A 7-year old female with arthrogryposis multiplex congenita, Duane retraction syndrome, and Marcus Gunn phenomenon due to a ZC4H2 gene mutation: a clinical presentation of the Wieacker-Wolff syndrome.
Background: Duane retraction syndrome and arthrogryposis multiplex congenita have an incidence of approximately 1:1500-1:3000 live births. However, the association of these two entities with a Marcus-Gunn might be a rare and, until now, under-recognized clinical presentation of the Wieacker-Wolff Syndrome.Patient and methods: We report a 7-year-old female with dysmorphic features, global developmental delay, arthrogryposis multiplex congenita (AMC), Duane retraction syndrome (DRS), and unilateral Marcus Gunn jaw winking.Results: Whole Exome Sequencing showed a de novo premature stop codon in ZC4H2. Extensive genetic and metabolic work was negative otherwise and Brain MRI showed delayed non-specific myelination abnormalities. She continues to have significant delays but does not have regression, seizures or other neurological complications. She has required a multidisciplinary approach for the management of her multiple contractures.Conclusion: This case confirms ZC4H2 as a cause of syndromic DRS and extends the ZC4H2 phenotype to include Marcus Gunn jaw winking.


Identification and biochemical characterization of a second zebrafish autotaxin gene.
Autotaxin (ATX) is a secreted enzyme that produces a bioactive lysophospholipid, lysophosphatidic acid (LPA). ATX plays a role in vascular and neural development in embryos but its mechanisms remain unclear. At the beginning of this study, only one zebrafish atx gene (atxa) was known and had been investigated. In this study, we generated ATX knockout (KO) fish by TALEN targeting atxa. Unexpectedly, atxa KO fish showed neither vascular defects nor reduction of ATX activity, implying the existence of one or more other ATXs in the genome. By a BLAST search using ATXa protein fragments as a query, we found a genomic sequence that closely resembled atxa exons 13, 14 and 15. Consequently, we cloned a cDNA encoding a second zebrafish autotaxin (ATXb), and found that it was transcribed in various tissues. The atxb gene encoded a protein of 832 amino acids (compared to 850 amino acids in ATXa) with 60% amino acid identity to ATXa and clustered with ATXs from other species. A recombinant ATXb protein showed lysophospholipase D (lysoPLD) activities with substrate specificities similar to those of ATXa and mammalian ATXs. These results indicate that ATXb is a second zebrafish ATX, which possibly shares redundant roles with ATXa in embryonic development.


Age-related changes in protein-related epitopes of human articular-cartilage proteoglycans.
Monoclonal antibodies were prepared that recognize different age-related epitopes on proteoglycan subunits of high buoyant density isolated from human epiphysial and articular cartilages. Antibody EFG-4 (IgG1) recognizes a proteinase-sensitive segment associated with the core protein. Antibody BCD-4 (IgG1) reacts with keratan sulphate bound to core protein. Both epitopes are minimally expressed in foetal cartilage and increase with age after birth to become maximally expressed in adult cartilage by about 30 years of age. In contrast, monoclonal antibody alpha HFPG-846 (IgM) recognizes a core-protein-related epitope that is maximally expressed in young foetal cartilage, declines up to birth and thereafter and is almost absent after about 30 years of age. Antibody alpha HFPG-846 was used to isolate by immuno-affinity chromatography two subpopulations of proteoglycan subunits from a 16-year-old-human cartilage proteoglycan subunit preparation. Only the antibody-unbound population showed a significant reaction with antibodies EGF-4 and BCD-4. The amino acid and carbohydrate compositions of these proteoglycan fractions were different, and one (antibody-bound) resembled those of foetal and the other (antibody-unbound) resembled those of adult proteoglycans isolated from 24-27-week-old-foetal and 52-56-year-old-adult cartilage respectively. These observations demonstrate that human cartilages contain at least two chemically and immunochemically distinct populations of proteoglycans, the proportions and content of which are age-dependent. It is likely that these populations represent the products of different genes, though their heterogeneity may be compounded by the result of different post-translation modifications.


Chemical elucidation of acute encephalopathy by ingestion of angel-wing mushroom (Pleurocybella porrigens) - involvement of three constituents in onset.
The mushroom, Pleurocybella porrigens, is widely consumed in Japan; however, in autumn 2004, acute encephalopathy due to ingestion of the mushroom in a large group of patients was reported in Japan. We have continued working on the mushroom to clarify the mechanisms underlying the acute encephalopathy that occurred due to its consumption. The data collected to date have shown that three compounds, pleurocybelline (PC), a Pleurocybella porrigens lectin (PPL), and pleurocybellaziridine (PA), in the mushroom are potentially responsible for the onset of the disease; PC that exhibit lethal activity in mice and PPL formed a complex, and the complex of the two components exhibited proteolytic activity and disrupted the blood-brain barrier. Although PA was not isolated directly from the mushroom, the existence of this compound in the mushroom was predicted. The compound was chemically synthesized and its endogeneity in the mushroom was demonstrated. Furthermore, PA exhibited toxicity to oligodendrocytes.


DNAzyme-Based Biosensors: Immobilization Strategies, Applications, and Future Prospective.
Since their discovery almost three decades ago, DNAzymes have been used extensively in biosensing. Depending on the type of DNAzyme being used, these functional oligonucleotides can act as molecular recognition elements within biosensors, offering high specificity to their target analyte, or as reporters capable of transducing a detectable signal. Several parameters need to be considered when designing a DNAzyme-based biosensor. In particular, given that many of these biosensors immobilize DNAzymes onto a sensing surface, selecting an appropriate immobilization strategy is vital. Suboptimal immobilization can result in both DNAzyme detachment and poor accessibility toward the target, leading to low sensing accuracy and sensitivity. Various approaches have been employed for DNAzyme immobilization within biosensors, ranging from amine and thiol-based covalent attachment to non-covalent strategies involving biotin-streptavidin interactions, DNA hybridization, electrostatic interactions, and physical entrapment. While the properties of each strategy inform its applicability within a proposed sensor, the selection of an appropriate strategy is largely dependent on the desired application. This is especially true given the diverse use of DNAzyme-based biosensors for the detection of pathogens, metal ions, and clinical biomarkers. In an effort to make the development of such sensors easier to navigate, this paper provides a comprehensive review of existing immobilization strategies, with a focus on their respective advantages, drawbacks, and optimal conditions for use. Next, common applications of existing DNAzyme-based biosensors are discussed. Last, emerging and future trends in the development of DNAzyme-based biosensors are discussed, and gaps in existing research worthy of exploration are identified.


Interfacial thermal transport of graphene/β-Ga2O3 heterojunctions: a molecular dynamics study with a self-consistent interatomic potential.
Graphene/β-Ga2O3 heterojunctions are widely used in high-power and high-frequency devices, for which thermal management is vital to the device operation and life. Here we apply molecular dynamics simulation to calculate the interfacial thermal resistance (ITR) between graphene and β-Ga2O3. Based on the rigid ion model, a self-consistent interatomic potential with a set of parameters that can well reproduce the basic physical properties of crystal β-Ga2O3 is fitted. Using this potential, the effects of model size, interface type, temperature, vacancy defects and graphene hydrogenation on the ITR of graphene/β-Ga2O3 heterojunctions are evaluated. The results show that there is no obvious dependence of ITR on the size of graphene and β-Ga2O3. It is reported that the ITR values of the (100), (010) and (001) interfaces are 7.28 ± 0.35 × 10-8 K m2 W-1, 6.69 ± 0.44 × 10-8 K m2 W-1 and 5.22 ± 0.35 × 10-8 K m2 W-1 at 300 K, respectively. Both temperature increase and vacancy defect increase can prompt the energy propagation across graphene/β-Ga2O3 interfaces due to the enhancement of phonon coupling. In addition, graphene hydrogenation provides new channels for in-plane and out-of-plane phonon coupling, and thus reduces the ITR between graphene and β-Ga2O3. This study provides basic strategies for the thermal design and management of graphene/β-Ga2O3 based photoelectric devices.


Targeting apoptosis resistance in rhabdomyosarcoma.
Resistance of human cancers to current treatment approaches remains a challenge in oncology. Therefore, there has been much interest in identifying molecular pathways that are responsible for primary or acquired resistance of cancers. Since most anticancer therapies, i.e. chemotherapy or radiotherapy, primarily act by triggering programmed cell death (apoptosis) in cancer cells, defects in apoptosis programs may confer resistance. Evasion of apoptosis in rhabdomyosarcoma may be caused by the dominance of cell survival pathways, for example aberrant activation of the PI3K/Akt/mTOR cascade, or alternatively, by defective expression or function of critical mediators of apoptosis, i.e. components of the TRAIL signaling system. In addition, signaling to apoptosis can be blocked under hypoxia, a characteristic feature of most solid tumors including rhabdomyosarcoma that has been associated with poor treatment response. Thus, molecular targeted therapies that are specifically directed to the defects in apoptosis programs, open novel perspectives to restore apoptosis sensitivity in rhabdomyosarcoma.


Redox regulation and flower development: a novel function for glutaredoxins.
Glutaredoxins (GRXs) are small, ubiquitous oxidoreductases that have been intensively studied in E. COLI, yeast and humans. They are involved in a large variety of cellular processes and exert a crucial function in the response to oxidative stress. GRXs can reduce disulfides by way of conserved cysteines, located in conserved active site motifs. As in E. COLI, yeast, and humans, GRXs with active sites of the CPYC and CGFS type are also found in lower and higher plants, however, little has been known about their function. Surprisingly, 21 GRXs from ARABIDOPSIS THALIANA contain a novel, plant-specific CC type motif. Lately, information on the function of CC type GRXs and redox regulation, in general, is accumulating. This review focuses on recent findings indicating that GRXs, glutathione and redox regulation, in general, seem to be involved in different processes of development, so far, namely in the formation of the flower. Recent advances in EST and genome sequencing projects allowed searching for the presence of the three different types of the GRX subclasses in other evolutionary informative plant species. A comparison of the GRX subclass composition from PHYSCOMITRELLA, PINUS, ORYZA, POPULUS, and ARABIDOPSIS is presented. This analysis revealed that only two CC type GRXs exist in the bryophyte PHYSCOMITRELLA and that the CC type GRXs group expanded during the evolution of land plants. The existence of a large CC type subclass in angiosperms supports the assumption that their capability to modify target protein activity posttranslationally has been integrated into crucial plant specific processes involved in higher plant development.


Cholesterol loss enhances TrkB signaling in hippocampal neurons aging in vitro.
Binding of the neurotrophin brain-derived neurotrophic factor (BDNF) to the TrkB receptor is a major survival mechanism during embryonic development. In the aged brain, however, BDNF levels are low, suggesting that if TrkB is to play a role in survival at this stage additional mechanisms must have developed. We here show that TrkB activity is most robust in the hippocampus of 21-d-old BDNF-knockout mice as well as in old, wild-type, and BDNF heterozygous animals. Moreover, robust TrkB activity is evident in old but not young hippocampal neurons differentiating in vitro in the absence of any exogenous neurotrophin and also in neurons from BDNF -/- embryos. Age-associated increase in TrkB activity correlated with a mild yet progressive loss of cholesterol. This, in turn, correlated with increased expression of the cholesterol catabolic enzyme cholesterol 24-hydroxylase. Direct cause-effect, cholesterol loss-high TrkB activity was demonstrated by pharmacological means and by manipulating the levels of cholesterol 24-hydroxylase. Because reduced levels of cholesterol and increased expression of choleseterol-24-hydroxylase were also observed in the hippocampus of aged mice, changes in cellular cholesterol content may be used to modulate receptor activity strength in vivo, autonomously or as a way to complement the natural decay of neurotrophin production.


Ambient air pollution and the progression of atherosclerosis in adults.
Cross-sectional studies suggest an association between exposure to ambient air pollution and atherosclerosis. We investigated the association between outdoor air quality and progression of subclinical atherosclerosis (common carotid artery intima-media thickness, CIMT). We examined data from five double-blind randomized trials that assessed effects of various treatments on the change in CIMT. The trials were conducted in the Los Angeles area. Spatial models and land-use data were used to estimate the home outdoor mean concentration of particulate matter up to 2.5 micrometer in diameter (PM2.5), and to classify residence by proximity to traffic-related pollution (within 100 m of highways). PM2.5 and traffic proximity were positively associated with CIMT progression. Adjusted coefficients were larger than crude associations, not sensitive to modelling specifications, and statistically significant for highway proximity while of borderline significance for PM2.5 (P = 0.08). Annual CIMT progression among those living within 100 m of a highway was accelerated (5.5 micrometers/yr [95%CI: 0.13-10.79; p = 0.04]) or more than twice the population mean progression. For PM2.5, coefficients were positive as well, reaching statistical significance in the socially disadvantaged; in subjects reporting lipid lowering treatment at baseline; among participants receiving on-trial treatments; and among the pool of four out of the five trials. Consistent with cross-sectional findings and animal studies, this is the first study to report an association between exposure to air pollution and the progression of atherosclerosis--indicated with CIMT change--in humans. Ostensibly, our results suggest that air pollution may contribute to the acceleration of cardiovascular disease development--the main causes of morbidity and mortality in many countries. However, the heterogeneity of the volunteering populations across the five trials, the limited sample size within trials and other relevant subgroups, and the fact that some key findings reached statistical significance in subgroups rather than the sample precludes generalizations to the general population.


Subsurface damage in some single crystalline optical materials.
We present a nondestructive method for estimating the depth of subsurface damage (SSD) in some single crystalline optical materials (silicon, lithium niobate, calcium fluoride, magnesium fluoride, and sapphire); the method is established by correlating surface microroughness measurements, specifically, the peak-to-valley (p-v) microroughness, to the depth of SSD found by a novel destructive method. Previous methods for directly determining the depth of SSD may be insufficient when applied to single crystals that are very soft or very hard. Our novel destructive technique uses magnetorheological finishing to polish spots onto a ground surface. We find that p-v surface microroughness, appropriately scaled, gives an upper bound to SSD. Our data suggest that SSD in the single crystalline optical materials included in our study (deterministically microground, lapped, and sawed) is always less than 1.4 times the p-v surface microroughness found by white-light interferometry. We also discuss another way of estimating SSD based on the abrasive size used.


Pulmonary embolization of papillary fibroelastoma. Arising from the tricuspid valve.
Papillary fibroelastomas are rare, benign tumors of the heart that often are incidental findings at autopsy or cardiac surgery, or during echocardiography. These tumors are predisposed to valvular involvement, and all such lesions detected by echocardiography have occurred in the left side of the heart, except for 3 on the tricuspid valve. Left-sided lesions have been associated with embolic phenomena; however, embolization of right-sided lesions has never been documented. We report a tricuspid valve fibroelastoma detected by 2-dimensional and transesophageal echocardiography, and confirmed by surgical excision and biopsy. To our knowledge, this is the 1st reported case of pulmonary embolization of papillary fibroelastoma arising from the tricuspid valve.


Mitochondrial translocation of vitamin D receptor is mediated by the permeability transition pore in human keratinocyte cell line.
Vitamin D receptor (VDR) is a well known transcriptional regulator, active as heterodimer in association with coactivators and corepressors. In addition it has been described the extranuclear distribution of the receptor and in particular the recently reported mitochondrial localization in platelets and megakaryocytes is intriguing because it appears to be a common feature of steroid receptors. Whereas for other members of the steroid receptor family the mitochondrial function has been explored, up to now nothing is known about a mitochondrial form of VDR in human proliferating cells. In this study we characterized for the first time the mitochondrial localization of VDR in the human keratinocyte cell line HaCaT. In proliferating HaCaT cells VDR was abundantly expressed in mitochondria in association with its binding partner RXRα and the import was ligand-independent. By immunoprecipitation studies we demonstrated the interaction of VDR with proteins of the permeability transition pore (PTP), VDAC and StAR. We then adopted different pharmacological and silencing approaches with the aim of hampering PTP function, either affecting PTP opening or abating the expression of the complex member StAR. By all means the impairment of pore function led to a reduction of mitochondrial levels of VDR. The results reported here demonstrate a ligand-independent mitochondrial import of VDR through the permeability transition pore, and open interesting new perspectives on PTP function as transporter and on VDR role in mitochondria.


Application of Machine Learning and Grocery Transaction Data to Forecast Effectiveness of Beverage Taxation.
Sugar Sweetened Beverages (SSB) are the primary source of artificially added sugar and have a casual association with chronic diseases. Taxation of SSB has been proposed, but limited evidence exists to guide this public health policy. Grocery transaction data, with price, discounting and other information for beverage products, present an opportunity to evaluate the likely effects of taxation policy. Sales are often non-linearly associated with price and are affected by the prices of multiple competing brands. We evaluated the predictive performance of Boosted Decision Tree Regression (B-DTR) and Deep Neural Networks (DNN) that account for the non-linearity and competition across brands, and compared their performance to a benchmark regression, the Least Absolute Shrinkage and Selection Operator (LASSO). B-DTR and DNN showed a lower Mean Squared Error (MSE) of prediction in the sales of most major SSB brands in comparison to LASSO, indicating a superior accuracy in predicting the effectiveness of SSB taxation. We demonstrated the application of machine learning methods and large transactional data from grocery stores to forecast the effectiveness food taxation.


Efficacy of methylphenidate treatment in childhood myotonic dystrophy type 1 and comorbid attention deficit hyperactivity disorder: A case report using eye tracking assessment.
Despite the increased prevalence of comorbid attention deficit hyperactivity disorder (ADHD) in children with myotonic dystrophy type 1, the effects of methylphenidate treatment on associated cognitive deficits in this population is not yet investigated. We describe a case study of an eleven-year-old male patient with myotonic dystrophy type 1 and comorbid ADHD that was treated with methylphenidate in a twice daily regime (0.60 mg/kg/day). Positive effects on learning and cognition were reported by the parents and teachers. No negative side effects were reported. Sequential neuropsychological assessments before and 45 minutes after methylphenidate intake were conducted to quantify the cognitive effects of methylphenidate treatment. Significant improvements in regulation of attention were behaviorally observed and were quantified using eye tracking technology. We conclude that methylphenidate may be an effective treatment for ADHD-related cognitive deficits and learning difficulties in children with myotonic dystrophy type 1 which merits further research.


Adherence to clinical practice guidelines (CPGs) for the treatment of cancers in Australia and the factors associated with adherence: a systematic review protocol.
Clinical practice guidelines (CPGs) synthesise the latest evidence to support clinical and patient decision-making. CPG adherent care is associated with improved patient survival outcomes; however, adherence rates are low across some cancer streams in Australia. Greater understanding of specific barriers to cancer treatment CPG adherence is warranted to inform future implementation strategies.This paper presents the protocol for a systematic review that aims to determine cancer treatment CPG adherence rates in Australia across a variety of common cancers, and to identify any factors associated with adherence to those CPGs, as well as any associations between CPG adherence and patient outcomes. Five databases will be searched, Ovid Medline, PsychInfo, Embase, Scopus and Web of Science, for eligible studies evaluating adherence rates to cancer treatment CPGs in Australia. A team of reviewers will screen the abstracts in pairs according to predetermined inclusion criteria and then review the full text of eligible studies. All included studies will be assessed for quality and risk of bias. Data will be extracted using a predefined data extraction template. The frequency or rate of adherence to CPGs, factors associated with adherence to those CPGs and any reported patient outcome rates (eg, relative risk ratios or 5-year survival rates) associated with adherence to CPGs will be described. If applicable, a pooled estimate of the rate of adherence will be calculated by conducting a random-effects meta-analysis. The systematic review will adhere to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. Ethics approval will not be required, as this review will present anonymised data from other published studies. Results from this study will form part of a doctoral dissertation (MB), will be published in a journal, presented at conferences, and other academic presentations. CRD42020222962.


Noninvasive measurements of nonlinear arterial elasticity.
To describe fully the compliance characteristics of an artery with nonlinear elastic properties, measurements must be obtained over a wide range of pressures. Furthermore, repeated measurements, as required in temporal studies of arterial implants, require that the measuring technique be noninvasive. The application of a pulsed ultrasound echo-tracking device is described, which fulfills both criteria. Nonlinear compliance-pressure (CP) curves were obtained from the femoral arteries of dogs, with the use of halothane anesthesia to vary systemic pressure, and were used to compare the gross elastic properties of different vessels. Studies using controlled hemorrhage or the vasoactive drugs, nitroprusside and levarterenol (norepinephrine), were used to verify that the CP curves obtained during halothane anesthesia did not reflect varying degrees of smooth muscle activation. However, surgical exposure did temporarily reduce arterial compliance at pressures between 60 and 140 mmHg. The effect of vasoactive intervention and of postsurgical changes in arterial or graft compliance can thus be quantitated by use of CP curves or by comparing incremental elastic moduli, which can also be estimated from the noninvasively derived measurements.


Phosphorylation of angiomotin by Lats1/2 kinases inhibits F-actin binding, cell migration, and angiogenesis.
The Hippo tumor suppressor pathway plays important roles in organ size control through Lats1/2 mediated phosphorylation of the YAP/TAZ transcription co-activators. However, YAP/TAZ independent functions of the Hippo pathway are largely unknown. Here we report a novel role of the Hippo pathway in angiogenesis. Angiomotin p130 isoform (AMOTp130) is phosphorylated on a conserved HXRXXS motif by Lats1/2 downstream of GPCR signaling. Phosphorylation disrupts AMOT interaction with F-actin and correlates with reduced F-actin stress fibers and focal adhesions. Furthermore, phosphorylation of AMOT by Lats1/2 inhibits endothelial cell migration in vitro and angiogenesis in zebrafish embryos in vivo. Thus AMOT is a direct substrate of Lats1/2 mediating functions of the Hippo pathway in endothelial cell migration and angiogenesis.


[Assessment of nutritional risks among elderly hospitalized patients - which body mass index and degree of weight loss should be considered?].
The "Subjective Global Assessment", the "Nutrition Risk Score" and a Danish counterpart are simple screening methods to detect patients at risk of nutrition-related complications. The cut-off points used in the screening are a body mass index (BMI) less than 20 kg/m2 (or 18.5 kg/m2 in the Danish version) and weight loss of more than 5% during the last month--regardless of age. This review of the literature indicates that the optimal range of BMI for elderly people is increasing from 18.7-25 kg/m2 to 24-29 kg/m2. It also suggests that a clinically significant weight loss for the elderly is around 5% annually, less than in younger age groups. Finally, there seems to be a high percentage of the old, especially those initially malnourished, who will not benefit from nutritional support. In conclusion, other cut-off points should be used for elderly (65+ years) patients, i.e. BMI less than 24 kg/m2 or any degree of weight loss.


[The activity of the sodium-potassium pump of the mesenteric arteries and veins in normal rats and in DOCA-salt hypertension].
The activity of sodium-potassium pump was studied in mesenteric arteries and veins of rats, as well as its inhibition effect upon catecholamine-induced contractile responses and the time of relaxation of these vessels in normal rats and in those with DOCA-salt hypertension. The activity was measured by 86Rb [correction of 86Rh] up-take. A difference in the ouabain-sensitive take-up was found between arteries and veins. It was higher in normotensive rats' veins than in arteries; in hypertension, the activity of the pump tended to decrease in arteries and sharply increased in veins. Inhibition of the pump with ouabain reduced arteries' contractile responses to noradrenaline and phenylephrine in normotensive animals, whereas in hypertension the effect involved all the types of stimulation leaving intact the contractions of venous vessels. Ouabain increased the period of relaxation of arteries and veins in some types of stimulation in normal as well as in hypertensive rats.


Ethanol exposure represses osteogenesis in the developing chick embryo.
It is known that excess alcohol consumption during pregnancy can increase the risk of fetal alcohol spectrum disorder (FASD). However, the effect of ethanol exposure on bone morphogenesis in fetus is largely unknown. In this study, we demonstrated that ethanol treatment of gastrulating chick embryos could inhibit long bone (humerus, radius and ulna) development. Histological examination revealed that ethanol exposure reduced the width of the proliferation and hypertrophic zones. In addition, cell proliferation and alkaline phosphatase activities were repressed. We also investigated the effect on chondrogenesis and chondrogenesis was inhibited. Ethanol exposure also induced excess reactive oxygen species (ROS) production and altered the expression of osteogenesis-related genes. The inhibiting effect on flat bone (sclerotic ossicle) and the generation of cranial neural crest cells (progenitors of craniofacial bones) was also presented. In conclusion, ethanol exposure during the embryonic period retards bone development through excess ROS production and altered bone-associated gene expression.


Collecting, shipping, storing, and imaging snow crystals and ice grains with low-temperature scanning electron microscopy.
Methods to collect, transport, and store samples of snow and ice have been developed that enable detailed observations of these samples with a technique known as low-temperature scanning electron microscopy (LTSEM). This technique increases the resolution and ease with which samples of snow and ice can be observed, studied, and photographed. Samples are easily collected in the field and have been shipped to the electron microscopy laboratory by common air carrier from distances as far as 5,000 miles. Delicate specimens of snow crystals and ice grains survive the shipment procedures and have been stored for as long as 3 years without undergoing any structural changes. The samples are not subjected to the melting or sublimation artifacts. LTSEM allows individual crystals to be observed for several hours with no detectable changes. Furthermore, the instrument permits recording of photographs containing the parallax information necessary for three-dimensional imaging of the true shapes of snowflakes, snow crystals, snow clusters, ice grains, and interspersed air spaces. This study presents detailed descriptions of the procedures that have been used successfully in the field and the laboratory to collect, ship, store, and image snow crystals and ice grains.


Emerging risk factors for cerebrovascular disease.
Nontraditional risk factors for cerebrovascular disease are rapidly emerging. The categories are expanding, and include those related to infection, inflammation, sleep disorders, hemostasis, nutrition, endocrine, and one's individual genotype. Many of the promising factors lack large randomized prospective population studies confirming direct cause and effect. However there have been strong evidence supporting increased stroke risk factor for infection, obstructive sleep disorders, the metabolic syndrome and impaired glucose tolerance in particular. Unique drug targets have already been identified in some of these emerging risk factors. The complexity of the pathophysiology of this disease remains a challenge. For example despite repeated evidence of estrogen-related neuroprotection, large population-based studies in postmenopausal women receiving estrogen replacement did not demonstrate the expected neuroprotection. This suggests that aggressive research both at the basic science and transitional level needs to evolve, to ensure targeted successful stroke therapy. The advent of nanotechnology including the development of targeted therapeutic nanospheres, and of revolutionary molecular technology resulting in the synthesis of specific peptide mimetics, bodes well for the future development of cerebrovascular drug treatment.


Widespread microplastic ingestion by fish assemblages in tropical estuaries subjected to anthropogenic pressures.
Our aim was to quantify microplastic ingestion by fish assemblages in two tropical Brazilian estuaries and to evaluate whether biological and ecological factors influence the ingestion of microplastics by fish species. Of 2233 fish from both estuaries (from 69 species) examined in this study, 9% of the individuals (24 species) had microplastics in their gut contents. Microplastic ingestion occurred irrespective of fish size and functional group. The diet of fish species was analyzed based on prey items identified in the fish's full stomach contents and five feeding guilds were defined. Microplastics were common throughout all feeding guilds. Low (average ingestion values 1.06±0.30 items/total fish) but widespread occurrence among estuaries also indicates proliferation of microplastic pollution. Our findings highlight the need to focus on assemblage level studies to understand the real magnitude of the problem and emphasize the urgency of mitigation measures directed at microplastic pollution in estuarine ecosystems.


Laser treatment of peri-implantitis: a literature review.
Peri-implantitis is a state defined as an inflammatory reaction around osseointegrated implants, leading to progressive loss of supporting bone. Various treatment methods are suggested in the treatment of peri-implantitis and clinicians have to choose a method over a large number of treatment protocols. Lasers have shown promising therapeutic effect in treatment of peri-implantitis. However, some controversies have been found in clinical outcomes after using lasers. Therefore, we aimed to review the current literature over the past ten years for the use of lasers in treatment of peri-implantitis, via the Pubmed electronic database of the US National Library of Medicine. Fifteen human studies were reviewed. Er:YAG (Erbium-Doped Yttrium Aluminum Garnet), CO2(Carbon Dioxide Laser) and Diode lasers were used. Despite inconsistencies and disharmonies among studies in terms of study design, positive treatment outcomes were obvious among the majority of them. However, short period of follow-ups and poor control of plaque index, as a critical confounding factor, were the major problems which these studies suffered from. It seems that one session laser therapy is not adequate for achieving optimal clinical outcome. Further studies with longer periods of follow-ups, intense control of plaque index, and various sessions of laser treatments are needed to clearly illustrate the clinical privilege of laser therapy.


Mgm101: A double-duty Rad52-like protein.
Mgm101 has well-characterized activity for the repair and replication of the mitochondrial genome. Recent work has demonstrated a further role for Mgm101 in nuclear DNA metabolism, contributing to an S-phase specific DNA interstrand cross-link repair pathway that acts redundantly with a pathway controlled by Pso2 exonuclease. Due to involvement of FANCM, FANCJ and FANCP homologues (Mph1, Chl1 and Slx4), this pathway has been described as a Fanconi anemia-like pathway. In this pathway, Mgm101 physically interacts with the DNA helicase Mph1 and the MutSα (Msh2/Msh6) heterodimer, but its precise role is yet to be elucidated. Data presented here suggests that Mgm101 functionally overlaps with Rad52, supporting previous suggestions that, based on protein structure and biochemical properties, Mgm101 and Rad52 belong to a family of proteins with similar function. In addition, our data shows that this overlap extends to the function of both proteins at telomeres, where Mgm101 is required for telomere elongation during chromosome replication in rad52 defective cells. We hypothesize that Mgm101 could, in Rad52-like manner, preferentially bind single-stranded DNAs (such as at stalled replication forks, broken chromosomes and natural chromosome ends), stabilize them and mediate single-strand annealing-like homologous recombination event to prevent them from converting into toxic structures.


Design and synthesis of highly potent and selective melanotropin analogues of SHU9119 modified at position 6.
The melanocortin receptors are involved in several important physiological functions. The potent and enzymatically stable analogues MT-II (Ac-Nle-c[Asp-His-DPhe-Arg-Trp-Lys]-NH(2)) and SHU9119 (Ac-Nle-c[Asp-His-DNal(2')-Arg-Trp-Lys]-NH(2)) are important ligands of these receptors but are relatively nonselective. To differentiate between the physiological functions of these receptors, agonists, and antagonists with improved receptor selectivities are needed. We report here analogues of the well-characterized antagonist SHU9119 in which we replaced His(6) with conformationally constrained amino acids. By this structure-activity study we discovered two important compounds, PG-901 (Ac-Nle(4)-c[Asp(5)-Pro(6)-DNal(2')(7)-Arg(8)-Trp(9)-Lys(10)]-NH(2)) and PG-911 (Ac-Nle(4)-c[Asp(5)-Hyp(6)-DNal(2')(7)-Arg(8)-Trp(9)-Lys(10)]-NH(2)), characterized to be full agonists at the hMC5R (EC(50) = 0.072 nM and 0.031 nM, respectively), but full antagonists at the hMC3R and the hMC4R. We also demonstrated that the relative stereochemistry of the amino acid at the 6-position is critical for activity, and could play an important role in potency as well as in selectivity for the melanocortin receptors.


Advances in diagnosis, classification, pathophysiology, and management of trigeminal neuralgia.
Trigeminal neuralgia is a very painful neurological condition with severe, stimulus-evoked, short-lasting stabbing pain attacks in the face. The past decade has offered new insights into trigeminal neuralgia symptomatology, pathophysiology, and treatment, leading to a change in the classification of the condition. An accurate diagnosis is crucial because neuroimaging interpretation and clinical management differ among the various forms of facial pain. MRI using specific sequences should be a part of the diagnostic workup to detect a possible neurovascular contact and exclude secondary causes. Demonstration of a neurovascular contact should not be used to confirm a diagnosis but rather to facilitate surgical decision making. Carbamazepine and oxcarbazepine are drugs of first choice for long-term treatment, whereas microvascular decompression is the first-line surgery in medically refractory patients. Advances in neuroimaging techniques and animal models will provide further insight into the causes of trigeminal neuralgia and its pathophysiology. Development of more efficacious treatment options is highly warranted.


The day the earth moved.
Diane Lowder wasn't sure how ready her hospital was for a catastrophe. But a unique disaster plan helped keep Northridge Hospital open after the '94 Los Angeles earthquake. Here's what Lowder saw at the epicenter.


Protein kinase C-alpha participates in FcgammaR-mediated phagocytosis in macrophages.
Phagocytosis of IgG-opsonized particles by macrophages requires the activation of protein kinase C (PKC), a family of protein serine/threonine kinases. In the present study, we have investigated the role of the PKC-alpha isoenzyme in FcgammaR-mediated phagocytosis using clones of the mouse macrophage cell line RAW 264. 7 overexpressing a dominant-negative (DN) mutant of PKC-alpha. Overexpression of DN PKC-alpha had no effect on the attachment of IgG-opsonized sheep red blood cells, but inhibited their internalization. Further analysis of the signaling events induced by IgG-opsonized sheep red blood cells revealed that whereas tyrosine phosphorylation of Syk was normal, phosphorylation of ERK 1/2 (p42/44) was impaired in DN PKC-alpha-overexpressing macrophages. These observations suggest a role for PKC-alpha in the regulation of FcgammaR-induced phagocytosis and signal transduction.


Soft and hard tissue management in implant therapy-part I: surgical concepts.
Implant therapy has become a reliable and predictable treatment alternative for the replacement of missing teeth with conventional removable and fixed partial dentures. Recently though, in the pursuit for improved esthetics, the literature has dedicated a considerable amount of its research on the successful maintenance and regeneration of the surrounding gingiva and bone, which are lost following extraction of a tooth. Thoroughly analyzing the anatomic situation and well-planned treatment has become a requirement, because incorrectly planned and positioned implants may jeopardize long-term esthetic and functional prognosis. In addition, many types of biocompatible materials, autogenous hard and soft tissue grafts, and different surgical techniques have been developed, and their viability has been investigated. As a result, implant specialists have gained a greater understanding of the dynamics and anatomical and biological concepts of the periodontium and peri-implant tissues both at the surgical and prosthetic phases of treatment, which contributes to better soft and hard tissue management (SHTM). This may further contribute to achieving a superior final result which is obtained by having a harmonious soft tissue profile, a correctly placed and contoured final restoration, and the reestablishment of masticatory function and phonetics.


Prevalence and severity of atopic dermatitis in Jeju Island: a cross-sectional study of 4,028 Korean elementary schoolchildren by physical examination utilizing the three-item severity score.
The objective of this study was to evaluate the precise prevalence of atopic dermatitis (AD) in schoolchildren in Jeju Island in South Korea examined in 2009. Nine elementary schools were randomly selected from Jeju Island and a total of 4,028 schoolchildren were examined by a dermatologist. AD was diagnosed based on the Korean Atopic Dermatitis Research Group criteria for the disease. The severity of AD was measured with the three-item severity score (TIS). The point prevalence of AD was 9.5% overall. The prevalence among higher graders (age 9-12 years) was significantly lower than that in lower graders (age 6-9 years) (7.5% vs. 11.9%, < 0.00001). AD prevalence in girls (11.1%) was higher than that in boys (8.1%) (<0.005). In each grade, more than 50% of those affected had the mild form (TIS score 1 or 2). There were no apparent differences in severity of AD between grades or genders. This is the first Asian study of prevalence in schoolchildren using TIS score for evaluating AD severity.


Functional role of voltage gated Ca(2+) channels in heart automaticity.
Pacemaker activity of automatic cardiac myocytes controls the heartbeat in everyday life. Cardiac automaticity is under the control of several neurotransmitters and hormones and is constantly regulated by the autonomic nervous system to match the physiological needs of the organism. Several classes of ion channels and proteins involved in intracellular Ca(2+) dynamics contribute to pacemaker activity. The functional role of voltage-gated calcium channels (VGCCs) in heart automaticity and impulse conduction has been matter of debate for 30 years. However, growing evidence shows that VGCCs are important regulators of the pacemaker mechanisms and play also a major role in atrio-ventricular impulse conduction. Incidentally, studies performed in genetically modified mice lacking L-type Cav1.3 (Cav1.3(-/-)) or T-type Cav3.1 (Cav3.1(-/-)) channels show that genetic inactivation of these channels strongly impacts pacemaking. In cardiac pacemaker cells, VGCCs activate at negative voltages at the beginning of the diastolic depolarization and importantly contribute to this phase by supplying inward current. Loss-of-function of these channels also impairs atrio-ventricular conduction. Furthermore, inactivation of Cav1.3 channels promotes also atrial fibrillation and flutter in knockout mice suggesting that these channels can play a role in stabilizing atrial rhythm. Genomic analysis demonstrated that Cav1.3 and Cav3.1 channels are widely expressed in pacemaker tissue of mice, rabbits and humans. Importantly, human diseases of pacemaker activity such as congenital bradycardia and heart block have been attributed to loss-of-function of Cav1.3 and Cav3.1 channels. In this article, we will review the current knowledge on the role of VGCCs in the generation and regulation of heart rate and rhythm. We will discuss also how loss of Ca(2+) entry through VGCCs could influence intracellular Ca(2+) handling and promote atrial arrhythmias.


Androgen priming and chemotherapy in advanced prostate cancer: evaluation of determinants of clinical outcome.
We conducted a randomized clinical trial in men with stage D2 prostate cancer to test whether androgen priming potentiates the efficacy of cytotoxic chemotherapy. Eighty-five men with progressive prostate cancer refractory to orchiectomy were treated continuously with aminoglutethimide and hydrocortisone to lower adrenal androgen secretion and were administered cyclic intravenous (IV) chemotherapy. The patients were randomized to receive either androgen priming or no additional treatment for three days before and on the day of chemotherapy. Median duration of follow-up was 43 months. Response rate (remission plus disease stabilization) was not significantly different between the stimulation and control arm when the analysis was restricted to evaluable patients (79% v 73%, respectively) or when it was extended to all patients (46% v 61%). Median duration of response was similar for the stimulation and control arm (9 and 10 months, respectively). Median survival was 10 months in the stimulation and 15 months in the control group (P = .0047). The androgen sensitivity of the tumors was supported by the greater toxicity in the stimulation arm associated with androgen administration. Factors found to be independently associated with improved clinical outcome included a high Karnofsky score and hematocrit, long duration of response to the initial castration, and normalization of an elevated serum acid phosphatase on treatment. We conclude that in this group of patients with advanced disease, androgen priming does not potentiate the efficacy of chemotherapy and is actually associated with a worse outcome. Furthermore, our data emphasize the heterogeneity of biologic behavior of prostate cancer.


Two-photon Calcium Imaging in Neuronal Dendrites in Brain Slices.
Calcium (Ca2+) imaging is a powerful tool to investigate the spatiotemporal dynamics of intracellular Ca2+ signals in neuronal dendrites. Ca2+ fluctuations can occur through a variety of membrane and intracellular mechanisms and play a crucial role in the induction of synaptic plasticity and regulation of dendritic excitability. Hence, the ability to record different types of Ca2+ signals in dendritic branches is valuable for groups studying how dendrites integrate information. The advent of two-photon microscopy has made such studies significantly easier by solving the problems inherent to imaging in live tissue, such as light scattering and photodamage. Moreover, through combination of conventional electrophysiological techniques with two-photon Ca2+ imaging, it is possible to investigate local Ca2+ fluctuations in neuronal dendrites in parallel with recordings of synaptic activity in soma. Here, we describe how to use this method to study the dynamics of local Ca2+ transients (CaTs) in dendrites of GABAergic inhibitory interneurons. The method can be also applied to studying dendritic Ca2+ signaling in different neuronal types in acute brain slices.


Metabolic regulation of the pancreatic beta-cell ATP-sensitive K+ channel: a pas de deux.
Closure of ATP-sensitive K+ channels (KATP channels) is a key step in glucose-stimulated insulin secretion. The precise mechanism(s) by which glucose metabolism regulates KATP channel activity, however, remains controversial. It is widely believed that the principal determinants are the intracellular concentrations of the metabolic ligands, ATP and ADP, which have opposing actions on KATP channels, with ATP closing and MgADP opening the channel. However, the sensitivity of the channel to these nucleotides in the intact cell, and their relative contribution to the regulation of channel activity, remains unclear. The precise role of phosphoinositides and long-chain acyl-CoA esters, which are capable of modulating the channel ATP sensitivity, is also uncertain. Furthermore, it is still a matter of debate whether it is changes in the concentration of ATP, of MgADP, or of other agents, which couples glucose metabolism to KATP channel activity. In this article, we review current knowledge of the metabolic regulation of the KATP channel and provide evidence that MgADP (or MgATP hydrolysis), acting at the regulatory subunit of the channel, shifts the ATP concentration-response curve into a range in which the channel pore can respond to dynamic changes in cytosolic ATP. This metabolic pas de deux orchestrates the pivotal role of ATP in metabolic regulation of the KATP channel.


Medial collateral knee ligament healing. Combined medial collateral and anterior cruciate ligament injuries studied in rabbits.
We examined the histological appearance and biochemical properties of the healing medial collateral ligament (MCL) of a rabbit knee after combined MCL and anterior cruciate ligament (ACL) injury treated with ACL reconstruction and with or without MCL repair. By so doing, we hoped to understand better our previous bomechanical observations (Ohno et al. 1995) and possibly learn where to focus future investigation into improving the quality of the healing MCL. Ligaments were examined at 6 and 12 weeks of healing. We found healing of all ligaments with hypercellularity and fibroblast elongation along the axis of loading, as expected. Unexpected, however, was the finding of multiple osteophytes in both the repaired and nonrepaired specimens at the medial borders of the joint and at the MCL insertions. These were felt to affect possibly the biomechanics of the MCL by causing stress risers at the point where they undermine the ligament. Biochemically, we demonstrated a correlation between collagen content and hydroxypyridinium crosslinks and modulus of elasticity. While this implies that the modulus is dependent on collagen content and hydroxypyridinium crosslink density, modulus is also probably dependent on other factors such as collagen organization, type and internal structure. Overall, the detailed characterization and correlation between the histological, biochemical, and biomechanical properties of the healing MCL in the severe knee injury model provide insight into the functional behavior of the healing MCL.


Cost analysis of store-and-forward telepsychiatry as a consultation model for primary care.
Store-and-forward telepsychiatry, or asynchronous telepsychiatry (ATP), which allows clinical data, including video to be collected to be reviewed at a later time by a specialist, has been described as a feasible alternative to real-time telepsychiatry, or synchronous telepsychiatry (STP), as a consultation model for primary care. In theory, ATP should be economically more cost-effective than STP due to the increased flexibility of patient data collection and the substitution of the time of specialists with that of lower-cost providers. The aim of this study was to conduct a retrospective cost-analysis comparing ATP with STP and traditional in-person psychiatric consultations in the primary care setting. One hundred and twenty five ATP consultations were performed and fixed and marginal costs were calculated for each model using inputs such as equipment costs, time spent by providers and support staff, and hourly salaries. The fixed costs were $7,000 and $20,000 for ATP and STP and marginal costs were $68.18, $107.50, and $96.36, respectively, for the three groups. STP was the most expensive of the three types of consultations. ATP became the most cost-effective of the three models beyond 249 consultations. The marginal cost savings of ATP were due to substitution of low-cost providers for specialists. ATP represents a potential disruptive healthcare process that could allow more affordable care to be delivered to a larger population of patients. A full accounting of ATP's efficiency will require further studies, including prospective cost-benefit analyses from the perspectives of the patient, provider, and society.


The Content and Accessibility of Orthopaedic Residency Program Websites.
Applicants to orthopaedic surgery residency programs face a competitive match. Internet resources such as program websites allow prospective applicants to gauge interest in particular programs. This study evaluated the content and accessibility of orthopaedic surgery residency program websites. Existing orthopaedic surgery residency programs for the 2020 application cycle were identified on the Electronic Residency Application Service (ERAS) website. Individual program websites were accessed through links directly from the ERAS website, and a Google search for each program was performed to corroborate accessibility. Programs websites were then reviewed and evaluated on the presence of 20 criteria selected for their potential to influence resident recruitment (10) and education (10), respectively. The results were compared with the lone 2001 study and with orthopaedic fellowship website analyses. One hundred eighty-nine orthopaedic surgery residency programs were accredited at the time of the study. Only 6 programs (3.2%) did not have a website identifiable through ERAS or Google searches, leaving a final sample size of 183 websites. Approximately 73.3% of all recruitment content and 44.9% of education content were present on the websites available. There was a significant increase in all available recruitment and education content (p < 0.05) when compared with the lone 2001 study. Orthopaedic residency program websites provide comparable recruitment content at a higher rate (71.1%) than orthopaedic fellowship websites (59.6%) but fall slightly below average in presentation of education content (44.9% vs 45.9%). This is the first study in nearly 20 years to assess the content and accessibility of orthopaedic residency program websites. There is noticeable variability in the presentation of website content, but approximately 73.3% of recruitment content and 44.9% of the educational content were easily accessible through internet search. Orthopaedic surgery residency programs and their applicants may benefit from standardization of program websites and an increase in recruitment and education content.


How neighborhood disorder increases blood pressure in youth: agonistic striving and subordination.
Growing evidence links perceptions of neighborhood disorder to adverse health outcomes but little is known about psychological processes that may mediate this association. We tested the hypothesis that two psychological mechanisms-agonistic striving and subordination-mediate the link between perceived neighborhood disorder and hypertension risk in youth. Perceived neighborhood disorder, agonistic striving, subordination experiences, negative affect, obesity, and ambulatory blood pressure during daily activities (48 h) were assessed in a multiethnic sample of 167 low- to middle-income urban adolescents. Path analyses revealed that agonistic striving, subordination, and obesity each independently mediated the association between neighborhood disorder and blood pressure; these variables accounted for 73 % of the shared variance, 42 % of which was explained by agonistic striving. The direct relationship between perceived neighborhood disorder and blood pressure was no longer significant in the presence of these mediators. Negative affect was associated with neighborhood disorder and subordination, but not blood pressure. Agonistic striving proved to be a significant and substantial mediator of the association between perceived neighborhood disorder, blood pressure, and future hypertension risk. New research should seek to clarify the processes by which stressful neighborhoods induce persistent agonistic motives and perceptions of subordination in adolescents.


Examination of aqueous oxidized cellulose dispersions as a potential drug carrier. II. In vitro and in vivo evaluation of phenylpropanolamine release from microparticles and pellets.
The purpose of this research is to investigate the release of phenylpropanolamine from oxidized cellulose-phenylpropanolamine (OC-PPA) complexes prepared using aqueous OC dispersions (degree of neutralization, DN, 0-0.44) and phenylpropanolamine-hydrochloride (PPA.HCl) (concentration, 0.5 M or 1.4 M) in vitro and in vivo. The results showed a faster drug release from the OC-PPA complex made using the OC dispersion with a DN value of 0.22 than from those prepared using dispersions with DN values of 0.29 to 0.44. No significant difference existed between the release profiles of OC-PPA microparticles made using OC dispersions with DN values of 0.29 to 0.44. OC-PPA complexes that contained smaller size particles or higher drug levels, or that were processed by freeze drying released PPA faster. Compared with microparticles, the pellets of OC-PPA complexes released PPA more slowly initially. An increase in pH or ionic strength of the dissolution medium increased the release of PPA, which is attributable to increased polymer hydration and solubilization at higher pH and ionic strength conditions. The OC-PPA pellets implanted subcutaneously in rats released 100% of their PPA in 9 to 12 hours. A good correlation was found between the in vivo and in vitro release data. Tissue pathology results showed no significant inflammatory tissue reactions. In conclusion, the partially ionized aqueous OC dispersions have the potential to be used as an implantable biodegradable carrier for amine drugs.


Assessing spatial impacts of historical pulp mill effluent on trophic dynamics in a coastal marine ecosystem using stable isotope (δ13C and δ15N) analysis.
Boat Harbour, Nova Scotia was a tidal estuary that was converted into a wastewater treatment facility for pulp mill effluent in 1967. Treated effluent from Boat Harbour was discharged into the coastal Northumberland Strait, contributing significant nutrient and freshwater inputs into the coastal environment, potentially impacting local biogeochemistry and ecosystem structure. This study used stable isotope analysis of carbon (δ13C) and nitrogen (δ15N) of representative taxa to assess spatial variability in nutrient sources and trophic dynamics. Results identified stable isotope variation with depleted δ13C and δ15N values in taxa near Boat Harbour. Blue mussel (Mytilus edulis) and mummichog (Fundulus heteroclitus) were the most suitable bioindicators for identifying variation in nutrient sources. Stable isotope signatures in this study may be reflective of residual pulp mill effluent-derived nutrients, differences in marine versus terrestrial nutrient sources, and a pronounced coastal salinity gradient. The present study defined the baseline nutrient conditions of the Northumberland Strait and will be useful in assessing the effectiveness of remediation activities.


Case Report: Composite pheochromocytoma with ganglioneuroma component: A report of three cases.
Composite pheochromocytoma (CP) is a very rare tumor originating from neural crest cells, predominantly composed of pheochromocytoma (PCC), a chromaffin cell tumor arising in adrenal medulla, and ganglioneuroma, a tumor derived from autonomic ganglion cells of the nervous system. Moreover, CP may be present in the hereditary syndromes of which pheochromocytoma is part. Literature offers scarce data on this subject, and particularly about its biological behavior, clinical evolution, and molecular profile. We report the phenotype and outcome of three cases of CP (PCC and ganglioneuroma components), followed up at the Endocrine Service of the Clementino Fraga Filho University Hospital, Federal University of Rio de Janeiro, UFRJ, Rio de Janeiro, Brazil. Two nonsyndromic patients (cases 1 and 2) were negative to germline mutations in genes VHL, SDHB, SDHC, SDHD, SDHAF2, TMEM127, and MAX, while the third case (case 3) had clinical diagnosis of neurofibromatosis syndrome. Cases 1, 2, and 3 were diagnosed at 29, 39, and 47 years old, respectively, and were followed up for 3, 17, and 9 years without no CP recurrence. All cases had apparent symptoms of catecholaminergic excess secreted by PCC. Ganglioneuroma, the neurogenic component present in all three cases, had a percentage representation ranging from 5% to 15%. Tumors were unilateral and large, measuring 7.0 cm × 6.0 cm × 6.0 cm, 6.0 cm × 4.0 cm × 3.2 cm, and 7.5 cm × 6.0 cm × 4.5 cm, respectively. All cases underwent adrenalectomy with no recurrence, metastasis, or development of contralateral tumor during follow-up. Genetic testing has been scarcely offered to CP cases. However, a similar frequency of genetic background is found when compared with classic PCC, mainly by the overrepresentation of NF1 cases in the CP subset. By literature review, we identified a notorious increase in cases reported with CP in the last decade, especially in the last 3 years, indicating a recent improvement in the diagnosis of this rare disorder in clinical practice.


[Stigma and discrimination: the experiences of HIV-positive women in poor neighborhoods of Maputo, Mozambique].
The HIV/AIDS epidemic is a serious public health problem in Mozambique. The country has high prevalence rates, and the epidemic's impact is aggravated by the stigma affecting HIV-positive persons. This study takes a socio-anthropological perspective to analyze the experience of HIV-positive women in poor neighborhoods of Maputo and the ways they cope with stigma and discrimination. Semi-structured interviews were conducted with 10 HIV-positive women. The results show how gender inequalities increase women's vulnerability to HIV and contribute to their stigmatization and discrimination. In dealing with stigma, women try to keep their diagnosis confidential, seeking support in group meetings with others living with HIV. Public policies should focus on women's empowerment and the reduction of HIV/AIDS-related stigma.


The functions of SID1 transmembrane family, member 2 (Sidt2).
The SID1 transmembrane family, member 2, namely, Sidt2, is a highly glycosylated multichannel lysosomal transmembrane protein, but its specific physiological function remains unknown. Lysosomal membrane proteins are very important for the executive functioning of lysosomes. As an important part of the lysosomal membrane, Sidt2 can maintain the normal morphology of lysosomes and help stabilize them from the acidic pH environment within. As a receptor/transporter, it binds and transports nucleic acids and mediates the uptake and degradation of RNA and DNA by the lysosome. During glucose metabolism, deletion of Sidt2 can cause an increase in fasting blood glucose and the impairment of grape tolerance, which is closely related to the secretion of insulin. During lipid metabolism, the loss of Sidt2 can cause hepatic steatosis and lipid metabolism disorders and can also play a role in signal regulation and transport. Here, we review the function of the lysosomal membrane protein Sidt2, and focus on its role in glucose and lipid metabolism, autophagy and nucleotide (DNA/RNA) transport.


Standardization of Berberis aristata DC and Nigella sativa L. Using HPTLC and GCMS and Their Antineoplasia Activity in 7,12-Dimethylbenz[a]anthracene-Induced Mouse Models.
Berberis aristata DC and Nigella sativa L. are officially listed in various Indian Pharmacopoeia and AYUSH official documents. Prescribed for different ailments for proven medicinal activities, they thus became part of polyherbal medications. With reverse pharmacology and scientific validation, more than 30 patents are filed on different formulations of B. aristata and granted. Nigella sativa L. has been broadly studied for its therapeutic potential and wide range of activities against cardiovascular, diabetic, cancer, and life style disorders. Thus, this study is aimed at standardizing B. aristata and N. sativa and their antineoplasia activity in 7, 12-dimethylbenz[a]anthracene (DMBA)-induced mouse models. Molecular docking was done using the Schrodinger program Maestro 9.0. Herbal extracts and essential oil (B. aristata and N. sativa) were standardized and quantified using high-performance thin-layer chromatography (HPTLC) (CAMAG) and gas chromatography-mass spectrometry (GCMS) (Agilent 2010GC System) with validated methods. DMBA was administered orally once a week (1mg/200 µL) to each animal except the normal control. Hematology, histopathology, and immunoassays were performed, and data were analyzed and depicted with GraphPad and SPSS. In molecular docking, thymoquinone showed the highest docking score (9.519, 9.211, and 9.042, respectively) in the active site pockets of IL6 (PDB ID: 4CNI and 5FCU), TNF (PDB ID: 2AZ5), and VEGF (PDB ID: 4KZN). Out of all four target sites, thymoquinone and berberine showed good binding affinity with IL6 (PDB ID: 4CNI) compared to α- and β-pinenes. HPTLC analysis of the hydroalcoholic extract showed the presence of berberine both qualitatively and quantitatively (5.4% berberine), and thymoquinone detected 0.17% in the N. sativa extract. GCMS for essential oil showed 26 compounds including ±pinene. Leukocytes and erythrocytes of N. sativa and B. aristata were analyzed, and significant improvements were recorded (P < 0.05) and graphically presented. Mean survival time was calculated by the Kaplan Meier method (119 days). Immunoassay analyses were conducted, namely, TNF-α and VEGF, and interpreted and marked.


[Relationship between the content of some fractions of high molecular weight polyphosphates and total nucleic acids upon dehydration of the yeast Saccharomyces cerevisiae].
Essential redistribution of various polyphosphate fractions was shown during dehydration and subsequent reactivation of Saccharomyces cerevisiae 14. Dehydration no matter what method was used, was followed by an increase in the content of acid soluble polyphosphates (fraction Poly P1) and a decrease of that of salt soluble polyphosphates (fraction Poly P2). Reactivation of dehydrated yeast was, on the contrary, accompanied by a decrease in the PP 1 and an increase in the Poly P2 content. A direct correlation between the Poly P2 fraction and total nucleic acids was demonstrated under various conditions of dehydration and subsequent reactivation. An inverse correlation between the content of the Poly P2, fraction and nucleic acids, on the one hand, and that of the Poly P1 fraction, on the other, was observed. Study of activities of polyphosphatases, tripolyphosphatase, pyrophosphatase and ATPase in dehydrated yeast showed values similar to those in original cells.


Chronic ethanol consumption decreases murine Langerhans cell numbers and delays migration of Langerhans cells as well as dermal dendritic cells.
Chronic alcoholics experience increased incidence and severity of infections, the mechanism of which is incompletely understood. Dendritic cells (DC) migrate from peripheral locations to lymph nodes (LN) to initiate adaptive immunity against infection. Little is known about how chronic alcohol exposure affects skin DC numbers or migration. Mice received 20% EtOH in the drinking water for up to 35 weeks. Baseline Langerhans cell (LC) and dermal DC (dDC) numbers were enumerated by immunofluorescence (IF). LC repopulation after inflammation was determined following congenic bone marrow (BM) transplant and ultraviolet (UV) irradiation. Net LC loss from epidermis was determined by IF following TNF-alpha or CpG stimulation. LC and dDC migration into LN was assessed by flow cytometry following epicutaneous FITC administration. Chronic EtOH consumption caused a baseline reduction in LC but not dDC numbers. The deficit was not corrected following transplantation with non-EtOH-exposed BM and UV irradiation, supporting the hypothesis that the defect is intrinsic to the skin environment rather than LC precursors. Net loss of LC from epidermis following inflammation was greatly reduced in EtOH-fed mice versus controls. Ethanol consumption for at least 4 weeks led to delayed LC migration into LN, and consumption for at least 8 weeks led to delayed dDC migration into LN following epicutaneous FITC application. Chronic EtOH consumption causes decreased density of epidermal LC, which likely results in decreased epidermal immunosurveillance. It also results in altered migratory responsiveness and delayed LC and dDC migration into LN, which likely delays activation of adaptive immunity. Decreased LC density at baseline appears to be the result of an alteration in the skin environment rather than an intrinsic LC defect. These findings provide novel mechanisms to at least partially explain why chronic alcoholics are more susceptible to infections, especially those following skin penetration.


Multicenter trial analysis. Fluocinonide and betamethasone gel in psoriasis.
The relative efficacy of fluocinonide and betamethasone benzoate gels in treating the symptoms of psoriasis was evaluated in 335 patients, of whom 263 were analyzable. By means of a paired comparison design in which both drugs were examined simultaneously in the same patient, many potential sources of variation were eliminated. In addition, the analysis took into account the difficulties of pooling results in a multicenter trial. Imposing a rigorous grouping structure on the results from individual clinics minimized differences between clinics and demonstrated consistently high, statistically significant results in favor of fluocinodine gel even when several different statistical approaches were used. Local adverse reactions related to drugs were experienced by few patients.


Effect of duration and frequency of breast-feeding on postpartum amenorrhea.
The relationship among duration and frequency of breast-feeding, age, level of education, and parity on postpartum amenorrhea were investigated retrospectively in selected married Nigerian women attending the two local maternity centers in Ile-Ife, Nigeria, between January and May 1980. The mean values for duration and frequency of breast-feeding per day were 16.48 +/- 5.43 (SD) months and 4.43 +/- 1.46 (SD), respectively. The mean length of postpartum amenorrhea was 12.54 +/- 6.97 (SD) months. Duration and frequency of breast-feeding were positively related to length of postpartum amenorrhea, whereas parity and years of maternal education were negatively related to it. Duration and frequency of breast-feeding were statistically significant (P less then .05), whereas maternal level of education, parity, and age were not. The implications of the findings for pediatricians and other health workers who may wish to promote breast-feeding among their nursing clients are highlighted.


Intake of medication and vitamin status in the elderly.
An inadequate vitamin status is associated with higher morbidity and frailty in the elderly and might be due to medication. This study aimed to evaluate the status of several vitamins in relation to regular intake of medication in this population. A total of 102 non-institutionalized subjects aged 70-90 years were recruited. Plasma levels of vitamins A, D, E, K and C were determined by HPLC. The functional parameters of vitamins B(1), B(2) and B(6), i.e. the activities of the erythrocyte enzymes transketolase, glutathione reductase and glutamic oxaloacetic transaminase were analyzed photometrically; plasma folate and vitamin B(12) were determined by RIA. The status of vitamins A, E and C was generally satisfactory. Eighty-eight percent and 42% of participants were deficient in vitamins D and K, respectively, as were 29% in B(6); up to 10% were deficient in vitamins B(1), B(2), B(12) and folate. A considerable percentage of participants was, however, at risk for vitamin deficiencies (vitamins B(1), B(6), B(12) and folate: 20-30%; vitamin B(2): 60%). Regular intake of maximally 2 drugs per day was not adversely related to the status of several vitamins; intake of ≥ 3 drugs per day was significantly negatively associated with the status of vitamins D, K, B(6) and folate. Daily intake of ≥ 3 drugs was found to be adversely associated with the status of some vitamins in the elderly. Hence, the medication schedule and nutritional status of these subjects should be monitored closely to ensure that the daily micronutrient requirement is fulfilled.


Structural insights into photosystem II supercomplex and trimeric FCP antennae of a centric diatom Cyclotella meneghiniana.
Diatoms are dominant marine algae and contribute around a quarter of global primary productivity, the success of which is largely attributed to their photosynthetic capacity aided by specific fucoxanthin chlorophyll-binding proteins (FCPs) to enhance the blue-green light absorption under water. We purified a photosystem II (PSII)-FCPII supercomplex and a trimeric FCP from Cyclotella meneghiniana (Cm) and solved their structures by cryo-electron microscopy (cryo-EM). The structures reveal detailed organizations of monomeric, dimeric and trimeric FCP antennae, as well as distinct assemblies of Lhcx6_1 and dimeric FCPII-H in PSII core. Each Cm-PSII-FCPII monomer contains an Lhcx6_1, an FCP heterodimer and other three FCP monomers, which form an efficient pigment network for harvesting energy. More diadinoxanthins and diatoxanthins are found in FCPs, which may function to quench excess energy. The trimeric FCP contains more chlorophylls c and fucoxanthins. These diversified FCPs and PSII-FCPII provide a structural basis for efficient light energy harvesting, transfer, and dissipation in C. meneghiniana.


Construct validity of the AIMS-2 upper limb function scales as a measure of disability in individuals with osteoarthritis of the hand.
This study aimed to validate the use of the Arthritis Impact Measurement Scales 2 (AIMS-2) Hand and Finger Function and Arm Function scales for assessing disability in persons with osteoarthritis (OA) of the hand. Ninety-nine subjects with hand OA completed the AIMS-2 scales, grip strength testing, and timed manipulation tasks. Confirmatory factor analyses determined that the ten items comprising the scales separated out into hand and arm limitations. Spearman correlations revealed the appropriate associations between Hand and Finger Function scale scores and grip strength (r = -0.37) and dexterity (r = 0.36). Arm Function scale scores discriminated individuals with additional upper limb involvement from those without (Mann-Whitney test, p < 0.01). Thus, convergent construct validity of the Hand and Finger Function scale and known-group validity of the Arm Function scale were shown. These AIMS-2 scales are valid and distinct measures of disability in individuals with hand OA.


Z-DNA as a Tool for Nuclease-Free DNA Methyltransferase Assay.
Methylcytosines in mammalian genomes are the main epigenetic molecular codes that switch off the repertoire of genes in cell-type and cell-stage dependent manners. DNA methyltransferases (DMT) are dedicated to managing the status of cytosine methylation. DNA methylation is not only critical in normal development, but it is also implicated in cancers, degeneration, and senescence. Thus, the chemicals to control DMT have been suggested as anticancer drugs by reprogramming the gene expression profile in malignant cells. Here, we report a new optical technique to characterize the activity of DMT and the effect of inhibitors, utilizing the methylation-sensitive B-Z transition of DNA without bisulfite conversion, methylation-sensing proteins, and polymerase chain reaction amplification. With the high sensitivity of single-molecule FRET, this method detects the event of DNA methylation in a single DNA molecule and circumvents the need for amplification steps, permitting direct interpretation. This method also responds to hemi-methylated DNA. Dispensing with methylation-sensitive nucleases, this method preserves the molecular integrity and methylation state of target molecules. Sparing methylation-sensing nucleases and antibodies helps to avoid errors introduced by the antibody's incomplete specificity or variable activity of nucleases. With this new method, we demonstrated the inhibitory effect of several natural bio-active compounds on DMT. All taken together, our method offers quantitative assays for DMT and DMT-related anticancer drugs.


Circulating CD2+ monocytes are dendritic cells.
Low levels of CD2 have been described on subsets of monocytes, macrophages, and dendritic cells. CD2 is expressed on about one-third of circulating monocytes, at levels one-half log lower than on T or NK cells, representing 2-4% of PBMC. FACS analysis of CD2+ and CD2- monocytes revealed no significant difference in the expression of adhesion molecules (CD11a/b/c), class II Ags (HLA-DR, -DQ, -DP), myeloid Ags (CD13, CD14, CD33), or costimulatory molecules (CD80, CD86). Freshly isolated CD2+ and CD2- monocytes were morphologically indistinguishable by phase contrast microscopy. However, scanning electron microscopy revealed large prominent ruffles on CD2+ monocytes in contrast to small knob-like projections on CD2- monocytes. After 2 days of culture, the CD2+ monocytes largely lost CD14 expression and developed distinct dendrites, whereas the CD2- monocytes retained surface CD14 and remained round or oval. Freshly isolated CD2+ monocytes were more potent inducers of the allogeneic MLR and more efficiently induced proliferation of naive T cells in the presence of HIV-1 gp120 than did CD2- monocytes. After culture in the presence of GM/CSF and IL-4, CD2+ monocytes were up to 40-fold more potent than monocyte-derived dendritic cells or CD2- monocytes at inducing allogeneic T cell proliferation. These findings suggest that circulating CD2+ and CD2- monocytes are dendritic cells and the precursors of macrophages, respectively. Thus, dendritic cells are far more abundant in the blood than previously thought, and they and precursors of macrophages exist in the circulation as phenotypically, morphologically, and functionally distinct monocyte populations.


Sensorimotor Cortex Reorganization in Alzheimer's Disease and Metal Dysfunction: A MEG Study.
Objective. To verify whether systemic biometals dysfunctions affect neurotransmission in living Alzheimer's disease (AD) patients. Methods. We performed a case-control study using magnetoencephalography to detect sensorimotor fields of AD patients, at rest and during median nerve stimulation. We analyzed position and amount of neurons synchronously activated by the stimulation in both hemispheres to investigate the capability of the primary somatosensory cortex to reorganize its circuitry disrupted by the disease. We also assessed systemic levels of copper, ceruloplasmin, non-Cp copper (i.e., copper not bound to ceruloplasmin), peroxides, transferrin, and total antioxidant capacity. Results. Patients' sensorimotor generators appeared spatially shifted, despite no change of latency and strength, while spontaneous activity sources appeared unchanged. Neuronal reorganization was greater in moderately ill patients, while delta activity increased in severe patients. Non-Cp copper was the only biological variable appearing to be associated with patient sensorimotor transmission. Conclusions. Our data strengthen the notion that non-Cp copper, not copper in general, affects neuronal activity in AD. Significance. High plasticity in the disease early stages in regions controlling more commonly used body parts strengthens the notion that physical and cognitive activities are protective factors against progression of dementia.


Interpolation of missing data in image sequences.
This paper presents a number of model based interpolation schemes tailored to the problem of interpolating missing regions in image sequences. These missing regions may be of arbitrary size and of random, but known, location. This problem occurs regularly with archived film material. The film is abraded or obscured in patches, giving rise to bright and dark flashes, known as "dirt and sparkle" in the motion picture industry. Both 3-D autoregressive models and 3-D Markov random fields are considered in the formulation of the different reconstruction processes. The models act along motion directions estimated using a multiresolution block matching scheme. It is possible to address this sort of impulsive noise suppression problem with median filters, and comparisons with earlier work using multilevel median filters are performed. These comparisons demonstrate the higher reconstruction fidelity of the new interpolators.


Sand bedding as a reservoir for Lactococcus garvieae dissemination in dairy farms.
Lactococcus garvieae is now recognized as a species with clinical significance for human and veterinary medicine. The aim of this study was to evaluate the presence of this pathogen in sand bedding and milk samples. Two farms in Minnesota with problems of clinical and subclinical mastitis due to streptococci-like organisms were selected. Twenty-four Lactococcus garvieae isolates from sand bedding and 18 isolates from quarter milk were comparatively studied using a genotypic approach. RAPD (random amplification of polymorphic DNA) PCR and REP (repetitive element palindromic) PCR experiments highlighted a similar electrophoretic profile. When genes belonging to the core genome of L. garvieae were tested through a MLRT (multilocus restriction typing), we again observed that all L. garvieae isolates coming from sand bedding and milk shared a common profile, distinguishable from previously studied representative L. garvieae strains. These data indicate that the L. garvieae isolated from sand bedding and milk originated from a few strains adapted to persist in the same habitat. This supports the hypothesis that sand bedding can represent a reservoir of L. garvieae strains and be a potential vehicle for their dissemination in dairy farms.


Temporal order judgement for auditory and visual stimuli.
The ability to perceive temporal order for pairs of auditory and visual stimuli was investigated in 12 volunteers. They were asked to make judgements about the order of presented stimuli by pressing two response buttons in a certain order. The performance on auditory and visual tests was studied in relation to the Inter-Stimulus-Interval (ISI), which varied from 5 to 500 ms. In general, the level of performance was similar for the two modalities and the criterion of 75% of correct responses was reached at ISI longer than 40 ms, independently of the modality. These findings are consistent with previous research. However, at ISI of 5 ms, a significantly higher level of correctness was observed for the auditory than visual task. Such a tendency was also observed for ISIs of 10, 20 and 40 ms. Better processing in the auditory compared to the visual task at shorter ISIs may result from a different kind of transduction mechanism at the level of receptive cells in each modality. Alternatively, subjects may use two different kinds of response strategies in the auditory modality, only one being comparable to the response strategy in the visual modality.


A glimpse at locally advanced melanomas in the settings of poverty: pathologist's viewpoint.
Cutaneous melanoma is the most aggressive cancer of the skin arising from pigment-producing cells, known as melanocytes. It is notorious for spreading early to distant locations. Survival of patients with melanoma largely depends on the thickness of the lesion at the primary site thus spotting it early is crucial. Early diagnosis of melanoma, with an improved quality of life and treatment outcomes, is being achieved in some developed nations through screening and health education. On the contrary, as practicing pathologists in a resource-scarce country, we frequently encounter patients with locally advanced melanoma manifesting as ulceration, bleeding, fungation, and bone erosion. Several factors, including low socioeconomic status, medical mistrust, inaccessibility of health facilities, and absent screening and surveillance services can be attributed to the delayed diagnosis. Therefore to alleviate the burden and complications caused by the late presentation of cutaneous melanoma, an urgent massive community mobilization, information campaigning, and the provision of accessible basic primary health care are urgently needed.


How Do Obstetric and Neonatology Teams Communicate Prior to High-Risk Deliveries?
Improving communication in healthcare improves the quality of care and patient outcomes, but communication between obstetric and neonatal teams before and during a high-risk delivery is poorly studied. We developed a survey to study communication between obstetric and neonatal teams around the time of a high-risk delivery. We surveyed neonatologists from North America and asked them to answer questions about their institutions' communication practices. The survey answers revealed variations in communication practices between responders. Most institutions relied on nursing to communicate obstetric information to the neonatal team. Although a minority of institutions used a standardized communication process to summon neonatology team or to communicate in the delivery room, these reported higher rates of information sharing and greater satisfaction with communication between services. Standardized communication procedures are an underutilized method of communication and have the potential to improve communication around high-risk deliveries.


Interaction of retinoic acid radical cation with lysozyme and antioxidants: laser flash photolysis study in microemulsion.
All-trans retinoic acid (ATRA) plays essential roles in the normal biological processes and the treatment of cancer and skin diseases. Considering its photosensitive property, many studies have been focused on the photochemistry of ATRA. In this study, we investigated the transient phenomena in the laser flash photolysis (LFP) of ATRA in microemulsion to further understand the photochemistry of ATRA. Results show that 355 nm LFP of ATRA in both acidic and alkaline conditions leads to the generation of retinoic acid cation radicals (ATRA(•+)) via biphotonic processes. The employment of microemulsion system allows us to investigate the reaction of hydrophobic ATRA(•+) with molecules of different polarity. Therefore, we studied the reaction activity of ATRA(•+) to many hydrophobic and hydrophilic molecules. Results show that ATRA(•+) can efficiently interact with lysozyme, tyrosine, tryptophan and many antioxidants, such as curcumin (Cur), vitamin C (VC) and gallic acid (GA). The apparent rate constants of these reactions were measured and compared. These findings suggest that ATRA(•+) is a reactive transient product which may pose damage to lysozyme, and antioxidants, such as Cur, VC and GA, may inactivate ATRA(•+) by efficient quenching reactions.


Reviewing the ketamine model for schizophrenia.
The observation that antagonists of the N-methyl-D-aspartate receptor (NMDAR), such as phencyclidine (PCP) and ketamine, transiently induce symptoms of acute schizophrenia had led to a paradigm shift from dopaminergic to glutamatergic dysfunction in pharmacological models of schizophrenia. The glutamate hypothesis can explain negative and cognitive symptoms of schizophrenia better than the dopamine hypothesis, and has the potential to explain dopamine dysfunction itself. The pharmacological and psychomimetic effects of ketamine, which is safer for human subjects than phencyclidine, are herein reviewed. Ketamine binds to a variety of receptors, but principally acts at the NMDAR, and convergent genetic and molecular evidence point to NMDAR hypofunction in schizophrenia. Furthermore, NMDAR hypofunction can explain connectional and oscillatory abnormalities in schizophrenia in terms of both weakened excitation of inhibitory γ-aminobutyric acidergic (GABAergic) interneurons that synchronize cortical networks and disinhibition of principal cells. Individuals with prenatal NMDAR aberrations might experience the onset of schizophrenia towards the completion of synaptic pruning in adolescence, when network connectivity drops below a critical value. We conclude that ketamine challenge is useful for studying the positive, negative, and cognitive symptoms, dopaminergic and GABAergic dysfunction, age of onset, functional dysconnectivity, and abnormal cortical oscillations observed in acute schizophrenia.


Parasites and travellers' diarrhoea.
Giardia lamblia is discussed as the most important parasitic cause of travellers' diarrhoea. The parasitology of this flagellate protozoan is discussed with respect to route of infection, threshold infecting dose, and incubation period. The geographic distribution of giardiasis is widespread, being common in tropical and sub-tropical regions and endemic at a low level in European and North American countries. The predisposing factors are detailed together with clinical manifestations, pathogenesis and diagnosis. Effective therapy may be quinacrine t.i.d. for 5-10 days, metronidazole 200 mg t.i.d. for 14 days or 2 g once daily over 3 days, or a single dose of 1.5 g of tinidazole. Although Entamoeba histolytica is distributed world-wide, it rarely causes travellers' diarrhoea. However, the clinical features, diagnosis and treatment of amoebiasis-related travellers' diarrhoea are detailed.


Regulation of immunity to the two-component lantibiotic, lacticin 3147, by the transcriptional repressor LtnR.
Lacticin 3147 is a membrane-active, two-component lantibiotic produced by Lactococcus lactis ssp. lactis DPC3147. In this study, the promoters of the lacticin 3147 gene cluster were mapped to the intergenic region between ltnR and ltnA1 (the genes encoding the regulatory protein LtnR and the first structural gene, LtnA1), and Northern analyses revealed that the biosynthetic and immunity genes are divergently transcribed in two operons, ltnA1A2M1TM2D and ltnRIFE respectively. Although the promoter controlling biosynthesis (Pbac) appears to be constitutive, characterization of a downstream beta-galactosidase (beta-gal) fusion beyond an intragenic stem-loop structure in ltnM1 confirmed that this putative transcriptional attenuator allows limited readthrough to the downstream biosynthetic genes, thus maintaining the correct stoichiometry between structural peptides and biosynthetic machinery. The promoter of the ltnRIFE operon (Pimm) was shown to be regulated by the transcriptional repressor LtnR. A mutant with a truncated ltnR gene exhibited a hyperimmune phenotype, whereas overexpression of ltnR resulted in cells with increased sensitivity to lacticin 3147. Gel mobility shift analysis indicated that LtnR binds to the Pimm promoter region, and fusion of this promoter to the beta-gal gene of pAK80 revealed that expression from Pimm is significantly reduced in the presence of LtnR. Thus, we have demonstrated that lacticin 3147 uses a regulatory mechanism not previously identified in lantibiotic systems.


Pulse modulated 900 MHz radiation induces hypothyroidism and apoptosis in thyroid cells: a light, electron microscopy and immunohistochemical study.
In the present study we investigated the possible histopathological effects of pulse modulated Radiofrequency (RF) fields on the thyroid gland using light microscopy, electron microscopy and immunohistochemical methods. Two months old male Wistar rats were exposed to a 900 MHz pulse-modulated RF radiation at a specific absorption rate (SAR) of 1.35 Watt/kg for 20 min/day for three weeks. The RF signals were pulse modulated by rectangular pulses with a repetition frequency of 217 Hz and a duty cycle of 1:8 (pulse width 0.576 ms). To assess thyroid endocrine disruption and estimate the degree of the pathology of the gland, we analysed structural alterations in follicular and colloidal diameters and areas, colloid content of the follicles, and height of the follicular epithelium. Apoptosis was confirmed by Transmission Electron Microscopy and assessing the activites of an initiator (caspase-9) and an effector (caspase-3) caspases that are important markers of cells undergoing apoptosis. Morphological analyses revealed hypothyrophy of the gland in the 900 MHz RF exposure group. The results indicated that thyroid hormone secretion was inhibited by the RF radiation. In addition, we also observed formation of apoptotic bodies and increased caspase-3 and caspase-9 activities in thyroid cells of the rats that were exposed to modulated RF fields. The overall findings indicated that whole body exposure to pulse-modulated RF radiation that is similar to that emitted by global system for mobile communications (GSM) mobile phones can cause pathological changes in the thyroid gland by altering the gland structure and enhancing caspase-dependent pathways of apoptosis.


Penetration of Candida biofilms by antifungal agents.
A filter disk assay was used to investigate the penetration of antifungal agents through biofilms containing single and mixed-species biofilms containing Candida. Fluconazole permeated all single-species Candida biofilms more rapidly than flucytosine. The rates of diffusion of either drug through biofilms of three strains of Candida albicans were similar. However, the rates of drug diffusion through biofilms of C. glabrata or C. krusei were faster than those through biofilms of C. parapsilosis or C. tropicalis. In all cases, after 3 to 6 h the drug concentration at the distal edge of the biofilm was very high (many times the MIC). Nevertheless, drug penetration failed to produce complete killing of biofilm cells. These results indicate that poor antifungal penetration is not a major drug resistance mechanism for Candida biofilms. The abilities of flucytosine, fluconazole, amphotericin B, and voriconazole to penetrate mixed-species biofilms containing C. albicans and Staphylococcus epidermidis (a slime-producing wild-type strain, RP62A, and a slime-negative mutant, M7) were also investigated. All four antifungal agents diffused very slowly through these mixed-species biofilms. In most cases, diffusion was slower with biofilms containing S. epidermidis RP62A, but amphotericin B penetrated biofilms containing the M7 mutant more slowly. However, the drug concentrations reaching the distal edges of the biofilms always substantially exceeded the MIC. Thus, although the presence of bacteria and bacterial matrix material undoubtedly retarded the diffusion of the antifungal agents, poor penetration does not account for the drug resistance of Candida biofilm cells, even in these mixed-species biofilms.


Physics of surgery with lasers.
The characteristics of the principal lasers used in surgery are summarized in Table 1. Their diverse effects on biologic tissues permit the following generalizations: The CO2 laser is best suited for precise, visually controllable tissue removal by vaporization with minimal marginal damage. Hemostasis is excellent for bleeding from capillary vessels, but difficult for larger ones. The Nd:YAG laser is best suited for the coagulation of larger tissue volumes of the order of 10 mm3 or more. Tissue heating inherently extends for several millimeters, leading to excellent hemostasis. Radiation from this laser is well transmitted through flexible optical fibers and clear fluids. The argon ion laser emits radiation in the visible range and is ideally suited for treating the retina and other tissues in the eye without damage to its transparent structures. Radiation of this laser is strongly absorbed by pigmented tissues, scattered and reflected by others, and transmitted by fluids. Its radiation can be focused to very small spot sizes, leading to high precision and high-power densities. It has hemostatic properties intermediate between those of the CO2 and of the Nd:YAG laser radiations. It is well transmitted through optical fibers and clear fluids. It is used extensively in ophthalmology and dermatology. Selected applications to neurosurgery and otology are being investigated. These lasers have become indispensable adjuncts to the surgical armamentarium of several specialties. The very success of these lasers is leading to a critical examination of their shortcomings and to a search for improved systems. Examples are (1) the ongoing search for optical fibers to transmit the radiation of the CO2 laser; (2) the development of systems for the sequential delivery to tissues of several wavelengths from a single unit (Fig. 14); and (3) investigations of tissue effects of laser beams in the ultraviolet and in the infrared at wavelengths intermediate between those of the Nd:YAG and CO2 lasers. The use of lasers has already contributed to improved medical care in many surgical disciplines. Additional areas of application can be confidently anticipated.


Comparative study on efficacy of three sperm-separation techniques.
To evaluate the comparative effectiveness of real-time sperm separation technique (Wang's tube method) and other two conventional methods in isolating high-quality sperm preparation, and to compare the spouse pregnancy rate in intrauterine insemination (IUI) with sperm preparations isolated by these methods. The effectiveness of the real-time sperm separation technique, the conventional swim-up and the Percoll discontinuous density gradient methods in isolating sperm preparations from 60 infertile patients (20 with apparently normal semen and 40, abnormal semen contaminated with microorganisms and other impurities) was evaluated and compared. The microorganisms to be removed included bacteria, virus, Chlamydia trachomaticum, Ureaplsama urealyticum, etc. The spouse pregnancy rates in IUI with sperm preparations isolated by these three techniques from 80 oligoasthenoteratospermic patients were also compared. The quality (including the percentages of normal form, normal-chromatin and motile sperm, and the grade of motility) of sperm obtained by the real-time sperm separation technique was much higher (P < 0.01) as compared with those by the other two methods. The Wang's tube method was also more effective in removing microorganisms and other impurities. The method provided a higher IUI pregnancy rate than the other two sperm separation techniques (P < 0.05). The real-time sperm separation technique is the most effective method so far available in isolating high-quality sperm samples to be used in assisted reproduction.