Effect of antimicrotubule agents on secretion of relaxin by large luteal cells derived from pregnant swine.
The role of microtubules in the modulation of secretion of the ovarian protein hormone, relaxin, by porcine large luteal cells (LLCs) was examined by use of a reverse hemolytic plaque assay. In this assay, luteal cells were cocultured in monolayers with protein-A-coupled ovine erythrocytes. In the presence of porcine relaxin antiserum and complement, a zone of hemolysis, a plaque, developed around relaxin-releasing LLCs. The rate of development of plaques in time-course experiments (1-12 h) and the detection of discontinuities in the pattern of plaque formation were used in this study as an index of the rate of relaxin release and a method to detect differential responsiveness of individual LLCs, respectively. Monolayers were bathed in medium containing three different antimicrotubule agents that bind to tubulin and induce microtubule loss through depolymerization. Exposure of luteal cell-containing monolayers to colchicine (100 microM), vinblastine (1 microM), and nocadazole (30 microM) resulted in a reduction in the rate of relaxin-induced plaque formation by LLCs, unequivocal evidence of an inhibitory effect of all three of these antimicrotubule agents of the rate of basal relaxin release. However, this suppressive effect on relaxin release was strictly time related. None of the antimicrotubule agents significantly affected plaque formation until 3 h of incubation. Thereafter, a plateau of plaque formation was observed in antimicrotubule-treated monolayers over approximately 3-8 h of incubation, indicating substantial slowing or cessation of relaxin release by LLCs during this period. During this middle phase of the incubation, plaque formation in antimicrotubule-treated monolayers was significantly lower (P less than 0.05) than that in controls. By 8-12 h of incubation plaque formation was still significantly suppressed in vinblastine-treated monolayers, but not, however, in colchicine- or nocadozole-treated monolayers. Treatment with beta-lumicolchicine (100 microM; a form of colchicine that fails to bind to tubulin) resulted in no significant change in the rate of plaque formation compared with that in controls. We conclude that microtubule-assisted transport forms an important intracellular mechanism that assists basal release of an ovarian peptide hormone, relaxin. Moreover, the discontinuity of plaque formation in the middle phase of the incubation is consistent with the view that a subpopulation of LLCs responds preferentially to antimicrotubule agents. However, the exact nature and physiological role(s) of microtubules in the control of relaxin secretion and the interrelationships of microtubules with other intracellular regulatory events remain to be clearly defined.


Design and analysis of food frequency questionnaires--review and novel method.
A novel method is proposed for the design and analysis of a food frequency questionnaire facilitated by a computer program. The method relies on a weighed inventory undertaken by a sample of the population that is combined into a database along with the nutrient analyses of the foods consumed. The program provides an interface that enables the researcher to see the total amount of each food that was consumed, decide on what questions to ask and allocate the relevant foods to these questions. The approach means that all foods consumed can appear in the questionnaire. The nutrient analysis for each question is a weighted mean depending on the inventory. The portion weight employed for each question is derived from the weights of foods consumed in the inventory and any misperceptions in the frequency of consumption of all the subjects are corrected by calculating a factor to multiply by the frequency from the questionnaire. A viability study established the practicability of the method.


Modification of diethylnitrosamine liver carcinogenesis with phenobarbital but not with immunosuppression.
Administration of 40 ppm diethylnitrosamine (DENA) in the drinking water for 10 weeks to male Fischer rats led to hepatocellular carcinoma in 100 percent with metastasis to the lung in 40 percent, of the animals living for the full experimental period of 20 weeks. Concurrent feeding of phenobarbital and DENA for 10 weeks produced cancer of the liver in 77 percent of the animals, but only 9 percent had metastases in the lung. A brief regimen of DENA for 4 weeks, followed by 16 weeks of observation, induced cancer of the liver in only 13 percent of the rats. Administration of phenobarbital, begun 1 week after cessation of DENA intake and terminated at week 20, led to liver cancer in 64 percent of the rodents. Hydroxyurea had no effect on this enhancement. Treatment with a purified gamma fraction of antilymphocytic serum after the DENA did not influence the outcome. Thus phenobarbital given together with DENA reduced the severity of the carcinogenic process, but when it was given after the hepatocarcinogen, it increased the effect.


Effects of adjunctive treatment with combined cytokines in a neutropenic mouse model of multidrug-resistant Enterococcus faecalis septicemia.
To examine whether the antienterococcal efficacy of a regimen of gentamicin plus vancomycin combined with granulocyte colony-stimulating factor (G-CSF) is enhanced by concurrent therapy with interferon-gamma (IFN-gamma). Hospital laboratory. Mice rendered neutropenic by cyclophosphamide were intraperitoneally inoculated with a gentamicin- and vancomycin-resistant Enterococcus faecalis isolate. Infected animals were randomized into treatment groups that received G-CSF alone or in combination with various dosages of IFN-gamma. Additional groups of animals received vancomycin; G-CSF, G-CSF plus vancomycin, IFN-gamma, and G-CSF; or vancomycin with both cytokines. Addition of IFN-gamma to G-CSF regimen resulted in no significant change (p>0.05) in survival, compared with treatment with G-CSF alone. Also, the antienterococcal efficacy of antibiotic plus G-CSF was not modified by coadministration of IFN-gamma. This study suggests that adjunctive application of combined cytokines may not be more beneficial than only G-CSF in combination with an antibiotic to treat multidrug-resistant enterococcal infection.


[Long-term outcome of initial treatment with Bacillus Calmette-Guérin for carcinoma in situ of the upper urinary tract].
To assess appropriate treatment strategies for transitional cell carcinoma in situ (CIS) of the upper urinary tract (UUT), we evaluated the long-term outcome of Bacillus Calmette-Guérin (BCG) perfusion therapy for CIS of UUT. We retrospectively reviewed the medical records of 24 patients who underwent BCG perfusion therapy for CIS of UUT between August 1993 and August 2009. Patients received at least one course of BCG (once weekly for 6 weeks). The median follow-up period was 48.5 months (range 16-201 months). In 23 patients (96%), cytology became negative after one course of BCG perfusion and 12 patients (50%) remained disease-free for a median follow-up of 38. 5 months. In 11 patients positive cytology recurred, and in five of them nephroureterectomy was performed after radiologic studies showed the presence of a tumor in the UUT. Histopathology showed invasive tumor (pT3) in all cases, and three of them experienced distant metastases after surgery. In conclusion, BCG perfusion therapy is effective for the treatment of CIS of UUT with long-term follow-up. However, in cases with a poor response or recurrence of CIS, there is a high risk of developing invasive tumor. Surgical intervention should be immediately considered in such cases after the first course of BCG perfusion therapy.


[Teaching in orthopaedic and trauma surgery].
The scramble for available places to study medicine in Germany continues. One of the core points within the "Master Plan 2020", which has been recently adopted, is the reform of teaching concepts at German medical faculties. In the future, students of human medicine are to be trained primarily based upon competence-oriented, integrated curricula. The present paper is intended to present modern teaching concepts and methods, as well as current examination formats, which play a special role in student training in orthopaedics and traumatology. Knowledge in the field of medical and evidence-based treatment strategies, clinical examination techniques, diagnostic/therapeutic procedures and hygienic working conditions are especially important. Individual courses with a focus on research-, practice- and competence-orientation are intended to help the students grasp the depth of the subject.


Synthesis and enantioselectivity of the antiviral effects of (R,Z)-,(S,Z)-methylenecyclopropane analogues of purine nucleosides and phosphoralaninate prodrugs: influence of heterocyclic base, type of virus and host cells.
A series of R and S enantiomers of 2-aminopurine methylenecyclopropane analogues of nucleosides was synthesized. Two diastereoisomeric lipophilic phosphate prodrugs derived from R and S enantiomers of 2,6-diaminopurine analogue were also prepared. Enantioselectivity (diastereoselectivity in case of prodrugs) of in vitro antiviral effects was investigated with human and murine cytomegalovirus (HCMV and MCMV, respectively), herpes simplex virus types 1 and 2 (HSV-1 and HSV-2, respectively), human immunodeficiency virus type 1 (HIV-1), hepatitis B virus (HBV), Epstein-Barr virus (EBV) and varicella zoster virus (VZV). Strong differences in enantioselectivity were found between the R and S enantiomers of adenine analogue and enantiomeric 2-aminopurine analogues. Thus, the enantiomers of adenine analogue were equipotent against HCMV but not MCMV, where the S enantiomer is strongly preferred. The same S preference was found throughout the 2-aminopurine series for both HCMV and MCMV. In contrast, R-synadenol in HIV-1 assays was the best agent, whereas the S enantiomers of moderately effective 2-amino-6-cyclopropylamino and 2-amino-6-methoxypurine analogues were preferred. Little enantiomeric preference was found for R and S enantiomers of synadenol and the corresponding enantiomers of 2,6-diaminopurine analogue against HBV. A mixed pattern of enantioselectivity was observed for EBV depending on the type of host cells and assay. Against VZV, the R and S enantiomers of adenine analogue were equipotent or almost equipotent, but throughout the series of 2-aminopurine analogues a distinct preference for the S enantiomers was found. The stereoselectivity pattern of both diastereoisomeric prodrugs mostly followed enantioselectivity of the parent analogues. The varying enantioselectivities in the series of purine methylenecyclopropane analogues are probably a consequence of differences in the mechanisms of action in different virus/host cell systems.


Structure of a liganded type 2 non-specific lipid-transfer protein from wheat and the molecular basis of lipid binding.
In plants, a family of ubiquitous proteins named non-specific lipid-transfer proteins (ns-LTPs) facilitates the transfer of fatty acids, phospholipids and steroids between membranes. Recent data suggest that these secreted proteins play a key role in the formation of cuticular wax layers and in defence mechanisms against pathogens. In this study, X-ray crystallography has been used to examine the structural details of the interaction between a wheat type 2 ns-LTP and a lipid, L-alpha-palmitoyl-phosphatidyl glycerol. This crystal structure was solved ab initio at 1.12 A resolution by direct methods. The typical alpha-helical bundle fold of this protein is maintained by four disulfide bridges and delineates two hydrophobic cavities. The inner surface of the main cavity is lined by non-polar residues that provide a hydrophobic environment for the palmitoyl moiety of the lipid. The head-group region of this lipid protrudes from the surface and makes several polar interactions with a conserved patch of basic residues at the entrance of the pocket. The alkyl chain of a second lipid is bound within an adjacent smaller cavity. The structure shows that binding of the lipid tails to the protein involves extensive hydrophobic interactions.


Microplastics in Baltic bottom sediments: Quantification procedures and first results.
Microplastics in the marine environment are known as a global ecological problem but there are still no standardized analysis procedures for their quantification. The first breakthrough in this direction was the NOAA Laboratory Methods for quantifying synthetic particles in water and sediments, but fibers numbers have been found to be underestimated with this approach. We propose modifications for these methods that will allow us to analyze microplastics in bottom sediments, including small fibers. Addition of an internal standard to sediment samples and occasional empty runs are advised for analysis quality control. The microplastics extraction efficiency using the proposed modifications is 92±7%. Distribution of microplastics in bottom sediments of the Russian part of the Baltic Sea is presented. Microplastic particles were found in all of the samples with an average concentration of 34±10 items/kg DW and have the same order of magnitude as neighbor studies reported.


One species or several? Discordant patterns of geographic variation between allozymes and mtDNA sequences among spiders in the genus Metepeira (Araneae: Araneidae).
Paradoxically, an allozyme study of Metepeira "spinipes" (sensu lato) demonstrated extensive gene flow among four populations whose members are nevertheless morphologically and behaviorally distinct. Initially, the authors tentatively concluded that the populations exhibited panmixis and suggested that local environmental effects accounted for the apparent morphological and behavioral differences. However, they later concluded that such differences were too great to be accounted for by the environment alone and that the four populations actually represented three different species. To confirm that the allozyme results were, in fact, artifactual, we reexamined the relationships among these populations by sequencing a portion of the 12S mtDNA ribosomal subunit. In contrast to the allozyme result, our results demonstrate good agreement between patterns of genetic and morphological/behavioral variation. We suggest (1) that the allozyme allele frequencies are homogenized by balancing selection, not gene flow as was previously concluded, and therefore (2) that this study provides another instance in which inferences about population structures from allozyme data are misleading.


Evaluation of Early Reperfusion Criteria in Acute Ischemic Stroke.
Though still debated, early reperfusion is increasingly used as a biomarker for clinical outcome. However, the lack of a standard definition hinders the assessment of reperfusion therapies and study comparisons. The objective was to determine the optimal early reperfusion criteria that predicts clinical outcome in ischemic stroke. Early reperfusion was assessed voxel-wise in 57 patients within 6 hours of symptom onset. The performance of the time to peak (TTP), the mean transit time (MTT), and the time to maximum of residue function (Tmax ) at various delays thresholds in predicting the neurological response (based on the National Institutes of Health Stroke Scale) and the functional outcome (modified Rankin scale ≤1) at 1 month were compared. A receiver operating characteristics (ROC) analysis determined the optimal extent of reperfusion. A novel unsupervised classification of reperfusion using group-based trajectory modeling (GBTM) was evaluated. MTT had a lower performance than TTP and Tmax in predicting the neurological response (P = .008 vs. TTP and P = .006 vs. Tmax ) or the functional outcome (P = .0006 vs. TTP; P = .002 vs. Tmax ). No delay threshold had a significantly higher predictive value than another. The optimal percentage of reperfusion was dependent on the outcome scale (P < .001). The GBTM-based classification of reperfusion was closely associated with the clinical outcome and had a similar accuracy compared to ROC-based classification. TTP and Tmax should be preferred to MTT in defining early reperfusion. GBTM provided a clinically relevant reperfusion classification that does not require prespecified delay thresholds or clinical outcomes.


Protease activity in extracellular products secreted in vitro by trophozoites of Giardia duodenalis.
There are evidences that Giardia trophozoites contain and/or release proteolytic enzymes that may be implicated in pathogenesis of giardiasis. This report describes a preliminary characterization of the proteolytic activity in excretory/secretory (E/S) products of Giardia duodenalis trophozoites of an axenic Brazilian strain (BTU-11) and the reference strain Portland 1 (P1). The protease activity of E/S products in conditioned medium by trophozoites of each strain was analyzed using substrate (gelatin and collagen) impregnated SDS-PAGE and hemoglobin assay. The protease characterization was based on inhibition assays including synthetic inhibitors. Proteolytic products were detected in the conditioned medium by trophozoites of both assayed strains. In the gels containing copolymerized gelatin and collagen, E/S products promoted degradation of the substrates and the most evident proteolysis zones were distributed in the migration regions of 77 to 18 kDa and 145 to 18 kDa, respectively, in the patterns of gelatinolytic and collagenolytic activities. Degradation of hemoglobin was also observed, and the pattern of hydrolysis was similar in both E/S products assayed. Inhibition assays showed that the main proteolytic activity in both E/S products is due to cysteine proteases although the presence of serine proteases was also indicated, mainly in the hydrolysis of hemoglobin.


Changes in ethnic identity and competence in middle adolescents.
Evidence suggests that the construct of ethnic identity (EI) in adolescents changes over time and is related to factors including social connectedness, social competence, coping, and self-worth. Transitional changes of EI in middle adolescence have not been studied. This analysis, part of a longitudinal study of health behaviors, explored how EI changes from the first year of high school to the last, and it examined relationships among EI, social connectedness, indicators of social competence, coping, and self-worth in 602 youth (49% Hispanic, 59% female). EI was significantly related to social connectedness, social competence, coping, and self-worth as adolescents entered and prepared to exit high school. EI increased significantly over time for both genders and for Hispanics more so than for non-Hispanics. Findings support and extend the conceptual framework and previous research. Socialization with one's ethnic group is associated with developing competence and self-worth. EI development continues throughout high school. Findings extend our understanding of the salience of EI in adolescents in transition throughout high school. Strengthening EI may contribute to adolescents' feelings of self-worth, skills in coping, and competence within an ethnically diverse society. Nurses might plan interventions to strengthen EI throughout adolescence.


The gastrointestinal manifestations of AIDS.
Gastrointestinal disease in AIDS is common and is due to opportunistic infections, aggressive malignancy and possible direct HIV enteropathy. Disabling gastrointestinal symptoms are prominent both in patients with established AIDS and in patients with earlier stages of HIV infection. We report the cases of 160 patients with AIDS who underwent gastroenterological investigations at St Vincent's Hospital, Sydney, between November 1983 to October 1987. Of these, 127 had the diagnosis of AIDS established prior to referral and 33 patients had the diagnosis of AIDS established as a result of gastroenterological investigations. Diarrhoea and weight loss (88%) were the most frequent reasons for undertaking gastroenterological investigations. Swallowing disorders (47%), abdominal pain (20%), oral and perianal disease (74%) and evidence of hepatobiliary disease were the other major indications for investigation. In 90% of cases there was evidence of concurrent and active gastrointestinal disease at two or more sites within the alimentary tract. Results from this series reveal a wide range of infectious pathogens: viral (Cytomegalovirus, Herpes simplex), bacterial (Mycobacterium avium intracellulare) and parasitic (Cryptosporidium, Isospora belli). Kaposi's sarcoma and non-Hodgkin's lymphoma were the only malignancies detected in this series. Gastrointestinal disease associated with HIV infection is common, and contributes significantly to its overall morbidity and mortality. Moreover, chronic diarrhoea, weight loss and malnutrition may also contribute to the overall immunodeficiency.


[Inverted papilloma of the pyelo-ureteral junction, current literature data].
The authors report the fourth case of inverted papilloma of the ureteropelvic junction since the first description of this lesion in 1963 by Potts and Hirst. In view of the rarity of this lesion and its unusual presentation, they conducted an exhaustive review of the literature concerning inverted papillomas of the upper urinary tract. This proliferative lesion of the urothelium can precede, accompany or follow superficial or invasive urothelial tumours. Rigorous treatment and surveillance are therefore justified to detect an associated tumour. The controversy concerning the benign nature of inverted papilloma has still not been clarified.


[Health economics: should it concern the health sector?].
This paper addresses the most important features of health economics, especially its scope and applications within the sphere of health. Health economics is a field of study which allows countries to gear their health policies toward making more rational use of their resources and expanding and improving their health care services. Such policies should, on the other hand, aim to generate strategies for adequately managing human, technical, economic, and financial resources so as to reap the finest health benefits possible. A knowledge of how economics can be applied to health will enable health professionals to introduce an economics culture into their daily work. In other words, it will allow them to keep in mind the scope of their various working tools--health planning, national health accounts and accounting, and economic assessment methods, which include health and pharmaceutical technology assessment and pharmacoeconomics--as well as the place that economics has in health research. Hopefully, a knowledge of these aspects of economic analysis will provide decision-makers with one more tool they can apply in selecting more efficient options and attaining the highest health benefits at the lowest possible cost for the national health system.


[Design of distal radius volar locking plates : Anatomical, surgical and biomechanical aspects].
The operative treatment of unstable distal radius fractures primarily aims for the anatomical reduction of the joint while addressing accompanying injuries. Anatomical reduction, stable fixation and early functional movement of the joint are the three cornerstones of modern treatment concepts of distal radius fractures. Distal radius volar locking plates play a major role in the treatment and rehabilitation of the most commonly occurring fracture in humans. This article outlines the different principles in the current design of available distal radius volar locking plates. The biomechanical aspects, anatomical findings and clinical evaluation that have influenced current design features and trends in new developments of the latest plates are emphasized. This is an ongoing process that is supported through the investigation and feedback of clinical science.


Allogeneic stem cell transplantation for Fanconi Anaemia. Severe Aplastic Anaemia Working Party of the EBMT and EUFAR. European Group for Blood and Marrow Transplantation.
Fanconi anaemia is a hereditary disorder characterised by chromosomal breaks increased by cross-linking agents. Bone marrow transplantation is the treatment of choice when a HLA identical sibling donor has been identified. The use of low-dose cyclophosphamide with thoraco-abdominal irradiation for the conditioning regimen of FA patients has lead to a dramatic improvement of survival, with a long-term survival of 75% at our institution. However, if most patients are completely cured of their haematological disease, there is concern about an increased frequency of secondary tumours, mostly head and neck squamous cell carcinomas of poor prognosis. Results of BMT using alternative donors (HLA mismatched related and unrelated donors) have also improved during the last decade. A better selection of the donor via high-resolution techniques for class-II HLA matching, and more recently the use of T cell depleted grafts are probably the main explanations. Despite a short follow-up and the small number of patients analysed, transplants using HLA matched family cord blood give some promising results. On the other hand, first results with unrelated cord blood remind that this approach is clearly an experimental one that has to be evaluated through international registries and prospective studies. New approaches including autologous stem cell transplantations and gene therapy are currently explored.


Thermal method for continuous measurement of cerebral perfusion.
A new thermal system using constant heating power for continuous measurement of cerebral perfusion is presented. It is designed and implemented for optimal perfusion sensitivity and dynamic response based on heat-transfer analysis of perfused brain tissue with thermistors on the cortical surface. Two matched thermistors are used, one to serve as a perfusion sensor and the other to compensate for the base-line temperature changes. To improve the signal-to-noise ratio of the measurement system, lock-in amplifiers are used to minimise long-term drift and low-frequency noise. Errors in the measurement caused by electrical and thermal fluctuations are tested and analysed. In vitro tests show that the measurement accuracy of temperature change is better than 10(-3) degrees C, and the temperature resolution is even greater. In vivo evaluation confirms that the system is responsive to cerebral perfusion changes associated with sudden changes in mean arterial blood pressure caused by bolus injection of norepinephrine, blood withdrawal and blood infusion. The dynamic response of the system is sufficient to detect the autoregulatory perfusion changes in response to arterial blood pressure alteration and the oscillations of cerebral blood flow.


[Crow-Fukase syndrome associated with Castleman disease showing hypertrophic cranial pachymeningitis and bilateral internal carotid artery occlusion].
A 51-year-old woman was diagnosed as Crow-Fukase syndrome on July 1997, presenting with lymph node swelling, polyneuropathy, hepatomegaly, hypothyroidism, renal dysfunction, edema and skin change. Lymph node swelling and polyneuropathy improved in some degree after chemotherapy. She was admitted to our hospital on march 6, 1998 because of consciousness disturbance, right hemiparesis and non-fluent aphasia after fever and hypotension. The next day of admission, consciousness disturbance, right hemiparesis and non-fluent aphasia disappeared. MR images of the brain revealed low intensity on a T1-weighted image and high intensity on a T2-weighted image in the left parietal lobe. Furthermore, MR images also revealed diffuse hypertrophic dura matter with enhancement by Gd-DTPA, which made the diagnosis of chronic cranial pachymeningitis. The cerebral angiographies showed bilateral internal carotid artery occlusion. The cerebrospinal fluid showed normal cell count, total protein level of 82 mg/dl, and IgG level of 18 mg/dl. Since there has been very few case reports describing intimate relationship between Crow-Fukase syndrome and pachymeningitis, and between carotid occlusion and pachymeningitis, we speculated that the pachymeningitis might be associated with Crow-Fukase syndrome. Furthermore, pachymeningitis might be a cause of her bilateral carotid occlusion. The number of cases of Crow-Fukase syndrome associated with cerebrovascular disease was very rare. This is the first case which had bilateral internal carotid artery occlusion probably caused by chronic cranial pachymeningitis. Therefore, it is necessary to pay attention to cerebrovascular disease when the patient of Crow-Fukase syndrome is associated with pachymeningitis.


HlyA hemolysin of Vibrio cholerae O1 biotype E1 Tor. Identification of the hemolytic complex and evidence for the formation of anion-selective ion-permeable channels.
Hemolysin (HlyA) was concentrated from supernatants of different Vibrio cholerae O1 biotype E1 Tor strains by ammonium sulfate precipitation. The concentration of the toxin in the supernatants and in the precipitates was quantified using its hemolytic activity. The toxin formed a high molecular-mass band (about 220 kDa) on SDS/PAGE while the toxin monomer had a molecular mass of 60 kDa when it was heated. The addition of the E1 Tor hemolysin oligomers, but not that of the monomers, to the aqueous phase bathing lipid bilayer membranes resulted in the formation of ion-permeable channels, which had long lifetimes at small voltages. The hemolysin channel had a single-channel conductance of 350 pS in 1 M KCl. These results defined hemolysin (HlyA) from V. cholerae as a channel-forming component with properties similar to other cytolytic toxins. The long lifetime of the channel suggested that the channel-forming oligomer did not show a rapid association/dissociation reaction. At voltages larger than 50 mV, the hemolysin channel was voltage dependent in an asymmetric fashion dependent on the side of its addition. The single-channel conductance of the hemolysin (HlyA) from V. cholerae O1 biotype E1 Tor channel was a linear function of the bulk aqueous conductance, which suggested that the toxin forms aqueous channels with an estimated minimum diameter of about 0.7 nm. The hemolysin channel of V. cholerae was found to be moderately anion-selective. The pore-forming properties of hemolysin (HlyA) from V. cholerae O1 biotype E1 Tor were compared with those of aerolysin of Aeromonas sobria and alpha-toxin from Staphylococcus aureus. All these cytolytic toxins must probably oligomerize for activity in biological and artificial membranes and form anion-selective channels.


Mobitz type I heart block after pokeweed ingestion.
The leaves of Phytolacca americana (pokeweed), when ingested, typically produce a self-limited but severe gastroenteritis characterized by intense vomiting and frothy diarrhea. Although cardiac effects have been reported in previous cases of pokeweed ingestion, no cardiac toxin has ever been identified in pokeweed. We report the case of a family who ingested raw and/or cooked pokeweed leaves, and 1 member who developed a Mobitz type I heart block associated with vomiting which resolved after i.v. promethazine. This suggests the possibility that some cardiac effects of pokeweed are secondary to the increased vagal tone seen with severe gastrointestinal colic.


Postnatal maturation of rat intestinal membrane: lipid composition and fluidity.
1. We studied the lipid composition and the fluidity of small intestine brush border membrane (BBM) of rats of different age: 'very young' (5-7 weeks old), 'young' (9 weeks old), 'adult' (30 weeks old) and 'old' (85 weeks old). 2. Fluorescence anisotropy, as assessed by 1,6-diphenyl-1,3,5-hexatriene probe (DPH), was increased from very young to adult rats. 3. In agreement with these results the lipid composition in adult animals showed a lower lipid/protein ratio (derived mainly from a lower content of total polar lipids) and an increase of cholesterol esters and sphingomyelin (SM) saturation index. 4. A marked decrease of the order parameter was observed in the 'old' group, accompanied by a decreased cholesterol/phospholipid ratio. 5. The percentage distribution of membrane phospholipids significantly changed during development, but the modifications were not correlated with the anisotropy of DPH.


Hepatitis B core-related antigen levels are associated with response to entecavir and peginterferon add-on therapy in hepatitis B e antigen-positive chronic hepatitis B patients.
Hepatitis B core-related antigen (HBcrAg), a new serum marker, may be useful in monitoring chronic hepatitis B infection. HBcrAg was measured in 175 hepatitis B e antigen-positive patients treated with entecavir (ETV) with or without peginterferon (PEG-IFN) add-on therapy. Decline in HBcrAg was stronger in patients with vs. without combined response (ETV: -3.22 vs. -1.71 log U/mL, p <0.001; PEG-IFN add-on: -3.16 vs. -1.83 IU/mL, p <0.001) and in patients with vs. without hepatitis B surface antigen (HBsAg) response (ETV: -2.60 vs. -1.74 log U/mL, p <0.001; PEG-IFN add-on: -2.38 vs. -2.15 log U/mL, p = 0.31). HBcrAg was associated with combined response (adjusted odds ratio 0.3, 95% confidence interval 0.2-0.5, p <0.001), but was not superior to quantitative HBsAg (qHBsAg).


Acinetobacter calcoaceticus-baumannii complex bacteremia: analysis of 82 cases.
Eighty-two cases of Acinetobacter calcoaceticus-baumannii complex bacteremia were identified during a 33-month period, from November 1993 to July 1996, at the Veterans General Hospital, Taipei. All cases were due to hospital-acquired infections, with 28 cases of polymicrobial bacteremia. Most patients had severe debilitating conditions: 26 had malignancies, 40 required stay in Intensive Care Unit and 17 had undergone major operations. The main predisposing factors included central venous catheterization, endotracheal intubation or tracheostomy, prior antibiotic therapy and prolonged hospitalization. Amikacin, tobramycin, and ceftazidime were the most effective agents in vitro against A. calcoaceticus-baumannii complex. 32 patients (39 %) died during hospitalization, 19 of the cases (23 %) directly attributed to septicemia. Factors that adversely influenced mortality included polymicrobial bacteremia, inappropriate antimicrobial therapy and prior antibiotic treatment. Of particular interest is the fact that none of the patients who did not receive appropriate antimicrobial therapy survived. Early diagnosis and appropriate antibiotic therapy are critical for improving the prognosis of A. calcoaceticus-baumannii complex bacteremia.


Orthopaedic trauma care: a two-tier system?
Orthopaedic trauma requiring surgical admission presents to our hospitals right throughout the week. However, the level of service provided to trauma patients appears to fluctuate with more surgery facilities available during weekday "office-hours" with reduced facilities at the weekend. The National Confidential Enquiry into Peri-operative Deaths (NCEPOD) in 1999 laid down guidelines for orthopaedic trauma surgery in elderly patients clearly stating that no elderly patient requiring an urgent operation should have to wait for more than 24 h once fit for surgery. We see no reason to exclude the younger population from such a directive and have hence applied the same standard of "surgery within 24 h of admission" as our index of appropriate practice. Audit of our consultant delivered performance confirmed that while an average 88% of "weekday service" patients admitted Sunday through Thursday achieved this standard, only an average of 64% of weekend service patients admitted on Friday or Saturday achieved the same standard. The purpose of this report is to increase awareness of what we believe to be a widespread dilemma. The day of the week should not dictate the treatment of the patient.


Low-Temperature Oxidation-Free Selective Laser Sintering of Cu Nanoparticle Paste on a Polymer Substrate for the Flexible Touch Panel Applications.
Copper nanomaterials suffer from severe oxidation problem despite the huge cost effectiveness. The effect of two different processes for conventional tube furnace heating and selective laser sintering on copper nanoparticle paste is compared in the aspects of chemical, electrical and surface morphology. The thermal behavior of the copper thin films by furnace and laser is compared by SEM, XRD, FT-IR, and XPS analysis. The selective laser sintering process ensures low annealing temperature, fast processing speed with remarkable oxidation suppression even in air environment while conventional tube furnace heating experiences moderate oxidation even in Ar environment. Moreover, the laser-sintered copper nanoparticle thin film shows good electrical property and reduced oxidation than conventional thermal heating process. Consequently, the proposed selective laser sintering process can be compatible with plastic substrate for copper based flexible electronics applications.


Lacosamide monotherapy for partial onset seizures.
To evaluate the 1-year efficacy and safety of oral lacosamide as conversion monotherapy in adult patients with partial onset seizures with or without generalization. We prospectively followed-up consecutive patients converted to lacosamide monotherapy after 1-year seizure freedom on lacosamide add-on therapy and withdrawal of the concurrent antiepileptic drug (AED). Seizure occurrence, treatment compliance and drug toxicity were assessed every 3 months up to 1 year. The study outcomes were the retention rate of lacosamide as single AED and the seizure freedom under lacosamide monotherapy at 1 year from withdrawal of background AED. The safety variable was the prevalence of lacosamide related adverse events (AEs). Among the 58 included patients, at 1 year from withdrawal of background medication, 37 (63.8%) retained lacosamide as single AED and 32 (55.2%) were free from seizure occurrence under lacosamide monotherapy throughout the entire follow-up. The history of less than three lifetime AEDs turned out to be significant predictor of seizure freedom (adjusted OR = 6.38, 95% CI 1.85-21.98, p = 0.003). Twelve (20.8%) subjects reported mild to moderate AEs, with the commonest being drowsiness, dizziness, and headache. Conversion to lacosamide monotherapy could be effective and well tolerated in selected adults patients with partial onset seizures who had achieved seizure freedom during lacosamide add-on therapy.


Glyceraldehyde-3-phosphate dehydrogenase mRNA. Activity and amount in dystrophic hamster muscle.
The activity and amount of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in muscle of young dystrophic hamsters was reduced to approximately half the level found in control animals. No changes in brain or liver enzyme activity were found. Several other glycolytic enzyme activities and creatine kinase activity in muscle were unchanged, except for modest decreases in aldolase and pyruvate kinase. To assess the synthesis of glyceraldehyde-3-phosphate dehydrogenase, the poly(A)+ RNA was isolated from muscle polysomes of dystrophic and control animals and its activity was assessed in an mRNA-dependent translation system. The translatability of the mRNA for GAPDH found in the dystrophic muscle preparations also was half of that found in the control muscle preparations. Decreases were also found in the translatability of mRNA for tropomyosin.


Mechanisms for Kir channel inhibition by quinacrine: acute pore block of Kir2.x channels and interference in PIP2 interaction with Kir2.x and Kir6.2 channels.
Cardiac inward rectifier potassium currents determine the resting membrane potential and contribute repolarization capacity during phase 3 repolarization. Quinacrine is a cationic amphiphilic drug. In this work, the effects of quinacrine were studied on cardiac Kir channels expressed in HEK 293 cells and on the inward rectifier potassium currents, I(K1) and I(KATP), in cardiac myocytes. We found that quinacrine differentially inhibited Kir channels, Kir6.2 ∼ Kir2.3 > Kir2.1. In addition, we found in cardiac myocytes that quinacrine inhibited I(KATP) > I(K1). We presented evidence that quinacrine displays a double action towards strong inward rectifier Kir2.x channels, i.e., direct pore block and interference in phosphatidylinositol 4,5-bisphosphate, PIP(2)-Kir channel interaction. Pore block is evident in Kir2.1 and 2.3 channels as rapid block; channel block involves residues E224 and E299 facing the cytoplasmic pore of Kir2.1. The interference of the drug with the interaction of Kir2.x and Kir6.2/SUR2A channels and PIP(2) is suggested from four sources of evidence: (1) Slow onset of current block when quinacrine is applied from either the inside or the outside of the channel. (2) Mutation of Kir2.3(I213L) and mutation of Kir6.2(C166S) increase their affinity for PIP(2) and lowers its sensitivity for quinacrine. (3) Mutations of Kir2.1(L222I and K182Q) which decreased its affinity for PIP(2) increased its sensitivity for quinacrine. (4) Co-application of quinacrine with PIP(2) lowers quinacrine-mediated current inhibition. In conclusion, our data demonstrate how an old drug provides insight into a dual a blocking mechanism of Kir carried inward rectifier channels.


Head and neck lymphedema management: Evaluation of a therapy program.
The purpose of this pilot study was to examine a therapeutic intervention for head and neck lymphedema. The 22-week intervention involved therapist-led care and participant self-management. Effectiveness was evaluated using a previously described lymphedema assessment tool, the Assessment of Lymphedema of the Head and Neck (ALOHA) to detect change over the course of the 22 weeks of treatment, and before and after a single treatment session. A prospective observational pilot study was conducted with a cohort of 10 participants assessed. Measurements of size (tape measurements) and water content (tissue dielectric constant [TDC]) were used, per the ALOHA protocol. Participants received 13 lymphedema therapy treatments at reducing frequencies over 22 weeks and daily self-management. There was an overall significant reduction in lower neck circumference (F [2.15,19.35] = 7.11; P = .004), upper neck circumference (F [5,45] = 7.27; P < .001) and TDC (F (5,45) = 8.92; P < .001) over time. There were no significant differences over the course of treatment for mean ear-to-ear measurements or before and after a single session of treatment. This pilot study found a reduction in head and neck lymphedema over the 22-week lymphedema treatment course. This intervention may be successful in reducing head and neck lymphedema; however, further studies are needed to investigate these findings in a larger sample with the use of a control group to negate improvements from healing over time.


Present Role of Mammography/Digital Mammography in Breast Cancer Management.
Mammography is the primary breast imaging modality. Its major role is in breast cancer screening. Other indications of mammography are referred to as "diagnostic mammography," and these include evaluation of patients with symptomatic breast diseases, search of the occult primary cancer, preoperative evaluation, and follow-up of patients with breast cancer. Other imaging modalities compliment, but do not generally replace, mammography. Digital mammography has further improved the technical capabilities of mammography.


[Cow's milk protein allergy].
Cow's milk protein allergy affects gastrointestinal morphology and function. The clinical symptoms depend on the localisation in the gut and on the type of the allergic reaction. In some patients cutaneous, respiratory and even urinary symptoms may also be present. As the condition is almost limited to the first year of life, transitory elimination of cow's milk from the diet is the treatment of choice. A number of patients with intractable diarrhoea suffers from undiagnosed cow's milk protein allergy.


Monitoring Shigella flexneri vacuolar escape by flow cytometry.
Invasive bacterial pathogens such as Shigella flexneri force their uptake into non-phagocytic host cells. Upon internalization, they rupture the endocytic vacuole and escape into the host cell cytoplasm. Recent studies applying fluorescence resonance energy transfer (FRET) based methods to track host-pathogen interactions have provided insights into the process of bacterial infection at the single cell level. We have previously reported that the vacuolar escape of invasive bacteria into the host cellular cytosol can be tracked by fluorescence microscopy using a FRET CCF4/β-Lactamase reporter assay. Here, we show that our vacuolar rupture assay can also be analyzed by flow cytometry constituting an important alternative to data acquisition by microscopy. Whereas analysis of our assay by fluorescence microscopy offers precise spatiotemporal resolution, flow cytometry analysis represents a high-throughput method that allows efficient and fast quantification of a large number of events and can further improve future research on vacuolar escape.


Stable, intense picosecond pulse generation using intracavity GaAs.
A reliable pulsed Nd:YAG laser system has been designed to produce 150-mJ pulses of 260-ps duration with a shot-to-shot standard deviation of ±5.8% and a full-angle divergence of 0.6 mrad. The stability is due to a new cavity-dumped oscillator design that uses an intracavity GaAs plate to stabilize the pulse development and inhibit large fluctuations. The resulting output pulse from the cavity-dumped oscillator is 103 µJ, with an average shot-to-shot standard deviation of ±1.1% and a full-angle divergence of 2 mrad.


Applying metabolomic analyses to the practice of embryology: physiology, development and assisted reproductive technology.
The advent of metabolomics technology and its application to small samples has allowed us to non-invasively monitor the metabolic activity of embryos in a complex culture environment. The aim of this study was to apply metabolomics technology to the analysis of individual embryos from several species during in vitro development to gain an insight into the metabolomics pathways used by embryos and their relationship with embryo quality. Alanine is produced by both in vivo- and in vitro-derived human, murine, bovine and porcine embryos. Glutamine is also produced by the embryos of these four species, but only those produced in vitro. Across species, blastocysts significantly consumed amino acids from the culture medium, whereas glucose was not significantly taken up. There are significant differences in the metabolic profile of in vivo- compared with in vitro-produced embryos at the blastocyst stage. For example, in vitro-produced murine embryos consume arginine, asparagine, glutamate and proline, whereas in vivo-produced embryos do not. Human embryos produce more alanine, glutamate and glutamine, and consume less pyruvate, at the blastocyst compared with cleavage stages. Glucose was consumed by human blastocysts, but not at a high enough level to reach significance. Consumption of tyrosine by cleavage stage human embryos is indicative of blastocyst development, although tyrosine consumption is not predictive of blastocyst quality. Similarly, although in vivo-produced murine blastocysts consumed less aspartate, lactate, taurine and tyrosine than those produced in vitro, consumption of these four amino acids by in vitro-derived embryos with high octamer-binding transcription factor 4 (Oct4) expression, indicative of high quality, did not differ from those with low Oct4 expression. Further application of metabolomic technologies to studies of the consumption and/or production of metabolites from individual embryos in a complete culture medium could transform our understanding of embryo physiology and improve our ability to produce developmentally competent embryos in vitro.


Qualitative analysis of the deposit of collagen in bladder suture of rats treated with tacrolimus combined with mycophenolate-mofetil.
To evaluate the synthesis of type I (mature) and type III (immature) collagen in bladder suture of rats treated with a combination of tacrolimus and mycophenolate mofetil for 15 days. Thirty rats were divided into 3 groups: the sham, control and experimental groups. All the animals underwent laparotomy, cystotomy and bladder suture in two planes with surgical PDS 5-0 thread. The sham group did not receive treatment. The control group received saline solution, and the experimental group received 0.1mg/kg/day of tacrolimus with 20mg/kg/day of mycophenolate mofetil, for 15 days. From then on, the tacrolimus was dosed. The surgical specimens of the bladder suture area were processed so that the total type I and type III collagen could be measured by the picrosirius red technique. There was a predominance of type I collagen production in the sham and control groups compared to the experimental group, in which type III collagen was predominant. The production of total collagen did not change. The association of tacrolimus and mycophenolate mofetil in animals qualitatively changes the production of collagen after 15 days with a predominance of type III collagen.


Non-optical screening platforms: the next wave in label-free screening?
The use of optical biosensors for compound screening was first demonstrated in the mid-1990s, but there has been limited uptake in the market owing to issues of limited throughput and a lack of applications for key receptor classes. Recently, several start-up and established tools companies have exploited non-optical detection modalities that seek to address the shortcomings of more established optical approaches. Platforms based on acoustic resonance, electrical impedance, microcantilevers, nanowires and differential calorimetry are beginning to appear with commercially available products targeted at post-high-throughput screening hit confirmation and mode-of-action studies. This article highlights key advances in commercial label-free analysis platforms, which complement more traditional optical system and which also allow novel assay formats for the analysis of previously intractable targets.


[Endospore formation by Streptomyces avermitilis in submerged culture].
The capability of streptomycetes to form endospores during their life cycle was studied in submerged cultures of Streptomyces avermitilis. Submerged S. avermitilis spores were most intensely formed (1) during the culture development cycles on synthetic medium CP1 with glucose under phosphate limitation, and (2) in autolysing cell suspensions of high density obtained by tenfold concentration of a stationary-phase culture grown in a synthetic medium resuspended in phosphate buffer (pH 7.2) with 0.2% CaCl2. Endospores of S. avermitilis formed in submerged cultures shared the major characteristics of specialized microbial resting forms: heat resistance, resistance to lysozyme, ability to pertain to the main species-defining features, and ultrastructural organization characteristic of endospores. They can be considered a resting form of streptomycetes alternative to the spores formed exogenously on aerial mycelium in a surface culture.


Fatty acid composition of serum, adipose tissue, and liver in rats fed diets containing corn oil or cottonseed oil.
There have been few studies on the effect of cottonseed oil (CSO), one of the most commonly used vegetable oils in the United States, on indices of lipid status either in the rat or in any other species. Previous studies with rats have focused on the effect of CSO, versus that of other vegetable oils, on the concentration of cholesterol in serum and liver or on a limited number of fatty acids in serum, adipose tissue, or liver. The present study was designed to compare more fully than in previous studies the effect of CSO to that of corn oil (CO) on the fatty acid composition of serum, adipose tissue, and liver. Two groups of male rats (n = 10 each) were used, each fed one of two purified diets containing 100 g/kg of either CO or CSO. Between-group differences included higher values (mol% total fatty acids) in adipose tissue for palmitic acid and stearic acid, but lower values in serum, adipose tissue, and liver for cis-vaccenic acid and oleic acid (C18:1n-9) for the CSO group than the CO. In addition, values in serum, adipose tissue, and liver for alpha-linolenic acid (C18:3n-3) and its metabolite, docosahexaenoic acid (C22:6n-3), were lower for the CSO group than the CO. There was no effect of diet on serum or tissue levels of n-6 fatty acids (linoleic acid, dihomo-y-linolenic acid, and arachidonic acid). Differences in fatty acid compositions noted, e.g., those for C18:ln-9 and C22:6n-3, mainly reflected compositional differences between the two oils. Thus, use of CSO rather than CO affected serum and tissue concentrations of saturated, monounsaturated, and n-3 polyunsaturated fatty acids (PUFAs) but had no effect on n-6 PUFA status.


Specific up-regulation of DNA polymerase by human papillomavirus 16.
To analyze how the infection of human papillomavirus 16 (HPV16) affects expression of DNA polymerase beta (DNA polB) with the aim of probing the mechanism of over-expression of DNA polB in human cancers. Four fragments of human DNA polB promoter were amplified and constructed into luciferase reporter vector pGL-Basic, generating pGL-BP, pGL-BMH, pGL-BMS, and pGL-BAT constructs respectively, and co-transfected with HPV16 or HPV6 into Hep2 cells. Luciferase activity was assayed 48 hours after transfection. Semi-quantitative RT-PCR was used to measure mRNA expression of endogenous DNA polB. Immunohistochemistry and in situ hybridization were used to analyze DNA polB expression and HPV16 or HPV6 infection in 38 cases of cervical lesions respectively. With co-transfection of HPV16 and DNA polB promoter-driving reporters into Hep2 cells, pGL-BP reporter in full-length DNA polB promoter presented markedly elevated luciferase activities (P < 0.05). However, the other three mutant reporters: pGL-BMH, pGL-BMS, and pGL-BAT, generated no reporting activities in the presence of HPV16 (P > 0.05). On the contrary, all of polB promoter reporters were little stimulated in co-transfection of HPV6 (P > 0.05). The transfection of HPV16 could enhance the endogenous polB mRNA expression compared with that of HPV6 (3.42 vs. 0.80, P < 0.05). The DNA polB expression was found in 8 of 10 HPV16-positive cervical intraepithelial neoplasia grade III (CIN III) cases, while was only found in 3 of 11 HPV6-positive condyloma accuminatum cases, but was negative in all chronic cervicitis cases. The correlation of DNA polB expression with HPV16 infection in cervical lesions was significant (P < 0.05). HPV16 is able to specifically stimulate the expression of DNA polB in human epithelial cells through interaction with the core upstream regulatory sequences of DNA polB promoter. Over-expression of DNA polB might be an explanation for the molecular mechanism underlying HPV-related human cancers.


[Hereditary forms of colorectal adenomatous polyposis].
Hereditary colorectal adenomatous polyposis syndromes are a predisposition to colorectal carcinoma development. The familial adenomatous polyposis is the most common analyzed syndrome that results from germline mutations in the APC gene. In addition to, the autosomal recessive form of polyposis has been recently reported. This disease is caused by germ-line mutations in the base excision repair MYH gene. The goal of this study is the identification of genetic causes of the colorectal polyposis, the determination of the frequency and type of the APC and MYH germ-line mutations in the set of families with colorectal polyposis in Czech population. The set of 103 probands with FAP was screened for germ-line APC mutations using the Protein Truncation Test and Denaturing Gradient Gel Electrophoresis. The MYH mutational screening was performed on 60 unrelated patients without detected APC mutations using the Denaturing High Performance Liquid Chromatography. Automated sequencing was carried out to identify found mutations. Totally, the 51 germ-line APC mutations (69,9%) are reported in the set of 72 probands including 31 novel mutations unique for Czech population. Molecular genetic analysis of the MYH gene revealed 15 DNA variations (25 %) including two patients identified as p.Y 165C/p.G382D compound heterozygotes (3,3%) and 13 polymorphisms or intronic changes (21,7%). The novel variants were detected in the 5 patients. Present study reflects the extremely heterogenous spectrum of the APC mutations in Czech population and confirms the previously reported data. However, the changes found in the MYH gene still need more extensive studies. Our results are important for genetic counselling and further clinical management among at-risk family members. It also enables distinction among different types of the colorectal polyposis.


[Systemic side effects of external corticoids: results of dermatologic studies in children with neurodermititis].
Of 111 children with atopic dermatitis who were admitted as in-patients in a German aero-biological childrens clinic, 83 patients (74.8%) had been prescribed a prolonged corticosteroid therapy from their family doctor. For 60 children (54.1%) the corticosteroid application was topital, partly with occlusive dressing or with fluorinated preparations. The remaining 23 children (20.7%) were treated topically and systemically with corticosteroids. Of the 60 children whose atopic dermatitis was treated exclusively with topical corticosteroids, 59 patients (98.3%) showed undesirable side effects. Moreover, irreversible body changes developed through the transcutaneous absorptive processes, which were more evident and appeared more often in girls than in boys.


The emerging role of SHANK genes in neuropsychiatric disorders.
The genetic heterogeneity of neuropsychiatric disorders is high, but some pathways emerged, notably synaptic functioning. A large number of mutations have been described in genes such as neuroligins, neurexins, and SHANK that play a role in the formation and the maintenance of synapses. This review focuses on the disorders associated with mutations in SHANK3 and the other members of its family, SHANK1 and SHANK2. SHANKs are scaffolding proteins of the postsynaptic density of glutamatergic synapses. SHANK3 has been described in the Phelan-McDermid syndrome (PMS), but also in autism spectrum disorders (ASD) and schizophrenia associated to moderate to severe intellectual disability (ID) and poor language. The evolution of patients with PMS includes symptoms of bipolar disorder and regression. SHANK2 has been identified in patients with ASD with mild to severe ID. SHANK1 has been associated with high-functioning autism in male patients, while carrier females only display anxiety and shyness. Finally, based on neuropathological findings in animal models and patients, a possible role of SHANK in Alzheimer's disease is discussed. Altogether, this review describes the clinical trajectories associated with different mutations of the SHANK genes and provides information to further investigate the role of the SHANK genes in neuropsychiatric disorders.


Counter current line deflection immunoelectrophoresis. A technique for the quantification of antibodies.
Combining the electrophoretic principles of counter current immunoelectrophoresis and deflection of line precipitates in line immunoelectrophoresis provides a new technique for quantitative determination of antibodies against specific antigens (CCLD electrophoresis), even if no purified antigen or monospecific antibodies are available for construction of the detection system. We have used the method for quantitative determination of antibodies against the flagellum of Treponema phagedenis biotype Reiter and compared the diagnostic potential of this method in the diagnosis of syphilis with an ELISA method for the quantification of IgG antibodies against the flagellum. The CCLD electrophoresis could be optimized to a diagnostic performance very similar to that achieved using the ELISA method.


Substituent Dependent Optical Properties of p-phenyl Substituted ethenyl-E-thiophenes.
Various electron donor and acceptor substituted (NO2, CN, Cl, H, OCH3, NH2) p-phenyl ethenyl-E- thiophenes (1-6) were synthesized and substituent dependent optical properties (dipole moment, transition dipole moment, oscillator strength, optical band gap, hyperpolarizability) were studied using Solvatochromism and Density functional theory. It is shown that thiophene acts as a weak electron donor in presence of an electron withdrawing p-phenyl substituent (NO2, CN, Cl), whereas thiophene acts as a weak electron acceptor in presence of an electron donating p-phenyl substituent (OCH3, NH2). In comparison to ethenyl thiophene 4, the HOMO-LUMO energy band gap is decreased upon increasing the electron donating or electron withdrawing capacity of p-phenyl substituent. From the excited state dipole moment calculation, it is shown that the excited state is highly dipolar for nitro and amino compounds 1 and 6, whereas compounds 2-5 show a non-polar excited state. As compared to the ethenyl thiophene 4, the first hyperpolarizability (β) increases upon substitution either with a strong electron withdrawing or strong electron donating p-phenyl substituent. A large β value is found for p-nitro phenyl ethenyl-E-thiophene and p-amino phenyl ethenyl-E- thiophene. Overall, these studies provide useful information in understanding the optical properties of phenyl and heterocyclic based ethenyl systems.


The Longitudinal Interval Follow-up Evaluation. A comprehensive method for assessing outcome in prospective longitudinal studies.
The Longitudinal Interval Follow-up Evaluation (LIFE) is an integrated system for assessing the longitudinal course of psychiatric disorders. It consists of a semistructured interview, an Instruction booklet, a coding sheet, and a set of training materials. An interviewer uses the LIFE to collect detailed psychosocial, psychopathologic, and treatment information for a six-month follow-up interval. The weekly psychopathology measures ("psychiatric status ratings") are ordinal symptom-based scales with categories defined to match the levels of symptoms used in the Research Diagnostic Criteria. The ratings provide a separate, concurrent record of the course of each disorder initially diagnosed in patients or developing during the follow-up. Any DSM-III or Research Diagnostic Criteria disorder can be rated with the LIFE, and any length or number of follow-up intervals can be accommodated. The psychosocial and treatment information is recorded so that these data can be linked temporally to the psychiatric status ratings.


[Affected Factors of Separating Erythrocytes by Discontinuous of Density Gradient Centrifugation].
To study the affected factors of the separation of red blood cells by discontinuous density gradient method, and to establish the optimal operating conditions of percoll for the separation of erythrocytes. The osmolality of different ratio of Percoll and 1.5 mol/L NaCl were 9:1, 10:1, 11:1, 12:1, 13:1 ratio, was detected respectively. The isotonic osmolality ratio was selected as optimum stock isotonic Percoll solution(SIP). Evaluation criteria: All the different density cells were separated sufficiently by centrifugation, there is little red cell retention and entrainment phenomena. The separation efficacy of different red cell suspension concentration, different centrifugal force, different centrifugal speed, different temperature and different time were compared. The separation efficacy of red cell suspension concentration which not exceed 50% was better than above 50%, efficacy of 4 000×g was superior to 3 000×g and 2 000×g, 2-8 centrifugal acceleration and deceleration centrifugal could obtain the satisfactory efficacy, the 4 °C layered effect was more stable than room temperature, the separation effect of 20 min was better than 15 min and 10 min. Percoll and 1.5 mol/L NaCl 12:1 to 11:1 can be used as stock isotonic percoll solution, preferentially selected 4 °C as centrifugal conditions. When conditions of separation was satisfied, the centrifugal acceleration and deceleration is 2-8, the optimal centrifugal effect can be obtained with the decrease of erythrocyte suspension concentration, with centrifugation time prolonging and centrifugal force increasing.


Human surrogate models of histaminergic and non-histaminergic itch.
Within the last decade understanding of the mechanistic basis of itch has improved significantly, resulting in the development of several human surrogate models of itch and related dysesthetic states. Well-characterized somatosensory models are useful in basic studies in healthy volunteers, in clinical studies for diagnostic and segmentation purposes, and in pharmacological studies to evaluate the antipruritic efficacy of existing and novel compounds. This review outlines recently introduced histamine-independent human models of itch, their mechanisms, their ability to induce clinically relevant phenomena, such as alloknesis, and the results obtained through their use. The article also introduces recent advances in the understanding of itch and provides an overview of the methods to assess experimentally-induced itch and associated manifestations. Major improvements are warranted in the treatment of chronic pruritus, and reliable human surrogate models are a valuable tool in achieving them, both for basic researchers and for clinicians.


Board's eye view - Guiding principles.
I heard a daft comment on morning-time television the other day. Apparently, the reason school nurses do not put plasters on children's bleeding wounds is because wounds heal better when they have dried. The statement made me so cross I almost sent a text message of complaint to the broadcaster.


High permissivity of the fish cell line SSN-1 for piscine nodaviruses.
Seventeen isolates of piscine nodavirus from larvae or juveniles of 13 marine fish species affected with viral nervous necrosis (VNN) were examined for their infectivity to a fish cell line SSN-1. Based on cytopathic effects (CPE) and virus antigen detection by fluorescent antibody technique (FAT) after incubation at 25 degrees C, the infectivity of these virus isolates was divided into 4 groups. Group 1, including 9 virus isolates from 4 species of grouper, 2 species of sea bass, barramundi, rock porgy, and Japanese flounder showed CPE characterized by rounded, granular cells with heavy cytoplasmic vacuoles within 3 d post-incubation (p.i.), and the monolayer partially or completely disintegrated over 3 to 6 d p.i. Scattered FAT-positive cells appeared at 3 h p.i. and spread through the cell sheet with an increasing fluorescence signal over 24 h p.i. Group 2, consisting of 3 virus isolates from striped jack, induced CPE with thin or rounded, granular, refractile cells without conspicuous vacuole formation, and extensive FAT-positive reaction was observed in a time course similar to that of Group 1. Cells inoculated with Group 3 (1 isolate from tiger puffer) developed no distinct CPE but viral infection was evidenced by localized FAT-positive cells. There were no FAT-positive cells in Group 4, which included 4 isolates from Japanese flounder, Pacific cod and Atlantic halibut. However, when incubation was performed at 20 degrees C, the SSN-1 cells inoculated with the Group 3 isolate showed CPE similar to that of Group 1 and extensive FAT-positive reaction. Evidence of virus proliferation at 20 degrees C was also obtained in Group 4 isolates. The virus titers in the infected fish varied from 10(11) to 10(16) tissue culture infectious dose (TCID50) g(-1) of fish. There is a good correlation between these infectivities to the SSN-1 cells and the coat protein gene genotypes of the isolates. The present results indicate that SSN-1 cells are useful for propagating and differentiating genotypic variants of piscine nodavirus.


Student perceptions of osteopathic manipulative treatment after completing a manipulative medicine rotation.
Recent studies have demonstrated a decline in the use of osteopathic manipulative treatment (OMT) by osteopathic physicians, reflecting a trend that may begin in medical school. The authors used a questionnaire to examine the teaching and use of OMT in five rotations and the perceptions of 86 graduating osteopathic medical students of their experiences following their core manipulative medicine rotation. Most students indicated that they applied osteopathic principles sometimes (39.5%) or often (29.1%) during rotations. Forty-three percent of students rated their ability to apply osteopathic principles as average. The number of students who indicated that they rarely used OMT during their rotations was 31 (36.0%) for internal medicine, 21 (24.4%) for surgery, 23 (26.7%) for pediatrics, and 24 (27.9%) for obstetrics/gynecology. When asked why OMT was not used during a rotation, 47.2% of respondents cited time constraints, and 21.7% stated that their attending physicians discouraged the use of OMT. These results demonstrate a distinction between students' perceived level of osteopathic principles and skills and their application during clinical rotations.


Wheeler and Whitehead: Process Biology and Process Philosophy in the Early Twentieth Century.
The case of William Morton Wheeler and Alfred North Whitehead represents a striking example of how biologists and philosophers engaged in a common enterprise in the early twentieth century. Both challenge the notion that the living world is composed of distinct organisms. Based on his studies of the behavior of social insects, Wheeler developed a concept of superorganisms that paved the way for a theory of emergent evolution. This paper argues that Whitehead, whose relation to academic biology has been largely ignored, drew on Wheeler's findings and integrated them into a universal philosophical cosmology.


Secreted enzymes of Aeromonas.
A hallmark characteristic of species of Aeromonas is their ability to secrete a wide variety of enzymes associated with pathogenicity and environmental adaptability. Among the most intensively studied are beta-lactamases, lipases, hemolytic enterotoxins, proteases, chitinases, nucleases and amylases. Multiple copies of genes encoding each type of enzyme provide additional biological diversity. Except for the chitinases, these multiple copies show little evolutionary relatedness at the DNA level and only limited similarity at the protein level. Indeed a number of the genes, such as nuclease H of A. hydrophila, have no similarity to known prokaryotic or eukaryotic sequences. The challenge is to determine how these genes evolved, where they originated and why Aeromonas possesses them in such abundance and variety.


Ultrastructural postmortem changes in chicken kidneys at 27 C.
Ultrastructural postmortem changes of glomeruli, proximal tubules (PT), distal tubules (DT), and cortical collecting ducts (CD) were studied in adult chickens held at 27 C for 0, 1, 5, 10, 30, 60, or 360 minutes. There were marked ultrastructural alterations in tissues considered normal by light microscopy. The earliest changes, evident in immediately fixed samples, consisted of apical cell swelling of PT and DT epithelium and formation of smaller projections from glomerular capillary endothelium. Swelling increased with time, and gaps in cell membranes of tubular cytoplasmic blebs were evident by 10 minutes postmortem. The urinary space also contained cellular debris in immediately fixed samples and was filled by swollen epithelial cells by 10 minutes postmortem. Organelle changes were pronounced by 5 minutes in PT epithelium and by 10 minutes in DT epithelium. These initial changes consisted of dilation of mitochondrial and endoplasmic reticulum. Flocculent mitochondrial matrix densities were present by 5 minutes in PT epithelium and enlarged and appeared in other tubular segments as the time postmortem increased. Nuclear chromatin margination occurred first in 10-minute samples of PT and DT, and it was present by 30 minutes in glomerular epithelial cells. Chromatin clumping was present in PT and DT epithelium at 30 minutes and was followed by pyknosis and karyorrhexis at 360 minutes. These latter nuclear changes were not found in glomerular cells during the study. Nuclei of CD showed little change during the periods studied. The glomerular basement membrane appeared as a single hyaline membrane by 30 minutes postmortem, but this membrane and podocyte foot processes showed little further change by 360 minutes postmortem.


Interference of IP-10 expression inhibits vascular smooth muscle cell proliferation and intimal hyperplasia in carotid artery: a new insight in the prevention of restenosis.
After vascular angioplasty, vascular smooth muscle cell (VSMC) proliferation causes atherosclerosis and intimal hyperplasia leading to restenosis. Interferon-γ-inducible protein (IP)-10 plays a role in atherogenesis, but the mechanism remains unclear. We evaluated the role of IP-10 in intimal hyperplasia and restenosis. IP-10 expression was determined in arterial specimens from 20 arteriosclerotic obliteration patients and 6 healthy individuals. VSMCs were stimulated in vitro with IFN-γ and transfected with IP-10 siRNA. Silencing was verified with RT-PCR/Western blot; cell proliferation rate was detected by methyl-thiazol-tetrazolium. The carotid artery model of atherosclerosis injury was established with IP-10 siRNA. IP-10 expression was detected at 1 and 4 weeks using RT-PCR and immunohistochemistry. Artery morphology was assessed with hematoxylin-and-eosin staining, and intimal hyperplasia was evaluated by electron microscopy. IP-10 was overexpressed in arteriosclerotic obliteration group compared with control group (P < 0.05). IP-10 expression in transfected group was significantly lower than in untransfected group. The intima-to-media ratio of transfected group at 4 weeks was lower than that of untransfected group (P < 0.01). The transfected group exhibited more regular intimal structure and less hyperplasia under electron microscopy. We, therefore, concluded that IP-10 played an important role in intimal hyperplasia as siRNA-mediated IP-10 silencing inhibited aberrant VSMCs hyperplasia and reduced restenosis.


Serum soluble transferrin receptor in hypochromic microcytic anaemia.
The objective of this study was to assess the clinical significance of soluble transferrin receptor (sTfR) in hypochromic microcytic anaemia. Serum sTfR was determined on 91 blood samples with haemoglobin less than 110 g/L and MCV less than 76 fl or MCH less than 27.0 pg. The samples were classified as iron deficiency anaemia (IDA), anaemia of chronic disease (ACD) and thalassaemia. ACD was further divided into groups 1 and 2, based on the serum ferritin level. The sTfR level in the control was 1.53+/-0.8 mg/L. In IDA, the sTfR level was 5.53+/-8.76 mg/L and this was significantly higher compared with the control (p-value is less than 0.0001). sTfR levels in ACD (3.32+/-3.94 mg/L) and the thalassaemia group(1.64+/-1.02 mg/L) were not significantly different from that of the control (p-value is more than 0.05). In ACD, the sTfR level in group 1 was significantly higher when compared with the control (p-value is less than 0.001) and group 2 (p-value is less than 0.01). A significantly higher sTfR/ferritin ratio was observed in IDA (2,368.98+/-7,236.4 microg/microg) compared to the control (37.6+/-43.7 microg/microg) (p-value is less than 0.001). No significant difference was noted between ACD, thalassaemia and control (p-value is greater than 0.05). sTfR/ferritin ratio was significantly higher in group 1 of ACD when compared with that of control and group 2 (p-value is less than 0.001). Serum sTfR and sTfR/ferritin ratio are useful parameters in hypochromic microcytic anaemia to diagnose iron deficiency particularly when associated with chronic inflammation. sTfR can be done selectively in ACD patients when ferritin levels are more than 60 microg/L and there is a diagnostic dilemma. If sTfR level is raised, a trial of iron therapy is suggested for these patients.


The military gynecologist in low-intensity conflict environment.
U.S. military medical personnel have been assigned to Honduras since August 1983. While reserve components have 2-week assignments and Air Force medics receive 3-month assignments, U.S. Army medical personnel are assigned for 6-month tours. Duties include rendering medical care and air evacuation capability to all U.S. Forces in the Central American area of operations, conducting medical civic action programs termed MEDRETES, and assisting the host government in developing and conducting a health care system for indigent rural Hondurans. Gynecological care to women soldiers assigned to this low-intensity conflict environment and gynecological surgery capability for indigent Hondurans has been one important aspect of the military medical presence.


The application of quality improvement tools to increase the number of day-only admissions: a rural hospital's experience.
Hospital discharge data were analysed and showed Tamworth Base Hospital as having fewer day-only admissions compared to similar hospitals. A team of ten staff involved in all aspects of day-only patient transactions was selected to analyse work processes associated with the day-only patient system with the aim of increasing the number of day-only admissions. From the application of quality improvement tools, the team was able to make recommendations to increase the work flow and predict resources necessary to increase day-surgery throughput. A revised system allowing for an increase in the number of admissions has been introduced; this has seen a significant increase in the number of people admitted as day-only patients. A telephone survey following discharge of day-surgery patients indicates that standards of care have been maintained and improved.


Coarse-grained protein model with residue orientation energies derived from atomic force fields.
Coarse-grained models for protein simulations can potentially access longer time scales in larger protein systems than atomic-level models. Here, a coarse-grained residue pair potential, with distance and orientation dependency, is derived from equilibrium ensembles of residue pairs generated by molecular dynamics (MD). In particular, the Boltzmann inversion method is used to determine the energies. The residue pair potential is combined with local dihedral angle potentials for the backbone and side chains and used in the folding simulations of six small proteins, (28-67 residues) containing a variety of secondary structures. For the proteins tested, folding simulations by Monte Carlo methods generate structures similar to the native ones. However, these native-like structures were among the lowest in energy for alpha helical proteins but not for proteins containing extended beta structures. It is also found that a careful balance between local and nonlocal interactions is essential. Possibilities for improving coarse-grained models derived from atomic force fields are discussed.


An antifungal naphthoquinone, xanthones and secoiridoids from Swertia calycina.
A chemical and biological screening of 25 species of the Gentianaceae family has been undertaken. Both methanolic and dichloromethane extracts of Swertia calycina exhibited a strong antifungal activity against Cladosporium cucumerinum and Candida albicans. The compound responsible for this activity has been isolated and identified as 2-methoxy-1,4-naphthoquinone. It is the first naphthoquinone to be described in Gentianaceae species. LC-UV and LC-TSP-MS analysis of the crude extracts of Swertia calycina also allowed on-line identification of six known xanthones and secoiridoids.


May zoledronic acid have negative effects on cognition and muscle performance?
We aimed to investigate the effects of zoledronic acid treatment on daily living activities, cognitive functions, depression, muscle strength, and performance. The study was conducted retrospectively. Bone mineral densitometry (BMD) values, Katz activities of daily living (ADL), Lawton-Brody instrumental activities of daily living (IADL), mini mental state examination (MMSE), geriatric depression scale (GDS), mini nutritional assessment (MNA), grip strength, and gait speed scores before and 6 months after zoledronic acid administration were compared. A total of 115 patients were included in the study. There was a significant increase in lumbar total (p < .001), femoral neck (p = .002), and femur total (p = .001) BMD values after zoledronic acid treatment. Significant decrease was found in MMSE (p = .016) and gait speed scores (p = .008) after zoledronic acid treatment, but no significant difference was found in terms of Katz ADL, Lawton-Brody IADL, MNA, GDS, and grip strength (p > .05). Our study indicated that zoledronic acid did not affect daily living activities, depression, and muscle strength. Although we have concluded that cognitive and muscle performance may be adversely affected by zoledronic acid treatment.


Space-Confined Polymerization: Controlled Fabrication of Nitrogen-Doped Polymer and Carbon Microspheres with Refined Hierarchical Architectures.
The construction of refined architectures plays a crucial role in performance improvement and application expansion of advanced materials. The synthesis of carbon microspheres with a refined hierarchical structure is still a problem in synthetic methodology, because it is difficult to achieve the necessary delicate control of the interior structure and outer shell across the microscale to nanoscale. Nitrogen-doped multichamber carbon (MCC) microspheres with a refined hierarchical structure are realized here via a surfactant-directed space-confined polymerization strategy. The MCC precursor is not the traditional phenolic resol but a new kind of 2,6-diaminopyridine-based multichamber polymer (MCP) with a high nitrogen content up to 20 wt%. The morphology and sizes of MCP microspheres can be easily controlled by a dual-surfactant system. The as-synthesized MCC with a highly microporous shell, a multichamber inner core, and beneficial N-doping can serve as a promising supercapacitor material.


Effect of sample numbers on the kinetic data analysis of MR contrast agents.
The Monte Carlo simulation method is used to determine the uncertainties of the estimated are and mean transit time (MTT) of a regional cerebral concentration-time curve of a bolus injected paramagnetic contrast agent. Many factors contribute to these uncertainties. This study concentrates on the error propagation from the sample noise at different sample time points along the concentration-time curve. The uncertainties of area and MTT were determined as a function of the sample noise and the number of sample time points. For example, the percentage standard deviations (uncertainties) of both area and MTT are less than 12% with five samples whose signal-to-noise ratios are greater than 50.


Validation of the 16-Gene Recurrence Score in Patients with Locoregional, High-Risk Renal Cell Carcinoma from a Phase III Trial of Adjuvant Sunitinib.
Purpose: Adjuvant sunitinib prolonged disease-free survival (DFS; HR, 0.76) in patients with locoregional high-risk renal cell carcinoma (RCC) in the S-TRAC trial (ClinicalTrials.gov NCT00375674). The 16-gene Recurrence Score (RS) assay was previously developed and validated to estimate risk for disease recurrence in patients with RCC after nephrectomy. This analysis further validated the prognostic value of RS assay in patients from S-TRAC and explored the association of RS results with prediction of sunitinib benefit.Patients and Methods: The analysis was prospectively designed with prespecified genes, algorithm, endpoints, and analytical methods. Primary RCC was available from 212 patients with informed consent; primary analysis focused on patients with T3 RCC. Gene expression was quantitated by RT-PCR. Time to recurrence (TTR), DFS, and renal cancer-specific survival (RCSS) were analyzed using Cox proportional hazards regression.Results: Baseline characteristics were similar between patients with and those without RS results, and between the sunitinib and placebo arms among patients with RS results. RS results predicted TTR, DFS, and RCSS in both arms, with the strongest results observed in the placebo arm. When high versus low RS groups were compared, HR for recurrence was 9.18 [95% confidence interval (CI), 2.15-39.24; P < 0.001) in the placebo arm; interaction of RS results with treatment was not significant.Conclusions: The strong prognostic performance of the 16-gene RS assay was confirmed in S-TRAC, and the RS assay is now supported by level IB evidence. RS results may help identify patients at high risk for recurrence who may derive higher absolute benefit from adjuvant therapy. Clin Cancer Res; 24(18); 4407-15. ©2018 AACR.


Misuse of stimulant medication among college students: a comprehensive review and meta-analysis.
The misuse of stimulant medication among college students is a prevalent and growing problem. The purpose of this review and meta-analysis is to summarize the current research on rates and demographic and psychosocial correlates of stimulant medication misuse among college students, to provide methodological guidance and other ideas for future research, and to provide some preliminary suggestions for preventing and reducing misuse on college campuses. Random-effects meta-analysis found that the rate of stimulant medication misuse among college students was estimated at 17 % (95 % CI [0.13, 0.23], p < .001) and identified several psychological variables that differentiated misusers and nonusers, including symptoms of attention-deficit/hyperactivity disorder, problems associated with alcohol use, and marijuana use. A qualitative review of the literature also revealed that Greek organization membership, academic performance, and other substance use were associated with misuse. Students are misusing primarily for academic reasons, and the most common source for obtaining stimulant medication is peers with prescriptions. Interpretation of findings is complicated by the lack of a standard misuse definition as well as validated tools for measuring stimulant misuse. The relation between stimulant medication misuse and extra curricular participation, academic outcomes, depression, and eating disorders requires further investigation, as do the reasons why students divert or misuse and whether policies on college campuses contribute to the high rates of misuse among students. Future research should also work to develop and implement effective prevention strategies for reducing the diversion and misuse of stimulant medication on college campuses.


[Hippocrates's evaluation of the madness of Democritus].
Hippocrates is a historical as well as a legendary character. In legends he is presented as a personification of the highest intellectual and moral virtues as they were understood by the ancient greeks and romans. One of these legends deals with a fictitious meeting between him and Democritus, the philosopher. We can follow the story in a collection of pseudepigraphic letters from the end of the first christian century. On the behalf of the senate of the city of Abdera Hippocrates takes upon himself the commission to cure Democritus - the eminent son of the city - who is supposed to be mad because he is laughing at everything. However, Hippocrates soon realizes that his commission transcends his medical profession. Democritus is not mad but a very wise man whose yearn for solitude and serenity constrains him to discriminate himself from ordinary people by an odd behaviour. He is a melancholic. In these letters we follow a discussion about moral and medical aspects of madness and the relationship between the noble outsider and his ordinary fellowmen. It is a discussion that points forward at recurrent themes in the history of ideas. Hippocrates stands out as a philantropist and Democritus as a misanthropist.


[Focusing monodrama. Individual psychodrama treatment: the treatment concept, treatment method of the therapist using psychodrama and focusing technics--a case report].
Moreno's psychodrama is useful in group as well as in individual therapy. Individual psychodrama is known as monodrama. The following paper presents the idea of "focusing monodrama". The concepts of treatment procedure, course of therapeutic session, and the therapist's mode of working will be specifically examined. A one case study exemplifies the treatment procedure in focusing monodrama.


Engineered human Tmpk fused with truncated cell-surface markers: versatile cell-fate control safety cassettes.
Cell-fate control gene therapy (CFCGT)-based strategies can augment existing gene therapy and cell transplantation approaches by providing a safety element in the event of deleterious outcomes. Previously, we described a novel enzyme/prodrug combination for CFCGT. Here, we present results employing novel lentiviral constructs harboring sequences for truncated surface molecules (CD19 or low-affinity nerve growth factor receptor) directly fused to that CFCGT cDNA (TmpkF105Y). This confers an enforced one-to-one correlation between cell marking and eradication functions. In-vitro analysis demonstrated the full functionality of the fusion product. Next, low-dose 3'-azido-3'-deoxythymidine (AZT) administration to non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice injected with transduced clonal K562 cells suppressed tumor growth; furthermore, one integrated vector on average was sufficient to mediate cytotoxicity. Further, in a murine xenogeneic leukemia-lymphoma model we also demonstrated in-vivo control over transduced Raji cells. Finally, in a proof-of-principle study to examine the utility of this cassette in combination with a therapeutic cDNA, we integrated this novel CFCGT fusion construct into a lentivector designed for treatment of Fabry disease. Transduction with this vector restored enzyme activity in Fabry cells and retained AZT sensitivity. In addition, human Fabry patient CD34(+) cells showed high transduction efficiencies and retained normal colony-generating capacity when compared with the non-transduced controls. These collective results demonstrated that this novel and broadly applicable fusion system may enhance general safety in gene- and cell-based therapies.


Quantification and cellular localization of ovine placental lactogen messenger ribonucleic acid expression during mid- and late gestation.
Ovine placental lactogen (oPL) is structurally similar to PRL, is a product of the chorionic epithelium, and has been implicated in playing a supportive role in fetal growth. This study examined the concentration and cellular location of oPL mRNA at five stages of pregnancy (days 60, 90, 105, 120, and 135) in 21 cross-bred ewes, and results were compared to maternal and fetal serum oPL concentrations, cotyledonary DNA and actin mRNA concentrations, and total fetal weight. The concentration of oPL mRNA in fetal cotyledonary tissue increased (P < or = 0.05) from day 60 (15.4 pg/micrograms total cellular RNA) to day 120 (73.7 pg/micrograms total cellular RNA) of gestation and then plateaued, whereas no significant changes occurred in the concentration of actin mRNA over the gestational ages examined. The concentration of DNA in cotyledonary tissue (micrograms per mg wet tissue) increased (P < or = 0.05) from days 60 through 120 and remained constant through day 135, such that when oPL mRNA was expressed on a picogram per microgram DNA basis, no stage of gestation effect (P > or = 0.10) was observed. The maternal serum oPL concentration increased (P < or = 0.05) from day 60 (7.1 ng/ml) to day 105 (417.7 ng/ml), followed by a large but nonsignificant (P > or = 0.10) increase in maternal serum oPL occurring on day 135 (902.0 ng/ml). Fetal serum oPL concentrations increased (P < or = 0.05) from day 60 (11.0 ng/ml) to day 90 (29.0 ng/ml) and then remained relatively constant. Maternal serum oPL (r = 0.68; P < or = 0.01) and cotyledonary oPL mRNA levels (r = 0.61; P < or = 0.05) were correlated with total fetal weight when adjusted for fetal number and gestational age, and together accounted for 80.6% (r2 value) of the variation found in total fetal weight. The correlation between fetal serum oPL concentrations and total fetal weight was nonsignificant (P < or = 0.10). Examination of placentome cross-sections by immunocytochemistry and in situ hybridization at the five gestational ages indicated that the chorionic binucleate cell was the sole source of oPL. These data provide evidence that, like maternal serum concentrations of oPL, oPL mRNA expression by chorionic binucleate cells increases until late gestation, whereas fetal serum concentrations of oPL plateau during midgestation.(ABSTRACT TRUNCATED AT 400 WORDS)


Interleukin-6 up-regulates the expression of interleukin-15 is associated with MAPKs and PI3-K signaling pathways in the human keratinocyte cell line, HaCaT.
Interleukin (IL)-15 is an important inflammatory cytokine and plays a key role in autoimmune disease. At present, IL-15 gene expression and regulation related to many innate immunity trigger signals have been clarified in some specific cell types, but the relationship of IL-6 and IL-15 in the human keratinocyte cell line (HaCaT) is unknown. In this study, we investigated the effect of IL-6 on the expression of IL-15 and selected signaling pathways in HaCaT cells. Results demonstrated that IL-6 up-regulated the expression of IL-15 both at the mRNA and protein levels. Meanwhile, IL-6 was able to activate MAPKs-ERK1/2 and PI3K-AKT signaling pathways. Furthermore, the high expression of IL-15 induced by IL-6 was down-regulated while MAPKs-ERK1/2 and PI3K-AKT signaling pathways were, respectively, blocked by PD98059 and LY294002. These findings indicate that the expression of IL-15 up-regulated by IL-6 is associated with MAPKs-ERK1/2 and PI3K-AKT signaling pathways in HaCaT cells.


[Description of patients diagnosed with "alcohol dependence syndrome" and "alcohol abuse" according to the C.I.E.-9a criteria of the W.H.O. at a psychiatric hospitalization service].
The authors make an introduction where they describe several problems found in the alcoholic diseases researches. The most important of them are related to the "alcoholic case" concept. After this the authors report several epidemiological facts about the alcoholic diseases and disorders in Castilla-León and Madrid. Then a retrospective analysis of inpatients psychiatric records from the Psychiatry Department of Valladolid's University Hospital between 1980 to 1984 (N = 1.259) is made. 252 (20.01%) of them suffered an alcoholic disorders, following the I.C.D.-9th criteria psychoses alcoholic were 102 (8.10%) and they were excluded. However "Alcohol dependence" with 128 cases (10.17%) and "Alcohol Abuse Syndromes" with 22 cases (1.75%) were studied deeply, being 150 cases--120 males (80%) and 30 females (20%)--evaluated. Every diagnostic group was analysed in a distinct way through 41 epidemiological, clinical, diagnostic and medical care variables. These two groups and their differences and similarities between them are reported. The main profile is a male patient, 43 years old, married, urban life, blue collar worker, unskilled work, primary studies, bad economic level, marriage and working maladapted, clinical admitted through the Emergency Service, with an alcoholism 24 years old and intake of 234 grs. alcohol/day, with former treatment of minor tranquilizers (B.D.Z.) and at last is readmitted by alcoholic disease of disorder or a different psychiatric disease or disorders (I.C.D.-9.a or D.S.M.-III) in a 37.5% and 16.41% respectively.


Retrospective analysis of microorganisms isolated from cystic fibrosis patients in Southern Italy, 2002-2010.
This study aim was to determine the prevalence of microorganisms in the respiratory tract of patients with cystic fibrosis (CF) admitted to the CF Reference Centre in Southern Italy between 2002-2010. Microbiology assessment of samples (sputum and tracheal aspirates) collected from patients with pulmonary exacerbation admitted to hospital was carried out. All patients were registered in a database and clinical and microbiological data were retrospectively analysed. Overall, 188 patients were included and a total of 1217 samples were analysed. The most common microorganisms were Staphylococcus aureus (78.7% of the patients) and Pseudomonas aeruginosa (58%), followed by Candida albicans (19.1%), Haemophilus influenzae (13.3%) and Aspergillus fumigatus (9.6%). Compared to similar studies performed in other European countries, our microbiological data, especially the low occurrence of filamentous fungi, suggest a specific local epidemiology, probably related to some uncommon CFTR mutations, which are specific to Southern Italy.


Demonstration of antigenic heterogeneity of Neisseria gonorrhoeae pili antigens using human sera in the test system.
Antigenic heterogeneity of gonococcal pili using human sera in the test system was demonstrated by means of indirect haemagglutination (IHA) inhibition studies. The reactivities of sera from 104 females with current gonococcal infection were compared by the IHA test using different pili preparations as antigens. Sera and gonococcal strains were collected from 365 patients. Pili were purified from strains infecting patients who met the following three requirements: 1) no history of previous gonococcal infection(s), 2) an infection that had lasted a week or more, 3) a seronegative result when tested by the IHA test using pili from Buchanan's strain 2686 as antigen. Each patient's serum was tested against pili produced by the gonococcal strain infecting this particular patient. A seropositive reaction obtained by using pili antigen from the strain infecting the seronegative (strain 2686) patient indicates an antigenically heterogeneous pili preparation. Two such strains, antigenically different from one another and from strain 2686, were found among 22 pili preparations investigated.