Identification of a small-molecule ligand of β-arrestin1 as an inhibitor of stromal fibroblast cell migration accelerated by cancer cells.
Stromal fibroblasts, which occupy a major portion of the tumor microenvironment, play an important role in cancer metastasis. Thus, targeting of these fibroblasts activated by cancer cells (carcinoma-associated fibroblasts; CAFs) might aid in the improved treatment of cancer metastasis. NIH3T3 fibroblasts cocultured with MCF7 cells displayed enhanced migration compared to NIH3T3 fibroblasts cultured alone. We used this system to identify the small-molecule inhibitors responsible for their enhanced migration, a characteristic of CAFs. We selected β-arrestin1, which showed high expression in cocultured cells, as a molecular target for such inhibitors. Cofilin, a protein downstream of β-arrestin1, is activated/dephosphorylated in this condition. The small-molecule ligands of β-arrestin1 obtained by chemical array were then examined using a wound healing coculture assay. RKN5755 was identified as a selective inhibitor of activated fibroblasts. RKN5755 inhibited the enhanced migration of fibroblasts cocultured with cancer cells by binding to β-arrestin1 and interfering with β-arrestin1-mediated cofilin signaling pathways. Therefore, these results demonstrate the role of β-arrestin1 in the activation of fibroblasts and inhibiting this protein by small molecule inhibitor might be a potential therapeutic target for the stromal fibroblast activation (cancer-stroma interaction).


Induction of heat shock proteins in differentiated human and rodent neurons by celastrol.
Neurodegenerative disorders such as Alzheimer's disease, Parkinson's disease, and amyotrophic lateral sclerosis have been termed protein misfolding disorders that are characterized by the neuronal accumulation of protein aggregates. Manipulation of the cellular stress-response involving induction of heat shock proteins (Hsps) in differentiated neurons offers a therapeutic strategy to counter conformational changes in neuronal proteins that trigger pathogenic cascades resulting in neurodegenerative diseases. Hsps are protein repair agents that provide a line of defense against misfolded, aggregation-prone proteins. These proteins are not induced in differentiated neurons by conventional heat shock. We have found that celastrol, a quinine methide triterpene, induced expression of a wider set of Hsps, including Hsp70B', in differentiated human neurons grown in tissue culture compared to cultured rodent neuronal cells. Hence the beneficial effect of celastrol against human neurodegenerative diseases may exceed its potential in rodent models of these diseases.


Serum Uric Acid Level among Acute Stroke Patients.
Stroke is one of the leading causes of death and disability in developed as well as developing countries like Bangladesh. Elevated serum uric acid levels may predict an increased risk for cerebro-vascular (CV) events including stroke. Aim of the study was to measure the serum uric acid level among stroke patients and determine the relationship between serum uric acid level and stroke. This descriptive, cross-sectional study was carried out in Department of Medicine, Mymensingh Medical College Hospital, Mymensingh, Bangladesh to measure serum uric acid level among 102 stroke patients in a period of one year by using non-probability sampling procedure. Finally, collected data were analyzed using SPSS software Version 17.0. It was observed that the mean age of patients was 60.87±8.05 years, of them 80(78.43%) patients were male and the rest 22(21.57%) were female. About 66(64.70%) of respondents were in age group 60 years and above, while 36(35.30%) were in age group 59 years and below. At least 23(22.55%) of stroke patients had elevated serum uric acid with a mean serum uric acid level of 5.18mg/dl and standard deviation 1.26mg/dl. About 23(27.38%) patients in ischemic stroke had elevated serum uric acid whereas 18(100%) patients in hemorrhagic stroke had normal uric acid level. Uric acid level was elevated in ischemic stroke than haemorrhagic stroke patients (p<0.001). High uric acid level may be considered as a risk factor in patients with acute ischemic stroke.


The fertility impact of spousal separation.
The impact of spousal separation on marital fertility in a natural fertility context is investigated. REPMOD, a procedure for computer simulation of reproductive processes, is employed. A given length of separation affects fertility more, the less it overlaps with time during which the woman is not at risk of conception. Thus the "efficiency" of separation as a constraint on fertility increases as its duration increases, the duration of postpartum amenorrhea decreases, and fecundability decreases. Effects of spousal separation in actual situations can be estimated using results of these simulations as inputs.


Development towards Compact Nitrocellulose-Based Interferometric Biochips for Dry Eye MMP9 Label-Free In-Situ Diagnosis.
A novel compact optical biochip based on a thin layer-sensing surface of nitrocellulose is used for in-situ label-free detection of metalloproteinase (MMP9) related to dry eye disease. In this article, a new integrated chip with different interferometric transducers layout with an optimal sensing surface is reported for the first time. We demonstrate that specific antibodies can be immobilized onto these transducers with a very low volume of sample and with good orientation. Many sensing transducers constitute the presented biochip in order to yield statistical data and stability in the acquired measurements. As a result, we report the recognition curve for pure recombinant MMP9, tests of model tears with MMP9, and real tear performance from patients, with a promising limit of detection.


Identification of Four Novel COL4A5 Variants and Detection of Splicing Abnormalities in Three Chinese X-Linked Alport Syndrome Families.
Chronic renal disease associated with X-linked Alport syndrome (XLAS) is relatively rare. However, due to the lack of specificity in the pathologic and clinical manifestations of the disease, it is easy to be misdiagnosed. In this study, we included three Chinese families with XLAS and used targeted NGS to find gene variants. In family X1, the 36-year-old male proband had hematuria, massive proteinuria, sensorineural deafness and ESRD at 33. In silico prediction showed the novel c.1424-4C > G variant reduced the score of the normal 3' splice site from 0.47 to 0.00 (according to BDGP). Transcriptional analysis from his peripheral blood cells indicated that it caused the insertion of an amino acid [p.(Lys474_Gly475insVal)]. In family X2, the proband was a 32-year-old male, who had hematuria, proteinuria, hypertension, hearing loss and progressed into ESRD at 30 years. He carried a novel missense variant c.2777G > T p.(Gly926Val). In family X3, the proband, a 16-year-old male, had hematuria, massive proteinuria, sensorineural deafness and ESRD; the results of renal pathological findings were consistent with AS. He carried a novel variant c.4529-2A > T, so did his mother with ESRD and probable XLAS. Bioinformatic analysis with BDGP showed that it abolished the acceptor site from 0.83 to 0.00. RT-PCR analysis from his kidney tissue indicated that it caused exon 50 skipping and exon 50 skipping along with inserting a cryptic exon derived from intron 49 p.[Gly1510Aspfs*11, Gly1510Alafs*35]. Another novel missense variant c.1552G > A p.(Gly518Arg) was identified in his mother and his aunt. No skewed X-chromosome inactivation was involved in these two female patients. In conclusion, four novel variants in COL4A5 were identified and transcriptional analysis is essential to investigate the pathogenicity of intronic variants. Thus we found a rare event in a female patient with XLAS caused by two COL4A5 variants in trans.


Isolation and characterization of verocytotoxin-producing Escherichia coli O157 strains from Dutch cattle and sheep.
In the periods from July to November 1995 and 1996, fecal samples from Dutch cattle and sheep were collected at the main slaughterhouses of The Netherlands, located at different geographic sites. The samples were examined for the presence of verocytotoxin (VT)-producing Escherichia coli (VTEC) of serogroup 0157. E. coli O157 strains could be isolated from 57 (10.6%) of 540 adult cattle, 2 (0.5%) of 397 veal calves, 2 (3.8%) of 52 ewes, and 2 (4.1%) of 49 lambs. Immunomagnetic separation with O157-specific-antibody-coated beads appeared to be significantly more sensitive than conventional plating for detection of the organism in feces. With the exception of two isolates from adult cattle which appeared to be negative for VT genes, all animal isolates were positive for both VT (VT1 and/or VT2) and E. coli attaching-and-effacing gene sequences, and therefore, they were regarded as potential human pathogens. Although genomic typing by pulsed-field gel electrophoresis revealed a wide variety of distinct restriction patterns, comparison of the 63 animal isolates with 33 fecal O157 VTEC strains previously isolated from humans with the diarrhea-associated form of the hemolytic-uremic syndrome by their phage types and VT genotypes showed a marked similarity between animal and human isolates: 30 (90.9%) of the 33 human isolates appeared to be of E. coli O157 strain types also isolated from cattle and sheep. It was concluded that Dutch cattle and sheep are an important reservoir of E. coli O157 strains that are potentially pathogenic for humans.


Castleman's disease: cytomorphologic and cytochemical features of a case.
The cytomorphologic features of a case of giant lymph node hyperplasia, hyaline-vascular type, obtained by fine needle aspiration is described. Cytochemical studies of the vascular structures present in the aspirate suggest their origin as capillaries.


A novel role of TGFBI in macrophage polarization and macrophage-induced pancreatic cancer growth and therapeutic resistance.
Tumor-associated macrophages (TAMs), as a major and essential component of tumor microenvironment (TME), play a critical role in orchestrating pancreatic cancer (PaC) tumorigenesis from initiation to angiogenesis, growth, and systemic dissemination, as well as immunosuppression and resistance to chemotherapy and immunotherapy; however, the critical intrinsic factors responsible for TAMs reprograming and function remain to be identified. By performing single-cell RNA sequencing, transforming growth factor-beta-induced protein (TGFBI) was identified as TAM-producing factor in murine PaC tumors. TAMs express TGFBI in human PaC and TGFBI expression is positively related with human PaC growth. By inducing TGFBI loss-of-function in macrophage (MΦs) in vitro with siRNA and in vivo with Cre-Lox strategy in our developed TGFBI-floxed mice, we demonstrated disruption of TGFBI not only inhibited MΦ polarization to M2 phenotype and MΦ-mediated stimulation on PaC growth, but also significantly improved anti-tumor immunity, sensitizing PaC to chemotherapy in association with regulation of fibronectin 1, Cxcl10, and Ccl5. Our studies suggest that targeting TGFBI in MΦ can develop an effective therapeutic intervention for highly lethal PaC.


Efficient chemical potential evaluation with kinetic Monte Carlo method and non-uniform external potential: Lennard-Jones fluid, liquid, and solid.
The aim of this paper is to present a method of a direct evaluation of the chemical potential of fluid, liquid, and solid with kinetic Monte Carlo simulation. The method is illustrated with the 12-6 Lennard-Jones (LJ) system over a wide range of density and temperature. A distinctive feature of the methodology used in the present study is imposing an external potential on the elongated simulation box to split the system into two equilibrium phases, one of which is substantially diluted. This technique provides a reliable direct evaluation of the chemical potential of the whole non-uniform system (including that of the uniformly distributed dense phase in the central zone of the box), which, for example, is impossible in simulation of the uniform crystalline phase. The parameters of the vapor-liquid, liquid-solid, and fluid-solid transitions have been reliably determined. The chemical potential and the pressure are defined as thermodynamically consistent functions of density and temperature separately for the liquid and the solid (FCC) phases. It has been shown that in two-phase systems separated by a flat interface, the crystal melting always occurs at equilibrium conditions. It is also proved that in the limit of zero temperature, the specific heat capacity of an LJ crystal at constant volume is exactly 3Rg (where Rg is the gas constant) without resorting to harmonic oscillators.


Ordering kinetics in the active model B.
We undertake a detailed numerical study of the Active Model B proposed by Wittkowski et al., [Nature Commun. 5, 4351 (2014)]2041-172310.1038/ncomms5351. We find that the introduction of activity has a drastic effect on the ordering kinetics. First, the domain growth law shows a crossover from the usual Lifshitz-Slyozov growth law for phase separation (L∼t^{1/3}, where t is the time) to a novel growth law (L∼t^{1/4}) at late times. Second, the equal-time correlation function of the density field exhibits dynamical scaling for a given activity strength λ, but the scaling function depends on λ.


Solitary Myofibroma of the Mandible in a Six-Year Old-Child: Diagnosis of a Rare Lesion.
Myofibroma is a benign uncommon fibroblastic tumour originating from the soft tissue, bone and may affect both children and adults. Radiographically myofibroma presents as unilocular radiolucency. Histologically, typical biphasic cellular arrangement is noted. Immunohistochemical markers are useful for definitive diagnosis of this uncommon neoplasm. Here, a case of six-year-old male child with a localized swelling in the left body of mandible is presented. The clinical, radiological, histological and immunohistochemical findings were corroborative of Infantile Myofibroma.


The significance of laparoscopy in determining the optimal management plan for infertile patients with suspected tubal pathology revealed by hysterosalpingography.
The fallopian tube has numerous functions, including ovum pick-up, the place of fertilization of the ovum and cleavage of the embryo, and transfer of the embryo to the uterus. Tubal pathology impairs functions of the fallopian tube and reduces fertility. The degree of tubal pathology determines the possibility for fertility. The evaluation of the fallopian tube is necessary to determine the management plan of infertility. Hysterosalpingography (HSG) is often performed as a first line approach to assess tubal patency and the presence of adhesions; however, HSG has limitations in detecting tubal pathology. In the current study, we evaluated the significance of laparoscopy in determining the optimal management plan for infertile patients with suspected tubal pathology revealed by HSG. Between 1997 and 2009, 127 patients with suspected tubal pathology as demonstrated by HSG underwent laparoscopy at Kinki University Hospital, and a retrospective analysis was performed. Of 87 patients with unilateral tubal pathology revealed by HSG, 20 patients (23.0%) were given an indication for assisted reproductive technology (ART), based on the laparoscopic findings. Of 40 patients with bilateral tubal pathology revealed by HSG, 33 patients (82.5%) with bilateral tubal pathology detected by laparoscopy were given a high indication for ART. Laparoscopy enables exact evaluation of the fallopian tube and selection of the optimal management plan in infertile patients with suspected tubal pathology revealed by HSG. Therefore, laparoscopy should be performed in infertile patients with suspected tubal pathology revealed by HSG, as it is of diagnostic importance.


A systematic review of noninferiority margins in oncology clinical trials.
Aim: A systematic literature review was conducted to identify and characterize noninferiority margins for relevant end points in oncology clinical trials. Materials & methods: Randomized, controlled, noninferiority trials of patients with cancer were identified in PubMed and Embase. Results: Of 2284 publications identified, 285 oncology noninferiority clinical trials were analyzed. The median noninferiority margin was a hazard ratio of 1.29 (mean: 1.32; range: 1.05-2.05) for studies that reported time-to-event end points (n = 192). The median noninferiority margin was 13.0% (mean: 12.7%; range: 5.0-20.0%) for studies that reported response end points as absolute rate differences (n = 31). Conclusion: Although there was consistency in the noninferiority margins' scale, variability was evident in noninferiority margins across trials. Increased transparency may improve consistency in noninferiority margin application in oncology clinical trials.


[The value of resorting to 2 technics for detecting anti-mitochondrial antibodies].
The presence of antimitochondrial antibody was determined by an indirect immunofluorescence assay (IIFA) and a quantitative complement fixation test (CFT) in 2,302 sera. This survey enabled us to calculate the sensitivity, specificity, positive and negative predictive values of the two techniques. It seemed valuable to use both techniques as the IIFA was more sensitive than the CFT whilst the CFT was more specific than the IIFA.


[Extraperitoneal prosthesis repair in the surgical treatment of inguinal hernias].
The authors analyse the current state of the art of the prosthetic repair of incisional hernia and the problems involved in positioning the prosthesis, comparing their own experience with the most recent literature. From January 1994 to June 2001, 50 patients were operated on for incisional hernia (28 males and 22 females); 12% had recurrent or re-recurrent incisional hernias. Defects smaller than 3 cm were repaired with a polypropylene plug; a double-layer polypropylene mesh placed in a preperitoneal position was used for defects measuring from 3 to 5 cm; in defects greater than 5 cm a double-layer mesh was placed behind the muscle layer. Fifteen patients were operated on under local anaesthesia. Only 22% required postoperative analgesia. The mean hospital stay was 3.95 days. Only 3 recurrences (6%) were recorded. On the basis of our experience it seems appropriate to repair incisional hernias when of small size, preferably under local anaesthesia, avoiding opening the hernia sac, using an extraperitoneal approach with an overlap technique that employs polypropylene.


[Radiation dosage in orthopedics -- a comparison of computer-assisted procedures].
Since the 1990s, computer-assisted methods have been well-established in the field of orthopedics and traumatology. In addition to the proven improvement in the field of surgical precisioncompared with conventional techniques, the decrease in the amount of emitted ionizing radiation inside of the operating room was cited to constitute an additional major advantage of computer-based navigation. The goal of this study was to quantify X-ray dose values exposed during both conventional methods and computer-assisted procedures, the latter being performed using a C-arm device SIREMOBIL Iso-C3D (Siemens Medical Solutions). The clinical measurements were supplemented by laboratory experiments using a pinpoint ionization chamber placed inside a standard lucite phantom. The clinical part of the study investigated the application of new, i.e. computer assisted, techniques in comparison with conventional procedures on in total 42 patients experiencing surgery either on their lower extremities (n = 14) or on the spine (n = 28). The techniques applied were: (1) conventional surgical procedure, (2) CT-based navigation, (3) C-arm navigation, and (4) Iso-C3D-navigation. The main focus of the clinical evaluation was the determination of radiation dose values by means of thermoluminescence dose meters, accompanied by the registration of the operation times of the X-ray tube during a surgical intervention. The conclusions from the clinical studies are in agreement with the results from the laboratory measurements. Computer-assisted techniques led to a significant reduction in X-ray dose exposure and drastically shortened irradiation times. The most pronounced decrease in the emission of ionizing radiation was achieved in case of the Iso-C3D-navigation, which has only recently been introduced into clinical practice, i.e. during runtime of this study.


The effects of toluene plus noise on hearing thresholds: an evaluation based on repeated measurements in the German printing industry.
The ototoxicity of occupational exposure to toluene plus noise was investigated in a longitudinal study in rotogravure printing and existing findings in the literature were evaluated. The study comprised four repeated examinations during 5 years and started with 333 male workers. Lifetime weighted average exposures (LWAE) to toluene and noise were determined from individual work histories and historic recordings; recent individual exposures were measured 10 times during the study (toluene, active sampling; noise, stationary measurements). Auditory thresholds were measured with pure tone audiometry at 0.125, 0.25, 0.5, 0.75, 1, 2, 3, 4, 6, 8, 12 kHz. Mean LWAE exposures to toluene and noise were 45+/-17 ppm plus 82+/-7 dB(A) for high toluene exposed and 10+/-7 ppm plus 82+/-4 dB(A) for low toluene exposed subjects, mean current exposures were 26+/-20 ppm plus 81+/-4 dB(A) and 3+/-3 ppm plus 82+/-4 dB(A). Mean exposure duration was 21.3+/-6.5 years for long exposed and 5.9+/-2.2 years for short exposed subjects. Repeated measurement analyses of variance did not reveal effects of toluene intensity, exposure duration and interactions between toluene intensity and noise intensity. Noise intensity [79+/-3 dB(A) vs. 84+/-1 dB(A)] was significant for auditory thresholds. A case concept utilising developments of individual auditory thresholds did not reveal significant toluene effects. Logistic models including age, exposure duration, toluene in ambient air, current noise and either hippuric acid or ortho-cresol (o-cresol) found only age to be significant for elevated OR of high frequency hearing loss. Due to missing toluene effects, it was concluded that the threshold level for developing hearing loss as a result of occupational exposure to toluene plus noise might be above the current limit of 50 ppm toluene.


Maternal stress or anxiety during pregnancy and the development of the baby.
We have recently carried out a study of 100 mothers at Queen Charlotte's Hospital, at 32 weeks' gestation, and shown that those who were most anxious had impaired blood flow through the uterine arteries. This may help to explain why the babies of very anxious mothers tend to be smaller or born earlier. We have also shown that there is a strong correlation between plasma levels of the stress hormone cortisol in the mother and in the fetus. If the pregnant mother has raised cortisol, this may have a direct effect on the development of the fetal brain, and affect the child's later responses to stress.


Dual natriuretic peptide system in experimental heart failure.
The objectives of this study were 1) to define in an experimental model of heart failure the time course of changes in plasma brain natriuretic peptide concentrations during the development of and recovery from heart failure, and 2) to relate the changes to changes in atrial natriuretic peptide concentration and hemodynamic status. Brain natriuretic peptide is a circulating peptide with homology to atrial natriuretic peptide. However, unlike the latter, its changes during heart failure and its relation to cardiac filling pressures have not been studied. Eight male mongrel dogs underwent right ventricular pacing at 250 beats/min for 3 weeks until heart failure occurred and were followed up during recovery for 4 weeks after cessation of pacing. Heart failure was characterized by an increase in both left ventricular and end-diastolic pressure (6.6 +/- 4.1 mm Hg at the control measurements to 35.1 +/- 5.9 mm Hg at 3 weeks, p < 0.01) and right atrial pressure (6.7 +/- 1.1 to 11.4 +/- 2.1 mm Hg, p < 0.01). Recovery was accompanied by a return of cardiac filling pressures to control level. The time course of change of arterial plasma brain natriuretic peptide concentration was similar to that of atrial natriuretic peptide. Plasma concentrations of both peptides increased after 1 week of pacing (16 +/- 4 pg/ml at the control measurement to 59 +/- 20 pg/ml at 1 week, p < 0.001 for brain natriuretic peptide and 84 +/- 55 to 856 +/- 295 pg/ml, p < 0.001 for atrial natriuretic peptide). The level of both peptides then stayed level with no further increase at 3 weeks and returned to the control value by 4 weeks of recovery. There was an excellent correlation between plasma concentrations of the two peptides (r = 0.86, p < 0.001) and between the two peptides and cardiac filling pressures. However, compared with atrial natriuretic peptide, plasma brain natriuretic peptide concentration had a smaller percent increase during evolving heart failure and a slower rate of decline at recovery. Brain and atrial natriuretic peptide constitute a dual natriuretic system and are both responsive to changes in cardiac filling pressures in heart failure. However, brain natriuretic peptide appears to be less responsive than atrial natriuretic peptide.


Oxidative stress markers in plasma and urine of prepubertal patients with type 1 diabetes mellitus.
Markers of oxidative stress were studied in plasma and urine of prepubertal patients with type 1 diabetes mellitus (DM1) with less than 5 years of disease (n = 27). The results were compared to healthy, age- and sex-matched control children (n = 27). Oxidative stress parameters evaluated included advanced oxidation protein products (AOPP), total peroxyl radical-trapping antioxidant parameter (TRAP), and F2-isoprostanes (8-epi-prostaglandin-F2: 8-isoPGF2alpha). No statistically significant differences were found for any of the oxidative stress markers assessed between patients with DM1 and controls. In addition, weight, height, and routine metabolic tests, including creatininemia and cholesterol levels, were similar between the groups. The lack of significant differences between healthy controls and patients with DM1 suggests that treatment is able to counteract the increase in free radical production.


No anesthetic or analgesic benefit of neostigmine 1 mg added to intravenous regional anesthesia with lidocaine 0.5% for hand surgery.
Neostigmine has shown analgesic benefit when used as an adjunct to epidural or intrathecal anesthesia and analgesia, but evidence of benefit in the peripheral nervous system is controversial. This study aimed to determine if neostigmine 1 mg added to intravenous regional anesthesia (IVRA) provided any advantage in terms of intraoperative anesthesia or postoperative analgesia. We recruited 54 patients booked for hand surgery into this randomized, double-blind study. For the IVRA technique, patients were administered 3 mg/kg of 0.5% lidocaine (maximum 45 mL). The treatment group (group N) had 1 mg neostigmine added to lidocaine before dilution. The control group (group C) had no additives to the IVRA solution. At the completion of surgery and after transfer to the recovery room, patients had verbal response pain scores measured at 30 minutes, 1 hour, and 2 hours after cuff deflation. Time to first request for analgesic, side effects, and analgesic consumption at 24 hours were also recorded. Significantly more patients in group N had motor block at 5 and 10 minutes after injection of study solution. There were no other significant differences in sensory block onset, intraoperative anesthesia, postoperative analgesia, or adverse effects between groups. Neostigmine 1 mg provides no anesthetic or analgesic advantage when added to IVRA for upper limb surgery.


[Kaposi's sarcoma (an electron microscope case study)].
Endothelial histogenesis of sarcoma Kaposi was referred to in a case of 36-year old man studied by electron microscopy. Characteristic features corresponding to the literature were found: Weibel-Palade bodies, a ferritin-like material, phagocytosed erythrocytes, nuclear bodies, microfilaments and microtubules. Myelinated and unmyelinated axons were considered as remnants of original nervous tissue destroyed by the tumour or as nerve fibres accompanying proliferated vascular structures.


Adapted Kaizen: Multi-Organizational Complex Process Redesign for Adapting Clinical Guidelines for the Digital Age.
The need for a method to examine complex, multidisciplinary processes involving many diverse organizations initially led multiple US federal agencies to adopt the traditional Kaizen, a Lean process improvement method typically used within a single organization, to encompass multiple organizations each with its own leadership and priorities. First, the Centers for Medicare and Medicaid Services and the Office of the National Coordinator for Health Information Technology adapted Kaizen to federal agency processes for the development of electronic clinical quality measures. Later, the Centers for Disease Control and Prevention (CDC) further modified this adapted Kaizen during its Adapting Clinical Guidelines for the Digital Age (ACG) initiative, which aimed to improve the broader scope of guideline development and implementation. This is a methods article to document the adapted Kaizen method for future use in similar complex processes, illustrating how to apply the adapted Kaizen through CDC's ACG initiative and showing the reach achieved by using the adapted Kaizen method. The adapted Kaizen includes pre-Kaizen planning, a Kaizen event, and post-Kaizen implementation that accommodate multidisciplinary and multi-organizational participation. ACG included 5 workgroups that each developed products to support their respective scope: Guideline Creation, Informatics Framework, Translation and Implementation, Communication and Dissemination, and Evaluation. Despite challenges gathering diverse perspectives and balancing the competing priorities of multiple organizations, the ACG participants produced interrelated standards, processes, and tools-further described in separate publications-that programs and partners have leveraged. Use of a siloed approach may not have supported the development and dissemination of these products.


The RANK/RANKL/OPG triad in cancer-induced bone diseases.
The maintenance of skeletal integrity in a healthy individual requires a balanced regulation of the processes of bone formation, mediated by osteoblasts, and bone resorption, mediated by osteoclasts. This balanced process of bone remodeling becomes co-opted in the skeleton by tumor cells and this dramatically accelerates the process of remodeling and disrupts the normal equilibrium resulting in a spectrum of osteolytic to osteoblastic bone lesions. Certain tumor types, such as breast and prostate, frequently metastasize to the bone. It is now widely understood that the molecular triad--receptor activator of NF-kappaB ligand (RANKL), its receptor RANK, and the endogenous soluble RANKL inhibitor, osteoprotegerin (OPG)--play direct and essential roles in the formation, function, and survival of osteoclasts. Osteoclastic bone resorption contributes to the majority of skeletal sequelae, or skeletal-related events (SREs), in patients with bone metastases. In addition, osteoclastic bone resorption also contributes to the establishment of tumors in the skeleton. Therefore, blocking osteoclast activity and differentiation via RANKL inhibition may not only provide a beneficial treatment for skeletal complications of malignancy, but may also prevent bone metastases. In this review, we will first describe the operative role of osteoclasts and the RANK/RANKL/OPG triad in the pathophysiology of cancer-induced bone diseases, specifically solid tumor metastases to the bone. Secondly, we will describe a therapeutic approach that specifically targets the RANKL molecule.


Panhypopituitarism as an initial manifestation of primary central nervous system non-Hodgkin's lymphoma.
To report an unusual case of primary central nervous system non-Hodgkin's lymphoma in which the initial manifestation was panhypopituitarism. We present a retrospective case review and discuss similar cases from the literature. A 64-year-old woman with nausea, vomiting, diarrhea, and peripheral and periorbital edema was found to have panhypopituitarism. Magnetic resonance imaging showed minimal enlargement of the pituitary, and a transsphenoidal biopsy of the pituitary was nondiagnostic. Months later, abnormalities of extraocular movements developed. Repeated imaging and a second transsphe-noidal biopsy did not reveal the ultimate diagnosis. When further neurologic signs and symptoms subsequently developed, a right temporal open craniotomy was performed. It was not until this procedure, the patient's third biopsy, that the cause of her illness was discovered to be diffuse large cell lymphoma. Although idiopathic panhypopituitarism is a relatively common clinical entity, it remains a diagnosis of exclusion. The development of associated neurologic signs should prompt the clinician to initiate a new search for an underlying cause. This case underscores the protean manifestations of central nervous system lymphoma, both endocrine and neurologic, and the difficulties that may be encountered in attempts to establish a diagnosis.


Human Trafficking: When to Suspect in the Pediatric Emergency Department?
Human trafficking, which includes sexual exploitation and forced labor, affects youth throughout the world. Victims are subject to coercion, exploitation, and repeated trauma. Given the secretive nature of this abuse, it can be difficult to identify victims. Past experience shows that most victims interact with the healthcare system at some time while they are in the control of a trafficker, providing an opportunity for healthcare providers to identify, support, and intervene. In this CME review article, we describe human trafficking, its estimated prevalence, and the identification, evaluation, and management of patients who are victims of human trafficking. Finally, we provide additional resources for practitioners and patients.


Seroprevalence for 2117-like vesiviruses in Italian household dogs.
In 2003, a novel calicivirus, the vesivirus (VeV) strain 2117, was identified incidentally as a contaminant in Chinese Hamster Ovary (CHO) cell cultures by a German pharmaceutical company. Similar contaminations have been documented in three additional episodes, in bio-reactors used for production of recombinant drugs. More, recently 2117-like VeVs have also been identified at high prevalence in the stools from asymptomatic kennel dogs and only sporadically in diarrhoeic and healthy household dogs. In this study, antibodies for 2117-like viruses were detected in 21.5% of sera from household dogs, indicating that they are common viruses in the canine host.


Role of manganese in the pathogenesis of portal-systemic encephalopathy.
Amongst the potential neurotoxins implicated in the pathogenesis of hepatic encephalopathy, manganese emerges as a new candidate. In patients with chronic liver diseases, manganese accumulates in blood and brain leading to pallidal signal hyperintensity on T1-weighted Magnetic Resonance (MR) Imaging. Direct measurements in globus pallidus obtained at autopsy from cirrhotic patients who died in hepatic coma reveal 2 to 7-fold increases of manganese concentration. The intensity of pallidal MR images correlates with blood manganese and with the presence of extrapyramidal symptoms occurring in a majority of cirrhotic patients. Liver transplantation results in normalization of pallidal MR signals and disappearance of extrapyramidal symptoms whereas transjugular intrahepatic portosystemic shunting induces an increase in pallidal hyperintensity with a concomitant deterioration of neurological dysfunction. These findings suggest that the toxic effects of manganese contribute to extrapyramidal symptoms in patients with chronic liver disease. The mechanisms of manganese neurotoxicity are still speculative, but there is evidence to suggest that manganese deposition in the pallidum may lead to dopaminergic dysfunction. Future studies should be aimed at evaluating the effects of manganese chelation and/or of treatment of the dopaminergic deficit on neurological symptomatology in these patients.


Lipid-coated iron oxide nanoparticles for dual-modal imaging of hepatocellular carcinoma.
The development of noninvasive imaging techniques for the accurate diagnosis of progressive hepatocellular carcinoma (HCC) is of great clinical significance and has always been desired. Herein, a hepatocellular carcinoma cell-targeting fluorescent magnetic nanoparticle (NP) was obtained by conjugating near-infrared fluorescence to the surface of Fe3O4 (NIRF-Fe3O4) NPs, followed by coating the lipids consisting of tumoral hepatocytes-targeting polymer (Gal-P123). This magnetic NP (GPC@NIRF-Fe3O4) with superparamagnetic behavior showed high stability and safety in physiological conditions. In addition, GPC@NIRF-Fe3O4 achieved more specific uptake of human liver cancer cells than free Fe3O4 NPs. Importantly, with superpara-magnetic iron oxide and strong NIR absorbance, GPC@NIRF-Fe3O4 NPs demonstrate prominent tumor-contrasted imaging performance both on fluorescent and T2-weighted magnetic resonance (MR) imaging modalities in a living body. The relative MR signal enhancement of GPC@NIRF-Fe3O4 NPs achieved 5.4-fold improvement compared with NIR-Fe3O4 NPs. Therefore, GPC@ NIRF-Fe3O4 NPs may be potentially used as a candidate for dual-modal imaging of tumors with information covalidated and directly compared by combining fluorescence and MR imaging.


Soil anammox and denitrification processes connected with N cycling genes co-supporting or contrasting under different water conditions.
The anaerobic ammonium oxidizing bacteria (anammox) are the hidden culprit behind the excessive nitrogen loss under a favorable environment, since their detection and abundance get disturbed by several unknown factors. This study intends to find the gap between actual anammox working capacity under different water conditions and fertilizers in the laboratory. The 15N-isotopic tracer technique was used to measure anammox and denitrification rate, and anammox community structure was analyzed through high-throughput sequencing with cytochrome cd-1 nitrite reductase functional gene (an_nirS gene, initially found in Candidatus Scalindua). The experiment consisted of four treatments, i.e., (I) CK_ Control, (II) UR_Urea, (III) PM_Pig Manure, and (IV) SRF_ Slow release fertilizer, under two water conditions, i.e., (a) Continuous flooding_ CF, (b) Alternate wetting and drying_ AWD. Results showed that anammox under CF decreased over time by -40.24%, and denitrification increased up to 39.25%. However, anammox activity under AWD increased up to 10.62% with the availability of NO2-, and surprisingly accompanied by the reduction in denitrification loss (-31.97%), being the most critical factor. We found that soil nifH and AOB genes were strongly favorable for anammox activity, while we observed the presence of anammox and AOB genes co-existing at the same time in paddy soil. The high-throughput sequencing with an_nirS functional gene showed a much higher diversity of anammox genera ever reported, mostly uncultured and unidentified. We concluded that water management is more prominent than fertilizer for anammox, and the most critical factor is the duration of AWD cycle, because of short term air supply could boost anammox activity and gene abundance, and could reduce denitrifier activity as well as nirK gene abundance.


An examination of the psychometric properties of the attention deficit scales for adults with outpatient substance abusers.
A growing body of research has established a high co-morbidity in the rates of attention deficit/hyperactivity disorder (ADHD) and substance abuse. The co-occurrence of these two conditions and the negative impact of untreated ADHD on the course and treatment of substance abuse highlights the need for ADHD assessment in individuals seeking substance abuse treatment. In this article, we investigated the clinical utility of the Attention Deficit Scales for Adults (ADSA) in a sample of substance abusers in an outpatient treatment program by assessing its sensitivity, specificity, positive and negative predictive values, and likelihood ratios at two potential cut-off levels (161 and 181) using a DSM-IV based ADHD diagnosis as the criterion. The ADSA had strong reliability (Cronbach's alpha .93; Guttman split-half .92) and correlated well with DSM-IV diagnostic criteria. Sensitivity and specificity of the device were relatively strong at both cut-off levels, as were the positive and negative predictive values.


Regulation of experimental autoimmune encephalomyelitis by CD4+, CD25+ and CD8+ T cells: analysis using depleting antibodies.
Experimental Autoimmune Encephalomyelitis (EAE) can be induced in mice of the C57BL/6 strain by subcutaneous immunization with myelin/oligodendrocyte glycoprotein (MOG) peptide p35-55 in CFA, administered twice at an interval of one week and supplemented with Bordetella pertussis toxin given IV. Here, we studied the effect on the induction of EAE of depleting antibodies to CD4, CD8, or CD25 administered before either the first or the second dose of MOG p35-55. We found that anti-CD4 abolished EAE when given before the first immunization; anti-CD4 did not affect the disease when it was given before the second immunization. Anti-CD8 enhanced EAE induction when given before either of the two immunizations. Anti-CD25 enhanced EAE to the same degree as anti-CD8 when given before the first immunization, but anti-CD25 was even more effective in enhancing EAE when given before the second immunization. The anti-CD25 treatment led to significantly enhanced IFNgamma production by T cells responding to MOG p35-55 and persisting anti-MOG antibodies detectable 56 days after the first immunization. Administration of anti-CD8 or anti-CD25 abolished the need for pertussis toxin to induce EAE. These findings are compatible with the idea that CD4 T cells are required for the initial induction of EAE and that the disease is down-regulated by T cells expressing CD8 or CD25. These regulatory T cells exist prior to MOG immunization, but the CD25+ regulators appear to be further amplified by immunization.


Glucose-dependent glucose transporter 1 expression and its impact on viability of thyroid cancer cells.
Cancer cells exhibit an altered metabolism characterized by enhanced glycolysis and glucose consumption. In glucose‑addicted cancer cells upregulation of glucose transport across the plasma membrane is mediated by a family of facilitated glucose transporter proteins, particularly glucose transporter 1 (GLUT1). The aim of the present study was to investigate the impact of GLUT1 expression on glucose uptake and viability of FTC-133 and 8305c thyroid cancer cells growing in hypoglycemic, normoglycemic and hyperglycemic conditions. The results showed that the total expression of GLUT1 was higher in the two cell types growing in low glucose compared to cells growing in normoglycemia or hyperglycemia and this was correlated with AKT Ser473 phosphorylation but not with the expression of hypoxia inducible factor α (HIF1α). However, the membrane expression of GLUT1 was correlated with HIF1α expression. HIF1α expression was positively correlated with the glucose concentration in FTC-133 cells, whereas this expression was inversely correlated in 8305c cells. Glucose uptake was dependent on the membrane level of GLUT1 but not total GLUT1 expression. Downregulation of GLUT1 expression by RNAi in FTC-133 cells caused a reduction in glucose uptake but did not significantly affect cell viability. In the case of 8305c cells showing low endogenous GLUT1 expression and lack of HIF1α expression in normoxic conditions GLUT1 RNAi impacted cell viability. These data suggested that GLUT1 may be part of an AKT1-dependent mechanism allowing cells to survive in low levels of glucose. Glucose concentration inversely affected HIF1α expression and the level of GLUT1 in membrane as well as glucose uptake in FTC-133 and 8305c cells. The extent of GLUT1 impact on cell viability was also cell-type-dependent.


Successful development of squamous cell carcinoma and hyperplasia in RGEN-mediated p27 KO mice after the treatment of DMBA and TPA.
To evaluate the carcinogenicity of p27 knockout (KO) mice with RNA-guided endonuclease (RGENs)-mediated p27 mutant exon I gene (IΔ), alterations in the carcinogenic phenotypes including tumor spectrum, tumor suppressor proteins, apoptotic proteins and cell cycle regulators were observed in p27 (IΔ) KO mice after treatment with 7,12-Dimethylbenz[a]anthracene (DMBA) and 12-O-tetradecanoylphorbol-13-acetate (TPA)(DT) for 5 months. The target region (544~571 nt) in exon I of the p27 gene was successfully disrupted in p27 (IΔ) KO mice using the RGEN-induced non-homologous end joining (NHEJ) technique. After DT exposure for 5 months, a few solid tumors (identified as squamous cell carcinoma) developed on the surface of back skin of DT-treated p27 (IΔ) KO mice. Also, squamous cell hyperplasia with chronic inflammation was detected in the skin dermis of DT-treated p27 (IΔ) KO mice, while the Vehicle+p27 (IΔ) KO mice and WT mice maintained their normal histological skin structure. A significant increase was observed in the expression levels of tumor suppressor protein (p53), apoptotic proteins (Bax, Bcl-2 and Caspase-3) and cell-cycle regulator proteins (Cyclin D1, CDK2 and CDK4) in the skin of DT-treated p27 (IΔ) KO mice, although their enhancement ratio was varied. Taken together, the results of the present study suggest that squamous cell carcinoma and hyperplasia of skin tissue can be successfully developed in new p27 (IΔ) KO mice produced by RGEN-induced NHEJ technique following DT exposure for 5 months.


[Detoxification effects of two drugs in thallium -poisoned mice].
To observe the thallium eliminating effect of prussian blue, pentetate zinc trisodium (Zn-DTPA), and their combined use in the treatment of acute thallium poisoning in mice. Thallium poisoned mice were reproduced by oral administration of 0.2 ml thallous nitrate (3 mg/ml). They were assigned randomly to four groups according to the random number table method, namely, model group, prussian blue group, Zn-DTPA group and the combination therapy group, with 10 mice in each group. Prussian blue was administered orally [4.52 g×kg(-1)×d(-1), total four times], and Zn-DTPA was injected intraperitoneally [500 mg×kg(-1)×d(-1), one time]4 hours after giving thallium. The dosage of both drugs in combination treatment was as the same as described above. After treatment for 5 days, all the animals were sacrificed. Brain, intestine, kidney and liver of 1 mouse from each group were collected for pathological examination to observe the necrosis. Thallium contents of blood, brain, urine and feces from the other mice were determined. Pathological examination showed that the damage to intestine, kidney and liver was less obvious in treatment group compared with those of the model group. The effect was most obvious in the combination treatment group. However, brain damage was slightly improved. Thallium content in blood (mg/ml) of prussian blue group and the combination treatment group decreased obviously compared with the model group, and the decrease was more obvious in the combination treatment group (0.05 ± 0.01 vs. 0.18 ± 0.02). Thallium content in urine (mg/ml) and feces (mg/kg) was significantly increased after treatment, and the thallium elimination was most significant in the combined treatment group (urine: 11.34 ± 0.81 vs. 0.02 ± 0.01, feces: 13.11 ± 1.84 vs. 0.21 ± 0.07, both P < 0.01). Thallium content in brain was similar among all the groups. The single and combined use of prussian blue and Zn-DTPA could reduce the damage in intestine, kidney and liver. Combined use of prussian blue and Zn-DTPA for acute thallium poisoning mice is more efficacious than single use of either drug.


Evaluation of clinical safety and efficacy of left bundle branch area pacing in comparison with right ventricular septal pacing.
Left bundle branch area pacing (LBBaP) has recently emerged as a new physiological pacing strategy. The purpose of this study is to compare LBBaP with right ventricular sepal pacing (RVSP) in terms of their clinical safety and efficacy.From February 2019 to May 2020, consecutive pacing-indicated patients were prospectively enrolled and divided into 2 groups. Ventricular synchrony indexes such as QRS duration (QRSd), interventricular mechanical delay and septal-posterior wall motion delay, left ventricular function such as left ventricular end-diastolic diameter (LVEDD) and left ventricular ejection fraction (LVEF), pacing parameters, and complications were evaluated in the perioperative period and during follow-up.LBBaP was successful in 45 patients (88.2%), and finally 46 patients underwent RVSP. With LBBaP, ventricular electricalmechanical synchrony were similar to those of native-conduction system (P = .78). However, the ventricular electrical synchrony (QRSd, 108.47±7.64 vs 130.63±13.63ms, P < .001) and mechanical synchrony (interventricular mechanical delay, 27.68±4.33 vs 39.88±5.83, P < .001; septal-posterior wall motion delay, 40.39±23.21 vs 96.36±11.55, P < .001) in the LBBaP group were significantly better than those in the RVSP group. No significant differences in LVEDD (46 [44-48.5] vs 47 [44-52] mm, P = .49) and LVEF% (66 [62.5-70] vs 64 [61-68], P = .76) was observed between 2 groups at last follow-up. But, in the subgroup analysis, LVEDD was shorter (46 [44-49] vs 50 [47-58] mm, P = .03) and the LVEF% was higher (65 [62-68] vs 63 [58-65], P = .02) in the LBBaP-H (high ventricular pacing ratio >40%) group compared with RVSP-H group at last follow-up. There were lower capture thresholds (0.59±0.18V vs 0.71 ± 0.26 V, P = 0.01) at implantation in the LBBaP group than those in the RVSP group, with R-wave amplitudes and pacing impedances showing no significant difference between 2 groups. No serious complications were found in both 2 groups at implantation and follow-ups.This study confirms the clinical safety and efficacy of LBBaP, and it produces better ventricular electrical-mechanical synchrony than RVSP. The event of pacing-induced left ventricular dysfunction is lower in the LBBaP-H group than RVSP-H group.


Toll-like receptor-induced changes in glycolytic metabolism regulate dendritic cell activation.
Dendritic cells (DCs) are key regulators of innate and acquired immunity. The maturation of DCs is directed by signal transduction events downstream of toll-like receptors (TLRs) and other pattern recognition receptors. Here, we demonstrate that, in mouse DCs, TLR agonists stimulate a profound metabolic transition to aerobic glycolysis, similar to the Warburg metabolism displayed by cancer cells. This metabolic switch depends on the phosphatidyl inositol 3'-kinase/Akt pathway, is antagonized by the adenosine monophosphate (AMP)-activated protein kinase (AMPK), and is required for DC maturation. The metabolic switch induced by DC activation is antagonized by the antiinflammatory cytokine interleukin-10. Our data pinpoint TLR-mediated metabolic conversion as essential for DC maturation and function and reveal it as a potential target for intervention in the control of excessive inflammation and inappropriately regulated immune responses.


Evaluation of the patency rate of fibrous microvascular polyurethane prostheses after implantation in the rat aorta.
Two types of fibrous polyurethane prostheses with different compliances were implanted into the rat aorta with interrupted sutures and a running suture line. Excluding two technical failures, one unexplained death, and three infected prostheses, both groups (n = 21) showed a patency rate of 100%, up to 6 months after implantation. Rapid reendothelialisation occurred, and a stable neo-intima was formed. Compliance of the prostheses, as well as the suture technique used, has proven to be an unimportant factor in the rat aorta model, probably due to the low magnitude of the pulsations of the rat aorta. The experience of the surgeon with microsurgical techniques seems to be an underestimated factor in determining patency rates of microvascular prostheses.


Popliteal venous aneurysm.
Two new cases of popliteal venous aneurysm, confirmed with findings from venography, are added to seven previously reported cases revealed in the authors' search of the English-language literature. This rare anomaly usually shows as recurrent pulmonary emboli in patients with no underlying predisposition to deep venous thrombosis. Physical examination is usually not helpful in the diagnosis. Results of combined real-time and Doppler ultrasound should indicate the diagnosis, but venography is necessary for confirmation and further anatomic detail. Surgical treatment has been fraught with complications. Eight patients, including these two new cases, have undergone surgery, and none have had a recurrence of pulmonary embolism following surgery.


Eckol inhibits ultraviolet B-induced cell damage in human keratinocytes via a decrease in oxidative stress.
In previous reports, the antioxidant effects of eckol were shown to protect cells against hydrogen peroxide- and gamma ray-induced oxidative stress. In this study, the role of eckol in protecting human skin keratinocytes (HaCaT) against UVB-induced oxidative cell damage was investigated. Also, triphlorethol-A, one of the chemical components in Ecklonia cava, and quercetin a well known antioxidant, were compared with eckol in terms of antioxidant activity based on chemical structure. Eckol decreased UVB-induced intracellular reactive oxygen species (ROS), decreased injury to cellular components resulting from UVB-induced oxidative stress, and restored cell viability. In addition, eckol reduced UVB-induced apoptosis by inhibiting the disruption of mitochondrial membranes. These results suggest that eckol protects human keratinocytes against UVB-induced oxidative stress by scavenging ROS, thereby lessening injury to cellular components.


A nonenveloped virus with a lipid envelope: hepatitis A virus as used in virus-reduction studies.
Recently, a quasi-lipid-enveloped (LE) form of the traditionally nonlipid-enveloped (NLE) hepatitis A virus (HAV) was described in human serum and cell culture-derived HAV stocks. This discovery challenges the understanding of HAV reduction in virus clearance studies of plasma products, which were performed under the premise of an NLE nature of this virus. Here, the presence of LE particles in HAV stocks used for reduction studies was verified, and the hypothesis that LE and NLE particles might contribute to the differential heat sensitivity of HAV variants during heat treatment of human serum albumin was evaluated. Cell culture lysates and supernatants of two cytopathic HAV variants, HM175/18f and HM175/24a, were characterized for their LE and NLE particle content by isopycnic gradient centrifugation. The obtained fractions were characterized for relative infectivity and then subjected to heat treatment (58.0 ± 1.0°C for 590 ± 10 minutes) in 12.5% human serum albumin to investigate their respective heat sensitivity. Preparations of the two HAV variants contained either LE particles (HM175/24a) or LE and NLE particles (HM175/18f) with equivalent specific infectivity. For HM175/18f, heat sensitivity of LE and NLE fractions did not differ significantly, and inactivation of the whole virus stock was identical to the NLE particle inactivation profile, whereas the HM175/24a variant was more heat sensitive. The results indicate that, in heat-treatment studies, the LE or NLE HAV phenotype is less important than the choice of HAV variant, and the most heat-resistant HM175/18f should be used.


[Gene mutations from 511 myelodysplastic syndromes patients performed by targeted gene sequencing].
Objective: To study the characteristics of gene mutations in Chinese myelodysplastic syndromes (MDS) patients. Methods: A total of 511 Chinese patients with MDS performed 112-gene targeted sequencing were retrospectively analyzed. Results: Eighty-three distinct mutant genes were found in 511 patients with MDS. Amongst these, the most frequent mutations was associated with epigenetics (50%) , followed by spliceosome (37%) , signal transduction (34%) , transcription factors (24%) and cell cycle/apoptosis (17%) . 439 subjects (86%) had at least one gene mutation. The mean number of mutations in refractory anemia with unilineage dysplasia (RCUD) was 1.25, refractory anemia with multilineage dysplasia (RCMD) was 1.73, refractory anemia with ring sideroblasts (RARS) was 2.79, refractory anemia with excess blasts-1 (RAEB-1) was 2.22, RAEB-2 was 2.34, MDS with isolated 5q- was 2.67, MDS, unclassified (MDS-U) was 2.00. U2AF1 mutant subjects were more likely to have isolated+8[Q<0.001, OR=4.42 (95% CI 2.23-8.68) ]and less likely to have complex karyotypes[Q=0.005, OR=0.22 (95% CI 0.04-0.72) ]. According to the number of gene mutations, all subjects were categorized into three groups, namely group with 0-1 mutation, with 2 mutations and with three or more mutations. There was a significant difference in overall survival (OS) among three groups (P=0.041) . Conclusion: About 90% patients with MDS have at least one gene mutation. Genes associated with epigenetics and spliceosome are most common mutated genes in MDS. The increased numbers of gene mutations accompany with disease evolution and associate with poor prognosis.


Quantitative mRNA Imaging with Dual Channel qFIT Probes to Monitor Distribution and Degree of Hybridization.
Fluorogenic oligonucleotide probes facilitate the detection and localization of RNA targets within cells. However, quantitative measurements of mRNA abundance are difficult when fluorescence signaling is based on intensity changes because a high concentration of unbound probes cannot be distinguished from a low concentration of target-bound probes. Here, we introduce qFIT (quantitative forced intercalation) probes that allow the detection both of probe-target complexes and of unbound probes on separate, independent channels. A surrogate nucleobase based on thiazole orange (TO) probes the hybridization status. The second channel involves a nonresponsive near-IR dye, which serves as a reporter of concentration. We show that the undesirable perturbation of the hybridization reporter TO is avoided when the near-IR dye Cy7 is connected by means of short triazole linkages in an ≥18 nucleotides distance. We used the qFIT probes to localize and quantify oskar mRNA in fixed egg chambers of wild-type and mutant Drosophila melanogaster by wash-free fluorescence in situ hybridization. The measurements revealed a relative 400-fold enrichment of oskar within a 3000 μm3 large volume at the posterior pole of stage 8-9 oocytes, which peaked at a remarkably high 1.8 μM local concentration inside 0.075 μm3 volume units. We discuss detection limits and show that the number of oskar mRNA molecules per oocyte is independent of the oocyte size, which suggests that the final levels are attained already during the onset of oskar localization at stage 8.


Optimization of CRISPR/Cas9 genome editing in cotton by improved sgRNA expression.
When developing CRISPR/Cas9 systems for crops, it is crucial to invest time characterizing the genome editing efficiency of the CRISPR/Cas9 cassettes, especially if the transformation system is difficult or time-consuming. Cotton is an important crop for the production of fiber, oil, and biofuel. However, the cotton stable transformation is usually performed using Agrobacterium tumefaciens taking between 8 and 12 months to generate T0 plants. Furthermore, cotton is a heterotetraploid and targeted mutagenesis is considered to be difficult as many genes are duplicated in this complex genome. The application of CRISPR/Cas9 in cotton is severely hampered by the long and technically challenging genetic transformation process, making it imperative to maximize its efficiency. In this study, we provide a new system to evaluate and validate the efficiency of CRISPR/Cas9 cassettes in cotton using a transient expression system. By using this system, we could select the most effective CRISPR/Cas9 cassettes before the stable transformation. We have also optimized the existing cotton CRISPR/Cas9 system to achieve vastly improved mutagenesis efficiency by incorporating an endogenous GhU6 promoter that increases sgRNA expression levels over the Arabidopsis AtU6-29 promoter. The 300 bp GhU6.3 promoter was cloned and validated using the transient expression system. When sgRNAs were expressed under the control of the GhU6.3 promoter in CRISPR/Cas9 cassettes, expression levels were 6-7 times higher than those provided by the AtU6-29 promoter and CRISPR/Cas9-mediated mutation efficiency was improved 4-6 times. This study provides essential improvements to maximize CRISPR/Cas9-mediated mutation efficiency by reducing risk and workload for the application of CRISPR/Cas9 approaches in the targeted mutagenesis of cotton.


Proteins and chemical chaperones involved in neuronal nicotinic receptor expression and function: an update.
Neuronal nicotinic ACh receptors (nAChRs) are a family of ACh-gated cation channels, and their homeostasis or proteostasis is essential for the correct physiology of the central and peripheral nervous systems. The proteostasis network regulates the folding, assembly, degradation and trafficking of nAChRs in order to ensure their efficient and functional expression at the cell surface. However, as nAChRs are multi-subunit, multi-span, integral membrane proteins, the folding and assembly is a very inefficient process, and only a small proportion of subunits can form functional pentamers. Moreover, the efficiency of assembly and trafficking varies widely depending on the nAChR subtypes and the cell type in which they are expressed. A detailed understanding of the mechanisms that regulate the functional expression of nAChRs in neurons and non-neuronal cells is therefore important. The purpose of this short review is to describe more recent findings concerning the chaperone proteins and target-specific and target-nonspecific pharmacological chaperones that modulate the expression of nAChR subtypes, and the possible mechanisms that underlie the dynamic changes of cell surface nAChRs. This article is part of a themed section on Nicotinic Acetylcholine Receptors. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v175.11/issuetoc.


Identification of the DNA sequence required for transposition immunity of the gamma delta sequence.
A plasmid containing the transposon gamma delta sequence was immune to further insertion of gamma delta (transposition immunity). Plasmids carrying a fragment containing either 0.2 kilobase pairs of the gamma end or 0.4 kilobase pairs of the delta end of the gamma delta sequence were immune, and other parts of the gamma delta sequence did not confer immunity. The terminal 38-base-pair (bp) sequence of the delta end of the gamma delta was sufficient to confer immunity, the 38-bp sequence of the gamma end conferred only moderate immunity, and the terminal 35-bp sequence, which was completely identical at both the gamma and delta ends, was insufficient to confer immunity.


Cerium ions in the histochemical demonstration of second-messenger enzymes.
Second-messenger systems are involved in the regulation of numerous cellular processes. Adenylate cyclase (AC) and guanylate cyclase (GC) enzymes are in key positions in the regulation of these systems. The cerium method has been successfully applied to demonstrate amine- and neuropeptide-stimulated AC in rat nervous and adipose tissues and human sweat glands at the electron microscopic level. AC was also localized in cultured neurons. Nitric oxide compounds stimulated GC were demonstrated in rat hippocampal areas. Enzyme reactions were located in neurons pre- and postsynaptically in synapses; in addition, GC activity was seen intraneuronally and in glial cells. Adipocytes and eccrine glandular cells exhibited reaction products in their plasmalemmas. Optimal histochemical conditions are described, combined with control experiments. Some handicaps, related to the sensitivity of the enzymes to the fixatives, penetration problems of cerium salts, and especially the specificity of the method in phosphatase enzyme histochemistry in general are discussed.


Delayed presentation of a virilising, pure testosterone-secreting adrenocortical carcinoma with coexistent composite myelolipoma and a venous thrombus extending to the heart.
A 40-year-old normotensive woman presented with abnormal facial hair for 4 years and amenorrhoea for 13 years. Hormonal, biochemical and haematological evaluation showed isolated elevation of serum testosterone and free testosterone. Her follicle-stimulating hormone and luteinising hormone were in the premenopausal range. Until recently she had reconciled to early 'menopause' and visited beauty clinics but never sought medical evaluation. Imaging revealed an enhancing left adrenal mass with fat densities and venous thrombus extending through the inferior vena cava to a 7 cm mass in the right atrium. She underwent left kidney-preserving surgery utilising hypothermic cardiopulmonary bypass with early clamping of the pulmonary artery without circulatory arrest. Histology showed adrenocortical carcinoma with composite incidental myelolipoma and neoplastic thrombus. At 2 months, testosterone has normalised and she is doing well. Isolated testosterone-secreting adrenocortical carcinoma with massive venous thrombus is rare as is coincidental composite macroscopic myelolipoma.


Proteomics: technologies and applications.
Enabling technologies for proteomics (studies examining the protein complement of the genome) have been in development for over 20 years. However, more recently, the field has become formalized by combining the techniques for large-scale protein separation (two-dimensional electrophoresis) with very precise, high fidelity approaches to the analysis and characterisation of the separated proteins (mass spectrometry). These approaches bring to reality the powerful scope of proteomics, enabling the interpretation of cell physiology through profiling the molecular biodescriptors of gene expression, the proteins. Applications of proteomics are wide ranging and can be found within many disciplines, although one of the more developed fields that is examined here covers the arena of toxicological and pharmacological profiling. In this paper, the current technologies that make proteomics possible will be looked at. Some areas where proteomics has delivered valuable biological insight will also be considered, and there will be a look towards future areas of development.


Evaluation of prostatic carcinoma: noninvasive and preoperative techniques.
Transrectal ultrasonography was performed in 345 patients with clinical benign prostatic hyperplasia who underwent simple prostatectomy. Four patients with negative ultrasonic scans were found to have stage A2 prostatic tumors. Of the 57 patients with positive scans, ten were found to have clinically unsuspected carcinoma; however, changes secondary to carcinoma could not reliably be differentiated from those resulting from chronic inflammation and calculi. Transrectal portable ultrasonography is useful for the staging of patients with prostatic cancer, and four of the ten patients with seemingly stage B disease were found to have seminal vesicle involvement. Tumor response to endocrine therapy can be objectively monitored, but tumor reactivation often goes undetected. Because the technique does not reliably differentiate carcinoma, chronic inflammation, and calculi, the use of transrectal ultrasonography in the screening of asymptomatic patients in an effort to detect early carcinoma appears to be of limited value.


Synergic Adsorption-Biodegradation by an Advanced Carrier for Enhanced Removal of High-Strength Nitrogen and Refractory Organics.
Coking wastewater contains not only high-strength nitrogen but also toxic biorefractory organics. This study presents simultaneous removal of high-strength quinoline, carbon, and ammonium in coking wastewater by immobilized bacterial communities composed of a heterotrophic strain Pseudomonas sp. QG6 (hereafter referred as QG6), ammonia-oxidizing bacteria (AOB), and anaerobic ammonium oxidation bacteria (anammox). The bacterial immobilization was implemented with the help of a self-designed porous cubic carrier that created structured microenvironments including an inner layer adapted for anaerobic bacteria, a middle layer suitable for coaggregation of certain aerobic and anaerobic bacteria, and an outer layer for heterotrophic bacteria. By coating functional polyurethane foam (FPUF) with iron oxide nanoparticles (IONPs), the biocarrier (IONPs-FPUF) could provide a good outer-layer barrier for absorption and selective treatment of aromatic compounds by QG6, offer a conducive environment for anammox in the inner layer, and provide a mutualistic environment for AOB in the middle layer. Consequently, simultaneous nitrification and denitrification were reached with the significant removal of up to 322 mg L-1 (98%) NH4, 311 mg L-1 (99%) NO2, and 633 mg L-1 (97%) total nitrogen (8 mg L-1 averaged NO3 concentration was recorded in the effluent), accompanied by an efficient removal of chemical oxygen demand by 3286 mg L-1 (98%) and 350 mg L-1 (100%) quinoline. This study provides an alternative way to promote synergic adsorption and biodegradation with the help of a modified biocarrier that has great potential for treatment of wastewater containing high-strength carbon, toxic organic pollutants, and nitrogen.


Sub-threshold stimulation in variants of atrioventricular nodal re-entrant tachycardia: electrophysiological effects and impact for guidance of slow pathway ablation.
Sub-threshold stimulation (STS) applied during atrioventricular nodal re-entrant tachycardia (AVNRT) of the common (slow-fast) type has been shown to effectively characterise target sites suitable for slow pathway (SP) ablation but has not been investigated in the setting of fast-slow and slow-slow variants. Seventeen consecutive patients (52+/-16 years, 12 female) with sustained uncommon type AVNRT (fast-slow: n = 13, slow-slow: n = 4) were investigated. Mapping of the SP was started postero-septally close to the coronary sinus ostium and continued toward mid-septal sites, if required. Target sites for STS were selected according to established criteria including the recording of the earliest retrograde atrial activation during AVNRT. Long duration (5 s) constant current STS during AVNRT variants was performed in a stepwise manner (max 5 mA) at each site eligible for SP ablation until termination or capture occurred. Radiofrequency current (RFC) was delivered following successful STS termination of tachycardia (65 degrees C, 60 s) and exclusion of catheter dislodgement. Uncommon AVNRT with a mean cycle length of 405+/-70 ms was induced without spontaneous termination in all patients. Interruption of AVNRT variants due to selective STS-induced block of the retrograde (n = 12) or anterograde (n =2) SP occurred without capture in 14/17 (82%) patients. This was exclusively observed at sites with successful subsequent RFC application. AVNRT was rendered non-inducible in all patients after a median of 1 (1-11) RFC pulses without complications. Uncommon AVNRT can be interrupted by STS delivered at subsequently successful target sites for SP ablation in most patients (82%). The high positive and negative concordance between the effects of STS and following RFC application indicates that STS-mapping is also useful in the setting of AVNRT variants.


Probing the Interaction of Heavy and Transition Metal Ions with Silver Nanoparticles Decorated on Graphene Quantum Dots by Spectroscopic and Microscopic Methods.
We report a comprehensive study of the interaction of transition and heavy metal ions with graphene quantum dots-capped silver nanoparticles (AgGQDs) using different spectroscopic and microscopic techniques. High-resolution transmission electron microscopy studies show that the interaction of metal ions with AgGQDs leads to the formation of metal oxides, the formation of zerovalent metals, and the aggregation of Ag nanoparticles (AgNPs). The metal ions may interact with AgGQDs through selective coordination with -OH and -COOH functionalities, adsorption on the graphene moiety, and directly to AgNPs. For instance, the interaction of Cd2+ with AgGQDs altered the spherical shape of AgNPs into a chain-like structure. On the contrary, the formation of PbO is observed after the addition of Pb2+ to AgGQDs. Interestingly, the interaction of AgGQDs with Hg2+ results in the complete dissolution of Ag0 from the surface of GQDs and subsequent deposition of Hg0 on the graphene moiety of GQDs. Unlike transition metal ions, Cd2+, Pb2+, and Hg2+ can adsorb strongly on the graphene surface at the bridge, hollow, and top sites, respectively. This special interaction of heavy metal ions with the graphene surface would decide the mechanistic pathway in which the reaction proceeds. The transition metal ions Cu2+, Zn2+, Co3+, Mn2+, Ni2+, and Fe3+ induced the aggregation of AgNPs.


Office evaluation of bone tumors.
A familiarity with the characteristics and behavior of musculoskeletal neoplasms will allow for the delivery of timely and appropriate treatment. In this article the significance of historical and clinical data is discussed in general terms, followed by a rationale for office-based evaluation of these lesions. Some of the more common lesions are then discussed in greater detail.


Hashimoto's thyroiditis associated with urticaria and angio-oedema: disappearance of cutaneous and mucosal manifestations after thyroidectomy.
A 60 year old woman affected by Hashimoto's thyroiditis presented with a history of recurring episodes of urticaria and angio-oedema. Clinical and laboratory evaluation of the patient excluded allergy to external agents, hereditary angio-oedema, and occult infections. A pathogenic relation between Hashimoto's thyroiditis and chronic urticaria/angio-oedema was suspected. However, treatment with L-thyroxine had no influence on the frequency and severity of the cutaneous and mucosal manifestations, which occurred almost daily and required repeated administration of steroids. The patient therefore underwent total thyroidectomy. Cytometric analysis of intrathyroidal lymphocyte subsets showed unusual abnormalities. Urticaria and angio-oedema completely remitted after surgery; 18 months postoperatively the patient was still asymptomatic.


Distinct intracellular lysozyme content in normal granulocytes and monocytes: a quantitative immunoperoxidase and ultrastructural immunogold study.
Using two different immunological methods, we performed a quantitative estimation of lysozyme (LZ) in normal mature granulocytes and monocytes. An immunoperoxidase reaction for LZ in granulocytes and monocytes of 10 healthy donors measured by a scanning microdensitometer as arbitrary units showed a significantly higher LZ content in granulocytes than in monocytes. An ultrastructural immunogold reaction (IGR) for LZ performed on post-embedded thin sections showed a higher number of total gold grains in neutrophilic granulocytes than in monocytes. In monocytic granules we found a high density of gold grains per micron 2, whereas in granulocytic granules lower values were obtained. In granulocytes, LZ was found in both primary myeloperoxidase (MPO)-positive and secondary MPO-negative granules, and in monocytes the granules showed weak MPO reactivity and high LZ content. Granulocytes previously subjected to phagocytosis of bacteria and latex particles showed release of LZ on degranulation inside the phagosome, whereas in monocytes the granules remained outside the phagosome and released LZ without degranulation. Our study demonstrated a significantly higher total LZ content in granulocytes, a higher granular LZ content in monocytes, and release of LZ from intact monocyte granules during phagocytosis.


Mid-term results of a modified arterial switch operation using the direct reconstruction technique of the pulmonary artery.
There is ongoing discussion as to whether it is beneficial to avoid pulmonary sinus augmentation in the arterial switch operation. We report a single-surgeon series of mid-term results for direct pulmonary artery anastomosis during switch operation for transposition of the great arteries (TGA). This retrospective study includes 17 patients with TGA, combined with an atrial septal defect, patent foramen ovale or ventricular septal defect. Patient data was analyzed from hospital charts, including operative reports, post-operative course, and regular follow-up investigations. The protocol included cardiological examination by a single pediatric cardiologist. Echocardiographic examinations were performed immediately after arrival on the intensive unit, before discharge, and then after three, six, and 12 months, followed by yearly intervals. Pulmonary artery stenosis (PAS) was categorized into three groups according to the Doppler-measured pulmonary gradient: grade I (trivial stenosis) = increased pulmonary flow with a gradient below 25 mm Hg; grade II (moderate stenosis) = a gradient ranging from 25 to 49 mm Hg; and grade III (severe stenosis) = a gradient above 50 mm Hg. Follow-up data was available for all patients. The length of follow-up ranged from 1.2 to 9.7 years, median: 7.5 years (mean 6.1 years ± 14 months). During follow-up, 12 patients (70.6%) had no (or only trivial) PAS, five patients (29.4%) had moderate stenosis without progress, and no patient had severe PAS. Cardiac catheterization after arterial switch operation was performed in 11 patients (64.7%) and showed a good correlation with echocardiographic findings. During follow-up there was no reintervention for PAS. Direct reconstruction of the neo-pulmonary artery is a good option in TGA with antero-posterior position of the great vessels, with very satisfactory mid-term results.


[Study on the release and mechanism of carbon nanotubes loaded with verapamil hydrochloride in vitro].
The aim of this study was to load Verapamil Hydrochloride to carboxylated multi-walled carbon nanotubes( c-CNTs) and discuss the mechanism of drug release which could act as an effective basis for c-MWNTs used as drug carriers of controlled and sustained release delivery system. Raw CNTs were treated with mixed strong acid to obtain c-CNTs. Raman, IR, SEM and HR-TEM were used to characterize the CNTs and investigate the loading sites for drugs. The release behavior of the drug delivery system in vitro and the release model were studied. The raw CNTs were successfully grafted with carboxyl group by acid treatment. The water-soluble ability of c-CNTs was greatly improved. The length of c-CNTs was 200-300nm. Meanwhile, the ends of c-CNTs were opened. The results of the drug loading experiment showed that the more adding drugs, the larger loading content of drugs. Most of the drugs were loaded into the inner pores of c-CNTs when adding drugs was no more than 0.1 as quantity as c-CNTs. As the quantity of adding drugs increased, the drugs were loaded both in the inner pores and on the out-wall of c-CNTs. The release results in vitro showed release mechanism had something with the quantity of adding drugs. C-CNTs can be used as carriers of sustained and controlled release delivery system. Ideal release behavior of drugs can be achieved by choosing appropriate formula.


Comprehensive Proteome Reveals the Key Lethal Toxins in the Venom of Jellyfish Nemopilema nomurai.
Jellyfish stings are a major threat to human beings in coastal areas of the world. Each year, hundreds of thousands of victims are stung by venomous jellyfish. Nemopilema nomurai is a dangerous species with a large number of victims including many deaths. N. nomurai venom is a complex cocktail that is rich in proteins and peptides, and it is secreted by nematocysts for prey or defense. Previous studies have identified hundreds of toxins in the venom of N. nomurai; however, it is unclear which toxin(s) is responsible for lethality. Herein, we isolated the lethal fraction (NnLF) from N. nomurai venom with multiple chromatography. NnLF showed strong lethality to mice, and the toxicology results were consistent with the clinical symptoms of dead patients after N. nomurai sting, which indicated that NnLF contained the key lethal toxins in the venom. Subsequently, proteomic analysis was performed to identify the toxins in NnLF, and a total of 13 toxin homologues were identified, including phospholipase, potassium channel inhibitor, hemolysin, thrombin, etc. Moreover, in vitro toxicity assays further verified the phospholipase A2 and hemolytic activity of NnLF. These results revealed that cell membrane-targeted toxins, including channel-forming toxins, potassium channel inhibitors, and especially phospholipases, played very important roles in the lethality of N. nomurai sting. Moreover, blood toxins such as thrombin-like toxin and hemolysins might be synergistically involved in lethality. These findings advance the understanding of lethality caused by N. nomurai sting and will be significant for the development of drugs to treat this jellyfish sting in the future.


PCR-based detection of Angiostrongylus cantonensis in tissue and mucus secretions from molluscan hosts.
Angiostrongylus cantonensis is a common cause of human eosinophilic meningitis. Recent outbreaks of this infection have shown that there is a need to determine the distribution of this nematode in the environment in order to control transmission. A. cantonensis is generally identified morphologically in the molluscan intermediate host by microscopic examination, which can be labor-intensive. The aim of this study was to develop a PCR-based method to detect A. cantonensis directly from molluscan tissue. A total of 34 Parmarion cf. martensi (Simroth) semislugs, 25 of which were naturally infected with A. cantonensis, were used to develop this assay. Tissue pieces (approximately 25 mg) were digested with pepsin-HCl to recover third-stage larvae for morphological identification or were used for DNA extraction. PCR primers were designed to amplify 1,134 bp from the Angiostrongylus 18S rRNA gene, and the amplicons produced were sequenced for identification at the species level. Both microscopy and the PCR-DNA sequencing analysis indicated that the same 25 semislugs were positive for A. cantonensis, showing that the two methods were equally sensitive and specific for this application. However, morphological detection requires access to living mollusks, whereas molecular analysis can also be performed with frozen tissue. The PCR-DNA sequencing method was further evaluated using tissue from Veronicella cubensis (Pfeiffer) slugs and mucus secretions from infected P. martensi. To our knowledge, this is the first use of a PCR-based method to confirm the presence of A. cantonensis in mollusks collected in the environment.


EEG-fMRI: Ballistocardiogram Artifact Reduction by Surrogate Method for Improved Source Localization.
For the analysis of simultaneous EEG-fMRI recordings, it is vital to use effective artifact removal tools. This applies in particular to the ballistocardiogram (BCG) artifact which is difficult to remove without distorting signals of interest related to brain activity. Here, we documented the use of surrogate source models to separate the artifact-related signals from brain signals with minimal distortion of the brain activity of interest. The artifact topographies used for surrogate separation were created automatically using principal components analysis (PCA-S) or by manual selection of artifact components utilizing independent components analysis (ICA-S). Using real resting-state data from 55 subjects superimposed with simulated auditory evoked potentials (AEP), both approaches were compared with three established BCG artifact removal methods: Blind Source Separation (BSS), Optimal Basis Set (OBS), and a mixture of both (OBS-ICA). Each method was evaluated for its applicability for ERP and source analysis using the following criteria: the number of events surviving artifact threshold scans, signal-to-noise ratio (SNR), error of source localization, and signal variance explained by the dipolar model. Using these criteria, PCA-S and ICA-S fared best overall, with highly significant differences to the established methods, especially in source localization. The PCA-S approach was also applied to a single subject Berger experiment performed in the MRI scanner. Overall, the removal of BCG artifacts by the surrogate methods provides a substantial improvement for the analysis of simultaneous EEG-fMRI data compared to the established methods.


Family Sense-Making After a Down Syndrome Diagnosis.
The script of parenting shifts when parents learn of their child's Down syndrome diagnosis. To build a theory of the diagnostic experience and early family sense-making process, we interviewed 33 parents and nine grandparents living in the United States who learned prenatally or neonatally of their child's diagnosis. The core category of rescuing hope for the future encompassed the social process of sense-making over time as parents managed their sorrow, shock, and grief and amassed meaningful messages that anchored them as they looked toward the future. Application of the theory to practice underscores the import of early professional support offered to parents at key points in the sense-making process: Early as they disclose the news of the diagnosis to family and friends, and later close friends and kin assimilate meaningful messages about what the diagnosis means as they recalibrate expectations for a hopeful future.


From Neonatal Intensive Care to Neurocritical Care: Is It Still a Mirage? The Sicilian Multicenter Project.
Neonatal brain injury (NBI) can lead to a significant neurological disability or even death. After decades of intense efforts to improve neonatal intensive care and survival of critically ill newborns, the focus today is an improved long-term neurological outcome through brain-focused care. The goal of neuroprotection in the neonatal intensive care unit (NICU) is the prevention of new or worsening NBI in premature and term newborns. As a result, the neonatal neurocritical care unit (NNCU) has been emerging as a model of care to decrease NBI and improve the long-term neurodevelopment in critically ill neonates. Neurocritical care (NCC) Sicilian project includes three academic sites with NICU in Sicily (Catania, Messina, and Palermo), and its primary goal is to develop neurocritical neonatal care unit (NNCU). In 2018, the three NICUs created a dedicated space for neonates with primary neurological diagnosis or at risk for neurological injuries-NNCU. Admission criteria for eligible patients and treatment protocols were created. Contact with parents, environmental protection, basic monitoring, brain monitoring, pharmacological therapy, and organization of the staff were protocolized. Evaluation of the efforts to establish NNCU within existing NICU, current protocols, and encountered problems are shown. Implications for Practice. Our outcome confirmed the need for dedicated NNCU for neuroprotection of critically ill neonates at risk for a neurological injury. Although the literature on neonatal neurocritical care is still scarce, we see the value of such targeted approach to newborn brain protection and therefore we will continue developing our NNCU, even though there have been problems encountered. The project of building NNCU will continue to be closely monitored. The development of our neonatal neurocritical model of care is far from being completed. Although it is currently limited to the Sicilian area only, the goal of this paper is to share the development of this multicenter interdisciplinary project focused on a newborn brain protection. After evaluating our outcome, we strongly believe that a combined expertise in neonatal neurology and neonatal critical care can lead to an improved neurodevelopmental outcome for critically ill neonates, from the extremely preterm to those with brain injuries.


[Genetic evaluation for rifampicin-susceptibility of Mycobacterium tuberculosis].
We evaluated the relationship between rifampicin (RFP)-susceptibility and genetic alterations in rpoB gene of clinically isolated Mycobacterium tuberculosis strains, collected from different geographic locations within Japan. Alterations in rpoB gene were detected by PCR-direct sequencing analysis, and the minimum inhibitory concentrations (MICs) of RFP were determined by broth microdilution method using Middlebrook 7H9 broth. One hundred and sixty six epidemiologically unrelated M. tuberculosis strains were examined. A total of 99 genetic alterations in the 69 bp core region of rpoB gene were detected in 95 out of 166 strains. Among them, 96 out of 166 strains showed RFP-resistant phenotypes with MICs > or = 2 micrograms/ml. Examination of the correlation between the MICs of RFP and amino acid substitutions in 69 bp core region of rpoB gene revealed that 58 out of 59 strains containing amino acid substitution in codon 531 showed highly RFP-resistant phenotypes with MICs > or = 64 micrograms/ml. In contrast, a variable level of RFP-susceptibility was observed among strains containing amino acid substitutions in either codon 516 or 526. On the other hand, the MICs of three strains with a point mutation in either codon 515 or 533 were all < or = 1 microgram/ml. Our results suggest that rpoB gene sequencing is useful for not only the detection of RFP-resistant M. tuberculosis strains, but also the prediction of RFP-susceptibility of the strains.


Genetic analysis and characterization of a new maize association mapping panel for quantitative trait loci dissection.
Association mapping based on the linkage disequilibrium provides a promising tool to identify genes responsible for quantitative variations underlying complex traits. Presented here is a maize association mapping panel consisting of 155 inbred lines with mainly temperate germplasm, which was phenotyped for 34 traits and genotyped using 82 SSRs and 1,536 SNPs. Abundant phenotypic and genetic diversities were observed within the panel based on the phenotypic and genotypic analysis. A model-based analysis using 82 SSRs assigned all inbred lines to two groups with eight subgroups. The relative kinship matrix was calculated using 884 SNPs with minor allele frequency > or = 20% indicating that no or weak relationships were identified for most individual pairs. Three traits (total tocopherol content in maize kernel, plant height and kernel length) and 1,414 SNPs with missing data < 20% were used to evaluate the performance of four models for association mapping analysis. For all traits, the model controlling relative kinship (K) performed better than the model controlling population structure (Q), and similarly to the model controlling both population structure and relative kinship (Q + K) in this panel. Our results suggest this maize panel can be used for association mapping analysis targeting multiple agronomic and quality traits with optimal association model.


A prospective study of villous capillary lesions in complicated pregnancies.
The vascularity of placental tissue is dependent on various factors of which fetomaternal hypoxia plays a major role. Hypoxia can be of different types and each type influences the vascularity of the villi, especially terminal villi, in its own way. In this study, we attempted to identify villous vascular changes in a group of term placentae from mothers with diseases complicating pregnancy. Chorangiosis was the most frequently identified lesion while chorangioma was found in only 2 cases. There were no cases of chorangiomatosis. A few cases had normal villous vasculature. Maternal diseases have a major role in disrupting the placental vasculogenesis and angiogenesis by creating a hypoxic environment that may affect the fetus adversely. Hence, such conditions need to be identified early in pregnancy and managed appropriately as it is possible to maintain a normal vasculature and prevent neonatal mortality and morbidity if prompt intervention is done.


Turkish social attitudes towards to cancer prevention: a health belief model study.
This research was planned to reveal society's attitude towards cancer and early diagnosis using the health belief model. This study was planned as descriptive research in Isparta. A random sample of n=256 individuals of both genders was recruited at the largest shopping center. As a means of collection tool, a survey consisted of two forms, the first designed for sociodemographic information and the second covering 29 questions suitable for the content of Health Belief Model. Of the participants, 66.8% were female and 33.2% were male, and the average age was 33.3±11.0 years. Some 46.1% partly thought that they may develop cancer, and 49.6% were afraid of this possibility. As many as 50% indicated that cancer is an issue that comes from Allah. A significant difference was found between not going for control unless feeling bad, and blood analysis for cancer screening (χ2=3.780 p= 0.03). It was seen that in an area with a high rate of cancer, people's awareness of cancer prevention and early diagnosis and attitudes towards these are insufficient.


EGR1/LINC00839/SOX5 axis modulates migration, invasion and Gemcitabine resistance of bladder cancer cells.
Bladder cancer is one of the most common malignant tumors of the urinary system, and its incidence is increasing worldwide. However, the underlying mechanisms that trigger migration, invasion and chemotherapy resistance are unclear. Bioinformatics analysis of bladder cancer cohort indicated that LINC00839 is deregulated in bladder cancer. LINC00839 was validated and highly expressed in bladder cancer patients and cell lines. In addition, LINC00839 induced the migration, invasion and Gemcitabine resistance of bladder cancer cells. We identified that the transcription factor EGR1 directly repressed LINC00839 and thereby suppressed the migration and invasion of bladder cancer cells. Furthermore, LINC00839 interacted with miR-142, which subsequently regulated the expression of SOX5, a well-studied oncogene and targeted by miR-142. In addition, EGR1 served as a suppressive transcription factor of SOX5. Therefore, EGR1 directly or indirectly regulates SOX5 via LINC00839/miR-142 axis. LINC00839 induced Gemcitabine resistance by promoting autophagy. EGR1, LINC00839/miR-142 and SOX5 form a coherent feed-forward loop that modulates the migration, invasion and Gemcitabine resistance of bladder cancer.


Temperature compensated magnetic field sensing using dual S-bend structured optical fiber modal interferometer cascaded with fiber Bragg grating.
A temperature compensated magnetic field strength optical fiber sensor has been proposed and experimentally demonstrated. A fiber Bragg grating (FBG) is cascaded to modal interferometer (MI), which is fabricated by dual S-bend splicing between thin fiber (TF) and single mode fiber (SMF) with intentionally controlled misalignment between cores. We established a modified numerical model to describe the multi-mode interference of this exceptional S-bend and misalignment structure, together with the simulation based on beam propagation method to gain insight into its operation mechanism. The FBG is used to interrogate the temperature change, and then compensate the perturbation of temperature on transmission of the MI. Thanks to the proposed dual S-bend structure and the diameter-thinned TF used here; we have obtained high magnetic sensitivity of -0.0678 dB/Oe using only 4 mm TF after the elimination of ambient temperature change.


[Perioperative management in hemophilic arthropathy].
To evaluate the clinical results and improve perioperative management in patients with hemophilic arthropathy. From May 2002 to March 2007, 20 patients with hemophilic arthropathy were admitted. Fourteen patients (20 knees) were performed total knee arthroplasty, and 6 patients (8 hips) were performed total hip arthroplasty, while 2 patients were performed ankle arthrodesis. The average preoperative HSS score, hip Harris score and ankle AOFAS score were 83, 89 and 78 points respectively. All patients had been followed up, and the mean time was 27.3 months. The average postoperative knee HSS score increased to 83 points, and hip Harris score increased to 89 points, while the AOFAS score increased to 78 points. Delayed bleeding and deep venous thrombosis and late infection with loose prosthesis occurred in one case respectively. Arthroplasty and arthrodesis are promising for severe hemophilic arthropathy to obtain pain relief and functional improvement Correct perioperative management is the key to successful treatment in hemophilic arthropathy.