Detection of ERG11 point mutations in Iranian fluconazole-resistant Candida albicans isolates.
Candidiasis is referred to a group of superficial and deep-tissue fungal infections often caused by Candida albicans. The superficial infections affect the oral, oropharynx, esophagus, and vaginal mucosa. The treatment of choice for these infections is the use of azoles, such as fluconazole. However, the increased use of these antifungal agents has led to the emergence of azole-resistant isolates of C. albicans. Different mechanisms have been suggested for the development of drug resistance, such as mutations in the encoding gene ERG11. Mutations in ERG11 result in changes in the ERG11p spatial construction and reduce the affinity between the protein and azole. This study aimed to determine the susceptibility profile of C. albicans clinical isolates to fluconazole using microdilution method. The present research was also targeted toward the detection of mutations that might be related to fluconazole resistance by the amplification and sequencing of ERG11 gene. This study was conducted on a total of 216 clinical isolates obtained from Mashhad, Isfahan, and Tehran cities in Iran, during 2016-2018. The clinical isolates were identified using molecular techniques. Furthermore, minimum inhibitory concentration (MICs) was determined according to the clinical and laboratory standards institute M27-A3 and M27-S4 documents. The concentration range for fluconazole was obtained as 0.063-64 μg/ml. In the resistant strains, ERG11 genes were amplified by specific primers. Subsequently, cycle sequencing reactions were performed on purified polymerase chain reaction (PCR) products in forward and reverse directions. Finally, the results were analyzed by MEGA (version 7) and Gene Runner software (version 6.5.30). Out of 216 strains, 100 (46.3%) species were identified as C. albicans. The MIC values for fluconazole had a range of 0.125-16 μg/ml with the MIC50 and MIC90 values of 0.5 and 1 μg/ml, respectively. Totally, 41 nucleotide changes were detected among 4 resistant isolates. In this regard, 4 out of 41 mutations in codons caused changes in ERG11p; however, these mutations did not lead to fluconazole resistance. Fluconazole resistance among clinical isolates is not merely due to the changes in ERG11p. This resistance may be also related to some other mechanisms, such as the prevention of the intracellular accumulation of the antifungal agent and alteration of the target enzyme to diminish drug binding.


First Experimental Assessment of Protein Intrinsic Disorder Involvement in an RNA Virus Natural Adaptive Process.
Intrinsic disorder (ID) in proteins is defined as a lack of stable structure in physiological conditions. Intrinsically disordered regions (IDRs) are highly abundant in some RNA virus proteomes. Low topological constraints exerted on IDRs are expected to buffer the effect of numerous deleterious mutations and could be related to the remarkable adaptive potential of RNA viruses to overcome resistance of their host. To experimentally test this hypothesis in a natural pathosystem, a set of four variants of Potato virus Y (PVY; Potyvirus genus) containing various ID degrees in the Viral genome-linked (VPg) protein, a key determinant of potyvirus adaptation, was designed. To estimate the ID contribution to the VPg-based PVY adaptation, the adaptive ability of the four PVY variants was monitored in the pepper host (Capsicum annuum) carrying a recessive resistance gene. Intriguingly, the two mutants with the highest ID content displayed a significantly higher ability to restore infection in the resistant host, whereas the less intrinsically disordered mutant was unable to restore infection. The role of ID on virus adaptation may be due either to a larger exploration of evolutionary pathways or the minimization of fitness penalty caused by resistance-breaking mutations. This pioneering study strongly suggests the positive impact of ID in an RNA virus adaptive capacity.


Using plastic bag waste to assess the reliability of self-reported waste disposal data.
Direct waste analysis (DWA) and the material flows approach are the two standard methods to quantify aggregated waste streams and analyze waste composition. Yet, with the increasing application of producer responsibility measures, product-based waste data rather than aggregated waste data are becoming important. It is over this requirement that both approaches fail to some extent in delivering the type and quality of information that is needed. This study uses plastic bag waste as an illustration to show how self-reported questionnaire survey data may be used to assess disposal quantities of product-based waste types. The estimates from a large-scale questionnaire survey with over 4,100 completed cases were verified against DWA data of the same year in Hong Kong. It was found that self-reported data give systematically lower figures (on the order of 1.3-5 times) than those obtained from standard methods such as DWA for Hong Kong and the UK. However, it is demonstrated that self-reported data can be internally consistent. Also, the magnitude of underestimation may not be as considerable as it appears since the data from DWA are not themselves entirely accurate owing to the difficulties in obtaining a pure load of waste for field analysis and the variable moisture contents or contamination levels in waste material.


Lipid content evaluation of Drosophila tumour associated haemocytes through Third Harmonic Generation measurements.
Non-linear microscopy is a powerful imaging tool to examine structural properties and subcellular processes of various biological samples. The competence of Third Harmonic Generation (THG) includes the label free imaging with diffraction-limited resolution and three-dimensional visualization with negligible phototoxicity effects. In this study, THG records and quantifies the lipid content of Drosophila haemocytes, upon encountering normal or tumorigenic neural cells, in correlation with their shape or their state. We show that the lipid accumulations of adult haemocytes are similar before and after encountering normal cells. In contrast, adult haemocytes prior to their interaction with cancer cells have a low lipid index, which increases while they are actively engaged in phagocytosis only to decrease again when haemocytes become exhausted. This dynamic change in the lipid accrual of haemocytes upon encountering tumour cells could potentially be a useful tool to assess the phagocytic capacity or activation state of tumour-associated haemocytes.


Single molecule detection of RNA reporter probes by amplification with Q beta replicase.
The addition of target-specific probe sequences within MDV RNA, an otherwise efficient template for Q beta replicase, generally resulted in RNA molecules with inferior replication properties, including reduced replication rates and poor sensitivities. We have discovered that the replication characteristics of MDV RNA molecules with internally placed probe sequences are dramatically affected by short RNA sequences (spacer elements) at the 5' and 3' ends of the probe sequence. For a given probe sequence, the sequences of the flanking spacer elements effected replication sensitivity by six orders of magnitude and replication rate by three fold. By taking advantage of spacer elements, internal MDV probes were developed that permitted the reproducible, real time, fluorescence detection of a single RNA molecule in less than 25 min through amplification with Q beta replicase. RNA structural analysis of such probes suggested that the spacer elements functioned by allowing the RNA to fold in a way which substantially maintained the tertiary structure of the MDV domain. MDV reporter probes with suitable replication properties were obtained from libraries of RNA molecules in which the probe sequence was flanked by many different spacer elements (generated by random nucleotide synthesis). We demonstrated that this is a general method for developing RNA reporter molecules which are rapidly and reproducibly amplified by Q beta replicase, even from a single molecule.


Anti-contractile effects of perivascular adipose tissue in thoracic aorta from rats fed a high-fat diet: role of aerobic exercise training.
The aim of the present study was to evaluate the effects of aerobic exercise training on perivascular adipose tissue (PVAT) function in thoracic aorta from rats fed a high-fat diet. Aortic vascular reactivity was performed in sedentary (SD), trained (TR), sedentary high-fat diet (SD-HF), and trained high-fat diet (TR-HF) male Wistar rats in the absence (PVAT-) or in the presence (PVAT+) of thoracic PVAT. We also measured circulatory concentrations of leptin and tumour necrosis factor alpha (TNF-α), as well as the protein expressions of TNF-α receptor 1 (TNFR1) and inducible nitric oxide synthase (iNOS) on PVAT. In the SD-HF group, the body weight, epididymal fat pad, thoracic PVAT, circulatory triglycerides, insulin, leptin and TNF-α were increased when compared with the SD group, whereas exercise training reduced these values in TR-HF group. The relaxing response curves to acetylcholine and sodium nitroprusside were not modified by either intervention (high-fat diet or exercise training) or the presence of PVAT. The presence of PVAT had an anti-contractile effect in response to serotonin in all groups. In SD-HF group, the increased magnitude of anti-contractile effects was in parallel with an up-regulation of iNOS protein expression in PVAT without alteration in TNFR1. Exercise training was effective in normalizing the vascular reactivity in rings PVAT+ and in reducing the iNOS protein expression. Exercise training prevented the PVAT-induced alteration in thoracic aorta from rats fed a high-fat diet.


Ellagic Acid Enhances the Antitumor Efficacy of Bevacizumab in an In Vitro Glioblastoma Model.
The anticarcinogenic effect of ellagic acid (EA), a natural phenol of fruits and vegetables, has been investigated in several types of tumors. The combined effect of EA with bevacizumab (BEV), a common drug used in treatment of recurrent glioma, on glioblastoma has not been reported. This study observed the combined effect of EA with BEV on the expression profile of the C6 glioma cell line. Rat C6 glioma cells were treated with EA at 100 μmol/L concentration in combination with BEV at 100 ng/mL concentration for 24, 48, and 72 hours. Cell proliferation was detected by 5-bromo-2'-deoxyuridine immunohistochemistry, and p53 and caspase-3 protein levels were determined by immunohistochemistry and assessed by the H-Score. Expression profiles for P-glycoprotein (MDR1), O6-methylguanine DNA methyltransferase (MGMT), caspase-3, and p53 related proteins were detected by reverse transcriptase polymerase chain reaction after EA treatment with or without BEV. EA combined with BEV conspicuously reduced the cell viability of C6 glioma cells for all incubation times. EA significantly downregulated expression of MGMT regardless of combination with BEV even in the early hours after treatment. Combined EA and BEV reduced MDR1 expression only at 72 hours. EA affected the apoptotic proteins of p53 and caspase-3 at protein level in a time-dependent manner, but not at gene level. This study suggests successful antiproliferative efficacy of EA combined with BEV, probably through inhibition of MGMT expression and time-dependent inhibition of MDR1. EA combined with BEV may be an alternative treatment for drug-resistant gliomas.


Sequences within both the N- and C-terminal domains of phytochrome A are required for PFR ubiquitination and degradation.
Photoconversion of the plant photoreceptor phytochrome A (phyA) from its inactive Pr form to its biologically active Pfr from initiates its rapid proteolysis. Previous kinetic and biochemical studies implicated a role for the ubiquitin/26S proteasome pathway in this breakdown and suggested that multiple domains within the chromoprotein are involved. To further resolve the essential residues, we constructed a series of mutant PHY genes in vitro and analyzed the Pfr-specific degradation of the resulting photoreceptors expressed in transgenic tobacco. One important site is within the C-terminal half of the polypeptide as its removal stabilizes oat phyA as Pfr. Within this half is a set of conserved lysines that are potentially required for ubiquitin attachment. Substitution of these lysines did not prevent ubiquitination or breakdown of Pfr, suggesting either that they are not the attachment sites or that other lysines can be used in their absence. A small domain just proximal to the C-terminus is essential for the form-dependent breakdown of the holoprotein. Removal of just six amino acids in this domain generated a chromoprotein that was not rapidly degraded as Pfr. Using chimeric photoreceptors generated from potato PHYA and PHYB, we found that the N-terminal half of phyA is also required for Pfr-specific breakdown. Only those chimeras containing the N-terminal sequences from phyA were ubiquitinated and rapidly degraded as Pfr. Taken together, our data demonstrate that, whereas an intact C-terminal domain is essential for phyA degradation, the N-terminal domain is responsible for the selective recognition and ubiquitination of Pfr.


Deregulation: causes and consequences.
As a consequence of deregulation, there have been fundamental changes in the way transportation and communications firms are conducting business. Companies are finding that they must be driven by market opportunities and financial needs, not by regulatory considerations. Prices must be based on cost, operations must become more efficient, and consumer-oriented product niches must be found. Moreover, deregulation has added a new element in industrial dynamics, by fostering more relations between the various modes of transportation and by cross-fertilization in computer and telecommunication technology. All in all, a more competitive and innovative spirit emerges from deregulation.


Innate immune defences in the human endometrium.
The human endometrium is an important site of innate immune defence, giving protection against uterine infection. Such protection is critical to successful implantation and pregnancy. Infection is a major cause of preterm birth and can also cause infertility and ectopic pregnancy. Natural anti-microbial peptides are key mediators of the innate immune system. These peptides, between them, have anti-bacterial, anti-fungal and anti-viral activity and are expressed at epithelial surfaces throughout the female genital tract. Two families of natural anti-microbials, the defensins and the whey acidic protein (WAP) motif proteins, appear to be prominent in endometrium. The human endometrial epithelium expresses beta-defensins 1-4 and the WAP motif protein, secretory leukocyte protease inhibitor. Each beta-defensin has a different expression profile in relation to the stage of the menstrual cycle, providing potential protection throughout the cycle. Secretory leukocyte protease inhibitor is expressed during the secretory phase of the cycle and has a range of possible roles including anti-protease and anti-microbial activity as well as having effects on epithelial cell growth. The leukocyte populations in the endometrium are also a source of anti-microbial production. Neutrophils are a particularly rich source of alpha-defensins, lactoferrin, lysozyme and the WAP motif protein, elafin. The presence of neutrophils during menstruation will enhance anti-microbial protection at a time when the epithelial barrier is disrupted. Several other anti-microbials including the natural killer cell product, granulysin, are likely to have a role in endometrium. The sequential production of natural anti-microbial peptides by the endometrium throughout the menstrual cycle and at other sites in the female genital tract will offer protection from many pathogens, including those that are sexually transmitted.


Extrapolated high-order propagators for path integral Monte Carlo simulations.
We present a new class of high-order imaginary time propagators for path integral Monte Carlo simulations that require no higher order derivatives of the potential nor explicit quadratures of Gaussian trajectories. Higher orders are achieved by an extrapolation of the primitive second-order propagator involving subtractions. By requiring all terms of the extrapolated propagator to have the same Gaussian trajectory, the subtraction only affects the potential part of the path integral. The resulting violation of positivity has surprisingly little effects on the accuracy of the algorithms at practical time steps. Thus in principle, arbitrarily high order algorithms can be devised for path integral Monte Carlo simulations. We verified the fourth, sixth, and eighth order convergences of these algorithms by solving for the ground state energy and pair distribution function of liquid (4)He, which is representative of a dense, and strongly interacting, quantum many-body system.


Regulation of alpha-bungarotoxin receptor accumulation in chick retina cultures: effects of membrane depolarization, cyclic nucleotide derivatives, and Ca2+.
The regulation of a putative neuronal nicotinic acetylcholine receptor, the alpha-bungarotoxin (alpha-Btx)-binding protein, was investigated in primary cultures of chick embryo retina. Depolarization of the cells by veratrum alkaloids, or by a high K+ concentration of the culture medium, reduced the accumulation of cell surface alpha-Btx receptors in these cultures. In contrast, the addition of a blocker of voltage-dependent Na+ channels, tetrodotoxin, increased alpha-Btx receptor accumulation in differentiated retina cultures. Derivatives of cyclic adenosine 3':5'-monophosphate also increased the number of alpha-Btx receptor sites, whereas dibutyryl cyclic guanosine 3':5'-monophosphate, but not 8-bromo-cyclic guanosine 3':5'-monophosphate, had the opposite effect. The blocker of voltage-dependent Ca2+ channels, D600, and media containing a reduced Ca2+ concentration also increased alpha-Btx receptor levels. All of these different culture conditions altered the rate of receptor loss after blocking protein synthesis by cycloheximide. These results show that the synthesis of the neuronal alpha-Btx receptor is regulated by membrane depolarization, cyclic nucleotides, and Ca2+ in a fashion analogous to the regulation of muscle acetylcholine receptor.


Role of AVP in malignant DOC-salt hypertension: studies using vascular and antidiuretic antagonists.
To investigate the role of arginine vasopressin (AVP) in the maintenance of blood pressure in deoxycorticosterone (DOC)-salt hypertension, the effects of specific pressor and antidiuretic antagonists of AVP were studied in conscious, freely moving rats with established malignant DOC-salt hypertension. Plasma AVP level was significantly higher in hypertensive than in normotensive animals (4.8 +/- 1.0 vs. 2.0 +/- 0.3 fmol/ml, n = 5, P less than 0.02). Administration of d(CH2)5-d-Leu-VAVP, 10 micrograms/kg, an AVP antagonist that blocked the antidiuretic, but not the pressor effect of exogenous AVP, induced diuresis, and caused a transient fall in blood pressure from 173 +/- 3 to 167 +/- 4 mmHg (n = 8, P less than 0.01) with a concomitant slight increase in heart rate. Similar changes were observed after administration of d(CH2)5Tyr(Et)VAVP, 10 micrograms/kg, an antidiuretic plus pressor antagonist of AVP. Intravenous injection of d(CH2)5Tyr(Me)AVP, 10 micrograms/kg, a specific AVP pressor antagonist had no effect on blood pressure or heart rate, although it completely abolished the pressor response to exogenous AVP. Plasma renin activity remained suppressed following administration of all AVP antagonists. These findings suggest that if AVP should contribute to maintaining high blood pressure in malignant DOC-salt hypertension it would have to be the results of its antidiuretic and not its vasoconstrictor property.


A Cadaver Study to Assess the Feasibility of a Cross-Nerve Transfer of the Infraorbital Nerve for Patients With Peripheral Infraorbital Nerve Injury.
Patients with facial fracture or head and neck surgery sometimes suffer from infraorbital nerve injury. This injury results in severe hemilateral numbness in the midfacial area. The infraorbital nerve ends with two major branches; the infra nasal branch (INB) and superior labial branch (SLB). In this study, we assessed the feasibility of cross-nerve transfer of the INB and SLB based on a cadaver study. The INB/SLB from a total 20 sides of 10 cadavers (2 men and 8 women; average age, 79.9 years) were dissected. The distribution patterns of the INB and SLB, the distance between the INB/SLB and the piriform aperture, and the shortest distance between the INB/SLB were estimated. Three distribution patterns of the INB and SLB were observed, that is type A (65%); only the INB is thick enough for a nerve transfer, type B (20%); only the SLB is thick enough for a nerve transfer, and a combination of types A and B (15%). The distance between the INB, SLB and the piriform aperture was on average 8.61 and 10.81 mm in each. The shortest distance between the INB and SLB was on average 11.34 ± 3.7 mm. The INB and SLB existed in all the specimens and could be found approximately 1 cm below the piriform aperture. The average distance between the INB and SLB was approximately 11 mm. These results imply the feasibility of a cross-nerve transfer of the distal part of the infraorbital nerve.


Long-term exposure to elevated carbon dioxide does not alter activity levels of a coral reef fish in response to predator chemical cues.
Levels of dissolved carbon dioxide (CO2) projected to occur in the world's oceans in the near future have been reported to increase swimming activity and impair predator recognition in coral reef fishes. These behavioral alterations would be expected to have dramatic effects on survival and community dynamics in marine ecosystems in the future. To investigate the universality and replicability of these observations, we used juvenile spiny chromis damselfish (Acanthochromis polyacanthus) to examine the effects of long-term CO2 exposure on routine activity and the behavioral response to the chemical cues of a predator (Cephalopholis urodeta). Commencing at ~3-20 days post-hatch, juvenile damselfish were exposed to present-day CO2 levels (~420 μatm) or to levels forecasted for the year 2100 (~1000 μatm) for 3 months of their development. Thereafter, we assessed routine activity before and after injections of seawater (sham injection, control) or seawater-containing predator chemical cues. There was no effect of CO2 treatment on routine activity levels before or after the injections. All fish decreased their swimming activity following the predator cue injection but not following the sham injection, regardless of CO2 treatment. Our results corroborate findings from a growing number of studies reporting limited or no behavioral responses of fishes to elevated CO2. Alarmingly, it has been reported that levels of dissolved carbon dioxide (CO2) forecasted for the year 2100 cause coral reef fishes to be attracted to the chemical cues of predators. However, most studies have exposed the fish to CO2 for very short periods before behavioral testing. Using long-term acclimation to elevated CO2 and automated tracking software, we found that fish exposed to elevated CO2 showed the same behavioral patterns as control fish exposed to present-day CO2 levels. Specifically, activity levels were the same between groups, and fish acclimated to elevated CO2 decreased their swimming activity to the same degree as control fish when presented with cues from a predator. These findings indicate that behavioral impacts of elevated CO2 levels are not universal in coral reef fishes.


Cytotoxic T cells in chronic idiopathic neutropenia express restricted antigen receptors.
Chronic idiopathic neutropenia (CIN) is an acquired disorder of granulopoiesis characterized by female predominance and mostly uncomplicated course. Crucial to CIN pathophysiology is the presence of activated T lymphocytes with myelosuppressive properties in both peripheral blood (PB) and bone marrow (BM). We systematically profiled the T cell receptor beta chain (TRB) gene repertoire in CD8+ cells of 34 CIN patients through subcloning/Sanger sequencing analysis of TRBV-TRBD-TRBJ gene rearrangements. Remarkable repertoire skewing and oligoclonality were observed, along with shared clonotypes between different patients, alluding to antigen selection. Cross-comparison of our sequence dataset with public TRB sequence databases revealed that CIN may rarely share common immunogenetic features with other entities, however, the CIN TRB repertoire is largely disease-biased. Overall, these findings suggest that CIN may be driven by long-term exposure to a restricted set of specific CIN-associated antigens.


Potential of molecular biophysical stimulation therapy in chronic musculoskeletal disorders: a narrative review.
Current treatment of chronic musculoskeletal diseases does not give sufficient results despite the implementation of novel drugs and techniques in orthopaedics and physical therapy. For instance, osteoporosis treatment is currently mainly limited to drug application, while the goal of osteoarthritis treatment is to mitigate pain symptoms through physical therapy. The main therapeutic principle in the management of osteoporosis is not only to increase bone mass, but also to improve bone and the cartilage quality, which depends on the biomechanical balance. Therefore, there is a strong demand for advanced technologies that would safely and non-invasively accelerate cartilage regeneration and improve bone density. Ten years ago, a new state-of-the-art technology - "Molecular biophysical stimulation therapy (MBST)", specifically nuclear magnetic resonance therapy, emerged on the medical technology market and until now, it has shown successful results in the conservative treatment of musculoskeletal disorders, including back pain. The aim of this review is to provide an integrated, synthesized overview of the current evidence of efficacy of MBST for managing chronic musculoskeletal disorders.


Phylogenetic analysis of simian Plasmodium spp. infecting Anopheles balabacensis Baisas in Sabah, Malaysia.
Anopheles balabacensis of the Leucospyrus group has been confirmed as the primary knowlesi malaria vector in Sabah, Malaysian Borneo for some time now. Presently, knowlesi malaria is the only zoonotic simian malaria in Malaysia with a high prevalence recorded in the states of Sabah and Sarawak. Anopheles spp. were sampled using human landing catch (HLC) method at Paradason village in Kudat district of Sabah. The collected Anopheles were identified morphologically and then subjected to total DNA extraction and polymerase chain reaction (PCR) to detect Plasmodium parasites in the mosquitoes. Identification of Plasmodium spp. was confirmed by sequencing the SSU rRNA gene with species specific primers. MEGA4 software was then used to analyse the SSU rRNA sequences and bulid the phylogenetic tree for inferring the relationship between simian malaria parasites in Sabah. PCR results showed that only 1.61% (23/1,425) of the screened An. balabacensis were infected with one or two of the five simian Plasmodium spp. found in Sabah, viz. Plasmodium coatneyi, P. inui, P. fieldi, P. cynomolgi and P. knowlesi. Sequence analysis of SSU rRNA of Plasmodium isolates showed high percentage of identity within the same Plasmodium sp. group. The phylogenetic tree based on the consensus sequences of P. knowlesi showed 99.7%-100.0% nucleotide identity among the isolates from An. balabacensis, human patients and a long-tailed macaque from the same locality. This is the first study showing high molecular identity between the P. knowlesi isolates from An. balabacensis, human patients and a long-tailed macaque in Sabah. The other common simian Plasmodium spp. found in long-tailed macaques and also detected in An. balabacensis were P. coatneyi, P. inui, P. fieldi and P. cynomolgi. The high percentage identity of nucleotide sequences between the P. knowlesi isolates from the long-tailed macaque, An. balabacensis and human patients suggests a close genetic relationship between the parasites from these hosts.


PEO flocculation with phenolic microparticles.
Polystyrene latex and precipitated calcium carbonate (PCC) with and without dextran sulfate pretreatment were flocculated by the consecutive addition of high-molecular-weight poly(ethylene oxide) and a novel composite latex microparticle consisting of a polystyrene core and a poly(p-vinylphenol) shell. Good flocculation of polystyrene latex and PCC was obtained, whereas the PCC coated with dextran sulfate was not flocculated. The interaction of the composite microparticle with the target colloids was governed by electrostatic forces, whereas hydrogen bonding and hydrophobic interactions drove the PEO adsorption onto the composite particles.


[Brain effects of cannabis--neuroimaging findings].
Cannabis is the most widely used illicit drug. Despite this, only a small number of studies have investigated the long-term neurotoxic consequences of cannabis use. Structural and functional neuroimaging techniques are powerful research tools to investigate possible cannabis-induced pathophysiological changes. A computer literature review was conducted in the MEDLINE and PsycLIT databases between 1966 and November of 2004 with the search terms 'cannabis', 'marijuana', 'neuroimaging', 'magnetic resonance', 'computed tomography', 'positron emission tomography', 'single photon emission computed tomography", 'SPET', 'MRI' and 'CT'. Structural neuroimaging studies have yielded conflicting results. Most studies report no evidence of cerebral atrophy or regional changes in tissue volumes, and one study suggested that long-term users who started regular use on early adolescence have cerebral atrophy as well as reduction in gray matter. However, several methodological shortcomings limit the interpretation of these results. Functional neuroimaging studies have reported increases in neural activity in regions that may be related with cannabis intoxication or mood-change effects (orbital and mesial frontal lobes, insula, and anterior cingulate) and decreases in activity of regions related with cognitive functions impaired during acute intoxication. The important question whether residual neurotoxic effects occur after prolonged and regular use of cannabis remains unclear, with no study addressing this question directly. Better designed neuroimaging studies, combined with cognitive evaluation, may be elucidative on this issue.


Schizophrenia spectrum disorders in later life: prevalence and distribution of age at onset and sex in a dutch catchment area.
The prevalence of schizophrenia in later life is affected by both outflow of early onset patients, due to recovery and excess mortality, and inflow of patients with a later age at onset, making it likely that characteristics of older patients differ markedly from younger patients. We assessed the prevalence of schizophrenia and spectrum disorders and their distribution according to age at onset and sex in an elderly population. Case register study. All patients age 60 years and older, in contact with the Mental Health Organization in a psychiatric catchment area in Amsterdam (the Netherlands), diagnosed with schizophrenia, schizoaffective disorder, or delusional disorder. One-year prevalence estimates, including rates according to age group, age at onset, and sex. In addition, we determined the effect of using different criteria for age at onset. The one-year prevalence of all disorders was 0.71%, subdivided in 0.55% for schizophrenia, 0.14% for schizoaffective disorder, and 0.03% for delusional disorder. The one-year prevalence of early-onset schizophrenia was 0.35%, of late-onset schizophrenia 0.14%, and of very-late-onset schizophrenia-like psychosis 0.05%. Variation of onset criterion affected the proportion of early-onset versus late-onset schizophrenia patients stronger in women than in men. Women outnumbered men markedly in the prevalence estimates for most diagnostic subgroups, including early-onset schizophrenia. We found the prevalence of schizophrenia among older persons to be well within the range reported for younger populations. The considerable proportion with a later age at onset and the strong female preponderance are distinguishing characteristics of older patients with clinical implications.


The bias of isotonic regression.
We study the bias of the isotonic regression estimator. While there is extensive work characterizing the mean squared error of the isotonic regression estimator, relatively little is known about the bias. In this paper, we provide a sharp characterization, proving that the bias scales as O(n -β/3) up to log factors, where 1 ≤ β ≤ 2 is the exponent corresponding to Hölder smoothness of the underlying mean. Importantly, this result only requires a strictly monotone mean and that the noise distribution has subexponential tails, without relying on symmetric noise or other restrictive assumptions.


Preparation and evaluation of piroxicam-HPMCAS solid dispersions for ocular use.
The aim of the study was in vitro evaluation of piroxicam solid dispersions containing hydroxypropyl methylcellulose acetate succinate (HPMCAS-LF, -HF) as a carrier. Binary (piroxicam-HPMCAS) and ternary (piroxicam-HPMCAS-Carbopol 940) solid dispersions were prepared by spray-drying method. The morphological characteristics were investigated by scanning electron microscopy. X-ray diffraction and differential scanning calorimetry were employed to study physical and chemical properties. In vitro release was studied using a flow-through cell technique. Studies of dissolution rate of piroxicam from solid dispersions were carried out in comparison with corresponding physical mixtures and drug alone. The dissolution profiles depend on the presence of Carbopol 940 in solid dispersions.


[Expression of interleukin-21 in the lungs of mice with emphysema and its effects on the differentiation of CD4+ T cell].
To evaluate the expression of interleukin (IL)-21 in a cigarette smoke-induced mice model of emphysema and explore its effects on the differentiation of CD4(+)T cell. Twenty male Balb/c mice were randomly divided into two groups: control group and smoke-exposed group. Morphological changes were evaluated by mean linear intercepts and alveolar destructive index. The proportion of CD4(+)IL-21R(+)T cells in lungs of mice was determined by flow cytometry. And the levels of IL-21 in lungs of mice were analyzed by enzyme-linked immunosorbent assay (ELISA). Fresh lung mononuclear cells were isolated from the smoke-exposed group and divided further into two sub-groups: blank sub-group and co-culture sub-group. Two sub-groups were cultured in medium with or without IL-21 for 24 h and 48 h. The proportions of Th1 and Th17 cells in cell culture medium were determined by flow cytometry. Mean linear intercepts and alveolar destructive index in the smoke-exposed group ((48.6 ± 4.8) µm and 44.9 ± 2.8) were significantly higher than the control group ((32.4 ± 4.0) µm and 28.1 ± 2.1, both P < 0.05). In lungs, the percentage of CD4(+)IL-21R(+)T cells in the smoke-exposed group (4.1% ± 1.5%) significantly increased than that in the control group (1.4% ± 0.4%) (P < 0.05). The levels of IL-21 in lung of the smoke-exposed group ((851 ± 28) ng/L) were higher than those in the control group ((415 ± 39) ng/L, P < 0.05). In lungs, the levels of IL-21 had positive correlations with mean linear intercepts and alveolar destructive index (r = 0.892 and 0.955, both P < 0.05). The percentages of Th1 and Th17 cells in cell culture medium of the co-culture sub-group for 24 h and 48 h significantly increased versus those in the blank group (all P < 0.05). IL-21 may participate in the occurrence and development of emphysema through the induced differentiation of CD4(+)T cells and the promotion of Th1 and Th17-cell responses in lungs.


Streptococcal involvement in childhood acute glomerulonephritis: a review of 20 cases at admission.
Twenty children sequentially admitted to the paediatric ward, Tauranga Hospital, with a diagnosis of acute glomerulonephritis, are reviewed for evidence of recent streptococcal infection as at admission. Sixteen children (80 percent) were shown to have at least one streptococcal antibody titre (ASO and anti-DNase B) elevated above the upper limit of normal for the population. Two children had negligible antibody levels detected. Case histories and/or cultural results provided additional supportive evidence for recent streptococcal infection. Serotypes of recognised nephritogenic potential were isolated from five patients.


Deep brain stimulation of the subthalamic nucleus in severe Parkinson's disease: relationships between dual-contact topographic setting and 1-year worsening of speech and gait.
Subthalamic nucleus (STN) deep brain stimulation (DBS) alleviates severe motor fluctuations and dyskinesia in Parkinson's disease, but may result in speech and gait disorders. Among the suspected or demonstrated causes of these adverse effects, we focused on the topography of contact balance (CB; individual, right and left relative dual positions), a scantly studied topic, analyzing the relationships between symmetric or non-symmetric settings, and the worsening of these signs. An observational monocentric study was conducted on a series of 92 patients after ethical approval. CB was specified by longitudinal and transversal positions and relation to the STN (CB sub-aspects) and totalized at the patient level (patient CB). CB was deemed symmetric when the two contacts were at the same locations relative to the STN. CB was deemed asymmetric when at least one sub-aspect differed in the patient CB. Baseline and 1-year characteristics were routinely collected: (i) general, namely, Unified Parkinson's Disease Rating Scores (UPDRS), II, III motor and IV, daily levodopa equivalent doses, and Parkinson's Disease Questionnaire of Quality of Life (PDQ39) scores; (ii) specific, namely scores for speech (II-5 and III-18) and axial signs (II-14, III-28, III-29, and III-30). Only significant correlations were considered (p < 0.05). Baseline characteristics were comparable (symmetric versus asymmetric). CB settings were related to deteriorations of speech and axial signs: communication PDQ39 and UPDRS speech and gait scores worsened exclusively with symmetric settings; the most influential CB sub-aspect was symmetric longitudinal position. Our findings suggest that avoiding symmetric CB settings, whether by electrode positioning or shaping of electric fields, could reduce worsening of speech and gait.


Dark Suite: a comprehensive toolbox for computer-aided drug design.
Dark Suite is a complete interactive software pipeline aiming to exploit the advantages of free software and modern programming. Besides two commands (installation and opening) on the command line, the handling and the program operation is done through a user's friendly interface. This platform has a central graphical interface which allows the user to choose in what computational tool to work. Each computational tool has an interface. Dark Suite combines the functions of other programmes to create a pipeline for high-quality secondary effects through a friendly user interface. It is made to run on GNU / Linux distributions and its interface was built using JAVA to seamlessly integrate scientific tools written in Perl, Java, R and Python.


An unusual case of accidental poisoning: fatal methadone inhalation.
In this report, the authors present a case of unusual, accidental methadone intoxication in a 40-year-old man, who had inhaled methadone powder. The drug dealer was a pharmacy technician; methadone had been stolen from a pharmacy and sold as cocaine. After having inhaled methadone powder, he suffered cardiopulmonary arrest. He was admitted to hospital where he died after 24 h of intensive care. The autopsy revealed congestion of internal organs and cerebral and pulmonary edema. Microscopically, the heart showed no changes. The toxicological analyses performed on blood and urine taken at the hospital revealed methadone, cannabinoids, and ethanol. The blood methadone concentration was 290 μg/L. The urine methadone concentration was 160 μg/L. Midazolam and lidocaine, which were administered to the patient at the hospital, were also detected in the blood. The cause of death was determined to be methadone intoxication. The literature has been reviewed and discussed. To date, and to our knowledge, only very few cases of accidental death resulting from methadone inhalation have been described up to the case presented herein.


Physiological properties and receptive fields of mechanosensory neurones in the head ganglion of the leech: comparison with homologous cells in segmental ganglia.
A study of the head ganglion of the leech was made to compare the properties of specific sensory cells in this ganglion with those of homologous neurones in the segmental ganglia. 1. In the head ganglion, cells were identified that had electrical properties, sensory modalities and adaptation properties similar to those of touch (T), pressure (P) and nociceptive (N) cells in the segmental ganglia. The cell bodies of these neurones were situated in characteristics positions that could be correlated with those in the segmental ganglia. Several lines of evidence suggested that they were primary sensory neurones. Fewer T, P and N neurones were identified in the head ganglion than would be expected from its six constituent segmental ganglia. 2. The receptive fields of identified T, P and N cells were situated on the external surface of the head and the interior of the mouth with considerable overlap. They were generally smaller in size than those situated on the main part of the body. The receptive fields were also displaced anteriorly so that some of them were situated in segments anterior to those of the innervating cells. 3. The morphology of the sensory cells in the head ganglion was studied by intracellular injection of horseradish perioxidase. The general branching characteristics of the cells and the structural appearance of their processes resembled those of homologous cells in the segmental ganglia. However, the routes taken to the periphery by some of the cells were not constant from head ganglion to head ganglion. This variability was confirmed by electrophysiological evidence, and differed from the constancy seen in segmental sensory cells.


A phase II trial of oral gimatecan for recurrent glioblastoma.
Gimatecan is a lipophilic oral camptothecin analogue with preclinical activity in glioma models. We conducted a multicenter phase II trial to evaluate the efficacy of gimatecan in adults with recurrent glioblastoma. Eligibility criteria included ≤1 prior treatment for recurrent disease, age ≥18, Eastern Cooperative Oncology Group performance status 0-1, and normal organ function. Patients taking enzyme-inducing anti-seizure medications were excluded. Gimatecan 1.22 mg/m(2) was given orally once daily for 5 consecutive days during each 28-day cycle. The primary endpoint was progression-free survival at 6 months. A Simon 2-stage optimal design was used in which 19 patients were evaluated in the 1st stage, with an additional 36 patients accrued if >4 patients in stage 1 achieved PFS at 6 months. 29 patients were enrolled in the study, with median age of 58 years (range, 25-77 years); 58.6 % female. All patients received prior surgery, radiation therapy, and at least one chemotherapy regimen. The daily dose was reduced to 1.0 mg/m(2) after four of the first 10 patients experienced grade 4 hematologic toxicity. Treatment-related grade 3/4 toxicities included thrombocytopenia (17.2 %), leukopenia (17.2 %) and neutropenia (10.3 %). None of the 19 patients treated at 1.0 mg/m(2)/day experienced grade 4 hematologic toxicity. One patient had a partial radiographic response by modified Macdonald criteria. Only 3 patients (12 %) were progression-free at 6 months. Median time to progression was 12.0 weeks (7.0, 17.0).Treatment with gimatecan 1.0 mg/m(2)/day for 5 days, repeated every 28-days showed minimal efficacy.


Angiographic Findings of an Isolated Meandering Pulmonary Vein: A Case Report.
A meandering pulmonary vein (MPV) is a rare pulmonary vascular anomaly characterized by an abnormal course of the pulmonary vein draining into the left atrium. We report the case of a 55-year-old female who was diagnosed with a right MPV on pulmonary angiography. Enhanced chest CT revealed a vascular structure with an abnormal course that drained into the right superior pulmonary vein in the right upper lobe, which resembled a pulmonary arteriovenous malformation (AVM). Pulmonary angiography performed to discriminate between an MPV and pulmonary AVM showed no feeding artery, normal parenchymal staining, and drainage to the left atrium via the culprit vessel. Thus, the patient was diagnosed with MPV. When an MPV is difficult to differentiate from other vascular anomalies on enhanced chest CT, pulmonary angiography can be helpful. By recognizing the angiographic findings of an MPV, unnecessary treatment can be prevented.


Effects of Smoking Cannabis on Visual Function and Driving Performance. A Driving-Simulator Based Study.
Cannabis is the most widely used illegal drug in the world. Limited information about the effects of cannabis on visual function is available, and more detail about the possible impact of visual effects on car driving is required. This study investigated the effects of smoking cannabis on vision and driving performance, and whether these effects are correlated. Twenty drivers and occasional users were included (mean (SE) age, 23.3 (1.0) years; five women). Vision and simulated driving performance were evaluated in a baseline session and after smoking cannabis. Under the influence of cannabis, certain visual functions such as visual acuity (p < 0.001), contrast sensitivity (p = 0.004) and stereoacuity (far, p < 0.001; near, p = 0.013) worsened. In addition, there was an overall deterioration of driving performance, with the task of keeping the vehicle in the lane proving more difficult (p < 0.05). A correlation analysis showed significant associations between driving performance and visual function. Thus, the strongest correlations were found between the distance driven onto the shoulder and stereoacuity, for near (ρ = 0.504; p = 0.001) and far distances (ρ = 0.408; p = 0.011). This study provides the first evidence to show that the visual effects of cannabis could impact driving performance, compromising driving safety. The results indicate that information and awareness campaigns are essential for reducing the incidence of driving under the influence of cannabis.


[Home medical care service and regional medical plan].
A home medical service should be promoted as an individual medical plan which proactively copes with health problems in daily living. It is categorized into 3 phases: Health for a patient without long term disabilities, QOL for a patient with long term disabilities and Peaceful Death for a terminal patient.


Magnetic resonance-based techniques for the study and management of multiple sclerosis.
Conventional magnetic resonance imaging (cMRI) is widely used for diagnosing multiple sclerosis (MS) and for monitoring its activity and evolution. However, the correlation between cMRI and clinical findings of MS is limited, possibly due to the low pathological specificity of the abnormalities seen on cMRI scans and to the inability of cMRI to quantify the extent of the damage of the normal-appearing tissues. Magnetization transfer and diffusion-weighted MRI can quantify the extent and pathological severity of structural changes occurring within and outside cMRI-visible MS lesions. Proton MR spectroscopy can add information on the biochemical nature of such changes. Finally, functional MRI can provide new insights into the role of cortical adaptive changes in limiting the clinical consequences of MS structural damage. The application of quantitative MR-based techniques is changing dramatically our understanding of how MS causes irreversible disability and there is increasing perception that these methodologies should be more extensively employed in clinical trials to derive innovative information.


Effectiveness of probiotics on the duration of illness in healthy children and adults who develop common acute respiratory infectious conditions: a systematic review and meta-analysis.
Recent systematic reviews have reported a positive, although modest, effect of probiotics in terms of preventing common cold symptoms. In this systematic review, the effect of probiotics, specifically Lactobacillus and Bifidobacterium strains, on the duration of acute respiratory infections in otherwise healthy children and adults was evaluated. To identify relevant trials, eight databases, including MEDLINE, Embase, the Cochrane Database of Systematic Reviews (CDSR), the Cochrane Central Register of Controlled Trials (CENTRAL), the Database of Abstracts of Reviews of Effects (DARE), Health Technology Assessment (HTA), Science Citation Index (SCI) and OAISTER, were searched from inception to 20 July 2012. Details regarding unpublished studies/databases were also obtained from probiotic manufacturers. Study selection, data extraction and quality assessment were carried out by two reviewers. Risk of bias was assessed using criteria adapted from those published by the Centre for Reviews and Dissemination. In this review, twenty randomised controlled trials (RCT) were included, of which twelve were considered to have a low risk of bias. Meta-analysis revealed significantly fewer numbers of days of illness per person (standardised mean difference (SMD) - 0·31 (95% CI - 0·41, - 0·11), I²= 3%), shorter illness episodes by almost a day (weighted mean difference - 0·77 (95% CI - 1·50, - 0·04), I²= 80%) (without an increase in the number of illness episodes), and fewer numbers of days absent from day care/school/work (SMD - 0·17 (95% CI - 0·31, - 0·03), I²= 67%) in participants who received a probiotic intervention than in those who had taken a placebo. Reasons for heterogeneity between the studies were explored in subgroup analysis, but could not be explained, suggesting that the effect sizes found may differ between the population groups. This systematic review provides evidence from a number of good-quality RCT that probiotics reduce the duration of illness in otherwise healthy children and adults.


Chronic widespread pain as a medico-legal issue.
A systematic approach to the determination of disability in persons with chronic widespread pain is based on a careful history, examination for tender points, and special investigations to establish a firm diagnosis. Further assessment requires determination of subjective and objective measures of impairment. The causes of fibromyalgia are multifactorial and understanding them requires an appreciation of the role of physical or psychological trauma, and associated medical and psychosocial factors. Despite the presence of disability risk factors, most patients with fibromyalgia maintain a good range of normal daily activities and continue working.


Novel Therapeutics in Psoriatic Arthritis. What Is in the Pipeline?
To highlight the recently approved therapeutic agents in psoriatic arthritis (PsA), drugs in the pipeline, as well as to discuss efficacy with regard to different clinical domains of PsA. More than 15 years ago, tumor necrosis factor inhibitors (TNFi) were the first biologic disease modifying anti-rheumatic drugs (bDMARDs) that were approved for the treatment of PsA. Since then, multiple new therapeutic agents inhibiting other targets have emerged including biologics targeting interleukin (IL) 12/23, and IL 17 and oral agents targeting phosphodiesterase 4 (PDE4) and Janus kinases (JAKs). Many new agents with various modes of action including selective inhibition of IL 23, therapies promoting activated T cell apoptosis, inhibition of tyrosine kinase 2 (TYK2), and more are under active assessment in ongoing clinical trials. Effective therapies for treating PsA have emerged over the last 15 years and newer agents continue to be discovered, allowing greater therapeutic options for controlling psoriatic disease activity and preventing joint damage and disability. Personalized therapy for patients with PsA is now a possibility.


The microbial flora of herring fillets after storage in carbon dioxide, nitrogen or air at 2 degrees C.
Herring fillets from the Baltic Sea were stored in glass vessels in air, nitrogen or carbon dioxide (CO2) at 2 degrees C and the microbial development was studied. The microbiological shelf-life of the herring (the time to reach 10(7) organisms/g) was prolonged by a factor of 3.5 in CO2 as compared to air. The corresponding factor in nitrogen was 1.5. The microflora of fresh and spoiled herring was classified. The initial microflora was dominated by coryneforms, Flavobacterium spp., Moraxella-like organisms and Pseudomonas spp. The spoilage-flora in air (after 9 d) was dominated by Pseudomonas spp. and Moraxella-like organisms, and in nitrogen (14 d) Enterobacteriaceae, Vibrionaceae and Lactobacillus spp. were dominant. Homofermentative Lactobacillus spp. were the only organisms isolated from fillets stored in CO2 (28 d). It was concluded that storing fresh fish in pure CO2 at low refrigeration temperatures is a method with industrial potential. The method (1) improves the microbiological stability and (2) reduces the microbiological health hazards of the fish.


[Eumycetic mycetoma due to Madurella mycetomatis. Report of six cases.].
We report six cases of mycetoma due to Madurella mycetomatis. Five men and a woman from 28 to 70 years of age, and a history of one to ten years. In four of them the foot was affected, in another the wrist, and one exceptional case with neck involvement after an iguana bite. The response to treatment was irregular and not satisfactory.


Expression and mutagenesis of human poly(ADP-ribose) polymerase as a ubiquitin fusion protein from Escherichia coli.
The cDNA of human poly(ADP-ribose) polymerase (pADPRP), encoding the entire protein, was subcloned into the Escherichia coli expression plasmid pYUb. In this expression system, the carboxyl terminus of ubiquitin is fused to the amino terminus of a target protein, in this case pADPRP, stabilizing the accumulation of the cloned gene product. Following induction of the transformed cells, the sonicated extract contained a unique protein immunoreactive with both pADPRP and ubiquitin antibodies and corresponding to the predicted mobility of the fusion protein in SDS-PAGE. Fusion of ubiquitin to pADPRP increased the yield of pADPRP approximately 10-fold compared to that of the unfused enzyme. The resulting recombinant fusion protein had catalytic properties which were nearly identical to those of native pADPRP obtained from mammalian tissues. These properties included specific activity, Km for NAD, response to DNA strand breaks, response to Mg2+, inhibition by 3-aminobenzamide, and activity in activity gel analysis. An initial analysis by deletion mutagenesis of pADPRP's functional domains revealed that deletions in the NAD binding domain eliminated all activity; however, partial polymerase activity resulted from deletion in the DNA binding or automodification domains. The activities were not enhanced by breaks in DNA. We further report a colony filter screening procedure designed to identify functional polymerase molecules which will facilitate structure/function studies of the polymerase.


Consortia modulation of the stress response: proteomic analysis of single strain versus mixed culture.
The high complexity of naturally occurring microbial communities is the major drawback limiting the study of these important biological systems. In this study, a comparison between pure cultures of Pseudomonas reinekei sp. strain MT1 and stable community cultures composed of MT1 plus the addition of Achromobacter xylosoxidans strain MT3 (in a steady-state proportion 9:1) was used as a model system to study bacterial interactions that take place under simultaneous chemical and oxidative stress. Both are members of a real community isolated from a polluted sediment by enrichment in 4-chlorosalicylate (4CS). The analysis of dynamic states was carried out at the proteome, metabolic profile and population dynamic level. Differential protein expression was evaluated under exposure to 4CS and high concentrations of toxic intermediates (4-chlorocatechol and protoanemonin), including proteins from several functional groups and particularly enzymes of aromatic degradation pathways and outer membrane proteins. Remarkably, 4CS addition generated a strong oxidative stress response in pure strain MT1 culture led by alkyl hydroperoxide reductase, while the community showed an enhanced central metabolism response, where A. xylosoxidans MT3 helped to prevent toxic intermediate accumulation. A significant change in the outer membrane composition of P. reinekei MT1 was observed during the chemical stress caused by 4CS and in the presence of A. xylosoxidans MT3, highlighting the expression of the major outer membrane protein OprF, tightly correlated to 4CC concentration profile and its potential detoxification role.


Response of glandular versus basal rat ventral prostatic epithelial cells to androgen withdrawal and replacement.
Light microscope autoradiography was utilized to investigate the separate proliferative activities of the basal and glandular cell types during androgen-mediated restoration of the ventral prostatic epithelium in castrated rats. Our objective was to determine whether the proliferative population of the prostatic epithelium is restricted to the basal cell subtype under these experimental conditions. In the intact animal, substantial numbers of basal as well as glandular cells were found to be proliferating. The basal cell labeling index (LI) was 3.75% +/- 0.3% or 1.85 +/- 0.4 x 10(5) labeled cells per prostate. For the glandular cells the comparable figures were 0.38% and 3.32 +/- 1.5 x 10(5). Seven days after surgical castration, both subtypes persisted. However, only the numbers of glandular cells were found to be significantly reduced (66% decrease). When androgens were replaced by daily subcutaneous injection (testosterone propionate, 2 mg/0.2 ml sesame oil), both cell types responded with significantly increased proliferation. The temporal pattern of response displayed by each cell type was very similar. A greater proportion of basal than glandular cells responded, ie, 28% vs 11% LI. However, in terms of approximate numbers of cells labeled during the peak response, the value for the glandular cells was larger, ie, 2.4 vs 1.1 x 10(6) labeled cells. These data clearly indicate that under normal conditions as well as during testosterone stimulation, basal cells are not the sole population of proliferating cells in the prostatic epithelium. Thus, basal cells, as a distinct morphologic population, are not homogeneously undifferentiated stem cells which give rise to differentiated and nonproliferating glandular epithelial cells. Aspects of these data can nevertheless be interpreted to support the idea that the prostatic epithelium is maintained by a single cell lineage system in which the stem cells comprise a subpopulation of the basal cells. Alternatively, the two populations may be separate and distinct as no data are presented which unequivocally links the two cell types in a precursor-progeny relationship.


On the insensitivity of single field planar dosimetry to IMRT inaccuracies.
To report on the sensitivity of single field planar measurements in identifying IMRT plans with poor calculational accuracy. Three IMRT plans for head and neck cancer were subjected to extensive quality assurance. The plans were recalculated on a cylindrical phantom and between eight and 18 low gradient points were measured in each plan with an ion chamber. Every point measured in these plans agreed to within 4% of the dose predicted by the planning system and the plans were judged acceptable for clinical use. Each plan was then reoptimized with aggressive dose constraints so that the new treatment fields were more highly modulated than the ones from the original plans. Very complex fields can be calculated less accurately and ion chamber measurements of these plans in the cylindrical phantom confirmed significant dosimetric errors--Several of the measured points in each plan differed from the calculated dose by more than 4%, with a maximum single deviation of 10.6%. These three plans were judged unacceptable for clinical use. All six plans (three acceptable, three unacceptable) were then analyzed with two means of individual field planar dosimetry: Portal imaging with an electronic portal imaging device (EPID) and an ion chamber array. Gamma analysis was performed on each set of planar measurements with 2%/2 mm distance to agreement (DTA) and 3%/3 mm DTA criteria to try to determine a gamma analysis threshold which would differentiate the flawed plans from the acceptable ones. With the EPID and 2%/2 mm DTA criteria, between 88.2% and 92.8% of pixels from the acceptable IMRT plans passed the gamma analysis, and between 87.5% and 91.9% passed for the unacceptable IMRT plans. With the ion chamber array and 2%/2 mm DTA criteria, between 92.4% and 94.9% of points in the acceptable plans passed the gamma analysis, while 86.8% to 98.3% of the points in the unacceptable plans passed the gamma analysis. The difference between acceptable and unacceptable plans was diminished further when gamma criteria were expanded to 3%/3 mm DTA. A fraction of pixels passing the gamma analysis was found to be a poor predictor of dosimetric accuracy with both planar dosimeters, as well as both sets of gamma criteria. Deconstruction of an IMRT plan for field-by-field QA requires complex analysis methods such as the gamma function. Distance to agreement, a component of the gamma function, has clinical relevance in a composite plan but when applied to individual, highly modulated fields, it can mask important dosimetric errors. While single field planar dosimetry may comprise one facet of an effective QA protocol, gamma analysis of single field measurements is insensitive to important dosimetric inaccuracies of the overall plan.


Burden and Patterns of Multimorbidity: Impact on Disablement in Older Adults.
The aim of this study was to assess the impact of the burden and patterns of multimorbidity on disability domains. In a cross-sectional study of 425 older adults from the Boston Rehabilitative Impairment Study of the Elderly, participants self-reported 13 chronic conditions and underwent assessment of body function (leg strength, velocity, and power, trunk extensor endurance, leg range of motion, foot sensation), activities (400-m walk test, Short Physical Performance Battery, Late Life Function and Disability Instrument function scores) and participation (Late Life Function and Disability Instrument participation scores). The association between multimorbidity patterns (identified by latent class analysis) and disablement measures, as well as multimorbidity burden (captured by a multimorbidity score) and disablement measures, was tested. Latent class analysis identified three classes-low multimorbidity, high multimorbidity, and predominantly musculoskeletal conditions. Class membership (multimorbidity pattern) was not associated with disablement measures, but multimorbidity score was associated with poor performance in all domains. A 1-point higher multimorbidity score was associated with lower scores in body functions (by 0.06 SD unit), activities (0.07-0.10 SD units), as well as participation (0.07-0.09 units). Multimorbidity counts may be an excellent tool for risk stratification and identification of persons in need of rehabilitation. Complete the self-assessment activity and evaluation online at http://www.physiatry.org/JournalCME CME OBJECTIVES: Upon completion of this article, the reader should be able to (1) describe and distinguish the effect of multimorbidity burden and multimorbidity patterns on three disability domains in older adults; (2) identify and discuss possible reasons why high multimorbidity burden may result in a restriction among social participation in older adults; and (3) detect disability risk among older patients during clinical assessment by using a simple count of common chronic conditions. Advanced ACCREDITATION: The Association of Academic Physiatrists is accredited by the Accreditation Council for Continuing Medical Education to provide continuing medical education for physicians.The Association of Academic Physiatrists designates this Journal-based CME activity for a maximum of 1.0 AMA PRA Category 1 Credit(s)™. Physicians should only claim credit commensurate with the extent of their participation in the activity.


Very Distal Femoral Periprosthetic Fractures: Replacement Versus Fixation: A Systematic Review.
To synthesize all-cause reoperations and complications data as well as secondary clinical and functional outcomes, after the management of very distal femur periprosthetic fractures (vDFPFs) in a geriatric patient population with either a distal femoral locking plate (DFLP) or distal femoral replacement (DFR). MEDLINE, Embase, and Web of Science were searched for English language articles from inception to March 16, 2020, in accordance to the PRISMA guidelines. Studies reporting the management of vDFPFs in adults older than 65 years with either a DFLP or DFR were included. To ensure this review solely focused on very distal femoral periprosthetic fractures, only fractures of the following classifications were included: (1) Lewis and Rorabeck type II or III, (2) Su and Associates' Classification of Supracondylar Fractures of the Distal Femur type III, (3) Backstein et al type F2, and/or (4) Kim et al type II or III. Three reviewers independently extracted data from the included studies. Study validity was assessed using the methodological index for nonrandomized studies (MINORS), a quality assessment tool for nonrandomized controlled studies in surgery. Twenty-five studies with 649 vDFPFs were included for analysis. There were 440 knees in the DFLP group (mean age range: 65.9-88.3 years) and 209 knees in the DFR group (mean age range: 71.0-84.8 years). Because of the literature's heterogeneity, the data were qualitatively synthesized. vDFPFs in the elderly treated with DFR underwent fewer reoperations relative to DFLP (0%-45% vs. 0%-77%, respectively). Time to weight-bearing was observably shorter in DFR studies relative to DFLP studies. Functional outcomes and postoperative range of motion indicated a trend for DFLP knees to outperform DFR knees. Future research should include prospective studies and cost-effectiveness evaluations to better understand the utility of DFR for these fractures. Therapeutic Level III. See Instructions for Authors for a complete description of levels of evidence.


New treatment possibilities in rheumatoid arthritis.
It is very difficult to predict future treatment modalities especially in diseases like rheumatoid arthritis (RA) with unknown etiology and pathogenesis. In the near future, traditional disease-modifying antirheumatic drugs (DMARD) alone, in combination with each other, or together with cyclosporine, FK506, Rapamycin, or Leflunomide, will probably be the main treatment for RA. Currently biological anti-TNFalpha agents like humanized MAb and recombinant TNF-receptor constructs are now launched in the market. This therapy alone, or in combination with methotrexate is very effective in RA patients. There are, however, concerns over increase in serious infections. Autologous stem cell transplantation will probably be used in certain patient with serious autoimmune diseases.


Isolation and characterization of a seed-specific isoform of microsomal omega-6 fatty acid desaturase gene (FAD2-1B) from soybean.
In plants, the endoplasmic reticulum (ER)-associated oleate desaturase (FAD2) is the key enzyme responsible for the production of linoleic acid in non-photosynthetic tissues. In this study, we report the characterization of a seed-specific isoform of microsomal omega-6 fatty acid desaturase gene (FAD2-1B) sharing high sequence similarity with FAD2-1 from soybean. Several potential promoter elements including seed-specific motifs are found in the 5'-flanking region of FAD2-1B gene. The ORF of FAD2-1B is 1161 bp long and encodes a protein of 387 amino acids. This deduced protein holds three histidine boxes and four putative membrane-spanning helices, and possesses a signal for endoplasmic reticulum retention at C-terminal. Yeast cells transformed with the plasmid construct containing soybean FAD2-1B accumulate an appreciable amount of linoleic acid (18:2), normally not present in wild-type yeast cells, indicating that the cloned gene encodes a functional FAD2 enzyme. Both semi-quantitative RT-PCR and in silico analysis show that FAD2-1B gene is specifically expressed in developing seeds of soybean.


Boron isotope evidence for oceanic carbon dioxide leakage during the last deglaciation.
Atmospheric CO2 fluctuations over glacial-interglacial cycles remain a major challenge to our understanding of the carbon cycle and the climate system. Leading hypotheses put forward to explain glacial-interglacial atmospheric CO2 variations invoke changes in deep-ocean carbon storage, probably modulated by processes in the Southern Ocean, where much of the deep ocean is ventilated. A central aspect of such models is that, during deglaciations, an isolated glacial deep-ocean carbon reservoir is reconnected with the atmosphere, driving the atmospheric CO2 rise observed in ice-core records. However, direct documentation of changes in surface ocean carbon content and the associated transfer of carbon to the atmosphere during deglaciations has been hindered by the lack of proxy reconstructions that unambiguously reflect the oceanic carbonate system. Radiocarbon activity tracks changes in ocean ventilation, but not in ocean carbon content, whereas proxies that record increased deglacial upwelling do not constrain the proportion of upwelled carbon that is degassed relative to that which is taken up by the biological pump. Here we apply the boron isotope pH proxy in planktic foraminifera to two sediment cores from the sub-Antarctic Atlantic and the eastern equatorial Pacific as a more direct tracer of oceanic CO2 outgassing. We show that surface waters at both locations, which partly derive from deep water upwelled in the Southern Ocean, became a significant source of carbon to the atmosphere during the last deglaciation, when the concentration of atmospheric CO2 was increasing. This oceanic CO2 outgassing supports the view that the ventilation of a deep-ocean carbon reservoir in the Southern Ocean had a key role in the deglacial CO2 rise, although our results allow for the possibility that processes operating in other regions may also have been important for the glacial-interglacial ocean-atmosphere exchange of carbon.


Eruptive purpuric papules on the arms; a case of chemotherapy-induced inflammation of actinic keratoses and review of the literature.
Chemotherapy-induced inflammation of actinic keratosis can present in patients with subclinical actinic keratoses that become erythematous and pruritic within weeks of initiating systemic chemotherapy. The reaction is limited to sun-exposed areas and, classically, histologic findings of parakeratosis and epidermal necrosis with keratinocyte nuclear pleomorphism are present. Exuberant reactions with extensive epidermal necrosis may lead to subepidermal vesiculation. We report a case of a 67-year-old man with a history of chronic hepatitis B virus infection and recently diagnosed squamous cell carcinoma of the lung who was noted to have progressive asymptomatic violaceous papules on the extensor forearms and distal upper arms while hospitalized for possible sepsis following initiation of chemotherapy. A dermatology consulatation was requested to rule out possible vasculitis. It is important to recognize chemotherapy-induced inflammation of actinic keratoses in predisposed patients; it may be managed successfully with topical corticosteroids and does not necessitate discontinuation of the offending chemotherapeutic agent.


Timing of cardiac magnetic resonance and diagnostic yield in patients with myocardial infarction with nonobstructive coronary arteries.
The present study sought to establish the diagnostic yield of cardiovascular magnetic resonance (CMR) in a large cohort of patients admitted with myocardial infarction (MI) with nonobstructive coronary artery disease (MINOCA) based on the timing of referral to CMR. Consecutive patients referred to CMR from January 2009 to February 2022 with a working diagnosis of MINOCA were retrospectively evaluated. Cine, T2-weighted, early, and late gadolinium-enhanced images were acquired and analyzed. The frequency of the underlying diagnosis and the association between timing of CMR and relative frequency of each diagnosis were assessed. We included 207 patients (median age 50 years, 60% men). Final diagnosis after CMR was achieved in 91% of the patients (myocarditis in 45%, MI in 20%, tako-tsubo cardiomyopathy in 19%, and other cardiomyopathies in 7%). The performance of CMR within 7 days of admission with MINOCA (median, 5 days; 117 patients) allowed a higher diagnostic yield compared with CMR performed later (median, 10 days; 88 patients) (96% vs 86%, P = .02). Although myocarditis was the most frequent diagnosis in both groups according to time to CMR, its frequency was higher among patients with a CMR performed within the first 7 days (53% vs 35%, P = .02). The frequency of other underlying diagnoses was not influenced by CMR timing. CMR led to an underlying diagnosis of MINOCA in 91% of patients and its diagnostic yield increased to 96% when CMR was performed within 7 days of admission. The most frequent diagnosis was myocarditis..


Ketamine monoanaesthesia for major surgery in epidermolysis bullosa. Case report.
A case report is presented of a patient with recessive dystrophic epidermolysis bullosa, who required anaesthesia for a major plastic surgery operation. The special anaesthetic problems are discussed, and ketamine monoanaesthesia is advocated when muscle relaxation is unnecessary. Glycopyrrolate was found to be an excellent antisialogenic agent.


A possible case of position effect of DNA replication in Drosophila melanogaster.
The behavior of T(3;4)10 provides evidence that the centromere of 4 is not in the doublet 101D3,4, and that most of what has been called the left arm of 4 should be called basal 4R. Basal 4R usually does not replicate as much as the tip of 4R when 4 is in its normal position in the chromocenter. Translocated to the tip of 3R, however, basal 4R attains a width equal to that of the rest of the arm-a possible position effect on DNA replication.


Surface order and stability of langmuir-blodgett films.
Angstrom-resolution atomic force microscope images of Langmuir-Blodgett monolayers and multilayers of cadmium arachidate in air and under water show a dramatic change from a disordered arrangement to a crystalline lattice by the addition or removal of a single layer of molecules. The disordered surface is less stable than the ordered one to mechanical stresses such as atomic force microscopy tip forces or at the air-water contact line during contact angle measurements. The difference in the degree of order in the alkyl chains is attributed to the strong attractive interaction between headgroups in the presence of the divalent cation.


Boron in plants: a biochemical role.
Boron, as borate, appears to have a role in partitioning metabolism between the glycolytic and pentose-shunt pathways. This effect results from the association of borate with 6-phosphogluconic acid, forming a virtual substrate that inhibits the action of 6-phosphogluconate dehydrogenase. In the absence of borate, the inhibition of the enzyme is released, and excess phenolic acids are formed. These acids also associate strongly with borate and thus develop an autocatalytic system for production of excess phenolic acids which cause necrosis of tissue and eventual death of the plant.


Research progress of minimally invasive surgery for gastric cancer.
Since the first laparoscopic radical surgery for early gastric cancer 30 years ago, there has been a gradual shift from "open" to "minimally invasive" surgery for gastric cancer. This transition is due to advancements in refined anatomy, enlarged field of view, faster recovery, and comparable oncological outcomes. Several high-quality clinical studies have demonstrated the safety and effectiveness of laparoscopy in the treatment of both early and locally advanced gastric cancer. The role of perioperative chemotherapy in managing locally advanced gastric cancer has been widely recognized, and there have been continuous breakthroughs in the exploration of targeted therapy and immunotherapy for perioperative treatment. Additionally, the application of indocyanine green near-infrared imaging technology, 3D laparoscopic technology, and robotic surgery systems has further improved the accuracy and minimally invasive nature of gastric cancer surgeries. Looking ahead, the field of minimally invasive surgery for gastric cancer is expected to become more standardized, resulting in a significant enhancement in the quality of life for gastric cancer patients.


Physical inactivity at leisure and work: a 12-month study of cardiac patients.
Physical inactivity has been identified as a distinct health risk. However, little is known about how this can vary at leisure and work in cardiac patients. The aim of this study was to examine the prevalence and predictors of inactivity during leisure and work in the 12 months following a cardiac event in Australian cardiac patients. A total of 346 patients consecutively admitted to hospital with acute coronary syndrome or to undergo coronary artery bypass graft surgery were interviewed in hospital, and 4 and 12 months later. Leisure and occupational physical activity was measured using the Stanford Brief Activity Survey. Sociodemographic, psychosocial, and clinical data were also collected. The prevalence of leisure-time physical inactivity declined over time, with 52% inactive preevent and 29% inactive at 12 months. Approximately 50% of participants were physically inactive in their work, regardless of whether this was measured before or after the cardiac event. Logistic regression revealed that the significant predictors of leisure-time physical inactivity at 12 months were non-home ownership (OR = 2.19; P = .007) and physical inactivity in leisure-time prior to the event (OR = 2.44; P = .001). The significant predictors of occupational physical inactivity at 12 months were white-collar occupation (OR = 3.10; P < .001) and physical inactivity at work prior to the event (OR = 12.99; P < .001). Preevent physical inactivity, socioeconomic, and clinical factors predicted both leisure and work inactivity after an acute cardiac event. Effective interventions could be designed and implemented to target those most at risk of being physically inactive at work or leisure.


Longitudinal imaging of Alzheimer pathology using [11C]PIB, [18F]FDDNP and [18F]FDG PET.
[(11)C]PIB and [(18)F]FDDNP are PET tracers for in vivo detection of the neuropathology underlying Alzheimer's disease (AD). [(18)F]FDG is a glucose analogue and its uptake reflects metabolic activity. The purpose of this study was to examine longitudinal changes in these tracers in patients with AD or mild cognitive impairment (MCI) and in healthy controls. Longitudinal, paired, dynamic [(11)C]PIB and [(18)F]FDDNP (90 min each) and static [(18)F]FDG (15 min) PET scans were obtained in 11 controls, 12 MCI patients and 8 AD patients. The mean interval between baseline and follow-up was 2.5 years (range 2.0-4.0 years). Parametric [(11)C]PIB and [(18)F]FDDNP images of binding potential (BP(ND)) and [(18)F]FDG standardized uptake value ratio (SUVr) images were generated. A significant increase in global cortical [(11)C]PIB BP(ND) was found in MCI patients, but no changes were observed in AD patients or controls. Subsequent regional analysis revealed that this increase in [(11)C]PIB BP(ND) in MCI patients was most prominent in the lateral temporal lobe (p < 0.05). For [(18)F]FDDNP, no changes in global BP(ND) were found. [(18)F]FDG uptake was reduced at follow-up in the AD group only, especially in frontal, parietal and lateral temporal lobes (all p < 0.01). Changes in global [(11)C]PIB binding (ρ = -0.42, p < 0.05) and posterior cingulate [(18)F]FDG uptake (ρ = 0.54, p < 0.01) were correlated with changes in Mini-Mental-State Examination score over time across groups, whilst changes in [(18)F]FDDNP binding (ρ = -0.18, p = 0.35) were not. [(11)C]PIB and [(18)F]FDG track molecular changes in different stages of AD. We found increased amyloid load in MCI patients and progressive metabolic impairment in AD patients. [(18)F]FDDNP seems to be less useful for examining disease progression.


Influence of Nd:YAG and diode laser irradiation on apical sealing when associated with AH plus and EndoREZ endodontic cements.
This study aims to evaluate the influence of Nd:YAG and diode laser irradiation on apical sealing when applied before root canal filling done with two different resin-based cements (AH Plus and EndoREZ). Lasers have been widely used in endodontics. The morphologic changes in dentin walls caused by Nd:YAG and diode laser irradiation could improve sealing ability of endodontic cements. Eighty canine teeth were analyzed by apical leakage of methylene blue dye and by scanning electron microscope (SEM) images of the filling's adaptation to the apical third dentin walls. The laser irradiation parameters for the Nd:YAG laser were 100 mJ, 15 Hz, and 1.5 W, and for the diode laser was 2.5 W (continuous mode). The apical leakage results showed statistically significant differences at the 5% level of significance (Tukey's test, p < 0.05) among the different root canal wall treatments, independently of the root canal filling cement used. The cements also showed significant differences between them on the lased groups. The SEM analysis revealed better filling adaptation for AH Plus and the Nd:YAG laser group, confirming the microleakage results. We concluded that the combination of the Nd:YAG laser and AH Plus showed the best results.


Androgen Deprivation Therapy and the Risk of Anemia in Men with Prostate Cancer.
The use of androgen deprivation therapy in prostate cancer may be associated with an increased risk of anemia, but the evidence remains limited. This study aimed to determine if androgen deprivation is associated with increased risk of anemia in patients newly diagnosed with prostate cancer. This was a population-based cohort study using the United Kingdom Clinical Practice Research Datalink linked to the Hospital Episode Statistics repository. The cohort consisted of 10,364 men newly diagnosed with nonmetastatic prostate cancer between 1 April 1998 and 30 September 2015. We used time-dependent Cox proportional hazards models to estimate adjusted hazard ratios (HRs) and 95% confidence intervals (CIs) for anemia (hemoglobin <130 g/L) associated with current and past use of androgen deprivation therapy, compared with nonuse. There were 3,651 incident anemia events during 31,574 person-years of follow-up (rate: 11.6/100 person-years). Current androgen deprivation therapy use was associated with a nearly three-fold increased hazard of anemia, compared with nonuse (23.5 vs. 5.9 per 100 person-years, respectively; HR: 2.90, 95% CI: 2.67, 3.16). The HR was elevated in the first 6 months of use (HR: 2.20, 95% CI: 1.95, 2.48) and continued to be elevated with longer durations of use. Past androgen deprivation therapy use was associated with a lower estimate (HR: 1.27, 95% CI: 1.12, 1.43), which returned closer to the null ≥25 months after treatment discontinuation (HR: 0.95, 95% CI: 0.79, 1.15). The use of androgen deprivation therapy is associated with increased risk of anemia, which reverses upon treatment discontinuation.


An Exploration of Discrepancies and Concordances Between Hospital Disaster Directors and General Health Care Providers in Gyeonggi Province, South Korea: Quantitative Analysis of a Multicenter Cross-Sectional Survey Study.
The purpose of this study was to investigate differences in the perception of disaster issues between disaster directors and general health care providers in Gyeonggi Province, South Korea. The Gyeonggi provincial committee distributed a survey to acute care facility personnel. Survey topics included awareness of general disaster issues, hospital preparedness, and training priorities. The questionnaire comprised multiple choices and items scored on a 10-point Likert scale. We analyzed the discrepancies and characteristics of the responses. Completed surveys were returned from 43 (67%) of 64 directors and 145 (55.6%) of 261 health care providers. In the field of general awareness, the topic of how to triage in disaster response showed the greatest discrepancies. In the domain of hospital level disaster preparedness, individual opinions varied most within the topics of incident command, manual preparation. The responses to "accept additional patients in disaster situation" showed the biggest differences (> 21 versus 6~10). In this study, there were disaster topics with discrepancies and concordances in perception between disaster directors and general health care providers. The analysis would present baseline information for the development of better training programs for region-specific core competencies, knowledge, and skills required for the effective response.


Evidence for mitochondrial localization of a novel human sialidase (NEU4).
Based on the human cDNA sequence predicted to represent the NEU4 sialidase gene in public databases, a cDNA covering the entire coding sequence was isolated from human brain and expressed in mammalian cells. The cDNA encodes two isoforms: one possessing an N-terminal 12-amino-acid sequence that is predicted to be a mitochondrial targeting sequence, and the other lacking these amino acids. Expression of the isoforms is tissue specific, as assessed by reverse transcription-PCR. Brain, muscle and kidney contained both isoforms; liver showed the highest expression, and the short form was predominant in this organ. In transiently transfected COS-1 cells, enzyme activity was markedly increased with gangliosides as well as with glycoproteins and oligosaccharides as substrates compared with the control levels. This differs from findings with other human sialidases. Although the isoforms were not distinguishable with regard to substrate specificity, they exhibited differential subcellular localizations. Immunofluorescence microscopy and biochemical fractionation demonstrated that an exogenously expressed haemagglutinin-tagged long form of NEU4 was concentrated in mitochondria in several human culture cell types, whereas the short form was present in intracellular membranes, indicating that the sequence comprising the N-terminal 12 amino acid residues acts as a targeting signal for mitochondria. Co-localization of the long form to mitochondria was further supported by efficient targeting of the N-terminal region fused to enhanced green fluorescent protein, and by the targeting failure of a mutant with an amino acid substitution in this region. NEU4 is possibly involved in regulation of apoptosis by modulation of ganglioside G(D3), which accumulates in mitochondria during apoptosis and is the best substrate for the sialidase.


[Introduction of the electronic health card in Germany: influence of interest positions and sector membership on the establishment of an implementation network].
This article raises the question as to which mode of governance has been established for the implementation of the electronic health card (eHC) in Germany and which coordination problems have emerged. It will be shown that the amalgamation of two different policy domains - namely health-care and innovation policy - leads to a novel governance configuration in which information exchange (hence governance) is affected by sector membership and interest similarity as opposed to anticipated financial burdens or influence reputation in the newly established policy domain. The study is based on a standardised survey of 29 organisations that are involved in the implementation of the eHC. The analysis is carried out using quantitative methods such as social network analysis.


Advances in the diagnosis of lung cancer: contribution of molecular biology to bronchoscopic diagnosis.
Considerable advances in genomics, transcriptomics and proteomics have in the recent years transformed our understanding of the molecular mechanisms involved in the pathogenesis of lung cancer and are in the process of revolutionizing our approach to its diagnosis and treatment. Although these techniques have traditionally been described in the context of large volume biopsies from surgically resected tumors, significant technical advances allow their application to smaller samples obtained bronchoscopically, often the only samples available in advanced lung cancer. Recent data support the use of advanced molecular techniques as an adjunct to conventional histologic examinations of bronchoscopy specimens in the following situations: early diagnosis and screening of nonsmall cell lung cancer by fluorescent in-situ hybridization techniques in particular; accurate histologic diagnosis via novel immunohistochemistry markers; and prognosis and prediction of response to targeted and conventional chemotherapeutic agents. Molecular biology techniques are increasingly applied to smaller biopsy specimens obtained via bronchoscopy, allowing for their use in advanced, unresectable nonsmall cell lung cancer. Although these techniques have not yet entered the clinical practice arena, they will likely become an unavoidable complement to conventional morphologic examinations and allow for individualized diagnostic and therapeutic approach to lung-cancer patients.


Organ distribution of 99m-Tc-and 51-Cr-labelled thymocytes.
We have employed 99mTc as a radioisotopic label to study the organ distribution of murine thymocytes. The compartmentalization of 99mTc-labeled cells that had not been reduced by treatment with stannous chloride was similar to that of 51Cr-labeled cells and was characterized initially by 48-50% uptake of the injected radioactivity by the lungs. Increased hepatic and splenic and decreased pulmonary localization were noted at 1 hr and these shifts were more pronounced by 4 hr. Technetium-99m-labeled cells reduced by stannous chloride had significantly different patterns of hepatic, pulmonary, and splenic localization and at 4 hr the lungs still retained 24% of the injected radioactivity compared with only 3% in the spleen. Size distribution studies revealed that unlabeled as well as unreduced and reduced 99mTc-labeled thymocytes were almost identical to one another so that differences in compartmentalization could not be attributed to this factor. Since reduction with stannous chloride did not alter the distribution of 51Cr-labeled cells, this suggested some type of complex interaction between stannous ions and the labeling species of 99mTc. The in vivo localization of intravenously administered Na99mTcO4 and Na2CrO4 was markedly different from the corresponding radiolabeled cells thereby indicating that the distribution patterns that we observed truly represented cell-associated radioactivity. Although it may not necessarily be the proper reference point, the similarity in organ distribution of 99mTc- and 51Cr-labeled cells should allow direct comparison of previously reported data employing this radionuclide and that obtained in future studies with 99mTc.


A human TRIM5alpha B30.2/SPRY domain mutant gains the ability to restrict and prematurely uncoat B-tropic murine leukemia virus.
Human TRIM5alpha restricts N-tropic murine leukemia virus (N-MLV) but not B-tropic MLV (B-MLV) infection. Here we study B30.2/SPRY domain mutants of human TRIM5alpha that acquire the ability to inhibit B-MLV infection prior to reverse transcription without losing the ability to restrict N-MLV infection. Remarkably, these mutants gain the ability to decrease the amount of particulate B-MLV capsids in the cytosol of infected cells. In addition, these mutants gain the ability to restrict SIV(mac) and HIV-2 infection. B-MLV and SIV(mac) infections were blocked by the mutant TRIM5alpha proteins prior to reverse transcription. Thus, the range of retroviruses restricted by human TRIM5alpha can be increased by changes in the B30.2/SPRY domain, which also result in the ability to cause premature uncoating of the restricted retroviral capsid.


Self-monitoring has potential for home exercise programmes in patients with haemophilia.
Haemophiliacs who have had to keep a physically inactive lifestyle due to bleeding during childhood are likely to have little motivation for exercise. The purpose of this study is to clarify the effectiveness of the self-monitoring of home exercise for haemophiliacs. A randomized controlled trial was conducted with intervention over 8 weeks at four hospitals in Japan. Subjects included 32 male outpatients aged 26-64 years without an inhibitor who were randomly allocated to a self-monitoring group and a control group. Individual exercise guidance with physical activity for improvement of their knee functions was given to both groups. The self-monitoring materials included an activity monitor and a feedback system so that the self-monitoring group could send feedback via the Internet and cellular phone. The self-monitoring was performed by checking exercise adherence and physical activity levels, bleeding history and injection of a coagulation factor. Both groups showed significant improvements in exercise adherence (P < 0.001) and physical function such as the strength of knee extension (P < 0.001), range of knee extension (P < 0.001), range of ankle dorsiflexion (P < 0.01), a modified Functional Reach (P < 0.05) and 10 metre gait time (P < 0.01). In particular, improvements in exercise adherence (P < 0.05), self-efficacy (P < 0.05), and strength of knee extension (P < 0.05) were significant in the self-monitoring group compared with those in the control group. No increase in bleeding frequency and pain scale was noted. The self-monitoring of home exercise for haemophilic patients is useful for the improvement of exercise adherence, self-efficacy and knee extension strength.


Assessment of MR stereotactic imaging and image co-registration accuracy for 3 different MR scanners by 3 different methods/phantoms: phantom and patient study.
OBJECTIVEThe aim of this study was to compare 3 different methods to assess the geometrical distortion of two 1.5-T and one 3-T magnetic resonance (MR) scanners and to evaluate co-registration accuracy. The overall uncertainty of each particular method was also evaluated.METHODSThree different MR phantoms were used: 2 commercial CIRS skull phantoms and PTGR known target phantom and 1 custom cylindrical Perspex phantom made in-house. All phantoms were fixed in the Leksell stereotactic frame and examined by a Siemens Somatom CT unit, two 1.5-T Siemens (Avanto and Symphony) MRI systems, and one 3-T Siemens (Skyra) MRI system. The images were evaluated using Leksell GammaPlan software, and geometrical deviation of the selected points from the reference values were determined. The deviations were further investigated for both definitions including fiducial-based and co-registration-based in the case of the CIRS phantom images. The same co-registration accuracy assessment was also performed for a clinical case. Patient stereotactic imaging was done on 3-T Skyra, 1.5-T Avanto, and CT scanners.RESULTSThe accuracy of the CT scanner was determined as 0.10, 0.30, and 0.30 mm for X, Y, and Z coordinates, respectively. The total estimated uncertainty in distortion measurement in one coordinate was determined to be 0.32 mm and 0.14 mm, respectively, for methods using and not using CT as reference imaging. Slightly more significant distortions were observed when using the 3-T than either 1.5-T MR units. However, all scanners were comparable within the estimated measurement error. Observed deviation/distortion for individual X, Y, and Z stereotactic coordinates was typically within 0.50 mm for all 3 scanners and all 3 measurement methods employed. The total radial deviation/distortion was typically within 1.00 mm. Maximum total radial distortion was observed when the CIRS phantom was used; 1.08 ± 0.49 mm, 1.15 ± 0.48 mm, and 1.35 ± 0.49 mm for Symphony, Avanto, and Skyra, respectively. The co-registration process improved image stereotactic definition in a clinical case in which fiducial-based stereotactic definition was not accurate; this was demonstrated for 3-T stereotactic imaging in this study. The best results were shown for 3-T MR image co-registration with CT images improving image stereotactic definition by about 0.50 mm. The results obtained with patient data provided a similar trend of improvement in stereotactic definition by co-registration.CONCLUSIONSAll 3 methods/phantoms used were evaluated as satisfactory for the image distortion measurement. The method using the PTGR phantom had the lowest uncertainty as no reference CT imaging was needed. Image co-registration can improve stereotactic image definition when fiducial-based definition is not accurate.


Precise Lipidomics Decipher Circulating Ceramide and Sphingomyelin Cycle Associated with the Progression of Rheumatoid Arthritis.
Rheumatoid arthritis (RA) is a long-term autoimmune condition that causes joint and surrounding tissue inflammation. Lipid mediators are involved in inflammation and deterioration of the joints. Despite attempts to discover effective drug targets to intervene with lipid metabolism in the disease, progress has been limited. In this study, precise lipidomic technology was employed to quantify a broad range of serum ceramides and sphingomyelin (SM) in a large cohort, revealing an association between the accumulation of circulating ceramides and disturbed ceramide/SM cycles during the progression of RA. In our investigation, we discovered that eight ceramides exhibited a positive correlation with the activity of RA, thereby enhancing the accuracy of RA diagnosis, particularly in patients with serum antibody-negative RA. Furthermore, the enzyme SM phosphodiesterase 3 (SMPD3) was found to disrupt the circulating SM cycle and accelerate the progression of RA. The activity of SMPD3 can be inhibited by methotrexate, resulting in decreased metabolic conversion of SM to ceramide. These findings suggest that targeting the SM cycle may provide a new therapeutic option for RA.