Redirected T-cell cytotoxicity to epithelial cell adhesion molecule-overexpressing adenocarcinomas by a novel recombinant antibody, E3Bi, in vitro and in an animal model.
To redirect cytotoxic T cells to target a broad range of adenocarcinomas, the authors constructed a novel, recombinant, bispecific antibody, E3Bi, directed at the tumor-associated antigen, epithelial cell adhesion molecule (EpCAM), and the CD3 receptor on T cells. T cells were prepared from healthy blood donors. The cytotoxicity of activated T cells (ATC) redirected to tumor cells by E3Bi was measured with in vitro (51)Cr release assays. In vivo studies were performed in a severe combined immunodeficient (SCID)/Beige mouse xenograft model. Tumor-bearing mice were treated with low doses (1 mg/kg) or high doses (10 mg/kg) of E3Bi along with ATC (2 x 10(9) cells/kg), and treatment efficacy was evaluated both by ex vivo tumor cell survival assay after in vivo treatments and by in vivo tumor growth delay studies. In vitro, targeting the EpCAM-overexpressing human tumor cell lines with E3Bi increased specific cytotoxicity of ATC by > 70% at an effector-to-target ratio of 2.5 (P < 0.001); this cytotoxicity was abolished competitively in the presence of an anti-EpCAM monoclonal antibody. In contrast, E3Bi did not enhance ATC cytotoxicity toward the low EpCAM-expressing tumor cell line. In ex vivo tumor cytotoxicity assays, a significant reduction in tumor cell survival (40% with low-dose E3Bi; 90% with high-dose E3Bi) was observed in E3Bi/ATC-treated mice compared with control mice that were treated with ATC only. In addition, SCID/Beige mice xenografted with LS174T tumors demonstrated a significant tumor growth delay (P = 0.0139) after receiving E3Bi/ATC/interleukin 2 (IL-2) compared with mice that received ATC/IL-2 alone. E3Bi specifically and very efficiently redirected T cells to destroy EpCAM-overexpressing tumors both in vitro and in an animal model. These results suggest a therapeutic utility for E3Bi in the treatment of adenocarcinomas.


Effect of gravity and capillarity on human saliva penetration in coronally unsealed obturated root canals.
The purpose of this study was to evaluate the effect of gravity and capillarity on penetration of human salivary bacteria into the entire length of obturated root canals, and to demonstrate the dye penetration configuration. Fifty single-rooted premolars were decoronated, prepared to a standardized length of 15 mm, instrumented, and randomly divided into two groups (A and B) of 25 teeth each. Each group consisted of experimental (15 samples) and negative and positive controls (five samples each). The experimental groups were obturated with gutta-percha and root canal sealer. The positive control groups were obturated with a single cone of gutta-percha and root canal sealer. The outer surfaces (except for the apical 2 mm) were covered with two layers of nail varnish. An apparatus containing Brain Heart Infusion broth was designed, in which the teeth were placed. The samples in Group A were placed upside down, while Group B was placed normally. The coronal portions of the samples were placed in contact with fresh saliva. The number of days required for bacteria to penetrate the entire length of canals was determined. The samples were then immersed in India ink to determine the dye penetration configuration. Data were analyzed using Student's t-test. The extent of dye penetration was significantly greater in Group B compared to Group A, and they were in a pattern rather than linear form. Gravity and capillarity insignificantly affected bacterial leakage. Although gravity and capillarity did not affect bacterial penetration when applied to the coronal access of endodontically treated teeth, it seems that they can promote penetration of India ink into the canal after the bacterial test on the same tooth.


An improved approach to hydrophilic interaction chromatography of peptides: salt gradients in the presence of high isocratic acetonitrile concentrations.
Hydrophilic interaction chromatography (HILIC) for separations of peptides has been employed infrequently, particularly considering that this technique was introduced over 20 years ago. The present manuscript describes a radical departure from the traditional HILIC elution approach, where separations are achieved via increasing salt (sodium perchlorate) gradients in the presence of high isocratic concentrations (>80%) of acetonitrile, denoted HILIC/SALT. This initial study compared to reversed-phase chromatography (RPC), HILIC and HILIC/SALT for the separation of mixtures of synthetic peptide standards varying in structure (amphipathic α-helix, random coil), length (10-26 residues), number of positively charged residues (+1 to +11) and hydrophilicity/hydrophobicity. Results showed a marked superiority of the HILIC/SALT approach compared to traditional HILIC and excellent complementarity to RPC for peptide separations. We believe these initial results offer a new dimension to HILIC, enabling it to transform from an occasional HPLC approach for peptide separations to a more generally applicable method.


Laparoscopic Versus Open Total Gastrectomy for Advanced Gastric Cancer: A Multicenter, Propensity Score-Matched Cohort Study in China.
Given the great technical difficulty and procedural complexity of laparoscopic total gastrectomy (LTG), the technical and oncologic safety of LTG versus open total gastrectomy (OTG) in the field of advanced gastric cancer (AGC) is yet undetermined. This multicenter cohort study aimed to compare the surgical and oncological outcomes of LTG with those of OTG in AGC patients. In total, 588 patients from 3 centers who underwent primary total gastrectomy with D2 lymphadenectomy, by well-trained surgeons with adequate experience, for pathologically confirmed locally AGC (T2N0-3, T3N0-3, or T4N0-3) between January 1, 2011, and December 31, 2015, were identified, and their clinical data were collected from three participating centers. After 1:1 propensity score matching (PSM), 450 cases (LTG, n = 225; OTG, n = 225) were eligible and assessed. No significant difference in the number of retrieved lymph nodes, 5-year disease-free survival (DFS) rates, or 5-year overall survival (OS) rates between both surgical groups were observed. Although LTG had significantly longer surgical time (262 vs. 180 min, p < 0.001), LTG was associated with fewer postoperative complications [relative risk (RR) 0.583, 95% CI 0.353-0.960, p = 0.047), less intraoperative bleeding (120 vs. 200 ml, p < 0.001), longer proximal margin resection (3 vs. 2 cm, p < 0.001), and shorter postoperative hospitalization (11 vs. 13 days, p < 0.001). The mortality rate was comparable in both groups. LTG was not inferior to OTG in terms of survival outcomes and was associated with shorter surgical and postoperative hospitalization time and fewer postoperative complications, suggesting LTG with D2 lymphadenectomy as an important alternative to OTG for patients with AGC, but to be carried out in highly experienced centers.


Long-term surgical outcomes of bilateral vs. unilateral medial rectus resection for recurrent exotropia.
The objective of this study is to compare the long-term outcomes of bilateral and unilateral medial rectus (BMR/UMR) resection for recurrent exotropia after bilateral lateral rectus (BLR) recession. Retrospective study was performed of 99 patients who underwent BMR resection (BMR group) or UMR resection (UMR group) for recurrent exotropia of 20-30 prism diopters (PD), with a minimum follow-up of 5 years. Surgical outcomes including success rate, exodrift rate, and average effect of MR resection were compared between two groups. The risk factors associated with poor outcomes were evaluated. At 5 years after surgery, 57% in the BMR group and 62% in the UMR group showed successful outcome. Success and recurrence rates were not significantly different between two groups, whereas the overcorrection rate was significantly higher in the BMR group (35% vs. 15%; p = 0.039). The average effect of MR resection was significantly greater after BMR throughout the whole postoperative period. The average effect of UMR resection was significantly greater in those who had previously undergone a large amount of BLR recession compared with those with a smaller dosage (p = 0.006). By multivariate analysis, a large amount of previous BLR recession and initial overcorrection of >10 PD of esotropia were found to be significant risk factors of overcorrection. In moderate angles of recurrent exotropia, large UMR resection is a safe and efficient procedure. However, if a large BLR recession was performed previously, surgical dosage for UMR resection should be reduced because of the high risk of long-term overcorrection.


Native seed preferences of shrub-steppe rodents, birds and ants: the relationships of seed attributes and seed use.
This study established the preferences of shrubsteppe granivores among seeds of 6 common sagebrushsteppe plants and related the preferences observed to physical and nutritional attributes of the seeds. Seeds of big sagebrush (Artemisia tridentata), cheatgrass (Bromus tectorum), Indian ricegrass (Oryzopsis hymenoides), western wheatgrass (Pascopyrum smithii), bitterbrush (Purshia tridentata) and green needlegrass (Stipa viridula) were placed in groups of petri dishes designed such that seed removal could be ascribed to either diurnal vertebrates, nocturnal vertebrates or ants. Though absolute quantities of seeds removed varied among the 3 granivore classes, calculations of preference based on weights of each seed species removed by each granivore class indicated that all 3 ranked the seeds similarly. Preference hierarchies of the 3 granivore classes were highly positively correlated with both calories per seed and % soluble carbohydrate of the seeds. The first correlation supports a basic prediction of optimal foraging theory -that foragers should maximize energy intake per unit time spent foraging. Both correlations emphasize the role of seed nutritional qualities in granivore seed selectivity in that soluble carbohydrate is a water-efficient energy source and its percentage is a good indicator of the digestible energy available in a food item. A corollary experiment comparing granivore use of an exotic seed (millet [Panicum miliaceum]) and a preferred native seed (Oryzopsis) demonstrated a distinct preference for the exotic. Since millet seeds are particularly high in % soluble carbohydrate, this result reinforced the apparent value of this nutritional attribute as a predictor of granivore seed preference. Among many seed resource characteristics upon which granivore seed selectivity might operate, our results indicate that individual species' nutritional composition may be particularly important. Thus, inferences about seed selectivity and resource partitioning among arid-land granivores should be interpreted with caution, especially those based on experiments using seed introductions, since the influence of seed nutritional attributes has not been widely acknowledged heretofore.


How does the addition of shape distinct gold nanoparticles influence on the conformational transition of poly(N-isopropylacrylamide)?
The subject of nanomaterials has created immense interest and expectations in the field of science and nanotechnology. Plentiful aspects proposed by gold nanoparticles (AuNPs) and their capability to affect macromolecular transition is the main driving force to execute the current study. A thermo-responsive polymer poly(N-isopropylacrylamide) (pNIPAM) is studied in presence of nanoparticles, particularly gold nanorods and nanospheres to elucidate completely the effect of their shape, surface area and structural morphology on the conformation of pNIPAM. In this respect, several biophysical techniques such as fluorescence spectroscopy, dynamic light scattering (DLS), Fourier transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC) are utilized to examine the interaction of Au nanospheres/nanorods with pNIPAM. For a better understanding of Au nanoparticles morphology, transmission electron microscopy (TEM) is also employed. Introducing gold nanoparticles with the polymeric solution promotes the polymer to stay in the coil conformation at a higher temperature than the LCST of aqueous pNIPAM. A shift of 2 and 25.5 °C in the LCST of pNIPAM is observed along with Au nanospheres and nanorods, respectively. The current study provides a better impact in the field of biomedical science specifically drug delivery and tissue engineering as the LCST approaches human body temperature.


Gamma knife radiosurgery for trigeminal neuralgia: comparing the use of a 4-mm versus concentric 4- and 8-mm collimators.
Gamma knife (GK) radiosurgery for trigeminal neuralgia (TN) has been effective in 50-80% of cases when using a single 4-mm collimator and a maximum dose of 60-90 Gy. Attempting to improve the response rate by increasing the dose may lead to increased risk of facial numbness or disturbed sensation. Combined use of 4- and 8-mm collimators results in a larger target volume, which would potentially treat a larger region of the nerve without increasing the maximum dose. One hundred-one patients suffering from medically refractory TN were evaluated. Fifty-four were treated with a single shot using a 4-mm helmet and 47 with concentrically aimed, equally weighted 4- and 8-mm helmets. 75 Gy were prescribed to the 100% isodose line (using a 4-mm helmet output factor of 0.80) in all cases. The outcome was assessed by interview or outpatient visit. An excellent/good response was seen in 57.8 vs. 71.4%, respectively, with a partial response of 13.3 vs. 2.3%, respectively (p > 0.05). Pain recurrence occurred in 28.6 vs. 23.2%, respectively (p > 0.05). The use of a combined concentric 4- and 8-mm collimator treatment for medically refractory TN at a maximum dose of 75 Gy does not improve outcome as compared with a single 4-mm collimator with an equivalent maximum dose.


Minichromosome maintenance protein 6, a proliferation marker superior to Ki-67 and independent predictor of survival in patients with mantle cell lymphoma.
Minichromosome maintenance protein 6 (MCM6) is one of six proteins of the MCM family which are involved in the initiation of DNA replication and thus represent a marker of proliferating cells. Since the level of cell proliferation is the most valuable predictor of survival in mantle cell lymphoma (MCL), we investigated lymph node biopsy specimens from 70 patients immunohistochemically with a monoclonal antibody against MCM6. The percentage of MCM6 expressing lymphoma cells ranged from 12.0 to 95.6%, with a mean of 61.0%, and was significantly higher than the percentage of Ki-67-positive cells (P<0.0001). Surprisingly, the ratio of MCM6-positive cells to Ki-67-positive cells was higher than in normal stimulated peripheral blood mononuclear cells, indicating a cell early G1-phase arrest in MCL. A high MCM6 expression level of more than 75% positive cells was associated with a significantly shorter overall survival time (16 months) compared to MCL with a low MCM6 expression level of less than 25% (no median reached, P<0.0001). Multivariate analysis revealed MCM6 to be an independent predictor of survival that is superior to the international prognostic factor and the Ki-67 index. Therefore, aside from gene expression profiling, immunohistochemical detection of MCM6 seems to be the most promising marker for predicting the outcome in MCL.


Sequential recognition of antigenic markers of Toxoplasma gondii tachyzoite by pooled sera of mice with experimental toxoplasmosis.
Accurate diagnosis of maternal toxoplasmosis can enhance the success of medical treatment and prevent congenital transmission. The current diagnostic methods have many limits, and they poorly differentiate between recent and latent infections. The present work was conducted to record the sequential recognition of antigenic markers of both Toxoplasma tachyzoites whole extract and glycosylinositolphospholipids (GIPLs)-enriched fraction by specific IgG and IgM, respectively, by immunoblotting analysis of the antigens against daily pooled serum samples from mice with experimentally induced recent and latent toxoplasmosis. IgG avidity immunoblotting was tested by using a wash with 6 M urea solution as antigen-antibody disrupting agent. Band of 10 kDa reacted exclusively with low-avidity IgG in pooled sera of mice with recent infection. Band of 39 kDa was a good marker for the infection; reacting with both low-avidity IgG in recent infection and with high-avidity IgG in latent one. Bands of 15, 23, 30, 60, 66, and 97 kDa reacted with variable avidity in both phases of infection. Two antigenic bands were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the GIPLs-enriched fraction of tachyzoite, the 14- and 30-kDa band. The 14-kDa band was recognized by IgM in pooled serum samples of recently infected mice only, while the 30-kDa band was recognized by serum samples of both recent and latent phases of infections. The study highlights the value of avidity immunoblotting assay to discriminate between recent and latent experimental toxoplasmosis. Further study must be carried on human to evaluate the values of the used technique.


Dual functionality of phosphonic-acid-appended phthalocyanines: inhibitors of urokinase plasminogen activator and anticancer photodynamic agents.
Phthalocyanines (Pcs) bearing phosphonic acid groups at the periphery exhibit a potential photodynamic effect to induce phototoxicity on human bladder cancer epithelial cells (UM-UC-3). In vitro photophysical and biological studies show high intrinsic ability to inhibit the activity of urokinase plasminogen activator (uPA) and matrix metalloproteinase-9 (MMP-9).


Paget's disease of the male breast: a case report.
The patient was a 91-year-old man with change in nipple appearance, itching and redness, and a palpable breast mass. At presentation, mammary Paget's disease (PD) was clinically suspected. Skin biopsy was performed and showed epidermis invaded by Paget cells, characterized by hyperchromatic nuclei and abundant pale-staining cytoplasm. Computed tomography and mammary ultrasonography confirmed the absence of an underlying invasive carcinoma, and the patient underwent right mastectomy and sentinel lymph node biopsy (SLNB). Both sentinel lymph nodes were found to be negative perioperatively, and further axillary dissection was not performed. Pathological results revealed no malignancy under the nipple, yet the Paget cells were more widely spread than expected. The patient was followed up without the need of postoperative chemotherapy. Male mammary PD is an extremely rare breast cancer, and there is no standard preoperative assessment or operative procedure. Mammography is many times unable to detect possible underlying breast carcinoma in female patients with mammary PD, and previous studies have reported that the detection rate was less than 50 %. However, some researchers reported that magnetic resonance imaging (MRI) might be more detectable to confirm the extent of the cancer. The extent of the skin change around the nipple is often different from the actual perimeter of Paget cells. In extra-mammary PD, mapping biopsy is known to be useful to determine areas free of cancer. The benefits of SLNB have also been demonstrated for the management of less invasive breast cancers, and previous reports have shown that the use of SLNB is reasonable for treatment of mammary PD without underlying invasive cancer. MRI, mapping biopsy, and SLNB are all less invasive procedures and thus may be suitable for treatment of male mammary PD.


A neural mechanism underlying predictive visual motion processing in patients with schizophrenia.
Psychotic symptoms may be traced back to sensory sensitivity. Thereby, visual motion (VM) processing particularly has been suggested to be impaired in schizophrenia (SCZ). In healthy brains, VM underlies predictive processing within hierarchically structured systems. However, less is known about predictive VM processing in SCZ. Therefore, we performed fMRI during a VM paradigm with three conditions of varying predictability, i.e., Predictable-, Random-, and Arbitrary motion. The study sample comprised 17 SCZ patients and 23 healthy controls. We calculated general linear model (GLM) analysis to assess group differences in VM processing across motion conditions. Here, we identified significantly lower activity in right temporoparietal junction (TPJ) for SCZ patients. Therefore, right TPJ was set as seed for connectivity analyses. For patients, across conditions we identified increased connections to higher regions, namely medial prefrontal cortex, or paracingulate gyrus. Healthy subjects activated sensory regions as area V5, or superior parietal lobule. Reduced TPJ activity may reflect both a failure in the bottom-up flow of visual information and a decrease of signal processing as consequence of increased top-down input from frontal areas. In sum, these altered neural patterns provide a framework for future studies focusing on predictive VM processing to identify potential biomarkers of psychosis.


Thymoquinone administration ameliorates Alzheimer's disease-like phenotype by promoting cell survival in the hippocampus of amyloid beta1-42 infused rat model.
Thymoquinone (TQ), a biologically active ingredient of Nigella sativa, has anti-inflammatory, anti-oxidative and neuroprotective properties. Therefore, it could be a good candidate in the recovery of Alzheimer`s disease (AD) pathology rather than current symptomatic reliefs. In the present study, we examined the molecular healing effects of TQ in amyloid beta 1-42 (Aβ1-42) peptide-infused AD rat hippocampus. A micro-osmotic pump containing aggregated Aβ1-42 was cannulated into the hippocampus of adult female rats. After two weeks infusion, the dose of TQ (10 mg/kg or 20 mg/kg) was determined according to the HPLC results of cerebrospinal fluid and TQ was given to rats intragastrically for 15 days. The memory performance of rats was determined by Morris water maze test. Afterwards, the acetylcholinesterase (AChE) level were measured by ELISA. Histopathological examinations of hippocampal tissue were performed for cell survival by Nissl staining, for detection of amyloid plaque deposits by Congo red staining and for determination of degenerating neurons by Fluoro Jade C staining. MicroRNA/mRNA levels and protein expressions of AD-related genes and proteins were analyzed by Real-Time Polymerase Chain Reaction and Western Blotting, respectively. Administration of TQ enhanced the memory performance of Aβ1-42 infused rats and it also ameliorated the neuronal loss in the cornu ammonis (CA1), but not in the dentate gyrus (DG). In addition, TQ treatment decreased the fibril deposition whose accumulation was significantly higher in the Aβ1-42-infused animals compared to that of the control group. The expression profiles of mir29c and Bax which significantly upregulated in the Aβ1-42-infused animals were attenuated by TQ. Furthermore, administration of TQ decreased the expressions of Aβ, phosphorylated-tau, and BACE-1 proteins. There was no significant therapeutic effect of TQ on the AKT/GSK3β or MAPK signaling pathways which were affected due to Aβ1-42 infusion. TQ has the capacity to recover the neuropathology by removing Aβ plaques and by restoring neuron viability. All might have established the molecular basement of the consolidation in the memory observed by means of TQ treatment.


Controllability and duration of stress alter central nervous system depressant-induced sleep time in rats.
Rats were exposed to either 80 escapable shocks or yoked inescapable shocks and then injected with several hypnotic doses of sodium pentobarbital, midazolam, or ethanol; their sleep-time duration was compared with that of naive controls. Inescapable shock exposure resulted in a significant increase in ethanol-induced sleep time compared with the escapable shock and naive control groups. Both escape and yoked groups showed an increase in barbiturate-induced sleep time compared with controls, although no difference was observed for midazolam. Acute stress (twenty 5-s inescapable shocks) did not alter the depressant-induced sleep time for any of the drugs tested. These results illustrate the importance of psychological aspects of stress and its influence on the potency of certain depressants.


Anatomy-Based Frequency Allocation in Cochlear Implantation: The Importance of Cochlear Coverage.
This study aimed to compare the predicted anatomy-based frequency allocation of cochlear implant electrodes with the default standard frequencies. Retrospective study. A retrospective analysis was performed using computed tomography (CT) images of patients who received cochlear implants at a tertiary referral center. Patients were excluded if they had any congenital or acquired cochlear anatomical anomalies. The CT images of the patients were uploaded to the surgical planning software. Two independent reviewers allocated the anatomical parameters of the cochlea. The software then used these parameters to calculate the frequency allocation for each electrode according to the type of electrode and the length of the organ of Corti (OC) in each patient. These anatomy-based frequency allocations were compared with the default frequency settings. Frequency-to-place mismatch in semitones. A total of 169 implanted ears in 102 patients were included in this study. The readings of the two reviewers were homogenous, with a Cronbach's alpha of 0.98. The mean anatomy-based frequency allocation was 487.3 ± 202.9 Hz in electrode 1; 9,298.6 ± 490.6 Hz in electrode 12. The anatomy-based frequency allocations were found to be significantly higher than the frequencies of the default frequencies for each corresponding electrode (one-sample t-test, P < .001). The frequency-to-place mismatch was negatively correlated with cochlear coverage and positively correlated with the cochlear duct length (Pearson correlation > 0.65, P < .003). The anatomy-based frequency allocation of each electrode is significantly different from the default frequency setting. This frequency-to-place mismatch was affected mainly by the cochlear coverage. 3 Laryngoscope, 132:2224-2231, 2022.


Separation of porphyrin isomers by high-performance liquid chromatography.
A reversed-phase gradient elution system is described for the simultaneous separation of the type I and type III isomers of 8-, 7-, 6-, 5- and 4-carboxylated porphyrins and isocoproporphyrins. The method, adaptable for isocratic and stepwise separation of individual groups of isomers, is also suitable for preparative isolation of pure porphyrins. The analyses of porphyrin isomers in the urine and faeces of porphyric patients are examples of applications.


Risk Factors for Incident Diabetes in a Cohort Taking First-Line Nonnucleoside Reverse Transcriptase Inhibitor-Based Antiretroviral Therapy.
Efavirenz is the preferred nonnucleoside reverse transcriptase inhibitor (NNRTI) in first-line antiretroviral therapy (ART) regimens in low- and middle-income countries, where the prevalence of diabetes is increasing. Randomized control trials have shown mild increases in plasma glucose in participants in the efavirenz arms, but no association has been reported with overt diabetes. We explored the association between efavirenz exposure and incident diabetes in a large Southern African cohort commencing NNRTI-based first-line ART. Our cohort included HIV-infected adults starting NNRTI-based ART in a private sector HIV disease management program from January 2002 to December 2011. Incident diabetes was identified by the initiation of diabetes treatment. Patients with prevalent diabetes were excluded. We included 56,298 patients with 113,297 patient-years of follow-up (PYFU) on first-line ART. The crude incidence of diabetes was 13.24 per 1000 PYFU. Treatment with efavirenz rather than nevirapine was associated with increased risk of developing diabetes (hazard ratio 1.27 (95% confidence interval (CI): 1.10-1.46)) in a multivariate analysis adjusting for age, sex, body mass index, baseline CD4 count, viral load, NRTI backbone, and exposure to other diabetogenic medicines. Zidovudine and stavudine exposure were also associated with an increased risk of developing diabetes. We found that treatment with efavirenz, as well as stavudine and zidovudine, increased the risk of incident diabetes. Interventions to detect and prevent diabetes should be implemented in ART programs, and use of antiretrovirals with lower risk of metabolic complications should be encouraged.


Predictors and outcomes following naloxone administration during Phase I anesthesia recovery.
To identify characteristics associated with postoperative respiratory depression that required naloxone intervention during Phase I recovery following general anesthesia. A secondary aim is to compare postoperative outcomes between patients who received naloxone and those who did not. Patients who received naloxone to reverse opioid-induced respiratory depression or sedation during Phase I postanesthesia recovery from January 1, 2010 to December 31, 2013 were identified and matched to 2 controls based on age, sex, and surgical procedure during the same year. A chart review was performed to identify factors associated with risk for intervention requiring naloxone as well as to note the occurrence of adverse postoperative outcomes. Analyses to assess characteristics potentially associated with naloxone use were performed using conditional logistic regression taking into account the 1:2 matched set case-control study design. Naloxone was administered to 413 patients, with an incidence of 2.5 per 1000 anesthetics [95 % confidence interval (CI) 0.7-6.5]. Presence of obstructive sleep apnea [odds ratio (OR) = 1.74, 95 % CI 1.22-2.48, P = 0.002], ASA Physical Status (PS) ≥III (OR 1.44, 95 % CI 1.08-1.92, P = 0.013), and greater opioid administration (OR 1.22, 95 % CI 1.12-1.33, per 10 intravenous morphine equivalents mg, P < 0.001) were associated with naloxone administration. Naloxone administration was associated with increased adverse events (OR 3.39, 95 % CI 2.22-5.23, P < 0.001). Obstructive sleep apnea, higher ASA-PS scores and greater doses of intraoperative opioids were associated with naloxone administration during Phase I recovery. Patients administered naloxone had increased adverse events after discharge from the recovery room and may benefit from a higher level of postoperative care.


Clues to pathogenesis of fibromyalgia in patients with sickle cell disease.
To investigate the association between sickle cell disease (SCD) and fibromyalgia (FM). Nine patients with SCD for whom a rheumatology consult was requested were assessed for FM by retrospective chart review. Eleven inpatients with other forms of anemia referred for rheumatology consult were also assessed for FM. Eight of 9 patients with SCD fulfilled classification criteria for FM compared to one of 11 patients without SCD (p < 0.001). Awareness of the high frequency of FM in SCD can improve treatment of sickle cell crisis. Some pain that is labeled as sickle cell crisis pain may be due to FM, and may improve with tender point injections.


P16(INK4A) is implicated in both the immediate and adaptative response of human keratinocytes to UVB irradiation.
The p16(INK4A-ARF) locus plays a crucial role in the control of cellular proliferation via both the Rb and P53 pathways. We previously demonstrated that this locus is altered in human skin carcinomas. In the present study we have studied the expression of the p16(INK4A-ARF) locus following UVB irradiation of normal human keratinocytes both at the mRNA (RT-PCR) and at the protein (Western blotting) levels. Our data confirmed that P16(INK4A) protein is induced by UVB at low (30 mJ cm(2)) and high (100 mJ cm(2)) doses and is observed after a single or repeated exposure implying that this response is involved in both the immediate and adaptative response to UVB. The apparent absence of induction p16(INK4A) mRNA suggested that P16(INK4A) protein is upregulated at the post-transcriptional level. Analysis by flow cytometry and BrdU staining indicated that the highest protein level of P16(INK4A) in the cells was associated with a G(2) cell cycle arrest. Comparative analysis of P16(INK4A) and P53 showed that they were differentially modulated in keratinocytes according to the UVB dose and regimen. Low, acute or repeated UVB exposures led to accumulation of both P16(INK4A) and p53, whereas at high UVB doses, P53 and P53-dependent genes were not induced or even downregulated and only a slight but reproducible stabilization of P16(INK4A) protein was observed. In our conditions, P14(ARF) did not seem to participate in the UV response in these cells as P14(ARF) protein did not vary. These results infer that P16(INK4A) plays a role in cell cycle regulation of keratinocytes submitted to UVB irradiation. They also reinforce our previous demonstration of the importance of inactivation of this gene in UV-induced skin carcinogenesis.


High cell density perfusion culture of hybridoma cells recycling high molecular weight components.
We have developed a high cell density and high product concentration culture system recycling high molecular weight components. The production of monoclonal antibodies in high concentration was performed by this culture system with mouse human hybridoma H2 and V6 cells in serum-free defined media.The concentration of IgG after 48 days culture of H2 cells in ITES-eRDF reached 2 mg/ml and the purity of IgG in culture fluid was 61%. In addition, high molecular weight components in serum-free media, such as transferrin or BSA, could be reduced to 5% of the original concentration.


Protective effect of carnosine during nitrosative stress in astroglial cell cultures.
Formation of nitric oxide by astrocytes has been suggested to contribute, via impairment of mitochondrial function, to the neurodegenerative process. Mitochondria under oxidative stress are thought to play a key role in various neurodegenerative disorders; therefore protection by antioxidants against oxidative stress to mitochondria may prove to be beneficial in delaying the onset or progression of these diseases. Carnosine has been recently proposed to act as antioxidant in vivo. In the present study, we demonstrate its neuroprotective effect in astrocytes exposed to LPS- and INFgamma-induced nitrosative stress. Carnosine protected against nitric oxide-induced impairment of mitochondrial function. This effect was associated with decreased formation of oxidatively modified proteins and with decreased up-regulation oxidative stress-responsive genes, such as Hsp32, Hsp70 and mt-SOD. Our results sustain the possibility that carnosine might have anti-ageing effects to brain cells under pathophysiological conditions leading to degenerative damage, such as aging and neurodegenerative disorders.


From Crisis to Insight: Navigating Public Health Challenges in an Aging Society-Lessons from Hong Kong's COVID-19 Experience.
This article critically examines the experience of Hong Kong (HK) with the COVID-19 pandemic and highlights the urgent need for effective public health responses in an aging society. As one of the world's fastest aging societies, Hong Kong's vulnerability to the consequences of pandemics is particularly pronounced due to low birth rates and high life expectancies. The city has experienced multiple waves of the COVID-19 virus, with the fifth wave causing some of the highest mortality rates in the world. However, through continuous learning and adaptation, HK's public health response has evolved to combat the crisis effectively. The discussion aims to highlight the importance of drawing from both positive and negative lessons from past epidemics and the critical role of public health systems in ensuring the well-being of older adults and the wider community. It emphasizes the need for sustained efforts to maintain and strengthen public health systems to tackle the ongoing and future public health challenges in an aging society.


Diagnostic and mechanistic values of microRNA-130a and microRNA-203 in patients with papillary thyroid carcinoma.
This research was determined to unearth the diagnostic values and the effects of microRNA (miR)-130a and miR-203 on cell proliferation and apoptosis of papillary thyroid carcinoma (PTC). Expression of miR-130a and miR-203 were evaluated and were subjected to correlation analysis. The diagnostic values of miR-130a and miR-203 and their associations with clinicopathological characteristics of patients with PTC were measured. The expression levels of miR-130a and miR-203 in K1, IHH4, TPC-1, and BCPAP cells together with Nthy-ori 3-1 cells were measured. Cells were transfected with miR-130a mimics, miR-203 mimics, and coordinate of miR-130a mimics and miR-203 mimics. Cell growth, colony formation, and apoptosis were detected by cell counting kit-8 (CCK-8) assay, colony formation assay, and flow cytometry. PTC tissues had decreased miR-130a and miR-203 relative to adjacent normal tissues and normal thyroid tissue (both P < .05). miR-130a was in positive correlation with miR-203 (r = 0.754, P < .01). miR-130a was related with tumor infiltration and tumor stage while miR-203 was implicated in tumor stage and lymph-node metastasis. The area under the curve (AUC), sensitivity, as well as specificity for miR-130 in predicting PTC was 0.839, 74.5%, and 85.0% and those for miR-203 were 0.818, 73.7%, and 84.0%, respectively. PTC cells had lower expression of miR-130a and miR-203 than that in Nthy-ori 3-1 cells. After transfected miR-130a and miR-203 mimics in BCPAP and TPC-1 cells, both cells had increased miR-130a and miR-203, promoted cell apoptosis rate and decreased cell growth rate, and colony formation ability. After coordinately transfected with miR-130a mimics and miR-203 mimics, the cell growth and colony formation ability of PTC cells were restrained, and apoptosis of PTC cells was elevated (all P < .05). This study highlights that miR-130a and miR-203 have satisfactory diagnostic value in PTC and upregulated miR-130a and miR-203 can inhibit PTC cell growth and promote cell apoptosis.


Positive and Negative Control of Enhancer-Promoter Interactions by Other DNA Loops Generates Specificity and Tunability.
Enhancers are ubiquitous and critical gene-regulatory elements. However, quantitative understanding of the role of DNA looping in the regulation of enhancer action and specificity is limited. We used the Escherichia coli NtrC enhancer-σ54 promoter system as an in vivo model, finding that NtrC activation is highly sensitive to the enhancer-promoter (E-P) distance in the 300-6,000 bp range. DNA loops formed by Lac repressor were able to strongly regulate enhancer action either positively or negatively, recapitulating promoter targeting and insulation. A single LacI loop combining targeting and insulation produced a strong shift in specificity for enhancer choice between two σ54 promoters. A combined kinetic-thermodynamic model was used to quantify the effect of DNA-looping interactions on promoter activity and revealed that sensitivity to E-P distance and to control by other loops is itself dependent on enhancer and promoter parameters that may be subject to regulation.


Precentral and inferior prefrontal hypoactivation during facial emotion recognition in patients with schizophrenia: A functional near-infrared spectroscopy study.
Although patients with schizophrenia demonstrate abnormal processing of emotional face recognition, the neural substrates underlying this process remain unclear. We previously showed abnormal fronto-temporal function during facial expression of emotions, and cognitive inhibition in patients with schizophrenia using functional near-infrared spectroscopy (fNIRS). The aim of the current study was to use fNIRS to identify which brain regions involved in recognizing emotional faces are impaired in patients with schizophrenia, and to determine the neural substrates underlying the response to emotional facial expressions per se, and to facial expressions with cognitive inhibition. We recruited 19 patients with schizophrenia and 19 healthy controls, statistically matched on age, sex, and premorbid IQ. Brain function was measured by fNIRS during emotional face assessment and face identification tasks. Patients with schizophrenia showed lower activation of the right precentral and inferior frontal areas during the emotional face task compared to controls. Further, patients with schizophrenia were slower and less accurate in completing tasks compared to healthy participants. Decreasing performance was associated with increasing severity of the disease. Our present and prior studies suggest that the impaired behavioral performance in schizophrenia is associated with different mechanisms for processing emotional facial expressions versus facial expressions combined with cognitive inhibition.


Efficacy of perioperative synbiotics treatment for the prevention of surgical site infection after laparoscopic colorectal surgery: a randomized controlled trial.
The aim of this study was to assess the effect of perioperative oral administration of synbiotics on the surgical outcome in patients undergoing laparoscopic colorectal resection. In this single-center randomized, controlled trial, patients scheduled to undergo elective laparoscopic colorectal surgery were eligible to participate and randomly assigned to a synbiotics group or a control group. The primary study outcome was the development of infectious complications, particularly surgical site infection (SSI), within 30 days of surgery. In this study, 379 patients were enrolled and randomly assigned (173 to the synbiotics group and 206 to the control group), of whom 362 patients (168 to the synbiotics group and 194 to the control group) were eligible for this study. SSI occurred in 29 (17.3%) patients in the synbiotics group and 44 (22.7%) patients in the control group (OR: 0.761, 95% CI 0.50-1.16; p = 0.20). Overall, the rate of postoperative complications, including anastomotic leakage, did not differ significantly between the two groups. Synbiotics treatment reversed the changes in fecal bacteria and organic acids after surgery and suppressed the increases in potentially pathogenic species, such as Clostridium difficile. The efficacy of perioperative administration of synbiotics was not validated as a treatment for reducing the incidence of infectious complications after laparoscopic colorectal resection. However, the microbial imbalance, in addition to the reduction in organic acids, could be improved by perioperative synbiotics treatment.


Homologous sequences in steroidogenic enzymes, steroid receptors and a steroid binding protein suggest a consensus steroid-binding sequence.
The amino acid sequences of two steroidogenic enzymes, P450c17 (steroid 17 alpha-hydroxylase/17,20 lyse) and P450c21 (steroid 21-hydroxylase), are only 28.9% identical. However, these proteins share a region of 21 amino acids bearing 17 identical residues, which we previously suggested may represent the steroid binding site. We assembled a sequence database of known steroid-binding proteins and searched this with the sequence of this 21 amino acid region. The steroidogenic enzymes, P450c17, P450c21, P450scc (the cholesterol side-chain cleavage enzyme), and P450c11 (steroid 11 beta/18-hydroxylase) share a subregion of 17 amino acids having at least 15 identical residues. Related sequences were identified in a computerized search of the available sequences of steroid hormone receptors and binding proteins. These sequences were invariably found within larger domains previously associated with steroid binding. From these we propose a more general consensus sequence of LPLLL +/- 000KDRE0LKRL +/- PV, where +/- refers to any charged amino acid, and 0 refers to an uncharged amino acid. This consensus sequence predicts 147 or 187 total amino acids in 11 human proteins examined (78.6%). An equivalent degree of sequence identity, 178 of 221 amino acids (80.5%) was found among 13 animal homologs of these human proteins. The ability of this consensus sequence to predict 325 of 408 amino acids (79.7%) strongly suggests this sequence is necessary, if not sufficient, for a steroid binding site in many proteins. Lecithin-cholesterol acetyl transferase, cholesterol ester transfer protein, and steroid sulfatase did not have sequences similar to our consensus sequence.


Anaesthetic risks in osteoarticular disorders.
Anaesthesia in patients with either treated or untreated osteo-articular disorders poses a number of specific problems. The authors deal with risks resulting from mechanical deformities produced by these disorders, by the involvement of a large number of functional systems, by changes in the immune status of such patients and by alterations in the reactions of arthritic patients resulting from prior or concurrent treatments. The authors emphasize the crucial points which require evaluation and observation before and during anaesthesia.


Effect of fixatives on rat brain acetylcholinesterase activity.
Biochemical measurement of acetylcholinesterase (AChE) activity on sonicates of samples of rat cortex, striatum and cerebellum which had been treated with either 4% formaldehyde (HCHO) or 4% HCHO/0.5% glutaraldehyde suggest that fixation of tissue causes a rather large loss (50-80%) in AChE activity which may vary considerably from region to region of brain. Attempts to protect the activity by treatment with diisopropyl fluorophosphate (DFP) before fixation and reactivation after fixation with pyridine-2-aldoxime methiodide (PAM) were unsuccessful. The results suggest care must be exercised in attempting to use AChE histochemistry on fixed tissue for quantitative measurements.


EUS-guided Trucut mural biopsies in the investigation of unexplained thickening of the esophagogastric wall.
In patients with thickened esophagogastric wall on CT but without evidence of malignancy on endoscopic mucosal biopsies, obtaining adequate histology from the gut wall is difficult. We examined whether it is feasible to obtain diagnostic tissue core from the gut wall by using EUS-guided Trucut biopsy technique in this group of patients. Ten patients were included in this study. Under EUS guidance, mucosa was penetrated by using a 19-gauge Trucut needle, and a 18-mm tissue tray was advanced obliquely through the wall layers to avoid penetration of the serosa. Then, the cutting sheath was fired over the tray. Biopsies were performed without complications. Diagnoses of carcinoma were made in 5 patients. Four other patients had benign histology, and, during follow-up, all these diagnoses have been proven to be true negatives. One Trucut biopsy specimen was considered false negative. The EUS-guided Trucut mural biopsy technique could yield diagnostic tissue cores in patients with unexplained thickening of the esophagogastric wall.


Effectiveness and tolerability of mirtazapine and amitriptyline in alcoholic patients with co-morbid depressive disorder: a randomized, double-blind study.
Studies indicate that serotoninergic and noradrenergic pathophysiological mechanisms may underlie both alcohol abuse/dependence and depressive disorder. The purpose of this study was to evaluate and compare the effectiveness and tolerability of two serotonergic and noradrenergic antidepressant drugs-mirtazapine and amitriptyline, for the treatment of patients with alcohol dependence co-morbid with depressive disorder in a randomized, double-blind treatment setting. Forty-four patients were included in the study. Twenty-four patients were randomized to mirtazapine and twenty to amitriptyline groups. Thirty-six of them completed the study. The 17-item Hamilton Depression Rating Scale (HDRS), the Spielberger State-Trait Anxiety Inventory (STAI) and alcohol craving questionnaire were used at baseline and, at days 7, 14, 28, 42, and 56 to estimate the effectiveness of the antidepressant treatment. Michigan Alcoholism Screening Test (MAST) was used in the assessment of alcohol dependence. The tolerability was assessed with the Udvalg for Kliniske Undersogelser Side Effect Rating Scale (UKU). There was significant improvement in HDRS and alcohol craving scores with both the drugs. However there were no statistical differences between treatment groups. Mirtazapine was tolerated better than amitriptyline treatment. The treatment with either mirtazapine or amitriptyline resulted with the reduction of HDRS and craving scores. The side-effect profile of mirtazapine was relatively favorable in our study.


Optimization of method for achieving a single-cell suspension from mouse corneas.
The single-cell RNA-sequencing (scRNA-seq) technique is used to explore the biological characteristics of tissues under pathological and physiological conditions that include certain chronic eye diseases. Harvesting of single-cell suspensions is one challenge inherent to scRNA-seq procedures. This study aimed to use an optimized method to digest a whole mouse cornea to harvest single-cell suspensions. We utilized five different mouse cornea digestion methods to obtain single-cell suspensions: (1) 5 dissected mouse corneas were cut into pieces (∼0.5 mm) and digested in trypsin for 10 min, and this digestion was repeated for 10 cycles; (2) 5 dissected mouse corneas were cut into pieces and incubated with 5 mg/ml collagenase A at 37 °C for 1h and then further digested in trypsin at 37 °C for 10 min; (3) used the same approach as that used in method 2, but the second digestion step was performed in TrypLE for 20 min; (4) used the same approach as that used in method 2, but the concentration of collagenase A was 2 mg/ml and the incubation time was 2h; (5) used the same approach as that used in method 3, but the corneas were incubated in 2 mg/ml collagenase A at 37 °C for 2h. Trypan blue staining was used to calculate the cell viability and agglomeration rate. The cell types and percentages were determined using immunofluorescence staining. RNA integrity number (RIN) was measured by Agilent 2100. Method 1 showed the lowest cell yield (0.375 × 105), epithelial cell percentage, and less than 70% cell viability, thus not a proper protocol. Method 2 showed the highest cell viability (over 90%), percentage of single-cell (89.53%), and high cell quantity (1.05 × 105). Method 3 had a significantly lower cell viability (55.30%). Cell agglomeration rates of method 4 and 5 reached up to 20% and 13%, and with lower cell viability (72.51%, 59.87%, respectively) and decreased epithelial cell rate compared to method 2 and 3. The results suggest that method 2 (5 mg/ml collagenase A and trypsin) is a preferred protocol for digesting mouse cornea to obtain single-cell suspension which achieves the criterion of single-cell RNA sequencing.


Genotyping by mass spectrometric analysis of short DNA fragments.
A method has been developed to produce small DNA fragments from PCR products for analysis of defined DNA variations by mass spectrometry. The genomic region to be analyzed is PCR-amplified with primers containing a sequence for the type IIS restriction endonuclease Bpml. Bpml digestion of the resultant PCR products yields fragments as small as seven bases, which are then analyzed by electrospray ionization mass spectrometry. The approach was validated using seven different variants within the APC tumor suppressor gene, in which a perfect correlation was obtained with DNA sequencing. Both the sense and antisense strands were analyzed independently, and several variants can be analyzed simultaneously. These results provide the basis for a generally applicable and highly accurate method that directly queries the mass of variant DNA sequences.


Effect of body shape and inflammation on tooth loss in men and women.
Android and gynoid adiposity is differently involved on inflammatory responses in men or women in periodontitis and tooth loss. We tested the hypothesis whether identical waist-to-hip ratios (WHR) in men and women could abolish this disparity. Data of 2746 participants from the Study of Health in Pomerania (SHIP) were analysed. Men and women were 1:1 matched, N = 344:344, in three age strata for waist-to-hip ratio. We determined anthropometric measures, attachment loss, tooth loss and markers of systemic inflammation. Women matched with men by WHR had increased periodontal measures as compared to women of the general population. Nevertheless, in the matched pairs incidence risk (IRR) ratios for any tooth loss associated with elevated C-reactive protein were IRR = 2.15 (CI 1.33-3.40) and 1.04 (0.66-1.66) in men and women, respectively. Regression with tooth loss due to any cause as dependent variable showed dose dependency on C-reactive protein levels in men but not in women. The adjusted IRR associated with high C-reactive protein in men was 1.37 (CI 1.05-1.78) and 2.63 (1.58-4.38) in general and in matched subjects, respectively. Systemic inflammation was associated with tooth loss in men but not in women even in women with wide girth. Despite worsened periodontal measures and inflammation in women matched for body shape with men, these women do not lose more teeth even when they are exposed to increased markers of systemic inflammation. This is an attempt to disentangle the unclear relationship between obesity and periodontitis, both of them having public health relevance.


[Sjögren-Larsson syndrome: a novel mutation in a Moroccan child].
Sjögren-Larsson syndrome is a very rare inherited neurocutaneous disorder caused by a deficiency of microsomal enzyme fatty aldehyde dehydrogenase (FALDH). The authors report a case of typical Sjögren-Larsson syndrome in a 7-year-old Moroccan child who presented with classical symptoms (congenital ichthyosis, mental retardation, and spastic paraparesis) and epilepsy. The genetic study revealed a new mutation in the FALDH gene mapped to chromosome 17, consisting in a G109A substitution in exon 2.


Gastro-oesophageal reflux in children: comparison of different durations, positions and sleep-awake periods of pH monitoring in the same patient.
A group of 72 children (mean age: 21.7 months, range: 14 days-19 years) with symptoms of gastro-oesophageal reflux were investigated by 22 h pH monitoring. Using a Proxeda software, we compared, in the same patient, the specificity and sensitivity of pH monitoring during 3 h, 6 h, 12 h, 12 nocturnal hours and 3 postprandial hours, as well as the influence of position and the sleep and alert periods. Results showed that all the short pH monitorings were statistically less sensitive than 22 h pH monitoring (P < 0.025). As regards specificity, only the 12 nocturnal hours pH monitoring was not statistically different from the 22 h pH monitoring. Gastro-oesophageal reflux was more frequent when the patient was awake than during sleep. pH monitoring seemed more reliable in the recumbent than in the upright position. We conclude that long-term pH monitoring (22 h) is the test of choice to diagnose gastro-oesophageal reflux because it included sleep and alert periods as well as different positions.


Induction of the proteolytic activity of a membrane protein in Plasmodium falciparum by phosphatidyl inositol-specific phospholipase C.
Membrane anchoring of proteins by a covalently attached glycosyl-phosphatidylinositol moiety has been reported in many different eukaryotic cells including parasite protozoa. The diversity of proteins in which this phospholipid attachment is found suggests that it is functionally important and perhaps also functionally pleiotropic. Studies on the Thy-1 antigen of murine lymphocytes indicate that it can facilitate the lateral mobility of membrane proteins. It can also permit the rapid and specific release of the anchored proteins from the membrane following cleavage by a phosphatidylinositol-specific phospholipase C (PI-PLC). Here we show that this type of anchoring may be involved in the regulation of an enzymatic activity. PI-PLC releases a Plasmodium falciparum membrane protein of relative molecular mass (Mr) 76K (p76) from intact merozoites or isolated schizont membranes and induces a proteolytic activity associated with its soluble form. Endogenous activation of the proteolytic activity of p76 appears to occur at the end of the schizogony and could initiate a cascade of biochemical events associated with merozoite maturation.


Self-assembling DNA hydrogel-based delivery of immunoinhibitory nucleic acids to immune cells.
Immunoinhibitory oligodeoxynucleotides (INH-ODNs) are promising inhibitors of Toll-like receptor 9 (TLR9) activation. To efficiently deliver INH-ODNs to TLR9-positive cells, we designed a Takumi-shaped DNA (Takumi) consisting of two partially complementary ODNs as the main component of a DNA hydrogel. Polyacrylamide gel electrophoresis showed that Takumi-containing INH-ODNs (iTakumi) and iTakumi-based DNA hydrogel (iTakumiGel) were successfully generated. Their activity was examined in murine macrophage-like RAW264.7 cells and DC2.4 dendritic cells by measuring tumor necrosis factor-α and interleukin-6 release after the addition of a TLR9 ligand (CpG ODN). Cytokine release was efficiently inhibited by the iTakumiGel. Flow cytometry analysis and confocal microscopy showed that cellular uptake of INH-ODN was greatly increased by the iTakumiGel. These results indicate that a Takumi-based DNA hydrogel is useful for the delivery of INH-ODNs to immune cells to inhibit TLR9-mediated hyperinduction of proinflammatory cytokines. From the Clinical Editor: Toll-like receptor 9 activation has been reported to be associated with many autoimmune diseases. DNA inhibition using oligodeoxynucleotides is one of the potential treatments. In this article, the authors described hydrogel-based platform for the delivery of the inhibitory oligodeoxynucleotides for enhanced efficacy. The positive findings could indicate a way for the future.


Personalized redox biology: Designs and concepts.
Personalized interventions are regarded as a next-generation approach in almost all fields of biomedicine, such as clinical medicine, exercise, nutrition and pharmacology. At the same time, an increasing body of evidence indicates that redox processes regulate, at least in part, multiple aspects of human physiology and pathology. As a result, the idea of applying personalized redox treatments to improve their efficacy has gained popularity among researchers in recent years. The aim of the present primer-style review was to highlight some crucial yet underappreciated methodological, statistical, and interpretative concepts within the redox biology literature, while also providing a physiology-oriented perspective on personalized redox biology. The topics addressed are: (i) the critical issue of investigating the potential existence of inter-individual variability; (ii) the importance of distinguishing a genuine and consistent response of a subject from a chance finding; (iii) the challenge of accurately quantifying the effect of a redox treatment when dealing with 'extreme' groups due to mathematical coupling and regression to the mean; and (iv) research designs and analyses that have been implemented in other fields, and can be reframed and exploited in a redox biology context.


Effects of calcium oxalate monohydrate crystals on expression and function of tight junction of renal tubular epithelial cells.
Tight junction has a crucial role in regulating paracellular transports (as a barrier) and in separating apical from basolateral compartments to maintain cell polarity (as a fence). Tight junction can be disrupted by various stimuli, including oxidative stress, pathogens and proinflammatory cytokines. However, association of defective tight junction with kidney stone pathogenesis remains unknown. We therefore examined whether calcium oxalate monohydrate (COM) crystals, which are the major crystalline composition in kidney stones, have any effects on expression and function of tight junction of polarized renal tubular epithelial cells. Western blot analysis revealed marked decrease in levels of occludin and zonula occludens-1 (ZO-1) in COM-treated polarized Madin-Darby canine kidney (MDCK) cells. Immunofluorescence staining revealed not only the decline of these tight junction proteins but also their redistribution and dissociation in COM-treated cells. Additionally, transepithelial resistance was significantly decreased, indicating impaired tight junction barrier and increased paracellular permeability in COM-treated cells. Subcellular fractionation followed by western blot analysis of Na(+)/K(+)-ATPase-α1 revealed that this basolateral membrane marker was also detectable in apical membrane fraction of COM-treated cells, but not in apical membrane fraction of control cells. Immunofluorescence study confirmed the translocation of Na(+)/K(+)-ATPase-α1 (from basolateral to apical membranes) in COM-treated cells, indicating impaired fence function of the tight junction. Moreover, dihydrorhodamine assay using flow cytometry revealed the significantly higher level of hydrogen peroxide in the COM-treated cells. These data provide the first evidence to demonstrate decreased expression and defective barrier and fence functions of the tight junction of renal tubular epithelial cells exposed to COM crystals that may be fundamental for subsequent renal tubulointerstitial injury, which in turn enhances the stone pathogenesis.


Higher order structure in the 3'-minor domain of small subunit ribosomal RNAs from a gram negative bacterium, a gram positive bacterium and a eukaryote.
An experimental approach was used to determine and compare the highest order structure within the 150 to 200 nucleotides at the 3'-ends of the RNAs from the small ribosomal subunits of Escherichia coli, Bacillus stearothermophilus and Saccharomyces cerevisiae. Chemical reagents were employed to establish the degree of stacking and/or accessibility of each adenosine, guanosine and cytidine. The double helices were probed with a cobra venom ribonuclease from Naja naja oxiana, and the relatively unstructured and accessible sequences were localized with the single strand-specific ribonucleases A, T1, T2 and S1. The data enabled the various minimal secondary structural models, proposed for the 3'-regions of the E. coli and S. cerevisiae RNAs, to be critically examined, and to demonstrate that the main common features of these models are correct. The results also reveal the presence and position of additional higher order structure in the renatured free RNA. It can be concluded that a high level of conservation of higher order structure has occurred during the evolution of the gram negative and gram positive eubacteria and the eukaryote in both the double helical regions and the "unstructured" regions. Several unusual structural features were detected. Multiple G X A pairings in two of the putative helices, which are compatible with phylogenetic sequence comparisons, are strongly supported by the occurrence of cobra venom ribonuclease cuts adjacent to, and in one case between, these pairings. Evidence is also provided for the stacking of an A X A pair within a double helix of the yeast RNA. Other special structural features include adenosines bulged out from double helices; such nucleotides, which are hyper-reactive, have been implicated in protein recognition in 5 S ribosomal RNA. The 3'-terminal regions of the RNAs are particularly important for the functioning of the ribosome. They are involved in mRNA, tRNA and ribosomal factor binding. The results reveal that while the functionally important RNA sequences tend to be conserved, they are not always accessible in the free RNA; the pyrimidine-rich "Shine and Dalgarno" sequence, for example, which is involved in mRNA recognition, occurs in a double helix in both eubacterial RNAs.


Induction of xylanases by sugar cane bagasse at different cell densities of Cellulomonas flavigena.
The effect of cell density on xylanolytic activity and productivity of Cellulomonas flavigena was evaluated under two different culturing conditions: fed-batch culture with discontinuous feed of sugar cane bagasse (SCB; condition 1) and glycerol fed-batch culture followed by addition of SBC as xylanases inducer (condition 2). The enzymatic profile of xylanases was similar in both systems, regardless of the initial cell density at time of induction. However, the xylanolytic activity changed with initial cell density at the time of induction (condition 2). The maximum volumetric xylanase activity increased with increased initial cell density from 4 to 34 g l(-1) but decreased above this value. The largest total volumetric xylanase productivity under condition 2 (1.3 IU ml(-1) h(-1)) was significantly greater compared to condition 1 (maximum 0.6 IU ml(-1) h(-1)). Consequently, induction of xylanase activity by SCB after growing of C. flavigena on glycerol at intermediate cell density can be a feasible alternative to improve activity and productivity of xylanolytic enzymes.


Successful penetration and bougie dilatation with Brockenbrough needle for severely calcified occlusion in superficial femoral artery.
A 75-year-old hemodialysis patient with right critical limb ischemia received endovascular therapy for a chronic total occlusion (CTO) in a diffusely calcified superficial femoral artery (SFA). During a retrograde approach, a Brockenbrough needle (BN) was able to penetrate the calcified hard plaque formed in the middle segment of the CTO. Moreover, bougie dilatation with the BN allowed balloon crossing and stent deployment, even after failure to pass a 2.0-mm monorail balloon across the plaque. These results suggest that the BN offers a new therapeutic option in the penetration and modification of severely calcified CTO in SFA.


Selective synthesis of 3-formylbenzofuran and 3-acylbenzofuran using a chalcone rearrangement strategy.
We developed a method for highly selective synthesis of two benzofuran isomers, by rearranging and subsequently transforming 2-hydroxychalcones. Depending on the reaction conditions, synthesis of 3-formylbenzofurans, unconventional products, and 3-acylbenzofurans was achieved through cyclized 2,3-dihydrobenzofurans obtained from the rearranged products. The facile synthesis of 3-formylbenzofurans facilitated synthesis of the natural product, puerariafuran, from the corresponding chalcone.


Dietary supplementation of garlic, propolis, and wakame improves recuperation in cadmium exposed Japanese medaka fish (Oryzias latipes).
Cadmium (Cd) toxic effects can overwhelm endogenous antioxidants and affect recuperation in fish. Thus, this study assessed antioxidative properties and ameliorating potentials of dietary supplemented garlic (D1), propolis (D2), and wakame (D3) on i) Cd bioconcentration, and ii) Cd induced toxicity during recuperation in Cd exposed Japanese medaka. In a 21-day exposure, control (0.0 mg Cd/L in water-C1) or Cd-treatment (0.3 mg Cd/L in water-T1) fish were fed medaka diets. Surviving fish in T1 were further depurated for 21-days and fed D1, D2, D3, or medaka diet (C2). Gill, liver, and muscle tissues were assessed weekly for Cd bioconcentration, metallothionein, (MT), superoxide dismutase (SOD), total glutathione (GSH), and lipid peroxidation (LPO). Results showed reduced antioxidant activity by significantly increasing tissue Cd and LPO, and significantly reducing SOD activity and GSH content in gill and muscle upon Cd exposure. In contrast, D1, D2, and D3 diets significantly reduced tissue Cd and LPO, while increasing contents of MT and GSH, and SOD activity, significantly. Other condition indices in D1, D2 and D3 groups were also significantly higher than those in C2 groups. In conclusion, dietary supplementation significantly increased recuperation and tissue functions in fish, in the order D1 > D2 > D3> C2.


Effects of brain monoamine depletion on thermoregulation, active avoidance, and food and water intake in rats.
Intraventricular administration of 6-OHDA or 5,6-DHT suppressed food intake, whereas their effect on active avoidance produced a suppression with the former and an enhancement with the latter. The increased water intake was specifically associated with 5,6-DHT treatment in rats.


Workplace wellbeing in an urban emergency department in Aotearoa New Zealand.
Staff wellbeing is important for individuals and considered integral to the development and maintenance of high performing health systems. Unless baseline parameters of wellbeing are known, the effects of any interventions to improve staff wellbeing will remain uncertain. By clarifying staff perceptions and objective measures of important factors related to wellbeing, our primary goal was to assess the baseline wellbeing of staff in our central city ED. A survey template, the WoWe@AED (Workplace Wellbeing at the Adult Emergency Department) was developed from several sources. Burnout was measured using the Copenhagen Burnout Inventory. Inclusion in the survey required that participants worked in our ED, all staff groups were eligible. The electronic survey was conducted between 22 January and 5 February 2018. Quantitative and qualitative data were analysed. Two hundred and seventy (71.1%) of 380 potential participants participated. All staff groups were represented. About 72.4% agreed/strongly agreed that Adult Emergency Department was an excellent place to work, 80.7% found their work meaningful. About 42.0% felt well informed regarding important decisions. One hundred and eighty-three (48.2%) out of 380 completed the Copenhagen Burnout Inventory. Overall, personal burnout was 42.1% (n = 77), work-related burnout was 35% (n = 64) and client-related burnout was 27.9% (n = 51). Females were significantly (<0.05) more likely to have burnout in some or all domains. Four key themes were identified: supportive teamwork and relationships; providing high quality patient care; resourcing; and workload issues. Baseline indices of wellbeing of the frontline staff in a central city ED have been documented. These have informed wellbeing interventions. Future measures can be compared against these data.


Randomized controlled trial to prevent postpartum depression in mothers on public assistance.
Postpartum depression (PPD) is a significant and common public health problem for women. To examine the efficacy of an intervention based on the principles of interpersonal therapy (IPT) in reducing the risk of PPD in pregnant women. Randomized controlled trial of 205 pregnant women who were 18 years old or older, on public assistance, and at risk for PPD. Participants (mean age=23; 38% Hispanic and 23% Black) were randomized to either the IPT group intervention (n=104) or the treatment as usual control (TAU) program (n=101). At 6 months, the overall depression rate in the intervention group (16%) was lower than the control group (31%) and the effect of the intervention was statistically significant at p<0.05. It is unknown if findings will generalize to a more heterogeneous sample of women than the current study, such as women from a range of socio-economic and cultural backgrounds, or marital status. There was a differential amount of contact between TAU and intervention conditions. An IPT based intervention during the prenatal period has the potential to reduce cases of PPD within 6 months postpartum in at risk mothers on public assistance.


Association Between Genetic Testing for Hereditary Breast Cancer and Contralateral Prophylactic Mastectomy Among Multiethnic Women Diagnosed With Early-Stage Breast Cancer.
Increasing usage of multigene panel testing has identified more patients with pathogenic or likely pathogenic (P or LP) variants in low-moderate penetrance genes or variants of uncertain significance (VUS). Our study evaluates the association between genetic test results and contralateral prophylactic mastectomy (CPM) among patients with breast cancer. We conducted a retrospective cohort study among women diagnosed with unilateral stage 0-III breast cancer between 2013 and 2020 who underwent genetic testing. We examined whether genetic test results were associated with CPM using multivariable logistic regression models. Among 707 racially or ethnically diverse women, most had benign or likely benign (B or LB) variants, whereas 12.5% had P or LP and 17.9% had VUS. Racial or ethnic minorities were twice as likely to receive VUS. Patients with P or LP variants had higher CPM rates than VUS or B or LB (64.8% v 25.8% v 25.9%), and highest among women with P or LP variants in high-penetrance genes (74.6%). On multivariable analysis, P or LP compared with B or LB variants were significantly associated with CPM (odds ratio = 4.24; 95% CI, 2.48 to 7.26). Women with P or LP variants on genetic testing were over four times more likely to undergo CPM than B or LB. Those with VUS had similar CPM rates as B or LB. Our findings suggest appropriate genetic counseling and communication of cancer risk to multiethnic breast cancer survivors.


Corrigendum to: Hepatitis B Virus Genotyping Among Patients With Cirrhosis [ Published in Jundishapur J Microbiol. 2015 March; 8(3): e14571].
[This corrects the article DOI: 10.5812/jjm.14571.].


In situ construction of Fe3Al@Al2O3 core-shell particles with excellent electromagnetic absorption.
To obtain Fe3Al@Al2O3 core-shell absorbents, DO3-type Fe3Al powder was thermal treated in an argon atmosphere containing a trace amount of oxygen at different temperatures. Since Al atoms have a higher diffusion rate than that of the Fe atoms, Al atoms can migrate to the surface of the Fe3Al particle and in-situ convert to Al2O3 nanoparticles during the thermal treatment process. With the increase of the thermal treatment temperature, the Al2O3 nanoparticles grow larger, exhibiting different microwave absorption properties. In particular, the Fe3Al@Al2O3 obtained by controllable oxidation at 800 ℃ exhibits the best microwave absorption properties, with the minimum reflection loss of -34 dB at 11.5 GHz when the thickness is 2 mm, and the bandwidth below -10 dB is as broad as 6.7 GHz. Since a dielectric Al2O3 shell with a proper thickness can increase the impedance matching ratio of the Fe3Al absorbent, more electromagnetic waves can come into the absorbent. In addition, the magnetic Fe3Al core can efficiently attenuate the absorbed electromagnetic waves by dimensional resonance and natural resonance.


Cyclosporin A pretreatment in a rat model of warm ischaemia/reperfusion injury.
These studies investigated the role of apoptosis following ischaemia/reperfusion (I/R) injury to the liver and the effect of pretreatment with Cyclosporin A. Male Sprague-Dawley rats received 30 min of warm ischaemia followed by a period of reperfusion of 6 h. Rats were given olive oil or Cyclosporin A (30 mg/kg p.o.) the day before surgery. Neutrophil numbers were assessed in haematoxylin-eosin-stained sections of liver. In situ staining of sections using TdT-mediated dUTP-fluorescein nick-end labelling was carried out to determine the extent of apoptosis, followed by electron microscopy. Semi-quantitative polymerase chain reaction (PCR) analysis of the transcript for Fas antigen was performed. High levels of apoptosis were observed in I/R injury, which were greatly ameliorated in Cyclosporin A-pretreated groups. PCR analysis indicated a reduction in the level of expression of Fas transcript in Cyclosporin A-treated rats. Histological analysis showed a significant increase in the number of neutrophils infiltrating I/R-injured tissue (62 +/- 10.69, n=16), which was markedly reduced by Cyclosporin A pretreatment (16 +/- 7, n=6, P<0.05). These results indicate a role of parenchymal apoptosis in the pathogenesis of I/R injury, which occurs in association with neutrophil infiltration, both of which can be significantly reduced by Cyclosporin A pretreatment.


A novel triple dartous fixation in hypospadias repair.
This study investigates the impact of the use of dartos covering to augment the neourethra on functional and cosmetic results. To evaluate a novel technique demonstrating how to fix dartos flap to cover the neourethra as a barrier in hypospadias repair. This study comprised 204 male patients with different degree of hypospadias (DPH = 132, coronal hypospadias = 46, MPH = 26). Their ages ranged from 1 to 23 ys (mean age = 2 ys). Penile chordee was in (DPH = 45, coronal hypospadias = 33, MPH = 26). All patients had abnormal downward directed urinary stream. Hypospadias repair was performed by the classic TIP technique in addition to our novel modification of dartos covering. Patients were submitted to 5 years of follow-up including clinical examination of the functional and cosmetic parameters. Success rates were reported in 200 patients, 3 patients had complications with subcoronal urethrocutaneous fistula and one patient with complete repair disruption. Triple dartos fixation is a simple technique for all hypospadiologists to minimize urethrocutaneous fistula as common complication of hypospadias repair with good functional and cosmetic results.


Integrating cultivation and metagenomics for a multi-kingdom view of skin microbiome diversity and functions.
Human skin functions as a physical barrier to foreign pathogen invasion and houses numerous commensals. Shifts in the human skin microbiome have been associated with conditions ranging from acne to atopic dermatitis. Previous metagenomic investigations into the role of the skin microbiome in health or disease have found that much of the sequenced data do not match reference genomes, making it difficult to interpret metagenomic datasets. We combined bacterial cultivation and metagenomic sequencing to assemble the Skin Microbial Genome Collection (SMGC), which comprises 622 prokaryotic species derived from 7,535 metagenome-assembled genomes and 251 isolate genomes. The metagenomic datasets that we generated were combined with publicly available skin metagenomic datasets to identify members and functions of the human skin microbiome. The SMGC collection includes 174 newly identified bacterial species and 12 newly identified bacterial genera, including the abundant genus 'Candidatus Pellibacterium', which has been newly associated with the skin. The SMGC increases the characterized set of known skin bacteria by 26%. We validated the SMGC metagenome-assembled genomes by comparing them with sequenced isolates obtained from the same samples. We also recovered 12 eukaryotic species and assembled thousands of viral sequences, including newly identified clades of jumbo phages. The SMGC enables classification of a median of 85% of skin metagenomic sequences and provides a comprehensive view of skin microbiome diversity, derived primarily from samples obtained in North America.


A Randomized Trial to Assess Regional Left Ventricular Function After Stent Implantation in Chronic Total Occlusion: The REVASC Trial.
The aim of this study was to investigate whether percutaneous coronary intervention (PCI) of chronic total occlusions (CTOs) improves left ventricular function. The benefit of PCI in CTOs is still controversial. Patients with CTOs who were candidates for PCI were eligible for the study and were randomized to PCI or no PCI of CTO. Relevant coexisting non-CTO lesions were treated as indicated. Patients underwent cardiac magnetic resonance imaging at baseline and at 6 months. The primary endpoint was the change in segmental wall thickening (SWT) in the CTO territory. Secondary endpoints were improvement of regional wall motion and changes in left ventricular volumes and ejection fraction. Furthermore, major adverse coronary events after 12 months were assessed. The CTO PCI group comprised 101 patients and the no CTO PCI group 104 patients. The change in SWT did not differ between the CTO PCI (4.1% [interquartile range: 14.6 to 19.3]) and no CTO PCI (6.0% [interquartile range: 8.6 to 6.0]) groups (p = 0.57). Similar results were obtained for other indexes of regional and global left ventricular function. Subgroup analysis revealed that only in patients without major non-CTO lesions (basal SYNTAX [Synergy Between Percutaneous Coronary Intervention With Taxus and Cardiac Surgery] score ≤13) CTO PCI was associated with larger improvement in SWT than no CTO PCI (p for interaction = 0.002). Driven by repeat intervention, major adverse coronary event rates at 12 months were significantly lower in the CTO PCI group (16.3% vs. 5.9%; p = 0.02). No benefit was seen for CTO PCI in terms of the primary endpoint, SWT, or other indexes of left ventricular function. CTO PCI resulted in clinical benefit over no CTO PCI, as evidenced by reduced major adverse coronary event rates at 12 months.


N-acetylcysteine ameliorates carbofuran-induced alterations in lipid composition and activity of membrane bound enzymes.
The present work investigates the protective effects of N-acetylcysteine (NAC) on carbofuran-induced alterations in lipid composition and activity of membrane bound enzymes (Na+-K+-ATPase and Ca2+-ATPase) in the rat brain. Animals were exposed to carbofuran at a dose of 1 mg/kg body weight, orally, for a period of 28 days. A significant increase in lipid peroxidation in terms of TBARS was observed in brain after carbofuran exposure. NAC administration (200 mg/kg body weight) on the other hand lowered the carbofuran-induced lipid peroxidation to near normal. The increased lipid peroxidation following carbofuran exposure was accompanied by a significant decrease in the levels of total lipids, which is attributed to the reduction in phospholipid levels. Furthermore, NAC administration had a beneficial effect on carbofuran-induced alterations in lipid composition. The ratio of cholesterol to phospholipid, a major determinant of membrane fluidity, was increased in response to carbofuran exposure. This was associated with decreased activity of Na+-K+-ATPase and Ca2+-ATPase. NAC was observed to offer protection by restoring the cholesterol to phospholipid ratio along with the activity of Na+-K+-ATPase and Ca2+-ATPase. The results clearly suggest that carbofuran exerts its neurotoxic effects by increasing lipid peroxidation, altering lipid composition and activity of membrane bound enzymes. NAC administration ameliorated the effects of carbofuran suggesting its potential therapeutic effects in carbofuran neurotoxicity.


Risk and predictors of mortality after implantable cardioverter-defibrillator implantation in patients with sarcoid cardiomyopathy.
Implantable cardioverter-defibrillators (ICDs) are recommended for patients with cardiac sarcoidosis (CS) with an indication for pacing, prior ventricular arrhythmias, cardiac arrest, or left ventricular ejection fraction <35%, but data on outcomes are limited. Using data from the National Cardiovascular Data Registry ICD Registry between April 1, 2010 and December 31, 2015, we evaluated a propensity matched cohort of CS patients implanted with ICDs versus non-ischemic cardiomyopathies (NICM). We compared mortality using Kaplan-Meier survival curves and Cox proportional hazards models. We identified 1,638 patients with CS and 8,190 propensity matched patients with NICM. The rate of death at 1 and 2 years was similar in patients with CS and patients with NICM (5.2% vs 5.4%, P = 0.75 and 9.0% vs 9.3%, P = 0.72, respectively). After adjusting for other covariates, patients with CS had similar mortality at 2 years after ICD implantations compared with NICM patients (RR 1.03, 95% CI 0.87-1.23). Among patients with CS, multivariable logistic regression identified 6 factors significantly associated with increased 2-year mortality: presence of heart failure (HR 1.92, 95% CI 1.44-3.22), New York Heart Association (NYHA) Class III heart failure (HR 1.68, 95% CI 1.16-2.45), NYHA Class IV heart failure (HR 3.08, 95% CI 1.49-6.39), atrial fibrillation/flutter (HR 1.66, 95% CI 1.17-2.35), chronic lung disease (HR 1.64, 95% CI 1.17-2.29), creatinine >2.0 mg/dL (HR 4.07, 95% CI 2.63-6.30), and paced rhythm (HR 2.66, 95% CI 1.07-6.59). Mortality following ICD implantation was similar in CS patients compared with propensity matched NICM patients. Presence of heart failure, NYHA class, atrial fibrillation/flutter, chronic lung disease, renal dysfunction, and paced rhythm at time of implantation were all predictors of increased 2-year mortality among CS patients with ICDs.


Endourology treatment of giant ureteral stone: Distal ureteral stone lithotripsy with nephroscope and shockpulse.
There has been no article that publish the utilization of nephroscope to lithotripsy in removing giant distal ureteral stone. Here we reported a case of large multiple distal ureteral stones treated using nephroscope and litotripsy with shockpulse in a 53-year-old woman. The biggest stone measuring 40 × 35 mm and 5 × 5 mm for the smallest one. After initial cystoscopy and ureteric dilatation, nephroscope 24fr was inserted into the distal ureter. DJ stent was applied. No residual stone and complication were found. After four weeks the stent was removed and the ureteral lumen was normal in size.


Rabphilin localizes with the cell actin cytoskeleton and stimulates association of granules with F-actin cross-linked by {alpha}-actinin.
In endocrine cell, granules accumulate within an F-actin-rich region below the plasma membrane. The mechanisms involved in this process are largely unknown. Rabphilin is a cytosolic protein that is expressed in neurons and neuroendocrine cells and binds with high affinity to members of the Rab3 family of GTPases localized to synaptic vesicles and dense core granules. Rabphilin also interacts with alpha-actinin, a protein that cross-links F-actin into bundles and networks and associates with the granule membrane. Here we asked whether rabphilin, in addition to its granule localization, also interacts with the cell actin cytoskeleton. Immunofluorescence and immunoelectron microscopy show that rabphilin localizes to the sub-plasmalemmal actin cytoskeleton both in neuroendocrine and unspecialized cells. By using purified components, it is found that association of rabphilin with F-actin is dependent on added alpha-actinin. In an in vitro assay, granules, unlike endosomes or mitochondria, associate with F-actin cross-linked by alpha-actinin. Rabphilin is shown to stimulate this process. Rabphilin enhances by approximately 8-fold the granule ability to localize within regions of elevated concentration of cross-linked F-actin. These results suggest that rabphilin, by interacting with alpha-actinin, organizes the cell cytoskeleton to facilitate granule localization within F-actin-rich regions.


Initiating ribosomes and a 5'/3'-UTR interaction control ribonuclease action to tightly couple B. subtilis hbs mRNA stability with translation.
We previously showed that ribosomes initiating translation of the B. subtilis hbs mRNA at a strong Shine-Dalgarno sequence block the 5' exoribonuclease RNase J1 from degrading into the coding sequence. Here, we identify new and previously unsuspected features of this mRNA. First, we identify RNase Y as the endoribonuclease that cleaves the highly structured 5'-UTR to give access to RNase J1. Cleavage by RNase Y at this site is modulated by a 14-bp long-range interaction between the 5'- and 3-UTRs that partially overlaps the cleavage site. In addition to this maturation/degradation pathway, we discovered a new and ultimately more important RNase Y cleavage site in the very early coding sequence, masked by the initiating ribosome. Thus, two independent pathways compete with ribosomes to tightly link hbs mRNA stability to translation initiation; in one case the initiating ribosome competes directly with RNase J1 and in the other with RNase Y. This is in contrast to prevailing models in Escherichia coli where ribosome traffic over the ORF is the main source of protection from RNases. Indeed, a second RNase Y cleavage site later in the hbs ORF plays no role in its turnover, confirming that for this mRNA at least, initiation is key.


Acceptance of marginal liver donors increases the volume of liver transplant: early results of a single-center experience.
To expand the donor pool, clinicians are continually modifying criteria to accept organs, particularly those in the so-called expanded or marginal donor pool. The concept and definition of a marginal donors continues to evolve. The impact of their use is the result of a combination of donor and recipient factors. Most clinicians accept steatosis above 30%, donor age over 60 years, prolonged ischemia time, prolonged intensive care unit stay, hypernatremia, previous cardiac arrest, prolonged episodes of hypotension, large use of inotrope drugs, and elevated liver function tests as criteria for designation of a marginal organ. In June 2003, we started to use marginal donors each year tripling the number of transplants per year at our center.


Border between aplastic anemia and myelodysplastic syndrome.
Distinguishing between acquired aplastic anemia (AA) and myelodysplastic syndrome (MDS) with a low blast cell percentage is often difficult and problematic, as both diseases are syndromes primarily defined by morphological findings, and their diagnostic criteria do not necessarily reflect the pathophysiology of their bone marrow (BM) failure. As a result, many patients with benign BM failure that should be managed as AA are diagnosed as having MDS, due to the absence of BM hypocellularity and the presence of dysplastic signs in the BM, and are treated inappropriately with toxic therapies, such as hypomethylating agents, and stem cell transplantation from unrelated donors. BM failure syndromes need to be managed in ways appropriate to their pathophysiology, which is more accurately determined by using markers such as the presence of glycosylphosphatidylinositol-anchored protein-deficient cells and HLA-A lacking leukocytes. We recently found that plasma thromobopoietin level is one of the most useful markers for distinguishing benign and pre-leukemic BM failure syndromes.


Comparison of evapotranspiration components and water-use efficiency among different land use patterns of temperate steppe in the Northern China pastoral-farming ecotone.
Water-use efficiency (WUE), which links carbon and water cycles, is an important indicator of assessing the interactions between ecosystems and regional climate. Using chamber methods with and without plant removal treatments, we investigated WUE and evapotranspiration (ET) components in three ecosystems with different land-use types in Northern China pastoral-farming ecotone. In comparison, ET of the ecosystems with grazing exclusion and cultivating was 6.7 and 13.4 % higher than that of the ecosystem with free grazing. The difference in ET was primarily due to the different magnitudes of soil water evaporation (E) rather than canopy transpiration (T). Canopy WUE (WUEc, i.e., the ratio of gross primary productivity to T) at the grazing excluded and cultivated sites was 17 and 36 % higher than that at the grazing site. Ecosystem WUE (WUEnep, i.e., the ratio of net ecosystem productivity to ET) at the cultivated site was 34 and 28 % lower in comparison with grazed and grazing excluded stepped, respectively. The varied leaf area index (LAI) of different land uses was correlated with microclimate and ecosystem vapor/carbon exchange. The LAI changing with land uses should be the primary regulation of grassland WUE. These findings facilitate the mechanistic understanding of carbon-water relationships at canopy and ecosystem levels and projection of the effects of land-use change on regional climate and productivity.


Half-encapsulated Au nanoparticles by nano iron oxide: promoted performance of the aerobic oxidation of 1-phenylethanol.
Au nanoparticles half-encapsulated in nano iron oxide are prepared and loaded on alumina as a support. The donation of electrons from nano iron oxide to Au nanoparticles is detected and both the properties of gold and iron oxide are adjusted by the donation. The properties are different from the bulk iron oxide supported gold catalysts, in which the iron oxide is little influenced by the electronic interaction between the two components. The catalyst shows noticeably promoted activity for the aerobic oxidation of 1-phenylethanol over Au-Al2O3 and Au-bulk FeOx. The enhanced catalytic behavior may result from the cooperative effect between the Au nanoparticles and nano iron oxide.


Population data on 10 non-CODIS STR loci in Japanese population using a newly developed multiplex PCR system.
This paper describes a newly devised autosomal short tandem repeat (STR) multiplex polymerase chain reaction (PCR) systems for 10 loci (D1S1656, D2S1353, D8S1132, D12S1090, D14S608, D18S535, D19S253, D20S480, D21S226, and D22S689) unlinked to the core STR loci (non-CODIS loci). Of 252 samples taken from the Japanese population, PCR products ranged in length from 107 bp to 319 bp. No significant deviations from Hardy-Weinberg equilibrium were observed at any of the 10 loci. The accumulated power of discrimination and power of exclusion for the 10 loci were 0.999999999998 and 0.99991, respectively. We conclude that the present multiplex system for the 10 non-CODIS loci represents a powerful tool for forensic applications.


Cadmium removal from human plasma by Cibacron Blue F3GA and thionein incorporated into polymeric microspheres.
Poly(2-hydroxyethylmethacrylate-ethyleneglycoldimethacrylate) [poly(HEMA-EGDMA)] microspheres carrying Cibacron Blue F3GA and/or thionein were prepared and used for the removal of cadmium ions Cd(II) from human plasma. The poly(HEMA-EGDMA) microspheres, in the size range of 150-200 microm in diameter, were produced by a modified suspension copolymerization of HEMA and EGDMA. The reactive triazinyl dye-ligand Cibacron Blue F3GA was then covalently incorporated into the microspheres. The maximum dye incorporation was 16.5 micromol/g. Then, thionein was bound onto the Cibacron Blue F3GA-incorporated microspheres under different conditions. The maximum amount of thionein bound was 14.3 mg/g. The maximum amounts of Cd(II) ions removed from human plasma by poly(HEMA-EGDMA)-Cibacron Blue F3GA and poly(HEMA-EGDMA)-Cibacron Blue F3GA-thionein were of 17.5 mg/g and 38.0 mg/g, respectively. Cd(II) ions could be repeatedly adsorbed and desorbed with both types of microspheres without significant loss in their adsorption capacity.


Cost-benefit analysis of Haemophilus influenzae type b prevention: conjugate vaccination at eighteen months of age.
A cost-benefit analysis of Haemophilus influenzae type b disease preventive strategies was updated to consider evaluation of the H. influenzae type b polysaccharide-diphtheria toxoid conjugate vaccine (PRP-D) and H. influenzae type b oligosaccharide-mutant diphtheria toxin conjugate vaccine (HbOC) for children at 18 months of age. The analysis was done from the perspective of society as a whole. The economic costs of H. influenzae type b disease in the 1988 United States birth cohort would be $2.546 billion (1988 U.S. dollars) in the absence of preventive efforts. If 60% of all children could be vaccinated with PRP-D or HbOC at 18 months of age, this strategy would save $207.1 million ($88.22 savings/vaccinee; $43,605 cost/case prevented; 3.57/1 benefit-to-cost ratio) under base case model assumptions. Universal PRP-D or HbOC vaccination at 18 months of age would prevent 1845 cases of invasive H. influenzae type b disease. The break-even efficacy for universal PRP-D or HbOC vaccination at 18 months of age was 22.7%. We conclude that, under the model base case assumptions, universal PRP-D or HbOC vaccination at 18 months of age is sufficiently efficacious so that the costs of vaccination would be more than offset by decreased medical care costs for treating H. influenzae type b disease.


Respiratory hypersensitivity in guinea pigs sensitized to 1,6-hexamethylene diisocyanate (HDI): comparison of results obtained with the monomer and homopolymers of HDI.
This study used guinea pigs that were sensitized to the biuret or isocyanurate type homopolymers of 1,6 hexamethylene diisocyanate (HDI). Induction was either by intradermal injection or repeated inhalation exposures. For comparison, groups of guinea pigs were sensitized to monomeric HDI. Naive animals served as negative controls. Two and three weeks following induction, animals were challenged by inhalation with the hapten and homologous protein conjugate of the hapten, respectively. Assessments were based on changes in respiratory rate, serum IgG(1)-antibody titer, and influx of eosinophilic granulocytes in airways. Guinea pigs induced and challenged with the HDI-monomer did not display appreciable changes in respiratory rate, whilst the re-challenge with the HDI-protein conjugate caused unequivocal changes in respiratory patterns, including a marked bronchial influx of eosinophilic granulocytes. In contrast, animals induced and challenged with either the free or conjugated aerosols of HDI-homopolymers failed to elicit specific physiological or morphological pulmonary responses. IgG(1) antibodies were observed in all groups receiving monomeric HDI or HDI-homopolymers. Based on the comparative assessment of antibody titers following intradermal injections, it appeared that monomeric HDI was more potent to induce specific IgG(1) antibodies than the homopolymers of HDI. In summary, with respect to induction of allergy and asthma, the data presented here suggest that the homopolymeric forms of HDI appear to be less potent asthmagens, if any, than monomeric HDI.