[PCR-based detection of heteroplasmic deleted mitochondrial DNA in Kearns-Sayre syndrome].
To describe the clinical data and the results of molecular analyses of the mitochondrial DNA in a patient with Kearns-Sayre Syndrome. Molecular analyses of mitochondrial DNA from the patient included PCR amplification of a region where the common Kearns- Sayre deletion is located and Genotype-Phenotype correlations are discussed. The affected patient showed ptosis, progressive external ophthalmoplegia, pigmentary changes in the peripheral retina and right bundle block. Molecular analysis disclosed a approximately 5 kb deletion in the mitochondrial DNA and some wild type mtDNA indicating heteroplasmy. Molecular analysis of mitochondrial DNA confirmed the clinical diagnosis of Kearns-Sayre syndrome. PCR provides a rapid method to identify the common 4997 bp deletion in Kearns-Sayre syndrome. In such cases, PCR diagnosis could avoid invasive methods such as muscle biopsy or spinal tap.


Phage display epitope mapping of human neutrophil flavocytochrome b558. Identification of two juxtaposed extracellular domains.
Despite extensive experimental and clinical evidence demonstrating the critical role of flavocytochrome b558 (Cyt b) in the NADPH-dependent oxidase, there is a paucity of direct structural data defining its topology in the phagocyte membrane. Unlike other Cyt b-specific monoclonal antibodies, 7D5 binds exclusively to an extracellular domain, and identification of its epitope should provide novel insight into the membrane topology of Cyt b. To that end, we examined biochemical features of 7D5-Cyt b binding and used the J404 phage display nonapeptide library to identify the bound epitope. 7D5 precipitated only heterodimeric gp91-p22phox and not individual or denatured Cyt b subunits from detergent extracts of human neutrophils and promyelocytic leukemia cells (gp91-PLB). Moreover, 7D5 precipitated precursor gp65-p22phox complexes from detergent extracts of the biosynthetically active gp91-PLB cells, demonstrating that complex carbohydrates were not required for epitope recognition. Epitope mimetics selected from the J404 phage display library by 7D5 demonstrated that (226)RIVRG(230) and (160)IKNP(163) regions of gp91phox were both bound by 7D5. These studies reveal specific information about Cyt b membrane topology and structure, namely that gp91phox residues (226)RIVRG(230) and (160)IKNP(163) are closely juxtaposed on extracytoplasmic domains and that predicted helices containing residues Gly(165)-Ile(190) and Ser(200)-Glu(225) are adjacent to each other in the membrane.


TP53, p14ARF, p16INK4a and H-ras gene molecular analysis in intestinal-type adenocarcinoma of the nasal cavity and paranasal sinuses.
Intestinal-type adenocarcinoma (ITAC) of the nasal cavity and paranasal sinuses is an uncommon tumor associated with occupational exposure to dusts of different origin. Few investigations addressed molecular alterations in ITAC mainly focused on TP53, K-ras and H-ras gene mutations. The occurrence of TP53, p14(ARF) and p16(INK4a) deregulation and H-ras mutations was investigated in 21 consecutive and untreated ITACs cases, 17 with known professional exposure. No H-ras mutations were found. In patients with known exposure, cumulative evidence of TP53 or p14(ARF) alterations accounted for 88% and the evidence of p16(INK4a) alterations for 65%, respectively. TP53 mutations were present in 44% of the ITACs, consisted of G:C-->A:T transitions in 86%, and involved the CpG dinucleotides in 50% of the cases. LOH at the locus 17p13 and an uncommon high rate of p53 stabilization were detected in 58% and 59% of the cases, respectively. p14(ARF)and p16(INK4a) promoter methylation accounted for 80% and 67% respectively, and LOH at the locus 9p21 occurred in 45% of the cases. Interestingly, all dust-exposed tumors with p16(INK4a) alterations shared TP53 or p14(ARF) deregulation. The present results show a close association of this occupational tumor with TP53, p14(ARF) and p16(INK4a) gene deregulation. Given the important role that these genes play in cell growth control and apoptosis, the knowledge of ITAC genetic profile may be helpful in selecting more tailored treatments.


Modeling the corneal birefringence of the eye toward the development of a polarimetric glucose sensor.
Optical polarimetry for monitoring glucose concentration in the aqueous humor of the eye as a potential noninvasive means of assessing blood glucose has promise, but the realization of such an approach has been limited by noise from time-varying corneal birefringence due to motion artifact. Modeling the corneal birefringence of the eye is critically important toward understanding the overall effect of this noise source compared to other changes in the signal, and can aid in design of the polarimetric system. To this end, an eye model is introduced in this work that includes spatially varying birefringence properties of the cornea. The degree of birefringence and the fast axis orientation is calculated as a function of beam position on the anterior chamber. It is shown that the minimum change in polarization vector orientation occurs for beam position near the midpoint between the corneal apex and limbus. In addition, the relative wavelength independence of motion artifact is shown in the same region. The direct consequence of these findings are that a multiwavelength polarimetric system can potentially be utilized to eliminate the effect of time-varying corneal birefringence, and that eye coupling is optimal at the midpoint between the apex and limbus.


Paralemmin-1, a modulator of filopodia induction is required for spine maturation.
Dendritic filopodia are thought to participate in neuronal contact formation and development of dendritic spines; however, molecules that regulate filopodia extension and their maturation to spines remain largely unknown. Here we identify paralemmin-1 as a regulator of filopodia induction and spine maturation. Paralemmin-1 localizes to dendritic membranes, and its ability to induce filopodia and recruit synaptic elements to contact sites requires protein acylation. Effects of paralemmin-1 on synapse maturation are modulated by alternative splicing that regulates spine formation and recruitment of AMPA-type glutamate receptors. Paralemmin-1 enrichment at the plasma membrane is subject to rapid changes in neuronal excitability, and this process controls neuronal activity-driven effects on protrusion expansion. Knockdown of paralemmin-1 in developing neurons reduces the number of filopodia and spines formed and diminishes the effects of Shank1b on the transformation of existing filopodia into spines. Our study identifies a key role for paralemmin-1 in spine maturation through modulation of filopodia induction.


The effect of Benson Relaxation Technique on rheumatoid arthritis patients: extended report.
Rheumatoid arthritis (RA) is a chronic disease and has the highest rate of prevalence among rheumatic inflammatory diseases. The purpose of this study was to determine the effect of Benson Relaxation Technique (BRT) combined with medication on disease activity in patients with RA. Following ethical approval, 50 consecutive matched patients were selected and allocated into two groups, either an experimental or a control group. Patients in the experimental group received BRT combined with medication and patients in the control group were given only medication. Clinical symptoms, laboratory findings, anxiety, depression and feeling of well-being were measured before and after intervention to evaluate the effect of BRT. There was a significant difference between the two groups in anxiety, depression and feeling of well-being. Changes in clinical symptoms and laboratory findings were not large enough to be statistically significant between the two groups, but they indicated decline in disease progress. The results demonstrate that BRT can be an effective technique in reducing disease process in patients suffering from RA. However, in order to support the findings of this trial, studies with large sample size and > 8-week intervention are recommended.


[Comparison of data between intratracheal instillation and inhalation studies for estimation of harmful effects of manufactured nanomaterials].
We reviewed the difference in data between the inhalation and intratracheal instillation studies of inhaled materials and how to diminish the difference in data between the two studies in order to examine the usefulness of intratracheal instillation study for the estimation of the harmful effects of manufactured nanomaterials. The differences in the data of the intratracheal instillation study from those of the inhalation study, which is the gold standard, are as follows: 1) artificial effect of the bolus, 2) nonuniformity of lung distribution of materials, 3) no threshold of clearance, 4) low deposition level of materials in the brain and neonates, and 5) the effect of dispersant on the lung. We consider that the approaches to diminishing the difference in data between the two studies are as follows: 1) maintain the dispersion of nanomaterials, 2) avoid the overdosing of nanomaterials, and 3) maintain a low concentration of the dispersant in the intratracheal instillation study.


Isogenic lysogens of diverse shiga toxin 2-encoding bacteriophages produce markedly different amounts of shiga toxin.
We produced isogenic Escherichia coli K-12 lysogens of seven different Shiga toxin 2 (Stx2)-encoding bacteriophages derived from clinical Shiga toxin-producing E. coli (STEC) isolates of serotypes O157:H7, O145, O111, and O83 to assess the variability among these phages and determine if there were phage-related differences in toxin production. Phage genomic restriction fragment length polymorphisms (RFLP) and superinfection resistance studies revealed significant differences among these phages and allowed the seven phages to be placed into five distinct groups. Experiments revealed striking differences in spontaneous phage and toxin production that were correlated with the groupings derived from the RFLP and resistance studies. These results suggest that the genotype of the Stx2 prophage can influence the level of phage release and toxin expression by host strains and thus may be relevant to STEC pathogenesis.


Mode of death in Shapiro syndrome: a case report.
Shapiro syndrome is extremely rare and is characterized by the triad of spontaneous periodic hypothermia, hyperhidrosis and agenesis of the corpus callosum, resulting in neurological and psychological disorders. The exact mechanism of this syndrome is unknown and treatment consists of controlling the periodic attacks. This case report describes a case of Shapiro syndrome presenting with ventricular fibrillation (VF) who was treated with dual chamber implantable cardioverter defibrillator (ICD) therapy. A 45-year-old man, suffering from Shapiro syndrome with frequent hypothermic attacks, was admitted to the emergency department with an out of hospital cardiac arrest caused by VF due to hypothermia. To prevent cardiac death during future hypothermic attacks with VF, the patient was treated with a dual chamber ICD. Within 1 month after ICD implantation the patient had two events of ventricular tachycardia/VF during hypothermia, which were both successfully terminated by an ICD shock. One year after ICD implantation the patient suffered from an uncontrolled urinary tract infection and the patient passed away. Post-mortem interrogation of the ICD did not reveal further episodes of VF and showed a higher supraventricular heartrate in the last days before his death, probably due to a sinus tachycardia driven by the infection. It was concluded that the most likely cause of death was an uncontrolled sepsis. The current case showed that ICD therapy can be successful in treating VF episodes in patients with unexpected periods of hypothermia.


Bacterial membranes as predictors of antimicrobial potency.
A wide range of chemical structures having antimicrobial activity have been studied in an effort to treat the increasing emergence of bacteria that are resistant to traditional antibiotics. These agents have varying degrees of toxicity against different bacterial species. We demonstrate, using members of a novel class of antimicrobial agents, the oligomers of acyllysine, that one cause for the difference in species selectivity is the ability to induce the clustering of anionic lipids, resulting in their segregation into domains. This phenomenon occurs only in bacterial membranes composed of both anionic and zwitterionic lipids and not with bacteria whose membrane lipids are largely anionic. As a consequence it can be predicted which bacterial species will be most affected by antimicrobial agents that function principally by this mechanism. This finding allows for the design of new antibiotics with selective toxicity against different groups of bacteria.


TBM Hunter: Identify and Score Canonical, Extended, and Unconventional Tankyrase-Binding Motifs in Any Protein.
Tankyrases, a versatile protein group within the poly(ADP-ribose) polymerase family, are essential for post-translational poly(ADP-ribosyl)ation, influencing various cellular functions and contributing to diseases, particularly cancer. Consequently, tankyrases have become important targets for anti-cancer drug development. Emerging approaches in drug discovery aim to disrupt interactions between tankyrases and their binding partners, which hinge on tankyrase-binding motifs (TBMs) within partner proteins and ankyrin repeat cluster domains within tankyrases. Our study addresses the challenge of identifying and ranking TBMs. We have conducted a comprehensive review of the existing literature, classifying TBMs into three distinct groups, each with its own scoring system. To facilitate this process, we introduce TBM Hunter-an accessible, web-based tool. This user-friendly platform provides a cost-free and efficient means to screen and assess potential TBMs within any given protein. TBM Hunter can handle individual proteins or lists of proteins simultaneously. Notably, our results demonstrate that TBM Hunter not only identifies known TBMs but also uncovers novel ones. In summary, our study offers an all-encompassing perspective on TBMs and presents an easy-to-use, precise, and free tool for identifying and evaluating potential TBMs in any protein, thereby enhancing research and drug development efforts focused on tankyrases.


Imatinib mesylate in children and adolescents with cancer.
Imatinib is an inhibitor of the BCR-ABL fusion gene product that characterizes chronic myeloid leukemia (CML), and of the related tyrosine kinases c-KIT and platelet-derived growth factor (PDGF) receptor. The drug is now included as front-line therapy for CML and Philadelphia chromosome-positive acute lymphoblastic leukemia in children and adolescents, though valid concerns about serious late sequelae remain unresolved and are important issues for further study. European and North American consortia have conducted phase I and II clinical trials of imatinib in children and adolescents with brain and other solid tumors that have provided little evidence of efficacy.


Education in orthopaedic trauma.
Education of the orthopaedic surgeon in trauma care presents a unique challenge because of the need to acquire knowledge about the overall resuscitation of the patient as well as having a sound working knowledge of the treatment of injuries to other organ systems and how that treatment interplays with orthopaedic care in such complex disorders such as adult respiratory distress syndrome. The orthopaedic trauma educator requires special dedication and skill because much teaching occurs at night and on holidays and weekends in the context of emergency surgery where immediate decisions are required and procedures must be executed expeditiously and with minimal morbidity. An ideal educational program in trauma provides ready availability of the attending with close supervision of a team of residents lead by an experienced chief or fellow to whom appropriate graduated responsibility for decision making and execution of surgery is provided. A formal didactic course with a complete multidisciplinary curriculum is essential. The managed healthcare movement has the potential to disrupt the educational process and clinical research in orthopaedic trauma by reducing referrals to major trauma centers; by making high quality trauma treatment, particularly in the indigent population economically impractical; and by interfering with the patient care and educational and research process by early transfer of patients to other providers removed from the trauma center environment. Orthopaedic trauma educators must work with their institutions, national organizations, government, and the healthcare industry to develop methods to preserve and advance the education and research in trauma care which has served our population so well and is so important for the health and productivity of society in the future.


Translation inhibition reveals interaction of 2'-deoxy and 2'-O-methyl molecular beacons with mRNA targets in living cells.
Understanding the interaction between oligonucleotide probes and RNA targets in living cells is important for biological and clinical studies of gene expression in vivo. Here, we demonstrate that starvation of cells and translation inhibition by blocking the mTOR or PI-3 kinase pathway could significantly reduce the fluorescence signal from 2'-deoxy molecular beacons (MBs) targeting K-ras and GAPDH mRNAs in living cells. However, the intensity and localization of fluorescence signal from MBs targeting nontranslated 28S rRNA remained the same in normal and translation-inhibited cells. We also found that, in targeting K-ras and GAPDH mRNAs, the signal level from MBs with 2'-O-methyl backbone did not change when translation was repressed. Taken together, our findings suggest that MBs with DNA backbone hybridize preferentially with mRNAs in their translational state in living cells, whereas those with 2'-O-methyl chemistry tend to hybridize to mRNA targets in both translational and nontranslational states. This work may thus provide a significant insight into probe design for detection of RNA molecules in living cells and RNA biology.


Prediction of cervical infiltration in Stage II endometrial cancer by different preoperative evaluation techniques (D&C, US, CT, MRI).
Our clinical practice for FIGO Stage II endometrial cancer consists of Wertheim's radical hysterectomy as first choice of treatment. The evaluation of patients is based on D&C. The accuracy of this preoperative staging method is examined here. Twenty-nine patients with endometrial cancer with suspected cervical involvement (FIGO Stage II) based on endocervical curettage underwent Wertheim's radical hysterectomy between January 1, 1989 and December 31, 2001 at the Gynaecological Department of the National Institute of Cancer, Budapest, Hungary. In all cases surgico-pathological staging was performed to examine the accuracy of preoperative D&C and to find out whether radical surgery was necessary in all patients and how the preoperative evaluation of patients should be improved. Out of 29 patients who underwent Wertheim's hysterectomy the pathological examination found primary cervical cancer in two patients. These two patients were eliminated from further evaluation. Out of the remaining 27 patients only eight (29.6%) had cervical involvement of endometrial cancer documented by a pathologic review on the hysterectomy specimen. Extrauterine disease was documented in one of the patients with cervical infiltration (1/8) and in one in the cervix-negative group (1/19). Ovarian spread was found in the first case and ovarian infiltration with penetration of the tumour into the parametric tissue in the second case. According to the FIGO classification 18 (66.6%) patients had less extensive disease and two (7.4%) had more extensive disease. Only 26% of the patients (7/27) had surgical findings consistent with the Stage II disease. We can conclude that "overtreatment" seems to have occurred in 19 patients, whose cervical infiltration by endometrial cancer could not be proved by pathological staging. It can also be assessed that understaging took place in two cases, which can be explained by two reasons; we did not make use of preoperative imaging techniques since US was applied in six patients, CT in 16 and the most accurate, MRI, on three patients only. The other possible reason, which can point out the bad efficacy of the imaging techniques as well, could be that a major part of the patients received preoperative AL treatment, which could also have influenced the cervical progression. This is possible, but has not been proved. The difference in the number of cervical infiltrations in the group of patients who received preoperative radiotherapy and in the group where they did not, is not significant (p = 0.9742), and infiltration of the endometrium was present in all cases. In the future, proper selection of imaging modalities can improve the staging of gynaecological disorders and preclude unnecessary procedures. In endometrial cancer cases US, especially with the use of TVUS, is often considered to be the primary imaging approach. However, in patients where ultrasound is suboptimal, where there is a large tumour present or the result of imaging studies will directly influence the choice of therapy and guide therapy planning then the higher accuracy of MRI warrants its use. CT is of use in the later stages of disease; differentiation between Stage I and II is difficult and CT is limited in the assessment of myometrial invasion.


Shear-induced phase separation in solutions of wormlike micelles.
Polymer solutions in the vicinity of the theta-point are known to undergo shear-induced turbidity or phase separation. The present study shows that a similar phenomenon also occurs for certain wormlike micellar solutions. Wormlike micelles are the self-assembled counterparts of polymers and are characterized by their ability to reversibly break and recombine. In the system of interest, the micelles are formed by the cationic surfactant erucyl bis(hydroxyethyl)methylammonium chloride (EHAC), in conjunction with a salt such as sodium chloride (NaCl) or sodium salicylate (NaSal). Micellar samples that become turbid under shear show evidence of critical concentration fluctuations and may contain predominantly branched micelles. The shear-induced turbidity in these samples correlates with the appearance of flow-dichroism in rheooptic experiments and with an increase in low-q scattering in small-angle light scattering under flow (flow-SALS) experiments. The characteristic "butterfly" pattern, with enhanced scattering in the flow direction and a dark streak perpendicular to the flow direction, is typically observed in flow-SALS. The results suggest that the turbidity is due to a shear-induced growth of concentration fluctuations, which in turn manifests as large anisotropic domains, typically oriented along the vorticity axis.


Phosphatidylinositol 4,5-bisphosphate mediates Ca2+-induced platelet alpha-granule secretion: evidence for type II phosphatidylinositol 5-phosphate 4-kinase function.
To understand the molecular basis of granule release from platelets, we examined the role of phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P(2)) in alpha-granule secretion. Streptolysin O-permeabilized platelets synthesized PtdIns(4,5)P(2) when incubated in the presence of ATP. Incubation of streptolysin O-permeabilized platelets with phosphatidylinositol-specific phospholipase C reduced PtdIns(4,5)P(2) levels and resulted in a dose- and time-dependent inhibition of Ca(2+)-induced alpha-granule secretion. Exogenously added PtdIns(4,5)P(2) inhibited alpha-granule secretion, with 80% inhibition at 50 microm PtdIns(4,5)P(2). Nanomolar concentrations of wortmannin, 33.3 microm LY294002, and antibodies directed against PtdIns 3-kinase did not inhibit Ca(2+)-induced alpha-granule secretion, suggesting that PtdIns 3-kinase is not involved in alpha-granule secretion. However, micromolar concentrations of wortmannin inhibited both PtdIns(4,5)P(2) synthesis and alpha-granule secretion by approximately 50%. Antibodies directed against type II phosphatidylinositol-phosphate kinase (phosphatidylinositol 5-phosphate 4-kinase) also inhibited both PtdIns(4,5)P(2) synthesis and Ca(2+)-induced alpha-granule secretion by approximately 50%. These antibodies inhibited alpha-granule secretion only when added prior to ATP exposure and not when added following ATP exposure, prior to Ca(2+)-mediated triggering. The inhibitory effects of micromolar wortmannin and anti-type II phosphatidylinositol-phosphate kinase antibodies were additive. These results show that PtdIns(4,5)P(2) mediates platelet alpha-granule secretion and that PtdIns(4,5)P(2) synthesis required for Ca(2+)-induced alpha-granule secretion involves the type II phosphatidylinositol 5-phosphate 4-kinase-dependent pathway.


Amphiphilic and magnetic behavior of Fe3O4 nanocrystals.
Fe(3)O(4) (magnetite) nanocrystals with an average size of 13 nm were prepared using a specially synthesized polymeric stabilizer (poly-4-vinylpyridine-co-acrylamidoethylamine-co-polyethyleneglycol-co-polyethyleneglycolic acid) that bears reactive groups for further functionalization. The polymer shell made the nanocrystals amphiphilic, i.e. they are soluble in pentanol, water and dichloromethane, and they distribute over the three solvents in a three phase system. The Fe(3)O(4) nanocrystals exhibited the well-known and typical magnetic properties.


Repair of O(6)-alkylguanine by alkyltransferases.
The predominant pathway for the repair of O(6)-methylguanine in DNA is via the activity of an alkyltransferase protein that transfers the methyl group to a cysteine acceptor site on the protein itself. This review article describes recent studies on this alkyltransferase. The protein repairs not only methyl groups but also 2-chloroethyl-, benzyl- and pyridyloxobutyl-adducts. It acts on double-stranded DNA by flipping the O(6)-guanine adduct out of the DNA helix and into a binding pocket. The free base, O(6)-benzylguanine, is able to bind in this pocket and react with the cysteine, rendering it an effective inactivator of mammalian alkyltransferases. The alkylated form of the protein is rapidly degraded by the ubiquitin/proteasomal system. Some tumor cells do not express alkyltransferase despite having an intact gene. Methylation of key sites in CpG-rich islands in the promoter region are involved in this silencing and a change in the nuclear localization of an enhancer binding protein may also contribute. The alkyltransferase promoter contains Sp1, GRE and AP-1 sites and is slightly inducible by glucocorticoids and protein kinase C activators. There is a complex relationship between p53 and alkyltransferase expression with p53 mediating a rise in alkyltransferase in response to ionizing radiation but having no clear effect on basal levels. DNA adducts at the O(6)-position of guanine are a major factor in the carcinogenic, mutagenic, apoptopic and clastogenic actions of methylating agents and chloroethylating agents. Studies with transgenic mice in which alkyltransferase levels are increased or decreased confirm the importance of this repair pathway in protecting against carcinogenesis. Alkyltransferase activity in tumors protects them from therapeutic agents such as temozolomide and BCNU. This resistance is abolished by O(6)-benzylguanine and this drug is currently in clinical trials to enhance cancer chemotherapy by these agents. Studies are in progress to reduce the toxicity of such therapy towards the bone marrow by gene therapy to express alkyltransferases with mutations imparting resistance to O(6)-benzylguanine at high levels in marrow stem cells. Several polymorphisms in the human alkyltransferase gene have been identified but the significance of these in terms of alkyltransferase action is currently unknown.


Endovascular recanalization of the subacute to chronically occluded basilar artery: initial experience and technical considerations.
Occlusion of the basilar artery (BA) has a poor prognosis. We evaluated technical considerations and complications associated with reopening subacute to chronically occluded BAs. Duration of BA occlusion before revascularization, symptoms and medical management before treatment, and postprocedural antiplatelet regimen and anticoagulation protocols of 9 patients were analyzed. All patients underwent endovascular low-volume balloon angioplasty followed by Wingspan stenting. The median time between onset of symptoms and treatment was 5 days (range, 2 days to 3.5 years). The median time between documentation of BA occlusion by cerebral angiography or computed tomography angiography and treatment was 3 days (range, 1 day to 8 months). Recanalization was successful in 8 of the 9 patients. Immediately after the procedure, 4 patients were stable, 3 patients improved, and 2 patients were worse. Four patients had periprocedural complications. Four of the 9 patients died, 2 from periprocedural complications. The mean clinical duration of follow-up was 11 months. At latest follow-up, the modified Rankin Scale scores for the 5 surviving patients were 0, 0, 2, 2, and 3, respectively. During the follow-up period, 4 patients improved, 1 patient remained stable, and 1 patient died. The mean angiographic follow-up was 8.6 months. Two patients developed significant in-stent stenosis during this period. With current endovascular techniques, recanalization of chronically occluded BAs is feasible. The procedure carries substantial risks and should be reserved for patients with medically refractory symptoms. Careful postprocedural medical management and radiographic follow-up are warranted to prevent in-stent restenosis.


Chemotherapy activates cancer-associated fibroblasts to maintain colorectal cancer-initiating cells by IL-17A.
Many solid cancers display cellular hierarchies with self-renewing, tumorigenic stemlike cells, or cancer-initiating cells (CICs) at the apex. Whereas CICs often exhibit relative resistance to conventional cancer therapies, they also receive critical maintenance cues from supportive stromal elements that also respond to cytotoxic therapies. To interrogate the interplay between chemotherapy and CICs, we investigated cellular heterogeneity in human colorectal cancers. Colorectal CICs were resistant to conventional chemotherapy in cell-autonomous assays, but CIC chemoresistance was also increased by cancer-associated fibroblasts (CAFs). Comparative analysis of matched colorectal cancer specimens from patients before and after cytotoxic treatment revealed a significant increase in CAFs. Chemotherapy-treated human CAFs promoted CIC self-renewal and in vivo tumor growth associated with increased secretion of specific cytokines and chemokines, including interleukin-17A (IL-17A). Exogenous IL-17A increased CIC self-renewal and invasion, and targeting IL-17A signaling impaired CIC growth. Notably, IL-17A was overexpressed by colorectal CAFs in response to chemotherapy with expression validated directly in patient-derived specimens without culture. These data suggest that chemotherapy induces remodeling of the tumor microenvironment to support the tumor cellular hierarchy through secreted factors. Incorporating simultaneous disruption of CIC mechanisms and interplay with the tumor microenvironment could optimize therapeutic targeting of cancer.


A series of four transcatheter aortic valve replacement in failed Perceval valves.
In recent years, sutureless valves (SV) and rapid deployment valves (RDVs) have become interesting aortic valve substitutes, especially in minimally invasive aortic valve surgery, as they reduce cardio-pulmonary bypass and cross-clamp times. There are two valve types available, the sutureless Perceval and the rapid deployment Intuity valve prosthesis. When these valves fail, besides surgical re-replacement, the valve-in-valve concept has been reported in a small series of case reports. Our own experience includes four cases of failed Perceval valves, in which a balloon-expandable transcatheter valve was implanted in three patients, and a self-expanding transcatheter valve was implanted in a fourth patient. Here, we present these four cases with a focus on the specific valve design of the Perceval valve, as well as on important technical aspects. All cases were performed successfully with clinical improvement. Transcatheter aortic valve replacement (TAVR) as a valve-in-valve concept seems to be a valuable option in selected patients with failed sutureless or RDVs.


Dissipative Organization of DNA Oligomers for Transient Catalytic Function.
The development of synthetic non-equilibrium systems opens doors for man-made life-like materials. Yet, creating distinct transient functions from artificial fuel-driven structures remains a challenge. Building on our ATP-driven dynamic covalent DNA assembly in an enzymatic reaction network of concurrent ATP-powered ligation and restriction, we introduce ATP-fueled transient organization of functional subunits for various functions. The programmability of the ligation/restriction site allows to precisely organize multiple sticky-end-encoded oligo segments into double-stranded (ds) DNA complexes. We demonstrate principles of ATP-driven organization into sequence-defined oligomers by sensing barcode-embedded targets with different defects. Furthermore, ATP-fueled DNAzymes for substrate cleavage are achieved by transiently ligating two DNAzyme subunits into a dsDNA complex, rendering ATP-fueled transient catalytic function.


Recent status scores for version 6 of the Addiction Severity Index (ASI-6).
To describe the derivation of recent status scores (RSSs) for version 6 of the Addiction Severity Index (ASI-6). 118 ASI-6 recent status items were subjected to nonparametric item response theory (NIRT) analyses followed by confirmatory factor analysis (CFA). Generalizability and concurrent validity of the derived scores were determined. A total of 607 recent admissions to variety of substance abuse treatment programs constituted the derivation sample; a subset (n = 252) comprised the validity sample. The ASI-6 interview and a validity battery of primarily self-report questionnaires that included at least one measure corresponding to each of the seven ASI domains were administered. Nine summary scales describing recent status that achieved or approached both high scalability and reliability were derived; one scale for each of six areas (medical, employment/finances, alcohol, drug, legal, psychiatric) and three scales for the family/social area. Intercorrelations among the RSSs also supported the multi-dimensionality of the ASI-6. Concurrent validity analyses yielded strong evidence supporting the validity of six of the RSSs (medical, alcohol, drug, employment, family/social problems, psychiatric). Evidence was weaker for the legal, family/social support and child problems RSSs. Generalizability analyses of the scales to males versus females and whites versus blacks supported the comparability of the findings, with slight exceptions. The psychometric analyses to derive Addiction Severity Index version 6 recent status scores support the multi-dimensionality of the Addiction Severity Index version 6 (i.e. the relative independence of different life functioning areas), consistent with research on earlier editions of the instrument. In general, the Addiction Severity Index version 6 scales demonstrate acceptable scalability, reliability and concurrent validity. While questions remain about the generalizability of some scales to population subgroups, the overall findings coupled with updated and more extensive content in the Addiction Severity Index version 6 support its use in clinical practice and research.


Gender differences in mental health expectancies in early- and midlife in six European countries.
Health expectancies, taking into account both quality and quantity of life, have generally been based on disability and physical functioning. To compare mental health expectancies at age 25 and 55 based on common mental disorders both across countries and between males and females. Mental health expectancies were calculated by combining mortality data from population life tables and the age-specific prevalence of selected common mental disorders obtained from the European Study of the Epidemiology of Mental Disorders (ESEMeD). For the male population aged 25 (all countries combined) life expectancy was 52 years and life expectancy spent with a common mental disorder was 1.8 years (95% CI 0.7-2.9), 3.4% of overall life expectancy. In comparison, for the female population life expectancy at age 25 was higher (57.9 years) as was life expectancy spent with a common mental disorder (5.1 years, 95% CI 3.6-6.6) and as a proportion of overall life expectancy, 8.8%. By age 55 life expectancy spent with a common mental disorder had reduced to 0.7 years (males) and 2.3 years (females). Age and gender differences underpin our understanding of years spent with common mental disorders in adulthood. Greater age does not mean living relatively more years with common mental disorder. However, the female population spends more years with common mental disorders and a greater proportion of their longer life expectancy with them (and with each studied separate mental disorder).


[Ultrastructure of the proliferation of astrocytes and microglia].
Glial cell proliferation was studied during axonal reaction of hypoglossal nerve, and around stab wound in the brain cortex of the rat. The cytoplasm and chromosomes of astroglial mitoses were pale. Lipid droplets, few sparse dense bodies with heterogenous structure were present in the cytoplasm. The mitotic astrocytes had irregular outlines. The ultrastructure of "light microglial" cells was described; it was found that these cells divided and gave rise to microglial cells. The cytoplasm and the chromosomes of microglial mitoses were dense; the cytoplasm contained always groups of dense bodies and lipfuscine granules. The outlines of mitotic microglial cells were more regular.


Leveraging nanoscale plasmonic modes to achieve reproducible enhancement of light.
The strongly enhanced and localized optical fields that occur within the gaps between metallic nanostructures can be leveraged for a wide range of functionality in nanophotonic and optical metamaterial applications. Here, we introduce a means of precise control over these nanoscale gaps through the application of a molecular spacer layer that is self-assembled onto a gold film, upon which gold nanoparticles (NPs) are deposited electrostatically. Simulations using a three-dimensional finite element model and measurements from single NPs confirm that the gaps formed by this process, between the NP and the gold film, are highly reproducible transducers of surface-enhanced resonant Raman scattering. With a spacer layer of roughly 1.6 nm, all NPs exhibit a strong Raman signal that decays rapidly as the spacer layer is increased.


Airborne contaminant exposure control in a partitioned work environment by exhaust ventilation systems.
The evaluation of contaminant removal effectiveness can play a key role in a working environment quality investigation and in remediation efforts. In this study a computer simulation technique was developed for assessing the efficiency of contaminant removal within a partitioned working environment. To determine the effect of the physical parameters of each partitioned space on contaminant removal effectiveness, a relative contaminant concentration unit was used to show the simulation results. In addition, tracer gas techniques were adopted to validate the accuracy of the prediction model. A comparison of measured CO2 concentrations in a controlled environment chamber with the results of a simulation model is presented. This study investigated the partition configuration of work environment and environment parameters including openings operation, constant concentration, and exhaust air volume mode. Results indicated that variations of the above three parameters can produce great differences in overall ventilation performance. Through this type of study, the interaction of airflow and contaminant concentration between partitioned spaces can be understood in advance and adequate knowledge can be provided to maintain a high-quality and healthy environment for workers.


Understanding the particokinetics of engineered nanomaterials for safe and effective therapeutic applications.
Increasing numbers of engineered nanomaterials (ENMs) are being developed for therapeutic and diagnostic applications. However, the tunable and varied physicochemical properties of ENMs pose a new challenge for understanding their biological behavior, trafficking, and biodistribution. Herein the concept of "particokinetics" is introduced to address the dynamic biological behavior of ENMs at the molecular level (including gravitational sedimentation, dispersion, aggregation, and interaction with biomolecules in suspending media), cellular level (including cellular uptake, transport, biotransformation, and elimination), and whole-organism level (including absorption, distribution, metabolism, and excretion in vivo). Several mathematical modeling methods are introduced which guide a quantitative description of their biological behavior at different levels. Examples are also provided to delineate the impact of the physicochemical properties of ENMs on their particokinetics. A comprehensive understanding of the in vivo and in vitro particokinetics of ENMs will facilitate the design of tailor-made functional ENMs that act as highly effective and controllable drug-delivery systems with minimal side-effects.


High external phosphate (Pi) increases sodium ion uptake and reduces salt tolerance of 'Pi-tolerant' soybean.
Previous studies on the interaction between environmental inorganic phosphate (Pi) and salinity stress using soybean cultivars sensitive to high external Pi had two limitations: (1) the phenotype was dominated by overaccumulation of phosphorus (P); and (2) no detailed analysis was performed for sodium ion uptake. In this study, we focused on the effects of high external Pi on the sodium ion uptake in 'Pi-tolerant' soybean cultivars. The P accumulation in Pi-tolerant soybean Union was much lower [9.0 mg g(-)(1) dry weight (DW); contrasting to 38-76 mg g(-)(1) DW in the 'Pi-sensitive' soybean cultivars]. At in planta level, high level of external Pi significantly (P < 0.001) increased net sodium ion uptake and aggravated salinity stress symptoms. The effects of high external Pi diminished when de-rooted plants were used, suggesting that root is the primary organ interacting with Pi in the growth medium. Two-cell models, including soybean suspension cells and the tobacco Bright Yellow-2 cell line, were also employed to study the effects of high external Pi at the cellular level. Consistent to in planta results, high external Pi uplifted cellular sodium ion uptake and reduced cell viability under salinity stress. Gene expression analyses further showed that HPi (2 mM Pi supplements; excessive level of Pi) could reduce the fold of induction of GmSOS1 and GmCNGC under salinity stress, suggesting that they may be possible molecular targets involved in the interaction between high external Pi and Na(+) uptake.


Ab initio QM/MM study shows there is no general acid in the reaction catalyzed by 4-oxalocrotonate tautomerase.
The mechanism for the reaction catalyzed by the 4-oxalocrotonate tautomerase (4-OT) enzyme has been studied using a quantum mechanical/molecular mechanical (QM/MM) method developed in our laboratory. Total free energy barriers were obtained for the two steps involved in this reaction. In the first step, Pro-1 acts as a general base to abstract a proton from the third carbon of the substrate, 2-oxo-4-hexenedioate, creating a negative charge on the oxygen at C-2 of this substrate. In the second step, the same hydrogen abstracted by the N-terminal Pro-1 is shuttled back to the fifth carbon of the substrate to form the product, 2-oxo-3-hexenedioate. The calculated total free energy barriers are 14.54 and 16.45 kcal/mol for the first and second steps, respectively. Our calculations clearly show that there is no general acid in the reaction. Arg-39' ', which is hydrogen bonded to the carboxylate group of the substrate, and an ordered water, which moves closer to the site of the charge formed in the transition state and intermediate, play the main role in transition state/intermediate stabilization without acting as general acids in the reaction.


Catalytic reaction profile for NADH-dependent reduction of aromatic aldehydes by xylose reductase from Candida tenuis.
Kinetic substituent effects have been used to examine the catalytic reaction profile of xylose reductase from the yeast Candida tenuis, a representative aldo/keto reductase of primary carbohydrate metabolism. Michaelis-Menten parameters (k(cat) and K(m)) for NADH-dependent enzymic aldehyde reductions have been determined using a homologous series of benzaldehyde derivatives in which substituents in meta and para positions were employed to systematically perturb the properties of the reactive carbonyl group. Kinetic isotope effects (KIEs) on k(cat) and k(cat)/K(m) for enzymic reactions with meta-substituted benzaldehydes have been obtained by using NADH (2)H-labelled in the pro-R C4-H position, and equilibrium constants for the conversion of these aldehydes into the corresponding alcohols (K(eq)) have been measured in the presence of NAD(H) and enzyme. Aldehyde dissociation constants (K(d)) and the hydride transfer rate constant (k(7)) have been calculated from steady-state rate and KIE data. Quantitative structure-activity relationship analysis was used to factor the observed substituent dependence of k(cat)/K(m) into a major electronic effect and a productive positional effect of the para substituent. k(cat)/K(m) (after correction for substituent position) and K(eq) obeyed log-linear correlations over the substituent parameter, Hammett sigma, giving identical slope values (rho) of +1.4 to +1.7, whereas the same Hammett plot for logK(d) yielded rho=-1.5. This leads to the conclusion that electron-withdrawing substituents facilitate the reaction and increase binding to about the same extent. KIE values for k(cat) (1.8) and k(cat)/K(m) (2.7), and likewise k(7), showed no substituent dependence. Therefore, irrespective of the observed changes in reactivity over the substrate series studied no shift in the character of the rate-limiting transition state of hydride transfer occurred. The signs and magnitudes of rho values suggest this transition state to be product-like in terms of charge development at the reactive carbon. Structure-reactivity correlations reveal active-site homologies among NADPH-specific and dual NADPH/NADH-specific yeast xylose reductases and across two aldo/keto reductase families in spite of the phylogenetic separation of the host organisms producing xylose reductase (family 2B) and aldehyde reductase (family 1A).


[Introduction into PPPM: experience of the past and tomorrow's reality].
Health service today is on the verge of broad changes. The intensive practical application of the achievements of genomics, proteomics, metabolomics and bioinformatics has significantly deepened and continues to deepen our view of the pathological processes taking place at a level of biostructures. In the nearest future this progress will give medical practitioners the opportunity to focus on a subclinical stage of the disease, i.e. on the earliest stages of the pathological process. This will require the prediction of disease development risks, subclinical diagnostics with the precise staging of the pathological process, and, finally, the application as early as possible of targeted pharmacotherapeutic methods in order to prevent the manifestation of the disease or its progression into more severe stages. All these principles create the framework of a fundamentally new "3P" strategy in medicine: predictive, preventive and personalized medicine.


SNAIL expression correlates with the translocation of syndecan‑1 intracellular domain into the nucleus in prostate cancer cell lines.
Zinc finger protein SNAI1 (SNAIL) and zinc finger protein SNAI2 (SLUG) transcription factors promote epithelial‑mesenchymal transition, a process through which epithelial cells acquire a mesenchymal phenotype, increasing their migratory and invasive properties. In prostate cancer (PCa) progression, increased expression levels of SNAIL and SLUG have been described. In advanced PCa, a decrease in the cell surface proteoglycan syndecan‑1 (SDC‑1), which has a role in cell‑to‑extracellular matrix adhesion, has been observed. Notably, SDC‑1 nuclear location has been observed in mesenchymal cancers. The present study aimed to determine if SNAIL and SLUG may be associated with the nuclear location of SDC‑1 in PCa. To determine the location of SDC‑1, antibodies against its intracellular domain (ID) or extracellular domain (ED) were used in benign prostatic hyperplasia (BPH) and PCa samples with high Gleason scores. Only ID‑SDC‑1 was located in the cell nuclei in advanced PCa samples, but not in the BPH samples. ED‑SDC‑1 was located in the cell membrane and cytoplasm, exhibiting decreased levels in PCa in comparison with those in BPH. Furthermore, LNCaP and PC3 PCa cell lines with ectopic SNAIL expression exhibited nuclear ID‑SDC‑1. No change was observed in the ED‑SDC‑1 levels, and maintained its location in the cell membrane and cytoplasm. SLUG induced no change in ID‑SDC‑1 location. At the protein level, an association between SNAIL and nuclear ID‑SDC‑1 was observed. In conclusion, the results of the present study demonstrated that nuclear ID‑SDC‑1 localization was associated with SNAIL expression in PCa cell lines.


An ultra performance liquid chromatography-tandem mass spectrometry method for the therapeutic drug monitoring of isavuconazole and seven other antifungal compounds in plasma samples.
A new analytical method was developed for the routine Therapeutic Drug Monitoring of 8 antifungals compounds in 50μL of plasma: isavuconazole (ISZ), voriconazole (VRZ), posaconazole (PSZ), fluconazole (FCZ), caspofungin (CSF), flucytosine (5FC), itraconazole (ITZ) and its metabolite OH-itraconazole (OH-ITZ). After adding 50μL of the internal standard, which consisted in a mixture of the deuterated isotopes of the quantified compounds, the sample treatment consisted in a simple protein precipitation with 400μL of acetonitrile. Five microliters of the supernatant were directly injected into the chromatographic system. The chromatographic separation was performed with a Waters C18-BEH column and a mobile phase consisting in a mixture of water and acetonitrile, both containing 0.1% of formic acid. The total run time was 3min and the detection of the analytes was performed by electrospray ionization in a positive mode using selected reaction monitoring. Intra and inter-day precision and inaccuracy were <15% over the calibration ranges that were determined according to their clinical relevance: 0.20-20.0mg/L for ISZ, VRZ, PSZ, ITZ, and OH-ITZ; 0.50-50.0mg/L for FCZ and CSF; 2.00-200mg/L for 5FC. This simple and fast method was found suitable for routine therapeutic drug monitoring.


Evaluation of membrane bioreactor performance via residence time distribution: effects of membrane configuration and mixing.
Unlike conventional wastewater treatment systems that have a single effluent discharge point, membrane bioreactors (MBR) may have multiple extraction points resulting from the location of the membrane element in the reactor. This leads to multiple residence time distributions for an MBR system. One method to characterise the mixing is based on the concept of residence time distribution (RTD). A set of RTDs were generated using the conservative tracer, lithium chloride, for pilot plant MBRs with capacity up to 300 m3/day. Flat sheet and hollow fibre pilot plant MBR systems were operated in parallel on primary effluent collected at the Bedok Water Reclamation Plant in the republic of Singapore. Analysis of the RTD profiles indicated that membrane geometry did not impact on the kinetic conversion associated with nitrification because both MBRs were in well mixed conditions. However, the energy required to achieve perfect mixing with a hollow fibre module MBR, as defined by the velocity gradient, was lower than that with a flat sheet module MBR. The implication is that energy input associated with reactor mixing will depend on the configuration of the membrane. The difference in energy requirements between flat sheets and hollow fibres is such that careful consideration should be given to membrane selection in larger municipal installations.


The effect of physiological doses of thyroxine on the level of cyclic adenosine-3',5'-monophosphate in pituitary and anterior hypothalamus of male rats of different age.
The levels of cAMP in the pituitary and anterior hypothalamus were determined by competitive binding to protein in male rats aged 2--2.5, 6--8 and 14--18 months. The lowest level of cAMP both in the pituitary and anterior hypothalamus (1.05 +/- 0.17 and 0.85 +/- 0.23 pM) (per mg wet weight, respectively) was found in 6--8 month-old rats. Administration of T4 (2 micrograms/100 g body weight, for 8 days) was followed by a decrease in cAMP level in the pituitary of 14--18 month-old animals, which was not the case with younger rats. The obtained results agree with the earlier observation that the threshold to homeostatic suppression in the hypothalamus-pituitary-thyroid system of rats decreases with aging. It is suggested that cAMP system contributes to the mechanism of changes in hypothalamic threshold and its pharmacological correction.


Pain patient profile and the assessment of malingered pain.
This study investigated whether a pain sample and pain simulators could be distinguished on the Pain Patient Profile (P3). Forty patients with a pain condition completed the P3 under normal instructions, while 20 students responded under instructions to feign a pain disorder but to attempt to avoid detection. The simulators did not differ on the P3 Validity Scale compared with the pain group, but scored significantly higher than the pain group on the P3 clinical scales (Depression, Anxiety, Somatization). The simulators were more likely to obtain an abnormal score (T score > 55) on all of the clinical scales. The Depression scale had highest positive and negative predictive power and correctly classified 80% of the participants. The P3 may be a useful screening tool for assessing those feigning pain but requires further research.


Enzyme replacement therapy in late-onset Pompe's disease: a three-year follow-up.
Pompe's disease is an autosomal recessive myopathy. The characteristic lysosomal storage of glycogen is caused by acid alpha-glucosidase deficiency. Patients with late-onset Pompe's disease present with progressive muscle weakness also affecting pulmonary function. In search of a treatment, we investigated the feasibility of enzyme replacement therapy with recombinant human alpha-glucosidase from rabbit milk. Three patients (aged 11, 16, and 32 years) were enrolled in the study. They were all wheelchair-bound and two of them were ventilator dependent with a history of deteriorating pulmonary function. After 3 years of treatment with weekly infusions of alpha-glucosidase, the patients had stabilized pulmonary function and reported less fatigue. The youngest and least affected patient showed an impressive improvement of skeletal muscle strength and function. After 72 weeks of treatment, he could walk without support and finally abandoned his wheelchair. Our findings demonstrate that recombinant human alpha-glucosidase from rabbit milk has a therapeutic effect in late-onset Pompe's disease. There is good reason to continue the development of enzyme replacement therapy for Pompe's disease and to explore further the production of human therapeutic proteins in the milk of mammals.


Carcinogenic effects of acrylamide in Sencar and A/J mice.
Acrylamide structurally resembles vinyl carbamate, a proposed proximate carcinogenic form of ethyl carbamate. To test the hypothesis that acrylamide should possess carcinogenic properties, it was tested in the Salmonella-microsome assay for point mutation, as a skin tumor initiator in the Sencar mouse, and for its ability to induce lung adenomas in the A/J mouse. Acrylamide was found to be without activity as a mutagen in Salmonella strains TA 1535, TA 1537, TA 98, and TA 100 both in the presence and absence of rat liver microsomes using both the plate and liquid suspension assays. However, acrylamide was found to approximate ethyl carbamate in potency as a tumor initiator in the skin of the female Sencar mice. As with ethyl carbamate, acrylamide was more potent by systemic routes of administration relative to topical application. Acrylamide was also found to induce lung adenomas in male and female A/J mice using both the p.o. and i.p. routes of administration. Acrylamide was approximately one-seventh as potent as ethyl carbamate in the induction of lung adenomas. These data confirm the hypothesis that acrylamide possesses carcinogenic properties similar to ethyl carbamate.


Easily accessible C2-symmetric chiral bicyclo[3.3.0] dienes as ligands for rhodium-catalyzed asymmetric 1,4-addition.
The synthesis of a new type of C(2)-symmetric chiral diene ligands with a nonbridged bicyclic [3.3.0] backbone was successfully introduced. Using highly efficient lipase-catalyzed transesterification and Suzuki-coupling strategies, a broad family of enantiopure 3,6-disubstituted bicyclo[3.3.0] dienes with different electronic and steric properties could be easily prepared. The application of these new diene ligands in the Rh-catalyzed asymmetric 1,4-addition of arylboronic acids to alpha,beta-unsaturated carbonyl compounds have been examined, and excellent enantioselectivities (up to 98 % ee) as well as good yields are achieved under very mild reaction conditions at room temperature.


Characterization of carotenoids and vitamin E in R. rugosa and R. canina: Comparative analysis.
The hips of Rosa species have gained more attention in recent years due to their high contents of antioxidant compounds. This study was designed to compare rosehips of the two roses species Rosa rugosa and Rosa canina, including different products, on carotenoid contents, including phytoene and phytofluene, as well as vitamin E. The investigation allowed the identification and quantification of types of (Z)-isomers of lycopene and rubixanthin in both rosehips and focused also on isomerisation of both carotenoids. The carotenoid identification and quantification were done using HPLC-DAD and LC-MS/MS. The statistical analysis revealed significant differences (P<0.05) in carotenoid contents and (P<0.001) in vitamin E contents between different rosehips species. The HPLC analysis showed that carotenoid contents varied between rosehips species. The isomerisation of (all-E)-rubixanthin and (all-E)-lycopene using iodine-catalysed photoisomerisation showed that the (5'Z)-isomer gazaniaxanthin is the main (Z)-isomer of rubixanthin and the (13Z)-isomer is the main (Z)-isomer of lycopene.


KIT exon 17 mutations are predictive of inferior outcome in pediatric acute myeloid leukemia with RUNX1::RUNX1T1.
Pediatric core binding factor acute myeloid leukemia (CBF-AML), although considered a favorable risk subtype, exhibits variable outcomes primarily driven by additional genetic abnormalities, such as KIT mutations. In this study, we examined the prognostic impact of KIT mutations in 130 pediatric patients with CBF-AML, treated uniformly at a single center over 4 years (2017-2021). KIT mutations were detected via next-generation sequencing using a myeloid panel comprising 52 genes for most patients. Our findings revealed that KIT mutations were present in 31% of CBF-AML cases. Exon 17 KIT mutation was most commonly (72%) seen with notable occurrences at the D816 and N822 residue in 48% and 39% of cases, respectively. The 3-year cumulative incidence of relapse (CIR) and overall survival (OS) for patients with exon 17 KIT mutation were 36% and 40%, respectively, and was significantly worse in comparison to other site KIT mutations (3-year CIR: 11%; OS: 64%) and without KIT mutation (3-year CIR: 13%; OS:71%). Notably, the prognostic impact of KIT mutations was prominent in patients with RUNX1::RUNX1T1, but not in those with CBFB::MYH11 fusion. Additionally, a high KIT variant-allele frequency (VAF) (>33%) predicted for a higher disease relapse; 3-year CIR of 40% for VAF greater than 33% versus 7% for VAF less than 33%. When adjusted for site of KIT mutation and end-of-induction measurable residual disease, VAF greater than 33% correlated with poor OS (hazard ratio [HR]: 4.4 [95% CI: 1.2-17.2], p = .034). Exon 17 KIT mutations serve as an important predictor of relapse in RUNX1::RUNX1T1 pediatric AML. In addition, a high KIT VAF may predict poor outcomes in these patients. These results emphasize the need to incorporate KIT mutational analysis into risk stratification for pediatric CBF-AML.


Assessing the transferability of common top-down and bottom-up coarse-grained molecular models for molecular mixtures.
The performance of three methods for developing new coarse-grained models for molecular simulation is critically assessed. Two bottom-up approaches are employed: iterative Boltzmann inversion (IBI) and the multiscale coarse-graining method (MS-CG), using an atomistic n-octane-benzene reference system. Results are compared to a top-down coarse-graining approach employing the SAFT-γ Mie equation of state. The performance of each methodology is assessed against the twin criteria of local structure prediction and accurate free energy representation. In addition, the transferability of the generated potentials is compared across state points. We examine the extent to which the IBI methodology can be improved by using a multi-reference approach (MS-IBI), and demonstrate how a pressure correction can be employed to improve the results for the MS-CG approach. Additionally, we look at the effect of including angle-terms in the SAFT-γ Mie model. Finally, we discuss in detail the strengths and weaknesses of each method and suggest possible ways forward for coarse-graining, which may eventually address the problems of structure prediction, thermodynamic consistency and improved transferability within a single model.


Defining future directions in spinal cord tumor research: proceedings from the National Institutes of Health workshop.
The relative rarity of spinal cord tumors has hampered the study of these uncommon nervous system malignancies. Consequently, the understanding of the fundamental biology and optimal treatment of spinal cord tumors is limited, and these cancers continue to inflict considerable morbidity and mortality in children and adults. As a first step to improving the outcome of patients affected with spinal cord tumors, the National Institutes of Health Office of Rare Diseases Research in cooperation with the National Cancer Institute and the National Institute of Neurological Disorders and Stroke convened a workshop to discuss the current status of research and clinical management of these tumors. The overall goal of this meeting was to initiate a process that would eventually translate fundamental basic science research into improved clinical care for this group of patients. Investigational priorities for each of these areas were established, and the opportunities for future multidisciplinary research collaborations were identified.


Screening for oral premalignancy and cancer: what platform and which biomarkers?
This perspective on the report by Pattani et al. in this issue of the journal (beginning on page 1093) examines the utility of detecting hypermethylation of the candidate tumor suppressor genes endothelin receptor type B (EDNRB) and kinesin family member 1A (KIF1A) as a means of oral cancer or premalignancy screening. The data discussed here raise the possibility that saliva-based hypermethylation studies may hold promise as a cancer screening platform. This perspective also discusses some of the challenges and current limitations of developing biomarkers to screen not only for oral premalignancy and early cancer but for human papillomavirus-related oropharyngeal neoplasia as well.


What Does It Take to Become an Academic Plastic Surgeon in Canada: Hiring Trends Over the Last 50 Years.
Objective: Academic plastic surgery positions have become highly competitive secondary to delayed retirement, stagnant hospital funding, and an increasing number of plastic surgery graduates. Little information is available to help residents navigate this challenging landscape. Our objectives were to evaluate the training backgrounds of all Canadian academic plastic surgeons and to develop recommendations for residents interested in an academic career. Methods: All Canadian academic plastic surgeons were included. Training histories were obtained from institutions' websites. Surgeons were subsequently emailed to confirm this information and complete missing details. Multivariate regressions were designed to analyze the effect of gender and FRCSC year on graduate and fellowship training and time to first academic position. Results: Training information was available for 196 surgeons (22% female), with a 56% email response rate; 91% of surgeons completed residency in Canada; 94% completed fellowship training, while 43% held graduate degrees; 74% were employed where they previously trained. Female gender significantly lengthened the time from graduation to first academic job, despite equal qualification. Younger surgeons were more likely to hold graduate degrees (P < .01). Conclusions: We identified objective data that correlate with being hired at an academic centre, including training at the same institution, obtaining a graduate degree during residency, and pursuing fellowship training. In addition, we demonstrated that women take significantly longer to acquire academic positions (P < .01), despite equal qualification. Trainees should consider these patterns when planning their careers. Future research should explore gender-based discrepancies in hiring practices.


Antihistamines for the common cold.
Although antihistamines are prescribed in large quantities for the common cold, there is little evidence to whether these drugs are effective. To assess in patients with a common cold the effects of antihistamines in alleviating nasal symptoms, or in shortening of illness duration. We searched the Cochrane Acute Respiratory Infections Group Specialized Register and EMBASE up to December 2002; Cochrane Central Register of Controlled Trials (CENTRAL) and MEDLINE up to February 2003. We also followed up references in identified papers. We appealed for further articles at a major international conference on Acute Respiratory Infections (1997). We corresponded with experts and got in touch with pharmaceutical companies. Randomised, placebo-controlled trials on treatment of common cold with antihistamines, used either singly or in combination, in adults or children. Data were extracted by two reviewers and authors were contacted for further data. Trials were subdivided into monotherapy and combination therapy. Data on general recovery, nasal obstruction, rhinorrhea, sneezing, and side-effects were extracted and summarized in a systematic review. We included thirty two papers describing 35 comparisons; 22 trials studied monotherapy, 13 trials a combination of antihistamines with other medication. A total of 8930 people suffering from the common cold were included. There were large differences in study designs, participants, interventions, and outcomes. There was no evidence of any clinically significant effect - in children or in adults - on general recovery of antihistamines in monotherapy. First generation - but not non-sedating - antihistamines have a small effect on rhinorrhea and sneezing. In trials with first generation antihistamines the incidence of side effects (especially sedation) is significantly higher with active treatment. Two trials, studying a combination of antihistamines with decongestives in small children, both failed to show any effect. Of the eleven trials on older children and adults, the majority show an effect on general recovery and on nasal symptom severity. Antihistamines in monotherapy - in children as well as in adults - do not alleviate to a clinical extend nasal congestion, rhinorrhoea and sneezing, or subjective improvement of the common cold. First generation antihistamines also cause more side-effects than placebo, in particular they increase sedation in cold sufferers. Combinations of antihistamines with decongestives are not effective in small children. In older children and adults most trials show a beneficial effect on general recovery as well as on nasal symptoms. It is however not clear whether these effects are clinically significant.


Addressing Community Health Needs through the Development of a Student-Led Community Health Fair.
This article was migrated. The article was marked as recommended. Based on a specific community benefit analysis of Greenville, South Carolina, we identified the Dunean community with its increased prevalence of health inequities with respect to access to health care, poverty burden, and disease mortality on a county, state, and national level. The Dunean community's data reflect poorer health outcomes in terms of disease and unhealthy lifestyle as well as inadequate access to medical resources compared to other communities in South Carolina. Students, residents, attendings, faculty, and staff from the University of South Carolina School of Medicine Greenville (UofSC SOMG) formed a task force to engage the community and combat the root causes of diseases. This task force built partnerships with community leaders to create Root Cause, a monthly health event designed to bring community members to a unified space, share a free community dinner, and provide a wide range of health and wellness resources to educate and inspire them to make healthy lifestyle choices. Outcomes: This report describes the formation of the community engagement task force and execution of Root Cause. In five Root Cause events, we partnered with 36 community agencies and our academic health center partners who shared their resources, served 207 Dunean neighborhood members, and facilitated 1,237 total interactions between community members and partners. Under the Root Cause model, medical students and neighborhood partners have initiated a trusted, bidirectional dialogue to determine their specific needs with the desire to positively transform the health and wellness of the Dunean community. Our data suggests that based on our efforts, the neighborhood of Dunean, SC increased community cohesiveness and improved perceptions of access to health care. Additionally, participating medical students advanced their understanding of social health and economic challenges which helped to facilitate their development along the active citizen continuum, as well as increase empathy for their future patients in the local community.


Application of a robotic telemanipulation system in stereotactic surgery.
To assess the clinical usefulness, accuracy, and safety of telemanipulation for frameless stereotactic surgery using the CAS-BH5 robot system, we prospectively evaluated 10 patients (age: 5-79 years; mean: 44 years) who underwent telemanipulation frameless stereotactic operations from September to December 2005. The CAS-BH5 robot system consists of three main parts: a planning subsystem, a surgical localization subsystem, and a telemanipulation subsystem. Specifically, CAS-BH5 is capable of network communication, video transmission, graphic simulation and human-machine interaction, and thus facilitates remote planning and transmission of neuronavigation data, monitoring and manipulating. Telemanipulation was performed via a digital data network with a speed of 2,000 kilobytes per second by a neurosurgeon in Beijing while the patients were located in Yan'an, 1,300 km away. Remote fiducial registration was performed with a mean accuracy of 1.05 mm and the standard difference between the planned and actual trajectory was 0.13 mm. The mean time from fiducial registration to closure was 30.2 +/- 1.66 min. At 12-month follow-up, 90% of patients had improved neurologically. There were no complications. This preliminary data indicates that telemanipulation in frameless stereotactic surgeries is feasible, reliable and safe. In the future, we believe that telemanipulation will facilitate collaboration between surgeons, enhance training, allow for sharing of resources, and have wide applications in the field of neurosurgery.


[Evolution of serum lipids during the first hours of an acute myocardial infarction (author's transl)].
The evolution of serum lipids is investigated in 26 patients admitted in a coronary care unit less than 9 hours after the onset of an acute myocardial infarction. Blood samplings are made every 3 hours until the 15th hour after the onset of the infarction. The decrease of total serum lipids concentration is significant at the 9th hour (3rd hour : 8,45 +/- 0,58 g/l ; 9th hour : 7,74 +/- 0,36 g/l) ; that of triglycerides, at the 6th hour (3rd hour : 1,33 +/- 0,17 g/l ; 6th hour : 1,18 +/- 0,21 g/l) ; that of cholesterol is more delayed, at the 9th hour (3rd hour : 2,98 +/- 0,18 g/l ; 9th hour : 2,83 +/- 0,11 g/l). The proportion of alphalipoproteins increases, that of prebetalipoproteins decreases, that of betalipoproteins remains unchanged. There is no difference between the patients who received heparin and those who did not. Total lipids and triglycerides concentrations are lower in the group of patients with complications than in the group without complications.


Differences in Occupational Burnout Among Primary Care Professionals.
Occupational burnout is a major concern for personal well-being and patient care. We examined burnout among primary care providers (PCPs), medical residents, behavioral health providers (BHPs), nurses, and other clinical and nonclinical primary care team members. This was a cross-sectional study, nested within a larger randomized trial. Participants completed a validated 9-item burnout measure with 3 domains: depersonalization, emotional exhaustion, and personal accomplishment. Multivariable multilevel linear regression with a random intercept for each practice was used to determine mean differences in burnout across professional roles. Overall burnout rates varied by professional role: PCPs 70%, medical residents 89%, BHPs 59%, nurses 66%, other clinicians 68%, and nonclinical professionals 70%. Compared with nonclinical professionals, residents experienced more burnout in more domains, followed by PCPs. PCPs, residents, and nurses reported significantly worse depersonalization and exhaustion scores. Nonclinical professionals had worse accomplishment scores than all clinical professionals except for residents. This study revealed moderate-to-high levels of burnout among primary care professionals. Clinicians may be experiencing aspects of burnout more intensely than their nonclinical colleagues, and this may be most true for residents and PCPs. Based on these variations, interventions to mitigate burnout may need to be tailored by professional role.


The Duality of Oral Sex for Men Who Have Sex with Men: An Examination Into the Increase of Sexually Transmitted Infections Amid the Age of HIV Prevention.
Several studies suggest that the increase in sexually transmitted infections (STIs) among men who have sex with men (MSM) could be due, in part, to human immunodeficiency virus (HIV) risk reduction strategies, which include engaging in oral sex over anal sex. The aims of this study were to evaluate oral sex behaviors and STI diagnoses and to investigate the potential dual role of oral sex as being protective for HIV, yet perpetuating STI transmission among MSM. We analyzed records from 871 MSM presenting to the Rhode Island STI Clinic between 2012 and 2015. We compared outcomes in men engaging in two HIV protective oral sex behaviors: (1) HIV/STI outcomes by men engaging only in oral sex versus those that did not, and (2) HIV/STI outcomes by men engaging in condomless oral sex with 100% condom use for all other sex acts versus those that did not. Men engaging in HIV protective oral sex behaviors were more likely to be HIV negative compared to men not engaging in them (99% vs. 93%, p < 0.01). In contrast, there was no significant difference in STI diagnoses between those that engaged in HIV protective oral sex behaviors and those that did not. The findings provide evidence to support the unique duality of oral sex: decreased risk for HIV and perpetuation of STI risk. Promotion of routine STI testing, including extragenital sites, is critical to address STI prevention among MSM. In the age of HIV prevention, addressing the ambiguous risks of discrete sex acts would be beneficial for both HIV and STI prevention education for MSM.


Is distortion correction necessary for digital subtraction angiography in the Gamma Knife treatment of intra-cranial arteriovenous malformations?
The published literature maintains that distortion correction of digital subtraction angiography is essential for accurate definition of the arteriovenous malformation (AVM) nidus when performing Gamma Knife radiosurgery. However, as Gamma Knife treatments usually involve small volume targets, which are purposely positioned as close to the centre of the frame as possible, we postulated that the angiographic target images should be minimally distorted. Thus, the use of uncorrected images would not significantly compromise the accuracy of the treatment. We investigated this postulate. Patient and phantom images with and without distortion correction were assessed. The errors measured in the angiograms were compared to those of MR and CT images of the same subjects. Any errors less than 0.5 mm in any modality were accepted as intrinsic. All errors measured in the study were sub-millimetre. In the patient group, errors with distortion correction averaged 0.13 mm (range 0-0.3 mm). Without distortion correction, errors averaged 0.34 mm (range 0.1-0.6 mm). By comparison, the average error for intracranial MR imaging was 0.3 mm, with a maximum of 0.6 mm. For small, intracranial AVMs distortion correction did not significantly reduce the errors associated with the definition of the stereotactic co-ordinates. We suggest that prior to the purchase of this software, similar in-house testing should be performed.


Characterization of copy number-stable regions in the human genome.
In the past few years the number of copy number variants (CNVs) identified in the human genome has increased significantly, but our understanding of the functional impact of CNVs is still limited. Clinically significant variations cannot easily be distinguished from benign, complicating interpretation of patient data. Multiple studies have focused on analysis of regions that vary in copy number in specific disorders. Here we use the opposite strategy and focus our analysis on regions that never seem to vary in the general population, hypothesizing that these are copy number stable because variations within them are deleterious. Our results show that copy number stable regions are characterized by correlation with a number of genomic features, allowing us to define a list of genomic regions that are dosage sensitive in humans. We find that these dosage-sensitive regions show significant overlap with de novo CNVs identified in patients with intellectual disability or autism. There is also a significant association between copy number stable regions and rare inherited variants in autism patients, but not in controls. Based on this predictive power, we propose that copy number stable regions can be used to complement maps of known CNVs to facilitate interpretation of patient data.


Inhibition of fatty acid incorporation into adipose tissue triglycerides in Ehrlich ascites tumor-bearing mice.
Using a recently developed technique of direct tracer injection into selective adipose tissue sites (Baker et al., Mech. Ageing Dev., 27: 295-313, 1984), we have studied the esterification of free fatty acids (FFA) to triglyceride fatty acids in the epididymal fat pads of normal and Ehrlich ascites carcinoma-bearing mice. We have tested the hypothesis that, during Ehrlich ascites carcinoma growth, a defect develops, resulting in the inhibition of the esterification and incorporation of FFA into adipose tissue diglyceride and triglyceride fatty acids. Our technique allowed the measurement of the disappearance of [1-14C]palmitic acid as FFA and its incorporation into di- and triglyceride fatty acids over 1 h. Multicompartmental analysis was used to compute the fractional rates of esterification and turnover. Using measured FFA pool sizes and assuming near-steady-state conditions, we estimated the transport rates (mass/time) of fatty acid esterification and turnover. Our results indicate that, compared to controls (normal mice), the epididymal fat pads of mice bearing early (5-day) and advanced (9-day) Ehrlich ascites carcinoma, respectively, show: 65% and near complete (congruent to 99%) decreases in the fractional rates of FFA esterification; about 2- and 24-fold increases in the FFA pool sizes; and 40% and 70% decreases in the transport rates of esterification.


Carbohydrate metabolism in women with a twin pregnancy.
Carbohydrate metabolism was evaluated in 24 women with a twin pregnancy and 24 women with a singleton pregnancy. The groups were of similar age, parity, weight, and gestational age. In each woman an intravenous glucose tolerance test was done using a 25-g glucose load in the last half of gestation. Both blood glucose and plasma insulin levels were measured and statistically compared. The plasma human placental lactogen levels were significantly higher in the women with the twin gestation (7.3 +/- 0.7 versus 4.7 +/- 0.3 microgram/ml). Although the glucose disappearance rates (K) were not different, there was a significantly lower fasting as well as 5- and 15-minute blood glucose value in the twin pregnancy group. There was also a significantly lower 15-minute insulin level in the twin group. The importance of these findings to the clinical management of twin pregnancies and to the understanding of the metabolic changes in pregnancy is discussed.


Findings in patients from Benin with osteomyelitis and polymerase chain reaction-confirmed Mycobacterium ulcerans infection.
Mycobacterium ulcerans is known to cause Buruli ulcer (BU), a necrotizing skin disease leading to extensive cutaneous and subcutaneous destruction and functional limitations. However, M. ulcerans infections are not limited to skin, and osteomyelitis, still poorly described in the literature, occurs in numerous young patients in Africa. In a retrospective matched case-control study conducted in a highly endemic area in Benin, we analyzed demographic, clinical, biological, and radiological features in all patients with M. ulcerans infections with bone involvement, identified from a cohort of 1257 patients with polymerase chain reaction-proved M. ulcerans infections. The 81 patients studied had a median age of 11 years (interquartile range, 7-16 years) and were predominantly male (male-female ratio, 2:1). Osteomyelitis was observed beneath active BU lesions (60.5%) or at a distance from active or apparently healed BU lesions (14.8%) but also in patients without a history of BU skin lesions (24.7%). These lesions had an insidious course, with nonspecific clinical findings leading to delayed diagnosis. A comparison with findings in 243 age- and sex-matched patients with BU without osteomyelitis showed that case patients were less likely to have received BCG immunization than controls (33.3% vs 52.7%; P = .01). They were also at higher risk of longer hospital stay (118 vs 69 days; P = .001), surgery (92.6% vs 63.0%; P = .001), and long-term crippling sequelae (55.6% vs 15.2%; P < .001). This study highlighted the difficulties associated with diagnosis of M. ulcerans osteomyelitis, with one-fourth of patients having no apparent history of BU skin lesions, including during the current course of illness. Delays in treatment contributed to the high proportion (55.6%) of patients with crippling sequelae.


In defence of a single body of clinical and public health, medical ethics.
Since some form of dual clinical/public health practice is desirable, this paper explains why their ethics should be combined to influence medical practice and explores a way to achieve that. In our attempt to merge clinical and public health ethics, we empirically compared the individual and collective health consequences of two illustrative lists of medical and public health ethical tenets and discussed their reciprocal relevance to praxis. The studied codes share four principles, namely, 1. respect for individual/collective rights and the patient's autonomy; 2. cultural respect and treatment that upholds the patient's dignity; 3. honestly informed consent; and 4. confidentiality of information. However, they also shed light on the strengths and deficiencies of each other's tenets. Designing a combined clinical and public health code requires fleshing out three similar principles, namely, beneficence, medical and public health engagement in favour of health equality, and community and individual participation; and adopting three stand-alone principles, namely, professional excellence, non-maleficence, and scientific excellence. Finally, we suggest that eco-biopsychosocial and patient-centred care delivery and dual clinical/public health practice should become a doctor's moral obligation. We propose to call ethics based on non-maleficence, beneficence, autonomy, and justice - the values upon which, according to Pellegrino and Thomasma, the others are grounded and that physicians and ethicists use to resolve ethical dilemmas - "neo-Hippocratic". The neo- prefix is justified by the adjunct of a distributive dimension (justice) to traditional Hippocratic ethics. Ethical codes ought to be constantly updated. The above values do not escape the rule. We have formulated them to feed discussions in health services and medical associations. Not only are these values fragmentary and in progress, but they have no universal ambition: they are applicable to the dilemmas of modern Western medicine only, not Ayurvedic or Shamanic medicine, because each professional culture has its own philosophical rationale. Efforts to combine clinical and public health ethics whilst resolving medical dilemmas can reasonably be expected to call upon the physician's professional identity because they are intellectual challenges to be associated with case management.


Campylobacter pyloridis gastritis: the past, the present, and speculations about the future.
Campylobacter pyloridis infection of the stomach has been associated with gastric ulcer, duodenal ulcer, nonulcer dyspepsia, and gastritis. The etiological role of C. pyloridis in most of those conditions remains unclear. We reviewed what is known about C. pyloridis infections in man. Considerable clinical data on C. pyloridis infections was available in older literature concerning gastritis and gastric urease. C. pyloridis causes a form of type B gastritis. In some individuals the acute infection is associated with abdominal pain and transient hypochlorhydria. C. pyloridis infection is difficult to eradicate with current therapies. The mechanisms by which C. pyloridis infection may lead to development of peptic ulcers, nonulcer dyspepsia, or atrophic gastritis are discussed. Recent technological advances, such as the 13C-urea breath test, provide rapid noninvasive methods of identifying active C. pyloridis infection. These methods will permit the rapid execution of definitive investigations of the epidemiology, transmission patterns, and possible reservoirs of C. pyloridis infection and will delineate the spectrum of C. pyloridis-associated disorders.


The role of serum biomarkers in predicting fibrosis progression in pediatric and adult hepatitis C virus chronic infection.
Liver biopsy represents the gold standard for damage evaluation, but noninvasive serum markers that mirror liver fibrosis progression are actual goals both in adults and especially in children. The aim was to determine specific serum markers that correlate with liver fibrosis progression during chronic HCV infection. Liver biopsies and concomitant serum samples from 22 pediatric and 22 adult HCV patients were analyzed. Histological parameters were evaluated. On serum TGF-ß1, tissue inhibitor of matrix metalloprotein inhibitor-1 (TIMP-1), hyaluronic acid (HA) and aminoterminal peptide of procollagen type III (PIIINP) were tested. Significant fibrosis (F≥2) and advanced fibrosis (F≥3) represented 64% and 20%, respectively in children; while 54% F≥2 and 23% F≥3 in adults. Hyaluronic acid (p = 0.011) and PIIINP (p = 0.016) were related to worse fibrosis stages only in adults, along with TIMP-1 (p = 0.039) just in children; but TGF-ß1 was associated with mild fibrosis (p = 0.022) in adults. The AUROC of TIMP-1 in children to discriminate advanced fibrosis was 0.800 (95%IC 0.598-0.932). In adults, the best AUROCs were that of HA, PIIINP and TGF-ß1 [0.929 (IC95% 0.736-0.994), 0.894 (IC95% 0.689-0.984) and 0.835 (IC95% 0.617-0.957)], respectively. In children, according to the cut off (165.7 ng/mL) value for TIMP-1, biopsies could have been avoided in 72% (18/25). Considering the cut off for HA (109.7 ng/mL), PIIINP (9.1 µg/L), and TGF-ß1 (10,848.3 pg/mL), biopsies could have been avoided in 87% (19/22) of adult patients by using HA and 73% (16/22) using PIIINP or TGF-ß1. In adults given the diagnostic accuracy of HA, PIIINP, TGF-ß1, their combination may provide a potential useful tool to assess liver fibrosis. This first pediatric study suggests that TIMP-1 is clinically useful for predicting liver fibrosis in HCV patients.


Carotid artery disease in young adults.
The angiographic and clinical findings in 17 young adults (14-39 years) with stenosis and/or occlusion of at least one of the common or internal carotid arteries were reviewed to determine the spectrum of carotid artery disease in this age group. Five aetiological groups were identified: atherosclerosis (3 patients), arteritis (4), non-inflammatory arteriopathies (6), compression by neoplasm (1), and occlusions of undetermined cause (3). Over a mean follow-up time of 19.7 months, 2 patients died (12.5%) and further cerebrovascular events occurred in 4 others (25%). We conclude that carotid angiography plays a meaningful role in providing a definitive diagnosis in young adults with carotid artery disease and thus may assist with therapy and prognosis.


Development and validation of the mini-osteoporosis quality of life questionnaire (OQLQ) in osteoporotic women with back pain due to vertebral fractures. Osteoporosis Quality of Life Study Group.
The objective of the study was to evaluate a shortened osteoporosis quality of life questionnaire (OQLQ) in osteoporotic women with back pain due to vertebral fractures. From the longer 30-item OQLQ (four to nine items per domain) we created the mini-OQLQ by choosing the two items with the highest impact in each of five domains (symptoms, physical function, activities of daily living, emotional function, leisure). We administered the OQLQ, the Sickness Impact Profile, the SF-36 and the Brief Pain Index to patients at baseline, after 2 weeks and after 6 months. The intraclass correlations between baseline and the 2-week follow-up for the five mini-OQLQ domains ranged from 0.72 to 0.86. Cross-sectional correlations between the domains of the mini-OQLQ and other health instruments were moderate to large (0.35-0.80) and greater than predicted. The mini-OQLQ items showed moderate to large correlations with items omitted from the shortened questionnaire (0. 44-0.88). Correlations between the OQLQ domains and the other three instruments were greater than those of the mini-OQLQ, and partial correlations between OQLQ items omitted from the mini-OQLQ and the other three instruments after considering mini-OQLQ items were substantial (0.19-0.71) and statistically significant. Sample sizes of less than 200 per group should be required to detect minimally important differences in parallel-group clinical trials. Longitudinal correlations between the mini-OQLQ and the other measures were often significant but generally lower than predicted (0.10-0.49). The partial correlations revealed that the omitted items explained a significant portion of the longitudinal variance in each domain. We conclude that in a selected group of patients with back pain caused by vertebral fractures, the mini-OQLQ demonstrated good discriminative and adequate evaluative properties. The mini-questionnaire should be useful in clinical settings.


Autophagy protects the retina from light-induced degeneration.
Autophagy is a conserved feature of lysosome-mediated intracellular degradation. Dysregulated autophagy is implicated as a contributor in neurodegenerative diseases; however, the role of autophagy in retinal degeneration remains largely unknown. Here, we report that the photo-activated visual chromophore, all-trans-retinal, modulated autophagosome formation in ARPE19 retinal cells. Increased formation of autophagosomes in these cells was observed when incubated with 2.5 μM all-trans-retinal, a condition that did not cause cell death after 24 h in culture. However, autophagosome formation was decreased at concentrations, which caused cell death. Increased expression of activating transcription factor 4 (Atf4), which indicates the activation of oxidative stress, was recorded in response to light illumination in retinas of Abca4(-/-)Rdh8(-/-) mice, which showed delayed clearance of all-trans-retinal after light exposure. Expression of autophagosome marker LC3B-II and mitochondria-specific autophagy, mitophagy, regulator Park2, were significantly increased in the retinas of Abca4(-/-)Rdh8(-/-) mice after light exposure, suggesting involvement of autophagy and mitophagy in the pathogenesis of light-induced retinal degeneration. Deletion of essential genes required for autophagy, including Beclin1 systemically or Atg7 in only rod photoreceptors resulted in increased susceptibility to light-induced retinal damage. Increased photoreceptor cell death was observed when retinas lacking the rod photoreceptor-specific Atg7 gene were coincubated with 20 μM all-trans-retinal. Park2(-/-) mice also displayed light-induced retinal degeneration. Ultra-structural analyses showed mitochondrial and endoplasmic reticulum impairment in retinas of these model animals after light exposure. Taken together, these observations provide novel evidence implicating an important role of autophagy and mitophagy in protecting the retina from all-trans-retinal- and light-induced degeneration.


[The effect of tropoxin on the neurogenic and vasogenic inflammation of the dura mater vessels].
Subchronic administration of tropoxin (in doses of 7.5 and 10 mg/kg) caused dose-dependent blocking of 131I-albumin plasma transudation from the dura mater vessels, induced by electrical stimulation of the trigeminal ganglion and intravenous infusion of the agonist of 5-HT2B/2C receptors metachlorophenylpiperazine. The antimigraine agent metisergid produced a similar effect. A single injection of metisergid and tropoxin did not block albumin transudation. A 3 mg/kg dose of mianserin prevented the blocking effect of tropoxin and metisergid on plasma exudation into the dura mater. It is suggested that the mechanism of the tropoxin antimigraine effect is realized through the presynaptic 5-HT1 receptors of afferent endings of the trigeminal nerve and the postsynaptic 5-HT2B/2C receptors of the dura mater vessels.


Relative magnitude of tonic and phasic synaptic excitation of medullary inspiratory neurons in dogs.
The relative contribution of phasic and tonic excitatory synaptic drives to the augmenting discharge patterns of inspiratory (I) neurons within the ventral respiratory group (VRG) was studied in anesthetized, ventilated, paralyzed, and vagotomized dogs. Multibarrel micropipettes were used to record simultaneously single-unit neuronal activity and pressure microejected antagonists of GABAergic, glycinergic, N-methyl-D-aspartate (NMDA) and non-NMDA glutamatergic, and cholinergic receptors. The discharge patterns were quantified via cycle-trigger histograms. The findings suggest that two-thirds of the excitatory drive to caudal VRG I neurons is tonic and mediated by NMDA receptors and the other third is ramp-like phasic and mediated by non-NMDA receptors. Cholinergic receptors do not appear to be involved. The silent expiratory phase is produced by phasic inhibition of the tonic activity, and approximately 80% of this inhibition is mediated by gamma-aminobutyric acid receptors (GABA(A)) and approximately 20% by glycine receptors. Phasic I inhibition by the I decrementing neurons does not appear to contribute to the predominantly step-ramp patterns of these I neurons. However, this decrementing inhibition may be very prominent in controlling the rate of augmentation in late-onset I neurons and those with ramp patterns lacking the step component.