Effective transduction and stable transgene expression in human blood cells by a third-generation lentiviral vector.
Difficulty in gene transduction of human blood cells, including hematopoietic stem cells, has hampered the development of gene therapy applications for hematological disorders, encouraging the development and use of new gene delivery systems. In this study, we used a third-generation self-inactivating (SIN) lentiviral vector system based on human immunodeficiency virus type 1 (HIV-1) to improve transduction efficiency and prevent vector-related toxicity. The transduction efficiency of the HIV-1-based vector was compared directly with the Moloney murine leukemia virus (MLV) SIN vector in human leukemia cell lines. Initial transduction efficiencies were almost 100% for the HIV and less than 50% for the MLV vectors. Similar results were observed in 11 types of primary cells obtained from leukemia or myeloma patients. Transgene expression persisted for 8 weeks in cells transduced with the HIV vector, but declined with the MLV vector. In addition, resting peripheral blood lymphocytes and CD34(+) hematopoietic cells were transduced successfully with the HIV vector, but not with the MLV vector. Finally, we confirmed vector gene integration in almost all colony-forming cells transduced with the HIV vector, but not with the MLV vector. In conclusion, this lentiviral vector is an excellent gene transduction system for human blood cells because of its high gene transduction and host chromosome integration efficiency.


Large asymptomatic carcinoid tumor in an HIV-infected man.
Many different tumor types have been described in patients with AIDS; carcinoid tumors, however, are a distinct exception to this phenomenon. Our case illustrates the asymptomatic presentation of a large abdominal carcinoid in an HIV-infected man, and further expands the list of neoplasms that have been found in this population.


The R362A mutation at the C-terminus of CA inhibits packaging of human immunodeficiency virus type 1 RNA.
The capsid (CA) sequence of human immunodeficiency virus type 1 (HIV-1) Gag protein consists of two independently folded domains named the N-terminal domain (NTD) and C-terminal domain (CTD) that are connected by a flexible linker. Most of the CTD sequence adopts rigid structure except for the last 11 amino acids (positions 354 to 364) that are disordered even in the context of the downstream SP1 and nucleocapsid (NC) sequence. Although disordered, this short peptide region plays a crucial role in HIV-1 replication. In this study, we identified three second-site mutations within Gag named A238T, G358S, and N373K that rescued a deleterious mutation R362A located at the C-terminus of CA. A238T is located within the NTD of CA, G358S and N373K are positioned proximal to R362A. One of the mechanisms underlying this compensation event is correction of reduced packaging of viral RNA into the R362A mutated viruses, as shown by the results of RNase protection assays, native Northern blots experiments as well as filter-binding assays. These data suggest that one potential function for the C-terminal disordered sequence of CA in HIV-1 replication is to regulate HIV-1 RNA packaging.


SPRINT and the Kidney: What Have We Learned?
To review the impact of the Systolic Blood Pressure Interventional Trial (SPRINT) on renal function and chronic kidney disease. Hypertension is a risk factor for cardiovascular and kidney outcomes in patients with chronic kidney disease (CKD). The benefits of intensive blood pressure lowering in the CKD population in previous studies are unclear. The SPRINT compared standard (< 140 mmHg) and intensive (< 120 mmHg) blood pressure management in nondiabetic patients with high risk of cardiovascular disease. In the subgroup of patients with CKD, the most important finding was that intensive blood pressure lowering is associated with lower risk of cardiovascular disease and mortality. Other than lower levels of albuminuria, there was no benefit on clinical kidney outcomes with the intensive treatment group. The risk of incident CKD and episodes of acute kidney injury was higher in patients in the intensive treatment group, though most patients with acute kidney injury recovered kidney function. While the benefit of intensive blood pressure lowering on cardiovascular events and mortality with intensive blood pressure lowering is clear in patients with CKD, longer term follow-up may be needed to fully understand the effect on kidney function.


Fibroblast-derived exosomal microRNA regulates NKX3-1 expression in androgen-sensitive, androgen receptor-dependent prostate cancer cells.
Androgen deprivation therapy (ADT) targeting androgen production and androgen receptor (AR) signaling is the primary antihormonal therapy in the treatment of advanced prostate cancer (PCa). However, no clinically established molecular biomarkers have been identified to predict the effectiveness of ADT before starting ADT. The tumor microenvironment of PCa contains fibroblasts that regulate PCa progression by producing multiple soluble factors. We have previously reported that AR-activating factor-secreted fibroblasts increase the responsiveness of androgen-sensitive, AR-dependent PCa cells to ADT. Thus, we hypothesized that fibroblast-derived soluble factors may affect cancer cell differentiation by regulating cancer-related gene expression in PCa cells and that the biochemical characteristics of fibroblasts may be used to predict the effectiveness of ADT. Here, we investigated the effects of normal fibroblasts (PrSC cells) and three PCa patient-derived fibroblast lines (pcPrF-M5, -M28, and -M31 cells) on the expression of cancer-related genes in androgen-sensitive, AR-dependent human PCa cells (LNCaP cells) and three sublines showing different androgen sensitivities and AR dependencies. The mRNA expression of the tumor suppressor gene NKX3-1 in LNCaP cells and E9 cells (which show low androgen sensitivity and AR dependency) was significantly increased by treatment with conditioned media from PrSC and pcPrF-M5 cells but not from pcPrF-M28 and pcPrF-M31 cells. Notably, no upregulation of NKX3-1 was observed in F10 cells (AR-V7-expressing, AR-independent cells with low androgen sensitivity) and AIDL cells (androgen-insensitive, AR-independent cells). Among 81 common fibroblast-derived exosomal microRNAs that showed 0.5-fold lower expression in pcPrF-M28 and pcPrF-M31 cells than in PrSC and pcPrF-M5 cells, miR-449c-3p and miR-3121-3p were found to target NKX3-1. In only LNCaP cells, the NKX3-1 mRNA expression was significantly increased by transfection of an miR-3121-3p mimic but not that of the miR-449c-3p mimic. Thus, fibroblast-derived exosomal miR-3121-3p may be involved in preventing the oncogenic dedifferentiation of PCa cells by targeting NKX3-1 in androgen-sensitive, AR-dependent PCa cells.


Tissue-specific granularity of gap junction cytoplasmic surfaces revealed by rapid-freeze, deep-etch replicas.
Previous rapid-freeze, deep-etch replica studies have revealed the differences between heart and liver gap junctions; cytoplasmic surfaces of in situ and phenylmethylsulfonyl fluoride (PMSF)-unproteolyzed isolated cardiac gap junctions (MW 47 kD) have a particulate substructure, which is absent both in the proteolyzed heart junctions (MW 29 kD) and in the liver junctions isolated with PMSF (MW 28 kD). The present deep-etch replica studies of gap junction cytoplasmic surface (CS) membranes in several tissues of rats and mice were performed to examine whether or not this difference between liver and heart is typical of variations in gap junction proteins from tissue to tissue. In surface mucous cells of the stomach, intestinal epithelial cells, and kidney tubule cells, these epithelial gap junctions always showed smooth cytoplasmic surfaces, similar to the liver gap junctions. In contrast, in the atrial myocardium, aortic endothelium, and the ciliary process, cytoplasmic surface membranes of the gap junctions consistently revealed particulate patterns. Close examinations disclosed that those granular structures were not merely attached to the memvrane surface, but they also protruded from the membrane interior as an integral component of gap junction particles. Furthermore, in the pregnant rat uterus at term, cytoplasmic surface membranes of myometrial smooth muscle gap junctions were particulate, but those of endometrial epithelium were smooth. The present observation suggest that tissue specificity exists in cytoplasmic surface structures of gap junctions between the "true" epithelial and the nonepithelial tissues: the nonepithelial gap junctions contain the additional cytoplasmic surface domain that is absent in the gap junctions of "true" epithelial origin.


Preoperative patient factors and anatomy do not predict who will develop reflux after per oral endoscopic myotomy.
Per oral endoscopic myotomy (POEM) has been shown to be an efficacious and safe therapy for the treatment of achalasia. Compared to laparoscopic Heller myotomy however, no antireflux procedure is routinely combined with POEM and therefore the development of symptomatic or silent reflux is of concern. This study was designed to determine if various patient factors and anatomy would predict the development of gastroesophageal reflux disease post-operatively. This was a retrospective cohort study of all patients who underwent a POEM at a single institution by a single surgeon over an eight-year period (2014-2022). It has been our practice to obtain a postoperative ambulatory pH test on all patients 6 months after POEM off all acid reducing medications. Patients without a postoperative ambulatory esophageal pH monitoring test were excluded. Age, sex, obesity (BMI > 30), achalasia type, presence of a hiatal hernia, history of prior endoscopic achalasia treatments or myotomy were analyzed using univariate analysis as predictive factors for the development of postoperative GERD (DeMeester score > 14.7 on ambulatory pH monitoring). There were 179 total patients included in the study with 42 patients (23.5%) having undergone postoperative ambulatory pH testing. The majority of patients (137 or 76.5%) were lost to follow up and did not undergo ambulatory pH testing. Twenty-three out of those 42 patients (55%) had evidence of GERD on ambulatory pH testing. Multiple preoperative patient characteristics including demographics, manometric results, EGD findings, and history of prior achalasia interventions did not correlate with the development of post-operative GERD. Despite the high rate of reflux after POEM, there does not appear to be any reliable preoperative indicators of which patients have a higher risk of developing post-operative GERD after POEM.


99MTc-HIDA, a gallbladder imaging agent: experimental aspects.
The molecule N-(2,6-dimethyl-phenyl-carbamoyl-methyl)-iminodiacetic acid (HIDA), capable of chelating reduced 99mTc, was synthesized, characterized, labeled with 99mTc, and studied in experimental animals. The results indicated that the new 99mTc-radiopharmaceutical is rapidly cleared from the blood to the liver, then rapidly removed to the gallbladder and excreted into the duodenum through the common bile duct. A comparative kinetic study of 99mTc-HIDA and 131I-Rose Bengal performed in rabbits demonstrated that both radiopharmaceuticals had a similar blood clearance rate, but cleared at a different rate from liver to gallbladder. 99mTc-HIDA showed a faster accumulation in the gallbladder than 131I-Rose Bengal. These findings, combined with the advantage of the low acute toxicity of HIDA, were promising enough to encourage a further evaluation and clinical investigation of this new Tc-99m hepatobiliary agent.


Menstruation and the menopausal transition.
This paper characterizes changes in menstrual bleeding during perimenopause,including bleeding changes that represent markers of the menopausal transition. Recent results from the Study of Women's Health Across the Nation and other cohort studies are reviewed. Emerging data describing subpopulation differences in the transition experience are highlighted. When treating women in the midlife, clinicians should pay careful attention to medical factors, including both conditions and treatments, that may increase menstrual blood loss or alter menstrual cycle characteristics sufficiently to obscure the onset of the menopausal transition or the final menstrual period.


Comprehensive Approach to Lower Blood Pressure (CALM-BP): a randomized controlled trial of a multifactorial lifestyle intervention.
Complementary medicine advocates the use of a multifactorial approach to address the varied aspects of hypertension. The aim of this study was to compare the blood pressure (BP) effect and medication use of a novel Comprehensive Approach to Lowering Measured Blood Pressure (CALM-BP), based on complementary medicine principles, with the standard recommended Dietary Approach to Stop Hypertension (DASH). A total of 113 patients treated with antihypertensive drugs were randomly assigned to either CALM-BP treatment (consisting of rice diet, walks, yoga, relaxation and stress management) or to a DASH+exercise control group (consisting of DASH and walks). Ambulatory 24-h and home BP were monitored over a 16-week programme, followed by 6 months of maintenance period. Medications were reduced if systolic BP dropped below 110 mm Hg accompanied by symptoms. In addition to BP reduction, medications were reduced because of symptomatic hypotension in 70.7% of the CALM-BP group compared with 32.7% in the DASH group, P<0.0001. After 6 months, medication status was not altered in the majority of individuals. Significant reductions in body mass index, cholesterol and improved quality-of-life scores were observed only in the CALM-BP group. Lifestyle and diet modifications based on complementary medicine principles are highly effective with respect to BP control, medication use and cardiovascular risk factors.


Divergence in pharmacokinetic parameters of quinidine obtained by specific and nonspecific assay methods.
Previously published estimates of pharmacokinetic characteristics of quinidine can be shown to be dependent on whether the investigators have used analytical methods which are specific for quinidine. Areas under the plasma concentration-time curve and peak plasma concentrations after administration of the drug were higher and clearance values consequently lower in studies utilizing nonspecific assays unable to distinguish quinidine from its metabolities. The error introduced is larger after oral administration as a result of marked first-pass metabolism of quinidine. The absolute oral bioavailabilities from pharmaceutical preparations might therefore be estimated higher in studies with assays including metabolites in the determination. Although the pharmacodynamic response to quinidine is related to the plasma concentration, the therapeutic window of drug concentrations has been defined only using nonspecific assays. In light of the availability of newly developed specific assays, redefinition of the range of therapeutic plasma concentrations is opportune.


Conservative management of a severely alloimmunized pregnancy: case report.
A pregnancy complicated by severe alloimmunization because of anti-E antibodies was evaluated with fetal movement counts, nonstress tests, and serial ultrasonography-amniocentesis. The successful outcome of this case suggests that such treatment may be an acceptable alternative to intrauterine transfusions for very select cases of severe alloimmunization.


Laccaria murina and L. pumila from Pakistan, a study based on SEM and nrITS molecular marker.
Two species of the genus Laccaria, are described as new reports for Pakistan. Laccaria murina has been collected from a Himalayan moist temperate forest in Khanspur, KP, while L. pumila was found in the moist temperate forests of Kumrat Valley in Dir Upper, KP, and at higher altitudes of the Deosai plains of Gilgit-Baltistan, Pakistan. Phylogenetic analysis based on the nrITS region clustered the Pakistani species with L. murina sequences with other Asian collections and L. pumila from USA and Netherlands. SEM of basidiospores along with detailed micro-morphological data are provided. General distribution, habitat, ecology, and diagnostic features are also discussed. RESEARCH HIGHLIGHTS: It has new reports from Pakistan, studies based on light, scanning electron microscopy, and nrITS molecular markers. These species have been described with detailed micro-morphological and molecular phylogenetic analyses. General distribution, ecology, diagnostic features, and comparisons with closely related specimens have been provided. Graphical representation of DNA extraction and geographical locations of sampling sites are also illustrated (Figures 1 and 2). Very few members of this genus are already described from Pakistan.


High-level transfection of primary rabbit T lymphocytes.
Efficient gene delivery is essential for the assessment of transgene function in untransformed hematopoietic cells. Here, we explored the utility of different non-viral and viral gene delivery techniques for primary T cells from New Zealand White rabbits. We find that electroporation and nucleofection result in high-level transgene expression from both small and large GFP reporter constructs in activated rabbit T cells at moderate cytotoxicity. Both non-viral gene delivery methods were vastly superior to retroviral, lentiviral, or adenoviral transduction approaches. The effectiveness of non-viral gene delivery for functional analyses was demonstrated by downregulation of CD4 cell surface molecules through transient expression of the endocytosis-inducing Nef protein from human immunodeficiency virus in a signature motif-specific manner. This study establishes conventional electroporation as an efficient and inexpensive procedure to render primary rabbit T cells accessible to rapid functional ex vivo analyses.


Accuracy of real-time vs. blinded offline diagnosis of neoplastic colorectal polyps using probe-based confocal laser endomicroscopy: a pilot study.
Probe-based confocal laser endomicroscopy (pCLE) is a new imaging modality that enables histological examination of gastrointestinal mucosa during endoscopic procedures. Most studies have evaluated offline interpretation of pCLE images. In clinical practice, real-time interpretation is necessary to assist decision-making during the procedure. The aim of this pilot study was to compare the accuracy of real-time pCLE diagnosis made during the procedure with that of blinded offline interpretation to provide accuracy estimates that will aid the planning of future studies. pCLE was performed in patients undergoing screening and surveillance colonoscopy. Once a polyp had been identified, one endoscopist analyzed pCLE images during the procedure and made a provisional "real-time" diagnosis. Saved video recordings were de-identified, randomized, and reviewed "offline" 1 month later by the same endoscopist, who was blinded to the original diagnoses. Images from a total of 154 polyps were recorded (80 neoplastic, 74 non-neoplastic). The overall accuracy of real-time pCLE diagnosis (accuracy 79%, sensitivity 81%, specificity 76%) and offline pCLE diagnosis (83%, 88%, and 77%, respectively) for all 154 polyps were similar. Among polyps < 10 mm in size, the accuracy of real-time interpretation was significantly lower (accuracy 78%, sensitivity 71%, specificity 83%) than that of offline pCLE interpretation (81%, 86%, 78%, respectively). For polyps ≥ 10 mm, the accuracy of pCLE diagnosis in real-time was better (accuracy 85%, sensitivity 90%, specificity 75%) than offline pCLE diagnosis (81%, 97%, and 50%, respectively). These results suggest that real-time and offline interpretations of pCLE images are moderately accurate. Real-time interpretation is slightly less accurate than offline diagnosis, but overall both are comparable. Additionally, there was contrasting accuracy between the two methods for small and large polyps.


Can the Hounsfield unit predict the success of medical expulsive therapy using silodosin in 4- to 10-mm distal ureteral stones?
We aimed to investigate the predictive ability of the Hounsfield unit (HU) on non-contrast computed tomography (NCCT) for the success of medical expulsive therapy (MET) using silodosin in distal ureteric stones of 4-10 mm. The data of patients who underwent MET were retrospectively screened. The patients were divided into two groups as Groups 1 and 2 depending on the presence or absence of stone expulsion, respectively. In addition to HU calculated using the NCCT images, state of the collecting systems, daily fluid intake, number of emergency department visits, and number of pain attacks were compared. A total of 88 patients were included in the study. Sixty-four patients (72.7%) expelled the stone after MET while the treatment was not successful in 24 patients (27.3%). The stone area was significantly larger in Group 2 (28.4 ± 15.7 mm2 vs 46.8 ± 16.1 mm2 ; P < .001). NCCT-HU was calculated as 542.5 ± 256.8 for Group 1 and 873.1 ± 335.2 for Group 2, indicating a significant difference (P < .001). The mean number of pain attacks was 1.5 ± 1.2 in Group 1 and 2.2 ± 1.4 in Group 2 (P = .048). The number of visits to the emergency department significantly differed between Groups 1 and 2 (1.1 ± 1.0 and 1.8 ± 1.3, respectively; P = .010). In this study, HU and stone area values calculated on NCCT were found to be effective factors in predicting the treatment success for MET. Therefore, we consider that it would be useful to consider these parameters in the selection of an appropriate treatment for distal ureteric stones.


Good outcomes of elbow arthroscopy-assisted absorbable screw fixation for capitellum fracture of the humerus in children: a case series.
Capitellum fractures are rare coronal fractures of the distal humerus which accounts for 6% of all distal humeral fractures and only 1% of all elbow fractures. The purpose of this study was to investigate the efficacy and complications of arthroscopically assisted reduction and fixation with absorbable screws for capitellar fracture of the humerus in children. This was a retrospective case series study, which considered four patients (4 elbows), ranging from 10 to 15 years old, who were treated by arthroscopic-assisted percutaneous absorbable screw between 2018 and 2020. The elbow flexion-extension and forearm supination-pronation ranges of motion (ROM) were measured at the preoperative examination and last follow-up examination. Finally, the clinical and radiological results were assessed. The result of operations is satisfactory. The mean follow-up was 3.0 years (range 2-3.8 years). Average range of motion significantly improved from pre- to postoperation, with forearm supination from 60°(50°-60°) to 90°(90°) and pronation improved from 75°(70°-80°) to 90°(90°). The postoperative elbow flexion-extension range of motion was significantly higher compared with range of motion before surgery (P < 0.001; r = 0.949). At the final follow-up examination, the Mayo Elbow Performance Score was excellent. Satisfactory clinical results were achieved in all patients, and no postoperative complications were observed. It is an effective and safe surgical option to use arthroscopic-assisted percutaneous absorbable screw fixation for treating capitellum fracture of the humerus without any complications in children. Level IV; case series.


CD4-/CD8- double-negative tumor-infiltrating lymphocytes expanded from solid tumor tissue suppress the proliferation of tumor cells in an MHC-independent way.
Tumor-infiltrating lymphocytes (TILs) have shown remarkable clinical responses in some patients with advanced solid tumors. As a rare subset of TILs, CD4-/CD8- double-negative T cells (DNTs) were poorly known. This study aims to investigate the characteristics and function of CD3+CD4-CD8- TILs (double-negative TIL, DN-TILs) derived from solid tumor. DN-TILs were derived and expanded ex vivo from resected gastric carcinoma tissue and phenotyped by flow cytometry. The cytotoxicity of DN-TILs was determined against established tumor cell lines in vitro or through in vivo adoptive transfer into xenograft models. K562 cells were transferred with the HLA gene to verify whether the cytotoxicity of DN-TILs was MHC-independent. Flow cytometric analysis revealed a high-purity population of DN-TILs (> 97%) within CD3+ TILs, which expanded more than 800-folds in 2 weeks, consisting of a mixture of alpha-beta (αβ) and gamma-delta (γδ) T-cell receptor (TCR)-expressing cells (with the majority being αβ-TCR, > 95%). Using single-cell RNA sequencing, the expanded DN-TILs were categorized into four main subsets, Natural Killer T cells (approximately 80%, 5563 in 7028), Progenitor cells, Germ cells and T helper2 cells. DN-TILs exhibited a broad anticancer cytotoxicity in a donor-unrestricted manner against various cancer cell lines derived from pancreatic cancer (Panc-1), gastric cancer (HGC-27), ovarian cancer (SKOV-3), malignant melanoma (A375). The cytotoxicity was MHC-independent, which was not altered in K562 transferring with HLA gene or not. DN-TILs significantly reduced tumor volume in xenograft models with superior tumor-homing ability and low off-target toxicity. Gastric carcinoma derived DN-TIL can target tumor cells in vitro and in vivo. DN-TILs have the potential to be used as a adoptive cell therapy for solid cancers with both the advantages of DNT and TIL.


Development of multi-purpose 3D printed phantoms for MRI.
This work describes the development and application of 3D printed MRI phantoms. Unlike traditional phantoms these test objects are made from solid materials which can be imaged directly without filling. The models were manufactured using both MRI visible and invisible materials. The MRI visible materials were imaged on a 3T system to quantify their T 1 and T 2 properties and CT to quantify the electron density. Three phantoms are described: a distortion phantom was imaged on an open bore MRI system to assess distortion over a 30 cm field-of-view; a solid tumour model was imaged using a motion simulator and compared to a standard water phantom to assess reduction in artefacts; finally, a test object created for textural analysis was evaluated on two 3T systems and reproducibility was assessed. Material 1 was the main material used in all phantom models and has a T 1 and T 2 of 152.3 ± 3.7 ms and 56.7 ± 2.5 ms and a CT density of 127.9 HU. Material 2 had a CT density of 115.1 HU and material 3 had a T 1 and T 2 of 149.5 ± 2.9 ms and 68.8 ± 7.8 ms and CT density of 15.3 HU. Image tests demonstrated the suitability and advantage of each phantom over more traditional versions: a high density set of control points enabled a comprehensive measurement of geometric accuracy; sufficient signal with a reduction in artefact was observed in the motion phantom, and the texture model provided reproducible measurements with an ICC > 0.9 for over 76% of texture features. Three different phantoms have been successfully manufactured and used to demonstrate the application of 3D printable materials for MRI phantoms.


[Balloon catheter for cervical ripening on scarred uterus with unfavorable cervix: multicenter retrospective study of 151 patients].
To evaluate safety and efficacy of balloon catheter for labor induction in women with previous cesarean section. In a multicenter retrospective cohort study, 151 patients were included with the following criteria: pregnancy over 37 weeks, singleton, vertex presentation, previous caesarean section with unique transversal segmentary incision, medical indication for induction of labor, unfavorable cervix with Bishop score inferior to 7, no premature rupture of membranes. Balloon catheter used for cervix ripening, is inflated from 30 to 80 mL of sterile of NaCl and is left until 24 hours. Overall rate of vaginal delivery was 53.7% (81/151). Labor began before balloon catheter removal for 58 out of 151 (38.4%) with vaginal delivery for 75% (42/58). Best prognosis factors for vaginal delivery were spontaneous labor after balloon removal (P=0.004) and anterior vaginal delivery (P=0.03). Side effects were rare bleeding or PROM, but didn't prevent continuing ripening labor. Other morbidity consisted in two uterus ruptures (1.2%) without maternofetal incidence. Supracervical balloon is a safe and efficiency method for inducing labor on scarred uterus with unfavorable cervix with low side effects.


Association between self-esteem and fear of cancer recurrence in cancer survivors: A cross-sectional study.
There may be a correlation between self-esteem as an important psychological resource for individuals and Fear of cancer recurrence (FCR), but the relationship between the two is unclear. The aim of our investigation was to evaluate the association of FCR and self-esteem in cancer survivors. Cross-sectional sampling was used to select cancer survivors. The study instruments used included: General information questionnaire, Rosenberg Self-Esteem Scale, Perceived Social Support Scale, Fear of Cancer Recurrence Inventory- Shorter Form. We used logistic regression, where models were adjusted considering confounding variables to determine odds ratios (ORs) with 95% confidence intervals (CIs) for the association of FCR with self-esteem. Between February 2022 to July 2022, we screened 380 participants for eligibility, of whom 348 were included in the study. The percentage of cancer survivors who experienced clinical level of FCR was 73.9% and the self-esteem score was 27.73 ± 3.67 at a moderate level. The Pearson' s correlation coefficient indicated a significant inverse relationship between FCR and self-esteem (p < 0.001; r = -0.375). In a multivariable logistic regression model, FCR still has a negative correlation with self-esteem (OR, 0.812; 95%CI, 0.734-0.898). Subgroup analysis indicated that the correlation between FCR and self-esteem in cancer survivors was nearly the same in various strata, demonstrating its soundness and stability. This study confirms that elevated self-esteem in individuals surviving cancer may be a protective factor for FCR. Improving the level of self-esteem in cancer survivors can be one of the important directions of clinical interventions for FCR.


[Evaluation of the tears secretion in young patients with cystic fibrosis].
Evaluation of the secretion of the tears in young CF patients. We examined group of 18 CF patients and control group of 18 patients. The patients were evaluated systemically and examined for surface changes and Schirmer test. We observed abnormal Schirmer test in 81% of CF patients. The mean value of Schirmer test was 9.6 +/- 5.5 mm in the CF group and 25.0 +/- 3.0 mm in the control group. The difference was statistically significant. The cystic fibrosis patients have the hyposecretion of tear fluid. Schirmer test may be used in evaluation, diagnosis and treatment of the ocular changes in CF patients.


Efficient Photoinduced Electron Transfer from Pyrene-o-Carborane Heterojunction to Selenoviologen for Enhanced Photocatalytic Hydrogen Evolution and Reduction of Alkynes.
A series of pyrene or pyrene-o-carborane-appendant selenoviologens (Py-SeV2+ , Py-Cb-SeV2+ ) for enhanced photocatalytic hydrogen evolution reaction (HER) and reduction of alkynes is reported. The efficient photoinduced electron transfer (PET) from electron-rich pyrene-o-carborane heterojunction (Py-Cb) with intramolecular charge transfer (ICT) characteristic to electron-deficient selenoviologen (SeV2+ ) (kET = 1.2 × 1010 s-1 ) endows the accelerating the generation of selenoviologen radical cation (SeV+• ) compared with Py-SeV2+ and other derivatives. The electrochromic/electrofluorochromic devices' (ECD and EFCD) measurements and supramolecular assembly/disassembly processes of SeV2+ and cucurbit[8]uril (CB[8]) results show that the PET process can be finely tuned by electrochemical and host-guest chemistry methods. By combination with Pt-NPs catalyst, the Py-Cb-SeV2+ -based system shows high-efficiency visible-light-driven HER and highly selective phenylacetylene reduction due to the efficient PET process.


Treatment with heavy charged particles: systematic review of clinical data and current clinical (comparative) trials.
To analyze relevant data on carbon ion radiotherapy for different tumor indications and to review current clinical trials. All published data on carbon ion radiotherapy were searched for with specific criteria in PUBMED. The terms for search were 'carbon ion and (radiotherapy OR radiation therapy) and (nirs OR chiba OR japan OR itep OR st. petersburg OR PSI OR dubna OR uppsala OR clatterbridge OR loma linda OR nice OR orsay OR itemba OR mpri OR himac OR triumf OR GSI OR HMI OR NCC OR ibmc OR pmrc OR MGH OR infn-lns OR shizuoka OR werc OR zibo OR md anderson OR fpti OR ncc ilsan OR boston OR heidelberg OR tsukuba) NOT in vitro NOT cell culture NOT review[Publication Type] Filters: Humans, English'. The search delivered 273 hits, of which only articles in English including 20 or more patients were included. Case reports were not considered. We subdivided into disease- and site-specific groups. To date, several studies have been performed, however, no randomized trials have been conducted. Therefore, carbon ion radiotherapy must be considered an experimental treatment, and randomized trials comparing modern photon as well as proton treatments are necessary.


Pituitary adenoma and incidental superior hypophyseal aneurysm.
In patients with pituitary adenomas, intra-cranial aneurysms can be an incidental finding, and are usually located outside the pituitary region. The authors describe the multi-modal management of a rare case of an aneurysm of the supraclinoid carotid that encroached into a pituitary macroadenoma.


Impacts of a high-fat diet on the metabolic profile and the phenotype of atrial myocardium in mice.
Obesity, diabetes, and metabolic syndromes are risk factors of atrial fibrillation (AF). We tested the hypothesis that metabolic disorders have a direct impact on the atria favouring the formation of the substrate of AF. Untargeted metabolomic and lipidomic analysis was used to investigate the consequences of a prolonged high-fat diet (HFD) on mouse atria. Atrial properties were characterized by measuring mitochondria respiration in saponin-permeabilized trabeculae, by recording action potential (AP) with glass microelectrodes in trabeculae and ionic currents in myocytes using the perforated configuration of patch clamp technique and by several immuno-histological and biochemical approaches. After 16 weeks of HFD, obesogenic mice showed a vulnerability to AF. The atrial myocardium acquired an adipogenic and inflammatory phenotypes. Metabolomic and lipidomic analysis revealed a profound transformation of atrial energy metabolism with a predominance of long-chain lipid accumulation and beta-oxidation activation in the obese mice. Mitochondria respiration showed an increased use of palmitoyl-CoA as energy substrate. APs were short duration and sensitive to the K-ATP-dependent channel inhibitor, whereas K-ATP current was enhanced in isolated atrial myocytes of obese mouse. HFD transforms energy metabolism, causes fat accumulation, and induces electrical remodelling of the atrial myocardium of mice that become vulnerable to AF.


Effects of emerging contaminants and heavy metals on variation in bacterial communities in estuarine sediments.
Emerging contaminants (ECs) and heavy metals (HMs) are universally present together in estuarine sediments; despite this, their effects on microbial communities have been widely studied separately, rather than in consort. In this study, the combined effects of ECs and HMs on microbial communities were investigated in sediments from 11 major river estuaries around the Bohai Sea, China. Proteobacteria, Bacteroidetes, and Firmicutes were the dominant phyla in the sediments. Using Shannon indices, total phosphorus and total organic carbon were shown to affect microbial community structure. Redundancy analysis of microbial variation implicated Cd and As as the greatest pollutants, followed by Mn, Fe, Zn and Cu; no impacts from galaxolide (HHCB) and tonalide (AHTN) were found. Correlation analysis demonstrated that the concentration of ECs increased the abundance of certain bacteria (e.g., Haliangium, Altererythrobacter, Gaiella and Erythrobacter), and therefore these can be used as potential contamination indicators. Shannon indices and Chao1 indices showed that there were differences in the richness and diversity of bacterial communities in the sediments of 11 rivers. The principal coordinate analysis displayed higher similarity of bacterial community composition in estuarine sediments in Liaoning province than other regions. The results can be used to predict changes in estuary ecosystems to maintain their ecological balance and health.


Proposing a sustainable urban 3D model to minimize the potential risk associated with green infrastructure by applying engineering tools.
The goal of this study is to estimate the potential risk of exposure to urban green infrastructure by calculating and improving of AIROT index, adding meteorological factors as wind direction and updating the index to be more accurate for smaller urban green areas. To achieve this objective, BIM methodology has been applied by creating a 3D BIM model from the reality capture of a street with LiDAR. The BIM model contains the parametric data needed to apply AIROT index and it allows to map results in a graphic environmental sustainability study. The importance of location of green infrastructure is one of main conclusions obtained in order to minimize aerobiological risks in future new buildings or even in maintenance tasks of urban green infrastructure. A valuable result obtained from the developed methodology are walk simulations in the 3D model with the aim to identify high risk of potential exposure of urban green infrastructure with allergenic interest for allergic patients in order to supply health itineraries of pedestrians in a proposal of Smart City.


Comparison of β-endorphin, Lactate and Cortisol Concentrations in Winning and Losing Racehorses.
Understanding the physiological and biochemical changes in racehorses can be invaluable. Accurate information in this area could result in better understanding of needs of sport horses. The aim of this study was to prove the hypothesis that biochemical changes could influence the outcome of competitions. In this study, β-endorphin was evaluated as an indicator of analgesia, lactate as an indicator of fatigue and cortisol as an indicator of stress in the first two horses and the last ones that cross the finish line. This study was performed on 44 horses participating in the 1000-meter national championship. In Group 1, 22 winners and second place horses were included; for Group 2, 22 last and penultimate horses were included. Blood samples were obtained in the doping room after race (T0) and 20 minutes after finishing (T20). Results for beta endorphin at T0 and T20 were higher (P > 0.05) for Group 1 compared to Group 2; on the contrary, lactate concentration was lower (P > 0.05) for Group 1 than Group 2 at T0 and T20. However, differences (P < 0.05) were obtained within groups at T0 and T20 for beta endorphin and lactate concentrations. No significant differences were found for cortisol concentration.The results of this study showed that winning horses had higher levels of β-endorphin and lower levels of lactate than losers. Further and deeper experimental studies are needed to prove the hypothesis that biochemical changes could influence the outcome of competitions.


Bioelectrochemical Purification of Biomass Polymer Derived Furfural Wastewater and Its Electric Energy Recovery.
With the increasing environmental pollution caused by waste polymers, the conversion of polymer components in biomass into valuable products is of great significance for waste management and resource recovery. A two-stage microbial fuel cell (MFC) was used to treat furfural wastewater in this study. The maximum output voltage was 240-250 mV and the power generation time in an operation cycle was 286 h. The degradation efficiency of furfural reached 99-100% (furfural concentration at 300-3000 mg/L) and was slightly reduced to 91% at 7000 mg/L. In addition, the BOD/COD ratio of the furfural wastewater increased from 0.31 to 0.48 after MFC processing. The molecular analysis of the anodic bacterial isolates indicated that the phylogenetic bacterial mixture was dominated by five active anaerobic bacteria with a similarity percentage above 99% for each strain: Burkholderia (B. burdella), Clostridium sensu stricto (Cymbidaceae), Klebsiella (Klebsiella), Ethanoligenens (anaerobic genus), and Acidocella (anaerobic genus); the mixture exhibited good properties to carry out bioelectricity generation in the microbial fuel cell. This indicates that the MFC has effectively degraded furfural for pollutant removal and power generation and is a promising clean method to treat furfural pollution in industry wastewater.


Anti-atherosclerotic efficacy of olmesartan.
The possible inhibition of lipid deposition into vascular tissues by a novel angiotensin II type 1 receptor antagonist, olmesartan, was investigated in a primate high-cholesterol model. Twelve monkeys that were fed a high-cholesterol (4% cholesterol and 6% corn oil) diet for 6 months were divided into two groups: one group was given olmesartan medoxomil (10 mg/kg per day), and the other group was given no medication. A further control group of six monkeys was fed a normal diet throughout the study. The level of low-density lipoprotein (LDL) cholesterol was increased by the high-cholesterol diet, whereas that of high-density lipoprotein (HDL) cholesterol was decreased. Olmesartan decreased the areas of lipid deposition on the aortic surface and intimal cross-section area, but not the mean blood pressure and the levels of LDL and HDL cholesterol. The relaxation response of isolated carotid arteries to acetylcholine was suppressed in the high-cholesterol group, but this was improved by olmesartan. Olmesartan inhibited the accumulation of macrophages in the intimal layer. Serum levels of transforming growth factor (TGF)-beta1, macrophage colony-stimulating factor (M-CSF) and intracellular adhesion molecule (ICAM)-1 were increased in monkeys fed the high-cholesterol diet, but they were suppressed by olmesartan, although the decrease was not significant. Olmesartan reduced lipid deposition, accompanied by the improvement of vascular functions and the inhibition of macrophage accumulation in the intimal layer and showed a trend towards the suppression of serum TGF-beta1, M-CSF and ICAM-1.


Ultrafast labeling and high-fidelity imaging of mitochondria in cancer cells using an aggregation-enhanced emission fluorescent probe.
An aggregation-enhanced emission mitochondrial probe, LIQ-3, was developed for ultrafast labeling within one minute and for distinguishing cancer cells from normal cells. Furthermore, the probe revealed high-fidelity tracking of mitochondria in a three-dimensional localization with advantages that include a specific targeting capacity and a high signal-to-noise ratio.


Isolation and anticoagulant properties of a new sulfated xylan: comparison with heparin and a sodium pentosan polysulfate (SP-54).
Larchwood xylan was purified by diaminoethylaminoethyl (DEAE) cellulose chromatography, sulfated by heating with chlorosulfonic acid -pyridine complex and the sulfated polysaccharide was isolated by epichlorohydrin triethanolamine (ECTEOLA) cellulose chromatography as the sodium salt. It's molecular weight was approximately 25000 and the specific rotation was [alpha]D 20-70 degrees. Electrophoretic analysis of the sulfated xylan along with heparin and SP-54 using lithium acetate-agarose technique showed that the charge density of the xylan sulfate was similar to heparin and it moved as a single component in contrast to commercial heparin which resolved into two while SP-54 moved at the same rate as the marker dye. Anticoagulant properties of the sulfated xylan were compared with heparin and SP-54 by studying its effect on the activated partial thromboplastin time (APTT), prothrombin time (PT) and the thrombin time (TT) using pooled normal human plasma. The sulfated xylan was more effective than SP-54 in delaying coagulation by all the three measurements while it was less active than heparin in inhibiting APTT and PT.


Targeting gasdermin E in neurodegenerative diseases.
Neel et al. identified pathophysiologic clues linking gasdermin-E (GSDME) with frontotemporal dementia and amyotrophic lateral sclerosis.1 Therapeutic studies targeting GSDME may provide a viable approach for neurodegenerative diseases.


Tinidazole in the treatment of trichomoniasis, giardiasis and amoebiasis. Report of a multicentre study.
An open multicentre trial was undertaken in 8 countries to assess the efficacy and tolerance of short course treatment regimens of tinidazole Trichomoniasis: of 859 patients with trichomonal vaginitis given a 2g single dose of tinidazole 717 (95.2%) were cured. Side-effects occurred in 82 patients (9.5%) and in 12 were regarded as severe. Giardiasis: of 74 children with symptomatic giardiasis given tinidazole in a single dose of approximately 50 mg/kg body weight, 65 (88%) were parasitologically and symptomatically cured. 2 (2.7%) complained of side-effects, none of which was severe. Intestinal amoebiasis: of 502 patients, comprising 458 adults and 44 children, with symptomatic intestinal amoebiasis who received tinidazole as a single daily dose on 2 to 3 consecutive days, 477 (95%) were parasitologically cured with complete or marked improvement in symptoms. Side-effects occurred in 50 patients (10%) and in 11 were regarded as severe. Amoebic liver abscess: 82 patients were given tinidazole in single doses of 2g on 3 consecutive days. The response was rated as excellent in 60 and good in 17 (overall cure 93.9%). Side-effects occurred in 9 patients (10.9) and in 2 were regarded as severe. These findings confirm the efficacy and tolerance of short regimens of tinidazole given in single daily doses.


Management of delayed post-pancreaticoduodenectomy arterial bleeding: interventional radiological treatment first.
To investigate the diagnosis and treatment of delayed post-pancreaticoduodenectomy arterial bleeding (DPPAB). Records of 336 patients who underwent pancreaticoduodenectomy (PD) between January 2000 and December 2010 were retrospectively analyzed. Detailed data of patients with DPPAB were assessed by a thorough review of medical records. 14 patients developed DPPAB. The mean time interval between the initial surgery and DPPAB was 33 days (range 7-72). Three patients experienced sentinel bleeding 5-8 days before DPPAB. All DPPAB patients had intra-abdominal septic complications before bleeding. The overall prevalence of success of angiography and transcatheter arterial embolization (TAE) was 85.7% (12/14), including 3 patients who achieved complete hemostasis by TAE after unsuccessful re-laparotomy. The prevalence of mortality of DPPAB was 28.6% (4/14). After hemostasis was achieved, intra-abdominal septic complications were controlled by percutaneous catheter drainage or re-laparotomy with drain replacement. Angiography and TAE are recommended as the first-line diagnostic and treatment choice for DPPAB, respectively. Surgical intervention should be preserved to eliminate the cause of bleeding.


Inhibition of cytomegalovirus-stimulated human cell ornithine decarboxylase by alpha-difluoromethylornithine.
1. The relationship between synthesis of putrescine, human cytomegalovirus DNA synthesis, cell DNA synthesis, and human cytomegalovirus replication has been studied. 2. Stimulation of ornithine decarboxylase activity by shifting low serum-arrested whole human embryo cells to high serum medium is inhibited more than 99% by 2.5 mM DL-alpha-difluoromethylornithine. The addition of DL-alpha-difluoromethylornithine to human cells arrested in low serum and subsequently stimulated by the addition of fresh high serum-containing medium, causes a greater percent inhibition of ornithine decarboxylase activity than when the drug is added to growing human cells. 3. Increased ornithine decarboxylase activity produced by infection of low serum-arrested human cells was inhibited by 5.0 mM of DL-alpha-difluoromethylornithine. However, at a concentration of 5.0 mM, neither DL-alpha-methylornithine nor DL-alpha-difluoromethylornithine affected human cytomegalovirus growth or was toxic to these cells. These data suggest that the increased putrescine synthesis produced by infection is not required for virus replication. 4. The addition of 5.0 mM DL-alpha-difluoromethylornithine had no effect on human cytomegalovirus DNA synthesis or human cytomegalovirus-induced stimulation of cell DNA synthesis. However, 5.0 mM DL-alpha-difluoromethylornithine significantly reduced the stimulation of cell DNA synthesis caused by treatment with mock infecting fluid.


AAV-BDNF mediated attenuation of quinolinic acid-induced neuropathology and motor function impairment.
Maintenance and plasticity of striatal neurons is dependent on brain-derived neurotrophic factor (BDNF), which is depleted in the Huntington's disease striatum due to reduced expression and disrupted corticostriatal transportation. In this study we demonstrate that overexpression of BDNF in the striatum attenuates motor impairment and reduces the extent of striatal damage following quinolinic acid lesioning. Transfer of the BDNF gene to striatal neurons using serotype 1/2 adeno-associated viral vectors enhanced BDNF protein levels in the striatum, but induced weight loss and seizure activity following long-term high-level expression. Lower concentration BDNF expression supported striatal neurons against excitotoxic insult, as demonstrated by enhanced krox-24 immunopositive neuron survival, reduction of striatal atrophy and maintenance of the patch/matrix organization. Additionally, BDNF expression attenuated motor impairment in the forelimb use cylinder test, sensorimotor neglect in the corridor food selection task and reversed apomorphine-induced rotational behaviour. Direct correlations were shown for the first time between BDNF-mediated attenuation of behavioural impairment and the integrity of the globus pallidus, seemingly independent from the severity of striatal lesioning. These results demonstrate that BDNF holds considerable therapeutic potential for alleviating both neuropathological and motor function deficits in the Huntington's disease brain, and the critical role of pallidal neurons in facilitating motor performance.


Assessing the in vitro sensitivity with associated drug resistance polymorphisms in Plasmodium vivax clinical isolates from Delhi, India.
The drug resistance of Plasmodium vivax in clinical cases remains largely unknown till date because of the difficulty in diagnosing the resistant P. vivax strains. The present study was undertaken to determine the prevalence of mutant alleles in drug resistance genes viz P. vivax multi-drug resistance (pvmdr-1), chloroquine resistance transporter (pvcrt-o), dihydrofolate reductase (pvdhfr) and dihydropteroate synthase (pvdhps) along with in vitro chloroquine (CQ) sensitivity in P. vivax clinical isolates. During August-October 2017 a total of 86 samples of the febrile patients were screened and 31 samples were found to be positive for P. vivax in Safdarjung hospital, New Delhi. Sequence genotyping of the drug resistance genes was carried out in these P. vivax samples and in vitro CQ susceptibility for 23 isolates was determined by the schizont maturation assay (SMA). The CQ inhibitory concentrations (IC50) for the clinical isolates was found to be in the range of 25.6-176.7 nM. All the 31 clinical isolates analyzed for pvmdr-1 gene, showed mutant alleles and in only two isolates novel mutations at 861 and 898 codons were observed. Sequence analysis of pvcrt-o, pvdhfr and pvdhps genes revealed wild type genotypes in all the 31 studied isolates. The presence of mutations in pvmdr-1 gene and the increase in the CQ IC50 value indicates the possibility of shift in drug tolerance where CQ with primaquine (PQ) is still the first line of treatment for P. vivax malaria in the country. The regular molecular surveillance in P. vivax would provide useful information for the policy makers of the malaria control programme.


Evaluation of RT-qPCR of mouthwash and buccal swabs for detection of SARS-CoV-2 in children and adults.
The use of nasopharyngeal (NP) swabs as a specimen collection method to diagnose SARS-CoV-2 infection is frequently perceived as uncomfortable by patients and requires trained personnel. In this study, detection rate of SARS-CoV-2 in mouthwash samples and buccal swabs were compared in both children and adults. In patients admitted to hospital with confirmed COVID-19 within the previous 72 hours, NP and buccal swabs as well as mouthwash samples were collected. RT-qPCR was performed on all samples. In total, 170 samples were collected from 155 patients (137 adults and 18 children). Approximately 91.7% of the collected NP swabs were positive in RT-PCR compared to 63.1% of mouthwash samples and 42.4% of buccal swabs. Compared to NP swabs, the sensitivity of using mouthwash was 96.3% and 65.4% for buccal swabs in NP swab samples with a CT value <25. With increasing CT values, sensitivity decreased in both mouthwash and buccal swabs. The virus load was highest during the first week of infection, with a continuous decline observed in all three collection methods over time. Mouthwash presents an alternative collection method for detecting SARS-CoV-2 in the case of unfeasible NP swab sampling. Buccal swabs should not be used due to their low sensitivity.


Increased prevalence of cavum septum pellucidum in schizophrenia.
A previous report that the presence of a cavum septum pellucidum was significantly more common in schizophrenic patients than in control subjects (DeGreef et al., 1992) stimulated the present study of an independent but similar population of first admission schizophrenic patients. A high prevalence of this developmental anomaly (44.8%) was found in the patients (n = 85), although it could also be detected in 29.8% of similarly aged control subjects (n = 47). This finding was not associated with corpus callosum, ventricular, or temporal lobe size or asymmetries.


Premotor potentials.
The topic of this article is probably unfamiliar to most electromyographers. These potentials can be seen at high amplification immediately preceding the median or ulnar compound muscle action potential (CMAP) during standard motor nerve conduction studies. A chronological review of premotor potentials is provided. After longstanding debate, eventual determination of their electrophysiologic origin employed analysis of these potentials using volume conduction principles. The significance of premotor potentials is addressed.


Low-grade fibromyxoid sarcoma in labial mucosa: Report of a rare case in a pediatric patient.
Low-grade fibromyxoid sarcoma (LGFMS) is a malignant neoplasm of soft tissue, which occurs mainly in extremities. In this study, we described a rare case of LGFMS arising in the labial mucosa, in a 7-year-old male patient, who presented a painless multilobulated growth with an evolution time of 12 months. An incisional biopsy was performed and microscopic examination revealed a proliferation of bland regular spindle cells in an alternating myxoid and hyalinized stroma. The tumor cells showed a strong cytoplasmic immunoreactivity for MUC4 and low cell proliferation index measured by Ki-67 antibody. The diagnosis of LGFMS was established and the patient was referred for surgical treatment at a pediatric oncology hospital.


Immunohistochemically detected tumor cells in the sentinel lymph nodes of patients with breast carcinoma: biologic metastasis or procedural artifact?
Sentinel lymph node (SLN) biopsy is a new standard of care for patients with breast carcinoma, and allows enhanced pathologic analysis with serial sections and immunohistochemical (IHC) staining for cytokeratins to be performed on a routine basis. However, the significance of SLN micrometastases detected only by IHC is uncertain. Are these tumor cells truly markers of metastatic potential, or simply evidence of passive displacement by preoperative instrumentation of the tumor site? Here we evaluate whether the pattern of SLN metastasis in breast carcinoma is related to the degree of manipulation at biopsy before surgery, independently of other known predictors. Among 4016 consecutive eligible patients with breast carcinoma registered in a prospective SLN database at Memorial Sloan Kettering Cancer Center, we noted patient/tumor characteristics, pathologic status of the SLN (negative, positive by hematoxylin and eosin [H&E], or positive only on IHC), and method of previous biopsy (none, fine-needle aspiration biopsy [FNAB], core needle biopsy, or surgical biopsy). Multivariate analysis showed that the likelihood of an H&E-positive SLN was significantly associated with lymphovascular invasion, tumor size, tumor type, and tumor location, but not with the method of biopsy. In contrast, the likelihood of finding an SLN positive only on IHC was unassociated with any of the four variables above, but was significantly associated with the method of biopsy. After no previous biopsy, FNAB, core needle biopsy, or surgical biopsy, IHC-positive SLN were present in 1.2%, 3.0%, 3.8%, and 4.6% of patients, respectively (P = 0.002). These data suggest that the frequency of IHC-positive SLN in patients with breast carcinoma 1) is unrelated to conventional predictors of lymph node positivity, 2) is increased after instrumentation of the tumor site, and 3) is increased approximately proportionate to the degree of manipulation. A proportion of IHC-positive SLN were present before biopsy and therefore less likely to be artifactual.


Dynamics of serum metabolites in patients with chronic hepatitis C receiving pegylated interferon plus ribavirin: a metabolomics analysis.
Serum samples from patients with chronic hepatitis C were subjected to metabolomics analysis to clarify the pretreatment characteristics of their metabolites and also changes in specific metabolites resulting from antiviral therapy with pegylated interferon plus ribavirin (PegIFN/RBV). The serum levels of low-molecular-weight metabolites in the twenty patients before and 24weeks after completion of PegIFN/RBV therapy were analyzed using capillary electrophoresis and liquid chromatography-mass spectrometry. Ten patients showed a non-virological response (NVR) and 10 achieved a sustained virological response (SVR) with eradication of viremia. The pretreatment levels of tryptophan were significantly higher in the patients of SVR than in those of NVR (p=0.010). The area under the curve (AUC) value of tryptophan calculated from the receiver operating characteristic (ROC) curve for discriminating SVR from NVR was 0.84 (95% confidential interval, 0.66-1.02, p=0.010). The ROC curve of multiple logistic regression model incorporating the pretreatment levels of tryptophan and γ-glutamate-arginine showed that the AUC value was highly significant (AUC=0.92, 95% confidential interval, 0.79-1.05, p=0.002). Twenty four weeks after completion of treatment, the levels of γ-glutamyl dipeptides, glutamic acid, 5-oxoproline, glucosamine and methionine sulfoxide were decreased, whereas those of 5-methoxy-3-indoleacetate, glutamine, kynurenine and lysine were increased significantly (p<0.05) in both the NVR and SVR patients. The pretreatment serum levels of certain metabolites including tryptophan are associated with the response to PegIFN/RBV therapy. PegIFN/RBV therapy can ameliorate the oxidative stress responsible for glutathione metabolism.


A review of interferon use in patients with relapsing remitting multiple sclerosis in the Canterbury region, New Zealand: 2000-2006.
We report a retrospective medical chart review of 104 patients resident in Canterbury and surrounding districts with relapsing remitting multiple sclerosis (RRMS), who received funded interferon-beta between 2000 and 2006. The aim of the study was to review relapse rates, Expanded disability status scale (EDSS) scores and intravenous methylprednisolone (IVMP) use in the 2-year period before, and following, the initiation of interferon-beta therapy. Demographic analysis showed that the age at entry, duration of disease and EDSS at entry were each greater than in the landmark clinical trials. Relapse rates and usage of IVMP decreased when compared to the 2 years prior to treatment.


C57BL/6J and B6129F1 Embryo Transfer: Unilateral and Bilateral Transfer, Embryo Number and Recipient Female Background Control for the Optimization of Embryo Survival and Litter Size.
Embryo transfer (ET) is a common procedure in rodent facilities. Optimizing this technique may help to reduce the number of animals, but little information is available regarding wild type strains and the conditions that affect embryo transfer. To explore this theme, 2-cell C57BL/6J embryos were transferred after overnight culture of freshly collected zygotes using different conditions: unilateral transfers using a total of 6, 8, 12, 15, 20 and 25 embryos were performed initially; then, this strain was also used for bilateral transfers using a total of 6, 12 and 20 embryos equally divided by the two oviducts. Groups of 25 embryos were not tested for the bilateral technique, since this condition produced the lower success rate when using the unilateral technique and 20 embryos would still represent a large number of embryos. A group of 2-cell B6129F1 embryos was also transferred using unilateral and bilateral ET with 6, 12 and 20 embryos. Crl:CD1(ICR) were used as recipient females for non-reciprocal transfers and C57BL/6J were used to test reciprocal transfers (only tested for six C57BL/6J unilateral transfers). Unilateral transfers using C57BL/6J mice produced higher success rates using six embryos, compared to the other groups transferred unilaterally (p-values between 0.0001 and 0.0267), but the mean number of pups per litter was not different among groups. Bilateral transfer produced higher number of pups when 20 embryos were divided by the two oviducts compared to six (p = 0.0012) or 12 (p = 0.0148) embryos, but with no differences in success rates. No statistical differences were found between the groups of B6129F1, but better results were obtained on bilateral transfers using a total of six embryos. For the strain tested (C57BL/6J), the uterine environment (Crl:CD1(ICR) or C57BL/6J recipient) does not impact the outcome of the technique. These results complement previous work published using genetically engineered mice strains and show that unilateral transfers using low number of embryos (6), produce better outcomes when compared to bilateral or unilateral transfers using more embryos. It also highlights differences between the outcome of bilateral transfers in the two strains tested. A set of historical data of genetically engineered mice at a C57BL/6J background was also included, confirming that lower embryo numbers are related to higher success rates. Together, the outcome of these experiments can be important to reduce the number of recipient and donor females, optimize embryo transfers and improve animal welfare discouraging the use of a more invasive technique.


Development and validation of the sleep assessment instrument for older adults with pain.
The co-occurrence of chronic pain and sleep disturbance contribute to a significant functional and social impact in older adults. However, there are no validated instruments to measure sleep disturbance and pain in this population that could be used to screen or diagnose individuals or monitor treatment effectiveness. Our aim was to develop and validate a brief, practical, and comprehensive tool to assess the impact of co-occurring pain and sleep disturbance in older adults. Development and validation of a measurement tool for assessing pain and sleep in older adults consisting of seven items. We applied the "Sleep Assessment Instrument for Pain in older adults" (SAIOAP) in a sample of 100 older individuals. A Cronbach's alpha of 0.602 indicated a moderate level of reliability, and item-total correlations of ≥0.4 for all items indicated good homogeneity. There were statistically significant correlations between the SAIOAP and sleep quality (PSQI, r=61.5), pain intensity (VNS, r=30.5), the multidimensional impacts of pain (GPM, r=40.5), depression (GEAP, r=45.5), comorbidity (r=27.9), and medication use (r=30.4). A ROC curve indicated a sensitivity of 73.2% and a specificity of 79.1% in relation to the prediction of sleep disturbances associated with pain in older adults. The SAIOAP presented adequate metric properties and was demonstrated to be a simple and practical tool for the assessment of the impact of pain on sleep in older adults.


The role of Müller cell glucocorticoid signaling in diabetic retinopathy.
Diabetic retinopathy (DR) is a sight-threatening complication associated with the highly prevalent diabetes disorder. Both the microvascular damage and neurodegeneration detected in the retina caused by chronic hyperglycemia have brought special attention to Müller cells, the major macroglia of the retina that are responsible for retinal homeostasis. Given the role of glucocorticoid signaling in anti-inflammatory responses and the almost exclusive expression of glucocorticoid receptors (GRs) in retinal Müller cells, administration of corticosteroid agonists as a potential treatment option has been widely studied. Although these approaches have been moderately efficacious in treating or de-escalating DR pathomechanisms, there are various side effects and gaps of knowledge with regard to introducing exogenous glucocorticoids to the diseased retina. In this paper, we provide a review of the literature concerning the available evidence for the role of Müller cell glucocorticoid signaling in DR and we discuss previously investigated approaches in modulating this system as possible treatment options. Furthermore, we propose a novel alternative to the available choices of treatment by using gene therapy as a tool to regulate the expression of GR in retinal Müller cells. Upregulating GR expression allows for induced glucocorticoid signaling with more enduring effects compared to injection of agonists. Hence, repetitive injections would no longer be required. Lastly, side effects of glucocorticoid therapy such as glucocorticoid resistance of GR following chronic exposure to excess ligands or agonists can be avoided.


Assessment of reliability of CAD-CAM tooth-colored implant custom abutments.
Information is lacking about the fatigue resistance of computer-aided design and computer-aided manufacturing (CAD-CAM) tooth-colored implant custom abutment materials. The purpose of this in vitro study was to investigate the reliability of different types of CAD-CAM tooth-colored implant custom abutments. Zirconia (Lava Plus), lithium disilicate (IPS e.max CAD), and resin-based composite (Lava Ultimate) abutments were fabricated using CAD-CAM technology and bonded to machined titanium-6 aluminum-4 vanadium (Ti-6Al-4V) alloy inserts for conical connection implants (NobelReplace Conical Connection RP 4.3×10 mm; Nobel Biocare). Three groups (n=19) were assessed: group ZR, CAD-CAM zirconia/Ti-6Al-4V bonded abutments; group RC, CAD-CAM resin-based composite/Ti-6Al-4V bonded abutments; and group LD, CAD-CAM lithium disilicate/Ti-6Al-4V bonded abutments. Fifty-seven implant abutments were secured to implants and embedded in autopolymerizing acrylic resin according to ISO standard 14801. Static failure load (n=5) and fatigue failure load (n=14) were tested. Weibull cumulative damage analysis was used to calculate step-stress reliability at 150-N and 200-N loads with 2-sided 90% confidence limits. Representative fractured specimens were examined using stereomicroscopy and scanning electron microscopy to observe fracture patterns. Weibull plots revealed β values of 2.59 for group ZR, 0.30 for group RC, and 0.58 for group LD, indicating a wear-out or cumulative fatigue pattern for group ZR and load as the failure accelerating factor for groups RC and LD. Fractographic observation disclosed that failures initiated in the interproximal area where the lingual tensile stresses meet the compressive facial stresses for the early failure specimens. Plastic deformation of titanium inserts with fracture was observed for zirconia abutments in fatigue resistance testing. Significantly higher reliability was found in group ZR, and no significant differences in reliability were determined between groups RC and LD. Differences were found in the failure characteristics of group ZR between static and fatigue loading.


Bursting firing of action potentials in central snail neurons elicited by d-amphetamine: role of the electrogenic sodium pump.
The effects of d-amphetamine on central neurons were studied electrophysiologically in the identifiable RP4 neuron of the African snail, Achatina fulica Ferussac. d-Amphetamine elicited bursting activity from the central RP4 neuron in a concentration-dependent manner. The bursting activity was not blocked in a high magnesium (30 mM) medium, or after a continuous perfusion of propranolol, prazosin, haloperidol, phenobarbital, hexamethonium, d-tubocurarine, atropine, or calcium-free solution containing EDTA or verapamil. These results suggested that the bursting activity elicited by d-amphetamine was not due to: (1) the synaptic effects of neurotransmitters; or (2) the cholinergic or adrenergic receptors of the excitable membrane. However, the bursting activity elicited by d-amphetamine was blocked in the presence of ouabain or in the medium containing potassium-free, low sodium solutions. d-Amphetamine did not elicit the bursting activity of the LP4 neuron in the same ganglia preparation, and did not alter the GABA-elicited currents of the snail neuron. It is concluded, therefore, that d-amphetamine induced a potassium- and sodium-dependent bursting activity of central neurons. The bursting activity of the central neuron may be associated with the sodium pump of the neuron.


Feasibility of a multidisciplinary group videoconferencing approach for chronic low back pain: a randomized, open-label, controlled, pilot clinical trial (EN-FORMA).
Low back pain is a common condition that becomes even more prevalent with aging. A non-pharmacological multidisciplinary approach for chronic non-specific low back pain (CNSLBP) has been recommended, but integrating different healthcare professionals is challenging. A multidisciplinary group videoconferencing approach (MGVA) can be helpful. Our aim was to provide evidence on MGVA's feasibility in managing CNSLBP and its impact on clinical practice. We conducted an open-label, randomized, controlled, parallel-group pilot clinical trial with CNSLBP patients irresponsive to conservative treatment. Patients between 18 and 67 years of age were randomly assigned (1:1) to either Standard-of-Care + MGVA (experimental group) or Standard-of-Care alone (control group). MGVA consisted of integrated sessions for physical rehabilitation/physiotherapy, psychology, and social work treatments. The control group received standard clinical practice treatment. The feasibility was assessed by the number of study procedures completed to at least 80% as planned. The impact on clinical practice was evaluated by the number of patients who changed their status from "candidate" to "non-candidate" to low back surgery as the treatment of choice for CNSLBP. The SF-36, Oswestry Disability Index, and TMMS-24 questionnaires were used. We performed a whole population-based descriptive analysis. We included 20 patients, but only 18 were randomized (2 withdrew consent before randomization). The mean (SD) age was 53.1 (5.9) years, and mostly women (13/18); 7 were actively employed. In the experimental group, 6/9 (67%, 95%CI:35-88%) patients attended at least 80% of the scheduled procedures, while in the control group, 8/9 (89%, 95%CI:57-98%) did. Additionally, 1 out of 4 (25%) patients in the experimental group changed their status from "candidate" for low back surgery to "non-candidate". None of the 2 patients in the control group did so. We found differences between groups in the SF-36 mental health component (p-value:0.061), Oswestry Disability Index (p-value:0.032), and TMMS-24 Repair component (p-value:0.014) at the end of the trial favoring MGVA. The multidisciplinary group videoconferencing approach to managing chronic non-specific low back pain was feasible, suggesting overall beneficial effects on patients' health and could play a role in changing a patient's status from "candidate" to "non-candidate" for surgery. NCT05093543 (ClinicalTrials.gov), first registered in 26/10/2021.


Effect of branched chain amino acid enrichment of total parenteral nutrition on nitrogen sparing and clinical outcome of sepsis and trauma: a prospective randomized double blind trial.
Administration of extra branched chain amino acids (BCAA) has been associated with a nitrogen sparing effect in septic and traumatized patients. Whether nitrogen sparing is associated with decreased morbidity and mortality rates is unknown. We therefore undertook a prospective, randomized, double blind trial investigating the effects of BCAA enrichment of a total parenteral nutrition (TPN) regimen on nitrogen balance, 3-methylhistidine excretion, morbidity as evidenced by disturbances in organ function, severity of sepsis and mortality. One hundred and one patients entered the study; 52 received a standard TPN solution and 49 a BCAA-enriched solution. Both groups received 30 kcal kg-1 body-weight, 15 per cent fat calories and 0.17 g nitrogen kg-1 body-weight. In the BCAA-enriched group, patients received 0.56 g BCAA kg-1 body-weight (50.2 per cent BCAA). Standard group patients received 0.18 g BCAA kg-1 body-weight (15.6 per cent BCAA). Nitrogen balances and 3-methylhistidine excretion were not significantly different between groups. Although morbidity scores tended to decrease during the study no difference was observed between groups. Mortality (early or late), sepsis or stress-related, did not differ significantly between groups. We were not able to confirm the reported beneficial effects of BCAA-enriched TPN solutions for use in septic and traumatized patients.


Safety and efficacy of tenofovir alafenamide in liver transplant recipients: A single center experience.
Tenofovir disoproxil fumarate (TDF) is frequently used for treatment of and prophylaxis against reactivation of hepatitis B virus (HBV) after liver transplant (LT). Because TDF can lead to renal impairment and a decrease in bone mineral density (BMD), the prodrug tenofovir alafenamide (TAF) may be considered a viable alternative with fewer adverse effects. Only limited information is available about the use of TAF for LT recipients. We report a European single-center experience with TAF as treatment for LT patients. This retrospective analysis involved 29 LT recipients receiving standard immunosuppressants (mainly calcineurin inhibitors). Demographic and clinical data were documented at baseline upon switch to TAF and at various time points thereafter. None of the patients experienced HBV reactivation after the switch to TAF. Liver and renal function remained stable. Drug levels of immunosuppressive agents did not change significantly after the switch. After 1 year, 22 patients were still taking TAF; two patients had been lost to follow-up; one patient had died; and four patients had discontinued therapy because of TAF-related adverse effects. No serious adverse effects were reported. Tenofovir alafenamide exhibits high antiviral efficacy and a good safety profile for LT recipients. Still, the safety and tolerability of TAF for organ transplant patients should be evaluated in larger cohorts.


Vertebral artery dissection complicated by basilar artery occlusion.
Acute basilar artery occlusion (ABAO) is an infrequent but potentially fatal complication that can cause strokes in both adults and children. Traumatic vertebral artery dissection (VAD) is one of the most common causes of ABAO in young patients. We present a case of an 11-year-old boy with VAD complicated by basilar artery occlusion 2 days after a fight with classmates that caused severe neurological deficits. He did not have any direct head trauma or concomitant risk factors. Clinical symptoms included nausea, vomiting, and rapid alteration of consciousness. Magnetic resonance imaging showed total occlusion of the basilar artery, and angiography confirmed VAD from the third to the fourth segments. A history of such subtle precipitating events should be noted when diagnosing young patients with brainstem strokes. A delay in the diagnosis of ABAO is frequently due to misleading symptoms and signs and the lack of awareness of this rare condition.


Successive sheep grazing reduces population density of Brandt's voles in steppe grassland by altering food resources: a large manipulative experiment.
Livestock grazing has shaped grassland ecosystems around the world. Previous studies indicated grazing showed various impacts on small rodents; however, most studies were conducted over 1-2 years without controlling for confounding factors such as immigration/emigration and predation in rodents. Brandt's voles (Lasiopodomys brandtii) are generally recognized as pests because of food overlap with domestic herbivores, but are also important for biodiversity conservation because they provide nests or food to many birds. Fully understanding the ecological relationship between domestic herbivores and small mammals is essential to making ecosystem management decisions. To address these needs, we carried out a field experiment during the period 2010-2013 to assess the effects of sheep grazing on vegetation and the population density of Brandt's voles along a gradient of three grazing intensities by using 12 large-scale enclosures. Responses of Brandt's voles to livestock grazing varied with grazing intensity and year. As compared to the control group, sheep grazing had no effect on vole abundance in the first year but an overall negative effect on vole abundance in the following 3 years. Successive grazing caused decreases in survival and male body mass of voles, but had no significant effect on fecundity. Negative effects of grazing were associated with a grazing-induced deterioration in both food quantity (reflected by biomass and cover of less-preferred plants), and food quality (measured by tannin and total phenol content). Our findings highlight the urgent need for more flexible management of yearly rotational grazing to optimize livestock production while maintaining species diversity and ecosystem health.


Complications of rhBMP-2 utilization for posterolateral lumbar fusions requiring reoperation: a single practice, retrospective case series report.
Recombinant human bone morphogenetic protein-2 (rhBMP-2) (INFUSE, Medtronic, Memphis, TN, USA) has been used off-label for posterolateral lumbar fusions for many years. The goal of this study was to evaluate the complications requiring reoperation associated with rhBMP-2 application for posterolateral lumbar fusions. During a 7-year period of time (2002-2009), all patients undergoing lumbar posterolateral fusion using rhBMP-2 (INFUSE) were retrospectively evaluated within a large orthopedic surgery private practice. A total of 1,158 consecutive patients were evaluated with 468 (40.4%) males and 690 (59.6%) females. Complications related to rhBMP were defined as reoperation secondary to symptomatic failed fusion (nonunion), symptomatic seroma formation, symptomatic reformation of foraminal bone, and infection. Inclusion criteria were posterolateral fusion with rhBMP-2 implant and age equal to or older than 18 years. Surgical indications and treatment were performed in accordance with the surgeon's best knowledge, discretion, and experience. Patients consented to lumbar decompression and arthrodesis using rhBMP-2. All patients were educated and informed of the off-label utilization of rhBMP-2. Patient follow-up was performed at regular intervals of 2 weeks, 6 weeks, 12 weeks, 6 months, 1 year, and later if required or indicated. Average age was 59.2 years, and body mass index was 30.7 kg/m². Numbers of levels fused were 1 (414, 35.8%), 2 (469, 40.5%), 3 (162, 14.0%), 4 (70, 6.0%), 5 (19, 1.6%), 6 (11, 0.9%), 7 (7, 0.6%), 8 (4, 0.3%), and 9 (2, 0.2%). Patients having complications requiring reoperation were 117 of 1,158 (10.1%): symptomatic nonunion requiring redo fusion and instrumentation 41 (3.5%), seroma with acute neural compression 32 (2.8%), excess bone formation with delayed neural compression 4 (0.3%), and infection requiring debridement 26 (2.2%). Nonunion was related to male sex and previous BMP exposure. Seroma formation was significantly higher in patients with higher doses of rhBMP-2 (p=.050) and with more than 12 mg of rhBMP-2 (χ(2)=0.025). Bone reformation and neural compression at the laminectomy and foraminotomy sites occurred in a delayed fashion. Infection was associated with obesity and respiratory disease. Infections were noted with a greater BMP dose (p<.001), more than 12 mg (χ(2)<0.001), fusion more than three levels (χ(2)<0.001), and reexposed to BMP (χ(2)=0.023). rhBMP-2 utilization for posterolateral lumbar fusions has a low symptomatic nonunion rate. Prior rhBMP-2 exposure and male sex were related to symptomatic nonunion formation. rhBMP-2-associated neural compression acutely with seroma formation and delayed with foraminal bone formation is concerning and associated with higher rhBMP-2 concentrations.


An unusual filling defect in the ureter.
A case of benign ureteric haemangioma is reported. A conservative approach is recommended when the kidney is capable of function.


Fluorescein as a model molecular calculator with reset capability.
The evolution of molecules capable of performing boolean operations has gone a long way since the inception of the first molecular AND logic gate, followed by other logic functions, such as XOR and INHIBIT, and has reached the stage where these tiny processors execute arithmetic calculations. Molecular logic gates that process a variety of chemical inputs can now be loaded with arrays of logic functions, enabling even a single molecular species to execute distinct algebraic operations: addition and subtraction. However, unlike electronic or optical signals, the accumulation of chemical inputs prevents chemical arithmetic systems from resetting. Consequently, a set of solutions is required to complete even the simplest arithmetic cycle. It has been suggested that these limitations can be overcome by washing off the input signals from solid supports. An alternative approach, which does not require solvent exchange or incorporation of bulk surfaces, is to reset the arithmetic system chemically. Ultimately, this is how some biological systems regenerate. Here we report a highly efficient and exceptionally simple molecular arithmetic system based on a plain fluorescein dye, capable of performing a full scale of elementary addition and subtraction algebraic operations. This system can be reset following each separate arithmetic step. The ability to selectively eradicate chemical inputs brings us closer to the realization of chemical computation.


[Definition and use of the terms "ultimate factor", "proximate factor" and "zeitgeber"].
The use of the terms, which have not always been employed uniformly throughout the literature, is discussed and an exact definition is tried. Application of the terms "ultimate" and "proximate factors" should not be restricted to annual and circadian periodicities. Instead they should be used for all temporally or spatially restricted processes for which a difference between selecting and regulating mechanisms is to be found. "Ultimate factors" are environmental factors which in the course of evolution have led, through natural selection, to the relevant restriction. Proximate factors may be defined as those external stimuli which initiate or maintain biological processes under most favourable ecological conditions. The term "Zeitgeber" finally, should only be used where endogenous circadian or circennial periodicities are synchronized with environmental changes through external factors. In this sense "Zeitgeber" is merely a special case of "proximate factor".


Polyethylene glycol embedded reduced graphene oxide supramolecular assemblies for enhanced room-temperature gas sensors.
Herein, we present the gas-dependent electrical properties of a reduced graphene oxide nanocomposite. The reduced graphene oxide (rGO) was synthesized by reducing GO with sodium borohydride (NaBH4). As-synthesized rGO was dispersed in DI water containing 1, 2, 3, 4, and 5 wt% polyethylene glycol (PEG) to prepare PEG-rGO supramolecular assemblies. The successful preparation of supramolecular assemblies was verified by their characterization using XRD, FESEM, EDS, TEM, FTIR, and Raman spectroscopy. At room temperature, the gas-dependent electrical properties of these supramolecular assemblies were investigated. The results showed that sensors composed of PEG-rGO supramolecular assemblies performed better against benzene and methanol at 3% and 4% PEG, respectively. However, high selectivity and a wide range of activation energies (∼1.64-1.91 eV) were observed for H2 gas for 4% PEG-modified supramolecular assemblies. The PEG-rGO supramolecular assemblies may be an excellent candidate for constructing ultrahigh-performance gas sensors for a variety of applications due to their high sensitivity and selectivity.


[Multiple primary malignancies of the genitourinary tract--renal cell carcinoma and seminoma testis].
Multiple primary malignancies are tumors which present a definite picture of malignancy, each must be distinct and the possibility that one is a metastasis of another must be excluded. Authors present rare case of renal cancer associated with testicular cancer.


Growth kinetics of an Indian isolate of highly pathogenic porcine reproductive and respiratory syndrome virus in MARC-145 cells.
The aim of the present study was to understand the replication kinetics of an Indian isolate of highly pathogenic porcine reproductive and respiratory syndrome (PRRS) virus (Ind-297221) in MARC-145 cells infected at different multiplicity of infection (MOI) of 1.0, 0.1, 0.01 and 0.001. PRRSV titre in the infected cell fraction and the culture supernatant harvested at different intervals (12, 36, 48, 72, 96 and 120 h) post infection (hpi) was estimated by immunoperoxidase monolayer assay. Viral RNA copy numbers were quantified by TaqMan RT-PCR. PRRS virus could be detected first in intracellular fraction at 12 hpi in cells infected at 1.0 MOI, whereas in the extracellular fraction, earliest detection was at 36 hpi. Highest PRRSV titre of 1.3 × 105.0 TCID50/mL was achieved in 0.01 and 0.001 MOI groups at 96 hpi. Infection with 0.01 MOI resulted in the maintenance of maximum titre up to 120 hpi. The maximum viral copy numbers observed was 3.15 × 107.0 in 0.1 MOI group at 120 hpi in culture medium. The results of the study showed that MARC-145 cells infected with Indian PRRSV at 0.01 MOI and harvested in 96-120 hpi was found to be optimum for obtaining maximum virus yield and hence can be used for bulk propagation of the virus.


Effect of estradiol benzoate on the pituitary-gonadal axis in the intact male rat.
Adult male rats were treated with 50 microgram of estradiol benzoate daily for 28 days. The sex organs and pituitary weights showed changes classically associated with estrogen treatment. Both plasma and testicular testosterone levels diminished to almost trace level 24 hours after the first estrogen injection and remained at that level throughout the experimental period. The pituitary LH levels and concentration showed a progressive decline; however, the plasma LH levels remained unchanged throught the experimental period. These findings suggest that in intact adult males, estrogen affects testicular testosterone production directly. These observation suggests the necessity for reevaluation of the current concepts concerned with testis-pituitary feedback mechanisms.


Prevalence of hereditary hemochromatosis in 16031 primary care patients.
Despite evidence from screening studies in northern European populations, the prevalence of hemochromatosis in primary care populations in the United States remains speculative. To establish the feasibility of screening for hemochromatosis and to estimate the prevalence of hemochromatosis in a large primary care population. Cross-sectional prevalence study. 22 primary care practices in the Rochester, New York, area. 16031 ambulatory patients without a previous diagnosis of hemochromatosis. Serum transferrin saturation screening tests were offered to all adult patients in participating primary care practices. Patients with a serum transferrin saturation of 45% or more on initial testing had a serum transferrin saturation test done under fasting conditions and had serum ferritin levels measured. Those who had a fasting serum transferrin saturation of 55% or more and a serum ferritin level of 200 microg/L or more with no other apparent cause were presumed to have hemochromatosis and were offered liver biopsy to confirm the diagnosis. 25 patients had biopsy-proven hemochromatosis; 22 patients met the clinical criteria for hemochromatosis but declined liver biopsy and were classified as having clinically proven hemochromatosis; and 23 patients had a serum transferrin saturation of 55% or more with no identifiable cause, indicating probable hemochromatosis. The prevalence of clinically proven and biopsy-proven hemochromatosis combined was 4.5 per 1000 (95% CI, 3.3 to 5.8 per 1000) in the total sample and 5.4 per 1000 (CI, 4.0 to 7.1 per 1000) in white persons. The prevalence was higher in men than in women (ratio, 1.8:1). Hemochromatosis is relatively common among white persons. Routine screening of white persons for hemochromatosis should be considered by primary care physicians.


Removal of dichloromethane from ground and wastewater: a review.
Dichloromethane (DCM) is a toxic volatile compound which is found in the ground waters and wastewaters of the pharmaceutical, chemical, textile, metal-working and petroleum industries. DCM inhibits the growth of aquatic organisms, induces cancer in animals and is potentially carcinogenic for humans. This article aims to review existing water treatments for DCM removal, focusing on recent technological advances. Air stripping, adsorption and pervaporation were found to be effective in separating DCM from water with a process efficiency of about 99%, 90% and 80% respectively. Electrocatalysis over Cu-impregnated carbon fiber electrode, photo irradiation over TiO₂ and photo-Fenton process led to the complete decomposition of DCM. Aerobic and anaerobic water treatment achieved 99% and 95% removal of DCM respectively. The maximum efficiencies observed for acoustic cavitation, radiolysis and catalytic degradation of CH₂Cl₂ were 90%, 92% and 99% respectively. Ozonation and persulfate oxidation showed lower DCM degradation efficiencies, not exceeding 20%. Further combination of different water treatment methods will further increase DCM degradation efficiency.


Endothelial NOX4 aggravates eNOS uncoupling by decreasing dihydrofolate reductase after subarachnoid hemorrhage.
Endothelial malfunction is a major contributor to early or delayed vasospasm after subarachnoid hemorrhage (SAH). As a representative form of endothelial dysfunction, endothelial nitric oxide synthase (eNOS) uncoupling leads to a reduction in nitric oxide (NO) generated by endothelial cells. In this study, we investigated how the interaction between endothelial NOX4 (nicotinamide adenine dinucleotide phosphate oxidase 4) and DHFR (dihydrofolate reductase) contributes to eNOS uncoupling after SAH. Setanaxib and the adeno-associated virus (AAV) targeting brain vascular endothelia were injected through the tail vein and the expression and localization of proteins were examined by western blot and immunofluorescence staining. The NO content was measured using the NO assay kit, and laser speckle contrast imaging was used to assess cortical perfusion. ROS (reactive oxygen species) level was detected by DHE (dihydroethidium) staining, DCFH-DA (2',7'-dichlorofluorescin diacetate) staining and H2O2 (hydrogen peroxide) measurement. The Garcia score was employed to examine neurological function. Setanaxib is widely used for its preferential inhibition for NOX1/4 over other NOX isoforms. After endothelial NOX4 was inhibited by Setanaxib in a mouse model of SAH, the endothelial DHFR level was significantly elevated, which attenuated eNOS uncoupling, increased cortical perfusion, and improved the neurological function. The protective role of inhibiting endothelial NOX4, however, disappeared after knocking down endothelial DHFR. Our results suggest that endothelial DHFR decreased significantly because of the elevated level of endothelial NOX4, which aggravated eNOS uncoupling after SAH, leading to decreased cortical perfusion and worse neurological outcome.


[Problems of exposure to organic mercury compounds in pregnancy].
On account of oral intake of mercury by contaminated foodstuff (eggs eaten by pregnant women) due to non-observance of legal decisions increased mercury concentrations in the maternal and infantile blood, in the urine and hair of the mother, in the placenta, the amnion, the cord blood and the breast milk could be proved in comparison to non-exposed pregnant women. Clinical and laboratory-chemical parameters did not show any symptoms of an intoxication in mothers and children. Demands for observance of the legal decisions and avoidance of professional dealing are particularly important for pregnant women.


A re-analysis of the iron content of plant-based foods in the United Kingdom.
In the UK contemporary estimates of dietary Fe intakes rely upon food Fe content data from the 1980s or before. Moreover, there has been speculation that the natural Fe content of foods has fallen over time, predominantly due to changes in agricultural practices. Therefore, we re-analysed common plant-based foods of the UK diet for their Fe content (the '2000s analyses') and compared the values with the most recent published values (the '1980s analyses') and the much older published values (the '1930s analyses'), the latter two being from different editions of the McCance and Widdowson food tables. Overall, there was remarkable consistency between analytical data for foods spanning the 70 years. There was a marginal, but significant, apparent decrease in natural food Fe content from the 1930s to 1980s/2000s. Whether this represents a true difference or is analytical error between the eras is unclear and how it could translate into differences in intake requires clarification. However, fortificant Fe levels (and fortificant Fe intake based upon linked national data) did appear to have increased between the 1980s and 2000s, and deserve further attention in light of recent potential concerns over the long-term safety and effectiveness of fortificant Fe. In conclusion, the overall Fe content of plant-based foods is largely consistent between the 1930s and 2000s, with a fall in natural dietary Fe content negated or even surpassed by a rise in fortificant Fe but for which the long-term effects are uncertain.


Low-dose dual-source CT angiography with iterative reconstruction for coronary artery stent evaluation.
The purpose of this study was to evaluate the image quality and diagnostic accuracy of very low-dose, dual-source computed tomography (DSCT) angiography for the evaluation of coronary stents. Iterative reconstruction (IR) leads to substantial reduction of image noise and hence permits the use of very low-dose data acquisition protocols in coronary computed tomography angiography. Fifty symptomatic patients with 87 coronary stents (diameter 3.0 ± 0.4 mm) underwent coronary DSCT angiography (heart rate, 60 ± 6 beats/min; prospectively electrocardiography-triggered axial acquisition; 80 kV, 165 mA, 2 × 128 × 0.6-mm collimation; 60 ml of contrast at 6 ml/s) before invasive coronary angiography. DSCT images were reconstructed using both standard filtered back projection and a raw data-based IR algorithm (SAFIRE, Siemens Healthcare, Forchheim, Germany). Subjective image quality (4-point scale from 0 [nondiagnostic] to 3 [excellent image quality]), image noise, contrast-to-noise ratio as well as the presence of in-stent stenosis >50% were independently determined by 2 observers. The median dose-length product was 23.0 (22.0; 23.0) mGy · cm (median estimated effective dose of 0.32 [0.31; 0.32] mSv). IR led to significantly improved image quality compared with filtered back projection (image quality score, 1.8 ± 0.6 vs. 1.5 ± 0.5, p < 0.05; image noise, 70 Hounsfield units [62; 80 Hounsfield units] vs. 96 Hounsfield units [82; 113 Hounsfield units], p < 0.001; contrast-to-noise ratio, 11.0 [9.6; 12.4] vs. 8.0 [6.2; 9.3], p < 0.001). To detect significant coronary stenosis in filtered back projection reconstructions, the sensitivity, specificity, positive predictive value, and negative predictive value were 97% (32 of 33), 53% (9 of 17), 80% (32 of 40), and 90% (9 of 10) per patient, respectively; 89% (43 of 48), 79% (120 of 152), 57% (42 of 74), and 96% (121 of 126) per vessel, respectively; and 85% (12 of 14), 69% (51 of 73), 32% (11 of 34), and 96% (51 of 53) per stent, respectively. In reconstructions obtained by IR, the corresponding values were 100% (33 of 33), 65% (11 of 17), 85% (33 of 39), and 100% (11 of 11) per patient, respectively; 96% (46 of 48), 84% (129 of 152), 66% (47 of 71), and 98% (127 of 129) per vessel, respectively; and 100% (14 of 14), 75% (55 of 73), 44% (14 of 32), and 100% (55 of 55) per stent, respectively. These differences were not significant. In selected patients, prospectively electrocardiography-triggered image acquisition with 80-kV tube voltage and low current in combination with IR permits the evaluation of patients with implanted coronary artery stents with reasonable diagnostic accuracy at very low radiation exposure.