Clinical relevance of associated aneurysms with arteriovenous malformations of the posterior fossa.
The aim of this study was to determine the frequency of aneurysms associated with arteriovenous malformations (AVMs) of the posterior fossa and their relation to hemorrhagic presentation in comparison to supratentorial AVMs. We performed a retrospective analysis of 474 patients with intracranial AVMs treated in our center from 1990 to 2010. Patients were analyzed for AVM size, drainage type and their clinical course with focus on vessel anomalies including AVM-associated aneurysms. Seventeen (30%) of 57 posterior fossa AVMs versus 46 (11%) of 417 supratentorial AVMs were associated with aneurysms. In six (10.5%) versus seven (1.7%) patients, respectively, flow-associated aneurysms were the source of hemorrhage. Infratentorial location of an AVM was a significant risk factor for the incidence (p < 0.001) and rupture (p < 0.001) of AVM-associated aneurysms. Feeding artery aneurysms in particular represented a risk factor for hemorrhage in the overall group of AVM patients, independently of the location (p < 0.001). The majority of patients with a posterior fossa AVM were treated by combined embolization and surgical removal within one procedure (n = 33, 58%). Feeding artery aneurysms were excluded by endovascular coiling or surgical clipping whenever feasible. Overall treatment-associated permanent morbidity in the subgroup of posterior fossa AVMs was 11% (n = 6) and mortality 4% (n = 2). Posterior fossa AVMs display a significantly higher frequency of associated aneurysms of the adjacent vessels that are correlated to the high bleeding rate compared to AVMs of the supratentorial compartment. We therefore recommend aggressive AVM treatment including the exclusion of associated aneurysms as a minimal therapeutic goal whenever possible.


Surgical and functional outcomes of sigmoid vaginoplasty among patients with variants of disorders of sex development.
To assess the use of sigmoid colon in vaginal reconstruction of some patients with disorders of sex development. The study included 31 patients with disorders of sex development of various causes. All were reared as females. Female gender was decided for all cases after complete medical assessment. All patients underwent sigmoid vaginoplasty. Assessment of surgical and functional outcomes was carried out in a follow-up period of up to 6 years. The preoperative diagnoses included mullerian aplasia (16 cases), androgen insensitivity syndrome (12 cases) and previous failed vaginoplasty (3 cases). Associated surgical procedures were gonadectomy in 5 cases and gonadectomy combined with clitoroplasty and vulvoplasty in 7 cases. No intra-operative or early postoperative complications occurred. A cosmetic neovagina with adequate size was achieved in all cases. Long term follow-up showed introital stenosis in 4 cases (12.9 %). Two of them responded to vaginal dilatation. The third one needed y-v plasty while the fourth one presented by acute abdomen secondary to ruptured vagina and was submitted to urgent laparotomy. Mucosal prolapse occurred in 1 case (3.2 %). Reoperation rate was 9.6 %. Sexual satisfaction was achieved among 9 sexually active cases. The subjective satisfaction score of the surgical outcome was 8.03. For patients with disorders of sex development of various etiologies, sigmoid vaginoplasty is the preferred technique for vaginal replacement. It is a safe technique that provides the patient with a cosmetic neovagina of adequate caliber and a satisfactory functional outcome.


[Medicamentous inhibitory therapy of precancerous lesions of the esophagus--3 and 5 year inhibitory effect of antitumor B, retinamide and riboflavin].
9633 subjects aged 40 to 65 were sampled from high-risk areas for esophageal cancer and examined by esophageal exfoliative cytology. 2531 and 3393 cases of markedly and mildly dysplastic patients were screened out respectively. Those with marked dysplasia were randomized into 3 groups and given respective medications: antitumor B (Chinese herbs), 4-ethoxycarbophenylretinamide (retinamide) and placebo. The subjects with mild dysplasia were randomly divided into 2 groups for riboflavin and placebo treatment. After medication for 3 or 5 years the subjects were reexamined by esophageal exfoliative cytology. The incidence of esophageal cancer in the antitumor B group after taking medication for 3 and 5 years was reduced by 52.2% and 47.3% respectively as compared with the placebo group. These differences were statistically significant (chi 2 = 8.9115, P less than 0.01; chi 2 = 10.9573, P less than 0.01). The incidence of esophageal cancer in the retinamide group after 3 years of medication was reduced by 37.3% as compared with control, while the incidence among patients treated for 5 years dropped by 43.2% relative to control. This difference was statistically significant (chi 2 = 9.2836, P less than 0.01). The incidence of esophageal cancer in the riboflavin group was reduced by 22.2% and 34.8% respectively. The above results indicate that secondary prevention of esophageal cancer is possible and that inhibitory therapy of precancerous lesions of the esophagus is effective in preventing esophageal cancer. This method needs further trial and study in high risk areas of esophageal cancer. The reliability of the experimental results is critically discussed.


[Video-controlled, retroperitoneoscopic, lumbar sympathectomy and sympathicotomy].
Six patients with arterial occlusions of the legs underwent VRLS: a small incision is made in the flank, the retroperitoneum pushed aside with a 1.5-1-balloon catheter, the lumbar sympathicus endoscopically severed and/or partially excised with video assistance. The intervention was successful in all cases. Excised parts of the sympathetic trunk were demonstrated histologically in four cases. One case of severe bleeding from a lumbar artery was stopped endoscopically, and one patient underwent additional open resection of 2 cm of the sympathetic trunk. Postoperative intestinal paralysis lasted a mean of two days, and hospitalization five days. VRLS has several disadvantages for the surgeon, while its lesser strain and earlier rehabilitation are a definite advantage for the patient.


Psychological aspects of genetic counseling. VIII. Suffering and countertransference.
Two common forms of countertransferential problems seen in genetic counseling, associative and projective, are described and illustrated. Both forms have the potential of reducing the quality of empathy counselors provide counselees. When counselors experience the same problems as the counselees they counsel it is virtually impossible to avoid facing countertransferential reactions which may not always promote the counselees' interests and/or redound to their benefit. Genetic counselors, like other personal counselors and psychotherapists, have a professional responsibility to be aware of, monitor, contain, and learn from their countertransferential experiences.


Female infanticide and human sex ratio evolution.
The possible effect on the evolution of the human sex ratio of a preference for male children is examined in a genetic model. It is shown that the killing of infant daughters can lead to either a female- or a male-biased sex ratio, the outcome depending on the decision rule used to determine the fate of a child. This reconciles long-standing contradictory results.


Second-Guessing Scientists and Engineers: Post Hoc Criticism and the Reform of Practice in Green Chemistry and Engineering.
The article examines and extends work bringing together engineering ethics and Science and Technology Studies, which had built upon Diane Vaughan's analysis of the Challenger shuttle accident as a test case. Reconsidering the use of her term "normalization of deviance," the article argues for a middle path between moralizing against and excusing away engineering practices contributing to engineering disaster. To explore an illustrative pedagogical case and to suggest avenues for constructive research developing this middle path, it examines the emergence of green chemistry and green engineering. Green chemistry began when Paul Anastas and John Warner developed a set of new rules for chemical synthesis that sought to learn from missed opportunities to avoid environmental damage in the twentieth century, an approach that was soon extended to engineering as well. Examination of tacit assumptions about historical counterfactuals in recent, interdisciplinary discussions of green chemistry illuminate competing views about the field's prospects. An integrated perspective is sought, addressing how both technical practice within chemistry and engineering and the influence of a wider "social movement" can play a role in remedying environmental problems.


Cost-effectiveness of interventions to enhance medication adherence in psychiatric patients: a systematic review.
It was conducted a systematic review of economic evaluations that assessed the cost-effectiveness of interventions to enhance the medication adherence in psychiatric patients. Several bibliographic databases were searched: MEDLINE, MEDLINE in process, PSYCINFO, EMBASE, Cochrane Controlled Trials Register, CINAHL, CRD, EconLit, Science Citation Index and Social Science Citation Index. Full economic evaluations which assessed interventions to enhance the adherence to drug therapy in adult patients with a mental illness were included. Data were extracted and the methodological quality of selected studies was assessed. The information was synthesized through narrative procedures. Four clinical trials and two ongoing studies fulfilled the selection criteria. Two studies did not find significant differences in adherence between the interventions (a compliance-enhancing program, a therapeutic drug monitoring and a pharmacy-based intervention) and the control groups; one study found that a compliance program was more effective than a non-specific counselling intervention over 18 months in psychotic patients; another study found better results in terms of adherence among high-severity depressed patients receiving a stepped collaborative care during 12 months in comparison to a control group; in moderate-severity patients the differences disappeared after the first 6 months. None of the four studies found significant differences in costs between groups because of the low statistical power. In summary, adherence enhancing programs could be cost-effective in psychiatric patients although this statement is based on few studies with limited methodological quality. It is necessary more and better research on the cost-effectiveness of interventions in the field of mental health.


[Enzymatic decomposition of pectin in the presence of amino acids and proteins].
The influence of amino acids--glycine, alpha-alanine, lysine, phenylalanine, serine, methionine, cysteine, tryptophan, aspartic acid and asparagine--and proteins--chick egg albumin and autolysate of Aspergillus niger biomass--on the disintegration of beet pectine by the Asp. niger enzymic preparation was investigated. This process was shown to depend on the chemical composition and concentration of amino acids and the protein nature. All the tested amino acids, except tryptophan, at doses of 0.0001 to 0.5% stimulated the activity of pectolytic enzymes. The effect of proteins was different: Asp. niger autolysate showed no essential influence and egg albumin inhibited pectine disintegration.


Effect of aquatic physical therapy on pain and state of sleep and wakefulness among stable preterm newborns in neonatal intensive care units.
To evaluate the effects of aquatic physical therapy on pain and on the cycle of sleep and wakefulness among stable hospitalized premature infants. This study was characterized as an uncontrolled clinical trial on a time series and included 12 clinically stable newborns of gestational age less than 36 weeks who were hospitalized in a neonatal intensive care unit (NICU). After selection, the newborns were placed in a liquid medium for aquatic physical therapy lasting 10 minutes. Movements to stimulate flexor posture and postural organization were performed. The sleep-wakefulness cycle was assessed using the adapted Brazelton (1973)* scale and pain was assessed by the occurrence of signs of pain according to the Neonatal Facial Coding System (NFCS) scale; and physiological parameters. In relation to states of sleep and wakefulness, before the physical therapy, the newborns' behavior varied from fully awake with vigorous body movements to crying. After the physical therapy, the states of sleep ranged from light sleep with closed eyes to some body movement. These values presented statistically significant differences (p<0.001). The score on the pain assessment scale also decreased from 5.38+/-0.91 to 0.25+/-0.46, with p<0.001 after the intervention. The vital signs remained stable. It is suggested that aquatic physical therapy can be a simple and effective method for reducing pain and improving sleep quality among preterm infants in NICUs. Controlled studies with larger numbers of subjects are needed in order to generalize the results. Article registered of the Clinical Trials under the NCT00785837.


Prevalence of Neisseria Gonorrhoeae among men screened for Chlamydia Trachomatis in four United States cities, 1999-2003.
Neisseria gonorrhoeae infections are the second most commonly reported disease in the United States and cause significant morbidity. We describe the prevalence of gonorrhea in a large sample of men tested for gonorrhea and Chlamydia trachomatis in Baltimore, Denver, San Francisco, and Seattle. Gonorrhea prevalence was measured among 17,712 men tested in a variety of non-sexually transmitted disease (STD) clinic venues using urine-based nucleic acid amplification tests. Among 16,850 asymptomatic men, prevalence ranged from 0% to 1.5% by city (P=0.20): Baltimore 1.3%, Denver 1.5%, San Francisco 1.5%, and Seattle 0%. Among 862 symptomatic men, the gonorrhea prevalence varied from 0.0% to 28.3% by city (P<0.01). The high prevalence of gonorrhea in symptomatic men supports the importance of testing for symptomatic men. The prevalence of gonorrhea among asymptomatic men is low, and routine screening cannot be recommended when screening is performed for chlamydia, unless a substantial local prevalence of gonorrhea can be documented in specific targeted venues or population groups.


Community changes following shrub invasion of grassland.
We studied the development of the shrub Baccharis pilularis ssp consanguinea and its effects on herbs of the annual grassland in Northern California. A series of shands of Baccharis was sampled of ages ranging from 1 yr to>9 yr, representing most of the life cycle of the shrub. In each stand we examined shrub biomass, structure and litterfall. We also determined cover and biomass of all herbaceous species and estimated seed production, seed rain and storage of seed in the soil. Abundances of all herbaceous species declined greatly after Baccharis formed a closed canopy at 2-3 yr, and little seed of herbaceous species was either dispersed into shrub stands or stored in the soil. Exclosures suggested that herbivory by small mammals in the closed shrub stands may be important in reducing the abundance of herbaceous species following shrub invasion of grassland.


Building undergraduate nursing students' knowledge of the research process in nursing.
This article describes a simple approach to teaching a nursing research course to undergraduate nursing students which was received with enthusiasm by both students and faculty. Through short answer progressive assignments throughout the semester involving students in the thinking processes required at each stage of the research process, student knowledge of the research process was progressively developed. Students chose a topic of personal interest for a proposed study and carried this topic through the various phases of the research process. Students positively evaluated this approach frequently commenting that the approach helped them to "think in different ways than before" and strongly recommended continuance of the approach in future course presentations.


Treatment of symptomatic convergence insufficiency with home-based computerized vergence system therapy in children.
To evaluate the efficacy of a home-based computer orthoptic program for symptomatic convergence insufficiency (CI) in children. The medical records of participants aged 5 to <18 years who were diagnosed with symptomatic CI and were treated with the Computerized Vergence System (CVS) program were retrospectively reviewed. All participants were prescribed 9 or 15 minutes of daily convergence exercises with the CVS program, 5 days per week, for the initial 6 weeks. Near point of convergence (NPC) and near convergence amplitude (NCA) were measured at baseline, 6 week, and final examinations. The presence or absence of diplopia and asthenopia with reading were recorded at baseline and final examinations. A total of 186 participants were included. At diagnosis, 72 participants (39%) reported diplopia and 182 (98%), reported asthenopia. At final examination, 172 participants (92%) were asymptomatic. Twelve participants (6%) subsequently received other treatment modalities. Mean NPC at baseline (5.9 cm) improved after 6 weeks of CVS therapy (3.3 cm) and at final examination (2.9 cm; P < 0.0005). Mean NCA at baseline (20.3(Δ)) improved after 6 weeks of CVS therapy (37.0(Δ)) and at the final examination (38.0(Δ); P < 0.0005). In this study, home-based CVS therapy reduced symptoms related to CI and improved the NPC and NCA of most children aged 5 to <18 years with symptomatic CI.


Effect of the quorum-sensing luxS gene on biofilm formation by Enterococcus faecalis.
Enterococcus faecalis is the species of bacterium most frequently isolated from the root canals of teeth that exhibit chronic apical periodontitis refractory to endodontic treatment. In this study, we evaluated the effect of the S-ribosylhomocysteine lyase (luxS) quorum-sensing gene on E. faecalis biofilm formation by constructing a knockout mutant. The biofilms formed by both E. faecalis and its luxS mutant strain were evaluated using the MTT method. Important parameters that influence biofilm formation, including cell-surface hydrophobicity and the nutrient content of the growth medium, were also studied. Biofilm structures were observed using confocal laser scanning microscopy (CLSM), and expression of biofilm-related genes was investigated using RT-PCR. The results showed that the luxS gene can affect biofilm formation, whereas it does not affect the bacterial growth rate. Deletion of the luxS gene also increased cell-surface hydrophobicity. Biofilm formation was accelerated by the addition of increasing concentrations of glucose. The CLSM images revealed that the luxS mutant strain tends to aggregate into distinct clusters and relatively dense structures, whereas the wild-type strain appears confluent and more evenly distributed. All genes examined were up-regulated in the biofilms formed by the luxS mutant strain. The quorum-sensing luxS gene can affect E. faecalis biofilm formation.


Readers' Questions.
TOPICAL ANALGESICS IN LEG ULCER PATIENTS.


Pregnancy loss in the antiphospholipid-antibody syndrome--a possible thrombogenic mechanism.
The mechanisms of vascular thrombosis and pregnancy loss in the antiphospholipid-antibody syndrome are unknown. Levels of annexin V, a phospholipid-binding protein with potent anticoagulant activity, are markedly reduced on placental villi from women with this syndrome. Hypercoagulability in such women may therefore be due to the reduction of surface-bound annexin V by antiphospholipid antibodies. To test this idea, we studied how antiphospholipid antibodies affect levels of annexin V on cultured trophoblasts and human umbilical-vein endothelial cells and how they affect the procoagulant activity of these cells. We isolated IgG fractions from three patients with the antiphospholipid-antibody syndrome and from normal controls. These antibodies were incubated with cultured BeWo cells (a placental-trophoblast cell line), primary cultured trophoblasts, and human umbilical-vein endothelial cells. Annexin V on the cell surfaces was measured by an enzyme-linked immunosorbent assay. The coagulation times of plasma overlaid on the cells were also determined. Trophoblasts and endothelial cells exposed to antiphospholipid-antibody IgG as compared with control IgG had reduced levels of annexin V (trophoblasts, 0.37 +/- 0.02 vs. 0.85 +/- 0.12 ng per well, P=0.02; endothelial cells, 1.6 +/- 0.04 vs. 2.1 +/- 0.05 ng per well, P=0.001). Also, trophoblasts and endothelial cells exposed to antiphospholipid-antibody IgG had faster mean (+/- SE) plasma coagulation times than cells exposed to control IgG (trophoblasts, 8.7 +/- 2.0 vs. 21.3 +/- 2.9 minutes, P=0.02; endothelial cells, 9.8 +/- 0.8 vs. 14.2 +/- 1.2 minutes, P=0.04). Antiphospholipid antibodies reduce the levels of annexin V and accelerate the coagulation of plasma on cultured trophoblasts and endothelial cells. The reduction of annexin V levels on vascular cells may be an important mechanism of thrombosis and pregnancy loss in the antiphospholipid-antibody syndrome.


[Intraoperative and early postoperative complications of puncture-dilation tracheostomy in neurosurgical patients].
The paper provides a detailed analysis of the most common and clinically significant complications due to percutaneous dilation tracheostomy performed in 479 patients with neurosurgical diseases in 2002-2007. It also considers such complication of the technique, which is specific to neurosurgical patients, as elevated intracranial pressure (ICP), and describes a procedure of tracheostomy in patients with elevated ICP. Percutaneous dilation tracheostomy made by a skilled surgeon is shown to be effective and safe.


[The functional condition of patients following a cerebrovascular accident].
Most patients who suffer a cerebrovascular accident (CVA) belong to the adult-elderly age group. Thus, such patients are likely to display a certain degree of functional compromise as a result of the natural process of aging, regardless of whether they have suffered a CVA or not. AIM. To examine the functional condition of patients with CVA, as well as that of healthy individuals who have never suffered a stroke. An analytical cross-sectional study was conducted with a control group made up of 82 individuals -41 of whom were CVA patients (G1) and 41 were individuals with no history of CVA (G2)-. These subjects were paired according to sex, age and educational level. The sample was predominantly made up of females (60%), who were married (70%), with ischaemic CVA (70%), in the right hemisphere of the brain (56%) and with a primary education (36%). The prevalence rate of patients with severe functional impairment was 58.53%, whereas in the control group there were no subjects with this level of deficiency. A relationship was found to exist (p < 0.05) between the functional condition and the variables age and causation of CVA for G1, and also between the functional condition and the variable age for the individuals in G2, which indicates that functional compromise is greater in more elderly subjects. Findings suggest that the functional compromise of patients with CVA is higher than that of individuals with no history of CVA with the same sex, age and educational level. This clearly shows that there is an association between CVA and functional impairment in these patients.


Altered patterns of resting-state functional connectivity between the caudate and other brain regions in medication-naïve children with attention deficit hyperactivity disorder.
Structural and functional alterations occur in the caudate of patients with attention-deficit/hyperactivity disorder (ADHD). Here we aimed to investigate the functional connectivity between the dorsal caudate and other brain regions in ADHD children. Resting-state functional connectivity from 30 ADHD and 33 age- and gender-matched "normal" children were measured by functional Magnetic Resonance Imaging. Positive connectivity with dorsal caudate was observed in the prefrontal areas, cingulate cortex and temporal lobe. Negative functional connectivity was observed in the precuneus, occipital cortices and cerebellum. The connectivity of left dorsal caudate to left inferior frontal gyrus was correlated with severity of ADHD. Connectivity of dorsal caudate with several brain regions was identified in ADHD children.


Statins for Treating Alzheimer's Disease: Truly Ineffective?
Animal studies suggest that statins may have a protective effect on Alzheimer's disease (AD), but some clinical trials have reported negative results. To evaluate the efficacy and safety of statins in the treatment of AD. We searched the PubMed, Cochrane Library, EMBASE, OVID SP, and ProQuest databases to select double-blind, randomized controlled trials pertaining to statins given to patients diagnosed with AD. The meta-analysis used main outcomes such as scores on the Mini Mental State Examination (MMSE) and Alzheimer's Disease Assessment Scale (ADAS-Cog) and secondary outcomes such as scores on the Dependence Scale, Activities of Daily Living Scale (ADCS-ADL), Neuropsychiatric Inventory (NPI), Clinical Global Impression of Change (CGIC), and Alzheimer's Disease Functional Assessment and Change Scale (ADFACS). Safety measures included the standard reporting of any adverse events or laboratory abnormalities. Four studies (1,127 participants) involving patients with a diagnosis of probable or possible AD were included. There were no significant differences between the statins and placebo groups regarding the main outcomes, secondary outcomes, or adverse events. Most of the studies ignored or downplayed risk factors for cerebral vascular disease. Statins are well tolerated without unexpected adverse events. However, more research is needed to determine whether statins are effective for AD with vascular risk factors.


Surface properties of calcium phosphate particles for self setting bone cements.
Calcium phosphate cements (CPC), consist of multicomponent powder mixtures of calcium orthophosphates with grain sizes in the region of 1-20 microm. Due to the small particle sizes surface properties as the zeta potential and adsorption processes play a significant role during manufacturing and application. In the context of this work zeta potentials of different calcium phosphates, like dicalcium phosphate anhydride (DPCA) tetracalcium phosphate (TTCP) and hydroxyapatite were measured in various organic/aqueous media with different pH values. The results show a strong dependency of the zeta potential on the kind of suspension medium used associated with different milling properties. The addition of sodium phosphate leads to a pH value dependent stabilization of the particles in the liquid phase; the zeta potential of the surface increases from about -15 to -18 mV in water and from -35 to -45 mV in 0.05 mol/l sodium phosphate solution. Besides the interaction of particles with various antibiotics was determined on the basis of the zeta potential of the surface. The substances partly cause a tremendous change of the surface load. This is accompanied by a change of the rheological properties of the cement paste, the morphology of the hardened cement matrix and a significant deterioration of the application-relevant properties as setting time or mechanical strength.


Hydrogen bonding between sugar and protein is responsible for inhibition of dehydration-induced protein unfolding.
The nature of the interaction responsible for the inhibition of protein unfolding and subsequent damage by sugars during dehydration is unclear. The relationship between sample moisture content measured by coulometric Karl Fischer titration and the apparent moisture content predicted by the area of the protein side chain carboxylate band at approximately 1580 cm-1 in infrared spectra of dried protein-sugar samples was examined. For samples in which a high level of native protein structure was retained in the dried solid, the apparent moisture content predicted by the carboxylate band area was greater than the actual moisture content, indicating that protection results from direct sugar-protein hydrogen bonding and not entrapment of water at the protein surface. Further, we show that the degree of structural protection conferred by sucrose and trehalose apparent in second derivative, amide I infrared spectra, correlates with the extent of hydrogen bonding between sugar and protein. The failure of dextran to inhibit dehydration-induced lysozyme unfolding is shown to result from the inability of the polymer to hydrogen bond adequately to the protein. Therefore, formation of an amorphous phase alone is not sufficient to maintain protein structure during dehydration. Glucose hydrogen bonds to a high degree with dried lysozyme, but is incapable of inhibiting lyophilization-induced protein unfolding in the absence of an effective cryoprotectant. However, the addition of polyethylene glycol, which is known to protect proteins during freezing, but not drying, to glucose protected lysozyme structure during lyophilization. Together, these results show that hydrogen bonding between carbohydrate and protein is necessary to prevent dehydration-induced protein damage. However, hydrogen bonding alone is not sufficient to protect proteins during lyophilization in the absence of adequate freezing protection.


Distribution of free and sulfate-conjugated catecholamines in human platelets.
Sulfate conjugation represents a major pathway for the inactivation of catecholamines and is presumed to occur at sites, including platelets, where phenylsulfotransferase activity has been identified. This study examines the relative distribution of free and conjugated norepinephrine, epinephrine, and dopamine in the platelets and plasma of resting subjects. As active uptake of free catecholamines into platelets has been reported in vitro; the possibility of a similar mechanism, operating for sulfate conjugates, was also investigated. The presence of sulfated catecholamines in platelets is confirmed. The proportion of total catecholamines existing in the free form was 78 +/- 4% (mean +/- SE) for norepinephrine, 52 +/- 5% for epinephrine, and 50 +/- 10% for dopamine, which was considerably higher than for plasma. Increases in plasma free and conjugated catecholamines achieved by bicycle exercise and oral administration of amines, respectively, were not accompanied by corresponding increases in platelet levels. The origin of platelet conjugates would appear to be the result of intraplatelet sulfation of free amines rather than direct uptake from plasma. Because acute elevation of plasma free and conjugated amines are not reflected in increased platelet levels, it is unlikely that platelet uptake mechanisms play a significantly role in the disposal of circulating catecholamines.


Osteonecrosis of the jaw.
Bisphosphonates are effective therapy for osteoporosis, Paget's disease, and metastatic bone disease. Generally, the side effects of bisphosphonates are minimal. Recently, an uncommon adverse reaction affecting the maxilla or mandible, called osteonecrosis of the jaw, has been reported, especially in those patients receiving high doses of bisphosphonates in the oncology setting. Regarding doses used to treat osteoporosis, clinicians must keep the very small potential absolute risk of jaw osteonecrosis in perspective and consider it in relation to the demonstrated benefit of bisphosphonates. Still, in a very small number of patients taking bisphosphonates, intractable, painful, nonhealing exposed bone may occur following dental extractions or denture irritation.


Long-term potentiation in slices from subcortically deafferented hippocampi.
Long-term potentiation (LTP) can be initiated in CA1, but not in the dentate, region of slices from rats with bilateral subcortical deafferentation at 8 week survival time. At about one week survival time, LTP is present in the CA1 region of all and also in the dentate of about 50% of slices. The results suggest that subcortical cholinergic and catecholaminergic inputs are not required for the LTP in the CA1 area of the rat hippocampus.


Insecticidal activity of five commercial mosquito coils against Anopheles albimanus, Aedes albopictus, and Culex quinquefasciatus.
The insecticidal activity of 4 mosquito coils containing pyrethroid ester insecticides and 1 mosquito coil containing 15% citronella oil was evaluated against adult female Anopheles albimanus, Aedes albopictus, and Culex quinquefasciatus. Two of the pyrethroid mosquito coils containing 0.08% meperfluthrin showed the strongest insecticidal activity, and all 4 pyrethroid coils produced significantly higher mortality than the citronella coil across all 3 test species.


Reproductive medicine meets human embryonic stem cell (hESC) research: the need to adjust the regulatory framework to actual expectations and potential detrimental consequences of hESC research.
Human embryonic stem cell (hESC)-based cell therapy depends on access to surplus embryos from IVF cycles and collaborative interactions between biomedical researchers and reproductive medicine professionals. It is becoming instrumental to achieve an international consensus about the standards that should regulate the manipulation of human embryonic tissue in two distinct settings: reproductive medicine and embryonic stem cell research. Within hESC research, the regulatory framework needs to be adjusted according to the actual expectations and potential detrimental consequences of hESC research.


Biomechanical characteristics of suture techniques in extensor zone IV.
This study was designed to investigate several biomechanical parameters involved in repair of extensor tendons in Zone IV (over the proximal phalanx). Typical suture techniques over the proximal phalanx did not shorten the tendon significantly nor was there significant loss of flexion at the metacarpophalangeal or proximal interphalangeal joints. The Kleinert modification of the Bunnell technique and modified Kessler technique seemed to be the strongest and provided evidence that dynamic or active range of motion, under controlled conditions and in short arcs, might be physiologically tolerated by repaired tendons.


Mammary NOS-type sarcoma with CD10 expression: a rare entity with features of myoepithelial differentiation.
We present an extensive immunohistochemical analysis of 7 mammary sarcomas that did not fit into any specific soft tissue sarcoma category. Histologically, they were composed of spindle cells with highly pleomorphic nuclei and abundant mitoses. Our immunohistochemical antibody panel included pan-cytokeratin (CK), basal cell type CKs (34betaE12, CK5/6, CK14, CK17) and vimentin antibodies, antibodies to established (SMA, CD10, p63, S-100, maspin, calponin, GFAP, SM-myosin), and novel (CD29, 14-3-3sigma) myoepithelial markers, as well as antibodies to CD34, desmin, h-caldesmon, steroid receptors (estrogen, progesterone, androgen), and EGFR (Her-1). Whereas CKs, CD34, desmin, and h-caldesmon were not expressed, all tumors were positive for CD10 and vimentin. CD29 and SMA were observed in 3 cases each (43%), and p63 and calponin in 2 cases each (29%). Other myoepithelial markers and steroid receptors were absent, except androgen receptors, which were expressed in one sarcoma. Five sarcomas showed positivity for EGFR. The distinction of specific, histogenetically defined sarcoma entities (such as leiomyosarcoma, angiosarcoma, liposarcoma) from NOS-type sarcoma with CD10 expression is usually clear-cut because the former exhibit a characteristic histomorphology and immunoprofile. Phyllodes tumors with stromal overgrowth or recurrent phyllodes tumors lacking epithelial structures as well as periductal stromal sarcomas can be ruled out by their frequent expression of CD34 and negativity for myoepithelial markers. The most important differential diagnosis is sarcomatoid metaplastic carcinoma because its treatment includes axillary lymphadenectomy. Since some NOS-type sarcomas with CD10 expression and most metaplastic carcinomas show positivity for CD29, SMA, and p63, differential diagnosis can be extremely difficult and requires extensive immunohistochemical evaluation for CKs and additional myoepithelial markers such as S-100, 14-3-3sigma, and maspin. The immunophenotype of NOS-type sarcomas with CD10 expression suggests that these neoplasms represent a mammary sarcoma variant with myoepithelial features.


[Streptomycin-3"-phosphotransferases from streptomycin-resistant cells of Escherichia coli strains].
Streptomycin-3"-phosphotransferases were isolated and purified from E. coli cells containing plasmids 836, pBS52 or R6K, which determine the microorganisms resistance towards streptomycin and dihydrostreptomycin. Phosphorylation of the 3"-hydroxylic group of dihydrostreptomycin was demonstrated by [13C]-NMR spectrometry. It was shown that streptomycin-3"-phosphotransferase, whose synthesis is determined by plasmid 836 (as well as by plasmid R6K), differs from the analogous enzyme, whose synthesis is operated by plasmid pBS52 in some properties, e. g. dependence of the initial reaction rate on concentrations of antibiotics and ATP, pH-optimum, sensitivity to the buffer ionic strength, stability, etc. Besides, the antiserum against streptomycin-3'-phosphotransferase detected by plasmid pBS52 does not produce cross immunological reactions with the other enzyme.


Anticipating one's troubles: the costs and benefits of negative expectations.
Although negative expectations may have the benefit of softening the blow when a negative event occurs, they also have the cost of making people feel worse while they are waiting for that event to happen. Three studies suggest that the cost of negative expectations is greater than the benefit. In 2 laboratory experiments and a field study, people felt worse when they were expecting a negative than a positive event; but once the event occurred, their prior expectations had no measurable influence on how they felt. These results suggest that anticipating one's troubles may be a poor strategy for maximizing positive affect.


Concomitant use of alcohol and benzodiazepine hypnotics in psychiatric outpatients: a cross-sectional survey.
Concomitant use of benzodiazepines and alcohol appears prevalent in a clinical setting. The objectives of this study were as follows: (1) to investigate the prevalence of concomitant use of benzodiazepine hypnotics and alcohol in psychiatric outpatients, (2) to examine the clinical characteristics and factors associated with the concomitant use, and (3) to investigate the awareness of the psychiatrists-in-charge about the concomitant use. Outpatients with schizophrenia, depression, and insomnia who were receiving benzodiazepine hypnotics were asked to fill in a sleeping diary for seven consecutive days in which use of hypnotics and alcohol was also recorded. Clinical characteristics were assessed, and logistic analysis was performed to examine factors associated with the concomitant use. In addition, psychiatrists-in-charge were asked as to whether they thought their patients were concomitantly using them. The prevalence rate of the concomitant use was 39.8% (37/93). The CAGE score showed significant positive association with the concomitant use (odds ratio = 2.40, 95% confidence interval = 1.39-4.16, P = 0.002). Only in 32.4% of the concomitant users were suspected by their psychiatrists. The results suggest that concomitantly used benzodiazepine hypnotics and alcohol appears prevalent, and has been frequently overlooked by treating psychiatrists. The CAGE questionnaire may be helpful to screen such potentially hazardous users.


Optimization of culture conditions for Gardnerella vaginalis biofilm formation.
Bacterial vaginosis is the leading vaginal disorder in women in reproductive age. Although bacterial vaginosis is related with presence of a biofilm composed predominantly by Gardnerella vaginalis, there has not been a detailed information addressing the environmental conditions that influence the biofilm formation of this bacterial species. Here, we evaluated the influence of some common culture conditions on G. vaginalis biofilm formation, namely inoculum concentration, incubation period, feeding conditions and culture medium composition. Our results showed that culture conditions strongly influenced G. vaginalis biofilm formation and that biofilm formation was enhanced when starting the culture with a higher inoculum, using a fed-batch system and supplementing the growth medium with maltose.


Chronic relapsing thrombotic thrombocytopenic purpura: role of therapy with cyclosporine.
Relapsing thrombotic thrombocytopenic purpura (TTP) is a rare disorder with most individuals experiencing 1 to 5 relapses. We report a patient with 18 episodes of thrombotic thrombocytopenic purpura (TTP), the highest number of relapses thus far described. The last 11 episodes were treated with regimens containing cyclosporine. The patient's medical record was reviewed for pertinent clinical, laboratory, and treatment data. We summarized various parameters for each episode and compared characteristics of relapses treated with vs. without cyclosporine. The initial episode of TTP was unusual in that it failed to respond to plasmapheresis, glucocorticoids, and fresh frozen plasma (FFP). It remitted only following splenectomy. Episodes 2-7 responded to FFP plus prednisone. Episode 8 failed to respond to prednisone plus FFP but remitted promptly with cyclosporine plus prednisone. Subsequently, 2 relapses responded to cyclosporine alone, 2 to cyclosporine plus FFP, 4 to cyclosporine plus prednisone +/- FFP, and 2 to cyclosporine, FFP, prednisone, and plasma exchange. There was no difference in remission duration, or in severity or duration of relapses treated with vs. without cyclosporine. Use of cyclosporine, however, significantly decreased the requirement for prednisone and the length of maintenance therapy; thus it is effective mainly as an adjunctive therapy for TTP.


Assessment of enamel erosion and protective effect of salivary pellicle by surface roughness analysis and scanning electron microscopy.
To assess dental erosion caused by 0.1% and 1.0% citric acid in vitro and to estimate the protective influence of experimentally formed salivary pellicle. Bovine enamel slabs (n = 80) were polished and embedded in epoxy resin. For the formation of pellicle layer 40 specimens were immersed for 24 h in pooled human saliva. Erosion was caused by immersion in citric acid solution for 1, 5, 10 and 30 min. Erosive alterations on the pellicle-covered and non-covered enamel specimens were scored as a change (delta) of surface roughness parameters Ra, Rt and RzDIN using contact profilometer and observed in scanning electron microscope. Profilometric analysis of eroded enamel specimens emphasized the aggressiveness of even low concentrated citric acid with a short period of challenge. The change of roughness parameters after 1-min immersion in 0.1% citric acid were 16.4, 182.6 and 132.2 nm for deltaRa, deltaRt and deltaRzDIN, respectively, and 54.8, 516.6 and 258.2 nm after 1-min immersion in 1.0% citric acid. Changes of the surface roughness were dependent on the exposure time and concentration of acidic solution. Pellicle layer significantly reduced the extent of erosive destruction, which was additionally documented on SEM-micrographs. Residual pellicle-like structures were detected after 5 min of immersion in 0.1% citric acid. However, there were no significant differences in pellicle-covered and non-covered enamel slabs measured profilometrically for 1.0% citric acid with 10 min and 30 min exposure time. The findings confirm the property of pellicle layer to resist against erosive influence of organic acids, which is, however, limited by duration of acidic treatment and concentration of erosive agent.


Gd-HPDO3A, a complex to obtain high-phasing-power heavy-atom derivatives for SAD and MAD experiments: results with tetragonal hen egg-white lysozyme.
A neutral gadolinium complex, Gd-HPDO3A, is shown to be a good candidate to use to obtain heavy-atom derivatives and solve macromolecular structures using anomalous dispersion. Tetragonal crystals of a gadolinium derivative of hen egg-white lysozyme were obtained by co-crystallization using different concentrations of the complex. Diffraction data from three derivative crystals (100, 50 and 10 mM) were collected to a resolution of 1.7 A using Cu Kalpha radiation from a rotating anode. Two strong binding sites of the gadolinium complex to the protein were located from the gadolinium anomalous signal in both the 100 and 50 mM derivatives. A single site is occupied in the 10 mM derivative. Phasing using the anomalous signal at a single wavelength (SAD method) leads to an electron-density map of high quality. The structure of the 100 mM derivative has been refined. Two molecules of the gadolinium complex are close together. Both molecules are located close to tryptophan residues. Four chloride ions were found. The exceptional quality of the SAD electron-density map, only enhanced by solvent flattening, suggests that single-wavelength anomalous scattering with the Gd-HPDO3A complex may be sufficient to solve protein structures of high molecular weight by synchrotron-radiation experiments, if not by laboratory experiments.


[Obtaining iodinated luteinizing hormone while preserving its biological properties].
The author describes a method of obtaining biologically-active 125I-labeled luteinzing hormone in iodation with chloramine T. The hormone and chloramine T concentration ratio of 1:2, and the reaction time of 20 sec was used. LH-125I with the specific activity of 27--30 microCi/microgram was bound with the receptor strictly specifically.


Bioaccumulation and biotransformation of polycyclic aromatic hydrocarbons during sediment tests with oligochaetes (Lumbriculus variegatus).
In some kinetic studies with aquatic invertebrates, the bioaccumulation of polyaromatic hydrocarbons (PAHs) has been observed to peak at the beginning of the test. This has been explained by the depletion of PAHs from pore water due to limited desorption during the bioaccumulation test or, alternatively, by the activation of biotransformation mechanisms in the organisms. In the present study, we exposed the aquatic oligochaetes, Lumbriculus variegatus, to creosote oil-contaminated sediments to examine the bioaccumulation of PAHs and to clarify the importance of contaminant depletion and biotransformation for it. The contaminant depletion was studied by replanting test organisms into fresh, nondepleted test sediments at 3-d intervals over 12 d and by comparing the resulting body burdens to those of the organisms that were not replanted. The biotransformation capability of L. variegatus was assessed by following the concentration of 1-hydroxypyrene (1-HP), a phase I metabolite of pyrene, in oligochaete tissue during a 15-d test. We observed that the bioaccumulation of most PAHs indeed peaked at the beginning of the test. The concentrations in the replanted organisms were only 1.5 to 2 times higher than in nonreplanted organisms during the first 9 d of the test and, by day 12, no differences were detected. 1-Hydroxypyrene was detected in oligochaete tissue throughout the exposures, and concentrations decreased over time. However, the proportion of 1-HP to pyrene increased linearly during the test. These results indicated that the depletion of contaminants has only a minor effect on their bioaccumulation in oligochaetes and that the cause for the observed bioaccumulation curve shape is rapid elimination of the contaminants and, possibly to some degree, their metabolites.


A Unique Clinicopathological Manifestation of Fungal Infection: A Case Series of Deep Dermatophytosis in Immunosuppressed Patients.
Dermatophytes are the most common cause of superficial fungal infections in humans. Deep dermatophytosis, however, is rare, described to date only in isolated case reports, usually in the setting of systemic immunosuppression. To present the 15-year experience of a tertiary dermato-mycology clinic with the diagnosis and treatment of deep dermatophytosis. Patients were identified by database search. Clinical, mycological, histological, and treatment data were collected from the medical files. Ten patients were identified: nine after solid-organ transplantation and one undergoing chemotherapy, all diagnosed within 3 years after beginning immunosuppression (average 7.5 months). The infective agent in nine cases was Trichophyton rubrum. All patients presented with concurrent superficial fungal infections. Complete resolution was noted in response to systemic antifungal agents. There was no histological evidence of hair-follicle involvement. The limitations of the study were the retrospective design and the small cohort size. This case-series study suggests that deep dermatophytosis is a separate entity, distinct from Majocchi's granuloma. It occurs only in immunocompromised patients and is characterized by discrete nodules, an indolent course, the absence of follicular invasion, and proximity to a superficial dermatophyte infection. Systemic antifungal treatment leads to complete resolution. The urgent need for the treatment of superficial fungal infections in immunocompromised patients is emphasized.


Combined corneal collagen cross-linking and posterior chamber toric implantable collamer lens implantation for keratoconus.
A 29-year-old woman with keratoconus underwent posterior chamber toric implantable Collamer lens (Visian ICL; STAAR Surgical, Monrovia, CA) implantation 12 months after corneal collagen cross-linking with riboflavin and ultraviolet radiation-A. No intraoperative or postoperative complications were observed. Three months postoperatively, uncorrected distance visual acuity improved from counting fingers to 20/40 and corrected distance visual acuity improved from 20/100 to 20/30. Combined cross-linking treatment and posterior chamber toric phakic Collamer lens implantation in a two-step procedure seems to be an effective method for correcting keratoconus in patients with high myopia and astigmatism. Despite the encouraging results, longer follow-up is necessary to evaluate the outcomes of this approach.


Perceived location of two-dimensional patterns.
The study attempted to test the possibility that the center of gravity of two-dimensional patterns is the cue used by a human observer for their localization. Four experiments were carried out. The first, using a matching procedure, required the localization of the center of irregular dot patterns, contour and filled polygons which varied in size and orientation. In the other three experiments the subjects had to point to briefly exposed dot patterns in which overall shape (convex and concave in Expts 2 and 3) and dot density (Expt 4) were manipulated. The performance of these direct localization tasks was found to be as accurate as the performance in previous studies of indirect localization or regular patterns. The results consistently supported the claim that information about position of the center of gravity is used for the localization of visual objects.


Biological tests on an experimental glass ionomer (silicopolyacrylate) cement.
The biological compatibility of an experimental glass ionomer (silicopolyacrylate) cement has been assessed in in vitro (cell culture) and in vivo (monkey teeth) tests. For comparison a conventional silicate cement (Super Syntrex) was included in the in vivo experiments. In vitro, the glass ionomer cement was toxic when freshly prepared. The toxicity decreased, however, with increasing setting time. In experiments with prolonged cell-material contact time with specimens which had set for 24 h, the material appeared to be non-toxic. The in vivo experiments confirmed previous reports concerning unfavourable pulp reaction caused by silicate cement, while the glass ionomer cement caused mainly a mild pulp reaction after 8 days of observation. Unfortunately no longer-term observations were obtained since the material had been washed out of the cavities within 36 days.


Single-ensemble nonequilibrium path-sampling estimates of free energy differences.
We introduce a straightforward, single-ensemble, path sampling approach to calculate free energy differences based on Jarzynski's relation. For a two-dimensional "toy" test system, the new (minimally optimized) method performs roughly one hundred times faster than either optimized "traditional" Jarzynski calculations or conventional thermodynamic integration. The simplicity of the underlying formalism suggests the approach will find broad applicability in molecular systems.


Tick paralysis.
The one tick-borne disease that rarely comes under the auspices of the infectious disease specialist is not caused by an infectious agent, but is tick paralysis. This condition is caused by tick bite and typically presents as a flaccid ascending paralysis. This article discusses this entity partly because of completeness, but also because tick paralysis, or tick toxicosis as it is sometimes called, is worth the infectious disease consultant's consideration. The differential diagnosis includes entities that are infectious or caused by toxins of infectious agents, such as epidural abscess, some causes of transverse myelitis, and botulism. Lastly, in an era of antibiotic toxicity, multidrug-resistant bacteria, antigen-switching viruses, and complex antibiotic regimens, the cure for tick paralysis-removing the tick-is as simple as it is gratifying.


Effects of high frequency repetitive transcranial magnetic stimulation on P(300) event-related potentials.
Auditory event-related potentials (P(300)-ERPs) were analyzed before and after repetitive transcranial magnetic stimulation (rTMS). Two rTMS trains (10 Hz, 3 s, 100% motor threshold and 5 min interval) were delivered over the left frontal area in healthy subjects. P(300)-ERPs were recorded at 14 electrode sites on the scalp using a typical oddball paradigm before and after rTMS. The latencies and amplitudes of N(100), P(200), N(200) and P(300) were measured and compared. The directed coherence (DCOH) was estimated to demonstrate information flow between different cortical areas. rTMS significantly influenced P(300)-ERPs. The effects differed on the different components (P<0.001). The latency of P(300) significantly increased after stimulation, which was more obvious in the frontal and central areas. The changes in P(300) amplitude were not significant (P>0.05). The DCOH from the frontal, central, parietal and occipital areas to the temporal area was significantly higher than the DCOH from the temporal area to the former 4 areas (P<0.01). rTMS with the present parameters can affect P(300)-ERPs, leading to a delayed P(300) component and changes in information connections around the stimulated site. Our data suggest that rTMS may postpone neuronal activities related to cognitive processing.


Solid-phase synthesis of kojic acid-tripeptides and their tyrosinase inhibitory activity, storage stability, and toxicity.
A small library of kojic acid-tripeptides (Ko-X1X2X3) was prepared by solid-phase parallel synthesis and assayed to evaluate their tyrosinase inhibitory activity. Most of the kojic acid-tripeptides showed better activities than kojic acid. Kojic acid-FWY was the best compound, and it exhibited 100-fold tyrosinase inhibitory activity compared with kojic acid. In addition, their storage stabilities were approximately 15 times higher and their toxicity was lower than that of kojic acid.


"Silent" MRI with soft gradient pulses.
A method to reduce the acoustic noise generated by gradient systems in magnetic resonance imaging (MRI) is proposed based on the linear response theory. Since the acoustic frequency response function of typical gradient coils is low in the range below 200 Hz, the noise level can be significantly reduced by using gradient pulse sequences whose spectra are limited to this frequency range. Such "soft," i.e., band-limited, pulse shapes can be designed using sinusoidal ramps individually adjusted to available delays. "Silent" versions of three basic MRI sequences [gradient-echo (GE), spin-echo (SE), and rapid acquisition with relaxation enhancement (RARE)] were programmed on 2 and 3 T whole-body scanners. High-quality images could be acquired at noise levels as low as 40 dBA (GE and SE) and 60 dBA (RARE).


Identifying best methods for routine ELISA detection of microcystin in seafood.
Ingestion of water contaminated with the cyanotoxin, microcystin (MC), can pose serious health risks to humans. MC is also known to accumulate in seafood; however, this exposure pathway is much less understood. A fundamental element of this uncertainty is related to analytical difficulties. Commercially available enzyme-linked immunosorbent assays (ELISAs) offer one of the best options for routine MC detection, but methods of detecting MC in tissue are far from standardized. We spiked freshwater finfish and marine mussel tissues with MC, then compared recovery rates using four different preparation protocols and two ELISA types (polyclonal anti-MC-ADDA/direct monoclonal (DM)). Preparation protocol, type of ELISA, and seafood tissue variety significantly affected MC detection. This is the first known study to use DM ELISA for tissue analyses, and our findings demonstrate that DM ELISA combined with a short solvent extraction results in fewer false positives than other commonly used methods. This method can be used for rapid and reliable MC detection in seafood.


Molecular events underlying skeletal muscle atrophy and the development of effective countermeasures.
Skeletal muscle adapts to loading; atrophying when exposed to unloading on Earth or in spaceflight. Significant atrophy (decreases in muscle fiber cross-section of 11-24%) in humans has been noted after only 5 days in space. Since muscle strength is determined both by muscle cross-section and synchronization of motor unit recruitment, a loss in muscle size weakens astronauts, which would increase risks to their safety if an emergency required maximal muscle force. Numerous countermeasures have been tested to prevent atrophy. Resistant exercise together with growth hormone and IGF-I are effective countermeasures to unloading as most atrophy is prevented in animal models. The loss of muscle protein is due to an early decrease in protein synthesis rate and a later increase in protein degradation. The initial decrease in protein synthesis is a result of decreased protein translation, caused by a prolongation in the elongation rate. A decrease in HSP70 by a sight increase in ATP may be the factors prolonging elongation rate. Increases in the activities of proteolytic enzymes and in ubiquitin contribute to the increased protein degradation rate in unloaded muscle. Numerous mRNA concentrations have been shown to be altered in unloaded muscles. Decreases in mRNAs for contractile proteins usually occur after the initial fall in protein synthesis rates. Much additional research is needed to determine the mechanism by which muscle senses the absence of gravity with an adaptive atrophy. The development of effective countermeasures to unloading atrophy will require more research.


Phase II trial of adjuvant chemotherapy with bi-weekly carboplatin plus paclitaxel in patients with completely resected non-small cell lung cancer.
Adjuvant chemotherapy improves the prognosis of patients with non-small cell lung cancer (NSCLC) after a complete resection despite unacceptable toxicity and low compliance. A total of 67 patients were enrolled in a multi-institutional study. The patients received chemotherapy with carboplatin (CBDCA) area under the curve of 3 and paclitaxel (PTX) 90 mg/m(2) every 2 weeks for six cycles after surgery. Fifty patients (74.6%) completed all cycles of therapy. The presence of grade 3 and 4 toxicities of neutropenia were 13.4, and 3.0%, respectively. Non-haematological adverse effects were infrequent and no treatment-related death was registered. The estimated disease-free survival and overall survival at 2 years were 89.0% and 88.8%, respectively. A bi-weekly schedule of CBDCA and PTX as adjuvant chemotherapy showed an acceptable toxicity and favourable feasibility in Japanese NSCLC patients after complete tumor resection. Consequently, it is desirable to validate this regimen in a future randomized clinical trial.


Ultrastructural and tissue-culture studies on the role of fibronectin, collagen and glycosaminoglycans in the migration of neural crest cells in the fowl embryo.
The initial migration of neural crest (NC) cells into cell-free space was studied by transmission electron microscopy at trunk levels of fowl embryos, some of which were fixed in the presence of ruthenium red. Migrating NC cells occurred in zones which contained fewer ruthenium-red stained 15-40nm diameter granules than other regions. The ruthenium-red stained granules were linked by similarly stained thin (greater than 3nm diameter) microfibrils. The granules resemble proteoglycan and the microfibrils may be hyaluronate. NC cells contacted thicker (greater than 10 nm diameter) fibrils and interstitial bodies, which did not require ruthenium red for visualization. Cytoplasmic microfilaments were sometimes aligned at the point of contact with the extracellular fibrils, which may be fibronectin and collagen. Phase-contrast time-lapse videotaping and scanning electron microscopy showed that NC cells of the fowl embryo in vitro migrated earlier and more extensively on glass coated with fibronectin-rich fibrous material and adsorbed fibronectin molecules than on glass coated with collagen type I (fibres and adsorbed molecules). NC cells became completely enmeshed in fibronectin-rich fibres, but generally remained on the surface of collagen-fibre gels. When given a choice, NC cells strongly preferred fibronectin coatings to plain glass, and plain glass to dried collagen gels. NC cells showed a slight preference for plain glass over glass to which collagen was adsorbed. Addition to the culture medium of hyaluronate (initial conc. 20 mg/ml), chondroitin (5 mg/ml) and fully sulphated chondroitin sulphate and dermatan sulphate (up to 10 mg/ml) did not drastically alter NC cell migration on fibronectin-rich fibrous substrates.


The role of 2'-5' oligoadenylate-activated ribonuclease L in apoptosis.
Apoptosis of viral infected cells appears to be one defense strategy to limit viral infection. Interferon can also confer viral resistance by the induction of the 2-5A system comprised of 2'-5' oligoadenylate synthetase (OAS), and RNase L. Since rRNA is degraded upon activation of RNase L and during apoptosis and since both of these processes serve antiviral functions, we examined the role RNase L may play in cell death. Inhibition of RNase L activity, by transfection with a dominant negative mutant, blocked staurosporine-induced apoptosis of NIH3T3 cells and SV40-transformed BALB/c cells. In addition, K562 cell lines expressing inactive RNase L were more resistant to apoptosis induced by decreased glutathione levels. Hydrogen peroxide-induced death of NIH3T3 cells did not occur by apoptosis and was not dependent upon active RNAse L. Apoptosis regulatory proteins of the Bcl-2 family did not exhibit altered expression levels in the absence of RNase L activity. RNase L is required for certain pathways of cell death and may help mediate viral-induced apoptosis.


Hierarchical estimation of a dense deformation field for 3-D robust registration.
A new method for medical image registration is formulated as a minimization problem involving robust estimators. We propose an efficient hierarchical optimization framework which is both multiresolution and multigrid. An anatomical segmentation of the cortex is introduced in the adaptive partitioning of the volume on which the multigrid minimization is based. This allows to limit the estimation to the areas of interest, to accelerate the algorithm, and to refine the estimation in specified areas. At each stage of the hierarchical estimation, we refine current estimate by seeking a piecewise affine model for the incremental deformation field. The performance of this method is numerically evaluated on simulated data and its benefits and robustness are shown on a database of 18 magnetic resonance imaging scans of the head.


Genetic diversity of two haploid markers in the Udegey population from southeastern Siberia.
The Udegeys are a small ethnic group who live along the tributaries of the Amur River Basin of southeastern Siberia in Russia. They are thought to speak a language belonging to a subdivision of the Tungusic-Manchu branch of the Altaic family. To understand the genetic features and genetic history of the Udegeys, we analyzed two haploid markers, mitochondrial DNA (mtDNA), and Y-chromosomal variation, in 51 individuals (including 21 males) from the Udegey population. In general, the Udegeys' mtDNA profiles revealed similarities to Siberians and other northeastern Asian populations, although a moderate European contribution was also detected. Interestingly, pairwise values of F(ST) and the MDS plots based on the mtDNA variation showed that the Orok and Nivkh inhabiting the very same region of the Udegey were significantly different from the Udegey, implying that they may have been isolated and undergone substantial genetic drift. The Udegeys were characterized by a high frequency (66.7%) of Y chromosome haplogroup C, indicating a close genetic relationship with Mongolians and Siberians. On the paternal side, however, very little admixture was observed between the Udegeys and Europeans. Thus, the combined haploid genetic markers of both mtDNA and the Y chromosome imply that the Udegeys are overall closest to Siberians and northeast Asians of the Altaic linguistic family, with a minor maternal contribution from the European part of the continent.


Nasa astronauts, prosthetics and the manned space program.
The author has collaborated with NASA astronauts, scientists and engineers since 2006. Manned deep space missions, beyond the Moons orbit, are being planned in this post-Shuttle era. The spacecraft required for longer flights will have relatively restricted crew interior volume. To decrease the negative impact of these tight quarters, the author has proposed recruiting smaller astronauts (abbreviated SAs), persons about one-half the height of current near-Earth crewmembers. This includes achondroplastic dwarfs, lower extremity amputees and persons with certain height-reducing birth defects such as phocomelia. To overcome issues of physical competence, strength and mobility of SAs, the author describes using advanced cybernetic prostheses for those with limb amputations or deformities, and motorized exoskeletons for the others. Muscle and bone-sparing space exercise programs for SAs should be simpler. For example, a motorized exoskeleton used for routine duties in space would also provide both resistance workouts and passive range of motion conditioning for the astronauts, even while resting. Complex personalized artificial intelligence functions may be added. These initial suggestions previously presented to NASA offer a starting point for deep space manned missions to the asteroid belt, Mars and beyond.


Ultrastructure of avian gastrin cell granules.
Gastrin cells in the pigeon, quail, gull and kite were identified at ultrastructural level by immunocytochemistry, using the consecutive semithin/ultrathin section technique. In contrast to the gastrin granules known in mammals, the avian gastrin granules generally contained a consistent dense core accompanied by a clear halo between the core and limiting membrane. The mean diameters of gastrin granules in the pigeon, quail, gull and kite were 211 +/- 37, 247 +/- 45, 331 +/- 61 and 353 +/- 73 nm respectively. The carnivorous birds seemed to possess larger gastrin granules than the grain-eating birds. The species differences in the size and fine structures of the gastrin granules were discussed.


Electrophysiological evidence for A10 dopamine autoreceptor subsensitivity following chronic D-amphetamine treatment.
Extracellular single unit recording techniques were used to determine whether chronic treatment with D-amphetamine (AMP) causes a subsensitivity of dopamine (DA) autoreceptors on A10 DA neurons in the rat ventral tegmental area. Either once daily or twice daily intraperitoneal (i.p.) administration of 5.0 mg/kg AMP for 1 week significantly reduced the ability of intravenous (i.v.) AMP and apomorphine (APO) to suppress the firing of A10 DA neurons when tested 24-32 h after the final administration of i.p. AMP. For both of these treatment regimens, the dose-response curves for AMP and APO induced suppression were shifted approximately 4-fold to the right of control. Following an 8 day abstinence period, only the rats that received twice daily AMP injections exhibited subsensitivity to i.v. AMP and APO; the degree of subsensitivity was reduced by 50% as compared to that observed 24-32 h post-treatment. These results were not due to acute tolerance phenomena since a single i.p. injection of AMP 24-32 h before testing failed to alter sensitivity to i.v. AMP and APO. Rather, the results indicate that chronic AMP treatment reduces the sensitivity of A10 DA neurons to DA agonists. DA autoreceptor subsensitivity was demonstrated further by the finding that the ability of microiontophoretically applied DA to suppress A10 DA neuronal activity was markedly reduced (5.8-fold shift of the dose-response curve) by chronic AMP treatment (2 X 5 mg/kg/day). In contrast, there was no alteration in the ability of iontophoretic gamma-amino butyric acid (GABA) to suppress A10 DA activity in chronic AMP rats. Chronic AMP-treatment also increased the number of spontaneously active A10 DA neurons as well as their basal firing rate. It is suggested that the ability of chronic AMP treatment to decrease the auto-regulatory ability of A10 DA neurons may be related to the phenomena of behavioral sensitization and AMP psychosis.


Exposure-Efficacy Analyses of Ombitasvir, Paritaprevir/Ritonavir with Dasabuvir ± Ribavirin in HCV Genotype 1-Infected Patients.
The three-direct-acting antiviral (DAA) combination regimen of ombitasvir, paritaprevir (coadministered with ritonavir [paritaprevir/ritonavir], and dasabuvir (the 3D regimen) ± ribavirin for treatment of HCV genotype 1-infected patients demonstrated efficacy and safety in Phase II and Phase III clinical trials. The relationships between the steady-state exposure (area under the concentration-time curve at steady state and trough concentration at steady state) of the three DAAs and ribavirin with sustained virologic response at 12 weeks after treatment (SVR12) following administration of the 3D regimen in six Phase II/III studies were examined. HCV non-cirrhotic genotype 1-infected adult male and female patients (N = 1690) enrolled in the one Phase II study or one of the five Phase III studies were included for graphical analysis. HCV subgenotype 1a-infected patients who received the 3D regimen with ribavirin (approved regimen for that patient population) (N = 615) from the same studies were included in the multivariate logistic regression exposure-response analysis. Graphical analysis suggested a shallow trend between exposure and % SVR12 for paritaprevir, ombitasvir, and ribavirin exposure but not for dasabuvir exposure. After adjusting for covariate effects, the exposure-response logistic-regression analysis indicated that ombitasvir exposure was the single significant predictor, demonstrating a 1 % change in SVR12 with up to 25 % change in ombitasvir exposure at steady state. The results of these analyses indicate that the doses selected for the 3D regimen were optimal, achieving high SVR12 rates across the range of exposures observed in the Phase III studies.


Enzymatic modification of the surface carbohydrates of Friend erythroleukemic cells.
Surface carbohydrates of Friend erythroleukemic-cells were modified by treatment with the exoglycosidases, alpha-galactosidase, beta-galactosidase, and neuraminidase without affecting cell growth and viability either in the presence of absence of 1.8% DMSO as inducer. When cells were incubated with a combination of alpha-galactosidase and neuraminidase and then induced, they showed an increased rate of differentiation as measured by the formation of benzidine-positive cells. These enzymes used singly, or beta-galactosidase treatment alone, or in combination with neuraminidase, did not change the rate of differentiation. Cell-surface labeling and electrophoretic separation of the glycoconjugates revealed that two regions of approximate molecular weights of 195,000 and 185,000 were neuraminidase-sensitive and one other of molecular weight of about 75,000 was sensitive to alpha-galactosidase. Both untreated and the combined alpha-galactosidase, neuraminidase-modified cells exhibited the same rate of uptake of carbon-14 DMSO, ruling out the possibility that the observed increased rate of differentiation was due to faster penetration of DMSO into enzyme-treated cells. On the other hand, the decrease in the rate of uptake of rubidium-86, an analogue of K+, by treated-induced cells was significantly enhanced over that observed with untreated-induced cells, suggesting that alpha-galactosidase plus neuraminidase modification of the cell surface was affecting at least one of the early events occurring in the Friend erythroleukemic cell differentiation program.


Cloning and characterization of the acyl-coenzyme A: 6-aminopenicillanic-acid-acyltransferase gene of Penicillium chrysogenum.
A gene, aat, encoding acyl-CoA: 6-aminopenicillanic acid acyltransferase (AAT), the last enzyme of the penicillin (Pn) biosynthetic pathway, has been cloned from the genome of Penicillium chrysogenum AS-P-78. The gene contains three introns in the 5'-region and encodes a protein of 357 amino acids with an Mr of 39,943. It complements mutants of P. chrysogenum deficient in AAT activity. The aat gene is expressed as a 1.15-kb transcript and the encoded protein appears to be processed post-translationally into two nonidentical polypeptides of 102 and 255 aa, with Mrs of 11,498 and 28,461, respectively. Three proteins of 40, 11, and 29 kDa (the last one corresponding to the previously purified AAT), were identified in extracts of P. chrysogenum. The aa sequence of the N-terminal end of the 11-kDa polypeptide matched the nucleotide (nt) sequence of the 5'-region of aat. The N-terminal end of the 29-kDa polypeptide corresponded to the sequence beginning at nt position 916 of the sequenced DNA fragment (nt 441 of aat gene). The aat gene of P. chrysogenum resembles the genes encoding Pn acylases of Escherichia coli, Proteus rettgeri and Pseudomonas sp., all of which encode two nonidentical subunits derived from a common precursor, encoded by a single open reading frame.


Home care nurses' provision of support to families of the elderly at the end of life.
In this article we describe our study of assistance for family decisions and caregiving by Japanese home care nurses to families of elderly relatives at the end of life. The participants were 31 nurses who had been evaluated as providing good end-of-life care. We carried out semistructured interviews concerning the practice of family support in two cases (cancer and noncancer). We conducted a qualitative analysis using the constant comparative approach and derived several categories inductively. Home care nurses are responsible for (a) estimating the possibility of dying at home, (b) visualizing what is coming and what can be done, (c) proposing where and how the family can say goodbye, (d) building family consensus, (e) coordinating resources, and (f) offering psychological support for end-of-life care. End-of-life family care by home care nurses is a process in which multiple components of care are provided with changing content as death approaches.


Coexistence of renin and angiotensin II in epitheloid cell secretory granules of rat kidney.
The distribution of renin and angiotensin II (ANG II) in juxtaglomerular epitheloid cells of control and adrenalectomized rats was studied, using specific antisera and the protein A-gold technique in Lowicryl- and glycol methacrylate-embedded tissue. The matrix of virtually all mature secretory granules of epitheloid cells contains not only renin, but also ANG II. On adrenalectomy, the concentration of both renin and On adrenalectomy, the concentration of both renin and ANG II in the granule internum increases markedly, as indicated by the density of the immunolabel. Given the coexistence of renin and ANG II in the granule matrix, it is quite probable that, with each secretory event, a certain amount of ANG II is released together with renin. Further experiments will have to show if this amount of ANG II cosecreted with renin is sufficient to elicit immediate local intrarenal actions. ANG I, as well as angiotensinogen and converting enzyme, were not found in epitheloid cells. It is therefore inferred that ANG II is not generated intracellularly, but within the extracellular space and subsequently taken up by pinocytosis and incorporated into the secretory granules of epitheloid cells.


Gene expression profiling of seizure disorders.
Seizure disorders affect a significant percentage of the population, and researchers worldwide continue to work toward a better understanding of what initiates, propagates, and results from aberrant and excessive neuronal excitation. During the past two decades, one aspect of this research effort has been to describe the effects of seizure activity upon neuronal gene expression, with hopes of identifying the molecular mechanisms that underlie subsequent changes in cell function and survival. Here we review this body of work from the perspective of how these gene profiling efforts have evolved, starting with one-by-one analyses of specific gene targets to the more recent use of DNA microarrays to survey literally thousands of genes simultaneously. With regard to the latter, we present some of our own work that suggests that molecular mechanisms contributing to normal brain development are reiterated during seizure-induced network reorganization.


Which opioid receptor mechanism modulates feeding?
There is substantial evidence for the role of endogenous opioid peptides in the regulation of appetite. This communication examines the possible opioid peptide mechanism(s) which are involved in appetite regulation. In the rat, activation of both the dynorphin-kappa opioid receptor and the beta-endorphin-epsilon opioid receptor appear to enhance feeding, most probably acting in different areas of the central nervous system. It also appears that rats may have a mu anorectic system. Too few studies have been undertaken to define whether the delta or sigma receptor systems are also involved in feeding responses. It is becoming apparent that a great deal of species diversity exists in the feeding responses to opiates, making it difficult to extrapolate the results obtained in rats to other species. In humans, studies with naloxone suggest an opioid sensitive feeding system which possibly is specifically involved in the regulation of carbohydrate uptake. In addition, we report here preliminary data suggesting the presence of a mu anorectic system in humans. Thus, analogous to the findings for the role of opioid receptors in analgesia, it appears that multiple opioid receptors may be involved in appetite regulation, each receptor relating to a different aspect of feeding.


Validation of an MPC Polymer Coating to Attenuate Surface-Induced Crosstalk between the Complement and Coagulation Systems in Whole Blood in In Vitro and In Vivo Models.
Artificial surfaces that come into contact with blood induce an immediate activation of the cascade systems of the blood, leading to a thrombotic and/or inflammatory response that can eventually cause damage to the biomaterial or the patient, or to both. Heparin coating has been used to improve hemocompatibility, and another approach is 2-methacryloyloxyethyl phosphorylcholine (MPC)-based polymer coatings. Here, the aim is to evaluate the hemocompatibility of MPC polymer coating by studying the interactions with coagulation and complement systems using human blood in vitro model and pig in vivo model. The stability of the coatings is investigated in vitro and MPC polymer-coated catheters are tested in vivo by insertion into the external jugular vein of pigs to monitor the catheters' antithrombotic properties. There is no significant activation of platelets or of the coagulation and complement systems in the MPC polymer-coated one, which was superior in hemocompatibility to non-coated matrix surfaces. The protective effect of the MPC polymer coat does not decline after incubation in human plasma for up to 2 weeks. With MPC polymer-coated catheters, it is possible to easily draw blood from pig for 4 days in contrast to the case for non-coated catheters, in which substantial clotting is seen.


A novel artificial prosthetic replacement for the proximal interphalangeal joint of the hand--from concept to prototype.
This article describes the development of a proximal interphalangeal (PIP) joint prosthesis based on the principles of replicating anatomical surface components, the use of macrolocking intramedullary stem and the use of a cobalt-chrome alloy material. The design features are intended to obtain an optimal range of motion while retaining stability and longevity. The final prototype, for which a patent has been filed, is described.


[A method of determination of cholesterol contents on the surface of the human skin and prospects of its use. II. Psoriasis].
A noninvasive method for cholesterol extraction from the surface layers of the epidermis is suggested. Levels of cholesterol and its fractions were measured in extracts obtained from the foci of psoriasis and apparently intact skin of 105 patients with psoriasis. Cholesterol levels were found noticeably increased, mainly at the expense of its free fraction in involved foci and in apparently intact skin. A conclusion is made on the good informative value of the suggested method.


Design, synthesis and preliminary antiproliferative activity studies of new diheteroaryl thioether derivatives.
A series of structurally new diheteroaryl thioether analogs was designed, prepared and screened toward MGC-803, MKN-45, EC-109 and H1650. Most of the target compounds displayed moderate to potent antiproliferative activities. Among them, compound 5 showed the best antiproliferative activity against the tested cell lines with the half maximal inhibitory concentration (IC50) values below 10μM. In addition, flow cytometry analysis showed that compound 5 increased Bax expression, down-regulated expression of Bcl-2, cleaved caspases-3/9, finally inducing apoptosis of MKN-45 cells as well asarrested the cell cycle at G2/M phase. This study suggests that the diheteroaryl thioethers are a class of emerging chemotypes for developing antitumor agents or biological probes, and compound 5 could serve as a good starting point to design new apoptosis inducers.


Effect of point mutations on the ultrafast photo-isomerization of Anabaena sensory rhodopsin.
Anabaena sensory rhodopsin (ASR) is a particular microbial retinal protein for which light-adaptation leads to the ability to bind both the all-trans, 15-anti (AT) and the 13-cis, 15-syn (13C) isomers of the protonated Schiff base of retinal (PSBR). In the context of obtaining insight into the mechanisms by which retinal proteins catalyse the PSBR photo-isomerization reaction, ASR is a model system allowing to study, within the same protein, the protein-PSBR interactions for two different PSBR conformers at the same time. A detailed analysis of the vibrational spectra of AT and 13C, and their photo-products in wild-type ASR obtained through femtosecond (pump-) four-wave-mixing is reported for the first time, and compared to bacterio- and channelrhodopsin. As part of an extensive study of ASR mutants with blue-shifted absorption spectra, we present here a detailed computational analysis of the origin of the mutation-induced blue-shift of the absorption spectra, and identify electrostatic interactions as dominating steric effects that would entail a red-shift. The excited state lifetimes and isomerization reaction times (IRT) for the three mutants V112N, W76F, and L83Q are studied experimentally by femtosecond broadband transient absorption spectroscopy. Interestingly, in all three mutants, isomerization is accelerated for AT with respect to wild-type ASR, and this the more, the shorter the wavelength of maximum absorption. On the contrary, the 13C photo-reaction is slightly slowed down, leading to an inversion of the ESLs of AT and 13C, with respect to wt-ASR, in the blue-most absorbing mutant L83Q. Possible mechanisms for these mutation effects, and their steric and electrostatic origins are discussed.


Clinical significance of preoperative serum albumin level for prognosis in surgically resected patients with non-small cell lung cancer: Comparative study of normal lung, emphysema, and pulmonary fibrosis.
This study was performed to clarify whether preoperative serum albumin level is related to the prognosis of non-small cell lung cancer patients undergoing surgical resection, and the relationships between serum albumin level and clinicopathological characteristics of lung cancer patients with emphysema or pulmonary fibrosis. We retrospectively evaluated 556 patients that underwent surgical resection for non-small cell lung cancer. The correlation between preoperative serum albumin level and survival was evaluated. Patients were divided into three groups according to the findings on chest high-resolution computed tomography (normal lung, emphysema, and pulmonary fibrosis), and the relationships between serum albumin level and clinicopathological characteristics, including prognosis, were evaluated. The cut-off value of serum albumin level was set at 4.2g/dL. Patients with low albumin levels (albumin <4.2) had significantly poorer prognosis than those with high albumin levels (albumin ≥4.2) with regard to both overall survival and recurrence-free survival. Serum albumin levels in the emphysema group (n=48) and pulmonary fibrosis group (n=45) were significantly lower than that in the normal lung group (n=463) (p=0.009 and <0.001, respectively). Low serum albumin level was a risk factor in normal lung and pulmonary fibrosis groups, but not in the emphysema group. Preoperative serum albumin level was an important prognostic factor for overall survival and recurrence-free survival in patients with resected non-small cell lung cancer. Divided into normal lung, emphysema, and pulmonary fibrosis groups, serum albumin level showed no influence only in patients in the emphysema group.


The glucose-6-phosphatase system in human development.
The classical role of glucose-6-phosphatase in liver and kidney is the production of glucose for release into blood. In liver, glucose-6-phosphatase catalyses the terminal step of glycogenolysis and gluconeogenesis. Abnormally low hepatic glucose-6-phosphatase activity is found in human genetic deficiencies i.e. glycogen storage disease type I and in cases of developmental delay, found predominantly in preterm infants. In contrast, abnormally high liver glucose-6-phosphatase occurs in poorly controlled or untreated diabetes mellitus. Hepatic glucose-6-phosphatase is an integral endoplasmic reticulum (and nuclear membrane) protein and it is part of a multicomponent system. Its active site is situated inside the lumen of the endoplasmic reticulum and transport proteins are needed to allow its substrates glucose-6-phosphate (and pyrophosphate) and its products phosphate and glucose to cross the endoplasmic reticulum membrane. In addition, a calcium binding protein is also associated with the glucose-6-phosphatase enzyme. Immunohistochemical studies, in combination with image analysis, have shown that glucose-6-phosphatase is present in liver and kidney and also in specific cell types in a variety of human tissues, for example Leydig cells in the testis and some astrocytes in the brain. Where practicable, enzymatic analysis, direct transport assays and/or immunological detection of the endoplasmic reticulum glucose and phosphate transport proteins have been used to demonstrate the presence and activity of the whole glucose-6-phosphatase system. The distribution of the human glucose-6-phosphatase system changes dramatically during development with a different spatial and temporal pattern in each tissue. The most unexpected localization was in circulating, predominantly nucleated, embryonic and early fetal red blood cells.


No effect of waxed dental floss on enamel fluoride uptake or on enamel dissolution changes.
Enamel specimens were flossed in vitro with "waxed" dental floss and then treated with water, Act, Meridol, or Elmex Gel to evaluate the effect of "waxed" floss on the enamel fluoride uptake and on the enamel dissolution rate. The flossed and treated enamel was etched 4 times successively in 2N HCl for 5, 10, 15 and another 15 s. The concentrations of fluoride, calcium and phosphorus were assayed in the etching solutions. The enamel fluoride concentration was significantly increased in only the first layer of the Elmex Gel group compared to the other 3 groups. Respective etching depths were similar for the water, Act and Elmex Gel groups. However, the first layer in the Meridol group was less etched than that in the 3 other groups. This study showed that "waxed" dental floss did not compromise the fluoride uptake by surface enamel after treatment with a concentrated acidic topical fluoridating agent. Moreover, previously reported results showing that Meridol solution containing SnF2 and amine fluoride 297 significantly decreased the in vitro dissolution rate of enamel were confirmed by the present study. Even though the enamel was first treated with "waxed" dental floss, the SnF2/amine fluoride 297 still produced a decreased enamel dissolution rate.


Activities of N-Myc in the developing limb link control of skeletal size with digit separation.
The developing limb serves as a paradigm for studying pattern formation and morphogenetic cell death. Here, we show that conditional deletion of N-Myc (Mycn) in the developing mouse limb leads to uniformly small skeletal elements and profound soft-tissue syndactyly. The small skeletal elements are associated with decreased proliferation of limb bud mesenchyme and small cartilaginous condensations, and syndactyly is associated with a complete absence of interdigital cell death. Although Myc family proteins have pro-apoptotic activity, N-Myc is not expressed in interdigital cells undergoing programmed cell death. We provide evidence indicating that the lack of interdigital cell death and associated syndactyly is related to an absence of interdigital cells marked by expression of Fgfr2 and Msx2. Thus, instead of directly regulating interdigital cell death, we propose that N-Myc is required for the proper generation of undifferentiated mesenchymal cells that become localized to interdigital regions and trigger digit separation when eliminated by programmed cell death. Our results provide new insight into mechanisms that control limb development and suggest that defects in the formation of N-Myc-dependent interdigital tissue may be a root cause of common syndromic forms of syndactyly.


M-CSF induces the stable interaction of cFms with alphaVbeta3 integrin in osteoclasts.
The macrophage colony stimulating factor receptor (cFms) and alpha(V)beta(3) integrin are both abundantly expressed and play critical roles in the differentiation, survival and migration of osteoclasts. We have previously demonstrated that cross-talk between cFms- and alpha(V)beta(3)-mediated signaling pathways regulated the cytoskeletal organization required for osteoclast migration. To investigate the nature of interaction between the two receptors, we sequentially used anion-exchange chromatography and immunoprecipitation to purify alpha(V)beta(3)-associated protein complexes. We have demonstrated that cFms stably associated with alpha(V)beta(3) in osteoclasts during adhesion, and that the association was induced by macrophage colony stimulating factor (M-CSF) stimulation. However, the kinetics of association of alpha(V)beta(3) and cFms did not correlate with the kinetics of tyrosine phosphorylation of cFms. Instead, maximally observed alpha(V)beta(3)/cFms association was after the peak of cFms tyrosine phosphorylation and correlated inversely with the total amount of cFms remaining. Furthermore, the complex containing cFms and alpha(V)beta(3) also contained a number of other signaling molecules including Pyk2, p130(Cas) and c-Cbl, known downstream regulators of the integrin-mediated signaling pathways in osteoclasts. In the presence of M-CSF, co-localization of alpha(V)beta(3) integrin and cFms was identified in the podosomal actin ring of the osteoclast during adhesion on glass. Interestingly, co-localization of both receptors was not found in the sealing zone, but in punctate structures associated with adhesion- or transcytosis-like structures in osteoclasts on bone. Taken together, we suggest that the association of alpha(V)beta(3) and cFms could be the result of signaling following tyrosine phosphorylation of cFms. The recruitment of cFms to alpha(V)beta(3) integrin may be an integral part of a larger signaling complex via which both of adhesion- and growth factor receptors coordinately regulate osteoclast adhesion, motility and membrane trafficking.


Chemical modulation of metallothionein I and III mRNA in mouse brain.
Metallothioneins (MTs) are sulfhydryl-rich proteins. MT-I and MT-II are found in all tissues of the body, while MT-III exists only in brain. Regulation of MT-I and MT-III mRNA was studied in brain and liver of control C57BL/6J mice and mice given chemicals known to increase MT-I, namely, lipopolysaccharide (LPS), zinc chloride (Zn), cadmium chloride (Cd), dexamethasone (Dex), ethanol, and kainic acid (KA). Northern blot analysis revealed that MT-I mRNA levels in liver were induced dramatically (12-27-fold over basal levels) by all of the chemicals, while in brain only LPS produced an increase in MT-I mRNA (2-fold). Interestingly, the MT-I inducers, Cd, Dex, ethanol, and KA, down-regulated brain MT-III mRNA levels by approx. 30%. Because brain is such a heterogenous tissue, in situ hybridization was used to localize MT-I and MT-III mRNA in control and treated mice. MT-I mRNA signal, which was most abundant in the glial cells of the Purkinje cell layer of the cerebellum in control mice, appeared to be enhanced in mice given the MT-I inducers (LPS, Zn, Cd, Dex, ethanol, and KA). MT-I mRNA hybridization signal was also enhanced in the olfactory bulbs from LPS- and Cd-treated mice, while this signal was present but weak in control brains. MT-III mRNA hybridization signals were localized in hippocampus and co-localized with MT-I message in the glial cells of the Purkinje cell layer of the cerebellum. In addition, diffuse MT-III mRNA signals were visible in areas of the cerebral cortex, and in the molecular layer of the cerebellum. Signals for MT-III in hippocampus appeared to be reduced by KA, Dex and LPS treatment, while in the cortical region, MT-III mRNA signals appeared to be enhanced by KA, Cd, and ethanol treatment. In conclusion, both MT-I and MT-III expression in brain appears to be modulated by exogenous treatment, however, the changes are small in relation to those observed in liver. Chemical-induced alterations of MT mRNA are non-uniform throughout the brain, and thus best studied in a region-specific manner.


Induced nitric oxide synthesis in the gut of Manduca sexta protects against oral infection by the bacterial pathogen Photorhabdus luminescens.
Injecting the insect pathogenic bacterium Photorhabdus luminescens into the blood system of the model lepidopteran insect Manduca sexta induces nitric oxide synthase (NOS) expression in the fat body and blood cells (haemocytes), whereas following oral ingestion of bacteria NOS expression is limited to the gut. We used RNA interference to knock-down expression of NOS throughout the insect. Preventing NOS induction in this way adversely affected the survival of orally infected insects and caused a significant increase in the number of bacteria crossing into the haemolymph. By contrast, knock-down of NOS had no effect on the mortality rate of insects infected with P. luminescens by injection. Pharmacological inhibition of NOS decreased both nitric oxide (NO) levels in the gut wall and survival of orally infected insects, whereas elevation of gut wall NO using an NO donor increased survival of NOS silenced caterpillars. Together, our results imply that induced synthesis of NO is important in mediating insect immune defence against the pathogen by inhibiting transfer of bacteria across the gut wall.


[Comparison of content of total alkaloids and aconitine in Aconitum szechenyianum from different producing areas on basis of macroporous adsorption resin purification technology].
To optimize the conditions of purifying the total alkaloids in Aconitum szechenyianum with macroporous adsorption resin, and compare the content of total alkaloids and aconitine in A. szechenyianum from different producing areas, in order to provide basis for further studies. The orthogonal experiment method was adopted for optimizing the conditions of purifying the total alkaloids in A. szechenyianum with macroporous adsorption resin. The content of total alkaloids and aconitine were determined by using the titration method. The total alkaloids in A. szechenyianum from different producing areas were purified under optimum processing conditions. Aconitine was determined by HPLC. Different processing conditions showed different influences on the purification of total alkaloids. The optimum conditions were resin type HPD-722, ethanol concentration of 80% , and ethanol elution volume of 80 mL x min(-1). The contents of aconitine in A. szechenyianum from different producing areas--Qinghai, Maxianshan, Ningxia and Yongdeng were 0.493 5, 0.883 5, 1.527 8, 1.664 4 mg x g(-1), respectively. The optimum processing conditions used in this essay could be used for purifying the total alkaloids and aconitine. A. szechenyianum from Yongdeng and Gansu contains the highest content of aconitine.