Cancer chemoprevention by supplemental carotenoids and synergism with retinol in mastodynia treatment.
Cancer chemoprevention with beta-carotene (BC), canthaxanthin (CX) and retinol-BC is reported with respect to skin, breast, gastric, colon carcinogeneses induced by benzo(a)pyrene (BP) with or without ultra violet radiation (UV-A, UV-B), dimethylbenzathracene (DMBA) +/- UVB, P-UVA, N-methyl-N'-N-nitro-nitrosoguanidine (MNNG), dimethylhydrazine (DMH), and with respect to transplanted tumours. When animals were loaded with carotenoid supplementation one month before the carcinogenic induction (continued throughout the experiment), cancer prevention was observed up to 60-100%. The absence of provitamin A-activity in CX shows the carotenoid antioxidant property. Fifteen patients given BC + CX to prevent recurrences after radical removal of the primary neoplasia in organs like lung, breast, colon, urinary bladder, head and neck were studied in 1980-89. A longer than expected disease-free interval was preliminarily found. Supplementation of BC +/- retinol was also reported to prevent and treat oral leucoplakia. Supplementation and intermittent retinol administration was also tested in benign cyclical mastalgia with clear cut side effect free therapeutic results.


Effects of droughts on health: Diagnosis, repercussion, and adaptation in vulnerable regions under climate change. Challenges for future research.
There is little doubt about the effects of drought events on human health in the present climate. Projections of climate change indicate an increase in the occurrence and severity of droughts in the 21st century in a number of regions, thus it is likely that these types of hydrological extremes could have more of an impact if appropriate adaptation measures are not taken. The majority of studies on the effects of drought are focused on meteorological, agricultural, or hydrological contexts, but there are rather fewer assessments of the impacts of droughts on health. In particular, there have been hardly any attempts to compare different drought indices in order to identify and quantify the impacts of drought on health systems. In addition, rather better knowledge is needed on the mechanisms of vulnerability involved. In this paper, we attempt to describe the complexity of drought phenomena and the difficulty involved in quantifying the health risks linked to their occurrence. From an international perspective, we provide a brief review of the harmful effects of droughts on health in the context of climate change, as well as the vulnerability factors related to droughts. We make an assessment of aspects that have not yet been investigated, or which require further attention to be devoted to this topic. The principal aim of this paper is therefore to draw attention to the need to consider closely the relationship between drought indices and human health, in order to achieve a more fundamental understanding, and to propose specific courses or lines of action for future years, which could eventually be of use to healthcare providers and services.


Too Many Shades of Grey: Photometrically and Spectrally Mismatched Targets and Backgrounds in Printed Acuity Tests for Infants and Young Children.
Acuity tests for infants and young children use preferential looking methods that require a perceptual match of brightness and color between grey background and target spatial average. As a first step in exploring this matching, this article measures photometric and colorimetric matches in these acuity tests. The luminance, uniformity, contrast, and color spectra of Teller Acuity Cards, Keeler Acuity Cards for Infants, and Lea Paddles under ambient, warm, and cold lighting, and of grey-emulating patterns on four digital displays, were measured. Five normal adults' acuities were tested at 10 m observationally. Luminance and spectral mismatches between target and background were found for the printed tests (Weber contrasts of 0.3% [Teller Acuity Cards], -1.7% [Keeler Acuity Cards for Infants], and -26% [Lea Paddles]). Lighting condition had little effect on contrast, and all printed tests and digital displays met established adult test luminance and uniformity standards. Digital display grey backgrounds had very similar luminance and color whether generated by a checkerboard, vertical grating, or horizontal grating. Improbably good psychophysical acuities (better than -0.300 logMAR: (logarithm of the minimum angle of resolution)) were recorded from adults using the printed tests at 10 m, but not using the digital test Peekaboo Vision. Perceptible contrast between target and background could lead to an incorrectly measured, excessively good acuity. It is not clear whether the luminance and spectral contrasts described here have clinically meaningful consequences for the target patient group, but they may be avoidable using digital tests. Current clinical infant acuity tests present photometric mismatches that may return inaccurate testing results.


Isolation and sequencing of a cDNA clone for a prolactin-inducible protein (PIP). Regulation of PIP gene expression in the human breast cancer cell line, T-47D.
We have initiated an analysis of the action of prolactin in a human breast cancer cell line (T-47D) by isolating and sequencing cDNA clones for a prolactin-inducible protein (PIP). PIP cDNA (approximately 600 base pairs) was isolated from a lambda gt11 expression library prepared from mRNA extracted from T-47D cells treated with prolactin and hydrocortisone. The identity of PIP cDNA was confirmed by hybrid-selected translation. PIP mRNA is a single mRNA species of approximately 900 bases which responds to lactogenic hormones (human prolactin and growth hormone) in a time- and dose-dependent manner. Treatment of T-47D cells with human growth hormone (hGH) increased PIP mRNA levels by 4.6-fold, while the combination of hGH and hydrocortisone or dihydrotestosterone had a more dramatic, 12.4-fold, effect. The latter steroid was the most potent. Dihydrotestosterone plus hGH increased transcription of the PIP gene by 3.7-fold. Dihydrotestosterone alone was as effective as dihydrotestosterone plus hGH in increasing the transcription rate, while hGH alone had no effect. Thus, lactogen may regulate PIP gene expression at a post-transcriptional step. PIP mRNA was expressed at low levels in other human breast cancer cell lines which were prolactin receptor-positive; MCF-7 and EFM-19 lines were exceptions. PIP cDNA had an open reading frame of 146 amino acids, sufficient to encode the precursor form of the secreted PIP protein (approximately 14.5 kDa). The similarity of the predicted amino acid sequence of PIP to the sequence of a protein encoded by a gene expressed in the mouse submaxillary gland prompted us to look for PIP in human saliva. Western blot analysis confirmed the presence of PIP in human saliva. Therefore, the PIP gene is useful in studying the molecular actions of the prolactin/growth hormone polypeptide hormone family and the interaction with androgen, in mammary and other potential target cells.


Body fat and its relationship with clustering of cardiovascular risk factors.
Body mass index (BMI) and waist circumference (WC) are the most commonly measured anthropometric parameters given their association with cardiovascular risk factors (RFs). The relationship between percentage body fat (%BF) and cardiovascular risk has not been extensively studied. This study evaluated %BF and its relationship with cardiometabolic RFs in healthy subjects and compared these findings with the relationship between BMI/ WC and cardiovascular RFs. This was a cross-sectional study of 99 males and 83 females (mean age 38 ± 10 years) evaluated in a preventive cardiology program. All subjects completed a survey about RFs and lifestyle habits. Anthropometric parameters, systolic blood pressure (SBP), diastolic blood pressure (DBP), fasting lipid profile, and blood glucose were collected. Body fat was determined using four skinfold measurements. Fat mass index (FMI) was also calculated. Percentage body fat was significantly and directly associated with total cholesterol (R(2)=0.11), triglycerides (R(2)=0.14), low-density lipoprotein cholesterol (R(2)=0.16), non-high-density lipoprotein cholesterol (R(2)=0.24), fasting blood glucose (R(2)=0.16), SBP (R(2)=0.22), and DBP (R(2)=0.13) (p<0.001 for all) and inversely related to high-density lipoprotein cholesterol (R(2)=0.32; p<0.001). When the models of %BF, FMI, WC, and BMI were compared, all of them were significantly related to the same cardiometabolic RFs and the clustering of them. Percentage body fat and FMI were significantly associated with biochemical variables and to the clustering of RFs. However, these associations were similar but not better than WC and BMI.


[Three-dimensional techniques in orthopedic ultrasonography].
Three-dimensional ultrasonography is not yet an established method in soft tissue and joint diagnostics as it is in obstetrics. Although a spatial view on the object is fascinating, problems occur in handling. No standard is defined for acquisition or visualisation. For the acquisition of the volume mainly 3D-arrays and image correlation techniques are used. For visualisation planar views of sections out of the volume, but also spatial views in a transparency or surface mode are used. Investigations on feasibility for soft tissue or joints show a higher accuracy, validity and reproducibility for 3D-sonography. Due to the low differences of the acoustic impedance visualisation is confusing in a transparency mode and needs a time consuming postprocessing in a surface mode. Future developments address to the field of faster acquisition ("real time 3D"), higher geometric precision and easier visualisation by automated segmentation. By the three-dimensionality of the data set applications in telematics, navigation and simulation ("sonographic training") are expected.


Heat induced aggregation of gold nanorods for rapid visual detection of lysozyme.
Gold nanorods have been nominated as propitious candidates for nanobiodiagnostic applications. Herein, a technique has been introduced for rapid visual detection of lysozyme, as its high level of excretion in biological fluids is a characteristic sign of leukemia and kidney disorders. Gold nanorods were biofunctionalized with lysozyme aptamer and characterized with UV-Visible and FTIR spectroscopy, zeta potential analyzer and transmission electron microscopy. Exposure of the nanoprobe to nano molar levels of lysozyme (20 nmol l(-1)) lead to dictated aggregation of the nanostructures at ambient temperature; which was significantly improved by heat induced morphological perturbations and rapid detection by the naked eye (down to pico molar level). Qualitative analysis of Acute myeloid leukemia, Acute lymphocytic leukemia and Lymphoma blood serums showed sensitivity and specificity of the fabricated aptasensor under both temperature conditions. This report encourages utilization of heat-induced aggregation of gold nanorods as a promising nanodiagnostic technique for the emerging nanotechnologies.


Peptidylarginine deiminase IV promotes the development of chemoresistance through inducing autophagy in hepatocellular carcinoma.
Peptidylarginine deiminase IV (PADI4) is widely distributed in several tissues and the expression is correlated with many pathological processes. Chemotherapy remains a major treatment alternatively to surgery for a large number of patients at the advanced stage of hepatocellular carcinoma (HCC). However, the role of PADI4 in the chemoresistance of HCC has not been identified. MTT and PI/Annexin V assay were employed to examine the proliferation and apoptosis of HCC cell lines. The expression of MDR1 is detected by Realtime PCR. GFP tagged LC3 expression vector and electron microscopy are utilized to demonstrate the occurrence of autophagy. We observed that the elevated PADI4 expression is associated with chemoresistance in HCC patients with TACE after surgery. In addition, we found that overexpression of PADI4 in HCC cell lines lead to the resistance to chemotherapeutic agents in vitro and in vivo. Interestingly, the HCC cells that overexpressed PADI4 were observed to undergo autophagy which was known as a protective mechanism for cells to resist the cell tosicity from chemotherapy. Autophagy inhibitor could effectively restore the sensitivity of HCC cells to chemotherapy in vitro and in vivo. These results indicate that PADI4 may induce chemoresistance in HCC cells by leading autophagy.


Isolated ventricular noncompaction is associated with coronary microcirculatory dysfunction.
We sought to analyze whether a microcirculatory dysfunction might be associated with isolated ventricular noncompaction (IVNC). In IVNC, which is a cardiomyopathy thus far "unclassified" by the World Health Organization, heart failure and sudden cardiac death are common findings, but the pathophysiologic mechanisms are unknown. In 12 patients with IVNC and 14 control subjects, quantitative evaluation of regional myocardial perfusion (myocardial blood flow [MBF]) and coronary flow reserve (CFR, hyperemic/baseline MBF) was performed using positron emission tomography and (13)N-ammonia. The left ventricular myocardium was divided into nine segments, and the two-dimensional echocardiogram in each patient with IVNC was compared with CFR in each segment. Noncompaction was defined as a two-layered structure with excessive trabeculation. The CFR in control subjects averaged 4.2+/-0.9, providing a cut-off value > or =2.5, but it was 2.1+/-0.8 in patients with IVNC. A perfusion scan defect was found in 14 of 24 segments with noncompaction, although no defect was found in 76 of 84 normal segments (overall agreement 83%, p < 0.0001 by the chi-square test). In 16 of 21 segments with noncompaction, a decreased CFR was found; but a decreased CFR was also found in 36 of 60 segments without noncompaction (p = NS). In 45 of the 57 segments with wall motion abnormalities, CFR was decreased, but it was preserved in 17 of the 24 segments with normal wall motion (agreement 77%, p < 0.0001). In patients with IVNC, a decreased CFR is not confined to noncompacted segments, but extends to most segments with wall motion abnormalities. Thus, coronary microcirculatory dysfunction is associated with IVNC.


Decreased blood pressure with a corresponding decrease in adhesive molecules in diabetic rats caused by vitamin E administration.
Hypertension is one of the important clinical problems of diabetic cardiovascular disease. The aim of this study was to determine the effect of vitamin E on blood pressure parameters and adhesive molecule amounts in diabetic rats. Twenty-four male Wistar rats were divided into three groups (each of n = 8): the controls (C), non-treated diabetic (NTD), and vitamin E treated diabetic (VETD) groups. A single intraperitoneal injection of buffered streptozotocin (60 mg/kg) in cold sodium citrate (pH 4.5) was used to induce diabetes. The VETD group received 300 mg of vitamin E daily intragastrically for 6 weeks. Systolic and diastolic blood pressure, mean arterial pressure, as well as the dicrotic pressure, crest time, systolic and diastolic periods, and plasma levels of intercellular adhesion molecule-1 and E-selectin were measured after 6 weeks. The results revealed that there was a significant increase in systolic and diastolic blood pressures, mean arterial pressure, crest time, systolic duration, and the amount of sICAM-1 and E-selectin in diabetic rats. There was no significant difference in the heart rate or cardiac cyclic duration among the different groups. Significant improvement of blood pressure parameters as well as attenuation of the elevated ICAM-1 and E-selectin amounts was found in the vitamin E treated group. These findings indicate that vitamin E significantly improved blood pressure elevation in diabetic rats and that these effects could be associated with reducing adhesive molecule and antioxidant properties of vitamin E.


Combination effects of chronic cadmium exposure and gamma-irradiation on the genotoxicity and cytotoxicity of peripheral blood lymphocytes and bone marrow cells in rats.
This work investigated the effects of chronic cadmium (Cd) exposure combined with γ-ray irradiation on the cytotoxicity and genotoxicity of peripheral blood cells and bone marrow cells in rats. Results showed that when the rats were exposed to low dose (LD) Cd of 0.1mg CdCl₂/(kgd) for 8 and 12 weeks, the Cd concentration in blood reached to 135-140 μg/L and no toxic effects on peripheral blood lymphocytes, white blood cells (WBC) and granulocyte-monocyte (GM) progenitor cells were observed except polychromatic erythrocytes (PCE) of bone marrow. Moreover, this chronic LD Cd exposure significantly decreased irradiation-induced micronucleus (MN) formation and hypoxanthine-guanine phosphoribosyl transferase (hprt) mutation in lymphocytes and PCE, while the combination of LD Cd exposure and irradiation induced the additive metallothionein (MT) protein expression in bone marrow cells. When the rats were exposed to a high dose (HD) Cd of 0.5mg CdCl/₂(kgd) for 8 and 12 weeks, the blood Cd level approached to 458-613 μg/L and an inflammatory response was induced, meanwhile, MN formation and hprt mutation were markedly increased, and the ratio of PCE/NCE (normochromatic erythrocyte) was significantly decreased. Furthermore, when the rats were exposed to HD Cd plus 2 Gy irradiation, additive toxic effects on MN formation, hprt mutation, PCE damage and GM progenitor cell proliferation were observed, while this combination treatment resulted in an obvious reduction of MT protein compared to HD Cd group. In conclusion, chronic exposure to LD Cd induced the adaptive response to irradiation in the genotoxicity of peripheral blood lymphocytes and PCE of bone marrow by the up-regulation of Cd-induced MT protein, but the combination of HD Cd exposure and irradiation generated the additive effects on the cytotoxicity and genotoxicity in peripheral blood lymphocytes and bone marrow cells.


Congenital diaphragmatic hernia in Smith-Magenis syndrome: a possible locus at chromosome 17p11.2.
We report on a 7-month-old girl with Smith-Magenis syndrome (SMS) due to a 4.76-Mb deletion of 17p12-17p11.2 detected by array comparative genomic hybridization. She was also affected with a left-sided congenital diaphragmatic hernia (CDH) and cardiac anomalies including an atypical atrioventricular canal defect and a cleft mitral valve. To our knowledge, this is the first reported case of a patient with both SMS and CDH. There are numerous chromosomal regions in which duplications, deletions, inversions, or translocations have been associated with CDH, but none have previously been reported at or close to 17p11.2. We discuss candidate genes for the diaphragmatic defect in this patient. Our case demonstrates that it is important to consider the possibility of SMS in non-isolated cases of diaphragmatic hernia.


Mechanical thrombectomy of hemodialysis fistulae and grafts.
In this article, the authors present approaches they use in performing dialysis access intervention-in particular clotted access. It is not meant to be a comprehensive review of dialysis access management. At our institution, mechanical thrombectomy is the primary mode of treatment for clotted hemodialysis access. We will present physical examination findings in clotted dialysis access and contraindications for mechanical thrombectomy in dialysis access. We will also discuss the devices for mechanical thrombectomy and the techniques we use. Finally, we will discuss the difficulties encountered in these procedures and their solutions.


Radiological signs of a left atrial aneurysm.
We report a case of an acquired mid-sized extrapericardial aneurysm in the area of the left atrial appendage and demonstrate the diagnostic impact of several imaging methods. Chest radiography gave a hint to the diagnosis with a bulky mass of soft tissue density appearing adjacent to the left atrial appendage. Transthoracic as well as transesophageal echocardiography missed the origin of the aneurysm. Magnetic resonance imaging helped to establish the diagnosis with use of a coronal imaging plane. No contrast agent was necessary. Thus, the combination of chest X-ray and MRI provided excellent support for patient management.


Duplicate casts in fabrication of temporary removable partial dentures.
The use of blocked-out duplicate casts in the fabrication of temporary removable partial dentures has been described. Temporary removable partial dentures produced in this controlled manner fit properly and aid in minimizing intraoral damage. Accuracy of fit is assured because all relief is accomplished by block out on the surveyor during the laboratory phase rather than by random grinding of the prosthesis at insertion. In addition, the original cast is salvaged and can be used as a meaningful reference during the finishing process in the laboratory (Fig. 5).


A simple method for ranking the affinities of monoclonal antibodies.
Measurement of the binding of constant trace amounts of labelled antigen by increasing dilutions of culture supernatant allows the ranking of monoclonal antibodies in the order of their affinity for the antigen. The theoretical basis for this method is discussed and it is illustrated with data from a set of anti-alphafoetoprotein monoclonal antibodies. Hybridomas secreting antibodies of desired affinity for immunoassay, histochemistry or antigen purification can thus be selected at an early stage after fusion.


Clinical indications of the use of antineutrophil cytoplasmic antibodies and anti-Saccharomyces cerevisiae antibodies in the evaluation of inflammatory bowel disease at an Academic Medical Center.
Serological markers for inflammatory bowel disease (IBD), including anti-neutrophil cytoplasmic antibody (ANCA) and anti-Saccharomyces cerevisiae antibody (ASCA), have a high specificity and positive predictive value in diagnosing IBD. However, neither indication nor use in clinical practice has been clearly established. This study aimed to determine the clinical impact of ANCA/ASCA testing by evaluating how these tests were used in an academic referral center. Retrospective chart review to classify the indications for testing and effect on diagnosis or management. Seventy-six patients had serological tests for IBD. Indications included differentiating ulcerative colitis (UC) from Crohn's disease (CD) in established patients with IBD (13%); establishing a diagnosis in patients with atypical signs of inflammation as detected by endoscopy, histology, or radiology (50%); evaluation of chronic diarrhea (22%); evaluation of a family history of IBD (4%); and differentiating pouchitis from CD (4%). Review of the subsequent course indicated that serologic testing had an important role in diagnosis in 28% of patients, a supportive role in 26%, and was not helpful in 46%. Serologic testing clarified the clinical presentation in 61% of those presenting with atypical inflammatory changes. It proved valuable in establishing a diagnosis of UC or CD in a subset of middle-aged patients with inflammatory changes in the sigmoid colon. For patients with chronic diarrhea, the yield was lower: 36% had a significant effect on diagnosis, but test results changed immediate treatment in only 1 (6%). In patients being considered for operative management (n = 8), serologic testing was valuable in clarifying the diagnosis in 75% of patients and had an impact on the operative plan in 62% of patients. Serological testing for ANCA/ASCA may have a significant role in the diagnosis and treatment in individuals presenting with sigmoid inflammation or atypical inflammation but was less useful in those with chronic diarrhea.


Intermingling of gut microbiota with brain: Exploring the role of probiotics in battle against depressive disorders.
Depression is a debilitating psychiatric ailment which exerts disastrous effects on one's mental and physical health. Depression is accountable for augmentation of various life-threatening maladies such as neurodegenerative anomalies, cardiovascular diseases and diabetes. Depressive episodes are recurrent, pose a negative impact on life quality, decline life expectancy and enhance suicidal tendencies. Anti-depression chemotherapy displays marked adverse effects and frequent relapses. Thus, newer therapeutic interventions to prevent or combat depression are desperately required. Discovery of gut microbes as our mutualistic partner was made a long time ago and it is surprising that their functions still continue to expand and as of yet many are still to be uncovered. Experimental studies have revealed astonishing role of gut commensals in gut-brain signaling, immune homeostasis and hormonal regulation. Now, it is a well-established fact that gut microbes can alleviate stress or depression associated symptoms by modulating brain functions. Here in, we provide an overview of physiological alleyways involved in cross-talk between gut and brain, part played by probiotics in regulation of these pathways and use of probiotic bacteria as psychobiotics in various mental or depressive disorders.


Ultrasensitive detection of scrapie prion protein derived from ARQ and AHQ homozygote sheep by interspecies in vitro amplification.
Prions, infectious agents causing TSEs, are composed primarily of the pathogenic form (PrP(Sc)) of the PrP(C). The susceptibility of sheep to scrapie is determined by polymorphisms in the coding region of the PRNP, mainly at codons 136, 154, and 171. The efficiency of in vitro amplification of sheep PrP(Sc) seems to be linked also to the PrP genotype. PrP(Sc) derived from sheep with V(136)R(154)Q(171)-associated genotypes can be amplified efficiently by PMCA in the presence of additional polyanion such as poly A, but there are no reports that cite ultrasensitive detection of PrP(Sc) derived from sheep of other PrP genotypes. We report here that sheep PrP(Sc) derived from ARQ and AHQ homozygotes was amplified efficiently by serial PMCA using mouse brain homogenate as PrP(C) substrate. ARQ/ARQ PrP(Sc) was detected in infected brain homogenates diluted up to 10(-10) after five rounds of amplification, and AHQ/AHQ PrP(Sc) was detected in samples diluted up to 10(-8) after four rounds of amplification. On the other hand, amplification of PrP(Sc) from VRQ/ARQ sheep seemed to be less efficient under the experimental conditions used. The interspecies PMCA developed in this study may be useful in the detailed analysis of PrP(Sc) distribution in classical scrapie-infected ARQ and AHQ homozygote sheep.


Structural basis for exon recognition by a group II intron.
Free group II introns are infectious retroelements that can bind and insert themselves into RNA and DNA molecules via reverse splicing. Here we report the 3.4-A crystal structure of a complex between an oligonucleotide target substrate and a group IIC intron, as well as the refined free intron structure. The structure of the complex reveals the conformation of motifs involved in exon recognition by group II introns.


Polymeric alkenoxy amino acid surfactants: III. Chiral separations of binaphthyl derivatives.
Micellar electrokinetic chromatography (MEKC) was investigated for the enantiomeric separations of three binaphthyl derivatives ((+/-)-1,1'-bi-(2-naphthol) (BOH), (+/-)-1,1'-binaphthyl-2,2'-diyl hydrogenphosphate (BNP), and (+/-)-1,1'-binaphthyl-2,2'-diamine (BNA)) using two recently synthesized chiral polymeric surfactants (polysodium N-undecenoxy carbonyl-L-leucinate (poly-L-SUCL) and polysodium N-undecenoxy carbonyl-L-isoleucinate (poly-L-SUCIL)) in our laboratory. Enantiomeric separation (resolution and selectivity) of the binaphthyl derivatives was influenced by polymerization concentration of the monomeric surfactant, pH, type and concentration of the background electrolyte (BGE) as well as concentration of the polymeric surfactant. Two BGEs (dibasic phosphate and Tris-borate) were compared for this study. The use of dibasic phosphate as BGE in poly-L-SUCL provides baseline resolution of (+/-) BOH and (+/-) BNP, however, no resolution and selectivity at all was observed for (+/-) BNA. A similar approach was adopted with Tris-borate-poly-L-SUCL system at fixed pH 10.1, which resulted in baseline resolution of all three binaphthyl derivatives. Although R(s) of binaphthyl derivatives was always higher and electroosmotic flow (EOF) was always lower using Tris-borate than with dibasic phosphate, the selectivity values for the two buffer systems did not differ significantly. In addition, it was found that poly-L-SUCL provided better enantiomeric resolution and selectivity for (+/-) BOH and (+/-) BNA, while poly-L-SUCIL provided enhanced enantiomeric resolution but similar enantioselectivity for (+/-) BNP. This indicates that the depth of analyte penetration into the palisade layer and the micellar core are responsible for chiral recognition of hydrophobic analyte (e.g., (+/-) BOH, and (+/-) BNA) whereas for moderately hydrophobic analyte (e.g., (+/-) BNP) interaction with the polar head group seems to dictate chiral recognition. Simultaneous enantioresolution of all three binaphthyl derivatives was possible in a single electrophoretic run using either poly-L-SUCL or poly-L-SUCIL. Further comparison of the two polymeric surfactants showed that poly-L-SUCL provided slightly longer analysis time than poly-L-SUCIL but the use of the former polymeric surfactant should be preferred due to its ability to provide complete baseline resolution and higher selectivity of all the three atropisomers with a wider chiral window.


Protein oxidation in aging and the removal of oxidized proteins.
Reactive oxygen species (ROS) are generated constantly within cells at low concentrations even under physiological conditions. During aging the levels of ROS can increase due to a limited capacity of antioxidant systems and repair mechanisms. Proteins are among the main targets for oxidants due to their high rate constants for several reactions with ROS and their abundance in biological systems. Protein damage has an important influence on cellular viability since most protein damage is non-repairable, and has deleterious consequences on protein structure and function. In addition, damaged and modified proteins can form cross-links and provide a basis for many senescence-associated alterations and may contribute to a range of human pathologies. Two proteolytic systems are responsible to ensure the maintenance of cellular functions: the proteasomal (UPS) and the lysosomal system. Those degrading systems provide a last line of antioxidative protection, removing irreversible damaged proteins and recycling amino acids for the continuous protein synthesis. But during aging, both systems are affected and their proteolytic activity declines significantly. Here we highlight the recent advantages in the understanding of protein oxidation and the fate of these damaged proteins during aging. This article is part of a Special Issue entitled: Posttranslational Protein modifications in biology and Medicine.


Novel Surgical Strategy for Treating Osteoporotic Vertebral Fractures with Cord Compression.
Treatment for osteoporotic vertebral fracture (OVF) with cord compression is challenging and it usually requires surgical interventions to decompress nerves and restore spinal sequences. To describe a novel surgical strategy for treating OVFs with cord compression. This is a single-center retrospective analysis. The inclusion criteria were Frankel grade C-E, single level T10 -L2 . Between January 2008 and December 2016, a total of 56 OVF patients (47 females and nine males, with an average age of 72 years (66-88 years), comprising of eight grade C, 23 grade D, and 25 grade E patients) were enrolled. The treatment algorithm included preoperative evaluation by MRI, extension CT, and radiography to classify the OVFs as type 1.1 (reducible, stable; n = 13), type1.2 (reducible, unstable; n = 16), type 2 (irreducible; n = 19) or type 2M (modifier; n = 8). Vertebroplasty (VP)/kyphoplasty (KP) was applied in type 1.1. VP/KP with posterior fixation and posterolateral fusion was applied in type 1.2. And additional laminectomy/osteotomy was used in type 2, except in a modifier group (2M) where same procedure as applied for type 1.2 was used. VAS, ODI, Cobb angle, Frankel functional grade, and complications were recorded. Thirteen cases were classified as type 1.1, 16 cases as type 1.2, 19 cases as type 2, and eight cases as type 2M. The follow-up period was 38.9 months (range, 24-108 months). All patients were followed-up in at least 24 months, in which time four patients died, two patients were lost at the last follow-up, and 50 patients completed the full study. The total VAS and ODI improved from 8 (7, 9) and 75.5% (67.2%, 80.0%) preoperatively to 2 (1, 3) and 31% (24.0%, 37.0%) on conclusion, respectively (P < 0.01). The local kyphotic angle was corrected from 22.3° (17.1°, 33. 8°) preoperatively to 10.4° (6.4°, 15.3°) on conclusion (P < 0.01). Twenty-three patients had achieved neurological recovery on conclusion (42E, 8D, P < 0.01). Asymptotic cement leakage was observed in 17/56 cases (30.4%), 6/56 in the affected vertebra (10.7%), and 24/330 in the screw trajectory (7.3%). At 2 years postoperatively, 11 new VFs had occurred in nine patients (16.1%), including VFs in nine adjacent segments that all occurred within 1 year after surgery. No cement migration or implant failure was noted. The novel surgical strategy for treating OVFs with cord compression consists of the most tailored and least invasive treatment for each patient. The positive mid- and long-term clinical and radiological outcomes observed could represent a step forward in devising the proposed algorithm.


Nanocrystalline spherical hydroxyapatite granules for bone repair: in vitro evaluation with osteoblast-like cells and osteoclasts.
Conventionally sintered hydroxyapatite-based materials for bone repair show poor resorbability due to the loss of nanocrystallinity. The present study describes a method to establish nanocrystalline hydroxyapatite granules. The material was prepared by ionotropic gelation of an alginate sol containing hydroxyapatite (HA) powder. Subsequent thermal elimination of alginate at 650 °C yielded non-sintered, but unexpectedly stable hydroxyapatite granules. By adding stearic acid as an organic filler to the alginate/HA suspension, the granules exhibited macropores after thermal treatment. A third type of material was achieved by additional coating of the granules with silica particles. Microstructure and specific surface area of the different materials were characterized in comparison to the already established granular calcium phosphate material Cerasorb M(®). Cytocompatibility and potential for bone regeneration of the materials was evaluated by in vitro examinations with osteosarcoma cells and osteoclasts. Osteoblast-like SaOS-2 cells proliferated on all examined materials and showed the typical increase of alkaline phosphatase (ALP) activity during cultivation. Expression of bone-related genes coding for ALP, osteonectin, osteopontin, osteocalcin and bone sialoprotein II on the materials was proven by RT-PCR. Human monocytes were seeded onto the different granules and osteoclastogenesis was examined by activity measurement of tartrate-specific acid phosphatase (TRAP). Gene expression analysis after 23 days of cultivation revealed an increased expression of osteoclast-related genes TRAP, vitronectin receptor and cathepsin K, which was on the same level for all examined materials. These results indicate, that the nanocrystalline granular materials are of clinical interest, especially for bone regeneration.


Experiences of inpatient eating disorder admissions: A systematic review and meta-synthesis.
There has been a significant increase in the number of inpatient admissions for individuals with eating disorders and, with the most critical cases requiring inpatient treatment, it is essential that we continue to improve associated outcomes. The aim of the study was to synthesise the available qualitative literature on the experiences of inpatient admissions for eating disorders to understand individuals' experiences and identify areas that may require further research and/or service development. Searches were performed on the following online databases: PsycINFO, PsycArticles, PsycTherapy MEDLINE, Embase, CINAHL, ASSIA, Scopus and Proquest Open Access Theses. Only papers with qualitative data regarding individuals' experiences of inpatient eating disorder treatment were considered. The CASP qualitative checklist was used to assess studies and relevant data items were extracted. Thematic synthesis was used to integrate the findings in the identified studies. GRADE-CERQual was used to rate the confidence in the findings. Twenty-eight studies were identified which the CASP assessment considered to be adequate. The synthesis produced 5 main themes; 'Care and control', 'Inpatient bubble', 'Being supported and understood', 'Challenges of living with others' eating disorders' and finally 'Relationship to eating disorder'. The GRADE CERQual framework rated findings with high or moderate confidence. Findings reaffirmed the importance of patient-centred care and the significant impact of being separated from normal life with others also experiencing an eating disorder.


Fluorous-assisted metal chelate affinity extraction technique for analysis of protein kinase activity.
We have developed a fluorous affinity-based extraction method for measurement of protein kinase activity. In this method, a fluorescent peptide substrate was phosphorylated by a protein kinase, and the obtained phosphopeptide was selectively captured with Fe(III)-immobilized perfluoroalkyliminodiacetic acid reagent via a metal chelate affinity technique. Next, the captured phosphopeptide was selectively extracted into a fluorous solvent mixture, tetradecafluorohexane and 1H,1H,2H,2H-tridecafluoro-1-n-octanol (3:1, v/v), using the specificity of fluorous affinity (fluorophilicity). In contrast, the remained substrate peptide in the aqueous (non-fluorous) phase was easily measured fluorimetrically. Finally, the enzyme activity could be assayed by measuring the decrease in fluorescence. The feasibility of this method was demonstrated by applying the method for measurement of the activity of cAMP-dependent protein kinase (PKA) using its substrate peptide (kemptide) pre-labeled with carboxytetramethylrhodamine (TAMRA).


Prevention of recurrent urinary tract infections in premenopausal women by post-coital administration of cinoxacin.
A total of 21 sexually active premenopausal women, prone to recurrent urinary tract infections but who otherwise were healthy, underwent post-coital prophylaxis consisting of bladder voiding and a single 250 mg. tablet of cinoxacin. While 94 urinary tract infections occurred during a mean of 7.5 months before treatment, only 8 occurred during a mean of 12.5 months after prophylaxis. This difference was statistically highly significant. A mean of 106 cinoxacin tablets per patient were administered during post-coital prophylaxis. Cinoxacin represents an additional valuable and effective antibacterial in post-coital prophylaxis of recurrent urinary tract infection in otherwise healthy premenopausal women, although it is slightly less effective than cotrimoxazole or nitrofurantoin. Effective post-coital prophylaxis requires the use of much smaller quantities of antibacterial agents than the daily use of a single tablet and in women with a high incidence of recurrent urinary tract infections it is superior to intermittent self-administered antibacterial therapy.


NLRP12: The Monarch of the Gut.
Innate pattern recognition receptors have been implicated in the obesity-associated imbalance of gut microbiota. In this issue of Cell Host & Microbe, Truax et al. (2018) report that NLRP12 prevents high-fat-diet-induced obesity by maintaining beneficial short-chain fatty acid-producing microbiota.


Oocyst counts in crossbred ewes under tree-crop plantation in the forest zone of Ghana.
In this paper, the Eimeria oocyst output of two groups, pregnant ewes (group 1) and non-pregnant controls (group 2), which were followed from September 1993 to August 1994, is described. In both groups of animals the level of oocyst output was high during the minor rainy season. However, during the periparturient period the pregnant ewes showed the higher oocyst output. The oocyst output in both group fell to similar levels after weaning of the lambs in March 1994. The species of Eimeria identified in order of dominance were Eimeria parva, E. pallida, E. faurei, E. ahsata, E. ovina, E. intricata, E. granulosa and E. ninakohlyakimovae. There were no differences in the species composition of oocysts in both groups of animals.


The effect of endoscopic ultrasound on the precise selection of endoscopic treatment for submucosal tumors in the upper gastrointestinal tract.
To summarize and discuss the guiding role of endoscopic ultrasound (EUS) in selecting endoscopic treatments for submucosal tumors (SMTs) in the upper gastrointestinal tract. A retrospective investigation was conducted on 156 SMT patients who received endoscopic resection guided by EUS in the endoscopy center of the Second Affiliated Hospital of Guangzhou University of Chinese Medicine from May 2019 to September 2021. Next, the size, pathological type, and distribution of lesions were analyzed; the correlation of the tumor origin with distribution of lesions and selection of treatments was explored; and the consistency of preoperative EUS diagnosis and postoperative pathological diagnosis was summarized and analyzed. The tumor diameters of the included SMT patients ranged from 0.3 to 4 cm, with a mean diameter of 0.95 cm; the lesions were mostly located in the esophagus, gastric fundus or fundic cardia and gastric body. As for the pathological types, liomyoma was the most common tumor in the esophagus, liomyoma and mesenchymoma were mainly located in the fundic cardia and gastric body, and heterotopic pancreas was mostly discovered in the gastric sinus. Among 38 esophageal SMT patients, some with lesions originating from muscularis mucosa and submucosa under EUS mainly underwent endoscopic submucosal dissection (ESD) and endoscope band ligation (EBL); while others with lesions originated from muscularis propria mainly received submucosal tunneling endoscopic resection (STER). Of 115 gastric SMT patients under EUS, some with lesion origins from the muscularis mucosa and submucosa mainly underwent endoscopic submucosal excavation (ESE), while others from muscularis propria mainly underwent ESE, ESD, and endoscopic full-thickness resection (EFTR). Besides, 3 duodenal SMT patients with lesion origins from submucosa and muscularis propria under EUS were given ESD and ESE, respectively. Additionally, 121 cases showed a consistency between the EUS diagnosis and the postoperative pathological nature, and the consistency rate was 84.6%. Clarifying the origin layer, size, growth pattern, and pathological nature of the lesion through preoperative EUS can guide the precise selection of endoscopic treatments, thereby ensuring a safe, effective, and complete surgical outcomes and reducing complications.


Synthesis of C17-OH-north unit of ritterazine G via "Red-Ox" modifications of hecogenin acetate.
The C17-OH-north unit of ritterazine G was prepared in 13 steps from hecogenin acetate. This synthesis features a highly efficient and stereoselective introduction of the C17-OH via E-ring cleavage/F-ring formation, D-ring oxidation, and F-ring cleavage/E-ring formation.


A BODIPY-based fluorescent probe for ratiometric detection of gold ions: utilization of Z-enynol as the reactive unit.
Using an irreversible intramolecular cyclisation pathway triggered by gold ions, a boron-dipyrromethene (BODIPY) based fluorescent probe integrated with a reactive Z-enynol motif responds selectively to gold ions. With the addition of gold(iii), the probe displays ratiometric fluorescence behaviour clearly observable to the naked eye under both visible and UV light.


Determination and pharmacokinetics of acyclovir after ingestion of suspension form.
The study describes, simple, precise, sensitive and accurate HPLC assay with spectrofluorimetric detection for the determination of acyclovir in human plasma. The method was linear over a range 25 1200 ng ml(-1). The average yield in this method exceeded 80%. Limits of quantitation and detection were 25 and 10 ng ml(-1), respectively. On the basis of reported method, a single-dose of pharmacokinetics on 24 men, in two doses (200 and 400 mg) of acyclovir suspension has been investigated. Pharmacokinetic parameters obtained from both doses of the drug were compared. The linearity of acyclovir pharmacokinetics in the investigated dose ranges has been confirmed.


Efficacy of Transcutaneous Electrical Nerve Stimulation for Postoperative Pain, Pulmonary Function, and Opioid Consumption Following Cardiothoracic Procedures: A Systematic Review.
Transcutaneous electrical nerve stimulation (TENS) is a minimally invasive method for treating pain. In the most recent review published in 2012, TENS was associated with increased pain relief following cardiothoracic surgery when compared to standard multimodal analgesia. The purpose of this systematic review and meta-analysis is to determine if adding TENS to current pain management practices decreases pain and analgesic use and improves pulmonary function for postcardiothoracic surgery patients. CINAHL, MEDLINE, Cochrane Database of Systematic Reviews, PubMed, and ClinicalTrials.gov were searched using specific keywords. Covidence was used to screen, select studies, and extract data by two independent reviewers. The Cochrane Risk of Bias tool assessed risk of bias. Visual analog scale (VAS) and pulmonary function data were exported for meta-analysis using a random effects model. The search yielded 38 articles. Eight randomized controlled trials met inclusion criteria for the literature review. Five studies were included in the meta-analysis of pain at 24, 48, and 72 hours postoperatively. Data were analyzed using the standard mean difference (SMD). TENS had a significant impact on VAS at rest (-0.76 SMD [95% confidence interval, CI = -1.06 to -0.49], p < 0.00001) and with coughing (-1.11 SMD [95% CI = -1.64 to -0.56], p < 0.0001). FEV1 improved after 72 hours (1.00 SMD [95% CI = 0.66-1.35], p < 0.00001), as did forced vital capacity (1.16 SMD [95% CI = 0.23-2.10], p = 0.01). The addition of TENS therapy to multimodal analgesia significantly decreases pain following cardiothoracic surgery, increases the recovery of pulmonary function, and decreases the use of analgesics.


Plasma lipoprotein changes in suckling and weanling rabbits fed semipurified diets.
The distribution and composition of the plasma lipoproteins were studied in suckling rabbits and in rabbits either weaned to or fed as young adults cholesterol-free, semipurified diets containing protein of animal (casein) or plant (soy protein isolate) origin. The raised cholesterol levels of the suckling period resulted in an increase of very low density and intermediate density lipoproteins in a making similar to that seen in adult rabbits fed a high fat diet supplemented with cholesterol. Young rabbits fed the cholesterol-free, semipurified, diet containing casein also became hypercholesterolemic but, in this case, the increased cholesterol was found primarily in lipoproteins of intermediate density. Cholesterol levels remained low in those rabbits fed the soy protein isolate diet, and the lipoprotein distribution was somewhat similar to that of chow-fed animals. It appears that the raised cholesterol levels during the suckling period result in different lipoprotein patterns to those produced in young adult animals by a cholesterol-free, semipurified diet.


Identification of a potent analogue of Nazumamide A through iteration of combinatorial tetrapeptide libraries.
Five sets of N-acylated tetrapeptide libraries and sublibraries related to Nazumamide A have been prepared using 25 natural and unnatural amino acids. They were evaluated in antithrombin assay, in order to quantify inhibition at each step of the tetrapeptide sublibrary iteration. The studies led to the identification of 2,5-dihydroxybenzoyl-lysyl-isoleucyl-phenylalanyl-arginine as a novel inhibitor of thrombin and was found to be at least 25 times more potent than the natural tetrapeptide 2,5-dihydroxybenzoyl-arginyl-prolyl-isoleucyl-alpha-aminobutyric acid (NAZA).


Intermittent occlusion of the abdominal aorta: a conceptual approach in treating below-diaphragm tumors (a preliminary report).
A modified form of tumor strangulation as a possible treatment modality is considered. Intermittent intra-aortic occlusion by means of specially designed catheters and equipment is described. As a preliminary feasibility study, occlusion in non-tumor-bearing animals has begun, with projection for future human cases anticipated. Theoretical concepts of substances from the occlusion process resulting in cure and specific immunity in experimental animals are evaluated, and the belief in the development of modified antigens and antibodies as a consequence of restriction of a growing tumor's proliferating blood supply is discussed.


Severity of disability related to road traffic crashes in the Spanish adult population.
The severity of disability related to road traffic crashes has been little studied, despite the significant health and socio-economic impacts that determine victims' quality of life. To estimate the consequences of road traffic crashes on the severity of disability, in terms of individuals' capacity to execute activities and perform tasks in their current environment, using aids. Cross-sectional study conducted on community-dwelling participants in the "2008 National Survey of Disability", with data on 91,846 households having 20,425 disabled persons, 443 of whom had disability due to road traffic crashes. We measured severity using two indicators, i.e., the Capacity (CSI) and Performance (PSI) Severity Indices. The highest proportion of disability was mild (CSI=70.5%; PSI=80.8%), while 7.6% (CSI) and 4.9% (PSI) was severe/complete respectively. The moderate/severe disability rate was 0.6 per thousand on the CSI, decreasing to 0.4 per thousand on the PSI. No differences were observed by age or sex. Moderate/severely disabled persons had a fourfold higher probability of being retired or unfit for work. Mental and nervous system impairments were more closely related to moderate/severe/complete problems of capacity and performance (p<0.001), disability for carrying out general tasks and demands, and interpersonal interactions and relationships (p<0.001). Being permanently bedridden (p<0.001), receiving aids (p<0.001), family support (p<0.05) and moving home (p<0.05) increased with an increase in the level of severity. Road traffic crashes mainly cause mild disability. Moderate/severe disability is associated with lower work capacity, greater functional dependence, and increased need of aids, moving home and family support.


Fibronectin alters the phenotypic properties of cultured chick embryo chondroblasts.
The state of chick embryo chondroblasts in culture was found to be sensitive to both fibronectin and another substance(s) (activity A) which could be extracted from chick embryo fibroblasts with 1 M urea or from conditioned medium. In the presence of either of these activities at concentrations of 25-150 micrograms/ml, chondroblasts, which normally grow as mixed cultures of floating and adherent cells, all immediately became attached to the tissue culture dish and spread. After several days, the morphology of these typically epithelioid cells became fibroblastic. This did not involve a selection process, since the effect was reversible. The synthetic program of these cells was also dramatically modified: the cultures no longer synthesized the chondroblast-unique type IV sulfated proteoglycan and began synthesizing alpha 2 collagen chains typical of fibroblastic or early limb bud cells. Fibronectin was resolved from activity A by gelatin affinity chromatography or gel filtration. Both activities were trypsin-sensitive. The two activities differed, however, on the basis of how the protein fractions in which they were found migrated in SDS-polyacrylamide gels, their specific activities and their effects on cell morphology and cell growth.


Effect of hydrophobicity of acyl groups on the activity and stability of acylated thermolysin.
1. Normal carboxylic acids of different hydrophobicities and similar chain lengths were prepared and used for the modification of amino groups of thermolysin (EC 126.96.36.199). They were 4,7,10,13-tetraoxatetradecanoic acid, 4,7,10-trioxatetradecanoic acid, 4,7-dioxatetradecanoic acid and 4-oxatetradecanoic acid. 2. The modified enzymes were isolated by gel filtration. They had 6--7 acyl groups per molecule. Acylation of amino groups with 4-oxatetradecanoic acid and tetradecanoic acid made the enzyme insoluble. 3. The most hydrophilic enzyme derivative had similar enzyme activity and higher heat resistance than the native enzyme. The most hydrophobic derivative showed lower Km (50%) and V (40%) values for proteinase activity and lower heat resistance than the former derivative. The trioxa-derivative had intermediate characteristics. The results are discussed with respect to effects on stability and activity of the enzyme.


Irradiation of murine lip mucosa in combination with 5-fluorouracil, administered by single dose injection or continuous infusion.
The possible interaction of 5-fluorouracil (5-FU) and irradiation on mouse lip mucosa was studied, with special interest for the influence of the route of administration of the drug, either by single intraperitoneal (i.p.) injection or by 7 days continuous subcutaneous infusion. Apart from the possible modification by the drug of radiation-induced damage to the lip mucosa assessment was made of the systemic toxicity. No modification of the radiosensitivity of mouse lip mucosa nor of its repair capacity was observed with the use of 5-FU given either by i.p. injection or by continuous infusion. However, the combination of 5-FU and irradiation resulted in an increased systemic toxicity which was only seen when 5-FU was administered by an i.p. injection in close timing with irradiation. It is suggested that this increased systemic toxicity is caused by the combined effects of irradiation and 5-FU on the alimentary tract, although we could not demonstrate the mechanism of it.


Guidelines on Hair Restoration for East Asian Patients.
In East Asian countries, hair transplantation is a quite common procedure for treating pattern hair loss, cosmetically correcting the hairline, and correcting eyebrow and pubic hair defects. Although there are general guidelines concerning hair transplantation, certain factors need to be addressed to make the guidelines more specific and suitable to East Asian requirements. To provide guidelines for hairline design, donor harvesting, graft preparation and placement, and medical treatment that are appropriate for hair transplantation in East Asian patients. Recommendations are based on the experience of the authors, surgeons who perform hair transplantation, and a comprehensive review of the available literature on hair transplantation in East Asians. Data on hair thickness and graft density, hairline design, and graft creation and placement techniques have been collaboratively evaluated and used to establish overall guidelines. The use of the proposed guidelines by surgeons will hopefully enhance outcomes and bring greater consistency to hair transplantation procedures for East Asian patients.


Cerebral blood flow in patients with Fabry disease as measured by Doppler sonography is not different from that in healthy individuals and is unaffected by treatment.
The purpose of this study was to investigate cervical and cerebral blood flow characteristics in patients with Fabry disease at baseline and under enzyme replacement therapy. In this case-control study we prospectively studied 68 patients with Fabry disease with extracranial and transcranial Doppler sonography. We compared extracranial and transcranial cervical and cerebral blood flow properties in all patients with Fabry disease and in subgroups of those with or without enzyme replacement therapy, male and female, and with normal or impaired renal function. Eight male patients were investigated at baseline and 1 year after initiation of enzyme replacement therapy. We show that cervical and cerebral blood flow parameters in patients with Fabry disease are not different from normal values regardless of sex, renal function, or enzyme replacement therapy. Cervical and cerebral blood flow measured with extracranial and transcranial Doppler sonography is not altered in patients with Fabry disease. Enzyme replacement therapy does not change blood flow characteristics.


Quantification of methotrexate in plasma samples using highly fluorescent nanoparticles.
A novel amine-functionalized silica carbon polymer dots (APTES-CPDs)-based sensor was developed for methotrexate (MTX) detection. APTES-CPDs core-shell structure was synthesized via a one-step hydrothermal process and were employed as an optical sensor for MTX determination due to quenching effect of MTX on the fluorescence intensity of CPDs. Under optimum conditions, the developed sensor detects MTX in plasma samples in the dual linear ranges of 10 ng/mL to 2.0 µg/mL and 2.0 µg/mL to 50 μg/mL with a low limit of detections (LLOD)) of 10 ng/mL and 2.0 µg/mL, respectively. The obtained results are demonstrated the accuracy of the designed nanoprobe which is as high as to be used for the MTX detection in the routine therapeutic drug monitoring. In addition, the established nanoprobe shows significant specificity towards MTX compared to the co-administered drugs. Other advantages of this nanoprobe are its simple approach, high selectivity and sensitivity for MTX determination in plasma media which can approve the applicability of this MTX nanosensor in real samples.


Molecular structures of unbound and transcribing RNA polymerase III.
Transcription of genes encoding small structured RNAs such as transfer RNAs, spliceosomal U6 small nuclear RNA and ribosomal 5S RNA is carried out by RNA polymerase III (Pol III), the largest yet structurally least characterized eukaryotic RNA polymerase. Here we present the cryo-electron microscopy structures of the Saccharomyces cerevisiae Pol III elongating complex at 3.9 Å resolution and the apo Pol III enzyme in two different conformations at 4.6 and 4.7 Å resolution, respectively, which allow the building of a 17-subunit atomic model of Pol III. The reconstructions reveal the precise orientation of the C82-C34-C31 heterotrimer in close proximity to the stalk. The C53-C37 heterodimer positions residues involved in transcription termination close to the non-template DNA strand. In the apo Pol III structures, the stalk adopts different orientations coupled with closed and open conformations of the clamp. Our results provide novel insights into Pol III-specific transcription and the adaptation of Pol III towards its small transcriptional targets.


Effect of influenza vaccination in solid organ transplant recipients: A nationwide population-based cohort study.
Vaccination can prevent influenza in solid organ transplant (SOT) recipients. Using a modified season-specific approach over nine consecutive influenza seasons, we investigated influenza vaccination coverage and effectiveness in a population-based nationwide cohort study that included all SOT recipients aged ≥18 years who were living in Denmark from December 1, 2007 to April 1, 2016. The primary outcome was the season-specific risk of all-cause pneumonia admission. Secondary outcomes were season-specific influenza-related admission, intensive care unit (ICU) admission, and all-cause mortality. Crude and adjusted hazard ratios (aHRs) with 95% confidence intervals (CIs) were estimated using Cox proportional hazards regression models. In total, 11 381 person-years of follow-up data were collected from 5745 SOT recipients, 48% of whom were vaccinated. Influenza vaccination was associated with a reduced risk of all-cause pneumonia admission (aHR, 0.83; 95% CI, 0.69-0.99; p = .035) and all-cause mortality (aHR, 0.60; 95% CI, 0.47-0.76; p = .001), but not influenza-related admission (aHR, 0.75; 95% CI, 0.46-1.22; p = .24) or ICU admission (aHR, 0.84; 95% CI, 0.67-1.06; p = .14) during the same season. Despite these benefits, uptake of influenza vaccination among SOT recipients was low. Therefore, annual influenza vaccination needs to be prioritized.


Legibility of Chinese characters in peripheral vision and the top-down influences on crowding.
Written Chinese is distinct from alphabetic languages because of its enormous number of characters with a great range of spatial complexities (stroke numbers). In this study we investigated the impact of spatial complexity on legibility of Chinese characters as well as associated crowding in peripheral vision. Our results showed that for isolated characters, threshold sizes of complex characters increased faster with retinal eccentricity than did those of simple characters, suggesting possible "within-character" crowding among parts of complex Chinese characters. However, such "within-character" crowding was rendered negligible by strong "between-character" crowding introduced by flankers. When the target and flankers belonged to different complexity groups, the intensity and extent of crowding were greatly reduced, which could be explained by top-down influences as well as lower-level mechanisms. We suggest that crowding can be attributed to multiple mechanisms at different levels of visual processing.


GABA(B) receptor-mediated ERK1/2 phosphorylation via a direct interaction with Ca(V)1.3 channels.
Neuronal L-type Ca(2+) channels play pivotal roles in regulating gene expression, cell survival, and synaptic plasticity. The Ca(V)1.2 and Ca(V)1.3 channels are 2 main subtypes of neuronal L-type Ca(2+) channels. However, the specific roles of Ca(V)1.2 and Ca(V)1.3 in L-type Ca(2+) channel-mediated neuronal responses and their cellular mechanisms are poorly elucidated. On the basis of our previous study demonstrating a physical interaction between the Ca(V)1.3 channel and GABA(B) receptor (GABA(B)R), we further examined the involvement of Ca(V)1.2 and Ca(V)1.3 in the GABA(B)R-mediated activation of ERK(1/2), a kinase involved in both CREB activation and synaptic plasticity. After confirming the involvement of L-type Ca(2+) channels in baclofen-induced ERK(1/2) phosphorylation, we examined a specific role of Ca(V)1.2 and Ca(V)1.3 channels in the baclofen effect. Using siRNA-mediated silencing of Ca(V)1.2 or Ca(V)1.3 messenger, we determined the relevance of each channel subtype to baclofen-induced ERK(1/2) phosphorylation in a mouse hippocampal cell line (HT-22) and primary cultured rat neurons. In the detailed characterization of each subtype using HEK293 cells transfected with Ca(V)1.2 or Ca(V)1.3, we found that GABA(B)R can increase ERK(1/2) phosphorylation and Ca(V)1.3 channel activity through direct interaction with Ca(V)1.3 channels. These results suggest a functional interaction between Ca(V)1.3 and GABA(B)R and important implications of Ca(V)1.3/GABA(B)R clusters for translating synaptic activity into gene expression alterations.


Dipyridyl amines: potent metabotropic glutamate subtype 5 receptor antagonists.
Modulation of the metabotropic glutamate subtype 5 (mGlu5) receptor may be useful in the treatment of a variety of central nervous system disorders. Herein, we report on the discovery, synthesis, and biological evaluation of dipyridyl amines as small molecule mGlu5 antagonists.


Career development of women in health care administration: a preliminary consideration.
As greater numbers of women enter health care administration, it is important to understand their career perceptions so that organizations can assist them in becoming effective leaders. This article reports findings from focused interviews of women administrators regarding their career development and expectations and suggests issues that require attention.


[Prophylaxis of wound infection in patients with head and neck tumors with an account of the pathogen taxonomic pattern].
The most important problem of onkology, i. e. antibiotic prophylaxis and treatment of postoperative infectious complications is touched upon in the paper. The current publications on the problem are discussed and the autors' experience on the treatment is described. The spectrum of the main pathogens of wound infections in patients with head and neck tumors is considered.


Randomized clinical trial of laparoendoscopic single-site versus conventional laparoscopic cholecystectomy.
Conventional laparoscopy with three or more ports remains the 'gold standard' for cholecystectomy, but a laparoendoscopic single-site (LESS) approach is emerging, designed to decrease parietal trauma and improve cosmesis. This study compared conventional laparoscopic (CL) with LESS cholecystectomy, with short-term clinical results as the main outcomes. A randomized trial of CL and LESS cholecystectomies involving 150 patients was undertaken. Follow-up was for 1 month after surgery. The primary endpoint was body image results evaluated by means of validated scales. Secondary endpoints were: postoperative pain measured on a visual analogue scale, analgesia requirement, morbidity, quality of life (QoL) measured with Short Form 12, duration of operation, hospital stay, time to return to work and cost analysis. Operating times and complications were similar in the two groups. Two LESS procedures (3 per cent) were converted to two-port laparoscopy owing to difficulties with exposure, and one CL operation was achieved through a single port because extensive fibrous peritoneal adhesions prevented placement of other ports. There were three and four port-site seroma/haematomas in the LESS and CL groups respectively. Better pain profiles and lower analgesia requirements were recorded in the LESS group (P < 0·001). QoL, body image and scar scale results were also better (P < 0·001). Operative costs were higher for LESS procedures (P < 0·001), although median time to return to work was shorter (P = 0·003). LESS is an alternative to CL cholecystectomy associated with better cosmesis, body image, QoL and an improved postoperative pain profile.


Substituted-letter and transposed-letter effects in a masked priming paradigm with French developing readers and dyslexics.
The aim of the study was to undertake a behavioral investigation of the development of automatic orthographic processing during reading acquisition in French. Following Castles and colleagues' 2007 study (Journal of Experimental Child Psychology, 97, 165-182) and their lexical tuning hypothesis framework, substituted-letter and transposed-letter primes were used in a masked priming paradigm with third graders, fifth graders, adults, and phonological dyslexics matched on reading level with the third graders. No priming effect was found in third graders. In adults, only a transposed-letter priming effect was found; there was no substituted-letter priming effect. Finally, fifth graders and dyslexics showed both substituted-letter and transposed-letter priming effects. Priming effects between the two groups were of the same magnitude after response time (RT) z-score transformation. Taken together, our results show that the pattern of priming effects found by Castles and colleagues in English normal readers emerges later in French normal readers. In other words, language orthographies seem to constrain the tuning of the orthographic system, with an opaque orthography producing faster tuning of orthographic processing than more transparent orthographies because of the high level of reliance on phonological decoding while learning to read.


Dopamine D2 receptor occupancy with risperidone or olanzapine during maintenance treatment of schizophrenia: a cross-sectional study.
In treating schizophrenia, it has been established that 65-80% occupancy of dopamine D2 receptors optimizes therapeutic efficacy while minimizing risks of extrapyramidal symptoms. However, it is unclear as to whether it is necessary to keep D2 receptor occupancy within this therapeutic window to maintain response. In this study, daily peak and trough D2 receptor occupancy levels were estimated in clinically stable patients with schizophrenia (DSM-IV) who were receiving risperidone or olanzapine. Using two collected plasma samples, plasma antipsychotic concentrations at peak and trough were estimated with population pharmacokinetic techniques. Corresponding dopamine D2 receptor occupancy levels were then estimated, using a recently developed model. 35 subjects with stable schizophrenia completed the study (mean±SD age, 48.8±13.8years; male [N=14]; Asians [N=23], Caucasians [N=12]; risperidone [N=20] at 3.2±2.3mg/day, and olanzapine [N=15] at 9.2±4.9mg/day) between September and December 2010. 48.6% (N=17) did not achieve a continuous blockade of ≥65%. Moreover, 11.4% (N=4) did not achieve the 65% threshold at estimated peak concentrations. In conclusion, approximately half the subjects with stable schizophrenia did not achieve estimated continuous blockade of D2 receptor occupancy of ≥65%. The results suggest that sustained D2 receptor occupancy levels of ≥65% may not always be necessary for the maintenance treatment of schizophrenia.


The crystal and molecular structure of 9-alpha-D-arabinofuranosyladenine.
The glycosyl torsional angles in two crystallographically-independent molecules of alpha-araA are -73 and -64 degrees, both of which are in the "anti" region. The sugar conformations are C(3')-endo and C(2')-exo-C(3')-endo.


Predicting congenital rubella syndrome in Japan, 2018-2019.
A rubella epidemic has been ongoing in Japan since August 2018. In the present study, we aimed to predict the likely size of a congenital rubella syndrome (CRS) epidemic during 2018-19. The expected number of CRS cases was estimated using an integral equation based on age-specific incidence of rubella among adult women, the time delay from gestational age of infection to diagnosis of CRS, and distribution of the mothers' age at delivery. We used epidemic data during 2012-14 to parameterize the model and applied this in the prediction for 2018-19. In analyzing the 2012-14 epidemic data, the mean delay from the mother's infection to diagnosis was estimated at 24.2weeks (95% confidence interval (CI): 20.7, 28.1). Applying the parameterized model, together with the more than 480 rubella cases in women in 2018 as well as delayed mother's age at delivery in 2017, we determined that the expected number of CRS cases would be 9.7 (95% CI: 6.5, 12.5) cases. As the epidemic is ongoing, the cumulative number of CRS cases could potentially reach 96.8 (95% CI: 65.3, 125.5) cases, if rubella cases in adult women rose to 10 times the number by week 49 in 2018. CRS is expected to occur an average of 24weeks following the mother's infection with rubella virus. Accounting for an increase to 650 cases in women by week 5 in 2019, the expected number of CRS cases during 2018-19 has already exceeded 13 cases, as of week 5 in 2019.


Do the stent blind insertion into the main portal vein (MPV) and stent diameter influence the surgical outcome of the transjugular intrahepatic portosystemic shunt (TIPS)?
Transjugular intrahepatic portosystemic shunt (TIPS) surgery is a clinical intervention to treat portal hypertension (PH) by deploying a covered stent to establish a shunt path for the portal vein (PV) system, and proper surgical strategy is of great importance to balance the shunt effect and the risk of complications. To understand the clinical strategies of the stent blind insertion and stent selection in clinic, this study investigated the effects of varying stent insertion positions and diameters on the PV hemodynamics and the shunt effect by computational fluid dynamics (CFD) analysis of five post-TIPS subjects. The results showed that the successful TIPS surgeries of the five PH subjects were confirmed by quantifying their pressure drops. The stent insertion positions at the main portal vein (MPV) slightly affected the clinically concerned hemodynamic indexes (i.e., MPV pressure, stent-outlet velocity) and the shunt index (SI). This indicated that the position of the stent going into the MPV may not need to be deliberately selected. Moreover, the covered stents with 6 mm and 8 mm diameters slightly influenced the hemodynamics as well, but the large-diameter stent better improved the shunt effect compared to the small-diameter one. Despite this, the 6 mm stent was suggested thanks to the higher risk of the hepatic encephalopathy (HE) observed in clinic, which indicated the excessive shunt of the 8 mm stent. The current work revealed the effects of different TIPS strategies on the surgical outcome, and could be useful for potential clinical practices.


Interleukin-1 up-regulates transcription of its own receptor in a human fibroblast cell line TIG-1: role of endogenous PGE2 and cAMP.
The regulation of interleukin-1 receptor (IL-1R) mRNA expression by IL-1 in a human lung fibroblast cell line (TIG-1) was investigated. After 2 h of stimulation with human recombinant IL-1 alpha or IL-1 beta, the levels of T cell/fibroblast-type IL-1R mRNA increased, and the elevation was sustained for at least 72 h. IL-1 also stimulated synthesis of prostaglandin E2 (PGE2) and secondary cAMP accumulation. Exogenously added PGE2 increased the levels of both IL-1R mRNA and intracellular cAMP. Forskolin, cholera toxin and 8-Bromo adenosine (8-Br-cAMP) all increased IL-1R mRNA levels. Indomethacin blocked IL-1 stimulation of IL-1R mRNA expression, PGE2 production and cAMP. 125I-labeled IL-1 alpha-binding studies showed that this cell line expresses 2.6 x 10(4) IL-1R per cell with a Kd of 5.1 x 10(-10) M. After treatment of the cells with IL-1, the level of IL-1R increased over that of control cells. PGE2 also increased IL-1R without alteration in its affinity. Cross-linking experiments indicate that this cell line expresses the 80-kDa receptor molecule before and after treatment with PGE2; the molecular mass corresponds to the T cell/fibroblast type I IL-1R. These results indicate that IL-1 does not directly stimulate expression of IL-1R mRNA or cell surface IL-1R, but only indirectly by stimulation of endogenous PGE2.


The effect of watching videos using virtual reality during operating room transfer on the fear andanxiety of children aged 6-12 undergoing inguinal hernia surgery: A randomized controlled trial.
Being in a hospital and undergoing a surgical intervention are very stressful for the children and their parents. It is critical to keep the pain, fear and anxiety of children at a minimum level during this period in order to have a better process. The aim of this study was to evaluate the effect of watching videos using virtual reality on fear and anxiety of children aged 6-12 years old during operating room transfer for inguinal hernia surgery. This prospective and randomized controlled study used parallel trial design guided by the CONSORT checklist. The study was conducted with two groups: the group that watch videos using virtual reality group and the control group. Data were collected using follow-up forms; the Children's State Anxiety Scale and the Children's Fear Scale. Children in the virtual realty group had lower anxiety and fear scores than the children in the control group (p < 0.05). Evaluation of the in-group measurements demonstrated that the fear and anxiety scores of the children in the virtual reality group decreased after the transfer compared to that of the pre-transfer, while the children in the control group experienced higher fear and anxiety scores (p < 0.05). Virtual reality is an effective method to reduce fear and anxiety in children aged 6-12 years old during their transfer to the operating room. Evidence-based protocol and guidelines should be developed for nonpharmacological methods such as virtual reality for procedural fear and anxiety in children.


Neurones in the spinal trigeminal nucleus of the cat responding to movement of the vibrissae.
1. In chloralose anaesthetised cats the receptive field organisation and response properties of neurones of the spinal trigeminal nucleus were exmained and compared with the discharge characteristics of afferent units from vibrissae. 2. The response properties of the primary afferent discharges were preserved in at least a proportion of the second order cells, 82% of those which responded to movements of the maxillary vibrissae had phasic discharges and 18% had tonic discharges. 3. The discharge characteristics of the two main types of primary afferent slowly-adapting units (St I and St II) were distinguishable in the tonically active cells of the nucleus. These two groups showed (i) substantially different interspike interval distributions, (ii) different adaptive properties and (iii) different directional sesitivity. The two categories were designated nuclear type I and type II in accordance with the classification of primary afferent slowly-adapting units. 4. A loss of stimulus information was deduced from a consistent increase in the variability of the second order discharges compared with their presumed afferent input, but this information loss may be important in allowing the linear summation of discharges at the level of the thalamus.


Can pulmonary venous hypertension be graded by the chest radiograph?
The chest radiograph is widely used for the diagnosis and follow-up of patients with pulmonary venous hypertension (PVH) caused by rheumatic heart disease. Observer accuracy and interobserver agreement on the radiographic diagnosis of PVH is rarely assessed. We have studied this accuracy and agreement in 120 consecutive patients. Patients were selected on the basis of mean pulmonary capillary wedge pressures (PCWP) obtained in the basal state. A chest radiograph was obtained within the 24 h before catheterization and was assessed by two observers blinded to the diagnosis and PCWP. PVH was graded as absent (PCWP < 12 mmHg); mild (PCWP 12-19 mmHg; redistribution of blood flow and hilar haze); moderate (PCWP 20-25 mmHg; septal oedema, pleural effusion) and severe (PCWP > 25 mmHg; alveolar oedema). Observer and interobserver agreement was quantified by using the Kappa (kappa) statistic. The radiological assessment coincided with the measured PCWP in 52% of the patients according to observer 1 and 43% of the patients according to observer 2 (kappa 0.36 and 0.24), indicating poor agreement with haemodynamic values. Interobserver agreement was poor (kappa 0.3-0.35). Observer accuracy and interobserver agreement increased when the radiographic criteria were used to distinguish patients with normal PCWP from those with abnormal pressures when 85% and 78% of radiographs were correctly classified (kappa 0.6 and 0.5) by observers 1 and 2, respectively. Overall agreement was significant (kappa 0.6). In differentiating patients with a PCWP > 20 mmHg from those with a PCWP < 20 mmHg, 78% and 75% of radiographs were correctly classified, giving kappa values of 0.6 and 0.5. Overall observer agreement was significant (kappa 0.6). The chest radiograph is useful in detecting the presence of PVH and in distinguishing significant elevations in PCWP (> 20 mmHg) from mildly elevated PCWP. There is poor correlation between haemodynamic and radiographic findings when more precise grading of the severity of PVH is attempted.


Single-molecule characterization of compressed RecA nucleoprotein filaments.
RecA is a central enzyme of homologous recombination in bacteria, which plays a major role in DNA repair, natural transformation and SOS-response activation. RecA forms nucleoprotein filaments on single-stranded DNA with a highly conserved architecture that is also shared by eukaryotic recombinases. One of the key features of these filaments is the ability to switch between stretched and compressed conformations in response to ATP binding and hydrolysis. However, the functional role of such conformational changes is not fully understood. Structural data revealed that in the absence of ATP RecA binds DNA with the stoichiometry of 5 nucleotides per one monomer, while in the presence of ATP the binding stoichiometry is 3:1. Such differences suggest incompatibility of the active and inactive conformations, yet dynamic single-molecule studies demonstrated that ATP and apo conformations can be directly interconvertible. In the present work we use a single-molecule approach to address the features of inactive RecA nucleoprotein filaments formed de novo in the absence of nucleotide cofactors. We show that compressed RecA-DNA filaments can exist with both 5:1 and 3:1 binding stoichiometry which is determined by conditions of the filament assembly. However, only a 3:1 stoichiometry allows direct interconvertibility with the active ATP-bound conformation.


Hierarchical periodic micro/nano-structures on nitinol and their influence on oriented endothelialization and anti-thrombosis.
The applications of hierarchical micro/nano-structures, which possess properties of two-scale roughness, have been studied in various fields. In this study, hierarchical periodic micro/nano-structures were fabricated on nitinol, an equiatomic Ni-Ti alloy, using a femtosecond laser for the surface modification of intravascular stents. By controlling the laser fluence, two types of surfaces were developed: periodic nano- and micro/nano-structures. Evaluation of water contact angles indicated that the nano-surface was hydrophilic and the micro/nano-surface was hydrophobic. Endothelial cells aligned along the nano-structures on both surfaces, whereas platelets failed to adhere to the micro/nano-surface. Decorrelation between the responses of the two cell types and the results of water contact angle analysis were a result of the pinning effect. This is the first study to show the applicability of hierarchical periodic micro/nano-structures for surface modification of nitinol.


[What should the paediatrician know about hyperphenylalaninaemia?].
Hyperphenylalaninaemias are defined by a blood phenylalanine over 2mg/dl. The main cause is due to a mutation in the gene that codes the phenylalanine hydroxylase that catalyses the reaction that converts phenylalanine into tyrosine. The hyperphenylalaninaemias are classified into benign or mild hyperphenylalaninaemias, or mild, moderate or classic phenylketonurias. Due to its delayed detection outside the neonatal period it causes severe mental retardation. Its detection along with congenital hypothyroidism has been part of the National Neonatal Screening Program since 1992 in Chile. This article aims to answer the most common questions asked by the paediatrician when faced with a patient with hyperphenylalaninaemias.


Hydrolysis of fungal and plant cell walls by enzymatic complexes from cultures of Fusarium isolates with different aggressiveness to rye (Secale cereale).
The efficiency of hydrolysis of fungal (Fusarium spp.) cell wall and rye root cell wall by crude enzymatic complexes from (42-day-old) cultures of three F. culmorum isolates, a plant growth-promoting rhizosphere isolate (PGPF) DEMFc2, a deleterious rhizosphere isolate (DRMO) DEMFc5, and a pathogenic isolate DEMFc37, as well as two other, pathogenic isolates belonging to F. oxysporum and F. graminearum species was studied. In the enzymatic complexes originating from the Fusarium spp. cultures, the activities of the following cell wall-degrading enzymes were identified: glucanases, chitinases, xylanases, endocellulases, exocellulases, pectinases, and polygalacturonases. The preparation originating from a culture of the PGPF isolate was the least efficient in plant cell wall (PCW) hydrolysis. There were no significant differences in the efficiency of PCW hydrolysis between preparations from cultures of the DRMO and the pathogenic isolates. PGPF was the most efficient in liberating reducing sugars and N-acetylglucosamine (GlcNAc) from fungal cell walls (FCW). Xylanase activities of the enzymatic complexes were strongly positively (R > +0.9) correlated with their efficiency in hydrolyzing PCW, whereas chitinase activities were correlated with the efficiency in FCW hydrolysis.


Perceptual complexity of lexical, surface structure, and deep structure types of ambiguous sentences and change in heart rate.
Studies comparing lexical, surface-, and deep-structure types of ambiguous sentences with unambiguous ones for perceptual complexity show conflicting findings mainly because of task artifacts and lack of adequate control of confounding variables. In the present study, three types of ambiguous sentences and matched unambiguous controls were compared. Acceleration and deceleration measures of heart rate (HR) were used since these have been shown to reliably indicate complexity of cognitive activity. The study used a Groups x Ambiguity Condition x Ambiguous Sentence-Type x Sentence-Clusters repeated measures Latin Square ANOVA design which permitted isolation of variance related to the specific sample of sentences used. Eighteen low-bias ambiguous sentences (six from each type) and their control pairs, divided into three lists, were presented to 30 male undergraduates. Analysis of data showed: (1) significant Ambiguity x Type and Ambiguity effects for the percentage of increase in HR during processing of the sentence and, (2) significant effect of Ambiguity for the percentage of decrease in HR. These results were interpreted as showing that while low-bias ambiguous sentences are perceptually more complex than unambiguous ones, the effect of ambiguity may be greater for deep-structure type of ambiguous sentences than for lexical and surface-structure types.


Factors contributing to deletion within Mungbean yellow mosaic virus partial dimers in binary vectors used for agroinoculation.
Mungbean yellow mosaic virus-Vigna (MYMV) sequences cloned as partial dimers within the T-DNA of a binary vector were deleted at a high frequency upon conjugal mobilization from Escherichia coli into Agrobacterium tumefaciens. This deletion involving the genome-length viral DNA did not occur when the binary plasmid was inside E. coli and when the binary plasmid was introduced into Agrobacterium by electroporation. Deletions occurred in both DNA A and DNA B partial dimers. A minimum of 500-nt continuity on either side of the nonanucleotide in the duplicated common region is required for deletion. A. tumefaciens cells in which deletion was complete, grew as larger colonies reflecting a growth advantage. The small, slow-growing colonies eventually lost the genome-length viral sequences after a few more cycles of growth. Partial dimers in binary plasmids pGA472 and pBin19 with RK2 replicon underwent deletion while those in pPZP with pVS1 replicon did not undergo deletion. Deletion was observed in A. tumefaciens strains C58, A136, A348 and A281 with C58 chromosome background, but not in Ach5 and T37. Interestingly, deletion did not occur in A. tumefaciens strain AGL1 with a recA mutation in C58 chromosome, implying a clear role for recombination in deletion. These observations suggest the choice of Agrobacterium strains and binary vectors for agroinoculation of geminiviruses.


Swallowing function and tracheotomy dependence after combined-modality treatment including free tissue transfer for advanced-stage oropharyngeal cancer.
There are many treatments available for advanced oropharyngeal cancer. Organ-sparing protocols reserve surgery for salvage and are thought to provide adequate rehabilitation. Surgical resection with free tissue transfer may also provide adequate functional rehabilitation. The objective was to describe swallowing status and time to decannulation in a series of patients treated with combined-modality therapy that included free flap reconstruction. Retrospective chart review. Patient data were obtained from medical records of 20 patients with stage III or IV oropharyngeal carcinoma, who were consecutively treated with surgical tumor extirpation, free flap reconstruction, and postoperative irradiation at a tertiary academic center from 1985 to 2002. The following variables were identified: patient and tumor characteristics, free flap type, irradiation data, and airway and swallowing status before and after treatment. One patient underwent total laryngopharyngectomy, and the remaining 19 patients underwent tracheotomy at the time of definitive surgery. Free flap reconstructions included 1 ulnar and 15 radial forearm fasciocutaneous flaps and 4 fibula osteocutaneous flaps. Postoperatively, all 19 tracheotomized patients had successful decannulation. Average time to decannulation was 15 days (range, 3-42 d). After surgery and before irradiation, 13 patients initiated oral intake, on average, at 19.5 days (range, 7-28 d); 6 patients required no additional supplementation. By 4 months after surgery, having completed radiation therapy, 10 patients were consuming all nutrition orally; the other 10 patients still required tube-feed supplementation, although 6 of these patients were also eating by mouth. Combined-modality treatment that includes free flap reconstruction for advanced-stage oropharyngeal cancer may provide reasonable functional rehabilitation with respect to postoperative airway and swallowing.


Expression of the Hepatitis C Virus core-NS3 Fusion Protein on the Surface of Bacterial Ghosts: Prospects for Vaccine Production.
Antigen presentation using bacterial surface display systems, on one hand, has the benefits of bacterial carriers, including low-cost production and ease of manipulation. On the other hand, the bacteria can help in stimulating the immune system as an adjuvant. For example, using bacterial surface display technology, we developed a hepatitis C virus (HCV) multiple antigens displaying bacteria's surface and then turned it into a bacterial ghost. The HCV core and NS3 proteins' conserved epitopes were cloned into the AIDA gene plasmid as an auto transporter. The recombinant plasmid was then transformed into Escherichia coli (E. coli) Bl21 (DE3). Recombinant bacteria were then turned into a bacterial ghost, an empty cell envelope. Whole-cell ELISA, flow cytometry, and Western blot techniques were used for monitoring the expression of proteins on the surface of bacteria. A fusion protein of HCV core-NS3-AIDA was successfully expressed on the E. coli Bl21 (DE3) surface and confirmed by western blotting, Enzyme-Linked Immunosorbent Assay (ELISA), and flow cytometry detection techniques. The presence of HCV antigens on non-pathogen bacteria surfaces holds promise for developing safe and cost-benefit-accessible vaccines with optimal intrinsic adjuvant effects and exposure of heterologous antigens to the immune system.


National hepatitis registry in Pakistan: a dire need for hepatitis surveillance and control.
Hepatitis is a major public health issue in Pakistan, with an estimated 11.55% prevalence of HCV infection in the adult population. The country ranks second globally in terms of hepatitis C virus (HCV) infections, with approximately one in every 20 Pakistanis already infected. The mortality rates due to HBV and HCV stand at 563,000 and 366,000 annually, respectively. However, the absence of a national registry or database system and the lack of coordination among provinces pose significant obstacles in combating this disease effectively. To address this issue, the establishment of a centralized national database registry is crucial, allowing comprehensive analysis, tracking of hepatitis prevalence, and identification of high-risk areas for targeted interventions. By fostering collaboration among provinces, the government, and non-governmental organizations, the registry would facilitate joint decision-making, minimize duplication of efforts, and address inconsistencies in diagnosis and treatment. Collaborating with student-run organizations and leveraging enhanced laboratory capacities post-COVID era can strengthen the hepatitis control program. The centralized approach and unified efforts are necessary to achieve the goal of a hepatitis-free Pakistan, where a healthier future can be realized.


Stereotactic Radiation Therapy versus Brachytherapy: Relative Strengths of Two Highly Efficient Options for the Treatment of Localized Prostate Cancer.
Stereotactic body radiation therapy (SBRT) has become a valid option for the treatment of low- and intermediate-risk prostate cancer. In randomized trials, it was found not inferior to conventionally fractionated external beam radiation therapy (EBRT). It also compares favorably to brachytherapy (BT) even if level 1 evidence is lacking. However, BT remains a strong competitor, especially for young patients, as series with 10-15 years of median follow-up have proven its efficacy over time. SBRT will thus have to confirm its effectiveness over the long-term as well. SBRT has the advantage over BT of less acute urinary toxicity and, more hypothetically, less sexual impairment. Data are limited regarding SBRT for high-risk disease while BT, as a boost after EBRT, has demonstrated superiority against EBRT alone in randomized trials. However, patients should be informed of significant urinary toxicity. SBRT is under investigation in strategies of treatment intensification such as combination of EBRT plus SBRT boost or focal dose escalation to the tumor site within the prostate. Our goal was to examine respective levels of evidence of SBRT and BT for the treatment of localized prostate cancer in terms of oncologic outcomes, toxicity and quality of life, and to discuss strategies of treatment intensification.


Plasma Fibrinogen Is a Natural Deterrent to Amyloid Beta-Induced Platelet Activation.
Alzheimer's disease (AD) is a devastating neurodegenerative disorder, characterized by extensive loss of neurons, and deposition of amyloid beta (Aβ) in the form of extracellular plaques. Aβ is considered to have critical role in synaptic loss and neuronal death underlying cognitive decline. Platelets contribute to 95% of circulating amyloid-precursor protein that releases Aβ into circulation. We have recently demonstrated that, Aβ active fragment containing amino acid sequence 25-35 (Aβ25-35) is highly thrombogenic in nature, and elicits strong aggregation of washed human platelets in RhoA-dependent manner. In the present study we evaluated the influence of fibrinogen on Aβ-induced platelet activation. Intriguingly, Aβ failed to induce aggregation of platelets suspended in plasma but not in buffer. Fibrinogen brought about dose-dependent decline in aggregatory response of washed human platelets elicited by Aβ25-35, which could be reversed by increasing doses of Aβ. Fibrinogen also attenuated Aβ-induced platelet responses like secretion, clot retraction, rise in cytosolic Ca+2 and reactive oxygen species (ROS). Fibrinogen prevented intracellular accumulation of full length amyloid beta peptide (Aβ42) in platelets as well as neuronal cells. We conclude that fibrinogen serves as a physiological check against the adverse effects of Aβ by preventing its interaction with cells.


Re-evaluating the predictive roles of metabolic complications and clinical outcome according to eGFR levels--a four-years prospective cohort study in Taiwan.
Metabolic complications are associated with clinical outcomes in patients with chronic kidney disease (CKD). These outcomes differ among patients according to the different stages of disease. The prevalence and association of type and number of metabolic complications with renal progression and death in patients having different eGFR levels has high clinical value, but this fact has been rarely evaluated in prospective studies. We prospectively followed a cohort of 1157 CKD patients from 2006 to death or until 2010, and evaluated the prevalence of CKD-related complications and their association with renal progression (defined as a decline in eGFR by > 50% from baseline, or end-stage renal disease requiring dialysis) and death in patients with eGFRs above and below 45 mL/min/1.73 m(2) using Cox-proportional hazard models. The estimated rate (per 100 patient-years) of renal progression and death were 11.9 and 4.9, respectively. The eGFR thresholds determined by ROC analysis with a sensitivity of 90% for any metabolic complication were 60.8 mL/min/1.73 m(2) and 74.3 mL/min/1.73 m(2) using the MDRD and CKD Epidemiology Collaboration equations, respectively. CKD-related complications associated with renal progression in patients having eGFR < 45 mL/min/1.73 m(2) were hyperphosphatemia, anemia, microinflammation and hypoalbuminemia. Those CKD-related complications associated with death were hypoalbuminemia and hyperuricemia. Hypoalbuminemia predicted renal progression, and, hypoalbuminemia and microinflammation predicted death in patients with eGFR ≥ 45 mL/min/1.73 m(2). The number of complications (≥ 3) independently predicted both endpoints in patients with eGFR < 45 mL/min/1.73 m(2). Hypoalbuminemia was a unique and strong predictor of renal progression and all-cause mortality in CKD patients, independent of their demographic characteristics, traditional risk factors, renal function severity, the presence of cardiovascular disease and other metabolic abnormalities. Most other metabolic complications and the number of complications (≥ 3) were associated with the clinical outcomes of patients with eGFR < 45 mL/min/1.73 m(2) rather than in those with higher eGFRs. The findings from the present study offer a novel insight into the association between metabolic complications and patient outcomes and may help to refine risk stratification according to disease stage.