Non-specific Control Research Articles

A biomimetic enzyme-linked immunosorbent assay (BELISA) for the analysis of gonadorelin by using molecularly imprinted polymer-coated microplates

An original biomimetic enzyme-linked immunoassay (BELISA) to target the small peptide hormone gonadorelin is presented. This peptide has been recently listed among the substances banned in sports by the World Antidoping Agency (WADA) since its misuse by male athletes triggers testosterone increase. Hence, in response to this emerging issue in anti-doping controls, we proposed BELISA which involves the growth of a polynorepinephrine (PNE)–based molecularly imprinted polymer (MIP) directly on microwells. PNE, a polydopamine (PDA) analog, has recently displayed impressive performances when it was exploited for MIP preparation, giving even better results than PDA. Gonadorelin quantification was accomplished via a colorimetric indirect competitive bioassay involving the competition between biotinylated gonadorelin linked to the signal reporter and the unlabeled analyte. These compete for the same MIP binding sites resulting in an inverse correlation between gonadorelin concentration and the output color signal (λ = 450 nm). A detection limit of 277 pmol L−1 was achieved with very good reproducibility in standard solutions (avCV% = 4.07%) and in urine samples (avCV% = 5.24%). The selectivity of the assay resulted adequate for biological specimens and non-specific control peptides. In addition, the analytical figures of merit were successfully validated by mass spectrometry, the reference anti-doping benchtop platform for the analyte. BELISA was aimed to open real perspectives for PNE-based MIPs as alternatives to antibodies, especially when the target analyte is a poorly or non-immunogenic small molecule, such as gonadorelin.Graphical abstractBiomimetic enzyme-linked immunosorbent assay (BELISA)

An Ultrasensitive Silicon-Based Electrolyte-Gated Transistor for the Detection of Peanut Allergens.

The highly sensitive detection of peanut allergens (PAs) using silicon-based electrolyte-gated transistors (Si-EGTs) was demonstrated. The Si-EGT was made using a top-down technique. The fabricated Si-EGT showed excellent intrinsic electrical characteristics, including a low threshold voltage of 0.7 V, low subthreshold swing of <70 mV/dec, and low gate leakage of <10 pA. Surface functionalization and immobilization of antibodies were performed for the selective detection of PAs. The voltage-related sensitivity (SV) showed a constant behavior from the subthreshold regime to the linear regime. The current-related sensitivity (SI) was high in the subthreshold regime and then significantly decreased as the drain current increased. The limit of detection (LOD) was calculated to be as low as 25 pg/mL based on SI characteristics, which is the lowest value reported to date in the literature for various sensor methodologies. The Si-EGT showed selective detection of PA through a non-specific control test. These results confirm that Si-EGT is a high-sensitivity and low-power biosensor for PA detection.

Creation of a model for studying the antiviral effect of small interfering RNAs in vitro

Influenza is a widespread respiratory infection, accompanied by damage to the lower respiratory tract of a person, and can lead to severe complications, up to a fatal outcome. Currently existing therapeutic agents and vaccines do not provide fully effective protection against influenza viruses. The development and creation of drugs based on the mechanism of RNA interference in the context of this problem is a promising direction. The aim of this study is to select and experimentally substantiate cellular target genes for miRNAs whose knockdown suppresses viral reproduction. A549 cells (human lung adenocarcinoma) were transfected with small interfering RNAs. After 4 hours, the transfected cells were infected with the influenza virus pri MOI = 0.1; 0.01 and 0.001. The virus-containing liquid was sampled within three days from the moment of transfection and the intensity of the dynamics of viral reproduction was assessed by the CPD titration method. The use of all small interfering RNAs at MOI = 0.1 resulted in a significantly significant decrease in the viral titer relative to non-specific control. Similar results were obtained at MOI = 0.01 and 0.001. The most effective siRNA ISSINUP98, since when using it, at MOI = 0.1, the viral titer values decreased by 1.7 lg TCD50/ml and by 3 lg TCD50/ml at MOI = 0.01 on the third day. The results showed that miRNAs directed to human cellular genes FLT4, Nup98 and Nup205, whose derivatives play an important role in the life cycle of the influenza virus, effectively reduce its reproduction in vitro. Thus, the studied genes and their products are potential targets for the development of anti-influenza drugs.

Mobile phone-based interventions for mental health: A systematic meta-review of 14 meta-analyses of randomized controlled trials.

Mobile phone-based interventions have been proposed as a means for reducing the burden of disease associated with mental illness. While numerous randomized controlled trials and meta-analyses have investigated this possibility, evidence remains unclear. We conducted a systematic meta-review of meta-analyses examining mobile phone-based interventions tested in randomized controlled trials. We synthesized results from 14 meta-analyses representing 145 randomized controlled trials and 47,940 participants. We identified 34 effect sizes representing unique pairings of participants, intervention, comparisons, and outcome (PICO) and graded the strength of the evidence as using umbrella review methodology. We failed to find convincing evidence of efficacy (i.e., n > 1000, p < 10−6, I2 < 50%, absence of publication bias); publication bias was rarely assessed for the representative effect sizes. Eight effect sizes provided highly suggestive evidence (i.e., n > 1000, p < 10−6), including smartphone interventions outperforming inactive controls on measures of psychological symptoms and quality of life (ds = 0.32 to 0.47) and text message-based interventions outperforming non-specific controls and active controls for smoking cessation (ds = 0.31 and 0.19, respectively). The magnitude of effects and strength of evidence tended to diminish as comparison conditions became more rigorous (i.e., inactive to active, non-specific to specific). Four effect sizes provided suggestive evidence, 14 effect sizes provided weak evidence, and eight effect sizes were non-significant. Despite substantial heterogeneity, no moderators were identified. Adverse effects were not reported. Taken together, results support the potential of mobile phone-based interventions and highlight key directions to guide providers, policy makers, clinical trialists, and meta-analysts working in this area.

Creation of a model for studying the antiviral effect of small interfering RNAs in vitro

Influenza is a widespread respiratory infection, accompanied by damage to the lower respiratory tract of a person, and can lead to severe complications, up to a fatal outcome. Currently existing therapeutic agents and vaccines do not provide fully effective protection against influenza viruses. The development and creation of drugs based on the mechanism of RNA interference in the context of this problem is a promising direction. The aim of this study is to select and experimentally substantiate cellular target genes for miRNAs whose knockdown suppresses viral reproduction. A549 cells (human lung adenocarcinoma) were transfected with small interfering RNAs. After 4 hours, the transfected cells were infected with the influenza virus pri MOI = 0.1; 0.01 and 0.001. The virus-containing liquid was sampled within three days from the moment of transfection and the intensity of the dynamics of viral reproduction was assessed by the CPD titration method. The use of all small interfering RNAs at MOI = 0.1 resulted in a significantly significant decrease in the viral titer relative to non-specific control. Similar results were obtained at MOI = 0.01 and 0.001. The most effective siRNA ISSINUP98, since when using it, at MOI = 0.1, the viral titer values decreased by 1.7 lg TCD50/ml and by 3 lg TCD50/ml at MOI = 0.01 on the third day. The results showed that miRNAs directed to human cellular genes FLT4, Nup98 and Nup205, whose derivatives play an important role in the life cycle of the influenza virus, effectively reduce its reproduction in vitro. Thus, the studied genes and their products are potential targets for the development of anti-influenza drugs.

Design of primers in the molecular detection of Feline Panleukopenia Virus

Feline Panleukopenia is a disease characterized by a reduction in the number of circulating leukocytes and enteritis with degeneration of the intestinal villi. The etiologic agent, called Feline Panleukopenia virus (FPV), belongs to the Parvoviridae family, is highly contagious and has high mortality and morbidity. Although vaccination of healthy cats is the most effective way to prevent the disease, once the symptoms appear, the treatment is supportive, presenting high mortality in the first days of the disease. FPV positive cats should be hospitalized and isolated for at least 2 weeks to avoid viral transmission. Early detection is usually done with the enzyme-linked immunoadsorption assay (ELISA) test that detects viral antigens in stool samples. As a complementary diagnostic method, in this work it was proposed to implement the Polymerase Chain Reaction (PCR) aimed at the diagnosis of FPV initially in positive samples from two feline vaccines and one canine vaccine. As an approximation to real samples, the commercial vaccines were mixed with feces and blood from a healthy cat. The results showed that the In Silico design was successful strategy based on the VP2 gene sequences of FPV available in GenBank® in conjunction with the sharp visualization of positive controls of the expected size and without amplification. nonspecific or negative controls. The fragments obtained has a nucleotide identity percentage greater than 97% with respect to the information available in Genbank® and corroborates the detection of a VP2 fragment. Thus, the future option of applying the same protocol in a diagnostic way is opened, using samples obtained from suspected patients who are infected with FPV.

The oral and gut microbiota: beyond a short communication

Introduction. The current treatment and prevention of oral disorders, dental caries, periodontal and gum diseases, follow a very non-specific control of plaque as the main causative factor. The main therapeutically approach is carried out on the sole perspective to keep the levels of oral bacteria in an acceptable range compatible with one-way vision of oral-mouth health, as something completely separated from a systemic microbial homeostasis (dysbiosis) concomitant present in the gut. A sealed compartmental view which sees separate and incommunicable responses to a specific condition without considering the presence of interacting confounding factors can negatively influence the diagnosis a diseases and of course its progression. A general non-specific antimicrobial with more general antiplaque therapy based mainly on oral care products together with surgery interventions represent at the moment the only mechanical responses in treating oral diseases. Material and method. The present paper is a narrative review concening interractions between oral and gut microbiota, with a focus on the interdisciplinary approach in antimicrobial treatment. Pubmed, Cochrane Library database were used for searching engines. Key words used were as follows: “inflammatory bowel syndrome (IBS)”, “ulcerative colitis”, “oral dysbiosis”, “gut dysbiosis”, “probiotics”, “periodontitis”. Results and discussions. Literature research showed that there are few issues to be discussed the ever increasing resistance to antibiotics, the high consumption of industrial food and sugars and their negatively effect on gut and oral microbiota. There is a need to highlight and develop a novel philosophical approach in the treatments for oral diseases that will necessarily involve non-conventional antimicrobial solutions. Such approaches should preferably reduce the consumption of both intestinal and oral microbiota, that are intimately connected and host approximately well over 1000 different species of bacteria at 108–109 bacteria per mL of mucous and saliva. Preventive approaches based upon the restoration of the microbial ecological balance, rather than elimination of the disease associated species, have been proposed. Conclusions. Having both oral-gut microbiota screened is an essential moment that influence the healthy immune modulatory and regenerative capacity of the body and, the new proposed formula integrates a wider screen on the patients where oral condition is strictly evaluated together with gut screen; therefore any proposed treatment will be inevitably sustained by the use of prebiotics and probiotics to promote health-associated bacterial growth. Keywords: inflammatory bowel syndrome (IBS), ulcerative colitis, oral dysbiosis, gut dysbiosis, probiotics, periodontitis,

Computational Simulation of HIV Protease Inhibitors to the Main Protease (Mpro) of SARS-CoV-2: Implications for COVID-19 Drugs Design.

SARS-CoV-2 is highly homologous to SARS-CoV. To date, the main protease (Mpro) of SARS-CoV-2 is regarded as an important drug target for the treatment of Coronavirus Disease 2019 (COVID-19). Some experiments confirmed that several HIV protease inhibitors present the inhibitory effects on the replication of SARS-CoV-2 by inhibiting Mpro. However, the mechanism of action has still not been studied very clearly. In this work, the interaction mechanism of four HIV protease inhibitors Darunavir (DRV), Lopinavir (LPV), Nelfinavir (NFV), and Ritonavire (RTV) targeting SARS-CoV-2 Mpro was explored by applying docking, molecular dynamics (MD) simulations, and MM–GBSA methods using the broad-spectrum antiviral drug Ribavirin (RBV) as the negative and nonspecific control. Our results revealed that LPV, RTV, and NFV have higher binding affinities with Mpro, and they all interact with catalytic residues His41 and the other two key amino acids Met49 and Met165. Pharmacophore model analysis further revealed that the aromatic ring, hydrogen bond donor, and hydrophobic group are the essential infrastructure of Mpro inhibitors. Overall, this study applied computational simulation methods to study the interaction mechanism of HIV-1 protease inhibitors with SARS-CoV-2 Mpro, and the findings provide useful insights for the development of novel anti-SARS-CoV-2 agents for the treatment of COVID-19.

Present-day tendencies in accompanying therapy in cancer treatment: from theory of adaptational reactions to bio-information technologies translation

At present, to develop new, scientifically justified, technologiesof an accompanying therapy to improve the oncological situationis a challenge to health care. A therapy to accompany aspecific cancer treatment in case of a malignant process in anorganism is dictated by the necessity to raise the nonspecific resistanceof the organism in cancer patients. In this case, originalmethodological approaches to initiate the mechanisms of thenonspecific resistance have been developed on the basis of thetheory of the adaptational reactions. Within the framework ofthe above theory, some concepts of relationship between thespecific and nonspecific anti-tumor processes have been extensivelydeveloped that may be successfully applied to solvingtasks of the National Health Care Program.Aim. The aim hereof is to introduce the theoretical basics ofthe nonspecific adaptation regulation and control by a tumor-bearing organism into practice for an integrated treatmentof malignant tumors with the use of the activation therapy technologies.Materials and methods. Our research work has involvedclinical data on 1310 patients, who have been diagnosedwith cancer of different localization: breast cancer (n = 170),cervical cancer (n = 60), lung cancer (n = 760), bladder cancer(n = 120), brain gliomas of high grade of malignancy andmetastatic damage (n = 170) and extended colorectal cancer(n = 30) and who have completed their treatment at the NationalMedical Research Centre of Oncology of the Ministry ofHealth of the Russian Federation.

EPCO-05. GENOME-WIDE ANALYSIS OF TEAD1 OCCUPANCY IN BIOLOGICALLY DISTINCT GLIOBLASTOMA SAMPLES

Abstract The diffusely infiltrative nature of glioblastoma (GBM) cells is a major contributor to the disease’s aggressive behavior, including its rapid progression and therapeutic resistance. Moreover, current treatment options do not target the invasive nature of GBM. Recent chromatin accessibility studies prioritized enrichment of the TEAD1 transcription factor motif in glioblastoma stem cell biology and subsequent knockout and overexpression studies confirmed a critical role for TEAD1 in GBM migration, in vitro and in vivo. However, the downstream mechanisms through which TEAD1 regulates GBM cell migration remain poorly understood. In this study, we performed chromatin immunoprecipitation (ChIP-seq) using TEAD1-specific antibody and IgG as non-specific binding control, to characterize TEAD1 occupancy across GBM samples with unique genomic alterations. ChIP-seq peaks were called using MACS2, filtered for duplicates and blacklisted regions, and normalized per sample to their respective genomic input. Initial functional enrichment analyses were performed on three GBM samples with the highest number of TEAD1 occupancy peaks using CistromeGO, which ranked genes based on their TEAD1-specific regulatory potential (RP) score, as a function of peak number and distance from the transcription start site. Analyses of the top 1000 genes with highest TEAD1 RP scores identified 132 common targets across all samples, including known TEAD target genes ETV1 and Cyr61, which related to angiogenesis, cadherin and integrin signaling, cell adhesion, and chromatin regulation gene ontology terms, among others. Interestingly, KEGG pathway analysis also revealed Hippo pathway enrichment across all GBM samples, suggesting a possible TEAD1 regulatory feedback loop in GBM. Analysis of TEAD1 ChIP-seq peaks in non-GBM negative control tissue did not show functional enrichment for any of the terms seen in the GBM samples. Ongoing analyses are focused on characterizing TEAD1 occupancy at active cis-regulatory regions using parallel H3K27ac ChIP-seq data, in order to prioritize the most salient TEAD1-regulatory targets in GBM.

Utility of TMPRSS4 as a Prognostic Biomarker and Potential Therapeutic Target in Patients with Gastric Cancer

BackgroundTransmembrane serine protease 4 (TMPRSS4) belongs to the family of type II transmembrane serine proteases that are known to be upregulated in many malignant tumors. However, there is a paucity of studies documenting the clinical impact and biological effects of TMPRSS4 on gastric cancer (GC) patients who underwent surgery. MethodsTissues samples were obtained from 105 patients with GC who underwent gastrectomy followed by adjuvant chemotherapy, excluding those at stage I. The expression of TMPRSS4 was examined through immunohistochemical analysis. The association between TMPRSS4 expression and clinico-pathological features as well as prognosis was assessed. Moreover, the effects of TMPRSS4 expression on cell migration and sensitivity to 5-FU were investigated. ResultsThe expression rate of TMPRSS4 was 56.3% (59/105) in GC cases. The expression of TMPRSS4 was positively correlated with the depth of tumor (T) and venous (V) invasion. The 5-year overall survival (OS) and recurrence-free survival (RFS) rates of the TMPRSS4-positive group was significantly lower than that of the TMPRSS4-negative group (p=0.0001 and p=0.005, respectively). Especially, there was significant differences in OS and RFS of patients with stage III cancer between the two groups (p=0.0064 and 0.012, respectively). Multivariate analysis demonstrated that TMPRSS4 expression and the stage of cancer were crucial prognostic factors for RFS. TMPRSS4-silenced GC cells exhibited increased sensitivity to 5-FU when compared with the non-specific control siRNA-transfected cells. ConclusionTMPRSS4 can be considered as a potential prognostic biomarker, especially for stage III, and a promising therapeutic target for GC.

Management of the Oral Health of Children During the COVID-19 Pandemic in Poland.

Managing the oral health of children during the time of a health emergency linked to the current COVID-19 pandemic presents specific problems. A high number of non-specific effective infection control protocols are available in dental settings. It is of fundamental importance to implement specific protocols relating to those clinical situations that normally do not represent an emergency but which now fall into that category. The aim of this study was the comparison of data obtained from the Regional National Health Fund (NFZ) relating to the number and the type of procedures in the oral health management of children aged 0–18 years from the Wielkopolska region, with the months of March and April of 2019 being compared with those of the, respective, pandemic period of 2020. The results showed statistical differences in the number of performed procedures when comparing 2019 and 2020; especially in April (n = 53,077 in 2019 but only n = 2,287 in 2020), when lockdown restrictions reached their highest level and when only 30% of the dental clinics for children were open for patients in the Wielkopolska region of Poland. Regarding surgical cases, there were no differences in percentage frequency between April 2019 and 2020 in terms of extractions. However, an increase was observed in abscess incisions (3.5–17.8%) and surgical dressings (1.5–10.07%). There was a decrease in the total number of performed conservative dentistry procedures in April 2020, but temporary fillings in primary and permanent teeth showed a prominent increase: from 6.4% in 2019 to 19.3% in 2020; and 5.8–11.4%, respectively. Pulp treatment and mucosal lesions therapy fall into the dental emergency category during this COVID-19 pandemic. These cases have shown an increase from 3.2% in 2019 to 12.8% in 2020 for pulp treatment, and from 2.3 to 4.3% for the treatment of oral mucosal lesions. As suspected, after the lockdown was implemented, the number of pediatric dental cases were low. Moreover, the analysis revealed differences in the profile of clinical situations that represented the emergency cases and the pandemic treatment protocols. Future implications suggest that dental prophylactic procedures be included in pandemic protocols with even dental services being limited to a form of urgent treatment. New approaches and treatment models should be implemented in the control of the infectious spread of the disease in the management of the oral health of children in this pandemic period.

Expert consensus on the important chronic non-specific neck pain motor control and segmental exercise and dosage variables: An international e-Delphi study.

Chronic non-specific neck pain is highly prevalent, resulting in significant disability. Despite exercise being a mainstay treatment, guidance on optimal exercise and dosage variables is lacking. Combining submaximal effort deep cervical muscles exercise (motor control) and superficial cervical muscles exercise (segmental) reduces chronic non-specific neck pain, but evaluation of optimal exercise and dosage variables is prevented by clinical heterogeneity. To gain consensus on important motor control and segmental exercise and dosage variables for chronic non-specific neck pain. An international 3-round e-Delphi study, was conducted with experts in neck pain management (academic and clinical). In round 1, exercise and dosage variables were obtained from expert opinion and clinical trial data, then analysed thematically (two independent researchers) to develop themes and statements. In rounds 2 and 3, participants rated their agreement with statements (1-5 Likert scale). Statement consensus was evaluated using progressively increased a priori criteria using descriptive statistics. Thirty-seven experts participated (10 countries). Twenty-nine responded to round 1 (79%), 26 round 2 (70%) and 24 round 3 (65%). Round 1 generated 79 statements outlining the interacting components of exercise prescription. Following rounds 2 and 3, consensus was achieved for 46 important components of exercise and dosage prescription across 5 themes (clinical reasoning, dosage variables, exercise variables, evaluation criteria and progression) and 2 subthemes (progression criteria and progression variables). Excellent agreement and qualitative data supports exercise prescription complexity and the need for individualised, acceptable, and feasible exercise. Only 37% of important exercise components were generated from clinical trial data. Agreement was highest (88%-96%) for 3 dosage variables: intensity of effort, frequency, and repetitions. Multiple exercise and dosage variables are important, resulting in complex and individualised exercise prescription not found in clinical trials. Future research should use these important variables to prescribe an evidence-informed approach to exercise.

378-P: Upregulation of Regenerating Gene IV and Hepatocyte Growth Factor in Cardiomyocytes by Intermittent Hypoxia and Its MicroRNA-Mediated Mechanism

Sleep apnea syndrome is characterized by recurrent episodes of oxygen desaturation and reoxygenation (intermittent hypoxia [IH]) and is a risk factor for cardiovascular disease (CVD) and insulin resistance. However, the mechanisms linking IH stress and CVD remain elusive. We exposed rat H9c2 and mouse P19.CL6 cardiomyocytes either to sustained hypoxia (1% O2 [SH]), 70 cycles/24 h of IH (5 min SH/10 min normoxia [21% O2]), mimicking SAS patients, or normoxia for 24 h, analyzed mRNA expression and found that IH significantly increased the mRNA levels of regenerating gene (Reg) IV and hepatocyte growth factor (Hgf). Reg IV and Hgf in IH-treated cell medium were also increased. In order to analyze the IH-induced expression mechanisms of Reg IV and Hgf genes, we prepared reporter plasmids containing Reg IV and Hgf upstream of luciferase reporter gene, and transfected them into P19.CL6 cardiomyocytes. The promoter activities of the genes were not increased by IH. Target mRNA search of microRNA (miR)s revealed that both the mRNAs have a potential target sequence for miR-499. The miR-499 level of IH-treated cells was significantly decreased than that of normoxia-treated cells. To clarify the role of miR-499, miR-499 mimic and non-specific control RNA (miR-499 mimic NC) were introduced into P19.CL6 cells, and the IH-induced up-regulation of Reg IV and Hgf was abolished by introduction of miR-499 mimic. We also measured apoptosis of IH/SH-stimulated P19.CL6 cells by TUNEL method and found that SH but not IH increased apoptosis and that Reg IV and Hgf in cultured medium significantly reduced. The addition of Reg IV and/or Hgf in the culture medium attenuated the SH-induced apoptosis. These results indicate that IH stress increased levels of Reg IV and Hgf mRNA via the inhibition of the miR-499-mediated mRNA degradation, leading to protect cardiomyocytes against hypoxia -induced apoptosis. Disclosure S. Takasawa: None. A. Itaya-hironaka: None. A. Yamauchi: None. M. Makino: None. S. Sakuramoto-tsuchida: None. T. Uchiyama: None. Y. Takeda: None. Y. Kyotani: None. H. Ota: None.

Cognitive Behavioral Therapy for Prodromal Stage of Psychosis-Outcomes for Transition, Functioning, Distress, and Quality of Life: A Systematic Review and Meta-analysis.

This study aimed to provide insight into the efficacy of cognitive-behavioral therapy for psychosis (CBTp) in patients with "clinical high risk of psychosis (CHR-P)". Major scientific databases were searched up to April 17, 2020. Randomized controlled trials in CHR-P individuals, comparing CBTp with needs-based interventions (NBI, including treatment as usual or nonspecific control treatment) were included, following PRISMA guidelines. The primary outcome (efficacy) was transition to psychosis by 6 months, 12 months, 24 months, and over 24 months. Secondary outcomes were change in attenuated psychotic symptoms, depression, distress, improvements in functioning, and quality of life. Ten randomized controlled studies met inclusion criteria. The comparisons included 1128 participants. CBTp was significantly more efficacious in reducing rate of transition to psychosis by 6 months (after post-hoc sensitivity analysis) (relative risk [RR] = 0.44, 95% confidence interval [CI]: 0.26, 0.73), 12 months (RR = 0.44, 95% CI: 0.30, 0.64), 12 months (RR = 0.46, 95%CI: 0.30, 0.69), and over 24 months (RR = 0.58, 95% CI: 0.35, 0.95) after treatment, compared with those receiving NBI. CBTp was also associated with more reduced attenuated psychotic symptoms by 12 months (SMD = -0.17, 95% CI: -0.33, -0.02) and by 24 months (SMD = -0.24, 95% CI: -0.43, -0.06). No beneficial effects on functioning, depression, quality of life, or distress were observed favoring CBTp. CBTp is effective in reducing both psychosis transition rates and attenuated psychotic symptoms for the prodromal stage of psychosis. It is a promising intervention at the preventative stage.

Induction of Therapeutic Protection in an HPV16-Associated Mouse Tumor Model Through Targeting the Human Papillomavirus-16 E5 Protein to Dendritic Cells

HPV E5 is an oncoprotein mainly expressed in premalignant lesions, which makes it an important target for a vaccine to prevent or cure cervical cancer (CC). In this study, we evaluated whether E5 targeted to DEC-205, present in dendritic cells (DCs), could induce a therapeutic protection against HPV16-induced tumor cells in a mouse model. The HPV-16 E5 (16E5) protein was cross-linked to a monoclonal antibody (mAb) specific to mouse DEC-205 (anti-DEC-205:16E5) or to an isotype control mAb (isotype:16E5). Rotavirus VP6 was cross-linked to the mouse anti-DEC-205 mAb (anti-DEC-205:VP6) as a non-specific antigen control. BALB/c mice were inoculated subcutaneously (s.c.) with the 16E5-expressing BMK-16/myc tumor cells, and 7 and 14 days later the mice were immunized s.c. with the conjugates, free 16E5 or PBS in the presence of adjuvant. Tumor growth was monitored to evaluate protection. A strong protective immune response against the tumor cells was induced when the mice were inoculated with the anti-DEC-205:16E5 conjugate, since 70% of the mice controlled the tumor growth and survived, whereas the remaining 30% developed tumors and died by day 72. In contrast, 100% of the mice in the control groups died by day 30. The anti-DEC-205:16E5 conjugate was found to induce 16E5-specific memory T cells, with a Th1/Th17 profile. Both CD4+ and CD8+ T cells contributed to the observed protection. Finally, treating mice that had developed tumors with an anti-PD-1 mAb, delayed the tumor growth for more than 20 days. These results show that targeting 16E5 to DEC-205, alone or combined with an immune checkpoint blockade, could be a promising protocol for the treatment of the early stages of HPV-associated cancer.

The CRISPR-Cas Mechanism for Adaptive Immunity and AlternateBacterialFunctions Fuels Diverse Biotechnologies.

Bacterial and archaeal CRISPR-Cas systems offer adaptive immune protection against foreign mobile genetic elements (MGEs). This function is regulated by sequence specific binding of CRISPR RNA (crRNA) to target DNA/RNA, with an additional requirement of a flanking DNA motif called the protospacer adjacent motif (PAM) in certain CRISPR systems. In this review, we discuss how the same fundamental mechanism of RNA-DNA and/or RNA-RNA complementarity is utilized by bacteria to regulate two distinct functions: to ward off intruding genetic materials and to modulate diverse physiological functions. The best documented examples of alternate functions are bacterial virulence, biofilm formation, adherence, programmed cell death, and quorum sensing. While extensive complementarity between the crRNA and the targeted DNA and/or RNA seems to constitute an efficient phage protection system, partial complementarity seems to be the key for several of the characterized alternate functions. Cas proteins are also involved in sequence-specific and non-specific RNA cleavage and control of transcriptional regulator expression, the mechanisms of which are still elusive. Over the past decade, the mechanisms of RNA-guided targeting and auxiliary functions of several Cas proteins have been transformed into powerful gene editing and biotechnological tools. We provide a synopsis of CRISPR technologies in this review. Even with the abundant mechanistic insights and biotechnology tools that are currently available, the discovery of new and diverse CRISPR types holds promise for future technological innovations, which will pave the way for precision genome medicine.

ImmunoPET of CD146 in Orthotopic and Metastatic Breast Cancer Models.

The overexpression of CD146 in breast cancer is considered a hallmark of tumor progression and metastasis, particularly in triple negative breast cancer. Aimed at imaging differential CD146 expressions in breast cancer, a noninvasive method for predictive prognosis and diagnosis was investigated using a 64Cu-labeled CD146-specific monoclonal antibody, YY146. CD146 expression was screened in human breast cancer cell lines using Western blotting. Binding ability was evaluated using flow cytometry and immunofluorescent staining. YY146 was conjugated with 1,4,7-triazacyclononane-triacetic acid (NOTA) and radiolabeled with 64Cu following standard procedures. Serial PET or PET/CT imaging was performed in orthotopic and metastatic breast cancer tumor models. Biodistribution was performed after the final time point of imaging. Finally, tissue immunofluorescent staining and hematoxylin and eosin (H&E) staining were performed on tumor tissues. The MDA-MB-435 cell line showed the highest CD146 expression level, whereas MCF-7 had the lowest level at the cellular level. ImmunoPET showed that MDA-MB-435 orthotopic tumors had high and clear radioactive accumulation after the administration of 64Cu-NOTA-YY146. The tumor uptake of 64Cu-NOTA-YY146 in MDA-MB-435 was significantly higher than that in MCF-7 and nonspecific IgG control groups (P < 0.01). Biodistribution verified the PET imaging results. For metastatic models, 64Cu-NOTA-YY146 allowed for the visualization of high radioactivity accumulation in metastatic MDA-MB-435 tumors, which was confirmed by ex vivo biodistribution of lung tissues. H&E staining proved the successful building of metastatic tumor models. Immunofluorescent staining verified the differential expression of CD146 in orthotopic tumors. Therefore, 64Cu-NOTA-YY146 could be used as an immunoPET probe to visualize CD146 in the breast cancer model and is potentially useful for cancer diagnosis, prognosis prediction, and monitoring therapeutic response.

Mindfulness-based programmes for mental health promotion in adults in nonclinical settings: A systematic review and meta-analysis of randomised controlled trials.

There is an urgent need for mental health promotion in nonclinical settings. Mindfulness-based programmes (MBPs) are being widely implemented to reduce stress, but a comprehensive evidence synthesis is lacking. We reviewed trials to assess whether MBPs promote mental health relative to no intervention or comparator interventions. Following a detailed preregistered protocol (PROSPERO CRD42018105213) developed with public and professional stakeholders, 13 databases were searched to August 2020 for randomised controlled trials (RCTs) examining in-person, expert-defined MBPs in nonclinical settings. Two researchers independently selected, extracted, and appraised trials using the Cochrane Risk-of-Bias Tool 2.0. Primary outcomes were psychometrically validated anxiety, depression, psychological distress, and mental well-being questionnaires at 1 to 6 months after programme completion. Multiple testing was performed using p < 0.0125 (Bonferroni) for statistical significance. Secondary outcomes, meta-regression and sensitivity analyses were prespecified. Pairwise random-effects multivariate meta-analyses and prediction intervals (PIs) were calculated. A total of 11,605 participants in 136 trials were included (29 countries, 77% women, age range 18 to 73 years). Compared with no intervention, in most but not all scenarios MBPs improved average anxiety (8 trials; standardised mean difference (SMD) = -0.56; 95% confidence interval (CI) -0.80 to -0.33; p-value < 0.001; 95% PI -1.19 to 0.06), depression (14 trials; SMD = -0.53; 95% CI -0.72 to -0.34; p-value < 0.001; 95% PI -1.14 to 0.07), distress (27 trials; SMD = -0.45; 95% CI -0.58 to -0.31; p-value < 0.001; 95% PI -1.04 to 0.14), and well-being (9 trials; SMD = 0.33; 95% CI 0.11 to 0.54; p-value = 0.003; 95% PI -0.29 to 0.94). Compared with nonspecific active control conditions, in most but not all scenarios MBPs improved average depression (6 trials; SMD = -0.46; 95% CI -0.81 to -0.10; p-value = 0.012, 95% PI -1.57 to 0.66), with no statistically significant evidence for improving anxiety or distress and no reliable data on well-being. Compared with specific active control conditions, there is no statistically significant evidence of MBPs' superiority. Only effects on distress remained when higher-risk trials were excluded. USA-based trials reported smaller effects. MBPs targeted at higher-risk populations had larger effects than universal MBPs. The main limitation of this review is that confidence according to the Grading of Recommendations Assessment, Development and Evaluation (GRADE) approach is moderate to very low, mainly due to inconsistency and high risk of bias in many trials. Compared with taking no action, MBPs of the included studies promote mental health in nonclinical settings, but given the heterogeneity between studies, the findings do not support generalisation of MBP effects across every setting. MBPs may have specific effects on some common mental health symptoms. Other preventative interventions may be equally effective. Implementation of MBPs in nonclinical settings should be partnered with thorough research to confirm findings and learn which settings are most likely to benefit.

Effects of a movement control and tactile acuity training in patients with nonspecific chronic low back pain and control impairment \u2013 a randomised controlled pilot study

BackgroundNonspecific chronic low back pain (NSCLBP) is a heterogeneous condition that is associated with complex neuromuscular adaptations. Exercise is a widely administered treatment, but its effects are small to moderate. Tailoring patient-specific exercise treatments based on subgroup classification may improve patient outcomes.ObjectiveIn this randomised controlled pilot study, our objective was to compare the feasibility and possible effects of a specific sensorimotor treatment (SMT) with those of a general exercise (GE) programme on patients with NSCLBP and control impairment (CI).MethodsPatients with NSCLBP and CI were randomised into an SMT or a GE programme spanning 6 sessions each. The feasibility criteria included the study design, assessments, interventions and magnitudes of effects, and costs. Adverse events were documented. Primary (pain, physical function, and quality of life) and secondary outcomes were assessed three times: twice at baseline (t1a and t1b) to estimate parameter stability and once after the intervention (t2).ResultsTwo-hundred and twenty-seven patients were screened to include 34 participants with NSCLBP and CI. Both treatment programmes and the assessments seemed feasible because their durations and contents were perceived as adequate. The total cost per participant was €321. Two adverse events occurred (one not likely related to the SMT, one likely related to the GE intervention).The SMT showed a tendency for superior effects in terms of pain severity (SMT t1a 3.5, t2 1.1; GE t1a 3.0, t2 2.0), pain interference (SMT t1a 1.9, t2 0.4; GE t1a 1.5, t2 0.9), physical component of quality of life (SMT t1a 39, t2 46; GE t1a 45, t2 48), and movement control.ConclusionsThe SMT approach proposed in this study is feasible and should be tested thoroughly in future studies, possibly as an addition to GE. To ensure the detection of differences in pain severity between SMT and GE in patients with NSCLBP with 80% power, future studies should include 110 patients. If the current results are confirmed, SMT should be considered in interventions for patients with NSCLBP and CI.Trial registrationRegistered in the German Register for Clinical Trials (Trial registration date: November 11, 2016; Trial registration number: DRKS00011063; URL of trial registry record); retrospectively registered.