Saturated fatty acids and coronary heart disease risk: the debate goes on.
Recently published meta-analyses of cohort studies and randomized controlled trials (RCTs) have challenged the link between saturated fatty acid (SFA) intake and coronary heart disease (CHD) risk. This review considers the outcome of these studies in the context of other evidence. Recent meta-analyses of cohort studies suggest that reducing SFA intakes has little impact on CHD risk when replaced by carbohydrates. The evidence for benefits on CHD risk of replacing SFA with unsaturated fatty acids in cohort studies is stronger and is also supported by data from a recent Cochrane analysis of RCTs of dietary SFA reduction and CHD risk. This review highlights the challenges of cohort studies involving diet because of the changing patterns of dietary behaviour and other multifactorial risk factors. The studies included are normally conducted over many years and are often dependent on a single measurement of dietary intake. The link between SFA intake, plasma cholesterol, and CHD risk is based on a broad range of evidence including mechanistic studies, RCTs of surrogate end points and clinical outcomes, as well as multinational population comparisons. Public health nutrition policy should continue to take into account the totality of evidence with recognition of the limitations of dietary cohort studies.


Safety of immediate treadmill testing in selected emergency department patients with chest pain: a preliminary report.
To determine the feasibility and safety of an immediate, symptom-limited, treadmill test on selected emergency department (ED) patients, a convenience sample of 28 patients underwent an exercise treadmill test (ETT) within the first several hours after hospital arrival using the modified Bruce protocol. Patients were included in the study if they presented with otherwise unexplained chest pain consistent with (but not characteristic for) angina pectoris and had a normal electrocardiogram. A negative ETT was seen in 23 of 28 patients, and five of 28 patients had a positive ETT. No patients had serial enzyme or electrocardiogram evolution suggestive of myocardial ischemia, and all patients with a negative ETT were discharged after a full inpatient evaluation designed to rule out unstable coronary disease. At a mean follow-up period of 6.1 months there has been no cardiac morbidity or mortality in the patients with negative ETTs. It was concluded that early ETTS of selected ED patients with chest pain is safe, and an exercise test administered during the ED visit which is negative can preclude unnecessary hospitalization.


Programmed cell death and apoptosis: origins of the theory.
Interest in the study of apoptosis grew with the recognition that it is a highly regulated process. Such a change in attitude allowed the intellectual and technical breakthroughs that led to the explosive development of this subject.


The need for health education among patients with psoriasis.
Psoriasis is one of the most common skin diseases with a chronic and recurrent course. The disease requires systematic treatment and appropriate care. Therefore, in the care of the patient with psoriasis, there must also be place for health education. The aim of this study was to determine the need for health education among patients with psoriasis by determining knowledge gaps occurring most frequently. Materials for this study were collected using an anonymous questionnaire created on the basis of the Psoriasis Disability Index (PDI). The study group included 149 patients with psoriasis. It has been proven in previous studies that patients with psoriasis see the need to improve their knowledge of the disease and self-care methods, and the greatest deficit of their education refers to the factors activating the process of the disease and to the methods of prophylactics in psoriasis.


Expression of c-Myc, neurofibromatosis Type 2, somatostatin receptor 2 and erb-B2 in human meningiomas: relation to grades or histotypes.
Meningiomas, which originate from arachnoid cells, represent one of the largest subgroups of intracranial tumors. They are generally benign, but can progress to malignancy. The aim of our study was to determine the expression of 4 genes, c-Myc, neurofibromatosis Type 2 (NF2), somatostatin receptor isoform 2 (sst2) and erb-B2, that have been associated with tumorogenesis or, possibly, with aggressive behavior or recurrence of meningiomas. We measured levels of mRNAs coding for these genes by qRT-PCR in 51 cases and levels ofc-Myc protooncogene and sst2 protein by immunohistochemistry in 26 cases of meningiomas of various grades and histotypes. C-Myc mRNA and protein levels were not grade-related, but validated subdivision of the 36 benign meningiomas into two groups, Groups IA and IB, based on histological and clinical features (Ki-67-proliferative index, absence or presence of mitoses, rate of recurrence and incidence of perilesional edema). In addition to histopathological grading, c-Myc expression may be useful in predicting tumor recurrence in patients with low-grade meningiomas. NF2 mRNA levels and sst2 mRNA and receptor levels were not grade-related, but were histotype-related, with significantly higher levels in the meningothelial subtype than in the fibroblastic subtype. Erb-B2 mRNA levels were not grade- or histotype-related. Furthermore, the high expression of sst2 in meningothelial meningioma suggests the possibility of a different tumorigenesis process in this meningioma subtype and may open perspectives for the diagnosis and therapy of this subtype using somatostatin as an antiproliferative agent.


Identification of radiation-responsive genes in vitro using a gene trap strategy predicts for modulation of expression by radiation in vivo.
A large number of genes are known to be responsive to ionizing radiation, and there is strong evidence for the existence of inducible radiation resistance in mammalian cells. We have developed a gene trap insertional mutagenesis strategy to identify novel genes involved in responses to radiation. Using this approach, we have isolated four gene-trap integrations in embryonic stem cells. In three cases (9A, 3E and 9H) the trapped genes are radiation-inducible, and in one (7D) the gene is down-regulated. Sequence analysis of fusion transcripts from three of the integrations indicate one novel gene (3E), the mouse homologue (9A) of a known but uncharacterized human gene that encodes a protein with significant homology to several GTPase-activating proteins and a murine locus, Mym (9H). The embryonic stem cell clone with the 9A insertion was introduced into the mouse germline, and the in vivo expression pattern of 9A was studied in detail. A unique, spatially restricted pattern of expression in embryos and adult animals was observed. There is tissue-specific in vivo induction of the 9A gene in adult mice by radiation. This study demonstrates the potential of the gene trap approach for the identification and functional analysis of novel radiation-regulated genes. Similar strategies may facilitate the discovery and characterization of genes involved in other cellular stress responses.


Cyclic nucleotides, adenylate cyclase, and cyclic AMP phosphodiesterase in mammary glands from pregnant and lactating mice.
In the mammary glands of mice, levels of cyclic AMP increased during pregnancy and then fell precipitously following parturition. In contrast, levels of cyclic GMP fell during the gestation period and then rose rapidly during the early days of lactation. Adenylate cyclase and cyclic AMP hohsphodiesterase activities were elevated during the pregnancy and lactation periods.


Digital subtraction angiography in selection of the vascularized greater trochanter bone grafting for treatment of osteonecrosis of femoral head.
The aim of this report is to present our experience on the use of the digital subtraction angiography (DSA) in selection of the vascularized greater trochanter bone grafting for the treatment of the osteonecrosis of femoral head (ONFH) in early stages. Between January 2005 and June 2007, DSA was used to evaluate the blood perfusion of the early stages ONFH in 32 patients (45 hips). There were 18 males and 14 females with an average age of 30 years old. Twenty-one hips were in ARCO stage I, and 24 in ARCO stage II. The arterial blood supply insufficiency was found in 22 hips by DSA, and the venous stasis in 23 hips. The hips with artery blood supply insufficiency received the vascularized greater trochanter bone grafting, and the hips with the venous stasis received the core decompression. All of patients were followed-up with an average of 4.8 years (ranging 2.4-6.6 years). The preoperative Harris Hip score (HHS) in the patients with arterial blood supply insufficiency was 48.18 ± 7.81 and the postoperative HHS was 93.27 ± 3.03. The preoperative HHS in the patients with venous stasis was 44.04 ± 6.40, and the postoperative HHS 92.65 ± .93. The postoperative DSA showed an improved perfusion of the femoral head in 44 hips. Our experience showed that DSA would help to select the appropriate procedure for treatment of ONFH in the early stage.


Tool to assess the cost and quality benefits of nursing innovation.
Understanding the economic value of nursing services in a time of unprecedented public sector cuts is a challenge. The economic assessment tool (EAT) (RCN 2011) has been designed by the authors of the article for this purpose and generates return on investment dividends for nursing innovations and services. The EAT, which is built on the discipline of improvement and uses many of its tools and techniques, involves four stages: mapping, costing, calculating and reporting. The nursing profession systematically captures a range of clinical data as part of routine care to which monetary values can be assigned. The EAT exploits these data and provides the profession with the economic evidence it might need to sustain quality nursing services in financially uncertain times.


The charge ratio between O and N on amide bonds: a new approach to the mobile proton model.
The influence of charge distribution on the cleavage of the peptides was investigated by fragmentation efficiency curves and quantum chemical calculations in order to clarify the fragmentation mechanism in this paper. The peptide Arg-Gly-Asp-Cys (RGDC) was oxidized to change the charge distribution, but its main sequence was retained. Under this study, it was illustrated that the fragmentation of the peptide RGDC became easier with each addition of an O atom to the Cys hydrosulfide group and the relative charge ratios between O and N (QO/QN) in the amide bonds had much to do with the cleavage of the peptide RGDC. For each amide bond, the situations coincided with overall conclusion: the increase of the QO/QN values results in a higher fragmentation efficiency and vice versa. The methods which combined fragmentation efficiency curves with the charge distribution of peptides provided a way to refine the mobile proton model for peptide fragmentation and to probe the discrepant fragmentation of peptides in peptide/protein identification.


Immunological characterization of the equine airway epithelium and of a primary equine airway epithelial cell culture model.
Our understanding of innate immunity within the equine respiratory tract is limited despite growing evidence for its key role in both the immediate defense and the shaping of downstream adaptive immune responses to respiratory disease. As the first interface to undergo pathogen invasion, the respiratory epithelium is a key player in these early events and our goal was to examine the innate immune characteristics of equine respiratory epithelia and compare them to an in vitro equine respiratory epithelial cell model cultured at the air-fluid interface (AFI). Respiratory epithelial tissues, isolated epithelial cells, and four-week old cultured differentiated airway epithelial cells collected from five locations of the equine respiratory tract were examined for the expression of toll-like receptors (TLRs) and host defense peptides (HDPs) using conventional polymerase chain reaction (PCR). Cultured, differentiated, respiratory epithelial cells and freshly isolated respiratory epithelial cells were also examined for the expression of TLR3, TLR9 and major histocompatibility complex (MHC) class I and class II using fluorescence-activated cell sorting (FACS) analysis. In addition, cytokine and chemokine profiles from respiratory epithelial tissues, freshly isolated respiratory epithelial cells, and cultured, differentiated, epithelial cells from the upper respiratory tract were examined using real-time PCR. We found that respiratory epithelial tissues and isolated epithelial cells expressed TLRs 1-4 and 6-10 as well as HDPs, MxA, 2'5' OAS, β-defensin-1, and lactoferrin. In contrast, epithelial cells cultured at the AFI expressed TLRs 1-4 and 6 and 7 as well as MxA, 2'5' OAS, β-defensin-1, but had lost expression of TLRs 8-10 and lactoferrin. In addition, MHC-I and MHC-II surface expression decreased in epithelial cells cultured at the AFI compared to isolated epithelial cells whereas TLR3 and TLR9 were expressed at similar levels. Lastly, we found that equine respiratory epithelial cells express an array of pro-inflammatory, antiviral and regulatory cytokines and that after four weeks of in vitro growth conditions, equine respiratory epithelial cells cultured at the AFI retained expression of GM-CSF, IL-10, IL-8, TGF-β, TNF-α, and IL-6. In summary, we describe the development of an in vitro equine respiratory epithelial cell culture model that is morphologically similar to the equine airway epithelium and retains several key immunological properties. In the future this model will be a used to study equine respiratory viral pathogenesis and cell-to-cell interactions.


Stable dimeric magnesium(I) compounds: from chemical landmarks to versatile reagents.
The chemistry of the s-block metals is dominated by the +1 oxidation state for the Alkali metals (group 1) and the +2 oxidation state for the Alkaline Earth metals (group 2). In recent years, a series of stable dimeric magnesium(I) compounds has been prepared and their chemistry has started to develop. These complexes feature "deformable" Mg-Mg single bonds and are stabilised by sterically demanding and chelating anionic N-ligands that prevent their disproportionation. They have rapidly proven useful in organic and organometallic/inorganic reduction reactions as hydrocarbon soluble, stoichiometric, selective and safe reducing agents. The scope of this perspective focuses on stable molecular compounds of the general type LMgMgL and describes their synthesis, structures, theoretical and spectroscopic studies as well as their further chemistry. Also, comparisons are drawn with related complexes including magnesium(II) hydrides and dimeric zinc(I) compounds.


Altered cardiovascular reactivity to mental stress but not to cold pressure test in migraine.
This study assessed cardiovascular reactivity to mental stress and cold pressure test in migraineurs and controls. It compared the cardiovascular reactivity between patients with migraine with aura and patients with migraine without aura. Several studies have assessed the autonomic nervous system functioning and cardiovascular responses to stressor stimuli in migraine. Cold pressure test and sustained attention tasks are distinct forms of induced stress. It is still unknown if patients with migraine have distinct patterns of response to sustained attention tasks and cold pressure test, since no previous studies have evaluated the cardiovascular responses to these 2 distinct types of stress in the same population of migraine patients. Two distinct protocols were used to induce cardiovascular reactivity. Mental stress was induced by using a Stroop test card, a procedure involving the maintenance of the attention control. The other protocol was the cold pressure test. The blood pressure and heart rate were digitally recorded in rest and test phases. The mean elevation and the variance of blood pressure and heart rate were compared between groups. Patients with migraine had higher rest systolic blood pressure and lower heart rate induced by mental stress than controls. There were no differences between migraineurs and controls with cold pressure test. There were no differences between migraineurs with and without aura. There was a significantly different pattern of cardiovascular reactivity between migraineurs and controls with mental stress but not with cold pressure test. Distinct central nervous system structures are involved in these 2 types of stress. A distinct pattern of activation of the prefrontal cortex-periaqueductal gray matter circuit in migraine may explain a singular autonomic reactivity to mental stress in this disease.


Superficial Solitary Fibrous Tumor: A Series of 26 Cases.
While superficial (cutaneous/subcutaneous) solitary fibrous tumor (SFT) have been described, definitive diagnosis is difficult due to overlapping features with other tumors. We describe the largest series to date of superficial SFT. For inclusion, SFT had to arise in dermis or subcutis. Twenty-six cases were identified. Patients ranged from 16 to 80 years (mean, 47 y) with a marked female predominance (19 F; 7 M). Tumors involved the head (11), thigh (7), back (3), shoulder (2), upper arm (1), ankle (1), and great toe (1). Mean size was 2.9 cm (range, 1.0 to 7.0 cm). The majority (n=19) had typical histologic features (cellular SFT) with irregular fascicles of spindled cells, staghorn-like blood vessels, and variable amounts of collagen. Necrosis was evident in 3 cases (all <25%). Mitotic activity ranged from 0 to 10 mitotic figures/10 high-power fields (mean, 2 mitotic figures/10 high-power fields). Seventeen of the 18 were positive for STAT6, whereas 21/22 expressed CD34. All were low risk (23/23) by proposed criteria (Demicco and colleagues), including 2 cases with malignant histology. Three could not be risk stratified due to lack of information on tumor size. Follow-up, available on 7 cases, showed no recurrence/metastasis (mean follow-up, 100 mo; range, 2 to 241 mo). Cutaneous SFT are more common in women and most often involve the head. They are usually low risk by proposed criteria and appear to behave in an indolent manner, though larger studies are needed to confirm this. Recognition that SFT may present as a superficial mass will avoid misclassification as other CD34-positive neoplasms that frequently arise in the skin and subcutaneous tissue.


[Recent morphofunctional findings on the left atrium: their relations to thromboembolic risk].
In the past, the left atrial appendage has been considered a "useless" structure but associated to thromboembolic complications; its physiologic role is still undefined. Owing to its great distensibility, left atrial appendage positively influences atrial compliance and left ventricular performances. In addition this structure seems to play an important role in circulatory homeostasis by the release of atrial natriuretic factor in response to volume loading and atrial stretch. Transesophageal echocardiography provides a detailed anatomical characterization of this structure and, by means of Doppler flow velocities recordings, supplies relevant functional data. Despite their anatomical contiguity, the left atrium and atrial appendage result from a separate embryonic development; likewise, their function may differentiate. In the left atrial appendage a quadriphasic flow pattern has been described in subjects with sinus rhythm; however, as we reported, in some patients a more complex Doppler pattern can be observed, with an additional systolic forward flow wave which is presumably due to reflection phenomena. In patients with atrial fibrillation, an irregular flow pattern has been detected, which is mostly dependent on the duration of the arrhythmia, the underlying heart disease and the left atrial pressure. By transesophageal echocardiography a clear correlation has been established between the left atrial appendage slow flow and the occurrence of thromboembolic events; however, while waiting data from large studies, stratification of patients according to thromboembolic risk and decisions about anticoagulant prophylaxis should be performed by clinical information and transthoracic echocardiographic findings. No clinical or echocardiographic parameter has been found to be predictive of the thromboembolic events after cardioversion; in this setting the exclusion of atrial or atrial appendage thrombi by transesophageal echocardiography does not rule out the need for anticoagulation in patients with atrial fibrillation undergoing electrical or pharmacological cardioversion.


Percutaneous central venous catheters versus peripheral cannulae for delivery of parenteral nutrition in neonates.
Parenteral nutrition for neonates may be delivered via a short peripheral cannula or a central venous catheter. The latter may either be inserted via the umbilicus or percutaneously. Because of the complications associated with umbilical venous catheter use, many neonatal units prefer to use percutaneously inserted catheters following the initial stabilisation period. The method of parenteral nutrition delivery may affect nutrient input and consequently growth and development. Although potentially more difficult to place, percutaneous central venous catheters may be more stable than peripheral cannulae, and need less frequent replacement. These delivery methods may also be associated with different risks of adverse events, including acquired systemic infection and extravasation injury. To determine the effect of infusion via a percutaneous central venous catheter versus a peripheral cannula on nutrient input, growth and development, and complications including systemic infection, or extravasation injuries in newborn infants who require parenteral nutrition. The standard search strategy of the Cochrane Neonatal Review Group was used. This included searches of the Cochrane Central Register of Controlled Trials (CENTRAL, The Cochrane Library, Issue 1, 2007), MEDLINE (1966 - February 2007), EMBASE (1980 - February 2007), conference proceedings, and previous reviews. Randomised controlled trials that compared the effect of delivering parenteral nutrition via a percutaneous central venous catheter versus a peripheral cannulae in neonates. Data were extracted the data using the standard methods of the Cochrane Neonatal Review Group, with separate evaluation of trial quality and data extraction by each author, and synthesis of data using relative risk, risk difference and mean difference. Four trials eligible for inclusion were found. These trials recruited a total of 368 infants and reported a number of different outcomes. One study showed that the use of a percutaneous central venous catheter was associated with a decreased risk of cumulative nutritional deficit during the trial period: Mean difference in the percentage of the prescribed nutritional intake actually received: -7.1% (95% confidence interval -11.02, -3.2). In another trial, infants in the percutaneous central venous catheter group needed significantly fewer catheters/cannulae per infant during the trial period: Mean difference in the number of catheters/cannulae per infant: -3.2 (95% confidence interval -5.13, -1.27). Meta-analysis of data from all four trials did not find any evidence of an effect on the incidence of systemic infection: Typical relative risk: 0.94 (95% confidence interval 0.70, 1.25); typical risk difference: -0.02 (95% confidence interval -0.12, 0.08). Data from one small study suggest that the use of a percutaneous central venous catheter to deliver parenteral nutrition in newborn infants improves nutrient input. The significance of this in relation to long-term growth and developmental outcomes is unclear. Another study suggested that the use of a percutaneous central venous catheter rather than a peripheral cannula decreases the number of catheters/cannulae needed to deliver the nutrition. No evidence was found to suggest that percutaneous central venous catheter use increased the risk of adverse events, particularly systemic infection.


[X-ray diagnosis of primary sclerosing cholangitis].
Primary sclerosing cholangitis (PSC) should be regarded as a disease of the bile tracts which is difficult to diagnose rather than a rare disease. Combined radiodiagnostic investigation in the preoperative period is of great importance. Direct methods of an induced contrast study of the biliferous system (transcutaneous transhepatic cholangiography, endoscopic retrograde pancreatocholangiography) play a major role in the diagnosis of PSC. The authors present 11 cases, describing in detail x-ray semiotics of various sites of PSC and its differential diagnosis with similar diseases.


Cytomegalovirus mismatch still negatively affects patient and graft survival in the era of routine prophylactic and preemptive therapy: A paired kidney analysis.
The impact of cytomegalovirus (CMV) serostatus on kidney transplant outcomes in an era when CMV prophylactic and preemptive strategies are used routinely is not clearly established. Using United Network for Organ Sharing/Organ Procurement and Transplantation Network data, recipients with first deceased donor kidney transplant (≥18 years, 2010-2015) were stratified into 4 groups in the main cohort: CMV-seronegative donor (D-)/CMV-seronegative recipient (R-), CMV-seropositive donor (D+)/R-, D+/CMV-seropositive recipient (R+), and D-/R+. In a paired kidney cohort, we identified 2899 pairs of D- kidney transplant with discordance of recipient serostatus (D-/R- vs D-/R+) and 4567 pairs of D+ kidney transplant with discordance of recipient serostatus (D+/R- vs D+/R+). In the main cohort, D+/R- was associated with a higher risk of graft failure (hazard ratio [HR] = 1.17, P = .01), all-cause mortality (HR = 1.18, P < .001), and infection-related mortality (HR = 1.38, P = .03) compared with D-/R-. In the paired kidney analysis, D+/R- was an independent risk factor for all-cause mortality (HR = 1.21, P = .003) and infection-related mortality (HR = 1.47, P = .04) compared with D+/R+. No difference in graft loss between D+/R- and D+/R+. CMV mismatch is still an independent risk factor for graft loss and patient mortality. The negative impact of D+/R- serostatus on mortality persists after fully matching for donor factors.


Training in general surgery in The Netherlands.
Training in surgery has been for ages a matter of master-fellow relation between a qualified surgeon and a young physician who wishes to become a surgeon. Nowadays, however it has become a well-structured system of selection, education, training and registration, all under the responsibility of the Association of Surgeons of The Netherlands.


Inhibition of autophagy after perforator flap surgery increases flap survival and angiogenesis.
The survival ratio of multiterritory perforator flap is variable. Therefore, surviving mechanisms are increasingly explored to identify novel therapeutics. The condition of the choke zone is essential for perforator flap survival. In this study, we investigated autophagy in the choke zone after flap surgery. The flap model involved a perforator flap with three territories that was located on the right dorsal side of a rat. A total of 36 rats were divided into six groups, including the control, 0 d postoperative (PO), 1, 3, 5, and 7 d PO groups. In addition, 72 rats were divided into three groups, including a control group, a 3-methyladenine (3-MA) group, and a rapamycin group. Skin tissue of rats was used for measuring autophagy proteins, vascular endothelial growth factor (VEGF) expression, and histological examination. On day 7 after surgery, the survival ratio of each flap was determined. The expression of autophagy and VEGF in the second choke zone (choke II) was increased after flap surgery. Among the three groups, the survival ratio of flaps in the 3-MA group was the highest. Furthermore, the angiogenesis level in the 3-MA group in choke II was the highest among the three groups. Autophagy was initiated by surgery in choke II, and VEGF expression in choke II was increased after flap surgery. Inhibiting autophagy after perforator flap surgery is beneficial for flap survival and for promoting angiogenesis in choke II.


[Fine particulate matter and nonalcoholic fatty liver disease].
Fine particulate matter is defined as the particulate matter with an aerodynamic diameter of < 2.5 μm, i.e., PM2.5. Its surface absorbs large amounts of toxic and hazardous substances, which can deposit in pulmonary alveoli through respiration and reach other organs through pulmonary ventilation. Many studies have confirmed that PM2.5 is closely associated with pulmonary and cardiovascular diseases. Nonalcoholic fatty liver disease(NAFLD)has similar risk factors as these diseases, as well as obesity, hyperlipidemia, and type 2 diabetes, and it is considered a part of metabolic syndrome. In this view, many studies focus on the possible association between PM2.5 and NAFLD in recent years, including epidemiological study and experimental study, so as to investigate possible pathogenic mechanisms. With reference to the research advances in PM2.5 and NAFLD, this article reviews the association between PM2.5 and NAFLD from the aspects of lipid deposition, oxidative stress, and insulin resistance.


Cancers of the Esophagus and Stomach: Potential Mechanisms Behind the Beneficial Influence of Physical Activity.
To compare findings from several recent meta-analyses showing a reduced risk of gastric and esophageal cancers in physically active individuals, to assess the magnitude of this benefit, and to seek information on potential underlying mechanisms. A comprehensive search of Ovid/Medline from 1996 to February 2016, using the terms physical activity or exercise or training and esophageal or gastric cancer, and supplementing the articles identified by material from references lists and personal files. Moderate-to-vigorous physical activity is associated with a 20% to 30% reduction in the risk of gastroesophageal adenocarcinomas, with a significant dose/response relationship. Benefit is greater in women than in men, and greater for noncardia than for cardia or esophageal tumors. Mechanisms could include a reduction of visceral fat (with a lesser production of cancer promoting hormones and reduced gastroesophageal reflux) and/or a lesser likelihood of smoking and excessive alcohol consumption. Physical activity does not protect against Helicobacter pylori infections or gastric ulceration, but mechanisms related to the impact of exercise on immune function, antioxidant mechanisms, and gastroesophageal reflux remain to be explored. Regular, moderate-to-vigorous physical activity is associated with a clinically significant reduction in the risk of gastroesophageal adenocarcinomas, but mechanisms are as yet unclear, and a causal relationship remains to be proven.


Starch biosynthesis in cereal endosperm.
Stored starch generally consists of two d-glucose homopolymers, the linear polymer amylose and a highly branched glucan amylopectin that connects linear chains. Amylopectin structurally contributes to the crystalline organization of the starch granule in cereals. In the endosperm, amylopectin biosynthesis requires the proper execution of a coordinated series of enzymatic reactions involving ADP glucose pyrophosphorylase (AGPase), soluble starch synthase (SS), starch branching enzyme (BE), and starch debranching enzyme (DBE), whereas amylose is synthesized by AGPase and granule-bound starch synthase (GBSS). It is highly possible that plastidial starch phosphorylase (Pho1) plays an important role in the formation of primers for starch biosynthesis in the endosperm. Recent advances in our understanding of the functions of individual enzyme isoforms have provided new insights into how linear polymer chains and branch linkages are synthesized in cereals. In particular, genetic analyses of a suite of mutants have formed the basis of a new model outlining the role of various enzyme isoforms in cereal starch production. In our current review, we summarize the recent research findings related to starch biosynthesis in cereal endosperm, with a particular focus on rice.


Knockdown of Long Noncoding RNA HOXA-AS2 Suppresses Chemoresistance of Acute Myeloid Leukemia via the miR-520c-3p/S100A4 Axis.
Among different molecular candidates, there is growing data to support that long noncoding RNAs (lncRNAs) play a significant role in acute myeloid leukemia (AML). HOXA-AS2 is significantly overexpressed in a variety of tumors and associated with anti-cancer drug resistance, however, little is known regarding the expression and function of HOXA-AS2 in the chemoresistance of AML. In this study, we aimed to determine the role and molecular mechanism of HOXA-AS2 in adriamycin-based chemotherapy resistance in AML cells. Quantitative real-time PCR was used to detect HOXA-AS2 expression in the BM samples and ADR cell lines, U/A and T/A cells. Furthermore, the effects of HOXA-AS2 silencing on cell proliferation and apoptosis were assessed in vitro by CCK8 and flow cytometry, and on tumor growth in vivo. Furthermore, bioinformatics online programs predicted and luciferase reporter assay were used to validate the association of HOXA-AS2 and miR-520c-3p in AML. In this study, we showed that HOXA-AS2 is significantly upregulated in BM samples from AML patients after treatment with adriamycin-based chemotherapy and in U/A and T/A cells. Knockdown of HOXA-AS2 inhibited ADR cell proliferation in vitro and in vivo and promoted apoptosis. Bioinformatics online programs predicted that HOXA-AS2 sponge miR-520c-3p at 3'-UTR with complementary binding sites, which was validated using luciferase reporter assay and anti-Ago2 RIP assay. HOXA-AS2 could negatively regulate the expression of miR-520c-3p in ADR cells. S100A4 was predicted as a downstream target of miR-520c-3p, which was confirmed by luciferase reporter assay. Our results suggest that HOXA-AS2 plays an important role in the resistance of AML cells to adriamycin. Thus, HOXA-AS2 may represent a therapeutic target for overcoming resistance to adriamycin-based chemotherapy in AML.


Pulmonary hypoplasia and persistent pulmonary hypertension: favorable clinical response to high-frequency jet ventilation.
Pulmonary hypoplasia in the neonate is frequently fatal. This article reports two neonates with a clinical diagnosis of pulmonary hypoplasia complicated by persistent pulmonary hypertension (PPHN). Though both infants were progressively worsening while receiving conventional mechanical ventilatory support, they survived when treated with high-frequency jet ventilation.


A fluorometric assay for oxytetracycline based on the use of its europium(III) complex and aptamer-modified silver nanoparticles.
An ultrasensitive assay is described for the determination of oxytetracycline (OTC) at nanomolar levels. The method is using silver nanoparticles (AgNPs) that were first modified with OTC-binding aptamer and then exposed to the OTC-Eu(III) complex. The pink fluorescence of the OTC-Eu(III) complex on the AgNPs is almost completely quenched. On addition of OTC, it will compete with the OTC-Eu(III) complex for binding to the aptamer on the AgNPs. The OTC-Eu(III) complex is released and becomes strongly fluorescent, with excitation/emission peaks at 385/620 nm. The resulting assay was validated in terms of linearity and linear range, sensitivity, selectivity, detection limit and accuracy. Under optimum conditions, response is linear in the 10 to 500 nM OTC concentration range, and the limit of detection is 1.9 nM. The method was applied to the determination of OTC in spiked milk and tablets samples, and it gave satisfactory results. Graphical abstract Schematic presentation of the assay. In the presence of oxytetracycline (OTC), it will compete with the OTC-europium(III) complex for binding to the silver nanoparticles (AgNPs)-aptamer conjugate. The OTC-Eu(III) complex is released and strong pink fluorescence is observed.


Crystal structure of a pol alpha family replication DNA polymerase from bacteriophage RB69.
The 2.8 A resolution crystal structure of the bacteriophage RB69 gp43, a member of the eukaryotic pol alpha family of replicative DNA polymerases, shares some similarities with other polymerases but shows many differences. Although its palm domain has the same topology as other polymerases, except rat DNA polymerase beta, one of the three carboxylates required for nucleotidyl transfer is located on a different beta strand. The structures of the fingers and thumb domains are unrelated to all other known polymerase structures. The editing 3'-5' exonuclease domain of gp43 is homologous to that of E. coli DNA polymerase I but lies on the opposite side of the polymerase active site. An extended structure-based alignment of eukaryotic DNA polymerase sequences provides structural insights that should be applicable to most eukaryotic DNA polymerases.


A population dynamic approach to the cost-effectiveness analysis of mass anthelmintic treatment: effects of treatment frequency on Ascaris infection.
This study examines the cost-effectiveness of different frequencies of mass chemotherapy in reducing helminth infection and morbidity at the community level. The cost-effectiveness framework utilities a dynamic model of helminth transmission which permits the effectiveness of treatment to be expressed as some long-term reduction in infection and disease. Cost analysis is incorporated into this framework using an itemized cost menu and cost data from an actual control programme. The cost-effectiveness of mass treatment is investigated using epidemiological measures considered appropriate to Ascaris lumbricoides infection in both a high and low transmission area. The optimal strategy for control is shown to be dependent on the control objectives, the rate of transmission, and existing budget constraints. The analysis suggests that if the aim of control is to reduce morbidity due to ascariasis and mass treatment is the approach selected, then it is more cost-effective to intervene in a high transmission area than in a low transmission area, and that relatively long intervals between treatments offer the most cost-effective strategy.


Transforming and carcinogenic potential of cadmium chloride in BALB/c-3T3 cells.
A large number of workers are potentially exposed to cadmium during mining and processing. Therefore, there is a concern regarding the potential carcinogenic hazards of cadmium to exposed workers. Studies have been performed to determine if cadmium chloride (CdCl(2)) can induce morphological cell transformation, DNA from CdCl(2)-induced transformed cells can transform other mammalian cells, and the transformed cells induced by CdCl(2) can form tumors in nude mice. BALB/c-3T3 cells were treated with different concentrations of CdCl(2) for 72 h. The frequency of transformed foci from each treatment was determined after cells were cultured for 4 to 5 weeks. DNAs from five CdCl(2)-induced transformed cell lines were isolated and gene transfection assay was performed using NIH-3T3 cells. Non-transformed BALB/c-3T3 cells and cells from 10 transformed cell lines induced by CdCl(2) were injected into both axillary regions of nude mice. Mice were screened once a week for the appearance and size of tumors. CdCl(2) caused a statistically significant, concentration-related increase in the transformation frequency. DNA from all five CdCl(2)-induced transformed cell lines tested was found to induce varying degrees of transfection-mediated transformation in NIH-3T3 cells. All 10 CdCl(2)-induced transformed cell lines formed fibrosarcomas in nude mice within 39 days of inoculation. Within this time period, no tumors were found in nude mice injected with non-transformed BALB/c-3T3 cells. These results indicate that CdCl(2) is capable of inducing morphological cell transformation and that the transformed cells induced by CdCl(2) are potentially tumorigenic.


Colonic cancer in a 34-yr-old woman: should it prompt microsatellite instability studies and mismatch repair gene testing?
It remains debatable whether young patients with colorectal tumors should undergo genetic testing with the aim of identifying new hereditary nonpolyposis colorectal cancer families. We describe a case of a young woman with colon cancer with no clinical criteria of hereditary nonpolyposis colorectal cancer, whose genetic analysis showed that the tumor displayed microsatellite instability, and in whom a truncated protein in hMSH2 gene was found, which was also present in two at-risk relatives.


Immunoelectronmicroscopic localization of extracellular matrix components produced by bovine corneal endothelial cells in vitro.
Bovine corneal endothelial cells deposit an extracellular matrix in short-term cultures, which contains various morphologically distinct structures when analysed by electron microscopy after negative staining. Amongst these were long-spacing fibers with a 150 nm periodicity, which appeared also to be assembled into more complex hexagonal lattices. Another structure was fine filaments, 10-40 nm in diameter, which occasionally exhibited 67 nm periodic cross-striation. Non-striated 10-20 nm filaments sometimes formed radially oriented bundles arranged in networks and fuzzy granular material was associated with the filaments in the bundles. Often, these bundles extended into solitary filaments, 10-20 nm in diameter, with a smooth surface. In addition, amorphous patches were seen, which contained dense aggregates of fibrillar and granular material. In longer-term cultures, some of the structures coalesced to form large fibrillar bundles. By using specific antibodies to various extracellular matrix components and immunolabeling with gold some of these structures could be identified as to their protein composition. Whereas fibronectin antibodies labeled a variety of structures--fine filaments with granular materials, radially oriented bundles, patchy amorphous aggregates and small granular material scattered throughout the background--type III collagen antibody predominantly labeled filaments with periodic banding (10-40 nm in diameter). A small amount of type III specific labeling was also observed over the networks of radially oriented fibrils and fine filaments associated with granular material. Type IV collagen and laminin antibodies localized in areas of the patchy amorphous aggregates. Type VI collagen antibodies, on the other hand, labeled fine filaments and the gold particles showed a pattern of 100 nm periodicity. Many of the fine 10-20 nm filaments exhibited a tubular appearance on cross-section, but they were not reactive with any of the antibodies used. Also negative were the long-spacing fibers and assemblies--including hexagonal lattices--containing this structural element.


A case of a patient with granulocyte-colony stimulating factor-producing pancreatic cancer who responded to nab-paclitaxel plus gemcitabine.
A 67-year-old male patient presented with an irregular mass involving the pancreatic body and tail with multiple liver/lymph node metastases. A biopsy indicated the presence of a poorly differentiated adenocarcinoma. Fever and increased white blood cell count, C-reactive protein levels, and granulocyte-colony stimulating factor (G-CSF) levels led to the diagnose of G-CSF-producing pancreatic cancer. The patient did not respond to FOLFIRINOX therapy (leucovorin, fluorouracil, irinotecan, and oxaliplatin), but nab-paclitaxel plus gemcitabine treatment was effective, resulting in tumor shrinkage and reduced G-CSF levels. After the fifth course of this therapy, exacerbation was noted, and the patient died of primary cancer 6 months after initiating the therapy. Here we report the case of this patient with G-CSF-producing pancreatic cancer who responded to chemotherapy.


High-efficient nonviral transfection of the rat thyroid cell line FRTL-5.
FRTL-5 cells are used in many laboratories as an in vitro system of thyroid follicular cells since they share many properties of human thyrocytes. However, the use of FRTL-5 cells for experimental modifications is limited by low transfection efficiencies of lipid-based transfections and the need for cumbersome viral transduction protocols. A new technology - nucleofection - has become available for cell lines that are difficult to transfect. Here, we report the application and optimization of this method in FRTL-5 cells. Using the green fluorescent protein (GFP) as a reporter gene, FRTL-5 cells were easily transfectable with efficiencies over 60%. In addition, the simultaneous transfer of siRNA against GFP was feasible and allowed suppression of GFP over at least 4 days. Furthermore nucleofection was successful for establishing stable FRTL-5 cell clones. In conclusion, this optimized fast and efficient nucleofection protocol offers new properties for the experimental use of FRTL-5 cells.


[Evaluation of the quality of artesian water sources in the Moscow Region].
The investigation has confirmed that the maximum allowable concentrations (MAC) of elements, such as aluminum, manganese, fluorides, and iron, are exceeded in the artesian waters of the Moscow Region. The major man-made pollutants of portable underground water have been ascertained to be cadmium, nickel, and lead. The main site of contamination by these elements was found in the wells located in the center of the region. Hygienic assessment has shown that the Moscow Region's artesian water in 89% of cases of all tested wells fails to conform to the established standard amount of excess MAC multiplicity scores for class 1 and 2 hazard elements standardized by the sanitary toxicological indicator. According to the particular excess MAC multiplicity rating score (sigma(beta gamma)), the contamination of drinking underground water by Li, Sr, Cd, Pb, Ni, Al, Mn, Fe, and F- is detectable in the range from low to moderate levels. There is need for pre-purification of underground water for its use for drinking purposes.


Complementary Methods of Processing diS-C3(3) Fluorescence Spectra Used for Monitoring the Plasma Membrane Potential of Yeast: Their Pros and Cons.
Carbocyanine dye diS-C3(3) was repeatedly employed in monitoring the plasma membrane potential of yeast and other living cells. Four methods of measuring and evaluating probe fluorescence signal were used in different studies, based on following fluorescence parameters: fluorescence intensity emitted within a certain spectral interval, F(580)/F(560) fluorescence emission ratio, wavelength of emission spectrum maximum, and the ratio of respective fluorescence intensities corresponding to the diS-C3(3) bound to cytosolic macromolecules and remaining dissolved in the aqueous cell medium (i.e., unbound, or free). Here we show that data corresponding to the three latter spectral assessments of diS-C3(3) accumulation in cells is mutually convertible, which means that their alternative use cannot lead to ambiguities in the interpretation of the results of biological experiments. On the other hand, experiments based on the effortless measurements of fluorescence intensities should be interpreted cautiously because controversial results can be obtained, depending on the particular choice of cell-to-dye concentration ratio and emission wavelength.


Bioantioxidants: the systems reliability standpoint.
The antioxidant power of the so-called antioxidants is negligible because their rate constants and concentrations are too small to compete with the specialized defense enzymes, like superoxide dismutase (SOD), for the reactive oxygen species. In this short review, we present a number of experimental data of our group, along with the relevant literature data, to show that in-vivo antioxidants increase the systems reliability in other tacks. For example, butylated hydroxytoluene can prevent production of O2*- in mitochondria, whereas flavonoids can induce expression of antioxidant enzymes, SOD and catalase. We suggest that the timely introduction of antioxidants can provide the beneficial physiological effects through the prophylactic reliability maintenance against reactive forms of oxygen via the hormonal system.


Cardiorespiratory fitness is associated with inflammation and physical activity in HIV+ adults.
Our objective was to examine the effect of a lifestyle diet and exercise intervention on cardiorespiratory fitness (CRF) and to examine predictors of change in CRF. People living with HIV (PLHIV) are at increased risk for cardiovascular disease. CRF is a better predictor of cardiovascular disease-related mortality than established risk factors yet very little is known about CRF in PLHIV. One-hundred and seven virally suppressed PLHIV were randomized to a group-based intervention to improve lifestyle behaviors or a control condition. All PLHIV maximal cardiorespiratory stress test to determine VO2 peak, VO2 at anaerobic threshold, and ventilatory efficiency/VCO2, at baseline and 6 months later. Participants wore an accelerometer to measure physical activity, completed waist-hip circumference measures, and had a fasting lipid profile, IL-6, and high sensitivity C-reactive protein analyzed. Generalized estimating equations were used to examine the effect of the intervention on CRF and predictors of change in CRF. Participants were approximately 53 years old, 65% male (n = 70), and 86% African-American (n = 93). There was no effect of the intervention on markers of CRF over time (P > 0.05). After controlling for age, sex, waist-hip-ratio, the inflammatory biomarker IL-6 was inversely associated with a decline in both VO2 peak (P = 0.03) and VO2 at anaerobic threshold (P = 0.03). In addition, participants who walked an additional 10 000 steps per day had a 2.69 ml/kg per min higher VO2 peak (P = 0.02). Despite HIV viral suppression, PLHIV had remarkably poor CRF and inflammation was associated with a clinically adverse CRF profile. However, increased physical activity was associated with improved CRF.


Regulation of microtubule-associated proteins, protein kinases and protein phosphatases during differentiation of SY5Y cells.
Regulation of expression and function of microtubule-associated proteins (MAPs) is critical for neurons to maintain normal cytoskeletal architecture and functions. We have shown previously that in differentiated human neuroblastoma SY5Y cells, the expression of tau, a major neuronal MAP, is dramatically increased, and tau phosphorylation is differentially regulated. In the present study, we investigated the expression, the subcellular distribution and the microtubule-binding activities of several MAPs in SY5Y cells upon differentiation. We also studied the activities of protein kinases and phosphatases that are involved in regulation of tau phosphorylation during cell differentiation. We found that the expression of MAP1b in addition to tau was upregulated upon differentiation. Tau, MAP1a, MAP1b and MAP2 had distinct immunocytochemical staining patterns in differentiated SY5Y cells, suggesting differential biological functions. The microtubule-binding activity of tau increased after cell differentiation, whereas the activities of MAP1a and MAP2 decreased. Upon differentiation, the phosphorylation of tau at Ser198/Ser199/Ser202 and Ser396/Ser404 was increased, but that at Ser262/Ser356 was decreased. These changes in tau phosphorylation were accompanied by an upregulation of activities of several protein kinases (cdk5, MAPK, PKC and CK-1) as well as protein phosphatases PP-1 and PP-2A. These results suggest that the expression, post-translational modifications and biological activities of various MAPs are differentially regulated to meet the biological needs during cell differentiation.


Treatment options in imatinib-resistant chronic myelogenous leukemia.
To discuss new therapeutic options available in the treatment of chronic myelogenous leukemia (CML) in patients who failed or were intolerant to imatinib therapy. Literature was accessed via MEDLINE (1966-May 2007), EMBASE (1991-3rd quarter 2007), the Proceedings of the American Society of Hematology (2000-2006), and the Proceedings of the American Society of Clinical Oncology (2000-2007). Search terms included imatinib, dasatinib, nilotinib, and chronic myelogenous leukemia. Meeting abstracts and studies that reported preclinical and Phase 1, 2, and 3 trials published in English are included. Imatinib is the standard of care for CML; however, some patients develop resistance or are intolerant to the drug. Phase 1 and 2 clinical data for the more potent tyrosine kinase inhibitors, dasatinib and nilotinib, are promising. Hematologic and cytogenetic responses are reported with both. There does not appear to be cross-resistance between the drugs, although neither is effective against all mutations of the hallmark molecular marker, the Philadelphia chromosome. Novel agents are also being examined for the treatment of patients with CML, including aurora kinase and farnesyl transferase inhibitors, as well as combination therapies. Dasatinib and nilotinib are second-line options for patients who have CML and are resistant or intolerant to imatinib. Toxicity profiles between agents may differ. Clinical trials with these drugs and others are ongoing.


Cooperative interactions among intestinal GATA factors in activating the rat liver fatty acid binding protein gene.
GATA-4, GATA-5, and GATA-6 are endodermal zinc-finger transcription factors that activate numerous enterocytic genes. GATA-4 and GATA-6 but not GATA-5 are present in adult murine small intestinal enterocytes, and we now report the simultaneous presence of all three GATA factors in murine small intestinal enterocytes before weaning age. An immunohistochemical survey detected enterocytic GATA-4 and GATA-6 at birth and 1 wk of age and GATA-5 at 1 wk but not birth. Interactions among GATA factors were explored utilizing a transgene constructed from the proximal promoter of the rat liver fatty acid binding protein gene (Fabp1). GATA-4 and GATA-5 but not GATA-6 activate the Fabp1 transgene through a cognate binding site at -128. A dose-response assay revealed a maximum in transgene activation by both factors, where additional factor did not further increase transgene activity. However, at saturated levels of GATA-4, additional transgene activation was achieved by adding GATA-5 expression construct, and vice versa. Similar cooperativity occurred with GATA-5 and GATA-6. Identical interactions were observed with a target transgene consisting of a single GATA site upstream of a minimal promoter. Furthermore, GATA-4 and GATA-5 or GATA-5 and GATA-6 bound to each other in solution. These results are consistent with tethering of one GATA factor to the Fabp1 promoter through interaction with a second GATA factor to produce increased target gene activation. Cooperative target gene activation was specific to an intestinal cell line and may represent a mechanism by which genes are activated in the small intestinal epithelium during the period before weaning.


[Bisphosphonates in the prevention of vertebral and non-vertebral fractures in postmenopausal women with osteoporosis].
Bisphosphonates (risedronate and alendronate) inhibit bone resorption, increase bone mass and reduce the risk of vertebral and nonvertebral fractures in postmenopausal women as has been shown in a number of clinical trials. The concept of bone quality has recently been actualized with evidence that antiresorptive drugs reduce the occurrence of fractures to a greater extent than would be predicted by the amount of the increase in bone density. Concern about possible negative effect on fracture resistance with prolonged treatment with bisphosphonates has no evidence from clinical studies. Both preclinical and clinical studies, histologically and biomechanically performed, have revealed normal quality of bone formed during treatment with bisphosphonates.


Serum complement C3: a determinant of cardiometabolic risk, additive to the metabolic syndrome, in middle-aged population.
We studied whether serum complement C3 (C3) is an independent determinant of incident cardiometabolic risk (coronary heart disease [CHD], metabolic syndrome [MetS], and type 2 diabetes mellitus). A cohort of 1220 adults of a general population (age, 53 +/- 10.5 years) was evaluated prospectively at 3.3 years follow-up using Cox proportional hazard regressions. Cardiometabolic risk factors were measured. Metabolic syndrome was identified by Adult Treatment Panel III criteria modified for male abdominal obesity. The C3 levels were associated significantly and linearly with serum triglycerides, waist circumference, and C-reactive protein (CRP), and inversely with current smoking but not with the marker of insulin resistance. In regression models for incident MetS, increasing C3 quartiles strongly predicted MetS in women and in both sexes combined after adjusting for all 5 MetS components and other confounders. Circulating C3 significantly predicted in each sex incident CHD independent of age, smoking status, and presence of MetS. Even after entering CRP, C3 predicted CHD with a relative risk of 1.35 (95% confidence interval, 1.09-1.67) for 1-SD increment of C3 in the total sample. Complement C3 tended to contribute, additively to MetS, to the association with diabetes with a relative risk of 1.36 in women alone, not in men. In conclusion, elevated serum complement C3 is part of the MetS cluster and confers CHD risk, additively to MetS components and CRP, in a population in which MetS prevails. Levels contribute, additively to MetS, to the diabetes risk in women alone.


Xeroderma pigmentosum complementation group F: A rare cause of cerebellar ataxia with chorea.
The complementation group F of Xeroderma pigmentosum (XP-F) is rare in the Caucasian population, and usually devoid of neurological symptoms. We report two cases, both Caucasian, who exhibited progressive cerebellar ataxia, chorea, a mild subcortical frontal cognitive impairment, and in one case severe polyneuropathy. Brain MRI demonstrated cerebellar (2/2) and cortical (1/2) atrophy. Both patients had only mild sunburn sensitivity and no skin cancer. Mini-exome sequencing approach revealed in ERCC4, two heterozygous mutations, one of which was never described (c.580-584+1delCCAAGG, exon 3), in the first case, and an already reported homozygous mutation, in the second case. These cases emphasize that XP-F is a rare cause of recessive cerebellar ataxia and can in some cases clinically mimic Huntington's disease due to chorea and executive impairment. The association of ataxia, chorea, and sun hypersensitivity are major guidance for the diagnosis, which should not be missed, in order to prevent skin neoplastic complications.


The voices of Brazilian women breaking free from intimate partner violence.
Intimate partner violence (IPV) is a global problem with a direct effect on the health of the victims and their communities. With the intention to inform nursing care delivered to women experiencing IPV, this qualitative study was conducted, specifically to understand the meaning that IPV has for assaulted women and to identify the factors that contribute to breaking the cycle of violence. Data collection included interviews and discussion of photographs. Participants were recruited from within a shelter for women experiencing violence. Findings from five women were inductively coded into three categories: "experiencing violent acts," which describes the period in which the women lived with the aggressor; "marks of the violent acts," which portrays the consequences of IPV; and "women's support," which indicates the current supports or lack thereof for these women. Nurses and other healthcare providers will benefit from hearing the women's voices through their words and photographs and will recognize the need for holistic care that includes the physical, sexual, and emotional needs of abused women.


Effect of sodium fluoride on early stages of otosclerosis.
The present investigation evaluates the effectiveness of NaF treatment in modifying the natural course of subclinical otosclerosis, as monitored by the stapedius reflex. The study was carried out on 128 relatives of patients suffering from surgically confirmed footplate otosclerosis. The diagnosis of subclinical otosclerosis was made on the basis of presence of the on-off effect. One group of subjects was treated with NaF in doses ranging from 6 to 16 mg according to age. The treatment lasted 2 years. A second group served as a control. Changes in stapedius reflex morphology were evaluated at 1, 2, and 5 years from the onset of treatment. The investigation demonstrated that NaF has a stabilizing effect on early otosclerosis. This drug, in fact, arrests the disease process in more than 60 percent of ears at the 2-year follow-up and in more than 50 percent at 5 years. A program of secondary prevention of otosclerosis by NaF is suggested.


Diffuse muscoskeletal pain and proximal myopathy: do not forget hypovitaminosis D.
A 20-year-old vegetarian man was admitted to our hospital complaining of muscle weakness and gait disturbances of 4 years duration. For the past 5 years, he had major depression and had confined himself at home. He exhibited tenderness upon palpation of the chest, sternum and proximal muscles. Hypocalcemia, hypophosphatemia, vitamin D deficiency, increased levels of alkaline phosphatase, and intact parathyroid hormone were noted. An x-ray skeletal survey revealed generalized osteopenia, multiple vertebral and costal fractures, and a pelvis deformed into the shape of a triangle. A diagnosis of osteomalacia secondary to vitamin D deficiency from lack of exposure to sunlight and to inadequacy of the diet was made. The patient was started on a treatment with 20,000 IU of vitamin D3 once a week plus 1 g/d of calcium. Eight months later, gait disturbances have significantly improved and laboratory findings have all normalized.


Upcoming biologic agents for the treatment of rheumatic diseases.
The development of biologic agents has provided rheumatologists with a variety of new and effective treatment options. The success of early biologics, especially etanercept and infliximab for the treatment of rheumatoid arthritis, has spurred research into novel targets for the management of systemic inflammatory and autoimmune diseases. In addition, existing biologics approved for use in other diseases, such as rituximab, are now under study for the treatment of new indications. This article reviews ongoing research on the treatment of rheumatic diseases with new and existing biologic agents.


Allosteric regulation in a family of enterobacterial aspartate transcarbamylases: intramolecular transmission of regulatory signals in chimeric enzymes.
Several enterobacterial aspartate transcarbamylases (ATCases) exhibit a [2(C3):3(r2)] quaternary structure analogous to that of the Escherichia coli enzyme. Despite their conserved quaternary structures, these enzymes present substantial differences in the co-operativity of substrate binding and in their allosteric regulation by nucleotide effectors. A comparison between different enzymatic species provides an opportunity to expand our understanding of the molecular basis of allostery in ATCase. Chimeric ATCases were constructed by exchanging subdomain regions involved in quaternary structural features, such as the r1-c4 regulatory-catalytic subunit interface analyzed in this study, in order to define the involvement of this interface in the several components of allosteric regulation. The r1-c4 interface was found to constitute an essential element for the recognition and the transmission of the ATP regulatory signal in the Serratia marcescens and the Proteus vulgaris ATCases, as it does in the E. coli ATCase. Besides, the specific amino acid composition of the C-terminal region of the regulatory chain and its interactions with the amino acid residues in the 240s loop of the catalytic chain (r1-c4 interactions) were found to modulate the amplitude of the enzyme's response to ATP. The C-terminal region of the regulatory chain did not appear to participate directly in the regulation of the three native ATCases by CTP. Even when CTP acts as an activator, as in the P. vulgaris and S. marcescens ATCases, its signal follows a route distinct from that of the general activator ATP. Synergistic inhibition by CTP and UTP was found to involve the transmission of a specific UTP signal. This signal appeared different in the various ATCases, involving the C-terminal region of the regulatory chain in the E. coli and S. marcescens ATCases but not in the P. vulgaris ATCase.


DNA sequence of the promoter region of the ompC gene and the amino acid sequence of the signal peptide of pro-OmpC protein of Escherichia coli.
The osmoregulated ompC gene of Escherichia coli was cloned and the DNA sequence of a fragment encompassing the promoter region and a portion of the coding region was determined. There were no obvious homologies in the DNA sequences of the promoter regions of the ompC and ompF genes, in contrast to those of the coding regions of the two genes, both of which code for the matrix porins (major outer membrane proteins) and form passive diffusion pores. The amino acid sequence of the signal peptide of pro-OmpC protein was also deduced from the DNA sequence.


Early diagnosis and treatment of leprosy in intradomiciliary contacts in a high prevalence area: Amazon region.
The importance of dermato-neurological examination of intradomiciliary contacts is well known as an important secondary preventive measure in leprosy control, due to the fact that it allows early diagnosis and treatment. This is an intervention trial in an area of high leprosy prevalence (Manaus/Brazil) where the proportion of contacts examined is low. The aim of the study is to assess whether a simple educational session conducted among patients increases contacts examination and leads to early case detection. The intervention group had examined more contacts (p < 0.05) but, paradoxically, presented fewer new cases than the control group. The authors discuss the probable causes for this unexpected outcome, the advantages of the intervention and other related issues.


Paul Ehrlich: the Nobel Prize in physiology or medicine 1908.
We wish to commemorate Paul Ehrlich on the centennial of his being awarded the Nobel Prize in Physiology or Medicine in 1908. His studies are now considered as milestones in immunology: the morphology of leukocytes; his side-chain theory where he defined the cellular receptor for first time; and his clarification of the difference between serum therapy and chemotherapy. Ehrlich also invented the first chemotherapeutic drug: compound 606, or Salvarsan. We have used some original documents from the Royal Society of London, where Ehrlich was a fellow, and from Leipzig University, where he took a degree in medicine.


The role of a lipido-sterolic extract of Serenoa repens in the management of lower urinary tract symptoms associated with benign prostatic hyperplasia.
OBJECTIVE To examine the clinical profile of medication derived from a lipido-sterolic extract of Serenoa repens (saw palmetto) for managing lower urinary tract symptoms (LUTS) associated with benign prostatic hyperplasia (BPH). METHODS We reviewed clinical trials involving extracts of S. repens, focusing on the benefit/risk ratio in patients with BPH. RESULTS S. repens extract significantly reduces the symptoms of BPH, increases urinary flow, improves the quality of life and is well tolerated. CONCLUSION Analysis of the overall clinical database indicates that extract of S. repens may be considered a viable first-line therapy for treating LUTS.


Effects of clonidine and naloxone on the dopamine levels in the rat mediobasal hypothalamus measured by in vivo voltammetry.
We examined the effect of alpha 2-adrenergic stimulation on dopamine (DA) levels in the mediobasal hypothalamus (MBH) by means of in vivo voltammetry in the rat. Intravenous (i.v.) injection of an alpha 2-agonist, clonidine (15 micrograms/100 g b.wt.) caused a decrease of MBH DA levels (76 +/- 3% of basal levels). Pretreatment with phenoxybenzamine (500 micrograms/100 g b.wt.i.v.), an alpha-antagonist, blunted the decrease of MBH DA levels induced by clonidine. Naloxone (125 micrograms/100 g b.wt.i.v.), an opioid antagonist, blunted the decrease of MBH DA levels induced by clonidine. These results suggest that alpha 2-adrenergic mechanisms may inhibit the tuberoinfundibular dopaminergic (TIDA) system through the activation of the endogenous opioid system.


Seasonal, Locality, and Habitat Variation in Assemblages of Carrion-Associated Diptera in Gauteng Province, South Africa.
Seasonal, spatial, and habitat responses of carrion-associated Diptera assemblages can provide valuable information about the presence or absence of species and their relative abundance, and thereby enhance understanding of their responses to environmental variables and how this may have an impact on forensic investigations. Three different nature reserves (localities) within the Municipality of Tshwane, South Africa, were selected to determine whether species assemblages of carrion-feeding flies differ between seasons, localities, and habitat types. A total of 59,511 adult Diptera, identified to 35 species in eight different families, were collected using modified Redtop hanging traps, baited with liver and fish, during four seasons in three different habitat types. Species assemblages differed temporally, with season being the main factor determining species diversity and not locality or habitat. However, savanna and human-disturbed habitats supported a higher abundance and species richness than grassland habitats. Areas adjacent to the localities, such as large urban expanses in Dinokeng or agricultural holdings in Rietvlei, led to an increase in the abundance and mean species richness of carrion-associated Diptera, and in increased numbers of pest or invasive species such as Chrysomya megacephala (F.). Despite this, the overall species assemblages present in human-disturbed areas were very similar to those recorded in natural habitats.


Efficacy of 5-vinyl-1-beta-D-arabinofuranosyluracil (VaraU) against herpes simplex virus type 2 strains in cell cultures and against experimental herpes encephalitis in mice: comparison with acyclovir and foscarnet.
The sensitivity of different herpes simplex virus type 2 (HSV-2) strains to inhibition by 5-vinyl-1-beta-D-arabinofuranosyluracil (VaraU) was evaluated in comparison to 9-(2-hydroxyethoxymethyl)guanine (ACV; acyclovir) and trisodiumphosphonoformate (Na3PFA; foscarnet), using a plaque inhibition assay in primary rabbit testes (PRT) cells as well as in human embryonic lung fibroblast (HELF) cell cultures. The order of decreasing activity found was ACV much greater than VaraU greater than Na3PFA in PRT cells and ACV greater than VaraU much greater than Na3PFA in HELF cells, with 50% inhibition doses (ID50) of 1.8, 8.8, and greater than 110 microM for the three drugs in HELF cells, respectively. After 72hr of drug treatment, inhibition of HELF cell proliferation by VaraU (ID50, greater than 1000 microM) was less than that by ACV and Na3PFA, resulting in high selectivity indexes of greater than 100 against HSV-2 for VaraU and ACV. Their in vivo efficacy was assessed in a mouse encephalitis model. Using a treatment schedule of three daily intraperitoneal (ip) doses over a period of 5 days, only the survival times of mice were considerably prolonged by VaraU (150 or 300 mg/kg per day; P less than 0.05 or P less than 0.001, respectively). In contrast, ACV treatment (150 mg/kg per day) led to a nearly complete prevention of encephalitis and death (P less than 0.001). Similar therapy results with VaraU application through the drinking water were obtained using only one-sixth of the high ip dose (approximately 50 mg/kg per day) but over a prolonged period of treatment. Under similar conditions no therapeutic effect of oral Na3PFA was observed.


Foreign body aspiration.
To analyze the clinical and radiological profile of foreign body aspiration in children reaching a tertiary care center and identify areas of possible interventions for proper management of such cases. Tertiary level teaching hospital. Case records of patients suspected to have foreign body aspiration over the past four years were analyzed. Clinico-radiological features, types and location of foreign bodies were studied. Of 75 children who underwent rigid bronchoscopy, 70 had tracheo-bronchial foreign bodies. History of choking was elicited in 90 percent cases. In 30 percent cases chest radiographs were non-contributory, while the commonest finding (63 percent) was distal emphysema. Over three-fourth of the cases were below the age of 2 years. Vegetative foreign bodies, mainly peanuts, were commonly present. In many cases, referral was delayed as the diagnosis was missed initially. Foreign body aspiration remains a common unintentional childhood injury due to improper exposure of young children to otherwise innocuous looking nuts and other small objects.


Preparation and electrochromic property of covalently bonded WO3/polyvinylimidazole core-shell microspheres.
Covalently bonded WO3/polyvinylimidazole (C-WO3/PVI) core-shell microspheres in sizes of 250 nm were prepared. The microstructures of C-WO3/PVI core-shell microspheres were characterized by TEM, IR, and XRD. It is found that the chemical and thermal stabilities of C-WO3/PVI core-shell microspheres are higher than those of pure WO3 nanoparticles and noncovalently bonded WO3/polyvinylimidazole (NC-WO3/PVI) core-shell microspheres. This is attributed to the strengthened interaction of the WO3 nanoparticle core and the PVI shell resulting from the interaction of covalent bonds. The electrochromic device made by the C-WO3/PVI core-shell microspheres was studied. It is suggested that the C-WO3/PVI core-shell microspheres exhibit better electrochromic properties than pure WO3 nanoparticles or NC-WO3/PVI core-shell microspheres.


Apparent heterogeneity in the pIII-peptide fusion protein in single-phage clones isolated from peptide libraries.
Ligand homogeneity is an important issue in affinity chromatography. Using phages expressing peptides on the pIII protein, a heterogeneity in the binding of monoclonal phages was observed during affinity chromatography on supermacroporous cryogels. Fractions with different apparent binding affinities could be separated by stepwise elution. When these different fractions were re-applied, the respective differences in affinity were retained. However, when phage fractions with different apparent affinities were first amplified, an offspring was generated with again variable affinities. As the sequence of the peptide insert was the same, the heterogeneity must be ascribed to differences in avidity and although no direct evidence could be generated, we hypothesize that this is possibly due to phages displaying different numbers of the same peptide as a consequence of either proteolytic or packaging events during the amplification step in Escherichia coli.


In situ studies on the switching behavior of ultrathin poly(acrylic acid) polyelectrolyte brushes in different aqueous environments.
The pH-dependent switching of a poly(acrylic acid) (PAA) polyelectrolyte brush was investigated in situ using infrared spectroscopic ellipsometry (IRSE). The brush was synthesized by a "grafting to" procedure on silicon substrate with a native oxide layer. The overall thickness of the PAA brush in the dry state was approximately 5 nm. Reversible switching of the polymer brush was studied at titration from pH 2 to 10 and back in steps of 1 pH unit. The switching process was observed by monitoring the characteristic vibrational bands of the carboxylic groups of the PAA molecules. Decreasing of the C=O vibrational band amplitude and arising of a COO(-) vibrational band proved the chemical changes in the molecular structure of the brushes due to changes of the pH value in the aqueous solution. Due to the strong absorption of these bands in the IR region, the switching process could be monitored clearly. Switching the brush in several cycles with increasing and decreasing pH value showed a hysteresis-like behavior. For the first time, such hysteresis is observed in titration experiments of polyelectrolyte brushes. This behavior is attributed to the complex mechanisms of the ion's mobility in the brush layer which is explained with a suggested simplifying model describing the influence of ions inside the brush layer. In addition to the IRSE measurements, X-ray standing waves (XSW), in situ visible ellipsometry, and contact angle measurements have been performed and were in good agreement with the results from IRSE. Repetition of the in situ measurement cycles proved the good reversibility of the switching process which is highly important for practical applications of polymer brushes.


River temperature sensitivity to hydraulic and meteorological parameters.
A sensitivity analysis is performed to evaluate river temperature variations in response to changes in hydraulic and meteorological conditions. The effects of instream flow, river geometry, and weather factors on daily mean and daily maximum river temperatures are quantified by analytical solutions to a simplified model. The influence coefficient method is used to determine river temperature sensitivity. The sensitivity analysis presents quantitative evidence that river temperatures are more sensitive to instream flowrate, upstream inflow temperature, air temperature, humidity and solar radiation than to other parameters including wind speed and channel geometry and morphometry. It is found that the sensitivity of river temperatures to flow is as significant as that to weather. Daily maximum river temperature is more sensitive to flowrate than daily mean temperature. Adapting the concept of 'diminishing returns', a critical instream flowrate is identified, which divides high and low sensitivity of water temperatures to flowrate. The critical flowrate can be used to determine practically achievable and economically feasible flow requirements for summer river temperature control. The sensitivity results can assist in streamflow management and reservoir operation for protections of habitat and aquatic environment.


Synthesis, characterization and properties of a family of lead(II)-organic frameworks based on a multi-functional ligand 2-amino-4-sulfobenzoic acid exhibiting auxiliary ligand-dependent dehydration-rehydration behaviours.
A systematic investigation is reported of the use of the multi-functional ligand 2-amino-4-sulfobenzoate (asba(2-)) to construct lead(II)-organic frameworks in the presence and absence of N-donor auxiliary ligands phen, bipy and bix [phen = 1,10-phenanthroline, bipy = 2,2'-bipyridine and bix = 1,4-(methylene-benzene)bisimidazole]. Under ambient, aqueous conditions the assembly of asba(2-) with Pb(II) and phen or bipy leads to two iso-structural 2D double-layer frameworks, {[Pb2(asba)2(phen)2(H2O)]·2H2O}n (1) and {[Pb2(asba)2(bipy)2(H2O)]·2H2O}n (2). [Pb2(asba)2(bix)2(H2O)2]n (3) is obtained in the presence of the auxiliary ligand bix and possesses a 3D network built up from 2D Pb(II)-asba(2-)-bridged double-layer pillared by bix. A 2D (4,4) topological network [Pb(asba)(H2O)]n (4) is obtained in the absence of any second ligand or presence of some bistriazole bridging spacers. The coordination modes of the ligand asba(2-) in 1-4 are greatly dependent on the type of auxiliary ligand and the compounds exhibit auxiliary ligand-dependent dehydration-rehydration behaviours; 1 shows in situ rapid and reversible dehydration-rehydration behaviour in air, while the iso-structural compound 2 loses crystallinity in air and transforms into {[Pb2(asba)2(bipy)2(H2O)]·H2O}n (2A) verified by TGA, elemental analysis and powder X-ray diffraction analysis (PXRD). 3 also shows reversible dehydration-rehydration behaviour, but it takes around one week to rehydrate even after exposure to water vapor, while the dehydrated phase of 4 rehydrates to a new crystalline material. 1 and 3 fall within the category of the "recoverable collapsing" and "guest-induced re-formation" frameworks. The water molecules in 1 and 3 have some influence on their solid state fluorescent emission intensity.


A narrative review on effects of vitamin D on main risk factors and severity of Non-Alcoholic Fatty Liver Disease.
The global prevalence of Non-alcoholic fatty liver disease (NAFLD) is increasing rapidly. Many studies have been conducted on the treatment of NAFLD; nevertheless, there is still no approved drug treatment for this disease. Although the pathogenesis of NAFLD is not fully understood, but inflammation, insulin resistance, oxidative stress, obesity and dyslipidemia are among the main causes. Epidemiological studies have shown that hypovitaminosis D is associated with these factors causing NAFLD. In addition, rate of Vitamin D deficiency has been shown to be directly related to the severity of NAFLD. Accordingly, it is believed that vitamin D may help to treatment of NAFLD by improving the above-mentioned risk factors. The purpose of this review is to survey the recent advances in the field of Vitamin D efficacy on risk factors and the severity of NAFLD based on existing evidence, especially the clinical efficiency of vitamin D supplementation in patients with NAFLD.


Validation of 8-[123I]iodo-L-1,2,3,4-tetrahydro-7-hydroxyisoquinoline-3-carboxylic acid as an imaging agent for prostate cancer in experimental models of human prostate cancer.
Very few tracers are currently available for the detection and staging of prostate cancer with positron emission tomography and single-photon emission computed tomography. This study evaluates the potential of 8-[123I]iodo-1,2,3,4-tetrahydro-7-hydroxyisoquinoline-3-carboxylic acid [ITIC(OH)] as an imaging agent for prostate cancer in experimental models of human prostate cancer. ITIC(OH) was prepared by the IODO-GEN method, with 82+/-7% radiochemical yield and >99% radiochemical purity after high-performance liquid chromatography. Thereafter, ITIC(OH) was examined in CD-1 nu/nu mice engrafted with human PC-3 and DU-145 prostate cancer in the flank or orthotopically in the prostate. Bioevaluation involved examination of the in vivo stability and uptake characteristics of ITIC(OH) into tumors and different organs by dynamic in vivo analysis and gamma counting of organs of interest after dissection. ITIC(OH) showed good in vivo stability for biological investigations and was primary cleared through urine. In vivo, ITIC(OH) accumulated highly and specifically in tumors, reaching 13.6+/-2.1% to 16.2+/-2.5% injected dose per gram (ID/g) in heterotopic tumors compared with 14.8+/-2.6% and 17.6+/-3.4% ID/g in orthotopic tumor engrafts at 60 and 240 min postinjection, respectively. In contrast, radioactivity uptake in the blood, spleen, liver and gastrointestinal tract was moderate and decreased with time, resulting in marked tumor-to-background and excellent visualization of tumors. These results suggest that ITIC(OH) is a promising candidate as radiotracer for detecting prostate cancer and warrants further studies in patients to ascertain its potential as an imaging agent for clinical use.


Vascular endothelium does not activate CD4+ direct allorecognition in graft rejection.
Expression of MHC class II by donor-derived APCs has been shown to be important for allograft rejection. It remains controversial, however, whether nonhemopoietic cells, such as vascular endothelium, possess Ag-presenting capacity to activate alloreactive CD4(+) T lymphocytes. This issue is important in transplantation, because, unlike hemopoietic APCs, allogeneic vascular endothelium remains present for the life of the organ. In this study we report that cytokine-activated vascular endothelial cells are poor APCs for allogeneic CD4(+) T lymphocytes in vitro and in vivo despite surface expression of MHC class II. Our in vitro observations were extended to an in vivo model of allograft rejection. We have separated the allostimulatory capacity of endothelium from that of hemopoietic APCs by using bone marrow chimeras. Hearts that express MHC class II on hemopoietic APCs are acutely rejected in a mean of 7 days regardless of the expression of MHC class II on graft endothelium. Alternatively, hearts that lack MHC class II on hemopoietic APCs are acutely rejected at a significantly delayed tempo regardless of the expression of MHC class II on graft endothelium. Our data suggest that vascular endothelium does not play an important role in CD4(+) direct allorecognition and thus does not contribute to the vigor of acute rejection.


BRCA mutations cause reduction in miR-200c expression in triple negative breast cancer.
Triple negative breast cancer (TNBC) is the most aggressive and poorly understood subclass of breast cancer (BC). Over the recent years, miRNA expression studies have been providing certain detailed overview that aberrant expression of miRNAs is associated with TNBC. Although TNBC tumors are strongly connected with loss of function of BRCA genes, there is no knowledge about the effect of BRCA mutation status on miRNA expressions in TNBC cases. The aims of this study were to evaluate the expression profile of miRNAs that plays role in TNBC progression and the role of BRCA mutations in their regulation. The expression level of BC associated 13 miRNAs was analyzed in 7 BRCA mutations positive, 6 BRCA mutations negative TNBC cases and 20 non-tumoral tissues using RT-PCR. According to RT2 Profiler PCR Array Data Analysis, let-7a expression was 4.67 fold reduced in TNBCs as compared to normal tissues (P=0.031). In addition, miR-200c expression was 5.75 fold reduced in BRCA mutation positive TNBC tumors (P=0.005). Analysis revealed a negative correlation between miR-200c and VEGFA expressions (r=-468). Thus, miR-200c may be involved in invasion and metastasis in TNBC cases with BRCA mutation. In this study we provide the knowledge on the first report of association between microRNA-200c and BRCA mutations in TNBC. Further studies and evaluations are required, but this miRNA may provide novel therapeutic molecular targets for TNBC treatment and new directions for the development of anticancer drugs.


The effect of intracellular pH on the rate of hexose uptake in Chlorella.
The rate of hexose uptake by Chlorella is reduced by uncouplers such as carbonyl cyanide p-trifluoromethoxyphenyl hydrazone or dinitrophenol even before concentration equilibrium is reached. The addition of uncouplers changes the membrane potential and the intracellular pH. The membrane potential does not influence the initial velocity of net sugar uptake, whereas manipulation of the cell pH by means of dimethyloxazolidinedione or by butyric acid uncovered a dramatic influence of cell pH on the rate of hexose uptake: at pH values of 7.5--6.8 maximal rate of uptake is observed but at more acid pH a strong inhibition takes place with virtually total blockage of uptake at pH 6.1. The decrease of cell pH to 6.1 in the presence of carbonyl cyanide p-trifluoromethoxyphenyl hydrazone could therefore account for the decrease in hexose transport rate. It was shown that the intracellular pH as such determines the rate of uptake and not the pH difference between inside and outside; the transport rate did not correlate with delta pH.


[Congenital valves of the posterior urethra in puberty with primary enuresis as the only symptom].
To present a case of a 16-year-old male patient with primary enuresis refractory to conservative treatment. Radiologic and urodynamic tests revealed posterior urethral valves that were treated by transurethral fulguration. The patient was cured of both enuresis and infravesical obstruction and remains disease-free 3 years after the operation with no impact on his sexual function. Posterior urethral valves are very rarely diagnosed in adolescents and adults. Very few cases have been published in the literature. To our knowledge, the case described herein is the first case presenting with persistent primary enuresis.


Vascular risk factors as treatment target to prevent cognitive decline.
Epidemiological studies have consistently shown that vascular risk factors including hypertension, diabetes, obesity, hypercholesterolemia, smoking, and lack of physical exercise are associated with an increased risk of cognitive decline and dementia. Neuroradiological and neuropathological studies have confirmed the importance of cerebrovascular lesions in the etiology of late onset dementia. We have reviewed the literature and conclude that up until now randomized controlled clinical trials targeting individual risk factors and assessing cognitive decline or dementia as an outcome have not convincingly shown that treatment of vascular risk factors can actually prevent or postpone cognitive decline and dementia. New studies targeting several vascular risk factors at the same time and using cognitive decline or dementia as primary outcome might answer the question whether cognitive decline can really be postponed or even prevented. The design of such studies is not straightforward and long follow-up is required. In this review we discuss several pertinent methodological issues that need to be addressed to achieve an optimal design of new randomized controlled trials.


Downregulation of the renin-angiotensin system in streptozotocin-diabetic rats.
To determine if insulin has the ability to regulate components of the renin-angiotensin system, renin and angiotensinogen mRNA and plasma concentrations were determined in 4-wk streptozotocin (STZ)-diabetic rats. In another group of STZ-diabetic rats, replacement insulin therapy was given over the 4-wk period, and the above parameters were examined. In STZ-diabetic rats, there was a significant regression of white adipose tissue that was accompanied by an increase in the yield of RNA obtained. Changes in white adipose tissue were reversed by insulin replacement therapy in STZ-diabetic rats. There were no changes in brown adipose tissue weight or RNA yield in STZ-diabetic rats. Plasma renin activity (PRA) was significantly decreased in STZ-diabetic rats; however, plasma angiotensinogen concentration was not significantly affected by diabetes. PRA was restored to control levels in STZ-diabetic rats with insulin replacement. Kidney renin mRNA as well as liver, epididymal, and interscapular fat angiotensinogen mRNA were significantly decreased in STZ-diabetic rats. Renin and angiotensinogen mRNA were not significantly different from control in all tissues examined in STZ-diabetic rats with insulin replacement therapy. Results from this study suggest a downregulation of the renin-angiotensin system in 4-wk STZ-diabetic rats at the level of mRNA expression that is restored by replacement therapy with insulin; therefore, insulin may directly or indirectly regulate the renin-angiotensin system.


Synthesis and binding properties of oligonucleotides covalently linked to an acridine derivative: new study of the influence of the dye attachment site.
2-Methoxy-6-chloro-9-aminoacridine has been coupled via a polymethylene linker to various positions of an oligonucleotide chain: the 3'-position, using a new universal support, the 5'-position, and both 5'- and 3'-positions via a phosphate. The intercalating agent was also linked to the oligonucleotide chain via an internucleotide phosphorothiolate. The mixture of diastereoisomers was obtained as well as each pure Rp and Sp isomer. Finally, the acridine moiety was introduced to the 5-position of the deoxyuridine. The binding properties of these oligonucleotide-acridine conjugates with their DNA counterparts have been studied by absorption spectroscopy.


Preferential expansion of human virus-specific multifunctional central memory T cells by partial targeting of the IL-2 receptor signaling pathway: the key role of CD4+ T cells.
Effector memory T cells are effective in controlling acute infections, but central memory T cells play a key role in long-lasting protection against viruses and tumors. In vivo/in vitro challenge by Ag commonly supports the generation of effector memory T cells with limited longevity. To our knowledge, this study demonstrates for the first time in the human system and under rechallenge conditions that targeting IL-2R by partial mammalian target of rapamycin inhibition or blocking IL-2Rα enriches human CD4(+)/CD8(+) central memory T cells within the virus-specific T cell product associated with enhanced functionality (i.e., multicytokine secretors, including IL-2; enhanced CD137 and CD107a expression on CD8(+) and CD4(+) T cells, respectively; and killing infected target cells). Remarkably, the effects on CD8(+) T cells are mainly mediated via the enhancement of CD4(+) T cell function. The data reveal new insights into the role of CD4(+) T cell support for the quality of CD8(+) T cell memory, even under rechallenge conditions. Moreover, our method offers a new approach to improve the long-lasting efficacy of adoptive T cell therapy in patients.


Innovations in generation and analysis of 2D [(13)C,(1)H] COSY NMR spectra for metabolic flux analysis purposes.
2D [(13)C,(1)H] COSY NMR is used by the metabolic engineering community for determining (13)C-(13)C connectivities in intracellular compounds that contain information regarding the steady-state fluxes in cellular metabolism. This paper proposes innovations in the generation and analysis of these specific NMR spectra. These include a computer tool that allows accurate determination of the relative peak areas and their complete covariance matrices even in very complex spectra. Additionally, a method is introduced for correcting the results for isotopic non-steady-state conditions. The proposed methods were applied to measured 2D [(13)C,(1)H] COSY NMR spectra. Peak intensities in a one-dimensional section of the spectrum are frequently not representative for relative peak volumes in the two-dimensional spectrum. It is shown that for some spectra a significant amount of additional information can be gained from long-range (13)C-(13)C scalar couplings in 2D [(13)C,(1)H] COSY NMR spectra. Finally, the NMR resolution enhancement by dissolving amino acid derivatives in a nonpolar solvent is demonstrated.


Reversibility of the adverse effects of 1-bromopropane exposure in rats.
Previous experiments indicated that 1-bromopropane (1-BP), an alternative to chloroflurocarbons, is neurotoxic and inhibits spermiation in the testis. Here we investigated the reversibility of the toxic effects of 1-BP in rats. Male Wistar rats were divided into three equal groups of 24 each and exposed by inhalation to 0, 400 or 1000 ppm of 1-BP for 6 weeks (8 hrs/day, 7 days/week). Eight rats from each group were sacrificed at the end of 6 weeks exposure, and at 4 and 14 weeks after the end of exposure, to assess the recovery processes. We studied sperm count, motility, morphology and testicular histopathology, as well as blood pressure, skin temperature and hindlimb muscle strength. At the end of 6 weeks of exposure to 1000 ppm (0 week recovery), testicular weight, epididymal weight, sperm count and motility were low, morphologically abnormal sperm were increased and spermatogenic cells showed diffuse degeneration. These changes did not show full recovery at 14 weeks recovery, with the exception of the prostate and seminal vesicular weights, which recovered back to control values. At 400 ppm, increased retained spermatids at 0 week recovery returned to normal at 4 weeks recovery. Exposure to 1000 ppm produced sustained reduction of hindlimb muscle strength at 14 weeks recovery, whereas normalization of the skin temperature and blood pressure was noted after transient changes. Our study showed that the effect of 1-BP on spermatogenesis is dose-dependent; low exposure inhibited spermiation and hormone-dependent organ weight reduction and these changes were transient, while a higher dose of 1000 ppm 1-BP caused persistent depletion of spermatogenic cells.


Predictive modeling of non-viral gene transfer.
In non-viral gene delivery, the variance of transgenic expression stems from the low number of plasmids successfully transferred. Here, we experimentally determine Lipofectamine- and PEI-mediated exogenous gene expression distributions from single cell time-lapse analysis. Broad Poisson-like distributions of steady state expression are observed for both transfection agents, when used with synchronized cell lines. At the same time, co-transfection analysis with YFP- and CFP-coding plasmids shows that multiple plasmids are simultaneously expressed, suggesting that plasmids are delivered in correlated units (complexes). We present a mathematical model of transfection, where a stochastic, two-step process is assumed, with the first being the low-probability entry step of complexes into the nucleus, followed by the subsequent release and activation of a small number of plasmids from a delivered complex. This conceptually simple model consistently predicts the observed fraction of transfected cells, the cotransfection ratio and the expression level distribution. It yields the number of efficient plasmids per complex and elucidates the origin of the associated noise, consequently providing a platform for evaluating and improving non-viral vectors.