two isocaloric isovolemic liquid meals high in either fat or glucose or an isovolemic noncaloric placebo. The high-fat meal attenuated the growth hormone (GH) response (Cappon et al., J. Clin. Endocrinol. Metab. 76: 1418-1422, 1993). In contrast, IGF-I increased equally for all protocols [e.g., after the placebo meal IGF-I increased from 21,716 (SE) ng/ml preexercise to 25,316 ng/ml at 10 min of exercise; P < 0.05]. IGF-I peaked by the 10th min of exercise, like GH, and remained significantly elevated for only 20 min of recovery. We tested for possible GH-dependent mechanisms in which circulating IGF-I would increase 12-24 h after exercise. Ten subjects (age 23-32 yr) performed 10 min of above-lactate threshold exercise at 9, 10, and 11 A.M. GH was elevated after the first exercise bout (peak GH 6.05 +/- 1.45 ng/ml; P < 0.001) but was significantly reduced for the second and third bouts (peak GH 2.52 +/- 0.76 and 1.50 +/- 0.40 ng/ml, respectively). No increase in IGF-I was observed by 8 A.M. on the following day. Heavy ergometer exercise led to brief and small increases in circulating IGF-I that were independent of circulating GH.(ABSTRACT TRUNCATED AT 250 WORDS)


Influence of lockdown during the COVID-19 pandemic on use of pediatric emergency health services: A retrospective observational study from a Tertiary Care Hospital in New Delhi.
January 30, 2020, marked the beginning of the COVID-19 pandemic in India. Various emergency measures were taken to contain the spread of COVID-19 including extended periods of complete lockdown. The impact of these measures on routine and emergency health services was unforeseen. Hence, we conducted this study to critically analyze the effects of restrictions imposed during the COVID-19 pandemic (including lockdown) on the utilization of health services, especially emergency services. We compared patient's attendance in the outpatient department and pediatric emergency department (PED) and changes in clinicepidemiological profiles (before and during COVID-19) in a tertiary care hospital. We observed a 43% decline in PED visits which decreased to 75% during the period of strict lockdown (P = 0.005). Reduction in emergency department visits was noticed uniformly in all disease categories. This study highlights the urgent need to plan for robust health-care support system for the delivery of preventive and curative services to vulnerable age groups during any emergency.


Downstaging cancer in rural Africa.
Cancer is usually diagnosed late in rural Africa leading to incurability and abbreviated survival. Many curable cancers present on the body surface, often recognizable early by laymen as suspicious, justifying professional referral. Cancer diagnoses in two randomly chosen Tanzanian villages were compared after conventional dispensary self-referral vs. proactive visits in the home. Village navigators organized trips for professional consultation. In the control village 21% were self-referred, 20% of them were sent on as suspicious, 78% had cancer (8% in men) 0.9% of the village population. In the intervention village 99% were screened, 14% were referred for professional opinion, 93% had cancer (32% in men) 1.6% (p < 0.01 compared with control village). In the second and third years similar activity yielded 0.5% cancer annually in the control village for a 3 year total of 1.86% whereas interventional villagers had 1.4% and 0.6% cancer for a 3 year total of 3.56% (p < 0.001). Downstaging was recognized in the second and third years of intervention from 23 to 51 to 74% Stages I and II (p < 0.001) but in the control village Stages I and II changed from 11% to 22% to 37% (p = NS). The greatest downstaging occurred in breast and cervix cancers.


[Clinical management of a patient with alcoholic cirrhosis].
Prolonged abstinence from alcohol is crucial in the management and prognosis of a patient with alcoholic cirrhosis. It is also important to prevent complications such as variceal bleeding, hepatocellular and extrahepatic cancers, and malnutrition. Liver transplantation should be considered in patients with persistent liver failure in spite of complete cessation of alcohol consumption. We provide some recommendations in commonly encountered clinical situations for compensated and decompensated alcoholic cirrhosis.


Neurons cultured from developing rat brain attach and spread preferentially to laminin.
Neurons, mechanically dissociated from newborn rat brain and identified by immunostaining for neurofilaments, attached preferentially to laminin-coated coverslips without need of an underlying glial monolayer. The most extensive neurite outgrowth was seen when 20-30 micrograms/ml of laminin was used to coat the coverslips. Higher concentrations of laminin (greater than 30 micrograms/ml) supported single neurons to spread on the coverslips. Fibronectin coating of the coverslips allowed glial cells to attach more rapidly than on uncoated surfaces, but it did not support neuronal spreading or neurite outgrowth. Spreading of neurons and neurite outgrowth were completely inhibited by preincubation of laminin-coated coverslips with laminin antibodies but were unaffected by fibronectin antibodies. These results indicate that laminin is an attachment and spreading factor for central neurons in culture and suggest the presence of a laminin receptor on the neuronal cell surfaces.


[Effect of tumor necrosis factor-alpha on the permeability of strial capillary endothelial cells in guinea pig cochlea].
To perform an initial study on possible molecular regulatory mechanisms of TNF-alpha to the permeability of strial capillary endothelial cells in guinea pig cochlea Strial capillary endothelial cells in guinea pig cochlea was dissociated and cultured to establish a model for its permeability in vitro. The animals were divided into TNF-alpha, TNF-alpha + L-arginine (L-Arg), TNF-alpha + NG-monomethyl-L-arginine (L-NMMA) and control groups. In order to measure the alternations of permeation rate, Evans blue was used in different time by different concentrations to detect contents of F-actin in each group under immunofluorescence laser scanning confocal microscope. TNF-alpha increased permeability of strial capillary endothelial cells notably, and permeation rate to Evans blue heightened significantly (P < 0.01) and increased under higher concentrations of TNF-alpha as well as lengthened the durations. Being as a NO donator, L-Arg significantly enhanced the ability of TNF-alpha which increasing permeability of strial capillary endothelial cells; permeation rate to Evans blue heightened significantly (P < 0.01), which obviously rose up when the concentration of Evans blue added. L-NMMA, NO inhibitor, was able to weaken the ability of TNF-alpha which increasing permeability of strial capillary endothelial cells; permeation rate to Evans blue decreased significantly (P < 0.01), which obviously fell down when the concentration of L-NMMA added. TNF-alpha decreased F-actin in strial capillary endothelial cells (P < 0.01), more TNF-alpha, less F-actin; L-Arg could be further the inhibition to F-actin content (P < 0.05); L-NMMA held back the trend (P < 0.05). TNF-alpha can increase permeability of strial capillary endothelial cells of guinea pig cochlea. NO may be one of the important molecular regulators to permeability of strial capillary endothelial cells by TNF-alpha, and changes of F-actin content probably relate to the regulation in these cells.


Characterization of the developing small intestine in the absence of either GATA4 or GATA6.
Studies of adult mice lacking either GATA4 or GATA6 in the small intestine demonstrate roles for these factors in small intestinal biology. Deletion of Gata4 in the adult mouse intestine revealed an essential role for GATA4 in jejunal function. Deletion of Gata6 in the adult mouse ileum alters epithelial cell types and ileal enterocyte gene expression. The effect of deletion of Gata4 or Gata6 alone during embryonic small intestinal development, however, has not been examined. We recently demonstrated that loss of both factors in double conditional knockout embryos causes severe defects in jejunal development. Therefore, the goal of this study is to provide phenotypic analysis of the small intestine of single Gata4 and Gata6 conditional knockout embryos. Villin-Cre was used to delete Gata4 or Gata6 in the developing intestinal epithelium. Elimination of either GATA4 or GATA6 in the jejunum, where these factors are co-expressed, caused changes in enterocyte and enteroendocrine cell gene expression. Ectopic expression of markers of the ileal-specific bile acid metabolism pathway was induced in GATA4-deficient jejunum but not in GATA6-deficient jejunum. A subtle increase in goblet cells was also identified in jejunum of both mutants. In GATA6-deficient embryonic ileum, villus length was altered, and enterocyte gene expression was perturbed including ectopic expression of the colon marker Car1. Goblet cells were increased, and enteroendocrine cells were decreased. Overall, we show that aspects of the phenotypes observed in the small intestine of adult Gata4 and Gata6 conditional knockout mice emerge during development. The effect of eliminating GATA6 from the developing ileum was greater than that of eliminating either GATA4 or GATA6 from the developing jejunum likely reflecting functional redundancy between these factors in the jejunum. Although GATA4 and GATA6 functions overlap, our data also suggest unique functions for GATA4 and GATA6 within the developing intestine. GATA4 likely operates independently of GATA6 within the jejunum to regulate jejunal versus ileal enterocyte identity and consequently jejunal physiology. GATA6 likely regulates enteroendocrine cell differentiation cell autonomously whereas GATA4 affects this population indirectly.


Neural modulation of glucose-dependent insulinotropic peptide (GIP) and insulin secretion in conscious dogs.
Glucose-dependent insulinotropic peptide (GIP) is a potent incretin, but it remains unclear whether this effect is dependent upon intact vagal pathways. In four conscious dogs, plasma GIP, plasma insulin, and plasma glucose responses were measured after intraduodenal administration of a defined formula diet, after glucose was perfused intraduodenally, and after insulin-mediated hypoglycemia with and without bilateral cryogenic blockade of the cervical vagus nerves. Vagal blockade did not alter elevations of plasma GIP after the defined formula diet or after glucose. However, with the vagi blocked plasma insulin responses were suppressed after the enteral diet (-52 +/- 8%) and after intraduodenal glucose (-55 +/- 4%), without changes in plasma glucose. Intravenous atropine (50 micrograms/kg) did not modify the GIP responses to intraduodenal perfusions of the defined formula diet or to glucose, but did suppress plasma insulin responses to baseline values. Insulin hypoglycemia without or with vagal blockade had no effect on basal concentrations of plasma GIP. These results indicate that vagal muscarinic and nonvagal muscarinic pathways participate in the control of the intestinal phase of insulin secretion, but the regulation of GIP secretion is independent of vagal or muscarinic neural control mechanisms.


Electron-microscopic cytochemistry of the catecholaminergic innervation of ACTH-containing neurons in the rat hypothalamic arcuate nucleus.
The synaptic relationship between catecholamine terminals and adrenocorticotropic hormone (ACTH)-containing neurons in the arcuate nucleus (AN) of the rat hypothalamus was investigated by electron microscopy, using ACTH immunocytochemistry combined with autoradiography after 3H-dopamine (3H-DA) injection or 5-hydroxydopamine (5-OHDA) uptake in the same tissue section. ACTH-like (ACTH-LI) immunoreactive nerve cell bodies and fibers received synaptic inputs by axon terminals labeled with 3H-DA or 5-OHDA in the AN. This suggests that catecholaminergic neurons, at least DA- and 5-OHDA-containing neurons, may play an important role in the regulation of ACTH secretion or other functions of ACTH neurons via synapses in the AN of the rat hypothalamus.


In vitro induction of HLA-restricted cytotoxic T lymphocytes against autologous Epstein-Barr Virus transformed B lymphoblastoid cell line.
Cytotoxic T lymphocytes (CTL) against autologous EBV-transformed B lymphoblastoid cell line (LCL) were induced in vitro by culturing peripheral blood lymphocytes (PBL) of healthy donors together with mitomycin C-treated autologous LCL for 6 days. The cytotoxic cells developed only from the E-rosette-positive fraction but not from the negative fraction of PBL. These CTL killed autologous LCL but not PWM-stimulated autologous PBL. In addition, the CTL killed allogeneic LCL when at least 1 of the HLA-A antigens was identical with that of the LCL of CTL donor. However, identity of HLA-B and HLA-C antigens was not enough for a significant killing of allogeneic LCL. The specificity of the CTL was also confirmed by a cold target inhibition test. These results indicated that the CTL induced specifically recognized EBV-transformed cells with HLA restriction.


Alcohol consumption in France: production, consumption, morbidity and mortality, prevention and education in the last three decades.
This paper examines trends in the French consumption of alcohol over the last three decades, and includes data on the production of alcoholic beverages, the consumption of different types of alcoholic and non-alcoholic beverages, morbidity and mortality related to the consumption of alcohol, and a brief description of French preventive and educational measures in this area. In the last 15 years, the average alcohol consumption has ceased growing and even began to decrease. Data are given comparing consumption trends in several European countries, as well as more detailed data of alcohol consumption and related phenomena in France by region, by sex, by age and by socio-economic status (the last two categories for men only). Finally, a schema of the social dynamics of alcohol consumption is presented.


Cloning of a rat gene encoding the histo-blood group A enzyme. Tissue expression of the gene and of the A and B antigens.
The complete coding sequence of a BDIX rat gene homologous to the human ABO gene was determined. Identification of the exon-intron boundaries, obtained by comparison of the coding sequence with rat genomic sequences from data banks, revealed that the rat gene structure is identical to that of the human ABO gene. It localizes to rat chromosome 3 (q11-q12), a region homologous to human 9q34. Phylogenetic analysis of a set of sequences available for the various members of the same gene family confirmed that the rat sequence belongs to the ABO gene cluster. The cDNA was transfected in CHO cells already stably transfected with an alpha1,2fucosyltransferase in order to express H oligosaccharide acceptors. Analysis of the transfectants by flow cytometry indicated that A but not B epitopes were synthesized. Direct assay of the enzyme activity using 2' fucosyllactose as acceptor confirmed the strong UDP-GalNAc:Fucalpha1,2GalalphaGalNAc transferase (Atransferase) activity of the enzyme product and allowed detection of a small UDP-Gal:Fucalpha1,2GalalphaGal transferase (B transferase) activity. The presence of the mRNA and of the A and B antigens was searched in various BDIX rat tissues. There was a general good concordance between the presence of the mRNA and that of the A antigen. Tissue distributions of the A and B antigens in the homozygous BDIX rat strain were largely different, indicating that these antigens cannot be synthesized by alleles of the same gene in this rat inbred strain.


Profiling of cell cycle genes of breast cells exposed to etodolac.
Breast cancer represents the second leading cause of cancer-related deaths in the world. There is increasing evidence that perturbation of cell cycle regulation is an important contributing factor to various cancer progression stages. There are key checkpoints in the cell cycle involving various regulatory proteins. The relationship between these cell cycle regulatory proteins and cell cycle arrest by cyclo-oxygenase (COX) inhibitors during neoplastic progression remains largely unknown. Preclinical studies and epidemiological investigations have consistently shown that non-steroidal anti-inflammatory drugs have some anti-proliferative and anti-oxidative stress response on various tumors. In this study, the effect of etodolac, a 1,8-diethyl-1,3,4,9-tetrahydro-pyrano (3,4-beta) indole-1-acetic acid on signaling pathways was investigated by examining the differential expression of various cell cycle regulatory protein genes. A human cell cycle gene array was used to profile the expression of 96 genes involved in the cell cycle regulation. Differentially expressed genes were highly altered by etodolac treatment. Twenty-six genes were up- and 20 down-regulated with 0.5 and 2 mM etodolac treatment, respectively. Seven genes (ATM, BAX, CCNA2, CDC27, RAD50 and p21) were prominently altered, and six (ATM, CCND2, CCNF, CDC20, CDK1A and RAD50) were commonly altered with both concentrations. This finding indicated that etodolac could play a critical role on cancer cells by inducing cell death.


Neonatal Lemierre Syndrome: Youngest Reported Case and Literature Review.
A previously healthy 5-week-old female was admitted for sepsis secondary to methicillin-resistant Staphylococcus aureus (MRSA) bacteremia. After several days of hospitalization, she experienced acute decompensation in mental status despite having received targeted antibiotic therapy. Imaging revealed left peritonsillar/parapharyngeal space abscess, left venous thrombophlebitis of the internal jugular vein, and septic emboli of the lungs and brain consistent with Lemierre syndrome. Bedside needle aspiration of the parapharyngeal abscess confirmed MRSA involvement. Unfortunately, the patient continued to deteriorate over the next several days and life support was withdrawn on hospital day 16. We present the youngest reported case of Lemierre syndrome and review the literature.


Corticosteroids stimulate the differentiation of growth hormone cells but suppress that of prolactin cells in the fetal rat pituitary.
An organ culture study was carried out to examine the effects of adrenal corticosteroids on the development of growth hormone (GH) cells and prolactin (PRL) cells. The adenohypophysial primordia were separated from 13.5-day-old fetal rats and maintained in vitro for 8 days with or without cortisol. Immunohistochemical examination of these explants showed that cortisol stimulated the differentiation of GH cells but suppressed that of PRL cells in a dose-dependent manner. In the absence of cortisol there were more PRL cells. Corticosterone had a similar effect on the developing adenohypophysis. When the pituitary primordia of Day 16.5 were cultured for 5 days and studied by the in situ hybridization technique, the expression of GH and PRL mRNA was found to be parallel with the immunoreactivity of the respective hormones. These data are discussed in relation to the normal development of GH and PRL cells in the fetal rat adenohypophysis.


Pelvic floor dysfunction, and effects of pregnancy and mode of delivery on pelvic floor.
Pelvic floor dysfunction (PFD), although seems to be simple, is a complex process that develops secondary to multifactorial factors. The incidence of PFD is increasing with increasing life expectancy. PFD is a term that refers to a broad range of clinical scenarios, including lower urinary tract excretory and defecation disorders, such as urinary and anal incontinence, overactive bladder, and pelvic organ prolapse, as well as sexual disorders. It is a financial burden on the health care system and disrupts women's quality of life. Strategies applied to decrease PFD are focused on the course of pregnancy, mode and management of delivery, and pelvic exercise methods. Many studies in the literature define traumatic birth, usage of forceps, length of the second stage of delivery, and sphincter damage as modifiable risk factors for PFD. Maternal age, fetal position, and fetal head circumference are nonmodifiable risk factors. Although numerous studies show that vaginal delivery affects pelvic floor structures and their functions in a negative way, there is not enough scientific evidence to recommend elective cesarean delivery in order to prevent development of PFD. PFD is a heterogeneous pathological condition, and the effects of pregnancy, vaginal delivery, cesarean delivery, and possible risk factors of PFD may be different from each other. Observational studies have identified certain obstetrical exposures as risk factors for pelvic floor disorders. These factors often coexist; therefore, the isolated effects of these variables on the pelvic floor are difficult to study. The routine use of episiotomy for many years in order to prevent PFD is not recommended anymore; episiotomy should be used in selected cases, and the mediolateral procedures should be used if needed.


CHROMOSOMAL POLYMORPHISM IN ISOFEMALE LINES AND CAGE POPULATIONS OF DROSOPHILA MELANOGASTER.
Drosophila melanogaster populations in nature usually carry inversion polymorphisms. When they were transferred to and maintained in the laboratory as large cage populations, frequencies of polymorphic inversions were drastically decreased and finally eliminated. This "cage effect" was observed irrespective of the geographical origin of the population or the initial frequency of each inversion. The decrease and elimination of inversions in the cage was not overcome by changing conditions such as medium, temperature, or the number of isofemale lines (40-600) introduced. On the other hand, in the sets of isofemale lines derived from the same geographical origins as the cage populations, each of which was maintained as a small vial population, the inversion frequencies, though decreased from the initial frequencies, were kept at significantly high levels. The cage populations initiated with one or two isofemale lines also maintained the inversion polymorphisms that were as high as vial populations.


An individualized course in clinical psychiatry.
Although a variety of new teaching strategies and materials are available in education today, medical education has been slow to move away from the traditional lecture format. An individualized course in clinical psychiatry is described, both from a philosophical and operational frame of reference. Features of the course of include programmed instruction, behavioral objectives, criterion-referenced mastery level evaluations, contracting for grades, and utilization of small-group discussions. Attention is directed to the advantages of the new methods as well as to new problems which surfaced.


Keeping up with 'Mrs Chase': an analysis of nursing skill-learning.
Psychomotor skill-learning is a basic aspect of nursing education. Students are expected to perform skills in relation to patient care. How can nursing instructors optimize skill learning and how can they analyse where a student is going wrong? The critical components of the psychomotor-skill learning process are identified and listed under the control of the instructor or the student. A checklist provides for mutual examination of the process to see if a problem area can be isolated. The article provides a review of a process which tends to become second nature to experienced nurses, but it remains one of which nursing instructors must be conscious, in order to understand why some students do not learn nursing skills easily or safely.


Expression of matrix metalloproteinase 2 (MMP-2), E-cadherin and Ki-67 in metastatic and non-metastatic canine mammary carcinomas.
The aim of the study was to demonstrate the immunohistochemical expression of proteins that affect the metastatic potential of a tumour, including matrix metalloproteinase 2 (MMP-2) and E-cadherin. Another objective was to determine their correlation with the expression of the Ki-67 antigen in metastasizing and non-metastasizing mammary carcinomas in female dogs. The study was conducted on 32 canine mammary carcinomas (12 metastatic and 20 non-metastatic), classified as simple tubular and tubulopapillary carcinomas. Immunohistochemistry was performed to evaluate the expression of MMP-2, E-cadherin and Ki-67 antigen. MMP-2 was expressed in 85 % of the non-metastatic tumours and in all the metastatic tumours, while E-cadherin was expressed in 85 % of the non-metastatic tumours and in 66 % of the metastatic tumours. The Ki-67 antigen was expressed in 65 % of the non-metastatic tumours and in 91 % of the metastatic tumours. The mean Ki-67 expression was slightly higher in tumours that had metastasized (1.5 ± 0.90 vs 1.1 ± 0.94; p = 0.22). A similar relationship was found in terms of the intensity of the MMP-2 expression (2.9 ± 1.9 vs 2.7 ± 2.4; p = 0.50). A decrease in the expression of E-cadherin (2.8 ± 2.5) was found in metastatic tumours compared to the expression in non-metastatic tumours (3.2 ± 2.3). However, these differences were not statistically significant (p = 0.63). We did not show significant differences in MMP-2, E-cadherin and Ki-67 expression between metastatic and non-metastatic tumours due to low number of cases studied, however further experiments are necessary to assess the role of these antigens in the process of canine mammary tumours metastasis.


Peritrophic membrane of the penaeid shrimp Sicyonia ingentis: structure, formation, and permeability.
Peritrophic membranes (PTMs) are secreted acellular layers that separate ingested materials from the gut epithelium in a variety of invertebrates. In insects and crustaceans, PTMs are produced in the midgut trunk (MGT, or intestine), but the MGT in decapod crustaceans, unlike that of insects, is not involved with digestion or absorption of food. We demonstrate that the PTM in the penaeid shrimp Sicyonia ingentis is similar to that in other crustaceans that have been studied and is primarily composed of chitin. The lectin WGA binds only to the PTM and glycocalyx along the microvilli of the midgut cells, which is consistent with the suggestion that the chitin is synthesized along the microvilli. The PTM is only permeable to inert particles smaller than 20 nm. We also describe the secretion of granules, which fill the apices of the epithelial cells, into the ectoperitrophic space. Although their function is not clear, they do not contribute to the PTM.


A common immunological epitope existing between HIV-1 gp41 and human interferon-alpha and -beta.
Based on the similar effects that HIV-1 gp41 like human type I interferons (IFN-alpha and -beta) upregulated MHC expression and inhibited lymphocyte proliferation, we compared amino acid sequences of gp41 and human type I interferons (IFN-alpha and -beta), and found sequence-similarity existing between the immunosuppressive domain (aa583-599, LQARILAVERYLKDQQL) of HIV-1 gp41 and human IFN-alpha (region aa117-129) and IFN-beta (region aa128-134); besides, the immunosuppressive domain of HIV-1 gp41 shows sequence-homology with bovine and murine IFN-alpha and IFN-beta in the same region. We examined polyclonal antibodies to human IFN-alpha and IFN-beta respectively and found that both antibodies could recognize rsgp41 from two different sources. The polyclonal antibody to IFN-beta could bind to the immunosuppressive peptide (ISP, aa583-599, LQARILAVERYLKDQQL) of HIV-1 gp41, only when the ISP was coupled with carrier-protein, and inhibit, if preincubated with rsgp41, binding of rsgp41 to human H9 (T cells), Raji (B cells), and U937 (monocyte cells) completely. The polyclonal antibody to IFN-alpha could partially inhibit the binding of rsgp41 to U937 and Raji. These results indicate that a common immunological epitope exists between HIV-1 gp41 and human type I interferons.


Characterization of an improved donor fluorescent protein for Forster resonance energy transfer microscopy.
The genetically encoded fluorescent proteins (FP), used in combination with Forster resonance energy transfer (FRET) microscopy, provide the tools necessary for the direct visualization of protein interactions inside living cells. Typically, the Cerulean and Venus variants of the cyan and yellow FPs are used for FRET studies, but there are limitations to their use. Here, Cerulean and the newly developed monomeric Teal FP (mTFP) are compared as FRET donors for Venus using spectral and fluorescence lifetime measurements from living cells. The results demonstrate that when compared to Cerulean, mTFP has increased brightness, optimal excitation using the standard 458-nm laser line, increased photostability, and improved spectral overlap with Venus. In addition, the two-photon excitation and fluorescence lifetime characteristics are determined for mTFP. Together, these measurements indicate that mTFP is an excellent donor fluorophore for FRET studies, and that its use may improve the detection of interactions involving proteins that are difficult to express, or that need to be produced at low levels in cells.


Risk Factors for Small Bowel Obstruction After Laparoscopic Ileal Pouch-Anal Anastomosis for Inflammatory Bowel Disease: A Multivariate Analysis in Four Expert Centres in Europe.
Although laparoscopy is associated with a reduction in adhesions, no data are available about the risk factors for small bowel obstruction [SBO] after laparoscopic ileal pouch-anal anastomosis [IPAA]. Our aims here were to identify the risk factors for SBO after laparoscopic IPAA for inflammatory bowel disease [IBD]. All consecutive patients undergoing laparoscopic IPAA for IBD in four European expert centres were included and divided into Groups A [SBO during follow-up] and B [no SBO]. From 2005 to 2015, SBO occurred in 41/521 patients [Group A; 8%]. Two-stage IPAA was more frequently complicated by SBO than 3- and modified 2-stage IPAA [12% vs 7% and 4%, p = 0.04]. After multivariate analysis, postoperative morbidity (odds ratio [OR] = 3, 95% confidence interval [CI] = 1.5-7, p = 0.002), stoma-related complications [OR = 3, 95% CI = 1-6, p = 0.03] and long-term incisional hernia [OR = 6, 95% CI = 2-18, p = 0.003] were predictive factors for SBO, while subtotal colectomy as first surgery was an independent protective factor [OR = 0.4, 95% CI = 0.2-0.8, p = 0.002]. In the subgroup of patients receiving restorative proctocolectomy as first operation, stoma-related or other surgical complications and long-term incisional hernia were predictive of SBO. In the patient subgroup of subtotal colectomy as first operation, postoperative morbidity and long-term incisional hernia were predictive of SBO, whereas ulcerative colitis and a laparoscopic approach during the second surgical stage were protective factors. We found that SBO occurred in less than 10% of patients after laparoscopic IPAA. The study also suggested that modified 2-stage IPAA could potentially be safer than procedures with temporary ileostomy [2- and 3-stage IPAA] in terms of SBO occurrence.


Symmetry of bilateral lesions in geographic atrophy in patients with age-related macular degeneration.
As a cause for severe visual loss, geographic atrophy of the retinal pigment epithelium is about half as common as choroidal neovascularization in patients with advanced age-related macular degeneration. To assess symmetry, we determined intraindividual variations of various features of bilateral geographic atrophy in patients with atrophic age-related macular degeneration in a cross-sectional study. Patients were examined with the use of a confocal scanning laser ophthalmoscope (Heidelberg Retina Angiograph; Heidelberg Engineering, Heidelberg, Germany). Digital infrared reflection images (excitation, 830 nm) and fundus autofluorescence images (excitation, 488 nm) were recorded. The eyes of each patient were compared regarding number, size, and convex hull of the atrophic areas with the use of image analysis software and with respect to fundus autofluorescence changes in the junctional zone. Seventy-two patients (mean +/- SD age, 76.3 +/- 7.9 years) were examined. The number of atrophic areas ranged from 1 to 23 (mean +/- SD, 4.9 +/- 4.6); the size of geographic atrophy, from 0.18 to 30.20 (mean +/- SD, 7.0 +/- 6.6) mm(2); and the size of the convex hull, from 0.18 to 39.20 (mean +/- SD, 11.7 +/- 8.4) mm(2). No statistically significant difference was found when comparing these variables between each left and right eye: number, P =.62; size, P =.81; and convex hull, P =.78. Identical patterns of fundus autofluorescence were observed in 43 (80%) of 54 patients. There is intraindividual symmetry in eyes with bilateral geographic atrophy in the presence of a wide range of interindividual variability. The findings are in accordance with the view that age-related macular degeneration is not merely the result of a nonspecific aging process. Symmetric manifestations, rather, reflect specific individual determinants in the pathogenesis and manifestation of the disease.


Flexible, stretchable, and single-molecule-sensitive SERS-active sensor for wearable biosensing applications.
The development of wearable sensors for remote patient monitoring and personalized medicine has led to a revolution in biomedical technology. Plasmonic metasurfaces that enhance Raman scattering signals have recently gained attention as wearable sensors. However, finding a flexible, sensitive, and easy-to-fabricate metasurface has been a challenge for decades. In this paper, a novel wearable device, the flexible, stretchable, and single-molecule-sensetive SERS-active sensor, is proposed. This device offers an unprecedented SERS enhancement factor in the order of 1011, along with other long-desired characteristics for SERS applications such as a high scattering to absorption ratio (∼2.5) and a large hotspot volume (40 nm × 40 nm × 5 nm). To achieve flexibility, we use polydimethylsiloxane (PDMS) as the substrate, which is stable, transparent, and biologically compatible. Our numerical calculations show that the proposed sensor offers reliable SERS performance even under bending (up to 100° angles) or stretching (up to 50% stretch). The easy-to-fabricate and flexible nature of our sensor offers a promising avenue for developing highly sensitive wearable sensors for a range of applications, particularly in the field of personalized medicine and remote patient monitoring.


Oxidative metabolism of clarithromycin in the presence of human liver microsomes. Major role for the cytochrome P4503A (CYP3A) subfamily.
In vitro studies were conducted to identify the hepatic cytochrome P450 (CYP) protein(s) involved in the oxidative metabolism of [14C]clarithromycin (CLAR) in the presence of native human liver microsomes. The identity of the two major CLAR metabolites present in microsome incubates, 14-(R)-hydroxy-CLAR and N-desmethyl-CLAR, was confirmed by MS. Over the CLAR concentration range of 1.0-140 microM, the rate of CLAR 14-(R)-hydroxylation (KM = 48 +/- 17.7 microM; Vmax = 206 +/- 76 pmol/min/mg protein; Vmax/KM = 4.2 +/- 0.21 microliters/min/mg; mean +/- SD, N = 3 livers) and N-demethylation (KM = 59.1 +/- 24.0 microM; Vmax = 189 +/- 52.0 pmol/min/mg protein; Vmax/KM = 3.3 +/- 0.53 microliters/min/mg) conformed to monophasic (saturable) Michaelis-Menten kinetics and was highly correlated (r = 0.90-0.92; p < 0.001; N = 11) with CYP3A-selective erythromycin N-demethylase activity. Ketoconazole (< or = 2.0 microM) or troleadomycin, CYP3A-selective inhibitors, markedly decreased (> or = 99%) the formation of both metabolites, whereas inhibitors selective of other CYP forms were relatively ineffective (< or = 10% inhibition). In agreement with chemical inhibitor studies, CLAR metabolism was only detectable with human B-lymphoblastoid microsomes containing cDNA-expressed CYP3A4 (vs. CYP2C19, CYP2C9, CYP2D6, CYP1A2, CYP2E1, or CYP2A6). Furthermore, the apparent KM characterizing the 14-(R)-hydroxylation and N-demethylation of CLAR in the presence of insect cell microsomes containing cDNA-expressed CYP3A4 (KM = 18-63 microM) was similar to that obtained with native human liver microsomes. Based on the results of this study, it is concluded that the 14-(R)-hydroxylation and N-demethylation of CLAR is primarily mediated by one or more members of the human liver CYP3A subfamily.


Celiac disease screening at a pediatric outpatient clinic: a feasibility study.
Celiac disease (CD) is a common yet largely underdiagnosed disease. This study aimed to test the feasibility of incorporating a non-targeted CD screening in a pediatric outpatient setting and evaluate its short-term impact on children with serological evidence of disease. Over five months, 500 children (aged 2-17 years) attending a general pediatric outpatient clinic in Gothenburg, Sweden, were enrolled and surveyed for current symptoms, quality of life, and background characteristics; 481 children were screened for tissue-transglutaminase antibodies (tTGA); repeated tTGA-positivity was defined as CD autoimmunity (CDA). Children with CDA were investigated for CD and for one year monitored for changes in symptoms, and quality of life. Eleven of 481 (2.3%) screened children had CDA. Children with CDA were younger (median 3.8 years) than those without CDA (8.8 years). No other major between-group differences were reported in background characteristics, symptoms, or quality of life. The screening was well-accepted by the families/participants. During 1-year follow-up, 8 of 11 children with CDA were diagnosed with CD. Children with screening-detected CD reported no significant changes in symptoms and quality of life and the dietary adherence rate was good. Non-targeted screening for CD was feasible in a general pediatric outpatient setting. While hampered by small sample size, our results are in line with previous screening studies indicating that symptoms do not differentiate CDA from non-CDA children. Also, among an overall minimal-symptomatic group of children, diagnosing CD and installation of treatment did not significantly change their well-being during 1-year follow-up.


Estramustine binding protein in human benign prostatic hypertrophy.
Using a highly sensitive radioimmunoassay (RIA) of estramustine binding protein (EMBP) with the antibody raised against rat-EMBP, the concentration of EMBP in human benign prostatic hypertrophy (BPH) tissue was evaluated. Using mechanical separation procedure, EMBP was found significantly higher in the epithelium (10.7 ng/mg protein) than in the stroma (2.9 ng/mg protein). When human BPH was classified into glandular, fibromuscular and mixed type, the concentration of EMBP was the highest in glandular type (15.7 ng/mg protein) and the lowest in fibromuscular type (10.8 ng/mg protein).


Implementation of charged microdroplet-based derivatization of bile acids on a cyclic ion mobility spectrometry-mass spectrometry platform.
Herein, we report the first implementation of charged microdroplet-based derivatization on a commercially-available cyclic ion mobility spectrometry-mass spectrometry platform. We have demonstrated the potential of our approach to improve separability of challenging isomers, but more importantly to rapidly screen derivatization reactions through droplet chemistry. Additionally, the use of cyclic ion mobility separations and tandem mass spectrometry reveals insights into product formation that would be lost with single stage mass spectrometry. Overall, we anticipate broad utility of our methodology owing to the simple design and setup for performing these droplet-based reactions and future work coupling these reactions online with liquid chromatography.


A simplified, sensitive phagocytic assay for malaria cultures facilitated by flow cytometry of differentially-stained cell populations.
Phagocytosis of infected and uninfected erythrocytes is an important feature of malaria infections. Flow cytometry is a useful tool for studying phagocytic uptake of malaria-infected erythrocytes in vitro. However, current approaches are limited by the inability to discriminate between infected and uninfected erythrocytes and a failure to stain the early developmental ring stages of infected erythrocytes. The majority of infected erythrocytes in circulation are of the ring stage and these are therefore important targets to study. In vitro P. falciparum cultures comprising infected and uninfected erythrocytes were labeled and exposed to cells derived from the human monocytic THP-1 cell line. Phagocytosis was assayed by flow cytometry. Dual labeling of Plasmodium DNA and erythrocyte cytoplasm with dihydroethidium and CellTrace™ Violet respectively allowed, for the first time, the detection and enumeration of phagocytes with ingested erythrocytes from both early ring- and late schizont-stage P, falciparum cultures. The sensitivity of the method was tested using varying conditions including phagocyte type (monocytes versus macrophages), parasite stage (rings versus schizonts), and negative (incubation with cytochalasin D) and positive (incubation with immune sera) effectors of phagocytosis. The current assay clearly demonstrated uptake of infected and uninfected erythrocytes exposed to phagocytes; the extent of which was dependent on the conditions mentioned. We describe a simple, sensitive and rapid method for quantifying phagocytosis of P. falciparum-infected erythrocytes, by flow cytometry. This approach can be applied for studying parasite-phagocyte interactions under a variety of conditions. The investigation of phagocytosis of P. falciparum-infected erythrocytes can extend from looking solely at late-staged infected erythrocytes to include early-staged ones as well. It does away with the need to purify infected cells, allowing the study of effects on neighboring uninfected cells. This method may also be translated for use with different types of phagocytes.


Simulated localized wear of resin luting cements for universal adhesive systems with different curing mode.
This study evaluated the simulated localized wear of resin luting cements for universal adhesive systems using different curing modes. Five resin luting cements for universal adhesive systems were evaluated and subsequently subjected to wear challenge in a Leinfelder-Suzuki wear simulation device. Overall, 20 specimens from each resin luting cement were photo-cured for 40 s (dual-cure group), and 20 specimens of each material were not photo-cured (chemical-cure group). Simulated localized wear was generated using a stainless steel ball-bearing antagonist in water slurry of polymethylmethacrylate beads. In addition, scanning electron microscopy (SEM) observations of resin luting cements and wear facets were conducted. Significant differences in simulated wear and SEM observations of wear facets were evident among the materials in the dual- and chemical-cure groups. The simulated wear and SEM observations of wear facets of G-CEM LinkForce and Panavia V5 were not influenced by the curing mode. SEM observations of resin luting cements were material dependent. In most cases, dual curing appears to ensure greater wear resistance of resin luting cements than chemical curing alone. The wear resistance of some resin luting cements appears to be material dependent and is not influenced by the curing mode.


Sum frequency vibrational spectroscopy of leucine molecules adsorbed at air-water interface.
Sum frequency vibrational spectroscopy was used to study adsorption of leucine molecules at air-water interface from solutions with different concentrations and pH values. The surface density and the orientation of the isopropyl head group of the adsorbed leucine molecules could be deduced from the measurements. It was found that the orientation depends on the surface density, but only weakly on bulk pH value at the saturated surface density. The vibrational spectra of the interfacial water molecules appeared to be strongly affected by the charge state of the adsorbed leucine molecules. Enhancement and inversion of polar orientation of interfacial water molecules by surface charges or field controllable by the bulk pH value were observed.


Personalized precision medicine in extreme preterm infants with transient neonatal diabetes mellitus.
Although hyperglycemia is common in neonates, especially preterm infants, a diagnosis of neonatal diabetes mellitus (NDM) is rarely made. NDM can be permanent (45%), transient (45%) or syndromic (10%). Of the 95% of identifiable mutations for NDM, methylation defects in 6q24, KCNJ11, ABCC8, and INS account for the majority. Two cases of transient NDM in extremely preterm, 24 weeks' gestational age (GA) triplets, due to a missense mutation c.685G>A in the KCNJ11 gene are presented. Both patients were successfully transitioned from insulin to Glyburide (Glibenclamide) at 2 months of age. Comprehensive genetic testing with targeted next-generation sequencing and 6q24 methylation analysis helps identify monogenic diabetes early, thereby improving metabolic and glycemic control when patients with potassium channel mutations are started on sulfonylurea (SU) treatment.


Laryngoscopes for difficult airway scenarios: a comparison of the available devices.
According to the American Society of Anesthesiologists, the incidence of difficult intubation in the operating room is 1.2-3.8%; however, in emergency conditions, this rate is higher and reaches even 5.3%. Successful emergency airway management is an essential component of the modern practice of medicine. The aim of the study is to review the literature regarding different devices used to perform endotracheal intubation (Macintosh, Miller, and McCoy laryngoscopes; ETView, GlideScope, TruView, Airtraq, McGrath MAC, Pentax AWS, Trachlight, Shikani, and Bullard) and discuss their clinical and experimental role in difficult airway management. Owing to the development of medical technology, there are an increasing number of videolaryngoscopes and other devices facilitating endotracheal intubation in difficult airway scenarios, including cardiopulmonary resuscitation, cervical spine injury, or face-to-face intubation. Each of these devices may bring benefits in the form of increasing the intubation effectiveness, as well as shortening the procedure, provided that the person performing intubation is familiar with the use of the device.


Phosphatidic acid induces histamine secretion from rat peritoneal mast cells.
Phosphatidic acid produces a selective, dose-dependent release of histamine from rat peritoneal mast cells. The effect is independent of exogenous calcium and probably reflects mobilization of sequestered stores of the cation. The significance of these results is discussed in terms of the possible changes in membrane lipid metabolism involved in mast cell activation.


Characterization of oligopeptide patterns in large protein sets.
Recent sequencing projects and the growth of sequence data banks enable oligopeptide patterns to be characterized on a genome or kingdom level. Several studies have focused on kingdom or habitat classifications based on the abundance of short peptide patterns. There have also been efforts at local structural prediction based on short sequence motifs. Oligopeptide patterns undoubtedly carry valuable information content. Therefore, it is important to characterize these informational peptide patterns to shed light on possible new applications and the pitfalls implicit in neglecting bias in peptide patterns. We have studied four classes of pentapeptide patterns (designated POP, NEP, ORP and URP) in the kingdoms archaea, bacteria and eukaryotes. POP are highly abundant patterns statistically not expected to exist; NEP are patterns that do not exist but are statistically expected to; ORP are patterns unique to a kingdom; and URP are patterns excluded from a kingdom. We used two data sources: the de facto standard of protein knowledge Swiss-Prot, and a set of 386 completely sequenced genomes. For each class of peptides we looked at the 100 most extreme and found both known and unknown sequence features. Most of the known sequence motifs can be explained on the basis of the protein families from which they originate. We find an inherent bias of certain oligopeptide patterns in naturally occurring proteins that cannot be explained solely on the basis of residue distribution in single proteins, kingdoms or databases. We see three predominant categories of patterns: (i) patterns widespread in a kingdom such as those originating from respiratory chain-associated proteins and translation machinery; (ii) proteins with structurally and/or functionally favored patterns, which have not yet been ascribed this role; (iii) multicopy species-specific retrotransposons, only found in the genome set. These categories will affect the accuracy of sequence pattern algorithms that rely mainly on amino acid residue usage. Methods presented in this paper may be used to discover targets for antibiotics, as we identify numerous examples of kingdom-specific antigens among our peptide classes. The methods may also be useful for detecting coding regions of genes.


Non-local dielectric effects in nanoscience.
The physical properties of charges and excitations in nanoscale materials are influenced both by the dielectric properties of the material itself and the surrounding environment. This non-local dielectric effect was first discussed in the context of molecules in solvents over a century ago. In this perspective, we discuss non-local dielectric effects in zero-dimensional, one-dimensional, and two-dimensional nanoscale systems.


Minimally invasive simple prostatectomy: Robotic-assisted versus laparoscopy. A comparative study.
Robotic-assisted simple prostatectomy (RASP) is a novel surgical procedure for the management of obstructive symptoms caused by enlarged prostate glands. Before the introduction of minimally invasive techniques, the standard approach was the open simple prostatectomy (OSP). The aim of our study was to compare intraoperative and perioperative outcomes of robotic (RASP) and laparoscopic (LSP) simple prostatectomy. We retrospectively analyzed data from patients who underwent minimally invasive simple prostatectomy at the Urological Department of Portogruaro Hospital, Portogruaro, and at the Urological Department of "San Bassiano" Hospital, in Bassano del Grappa, from March 2015 to December 2020. Data collected from medical records included age, body mass index, prostate volume, operative time, preoperative International Prostatic Symptoms Score (IPSS), postoperative IPSS, time with drainage, blood transfusion, intraoperative complications, perioperative complications and length of hospital stay. Robotic-assisted (n = 25) and laparoscopic simple prostatectomy (n = 25) were performed with a transvesical approach. No significant differences were observed regarding baseline characteristics, body mass index, prostate volume and IPSS. Operative time was lower in the laparoscopic group (122 min vs 139 min) (p = 0.024), while hospital stay was lower in the robotic group (4 days vs 6 days) (p = 0.047). Robotic-assisted simple prostatectomy is a safe technique with results comparable to laparoscopic simple prostatectomy, encompassing the advantage of a shorter hospitalization. Considering the costs and the limited availability of robotic-assisted simple prostatectomy, laparoscopic simple prostatectomy is a valid and safe alternative for experienced surgeons.


Changes in chromosomal proteins in colon cancer: the complexity and DNA-binding properties of tumor-associated proteins and evidence for their association with the malignant state in human colonic epithelium.
The properties of two classes of nonhistone nuclear proteins (NHNP) whose appearance has been correlated with the process of carcinogenesis in the colonic epithelium of rats given 1,2-dimethylhydrazine (DMH) have been investigated. NHNP isolated from adenocarcinomas of the colon were tested for their binding to homologous DNA. The tumor-specific protein class TNP1 (molecular weight ca. 44,000) shows high affinity for DNA while the second class of tumor-specific proteins (TNP2; molecular weight ca. 62,000) does not bind to DNA. When tumor chromatin is subjected to a limited digestion with DNAse I under conditions that are known to preferentially digest active genes, the TNP1 proteins are selectively released, suggesting that this protein fraction is associated with the actively transcribing portions of the genome. The complexity of both tumor-specific protein classes has also been analyzed by two-dimensional gel electrophoresis. This method reveals a high degree of complexity both in TNP1 and TNP2. Protein classes similar to TNP1 and TNP2 that are also found in human colonic adenocarcinomas are not detectable in polyps from familial polyposis-affected patients at times when no sign of malignancy has yet appeared.


Haematological changes in adults receiving a zidovudine-containing HAART regimen in combination with cotrimoxazole in Côte d'Ivoire.
Neutropenia is the most frequent side effect of cotrimoxazole in sub-Saharan Africa. We estimated the incidence of haematological disorders during the first 6 months of a zidovudine-containing highly active antiretroviral therapy (HAART) regimen in sub-Saharan African adults receiving cotrimoxazole. Prospective cohort study in Abidjan, with blood cell count measurement at baseline (HAART initiation), month 1, month 3 and month 6. A total of 498 adults [baseline: 80% currently on cotrimoxazole prophylaxis; median CD4 count 237/mm3 [interquartile range (IQR) 181;316]; median neutrophil count 1647/mm3 (IQR 1221;2256); median haemoglobin 113 g/l (IQR 102;122)] started zidovudine (AZT)/lamivudine/efavirenz. During follow-up, 118 patients had a grade 3-4 neutropenia [(56.3/100 person-years (PY)], 23 had a grade 3-4 anaemia (9.6/100 PY) and no cases of grade 3-4 thrombocytopenia. Of the 118 patients with grade 3-4 neutropenia, 86 (73%) had to stop cotrimoxazole because neutropenia persisted, and one (<1%) had to stop AZT because of persistent neutropenia after cotrimoxazole was stopped (neutropenia-related HAART modification: 0.4/100 PY). Of the 23 patients with grade 3-4 anaemia, 11 had to stop AZT (anaemia-related HAART modification: 4.4/100 PY). In patients who stopped cotrimoxazole but not AZT, the median gain in neutrophils at 1 month was +540/mm3 (IQR +150;+896). At baseline, most patients had a normal neutrophil count and 80% of them were already receiving cotrimoxazole. An unexpectedly high rate of grade 3-4 neutropenia occurred shortly after introduction of AZT. Almost all of the persistent severe neutropenia disappeared after cotrimoxazole was stopped. This suggests an accentuated drug interaction between the two drugs in these sub-Saharan African individuals. Grade 3-4 anaemia was much less frequent, but remained the first cause of AZT discontinuation.


IgG4-related inflammatory abdominal aortic aneurysm associated with autoimmune pancreatitis.
An association between autoimmune pancreatitis (AIP) and inflammatory abdominal aortic aneurysm (AAA) has never been reported. Reported herein is a case of IgG4-related inflammatory AAA accompanying metachronous AIP. A 77-year-old man presented with malaise and intermittent lower abdominal pain. Radiological examination showed inflammatory AAA and right hydronephrosis caused by retroperitoneal fibrosis. Surgical correction of the AAA was performed, but high levels of systemic inflammatory markers persisted. Four months after surgery, the patient presented with epigastric pain, backache, and jaundice. His serum IgG4 concentration was high (571 mg/mL), and he was diagnosed with AIP, based on clinical and radiological findings. Corticosteroid therapy resulted in improvement of the clinical findings and lowered his serum IgG4 levels. Subsequent histological examination of a specimen from the aortic wall showed irregular proliferation of fibroblastic and myofibroblastic cells, severe lymphoplasmacytic infiltration, and obliterative phlebitis in the adventitia. Furthermore, on immunohistochemistry many plasma cells within the lesion were found to be positive for IgG4. These findings suggest that inflammatory AAA has a pathological process similar to that of AIP, and that some cases of inflammatory AAA and retroperitoneal fibrosis may be aortic and periaortic lesions of an IgG4-related sclerosing disease.


Research on the elimination of low-concentration formaldehyde by Ag loaded onto Mn/CeO2 catalyst at room temperature.
Formaldehyde (HCHO) is one of the major air pollutants, and its effective removal at room temperature has proven to be a great challenge. In this study, an Ag/Mn/CeO2 catalyst for the catalytic oxidation of low-concentration HCHO at room temperature was prepared by a hydrothermal-calcination method. The removal performance of the Ag/Mn/CeO2 catalyst for HCHO was systematically studied, and its surface chemical properties and microstructure were analyzed. The incorporation of Ag did not change the mesoporous structure of the Mn/CeO2 catalyst but reduced the pore size and specific surface area. The Ag species included metallic Ag as the main component and part of Ag+. The well-dispersed Ag species on the catalyst provided sufficient active sites for the catalytic oxidation of HCHO. The more the Ag active sites, the more the lattice defects and oxygen vacancies generated from the interaction of Ag with Mn/CeO2. Precisely because of this, the Ag/Mn/CeO2 catalyst exhibited high catalytic activity for HCHO at room temperature with a removal efficiency of 96.76% within 22 h, which is 22.91% higher than that of the Mn/CeO2 catalyst. Moreover, the Ag/Mn/CeO2 catalyst showed good cycling stability and the removal efficiency reached 85.77% after five cycles. Therefore, the as-prepared catalyst is an effective and sustainable material that can be used to remove HCHO from actual indoor polluted air. This paper provides ideas for the research and development of efficient catalysts.


Efficacy of perampanel: a review of pooled data.
Perampanel (PER) has been tested in three randomized placebo-controlled prospective phase III trials as an adjunctive antiepileptic drug (AED) in adult and adolescent patients age 12 years and older who had ongoing focal epileptic seizures despite receiving one to three AEDs. Patients were randomized to once-daily placebo or maintenance dosages of 2, 4, or 8 mg of PER daily in one trial or to dosages of 8 and 12 mg of PER in the other two studies. Baseline and double-blind titration periods comprised 6 weeks each before a 12-week maintenance phase. Primary endpoints were median change in partial seizure frequency (baseline vs. double-blind phase), and the percentage of patients achieving >50% reduction in seizure frequency (so-called responders, baseline vs. maintenance). All patients had the opportunity to enter an open follow-up study that allowed a titration of PER of up to 12 mg. Pooled data of all three studies included 1,478 patients. There was a statistically significant median change in seizure frequency and responder rates with PER dosages of 4, 8, and 12 mg (p < 0.01, each dose vs. placebo). Both the results of each trial and the pooled data are reported in this review. For the open follow-up study, 1,218 patients were recruited. Ninety-one percent finally reached maintenance dosages of 10 or 12 mg. After 1 year, both median seizure reduction and responder rates reached almost 50%.


Tandem Aldol Condensation - Platinacycle-Catalyzed Addition Reactions of Aldehydes, Methyl Ketones and Arylboronic Acids.
Tandem aldol condensation of aldehydes with methyl ketones followed by anionic four-electron donor-based (Type I) platinacycle-catalyzed addition reactions of arylboronic acids to form β-arylated ketones is described. Good to excellent yields of β-arylated ketones were obtained for the tandem reactions of aromatic/aliphatic aldehydes, methyl ketones and arylboronic acids, and moderate yields were observed for the tandem reaction with α, β-unsaturated aldehydes as the aldehyde source.


The Utility of Metagenomic Next-Generation Sequencing (mNGS) in the Management of Patients With Bronchiectasis: A Single-Center Retrospective Study of 93 Cases.
Bronchiectasis is a chronic inflammatory respiratory disease mainly caused by pathogenic infections. However, standard methods of pathogen detection show prolonged cycle durations and unsatisfactory sensitivity and detection rates. Macrogenomic next-generation sequencing (mNGS) emerges as a promising technique for swift, effective, and unbiased pathogen detection and subsequent data interpretation. Here, a retrospective analysis of 93 patients with suspected bronchiectasis was performed to assess the clinical applicability of mNGS. Bronchoalveolar alveolar lavage fluid (BALF) samples were collected from these subjects, followed by standard assays and mNGS separately. The turnaround time, detection rate, and pathogen identification using mNGS were compared with those of standard methods. mNGS identified a greater number of bacteria (72 vs 16), fungi (26 vs 19), and viruses (14 vs 0) than standard methods. Specifically, the commonly identified bacteria were Haemophilus, Mycobacterium intracellulare, Pseudomonas, and Streptococcus pneumoniae, while the most detected fungi were Aspergillus and the most prevalent viruses were human herpesviruses. Of note, 29 out of 30 patients (96.67%) who received optimized treatment strategies based on mNGS results experienced recovery. Collectively, these findings suggest that mNGS has the potential to improve the diagnosis and treatment of bronchiectasis patients by enabling rapid and precise pathogen detection, which can lead to timely and effective treatment strategies.


[Use of the method of enriching of soil samples with calcium carbonate for isolation of Actinomyces].
Possible application of a modified procedure for treatment of soil samples with calcium carbonate under humid conditions to isolation of actinomycetes was studied. The procedure proved to be rather efficient in regard to increasing the number of the isolates including representatives of rare genera as compared with the routine methods. The number of microorganisms isolated from the soil samples treated with calcium carbonate under humid conditions was 2 orders higher than that from untreated soil samples. 1033 actinomycete cultures were isolated from the soil samples treated in accordance with the above procedure and only 597 cultures were isolated from the untreated soil samples. The antibiotic activity of the isolates was studied and their taxonomic position at the genus level was determined. The preliminary treatment of soil samples equally stimulated development of actinomycetes belonging to different genera. It is advisable that the described procedure for isolation of actinomycetes from soil samples in screening of cultures producing new antibiotics be used in combination with other selective methods of isolation.


Protocol for evaluation of automated blood cell counters. International Committee for Standardization in Haematology (ICSH).
This protocol is proposed for the evaluation of automated blood cell counters to assess the performance, advantages and limitations of such instruments. It is based on the International Committee for Standardization in Haematology (ICSH) 'Protocol for type testing equipment and apparatus used for haematological analysis' (1978a) and the British Committee for Standardization in Haematology 'Guidelines for the evaluation of instruments used in haematology' (Shinton, England & Kennedy, 1982). The document has been prepared by the ICSH Panel on Cytometry after discussion with colleagues. This tentative protocol will be reviewed 1 year after publication, in accordance with the ICSH rules, before it is adopted as a definitive standard.


More rapid progression to AIDS in older HIV-infected people: the role of CD4+ T-cell counts.
The tendency for older people with HIV infection to progress more rapidly to AIDS than younger people was studied in a group of 111 anti-HIV-positive haemophiliacs followed for up to 10 years from seroconversion. After 7 years of seropositivity, those aged over 30 years at the time of the first positive anti-HIV test had a cumulative progression rate to AIDS of 50%, compared with only 12% for those aged 10-19 years (Kaplan-Meier estimates). Overall, the relative risk of developing AIDS by any given time after seroconversion was 1.45 for each 10 year increase in age (p = 0.002; 95% confidence limits of 1.15, 1.85; Cox proportional hazards model). After adjustment for the CD4+ T-cell count (median of 10 count measurements per patient, fitted as a time-dependent covariate), the relative risk fell to 1.31 but remained statistically significant (p less than 0.05; 95% confidence limits of 1.03, 1.67). This implies that older people may be at higher risk of progression than their younger counterparts, even if their CD4+ T-cell counts are the same. Hence, prophylaxis against opportunistic infections may be indicated at higher CD4+ T-cell counts in older people than in younger people.


French invasive Asian tiger mosquito populations harbor reduced bacterial microbiota and genetic diversity compared to Vietnamese autochthonous relatives.
The Asian tiger mosquito Aedes albopictus is one of the most significant pathogen vectors of the twenty-first century. Originating from Asia, it has invaded a wide range of eco-climatic regions worldwide. The insect-associated microbiota is now recognized to play a significant role in host biology. While genetic diversity bottlenecks are known to result from biological invasions, the resulting shifts in host-associated microbiota diversity has not been thoroughly investigated. To address this subject, we compared four autochthonous Ae. albopictus populations in Vietnam, the native area of Ae. albopictus, and three populations recently introduced to Metropolitan France, with the aim of documenting whether these populations display differences in host genotype and bacterial microbiota. Population-level genetic diversity (microsatellite markers and COI haplotype) and bacterial diversity (16S rDNA metabarcoding) were compared between field-caught mosquitoes. Bacterial microbiota from the whole insect bodies were largely dominated by Wolbachia pipientis. Targeted analysis of the gut microbiota revealed a greater bacterial diversity in which a fraction was common between French and Vietnamese populations. The genus Dysgonomonas was the most prevalent and abundant across all studied populations. Overall genetic diversities of both hosts and bacterial microbiota were significantly reduced in recently established populations of France compared to the autochthonous populations of Vietnam. These results open up many important avenues of investigation in order to link the process of geographical invasion to shifts in commensal and symbiotic microbiome communities, as such shifts may have dramatic impacts on the biology and/or vector competence of invading hematophagous insects.


Stem cells: potency, plasticity and public perception.
Production of chimaeras with embryonal carcinoma and embryonic stem cells enabled a very thorough investigation of the potency of these cells in the mouse. Human embryonal carcinoma and embryonic stem cell differ from their murine counterparts in a number of respects and, for obvious reasons, their potency is more difficult to assess. Recently, findings attesting to a surprising degree of plasticity of cells from adults have begun to emerge, which, aside from offering a possible further route to stem cell therapy, raise intriguing questions about the importance of lineage in the process of cellular diversification. Biomedical research is widely perceived to be advancing too fast to allow proper consideration of the implications of its clinical applications. Whilst this was clearly not true in the case of human in vitro fertilization, it has some validity regarding stem cell therapy, even though many of the issues are common to both. Casual use of the term 'embryo' proved unhelpful in the past debate on whether research on early stages of human development should be permitted. Likewise, introduction of the term 'therapeutic cloning' has complicated the present one regarding extension of such research to stem cell therapy.


Ionic liquids as precursors for efficient mesoporous iron-nitrogen-doped oxygen reduction electrocatalysts.
A ferrocene-based ionic liquid (Fe-IL) is used as a metal-containing feedstock with a nitrogen-enriched ionic liquid (N-IL) as a compatible nitrogen content modulator to prepare a novel type of non-precious-metal-nitrogen-carbon (M-N-C) catalysts, which feature ordered mesoporous structure consisting of uniform iron oxide nanoparticles embedded into N-enriched carbons. The catalyst Fe(10) @NOMC exhibits comparable catalytic activity but superior long-term stability to 20 wt % Pt/C for ORR with four-electron transfer pathway under alkaline conditions. Such outstanding catalytic performance is ascribed to the populated Fe (Fe3 O4 ) and N (N2) active sites with synergetic chemical coupling as well as the ordered mesoporous structure and high surface area endowed by both the versatile precursors and the synthetic strategy, which also open new avenues for the development of M-N-C catalytic materials.


Compartmental modeling of 11C-HOMADAM binding to the serotonin transporter in the healthy human brain.
The novel PET radioligand (11)C-N,N-dimethyl-2-(2'-amino-4'-hydroxymethylphenylthio)benzylamine ((11)C-HOMADAM) binds with high affinity and selectively to the serotonin transporter (SERT). The purpose of this study was to develop a reliable kinetic model to describe the uptake of (11)C-HOMADAM in the healthy human brain. Eight volunteers participated in the study; 5 of them were fitted with arterial catheters for blood sampling and all were scanned on a high-resolution research tomograph after the injection of (11)C-HOMADAM. Regional distribution volumes and binding potentials were calculated with 2- and 4-parameter arterial-input compartment models, a 3-parameter reference tissue compartment model, and the Logan graphic approach. The 2-parameter arterial-input compartment model was statistically superior to the 4-parameter model and described all brain regions. Calculated binding potentials agreed well between the arterial-input model and the reference tissue model when the cerebellum was used as the reference tissue. The Logan graphic approach was not able to estimate the higher concentration of SERT in the dorsal raphe than in the midbrain. (11)C-HOMADAM is a highly promising radioligand with high ratios of specific binding to nonspecific binding in known SERT-rich structures, such as the raphe nuclei. The 3-parameter reference tissue model approach permits a simplified quantitatively accurate method for estimating SERT binding potentials.


Protein Arginine Methyltransferase 6 (Prmt6) Is Essential for Early Zebrafish Development through the Direct Suppression of gadd45αa Stress Sensor Gene.
Histone lysine methylation is important in early zebrafish development; however, the role of histone arginine methylation in this process remains unclear. H3R2me2a, generated by protein arginine methyltransferase 6 (Prmt6), is a repressive mark. To explore the role of Prmt6 and H3R2me2a during zebrafish embryogenesis, we identified the maternal characteristic of prmt6 and designed two prmt6-specific morpholino-oligos (MOs) to study its importance in early development, application of which led to early epiboly defects and significantly reduced the level of H3R2me2a marks. prmt6 mRNA could rescue the epiboly defects and the H3R2me2a reduction in the prmt6 morphants. Functionally, microarray data demonstrated that growth arrest and DNA damage-inducible, α, a (gadd45αa) was a significantly up-regulated gene in MO-treated embryos, the activity of which was linked to the activation of the p38/JNK pathway and apoptosis. Importantly, gadd45αa MO and p38/JNK inhibitors could partially rescue the defect of prmt6 morphants, the downstream targets of Prmt6, and the apoptosis ratios of the prmt6 morphants. Moreover, the results of ChIP quantitative real time PCR and luciferase reporter assay indicated that gadd45αa is a repressive target of Prmt6. Taken together, these results suggest that maternal Prmt6 is essential to early zebrafish development by directly repressing gadd45αa.


Does cosmic-ray-induced heterogeneous chemistry influence stratospheric polar ozone loss?
Cosmic-ray (CR) -induced heterogeneous reactions of halogenated species have been suggested to play the dominant role in causing the Antarctic ozone hole. However, measurements of total ozone in Antarctica do not show a compact and significant correlation with CR activity. Further, a substantial CR-induced heterogeneous loss of chlorofluorocarbons is incompatible with multiyear satellite observations of N2O and CFC-12. Thus, CR-induced heterogeneous reactions cannot be considered as an alternative mechanism causing the Antarctic ozone hole.