Gate Keeper Research Articles

CAR PARKING SLOT DETECTION USING YOLO ALGORITHM

Finding free slots in the parking area has become a major problem that the modern society is facing nowadays, which also leads to traffic overcrowding at the parking area. As the number of vehicles are increasing at a quick pace find parking slots has become difficult. The proposed paper helps to solve the parking problem. In traditional parking system a person has to find vacant slot or a gate keeper needs to inform about the available parking slot at parking area. To address the issue, a system where the user will be able to know the number of vacant slots in a particular parking area along with total number of slots at the entrance of the parking area. As a critical component of this project, by built a car parking slot detection system which collects data from a camera at parking area. Today's most popular object detection model YOLO, is trained to achieve desired results with good precision scores at real-time speed. The model will be getting all the parking slots that are present in the parking area, to which marking boxes are drawn around for training. Real time processing is performed on the obtained data by the model to find whether the slot is vacant or occupied. Thus, it gives the information of available vacant slots to park the car at parking area. Key Words: YOLO, Object detection, Computer vision, Machine learning.

Human TP53 gene polymorphisms among patients with Hepatocellular carcinoma and chronic hepatitis B infection in Kenya

Background Human TP53 is the gatekeeper for generation of human cells and is highly conserved. Some alteration/mutation in TP53 adversely affects the regulatory function of the protein, potentially resulting in cancer. This study investigated mutations in codons 72 and 249 of TP53, among patients with hepatocellular carcinoma (HCC) and chronic hepatitis B virus (HBV) infection at the Moi Teaching and Referral Hospital (MTRH), Eldoret, Kenya. Methods In total, 33 HBV-positive patients attending MTRH hospital between September 2013 and July 2017 were purposely selected from medical records for the study; those with HCC were confirmed from the cancer registry. The patients were aged between 25-67 years, with a male-to-female ratio of 1.1:1. Blood samples were collected from the patients. DNA was extracted, amplified and sequenced using TP53 forward and reverse primers. Gene mutation detection and analysis was done on exons 4 codon 72 and exon 7 codon 249. Results Of the 33 patients, 75.8% were chronically infected with HBV and had HCC; the rest were HBsAg positive without HCC. Homozygous proline was prevalent (54.5%) at exon 4 codon 72, followed by heterozygous Arg/Pro (33.3%) and lastly homozygous Arg/Arg (12.1%). Pro/Pro allele was frequent in HCC group while Arg/Arg allele was common in patients without HCC. There was no significant association between the HCC and codon polymorphisms (P=0.12). In exon 7, codon 249, 24.2% of patients had an Arg/Ser mutation of which, 75.0% had HCC and 25.0% did not. There was no significant association between HCC patients and codon 249 mutation (P=0.15). Conclusion TP53 is a gene gate keeper, the mutations under study may dependently play a role in HCC development. This study did not find any association between TP53 mutations and presence of HCC. Therefore, TP53 Arg-72 and Ser-249 mutation is not a clear prognosis indicator for hepatocellular carcinoma among HBV infected patients in Kenya.

A184 HCPE CONTRIBUTES TO THE HOST INFLAMMATORY RESPONSE TO HELICOBACTER PYLORI

Abstract Background Helicobacter pylori (HP) colonizes chronically 50% of the world population, leading to gastric ulcers or cancers in 2-10 % of infections. Due to rising antibiotic resistance, curtailing HP-induced diseases requires the identification of new anti-HP molecules that can reduce HP viability or its ability to elicit inflammation. Amongst HP’s virulence factors are the Helicobacter Cysteine-rich Proteins (Hcp) that are secreted outside the bacteria and contain Sel-Like Repeats (SLR) which support protein / protein interactions involved in signal transduction across kingdoms. Several SLR-containing bacterial proteins including HcpA are involved in bacteria / host interactions. Thus, Hcps may affect HP’s virulence via interaction of their SLRs with host proteins. Aims We aim to elucidate the role of Hcps in HP’s virulence. Hcps were considered as immune decoys since anti-Hcp antibodies made in infected patients are unable to clear the infection. However, recent data suggest that Hcps interact with host components and may modulate the function of immune cells. However, their effects on gastric cells remain unknown and functional differences between Hcps remain to be explored. Methods This work uses knockout mutagenesis, interactions with gastric cells, transcriptomics, and microscopy. Results We focus on HcpE, the largest Hcp that may interact with many host proteins via its 9 SLRs motifs. We showed that HcpE is secreted outside HP in 2 lab strains and that ~ 30% of secreted HcpE was comprised in outer membrane vesicles that may deliver HcpE into eukaryotic cells by membrane fusion. We also showed that the DiSulfide Bond forming protein DsbK is essential for HcpE production and secretion. Here, we demonstrate the production and secretion of HcpE by clinical isolates in vitro, and its production in situ in infected patients. Using our HcpE and DsbK knockout mutants, we investigate the role of HcpE and other Hcps in interactions with human gastric cells in vitro and in ex vivo gastric explants, showing a role in inflammation activation. Finally, we demonstrate their role in murine gastric colonization and in murine splenocytes activation. Overall, our data show a role of Hcps in host / pathogen interactions largely due to immuno-modulatory functions and a role in gastric colonization, with a predominant role of HcpE over other Hcps. Conclusions The findings open up possibilities to use Hcps or the gate keeper DsbK that controls their production as novel therapeutic targets to alleviate the burden of HP-associated disease.This could benefit the ~6 million patients who develop gastric ulcers or cancers each year worldwide due to HP infection, also saving billions of annual health care costs. Funding Agencies Western University

Analysis of the neuromuscular deficits caused by STAM1 deficiency

The endosomal sorting complexes required for transport (ESCRT) pathway is composed of a series of protein complexes that are essential for sorting cargo through the endosome. In neurons, the ESCRT pathway is a key mediator of many cellular pathways that regulate neuronal morphogenesis as well as synaptic growth and function. The ESCRT-0 complex, consisting of HGS (hepatocyte growth factor-regulated tyrosine kinase substrate) and STAM (signal-transducing adaptor molecule), acts as a gate keeper to this pathway, ultimately determining the fate of the endosomal cargo. We previously showed that a single nucleotide substitution in Hgs results in structural and functional changes in the nervous system of teetering mice. To determine if these changes occurred as a function of HGS's role in the ESCRT pathway and its association with STAM1, we investigated if STAM1 deficiency also leads to a similar impairment of the nervous system. In contrast to teetering mice that die within 5 weeks of age and exhibit reduced body mass, 1-month-old Stam1 knockout mice were not visibly different from controls. However, by 3 months of age, STAM1 deficiency caused reduced muscle mass, strength, and motor performance. These changes in motor function did not correlate with either a loss in motor neuron number or abnormal myelination of peripheral nerves. Instead, the motor endplate structure was altered in the Stam1 knockout mice by 1 month of age and continued to degenerate over time, correlating with a significant reduction in muscle fiber size and increased expression of the embryonic γ acetylcholine receptor (AChR) subunit at 3 months of age. There was also a significant reduction in the levels of two presynaptic SNARE proteins, VTI1A and VAMP2, in the motor neurons of the Stam1 knockout mice. As loss of STAM1 expression replicates many of the structural changes at the motor endplates that we have previously reported with loss of HGS, these results suggest that the HGS/STAM1 complex plays a critical role in maintaining synaptic structure and function in the mammalian nervous system.

Glycoprotein 2 as a gut gate keeper for mucosal equilibrium between inflammation and immunity.

Glycoprotein 2 (GP2) is a widely distributed protein in the digestive tract, contributing to mucosal barrier maintenance, immune homeostasis, and antigen-specific immune response, while also being linked to inflammatory bowel disease (IBD) pathogenesis. This review sheds light on the extensive distribution of GP2 within the gastrointestinal tract and its intricate interplay with the immune system. Furthermore, the significance of GP2 autoantibodies in diagnosing and categorizing IBD is underscored, alongside the promising therapeutic avenues for modulating GP2 to regulate immunity and maintain mucosal balance.

Evaluation of the outcomes of the Quebec provincial suicide prevention gatekeeper training on knowledge, recognition of attitudes, perceived self-efficacy, intention to help, and helping behaviors.

Gatekeeper (GK) training is a suicide prevention strategy in which community members learn to identify individuals at risk of suicide and refer them for appropriate help. Despite its widespread use, few studies have investigated its effects, including changes in helping behaviors. To assess the impact of GK training on participants' knowledge, recognition of the influence of attitudes, perceived self-efficacy, intention to help and helping behaviors, and to identify variables associated with GK behaviors. Mixed linear effects and forward stepwise logistic regressions were used to analyze data from 159 participants receiving the Quebec Provincial GK Training program offered by five different suicide prevention centers using pretest, posttest and 6-month follow-up questionnaires. Participants' knowledge of the GK role and suicide prevention, intention to help, self-efficacy, knowledge of services, and recognition of the influence of attitudes significantly increased following training. Most changes decreased at follow-up but remained higher than at pretest. Lower levels of education and higher intention to help were significant predictors of engaging in helping behaviors in the first 6 months after receiving training. The Quebec GK training appears to be effective in preparing participants for their role but does not appear to significantly increase helping behaviors.

Therapeutic Targeting of FLT3 Gate Keeper Mutation with E2082-0047 in Traditional and a Novel Immunocompetent Murine Adoptive Transfer Model of AML

Signaling mutations such as Fms-like tyrosine kinase 3 (FLT3) internal tandem duplication (FLT3-ITD) occur in approximately 30% of AML patients which provide AML cells with a constitutively active aberrant growth advantage, resulting in inferior overall survival. Multiple phase 3 studies with different FLT3 inhibitors validated FLT3-ITD as an important therapeutic target in AML. However, resistance to FLT3-ITD inhibitors occurs via mutations in the tyrosine kinase domain (FLT3-TKD), including the gatekeeper mutation F691L for which all approved FLT3 inhibitors are ineffective. Other mechanisms of secondary FLT3 inhibitor resistance include induction of IRAK4 activity and acquisition of RAS/PTPN11/MAPK activating mutations. New FLT3 inhibitors targeting both intrinsic (FLT3 acquired mutations) and extrinsic mechanisms of FLT3 resistance (e.g. IRAK4 induction) are urgently needed. We demonstrated previously the selectivity and potency of a novel FLT3 inhibitor, E2082-0047, against FLT3-ITD and TKD mutations in vitro using a Ba/F3 cell line harboring different TKD mutations. Guided by KinomeScan data, we demonstrated simultaneous IRAK4 inhibition and verified superior in vivo efficacy compared to gilteritinib using the FLT3-ITD MOLM-13 cell line derived xenograft (CDX) murine model (Elgamal OA et al., ASH 2022). Recognizing the exceptional safety profile of E2082-0047 in pre-clinical IND enabling toxicology studies, we moved forward to test the potency of E2082-0047 versus gilteritinib in cells harboring the FLT3-ITD-F691L gatekeeper mutation using two in vivo models, MOLM-14-F691L CDX model and a novel immunocompetent murine adoptive transfer model that does not require conditioning or irradiation. This model is generated by the crossing of the CD45.2 specific Npm1 cA cre inducible model (Vassiliou et al., 2011) with the Flt3 ITD (Lee et al., 2007) model which was previously reported to harbor a F692L mutation, analogous to the human F691L mutation (Dovey et al., 2016) thus, generating the Npm1 cA;Flt3 ITD-F692L AML murine model. In the MOLM-14-F691L CDX model study, we found a consistent reduction in human CD45 cells in bone marrow and increased survival benefit in mice receiving E2082-0047 compared to gilteritinib. Further, mice in the gilteritinib group with progressing disease (> 50% human CD45+ cells in bone marrow) were switched to E2082-0047 with consequent > 5-fold reduction in bone marrow tumor burden within 8 days. These data suggest that E2082-0047 will be effective in gilteritinib resistant disease. Strikingly, in the AML adoptive transfer model, E2082-0047 resulted in a compelling long-term survival benefit. In our first study (donor #1), vehicle and gilteritinib cohorts had 29- and 32-days median survival, respectively with evident circulating CD45.2 AML. In the E2082-0047 cohort, mice remained on study with an average of <2% circulating CD45.2 cells therefore, we stopped treatment and euthanized three mice for tissue collection, leaving eight mice for disease relapse monitoring for an additional two weeks where we lost 3/8 mice due to early removal criteria (ERC) and 5/8 were euthanized at end of study day 100, with an average of <10% circulating CD45.2 cells. This initial study showed the capacity for E2082-0047 to induce long-term survival in a model with a clinically relevant TKD gatekeeper mutation. We repeated our study with a more aggressive donor (donor #2) where vehicle and gilteritinib arms had a median survival of 22 days. While mice receiving E2082-0047 remain on study (week 8 of an ongoing study), with an average of <5% peripheral blood CD45.2 level (Figure 1). Collectively, our previous and follow-up studies herein demonstrate the superiority of E2082-0047 over gilteritinib against FLT3-ITD and with TKD mutations including the gatekeeper F691L variant. We demonstrate significantly improved survival after E2082-0047 treatment in an immune deficient in vivo model of FLT3-ITD/TKD-F691L MOLM-14 AML and a durable long-term survival in a large subset of mice using two donors in an immune competent adoptive transfer model of NPM1+FLT3-ITD/TKD-F692L AML. Given the curative potential of E2082-0047 in the immune competent model, ongoing efforts are focused on investigating the host immune response to AML. Given the safety and efficacy studies performed with E2082-0047 to date, a healthy volunteer study to best estimate dose for trials in AML is ongoing.

Design and Adoption of a FHIR IG to Support a Telemonitoring Environment in Gatekeeper Project.

Telemedicine can provide benefits in patient affected by chronic diseases or elderly citizens as part of standard routine care supported by digital health. The GATEKEEPER (GK) Project was financed to create a vendor independent platform to be adopted in medical practice and to demonstrate its effect, benefit value, and scalability in 8 connected medical use cases with some independent pilots. This paper, after a description of the GK platform architecture, is focused on the creation of a FHIR (Fast Healthcare Interoperability Resource) IG (Implementation Guide) and its adoption in specific use cases. The final aim is to combine conventional data, collected in the hospital, with unconventional data, coming from wearable devices, to exploit artificial intelligence (AI) models designed to evaluate the effectiveness of a new parsimonious risk prediction model for Type 2 diabetes (T2D).

Structural and functional insights of a cold-adaptive β-glucosidase with very high glucose tolerance from Microbacterium sp. CIAB417

A gene glu1 (WP_243232135.1) coding for β-glucosidase from the genome of Microbacterium sp. CIAB417 was characterized for its cold adaptive nature and tolerance to high levels of glucose and ethanol. The phylogenetic analysis suggested the close association of glu1 with a similar gene from a mesophilic bacterium Microbacterium indicum. The purified recombinant GLU1 displayed its optimal activity and stability at pH 5 and temperature 30ᴼC. Additionally, the presence of L3 loop in GLU1 suggested its cold adaptive nature. The glucose tolerant Gate keeper residues (Leu 174 & Trp 169) with a distance of ∼ 6.953 Å between them was also predicted in GLU1. The GLU1 enzyme showed ≥ 95% and ≥ 40% relative activity in the presence of 5 M glucose and 20% ethanol. The Vmax, Km, and Kcat values of GLU1 for cellobiose substrate were observed to be 45.22 U/mg, 3.5 mM, and 41.0157 s-1, respectively. The GLU1 was found to be highly efficient in hydrolysis of celloologosaccharides (C2-C5), lactose and safranal picrocrocin into glucose. Hence, cold adaptive GLU1 with very high glucose and ethanol tolerance could be very useful in bio-refinery, dairy, and flavor industries.

Criminal Responsibility in Train Accidents at Level Crossings in Indonesia

The accident involving the 112 Brantas train bound for Pasar Senen-Blitar with a tronton truck on Jalan Madukoro, Semarang City, Central Java, left grief. Police officials have not been able to confirm who is guilty and can be held criminally responsible for the accident. This study aims to determine criminal responsibility in a train accident at a level crossing, the research method used is normative legal research which is prescriptive by examining library based materials and secondary legal materials in the form of legal rules, legal principles, and legal doctrines to produce arguments, theories or new concepts as prescriptions in resolving the problems at hand. The results of the study show that, First, those who can be held criminally liable for the accident are legal subjects who intentionally or negligently interfere with train travel and are responsible for opening the road at level crossings. Engineers and crossing gatekeepers cannot be held criminally liable because the task of the Train Engineer and Crossing Gate Keeper is to ensure that trains can run on the rails in accordance with the allotment of railroads as stipulated in the railway law.

A Study on the Efficiency of Disaster Information Communication Based on the Gate Keeper Theory

In recent years, major disasters have occurred frequently, with sudden floods, rampant epidemics, and typhoons... Faced with sudden disasters, the affected population is often caught unprepared. Under the background of “Internet +”, the speed of information circulation has improved, but the news that the material needs of the disaster victims are not effectively met still often appears in the public’s view. In order to improve the efficiency of disaster information dissemination and enable the efficient distribution of related materials, this paper discusses this issue based on the gatekeeper theory of communication science. With a view to tracing the source and clarifying the reasons for the low efficiency and poor effect of disaster mutual aid information dissemination under the background of “Internet +”, this paper proposes solutions to the problems in combination with the gatekeeper theory of communication, to promote the rational distribution and utilization of resources, meet the material needs of disaster victims, and ensure the basic life of the people.

Analysis of the ERK Pathway Cysteinome for Targeted Covalent Inhibition of RAF and MEK Kinases

The ERK pathway is one of the most important signaling cascades involved in tumorigenesis. So far, eight noncovalent inhibitors of RAF and MEK kinases in the ERK pathway have been approved by the FDA for the treatment of cancers; however, their efficacies are limited due to various resistance mechanisms. There is an urgent need to develop novel targeted covalent inhibitors. Here we report a systematic study of the covalent ligandabilities of the ERK pathway kinases (ARAF, BRAF, CRAF, KSR1, KSR2, MEK1, MEK2, ERK1, and ERK2) using constant pH molecular dynamics titration and pocket analysis. Our data revealed that the hinge GK (gate keeper)+3 cysteine in RAF family kinases (ARAF, BRAF, CRAF, KSR1, and KSR2) and the back loop cysteine in MEK1 and MEK2 are reactive and ligandable. Structure analysis suggests that the type II inhibitors belvarafenib and GW5074 may be used as scaffolds for designing pan-RAF or CRAF-selective covalent inhibitors directed at the GK+3 cysteine, while the type III inhibitor cobimetinib may be modified to label the back loop cysteine in MEK1/2. The reactivities and ligandabilities of the remote cysteine in MEK1/2 and the DFG-1 cysteine in MEK1/2 and ERK1/2 are also discussed. Our work provides a starting point for medicinal chemists to design novel covalent inhibitors of the ERK pathway kinases. The computational protocol is general and can be applied to the systematic evaluation of covalent ligandabilities of the human cysteinome.

Abstract 3474: Identification of the selective CBL-b inhibitors which effectively prevent T cells from exhaustion and demonstrate synergistic anti-tumor activity in combination with an anti-PD1 antibody

Abstract The E3 ubiquitin ligase Casitas B-lineage lymphoma b (Cbl-b) is a member of the highly conserved family of Cbl (casitas b-lineage lymphoma) proteins. CBL-b is widely expressed in immune cells and plays as a “gate keeper” to negatively regulate the activation of immune cells. Engagement of TCR/BCR triggers the Cbl-b-mediated ubiquitination and degradation of the key components in TCR axis, such as Vav1 and PLCγ, and consequently attenuates TCR/BCR signal axis which finally results in T/B cells exhaustion. Besides, Cbl-b was reported to negatively regulate the activation of dendritic cells, NK, NKT, and mast cells. Cbl-b KO mice show resistance to tumor transplantation, and the anti-tumor activities depend on the T and NK cells. According to the above observation, Cbl-b emerges as a novel intracellular immune checkpoint target to prevent immune cells from exhaustion and improve the antitumor immunotherapy. Here we identified a series of highly potent Cbl-b inhibitors. The inhibition of Cbl-b protein was evaluated by an in vitro CBL-b/E2-ubiquitin assay. Results showed that our candidates strongly inhibited Cbl-b with IC50s at single-digital nanomolar level, and meanwhile, these compounds displayed over than 10-fold selectivity against c-CBL/E2-ubiquitin, a member of the Cbl family, which was reported to synergistically enhance toxicity caused by Cbl-b. Consistent with the Cbl-b inhibition, these compounds further enhanced activation of Jurkat T cells as well as human PBMCs with dramatical increase of IL-2 or IFNɣ secretion in presence of anti-CD3/CD28, and without any significant cytotoxicity even up to 10 µM concertation. Furthermore, these compounds exhibited the restoring function in exhausted T cells in presence with PEG2 or adenosine treatment. Favorable ADME and bioavailability supported the oral administration. In naïve mice, our inhibitors dose-dependently promoted the cytokine IL-2 and IFNɣ production in serum and spleens following the anti-CD3 challenge. In a syngeneic model, CT26, oral administration of our Cbl-b inhibitor effectively inhibited tumor growth, while completely inhibited tumor growth in combination with an anti-PD1 antibody. Further study demonstrated that the increased lymphocytes infiltration and cytokines release in tumor tissues were observed either in Cbl-b mono or in anti-PD1 combo groups, which correlated well with anti-tumor efficacy. In conclusion, our selective Cbl-b inhibitors demonstrated robust immune activation and anti-tumor activity in vitro and in vivo and showed synergistic activities with immune checkpoint inhibitors, such as anti-PD(L)1. Currently, one of our leading molecules is in IND enabling studies. Citation Format: Feng Zhou, Zhengtao Li, Yajing Liu, Guimei Yang, Wansun Mai, Wenqing Yang, Liting Xue, Ping Chen, Renhong Tang. Identification of the selective CBL-b inhibitors which effectively prevent T cells from exhaustion and demonstrate synergistic anti-tumor activity in combination with an anti-PD1 antibody [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3474.

Comparison of genetic diversity and phylogenetic structure of BRCA1 gene of some domestic and wild sheep breeds in different countries

BRCA1 gene plays an important role in DNA damage repair, cell cycle, and transcription process regulation; hence it’s called gate keeper. The current research aims to perform bioinformatics analyzes of the BRCA1 gene of different breeds of domestic and wild sheep from 49 breeds in 14 countries using the NCBI genome database. The desired sequences were aligned using MEGA11 software and a phylogenetic tree was drawn by Neighbor-Joining method. The number of mutations, nucleotide diversity, and haploid diversity were also analyzed using Dnaspv5 software. The analyses showed 296 polymorphisms, which led to the creation of 45 different haplotypes with a haplotype diversity of 0.035. Nucleotide diversity and average nucleotide differences among breeds were estimated as 0.259 and 0.052, respectively. The average genetic distance within the population of countries was calculated as 0.052. The amount of sequence conservation in this research was 0.313 on average, which indicates the high polymorphism of this gene and the emergence of new proteins. Tajima’s D value in Tajima’s neutrality test was −2.421, which was significant (p < 0.05). One of the reasons for the high genetic diversity in Iran’s wild sheep population is the existence of forests and open environments, which prevent genetic drift and reduce inbreeding.

A novel smart stealth sorafenib delivery system based on the magnetic imprinting material modified by polyethylene glycol

AbstractIn order to improve the solubility and the long‐cycle performance of hydrophobic drug sorafenib (SOR), and to obtain an ideal smart drug delivery system for the effective treatment of liver cancer, S‐SMIP, a pH/GSH double‐sensitive sorafenib stealth imprinting material, was successfully prepared by UV‐initiated surface imprinted polymerization on the basis of polyethylene glycol 2000 (PEG2000) acted as “gate keeper” modified magnetic substrate. The results showed that S‐SMIP had a good adsorption capacity of 260.0 mg g−1 and an excellent selectivity factor α 2.33. S‐SMIP's kinetic adsorption data accorded well with the pseudo‐second‐order model and its mechanism was controlled mainly by chemical adsorption. The drug release experiments in vitro indicated S‐SMIP could not only greatly improve the solubility of SOR, but also had the pH/GSH dual‐sensitive response performance. Compared with the non‐stealth imprinted material SMIP, S‐SMIP still had a better long‐cycle effect on the release of SOR, increasing to160 h from 130 h. So, the target material S‐SMIP could greatly prolong the half‐life of SOR, which provided a good idea for the development of a smart drug delivery system.

Accumulation of lankamycin derivative with a branched-chain sugar from a blocked mutant of chalcose biosynthesis in Streptomyces rochei 7434AN4

Lankamycin, a macrolide antibiotic produced by Streptomyces rochei 7434AN4, exhibits a moderate antimicrobial activity and acts as a synergistic pair with carbocyclic antibiotic lankacidin C by binding to the ribosome exit tunnel. Its biosynthetic gene (lkm) cluster (orf24-orf53) is located on the largest plasmid pSLA2-L (210,614 bp). Our group possesses a variety of lankamycin derivatives and macrolide-modification enzymes including P450 enzymes and glycosyltransferases, which may lead to expand the chemical library of bioactive macrolides. Here we constructed a mutant of a 3-ketoreductase gene lkmCVI (orf42) involved in d-chalcose biosynthesis, and its metabolite was isolated and structure-elucidated. Accumulation of novel lankamycin derivative harboring a branched-chain deoxysugar, 5-O-(4′,6′-dideoxy-3′-C-acetyl-d-ribo-hexopyranosyl)-3-O-(4″-O-acetyl-l-arcanosyl)-lankanolide, indicated that LkmCVI acts as a gate keeper enzyme for d-chalcose synthesis in lankamycin biosynthesis.

Iron level changes in the brain with neurodegenerative disease

Nanosized magnetite inside ferritin may control the progression of neurodegenerative disease (ND) by creating an electronic noise in the neural cells. We discovered that brains with ND have a unique electron paramagnetic resonance (EPR) spectrum. Our measurements showed that the collapse of the broad ferritin maximum contained in the EPR spectra possibly relates to the onset and progression of the ND. Ferritin malfunction triggers the perturbation of iron concentration that either increases or decreases over the normal levels in brain without ND. This supports a conjecture that accumulated iron results in an increased volume of magnetite crystals, whose fluctuated magnetic moments may interfere with the normal function of neural synapses and contribute to the neurodegenerative disease. The mechanism of the iron mobility relates to iron canals in the neural cell's membrane by which the iron enters and leaves the neural cells. This gate keeper malfunction may relate to a speculation that this is due to the appearance of 2Fe-2S in EPR spectra of brains with ND. Statement of significanceIn this manuscript we describe feedback between electronic structure of atoms in the brain, easiness of becoming magnetized in a magnetic field and the ability of the brain to hold the magnetic field on its own in cases of neurodegenerative-diseased and healthy brain. This contribution is novel and significant for a number of reasons, as follows: We revealed that diseased brains have a distinct electronic structure from healthy brains. We identified the easiness of brain samples to become magnetized in a magnetic field and the brains’ ability to hold the magnetic field on its own in cases of neurodegenerative-diseased and healthy brains. This paper addresses a new hypothesis, and we consider that it will generate broad that may be of broad interdisciplinary interest and generate further debate.

COMMITMENT-BASED CAPITATION EVALUATION IN BENGKULU PROVINCE: A REALIST EVALUATION

Primary Health Care (FKTP) as the gate keeper of health service providers must have a strategy in improving service quality and patient satisfaction. This study aims to evaluate the implementation of regulations regarding commitment-based capitation policies and determine the mechanism of change and contextual factors that affect the implementation of regulations. The evaluation was carried out using a realist evaluation approach and analyzed in terms of the configuration of forms, mechanisms, and results. The research was carried out in Bengkulu Province in 2018-2019 with the research subjects consisting of the health office, the head of the Puskesmas and the JKN monev team, Private Practice Doctors, Pratama Clinics. The sample selection used purposive sampling and data collection was carried out through in-depth interviews. Commitment-Based Capitation (KBK) is running in Bengkulu province. The KBK program shows that in general the non-specialist referral indicator target in Bengkulu province is achieved, namely 5%, but there are also city districts that have not been achieved, this is achieved because the Puskesmas has doctors in Bengkulu province, adequate doctor competence, adequate facilities and infrastructure, staff receive capitation funds incentives, there is a JKN Monev team that is always active in conducting socialization, monitoring and evaluation in all Puskesmas, especially Puskesmas that do not achieve non-specialist referral indicators. This study provides an overview of the context, mechanism and outcome of health service quality policy regulations in the JKN era, especially the Commitment-Based Capitation program.&#x0D; &#x0D;

Discovery of 2-Amino-7-sulfonyl-7H-pyrrolo[2,3-d]pyrimidine Derivatives as Potent Reversible FGFR Inhibitors with Gatekeeper Mutation Tolerance: Design, Synthesis, and Biological Evaluation.

Fibroblast growth factor receptors (FGFRs) play key roles in promoting cancer cell proliferation, differentiation, and migration. However, acquired resistance to FGFR inhibitors has become an emerging challenge in long-term cancer therapies, especially for hepatocellular carcinoma (HCC). Gatekeeper (GK) mutations are the main mechanism of resistance. Herein, we describe the discovery of a series of reversible FGFR inhibitors, particularly for GK mutations with the 2-amino-7-sulfonyl-7H-pyrrolo[2,3-d]pyrimidine scaffold. Rational design, optimization, and pharmacokinetic screening provided representative compound 19 with potent FGFR inhibition in vitro, high bioavailability, and an acceptable half-life. GK mutation tolerance was supported by assays against FGFR4V550L and Ba/F3-TEL-FGFR4V550L cells. Moreover, compound 19 exhibited potent antitumor potency in HUH7 xenograft mouse models with no obvious toxicity observed. Compound 19 was identified as a potential candidate for overcoming GK mutations for HCC treatment.

Dual responsive block copolymer coated hollow mesoporous silica nanoparticles for glucose-mediated transcutaneous drug delivery

A self-regulated anti-diabetic drug release device mimicking pancreatic cells is highly desirable for the therapy of diabetes. Herein, a glucose-mediated dual-responsive drug delivery system, which combines pH- and H 2 O 2 -responsive block copolymer grafted hollow mesoporous silica nanoparticles (HMSNs) with microneedle (MN) array patch, has been developed to achieve self-regulated administration. The poly[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl acrylate]- b -poly[2-(dimethylamino)ethyl methacrylate] (PPBEM- b -PDM) polymer serves as gate keeper to prevent drug release from the cavity of HMSNs at normoglycemic level. In contrast, the drug release rate is significantly enhanced upon H 2 O 2 and pH stimuli due to the chemical change of H 2 O 2 sensitive PPBEM block and acid responsive PDM block. Therefore, incorporation of anti-diabetic drug and glucose oxidase (GOx, which can oxidize glucose to gluconic acid and in-situ produce H 2 O 2 ) into stimulus polymer coated HMSNs results in a glucose-mediated MN device after depositing the drug-loaded nanoparticles into MN array patch. Both in vitro and in vivo results show this MN device presents a glucose mediated self-regulated drug release characteristic, which possesses a rapid drug release at hyperglycemic level but retarded drug release at normoglycemic level. The result indicates that the fabricated smart drug delivery system is a good candidate for the therapy of diabetes.