CD40L-stimulated B cells for ex-vivo expansion of polyspecific non-human primate regulatory T cells for translational studies.

The therapeutic applications of regulatory T cells (Tregs ) include treating autoimmune diseases, graft-versus-host disease and induction of transplantation tolerance. For ex-vivo expanded Tregs to be used in deceased donor transplantation, they must be able to suppress T cell responses to a broad range of human leukocyte antigen (HLA). Here, we present a novel approach for the expansion of polyspecific Tregs in cynomolgus macaques that was adapted from a good manufacturing practice-compliant protocol. Tregs were isolated by fluorescence-activated cell sorting (FACS) and expanded in the presence of a panel of CD40L-stimulated B cells (CD40L-sBc). Prior to Treg culture, CD40L-sBc were expanded in vitro from multiple major histocompatibility complex (MHC)-disparate macaques. Expanded Tregs expressed high levels of forkhead box protein 3 (FoxP3) and Helios, a high percentage of Treg -specific demethylated region (TSDR) demethylation and strong suppression of naïve T cell responses in vitro. In addition, these Tregs produced low levels of inflammatory cytokines and were able to expand post-cryopreservation. Specificity assays confirmed that these Tregs were suppressive upon activation by any antigen-presenting cells (APCs) whose MHC was shared by CD40L-sBc used during expansion, proving that they are polyspecific. We developed an approach for the expansion of highly suppressive cynomolgus macaque polyspecific Tregs through the use of a combination of CD40L-engineered B cells with the potential to be translated to clinical studies. To our knowledge, this is the first report that uses a pool of MHC-mismatched CD40L-sBc to create polyspecific Tregs suitable for use in deceased-donor transplants.

Ambulance use affects timely emergency treatment of acute ischaemic stroke.

INTRODUCTION: For acute ischaemic stroke patients, treatment with intravenous tissue plasminogen activator within a 4.5-hour therapeutic window is essential. We aimed to assess the time delays experienced by stroke patients arriving at the emergency department and to compare ambulance users and non-ambulance users. METHODS: We performed a prospective cohort study in a tertiary hospital in Hong Kong. All acute stroke patients attending the emergency department from January to June 2017 were recruited. Patients who were in hospital at the time of stroke onset and those who transferred from other hospitals were excluded. Three phases were compared between ambulance users and non-ambulance users: phase I, between stroke onset and calling for help; phase II, between calling for help and arriving at the emergency department; and phase III, between arriving and receiving medical assessment. RESULTS: Of 102 consecutive patients recruited, 48 (47%) patients arrived at the emergency department by ambulance. The percentage of stroke patients attending emergency department within the therapeutic window was significantly higher for ambulance users than for non-ambulance users (64.6% vs 29.6%; P<0.001). For phases I, II and III, the median times were significantly shorter for ambulance users (77.5, 32 and 8 min, respectively) than for non-ambulance users (720, 44.5 and 15 min, respectively; all P<0.001). CONCLUSION: Transport of patients to the emergency department by ambulance is important for timely and effective stroke treatment.

Predicting arboviral disease emergence using Bayesian networks: a case study of dengue virus in Western Australia.

A Bayesian Belief Network (BBN) for assessing the potential risk of dengue virus emergence and distribution in Western Australia (WA) is presented and used to identify possible hotspots of dengue outbreaks in summer and winter. The model assesses the probabilities of two kinds of events which must take place before an outbreak can occur: (1) introduction of the virus and mosquito vectors to places where human population densities are high; and (2) vector population growth rates as influenced by climatic factors. The results showed that if either Aedes aegypti or Ae. albopictus were to become established in WA, three centres in the northern part of the State (Kununurra, Fitzroy Crossing, Broome) would be at particular risk of experiencing an outbreak. The model can also be readily extended to predict the risk of introduction of other viruses carried by Aedes mosquitoes, such as yellow fever, chikungunya and Zika viruses.

Initial results of selective renal parenchymal clamping with an adjustable kidney clamp in nephron-sparing surgery: an easy way to minimise renal ischaemia.

INTRODUCTION: A renal parenchymal clamp has been used at our centre since March 2012. It is used in position over the kidney to achieve optimal vascular control of a tumour while minimising parenchymal ischaemia. This study aimed to report the feasibility, surgical outcome, and oncological control of a kidney clamp in partial nephrectomy. METHODS: This study was conducted at a teaching hospital in Hong Kong. Partial nephrectomies performed from January 2009 to March 2015 were reviewed. The tumour characteristics and surgical outcomes of kidney clamp were studied and compared with traditional hilar clamping. RESULTS: A total of 92 patients were identified during the study period. Kidney clamps were used in 20 patients and hilar clamping in 72, with a mean follow-up of 27 and 37 months, respectively. For patients in whom a kidney clamp was applied, all tumours were exophytic to a different extent and the majority (90%) were located at the polar region. The PADUA (preoperative aspects and dimensions used for an anatomical) classification nephrometry score was also lower than those in whom hilar clamping was used (7.07 vs 8.34; P=0.002). The clamp was used in open, laparoscopic, and robot-assisted surgery. Operating time was shorter (207 ± 72 mins vs 306 ± 80 mins; P<0.001) and estimated blood loss was lower (205 ± 191 mL vs 331 ± 275 mL; P=0.045) with kidney clamp. No acute kidney injury occurred. Postoperative renal function was comparable between the two groups. CONCLUSIONS: Partial nephrectomy using parenchymal clamping is safe and feasible in selected cases. The postoperative renal function and oncological control were satisfactory.

Bat coronavirus was detected positive from insectivorous bats in Krau Wildlife Reserve Forest.

Bats are flying mammals with unique immune systems that allow them to hold many pathogens. Hence, they are recognised as the reservoir of many zoonotic pathogens. In this study, we performed molecular detection to detect coronaviruses, paramyxoviruses, pteropine orthoreoviruses and dengue viruses from samples collected from insectivorous bats in Krau Reserve Forest. One faecal sample from Rhinolophus spp. was detected positive for coronavirus. Based on BLASTN, phylogenetic analysis and pairwise alignment-based sequence identity calculation, the detected bat coronavirus is most likely to be a bat betacoronavirus lineage slightly different from coronavirus from China, Philippines, Thailand and Luxembourg. In summary, continuous surveillance of bat virome should be encouraged, as Krau Reserve Forest reported a wide spectrum of biodiversity of insectivorous and fruit bats. Moreover, the usage of primers for the broad detection of viruses should be reconsidered because geographical variations might possibly affect the sensitivity of primers in a molecular approach.

Immunogenicity and safety profiles of genetic vaccines against human Her-2/neu in cynomolgus monkeys.

Her-2/neu is a well-characterized tumor-associated antigen, the overexpression of which in human carcinomas correlates with a poor prognosis. Here, we evaluated Her-2/neu-specific humoral and cellular immune responses in immunized monkeys after immunization with nonreplicating adenovirus (AdHM) expressing the extracellular and transmembrane domain of human Her-2/neu (HM) and/or naked DNA vaccine (pHM-hGM-CSF) expressing human granulocyte-macrophage colony-stimulating factor together with HM. Priming of monkeys with AdHM generated Her-2/neu-specific long-lasting antibody production. Furthermore, these Her-2/neu-specific antibodies produced by AdHM immunization, some of which shared epitope specificity with Herceptin, were able to induce antibody-dependent cellular cytotoxicity against Her-2-expressing target cells. Cellular immune responses were elicited in all monkeys immunized with Her-2/neu-expressing vaccine; interferon-gamma was secreted when these splenocytes were restimulated with Her-2/neu-expressing autologous cells, and immunization with AdHM induced Her-2/neu-specific lymphoproliferative responses. Further, immunization with pHM-hGM-CSF before AdHM immunization noticeably enhanced cytotoxic T-lymphocyte activity. In addition, we observed no abnormalities that would indicate that the genetic vaccines had toxic effects in the immunized monkeys. Thus, we can conclude that our genetic vaccines efficiently elicited Her-2/neu-specific humoral and cellular immune responses without causing severe adverse effects in nonhuman primates and that as such they warrant further clinical investigation.

A combination of rough-based feature selection and RBF neural network for classification using gene expression data.

This paper presents a novel rough-based feature selection method for gene expression data analysis. It can find the relevant features without requiring the number of clusters to be known a priori and identify the centers that approximate to the correct ones. In this paper, we attempt to introduce a prediction scheme that combines the rough-based feature selection method with radial basis function neural network. For further consider the effect of different feature selection methods and classifiers on this prediction process, we use the NaIve Bayes and linear support vector machine as classifiers, and compare the performance with other feature selection methods, including information gain and principle component analysis. We demonstrate the performance by several published datasets and the results show that our proposed method can achieve high classification accuracy rate.

Toxicity of repeated intravenous injection of gene therapeutics for X-CGD in mice.

We made gene therapeutics for X-chronic granulomatous disease (CGD) by transducing murine bone marrow-derived stem cells with MT-gp91 retrovirus and evaluated possible toxicity in mice as a prerequisite for human clinical trials. Male C57BL/6 mice were injected intravenously with gene therapeutics for X-CGD twice at an interval of two weeks at 5 x 10(7) cells/kg and sacrificed 2 weeks after the last administration. Significant changes noted in gene therapeutics for X-CGD-treated animals were an increase in white blood cell counts and a slight decrease in albumin/globulin ratio. The red pulp hyperplasia in the spleen accompanied with an increase in organ weight was considered to result from the accumulation of gene therapeutics for X-CGD, bone marrow-derived stem cells, in the spleen. No anti-gp91 antibody was detected in the sera collected from the animals treated with gene therapeutics for X-CGD. No integration of gp91 DNA from retroviral vector was detected in chromosomal DNA of gonads in animals dosed with the test substance, indicating no potential of genomic integration. In conclusion, the repeated dose of gene therapeutics for X-CGD exerted no toxicity. The splenic red pulp hyperplasia and the increase observed in white blood cell counts and in spleen weights were considered as pharmacological changes induced by the treatment.

Development of an E-learning System for the Endoscopic Diagnosis of Early Gastric Cancer: An International Multicenter Randomized Controlled Trial.

BACKGROUND: In many countries, gastric cancer is not diagnosed until an advanced stage. An Internet-based e-learning system to improve the ability of endoscopists to diagnose gastric cancer at an early stage was developed and was evaluated for its effectiveness. METHODS: The study was designed as a randomized controlled trial. After receiving a pre-test, participants were randomly allocated to either an e-learning or non-e-learning group. Only those in the e-learning group gained access to the e-learning system. Two months after the pre-test, both groups received a post-test. The primary endpoint was the difference between the two groups regarding the rate of improvement of their test results. FINDINGS: 515 endoscopists from 35 countries were assessed for eligibility, and 332 were enrolled in the study, with 166 allocated to each group. Of these, 151 participants in the e-learning group and 144 in the non-e-learning group were included in the analysis. The mean improvement rate (standard deviation) in the e-learning and non-e-learning groups was 1·24 (0·26) and 1·00 (0·16), respectively (P<0·001). INTERPRETATION: This global study clearly demonstrated the efficacy of an e-learning system to expand knowledge and provide invaluable experience regarding the endoscopic detection of early gastric cancer (R000012039).

Recurrent prostatic stromal tumour of uncertain malignant potential (STUMP) presenting with urinary retention 6 Years after transurethral resection of prostate (TURP).

CLINICAL PRESENTATION: A 56-year-old Chinese male with previously diagnosed prostatic stromal tumour of uncertain malignant potential (STUMP) presented with urinary retention 6 years after transurethral resection of prostate (TURP). TREATMENT AND OUTCOME: Cystoscopy showed a papillary tumour of the prostatic urethra causing near-complete obstruction. Repeat TURP was performed. He has been asymptomatic since. CONCLUSION: There has been fewer than 100 cases of this lesion reported worldwide. Definitive treatment is not well established. Longterm follow-up to monitor progression and possible recurrence is required, and repeat TURP or radical surgery may be necessary.

Induction of NG108-15 cells differentiation by human bone marrow stromal cells.

The survival and differentiation of neuronal cells is dependent on factors such as neurotrophins, cytokines and components of extracellular matrix. Bone marrow stromal cells have been shown to support the growth and differentiation of neuroblastoma cells. In an attempt to study the effects of bone marrow stromal cells on neuronal differentiation, we have co-cultured neuroblastoma x glioma hybrid NG108-15 cells with human bone marrow stromal cells. After co-culturing, clones exhibiting morphological differentiated phenotype and high level of neurofilament expression were isolated. Interestingly, these clones maintain their ability to proliferate in contrast to differentiated NG108-15 cells induced by dibutyryladenosine 3',5'-cyclic monophosphate. These results suggested that bone marrow stromal cells can induce partial differentiation of NG108-15 cells.

The effects of phorate and some of its metabolites incorporated in an artificial diet on liquid uptake by Aphis fabae scop.

[14C]Phorate and some of its metabolites were incorporated in an artificial diet and their effects on liquid uptake by Aphis fabae were studied. At sublethal and very low toxicity levels of phorate there was a tendency for a reduction in feeding with increasing concentrations. [14C]Phorate sulphone, [14C]phorate sulphoxide and [14C]phoratoxon sulphoxide at high concentrations gave very low mortality figures, but a significant drop in liquid uptake occurred with increasing concentrations. The results demonstrate that the presence of the insecticide rendered the diet less acceptable to the aphids which exhibited a wandering behaviour that might promote transmission of virus diseases.

Deltamethrin as a potential ovicidal pyrethroid against Plutella xylostella L.

The effect of deltamethrin on the hatching of eggs of two strains, susceptible (S) and resistant (R), of Plutella xylostella was investigated. Daily observations were made on the development of the embryos, hatching and larval mortality after treatment with different dosages of deltamethrin. This compound exhibited a dosage-mortality response against the S-strain eggs. The hatching of the R-strain eggs was also affected by deltamethrin but to a lesser extent. At least 50% of the S-strain larvae that hatched from the treated eggs died within a few hours, but the larvae of the R strain were more resistant. Seedlings treated with different dosages also showed a drop in egg hatching and less damage on the leaves. Deltamethrin had no effect on embryonic development.

Toxicity of deltamethrin and cypermethrin to the larvae of the diamond-back moth, Plutella xylostella L.

Two photostable pyrethroids, deltamethrin and cypermethrin, were tested against fourth instar larvae of a susceptible (S) strain and a resistant (R) strain of Plutella xylostella L. by topical application. Both compounds were very effective against the S strain larvae (LD50 for deltamethrin = 0.0014 micrograms/larva, LD50 for cypermethrin = 0.0046 micrograms/larva, at 48 h). However, the R strain collected from the field was greater than 1600-fold resistant to deltamethrin and greater than 30 000-fold resistant to cypermethrin. Deltamethrin was poorly synergised with piperonyl butoxide in the S strain, but much stronger synergism was obtained in the R strain. The possible reasons for such high levels of resistance and the potential use of synergised pyrethroids in Plutella control programmes are discussed.

The effects of removal and reconstruction of the anterior cruciate ligament on patellofemoral kinematics.

Patellofemoral pain may be associated with anterior cruciate ligament deficiency or may occur after anterior cruciate ligament reconstruction. We investigated the effects of the removal and reconstruction of the anterior cruciate ligament on the kinematics of the tibiofemoral and patellofemoral joints during physiologic levels of quadriceps muscle loads in seven cadaveric knees. A bone-patellar tendon-bone graft was used for intraarticular reconstruction of the anterior cruciate ligament. The spatial positions of the tibiofemoral and patellofemoral joints were measured between 0 degrees and 90 degrees of knee flexion in 15 degrees increments with a six degree-of-freedom digitizing system. Excision of the anterior cruciate ligament resulted in statistically significant increases in anterior tibial translation between 0 degrees and 90 degrees and valgus tibial rotation between 30 degrees and 90 degrees; intraarticular reconstruction returned these to levels not significantly different from those of the intact knee. Excision of the anterior cruciate ligament resulted in significant increases in lateral patellar tilt, ranging from 6.3 degrees to 9.0 degrees between full extension and 90 degrees of knee flexion, and in lateral patellar shift, ranging from 2.9 mm at 15 degrees of knee flexion to 5.9 mm at 90 degrees; intraarticular reconstruction returned these to levels not significantly different from those of the intact knee. Neither removal nor reconstruction of the anterior cruciate ligament significantly affected tibial internal-external rotation, patellar flexion, patellar mediolateral rotation, patellar anteroposterior translation, or patellar proximodistal translation.

STR data for the PowerPlex 16 loci for the Chinese population in Hong Kong.

Allele frequencies for the 15 STR loci included in the PowerPlex 16 kit were obtained from a sample of 247 unrelated Chinese in Hong Kong.

Structural modeling of differential diagnosis, treatment, and results for allergic rhinitis.

This paper analyzed the relationship among the differential diagnosis, treatment, and results for allergic rhinitis using the covariance structural model. The data were collected from 274 patients with suspected allergic rhinitis who visited the Otorlaryngology Department of the Paik Hospital during 1991-1993. After each patient's characteristics was categorized and combined into several common factors, covariance structure analysis was performed to analyze the structural relationships among the differential diagnosis, treatment, and results of treatment using the significant factors obtained from discriminant analysis. The significant characteristics influenced the diagnosis were the results of skin test from mite/animal, and from mugworts, the results from laboratory tests, rhinorrhea and sneezing, and nasal polyps. The significant characteristics that influenced the method of treatment were: nasal polyps, headache/general symptom, family history/medication, and septal deviation. Headache/general symptom was the only significantly influencing factor for the treatment results.

Data mining approach to policy analysis in a health insurance domain.

This study examined the characteristics of the knowledge discovery and data mining algorithms to demonstrate how they can be used to predict health outcomes and provide policy information for hypertension management using the Korea Medical Insurance Corporation database. Specifically, this study validated the predictive power of data mining algorithms by comparing the performance of logistic regression and two decision tree algorithms, CHIAD (Chi-squared Automatic Interaction Detection) and C5.0 (a variant of C4.5) using the test set of 4588 beneficiaries and the training set of 13,689 beneficiaries. Contrary to the previous study, the CHIAD algorithm performed better than the logistic regression in predicting hypertension, and C5.0 had the lowest predictive power. In addition, the CHIAD algorithm and the association rule also provided the segment-specific information for the risk factors and target group that may be used in a policy analysis for hypertension management.