This project tries to artificially grow one of the most ornamentally significant Petunia hybrida when it is out of season. Murashige and Skoog Basal Medium (MS Basal Medium) was used for in vitro growth of P. hybrida, supplemented with different Plant Growth Regulators (PGRs). Various physical and chemical factors on in vitro growth of P. hybrida using different explants were analyzed. The type of auxin that proved significant for somatic embryogenesis of P. hybrida Vilm was 2,4-Dichlorophenoxyacetic acid (2.0 mg/L) using the leaf as explant under controlled environmental conditions, i.e., 92% at 23oC with a 5.8 pH and 16 hours photoperiod & 30g/l sucrose in. Friable calli were also developed from leaf, node, and internodal explants on a tissue culture medium (MS basal medium) when supplemented with other PGRs in certain combinations i.e. 2,4-Dichlorophenoxyacetic acid (2.0 mg/L), Benzyl Amino Purine (2.5mg/L), Benzyl Amino Purine + 2,4-Dichlorophenoxyacetic acid (2mg/L +2.5mg/L), Naphthalene Acetic Acid (1.5mg/L), 2,4-Dichlorophenoxyacetic acid + Naphthalene Acetic Acid (2mg/L+1.5mg/L) and Benzyl Amino Purine + Naphthalene Acetic Acid (2.5mg/L+0.5mg/L) at optimal physical factors under strict aseptic environment. These results may enable one to produce the required type of P. hybrida Vilm during off and on seasons on a commercial scale by using plant tissue culture as a tool.

In this study, an innovative analytical analysis of fractional-order partial differential equations is presented by Shehu transformation method. Fractional-order differential equations provide the useful dynamics of phys- ical systems and thus provide novel and efficient information about given physical systems. In this study, Shehu transform is used to create an approximate analytical solution through the time-fractional partial differential equations (system of equations) with the Adomian decomposition method and Variational iter- ation transform method along with Shehu transformation. Laplace and Sumudu transformation have been refined to form Shehu transformation. An algorithm is established for expressing the Shehu transform for the fractional operators like Riemann-Liouville and Caputo by using this new integral transform. Higher-order fractional differential equations are solved in the Caputo sense. Shehu transformation is used to simplify the problems before implementing the decomposition and variational iteration methods to achieve the problem’s comprehensive solutions. This method provides a series form solution with easily computed components and a higher rate of convergence to the exact solution of the targeted problem. The reliability of this process is demonstrated through physical problems. MATLAB software is used to analyze the problems graphically. It is observed that integer-order differential equations do not properly model various phenomena in different fields of science and engineering in relation to fractional-order differential equations. This method is simple and accurate analytical technique that can solve other partial differential equations of fractional order as well.

Background The role of cancer rehabilitation is key in maintaining functional abilities and improving quality of life. Rehabilitation is a component of the CureALL program, which aims to achieve at least 60% survival for children with cancer globally while ensuring that suffering is reduced for every child with cancer. The demand for pediatric cancer rehabilitation will increase as the life expectancy of these children improves, but not all countries in Latin America are prepared. With the support of Sociedad Latinoamericana de Oncologia Pediatrica (SLAOP), the aims were (1) identify the availability of rehabilitation services for children and adolescent with cancer in Latin America; (1.1) identify the rehabilitation services and the comprehensive approach they have (physical, psychological, cognitive, social); (1.2) identify the rehabilitation healthcare professional that work with pediatric oncology; and (2) explore the barriers of access and opportunity that exists in Latin-America regarding rehabilitation of children and adolescents with cancer. Methods A survey was sent to all members of the 21 countries of SLAOP. The survey had questions regarding different aspects of rehabilitation, access, and barriers. Results Out of the 21 countries who are members of SLAOP, we had responses from 19 with a total of n=123 responses. Most healthcare providers (98.4%) know the importance of rehabilitation in pediatric oncology, and 86.2% had rehabilitation teams. Only 19 respondents said they had pediatric oncology rehabilitation educational programs. Barriers to access rehabilitation were reported by 83% providers: Approachability (n=25), Acceptability (n=4), Availability (n=76), Affordability (n=35), Appropriateness (n=24) The main barriers associated with availability were distance (n=31) and lack of health care professionals (n=39). Conclusions While the importance of pediatric oncology rehabilitation is known in Latin America, there are many factors that act as barriers to access to care for children and adolescents with cancer. It is key to address these barriers to improve the quality of life of Latin American pediatric oncology patients.

Objective We aimed to describe the clinical outcomes of patients receiving veno-arterial extracorporeal membrane oxygenation therapy considering clinical context and pH at cannulation. Methods We reviewed all patients having received veno-arterial extracorporeal membrane oxygenation therapy at a tertiary referral center during the 2005-2020 period with 1-year complete follow up. Our cohort was divided in three groups according to the pH level at cannulation: pH<7 (group 1), pH 7-7.2 (group 2) and pH>7.2 (group 3). Survival was analyzed using Kaplan–Meier method. Association between pH group and survival was estimated using a Cox model. Results Among the 951 patients in our database, 572 were included in 3 different groups according to their pH at implantation: 60 patients in group 1, 115 in group 2 and 397 in group 3. Main indications of mechanical support were refractory cardiogenic shock (36%), post cardiotomy (28%), early graft failure (12%), refractory cardiac arrest (11%). One-year survival rate was 13% in group 1, 36% in group 2, 43% in group 3 respectively (p<0.001). Death mainly occurred within the first month. The strong correlation between pH and lactates led to propose a simple “three seven rule”: pH<7 and lactate >7 was associated with <7% survival. Conclusion Extracorporeal veno-arterial membrane oxygenation should be considered with caution in patients with pH<7. Lactates and pH level might be important parameters to elaborate a new score to predict survival in this population. The simplicity of the “three seven rule” can be very relevant when facing emergency situations.

Background: Allergen immunotherapy (AIT) may have a long-term disease modifying effect. The aim of this study is to demonstrate the long-term benefit of MCT®-associated allergoid pollen SCIT (MCT®-associated -AIT) on allergic rhinitis (AR) and asthma in clinical practice. Methods: In this retrospective Real-World-Evidence (RWE) study the impact of AIT on the progression of AR and onset of need for asthma medication was analyzed using a German longitudinal database. Anonymized prescription data of AR patients and exactly matched control patients aged from 5-65 years were analyzed. Results: Significantly less patients treated with MCT®-associated-AIT did receive prescriptions for symptomatic AR medication in the follow up period vs. control group (OR: 0.27; p < 0.001). Further, significantly less asthmatic patients under MCT®-associated-AIT did receive prescriptions for asthma medications (OR: 0.48; p = 0.004). In addition, the prescriptions of AR and asthma medication for MCT®-associated-AIT patients were significantly reduced in the follow-up vs. baseline and control group (24.2% and 35.6%, respectively, p < 0.001).The probability of asthma medication onset in non-asthmatic patients during follow-up was significantly reduced for AIT patients compared to controls (OR: 0.77, p = 0.001). All endpoints were significant for children/adolescents and adults in the individual analyses. Conclusions: This study gives evidence for long-term benefits up to 9.5 years of MCT®-associated-AIT on the need for AR and new-onset asthma medication in AR patients and asthma medication in asthmatics in an RWE setting.

In this paper, the relationship between the conveying pressure drop and the gas velocity were firstly characterized based on pressure signal analysis. The critical gas velocity was obtained to classify the transfer bottle into stationary station and motion state, and pressure fluctuation characteristics in each state were determined. Further, derived from the traditional additional pressure drop method, the model for pressure drop prediction was developed. The gas pressure drop was further divided into four terms, including those in the conveying pipe, through the sudden contraction from the pipe to the head of the transfer bottle, through the sudden expansion from the end of the transfer bottle to the pipe, and through the gap between the bottle and the pipe. The solid-phase friction pressure drop was determined based on correctly estimation of the dynamic friction coefficient.The model can satisfactorily predict the pressure drop, giving errors mostly smaller than ±10%.

Objectives: New temperature management concepts of moderate and mild hypothermic circulatory arrest during aortic arch surgery have gained weight over profound cooling. Comparisons of all temperature levels have rarely been performed. We performed direct and indirect comparisons of deep hypothermic circulatory arrest (DHCA) (≤20°C), moderate hypothermic circulatory arrest (MHCA) (20.1°C to 25°C), and mild hypothermic circulatory arrest (mild HCA) (≥25.1°C) in a network meta-analysis. Methods: The literature was systematically searched for all papers published through February 2022 reporting on clinical outcomes after aortic arch surgery utilizing DHCA, MHCA and mild HCA. The primary outcome was operative morality. The secondary outcomes were postoperative stroke and acute kidney failure (AKI). Results: A total of 34 studies were included, with a total of 12 370 patients. DHCA was associated with significantly higher postoperative incidence of stroke when compared with MHCA (odds ratio (OR), 1.46, 95% (confidence interval) CI, 1.19-1.78) and mild HCA: (OR, 1.50, 95% CI, 1.14-1.98). Furthermore, DHCA and MHCA were associated with higher operative mortality when compared with mild HCA (OR 1.71, 95% CI, 1.23-2.39 and OR 1.50, 95% CI, 1.12-2.00, respectively). Separate analysis of randomized and propensity score matched studies showed sustained increased risk of stroke with DHCA in contrast to MHCA and mild HCA (OR, 1.61, 95% CI, 1.18-2.20, P-value = 0.0029 and OR, 1.74, 95% CI, 1.09-2.77, P-value = 0.019). Conclusions: In the included studies, the moderate to mild hypothermia strategies were associated with decreased operative mortality and the risk of postoperative stroke. Large-scale prospective studies are warranted to further explore appropriate temperature management for the treatment of aortic arch pathologies.

Background: Primary cardiac angiosarcoma is a rare cardiac tumor. Most of them grew in the right atrium, often invading the superior vena cava and pericardium. The clinical symptoms of majority patients may present right heart failure, superior vena cava obstruction, and even pericardial tamponade. Case presentation: In this case report, we presented a 76-year-old man presented with chest tightness, fatigue, pericardial effusion, and arrhythmia. The transthoracic echocardiography (TTE) showed that a mass in the right atrium. Histopathology and immunohistochemistry confirmed the diagnosis of angiosarcoma. The patient was discharged from hospital after surgery. Conclusions: For the patients with early stage cardiac angiosarcoma without evidence of metastasis, radical resection of the primary tumor remains the most important approach for optimal survival.

A 65-year-old female presented to our center with exhaustion and shortness of breath after daily exercise for two months. On physical examination, vital signs were stable. All laboratory results were unremarkable. Transthoracic Echocardiogram showed a round hypoechoic mass (2.3cm  2.6cm) under the tricuspid valve with clear margins, part of which closely connected with the right ventricular wall (Figure 1 A) and there was no blood flow signal inside (Figure 1 B). The mass does not communicate with the heart’s chambers and can affect blood flow in the inflow tract of the right ventricle. Contrast-enhanced computed tomography revealed a low-density round mass in the right ventricle, and it does not enhance after the injection of intravenous contrast media (Figure 1 C). The patient underwent right ventricular mass resection under cardiopulmonary bypass. The tricuspid valve was suspended during the operation to expose the cyst below the tricuspid valve. The cyst wall is almost transparent (Figure 1D). Approximately 7 ml of pellucid fluid withdraw with a syringe from the cyst (Figure 1 E1). Then the exposed cyst wall was then wholly resected (Figure 1 E2), but part of the cyst wall is the wall of the right ventricle, this part cannot be removed. The patient had an uneventful postoperative recovery. Histopathological examination revealed that the cystic wall was covered by low cuboidal epithelium, and the cystic wall was glassy fibrous tissue, consistent with cystic changes (Figure 1 F). The right ventricular watery cyst is a rare intracardiac cyst that can easily be mistaken for a myxoma. Whether such cysts will recur is worthy of our long-term follow-up attention.

A document by Sarbeswar Mandal. Click on the document to view its contents.

Recent evidence has shown that G protein-coupled receptors (GPCRs) are direct sensors of the autophagic machinery and οpioid receptors regulate neuronal plasticity and neurotransmission with an as yet unclarified mechanism. Using in vitro and in vivo experimental approaches, this study aims to clarify the potential role of autophagy and κ-opioid receptor (κ-OR) signaling on synaptic structure and integrity. We hereby demonstrate that the selective κ-OR agonist U50,488H, induces autophagy in a time-and dose-dependent manner in neuronal cells by upregulating microtubule-associated protein Light Chain 3-II (LC3-II), Beclin 1 and Autophagy Related Gene 5 (ATG5). Pretreatment of neuronal cells with pertussis toxin blocked the above κ-OR-mediated cellular responses. Our molecular analysis also revealed a κ-OR-driven upregulation of becn1 gene through ERK1,2-dependent activation of the transcription factor CREB in neuronal cells. Moreover, our studies demonstrated that sub-chronic U50,488H administration in mice causes profound increases of specific autophagic markers in the hippocampus with a concomitant decrease of several pre- and post-synaptic proteins such as spinophilin, postsynaptic density protein 95 (PSD-95) and synaptosomal associated protein 25 (SNAP25). Finally, using acute stress, a stimulus known to increase the levels of the endogenous κ-OR ligand dynorphin, we are demonstrating that administration of the κ-ΟR selective antagonist, nor-binaltorphimine (norBNI), blocks the induction of autophagy and the stress-evoked reduction of synaptic proteins in the hippocampus. These findings provide novel insights about the essential role of autophagic machinery into the mechanisms through which κ-OR signaling regulates brain plasticity.

Background: The biological process of aging is brought on by a variety of variables and mechanisms, among which, the most comprehensive is the aging process caused by oxidative stress. The theory of the “homology of medicine and food”, originated from the thinking of traditional Chinese medicine, has gained popularity in recent years, and is believed to be utilized as a dietary therapy to ease the symptoms of aging. Purpose: We searched the recent 10-year literature on the “homology of medicine and food” traditional Chinese medicine to alleviate aging and prolong life, and summarized its anti-aging mechanism under the influence of oxidative stress. Experimental approach: Six electronic databases, including China National Knowledge Infrastructure (CNKI), Wanfang database, Chinese Scientific Journals Database (VIP), PubMed, EMBASE, and Cochrane Library, were searched from June 2012 to June 2022 for identifying eligible studies. Key Results: The “homology of medicine and food” traditional Chinese medicine mainly scavenges free radicals, alleviates mitochondrial DNA damage, regulates telomeres and telomerase, regulates nutrient and energy sensing signaling, up-regulates sirtuins signaling pathway, and activates the Keap1-Nrf2/ARE signaling pathway mechanisms delay aging and prolong lifespan, and expect to find microcirculation mechanisms and new targets. Conclusions & Implications: The “homology of medicine and food” traditional Chinese medicine relieves the aging of the body and cells under oxidative stress through 6 pathways and is expected to find the correlation between the microcirculation formation mechanisms and identify new research targets and factors, which will provide new opportunities for life extension, prevention, and treatment of age-related diseases.

Over the last two decades, there has been a huge increase in our understanding of microbial diversity, structure and composition enabled by high throughput sequencing (HTS) technologies. Yet, it is unclear how the number of sequences translates to the number of cells or species within the community. Additional observational data may be required to ensure relative abundance patterns from sequence reads are biologically meaningful or presence absence data may be used instead of abundance. The goal is to obtain robust community abundance data, simultaneously, from environmental samples. In this issue of Molecular Ecology Resources, Karlusich et al., (2022) describe a new method for quantifying phytoplankton cell abundance. Using Tara Oceans datasets, the authors propose the photosynthetic gene psbO for reporting accurate relative abundance of the entire phytoplankton community from metagenomic data. The authors demonstrate improved correlations with traditional optical methods including microscopy and flow cytometry, improving upon current molecular identification typically using rRNA markers genes. Furthermore, to facilitate application of their approach, the authors curated a psbO gene database for accessible taxonomic queries. This is an important step towards improving species abundance estimates from molecular data and eventually reporting of absolute species abundance, enhancing our understanding of community dynamics.

Cardiovascular and metabolic diseases (CVMDs) are a leading cause of death worldwide and imposes a huge socioeconomic burden on individuals and healthcare systems, underscoring the urgent need to develop new drug therapies. Developmental endothelial locus-1 (DEL-1) is a secreted multifunctional domain protein that can bind to integrins and play an important role in the occurrence and development of various diseases. Recently, DEL-1 has attracted great interest for its pharmacological role in the treatment and/or management of CVMDs. In this review, we present the current knowledge on the predictive and therapeutic role of DEL-1 in a variety of CVMDs, such as atherosclerosis, hypertension, cardiac remodeling, ischemic heart disease, obesity, and insulin resistance. Collectively, DEL-1 is a promising biomarker and therapeutic target for CVMDs.

Background and purpose: Our previous work proved that HMGB1 A Box significantly protected TH+ neurons in Parkinson’s disease (PD) model mice and inhibited microglia activation and T cell infiltration including Th17 in the substantia nigra (SN). This study explored the mechanism of Th17 differentiation and how A Box inhibiting this process. Experimental approach: Clodronate liposomes were used to deplete the peripheral monocytes of mice, infused the labeled CD3+ T cells, and used immunoprecipitation to knock down the primary cells HMGB1 in vitro. At the same time, we combined the data of serum and blood cells of PD patients to study. Key results: Depletion of peripheral monocytes/macrophages reduced Th17 cell infiltration in the SN of MPTP mice and protected TH+ neurons. Co-culture experiments with knockdown of HMGB1 in primary cells showed that HMGB1, which induces monocyte/macrophage migration and infiltration into the SN, originates from neurons rather than glial cells. Data from MPP+-treated midbrain cell models and assays associated with adoptive transfer of CD3+ cells suggest that monocyte/macrophage and T cell migration into the SN is mediated by HMGB1/CXCL12-CXCR4. Co-immunoprecipitation and immunofluorescence confirmed that HMGB1 A Box bind to CXCR4 on T cells and macrophages, thereby competitively inhibiting their infiltration in SN. The HMGB1/CXCL12 complex is also present in the serum of PD patients. Conclusions and implications: HMGB1 A Box protects TH+ neurons by binding CXCR4 to inhibit the migration / infiltration of T cells and macrophages to SN mediated by HMGB1 / CXCL12 complex formed by neuron derived HMGB1.

Pena-Shokeir Syndrome is considered to be a fatal congenital condition that is rarely diagnosed in neonates. We present the first ever reported case of Pena-Shokeir Syndrome from Syria. Both clinical assessment and early prenatal diagnosis are necessary to give more realistic options for the mother and the baby.

Pain can be a challenging complication experienced by patients who have Guillain- Barré Syndrome(GBS). Managing pain can be difficult and may require the use of multiple agents and still may not treat pain. This case report describes the use of an epidural to manage pain in a patient with GBS.

Background and Purpose: ISX9 is a neurogenesis-promoting small molecule compound which can upregulate the expression of NeuroD1 and induce differentiation of neuronal, cardiac and islet endocrine progenitors. So far, the molecular mechanisms underlying the action of ISX9 still remain elusive. Experimental Approach: To identify a novel agonist of the Wnt/β‐catenin, a cell-based SuperTOPFlash reporter system was used to screen known-compound libraries. An activation effect of ISX9 on the Wnt/β‐catenin pathway was analysed with the SuperTOPFlash or SuperFOPFlash reporter system. Effects of ISX9 on Axin1/LRP6 interaction were examined using a mammalian two-hybrid system, co‐immunoprecipitation, microscale thermophoresis (MST), emission spectra and mass spectroscopy assays. The expression of Wnt target and stemmness marker genes were evaluated with real‐time PCR and immunoblotting. In vivo hair regeneration abilities of ISX9 were analysed by immunohistochemical staining, real‐time PCR and immunoblotting in hair regrowth model using C57BL/6J mice. Key Results: In this study, ISX9 was identified as a novel agonist of the Wnt/β‐catenin pathway. ISX9 targeted Axin1 by covalently binding to its N-terminal region and potentiated the LRP6-Axin1 interaction, thereby resulting in the stabilization of β‐catenin and upregulation of Wnt target genes and stemmness marker genes. Moreover, the topical application of ISX9 markedly promoted hair regrowth in C57BL/6J mice and induced hair follicle transition from telogen to anagen via enhancing Wnt/β‐catenin pathway. Conclusions and Implications: Taken together, our study unraveled that ISX9 could activate Wnt/β‐catenin signaling by potentiating the association between LRP6 and Axin1, and may be a promising therapeutic agent for alopecia treatment

The analysis of genomic data can be an intimidating process, particularly for researchers who are not experienced programmers. Commonly used analyses are spread out across programs, each of which require their own input formats, and data must often be wrangled and re-wrangled into new formats to split the data according to categorical metadata variables, such as population or family. Here, we introduce snpR, and R package that allows for user-friendly processing of SNP genomic data by automating data sub-setting and processing across categorical metadata, integrating approaches contained in many different packages under a single ecosystem, and allowing for iterative, efficient analysis focused on a single R object across an entire analysis pipeline.

Meteorological simulations using hydrological models are now important for water and environmental resource management. The hydrological system of a catchment cannot be measured precisely in practice. A combination of limited measurements available in space, as well as a need to extrapolate information from available measurements in time, led to the use of the hydrological model. The aim of the study was to assess the catchment retention capacity of the flysh catchments of the Smugawka (19°47-49°33) and Mszanka (20°15-49°44′) rivers located in the Beskid Wyspowy, southern Poland. Due to the large heterogeneity of the catchment area and the limitations of the measurement technique, SWAT+ was employed in our study. Forested and pasture areas had higher soil water retention potential, while lower soil retention potential values are correlated with agricultural areas. Surface runoff of both catchments correlated negatively with soil water retention potential, evapotranspiration, and positively with mean annual precipitation. Soil moisture was related to electrical conductivity and inversely correlated with mean annual precipitation. A study has shown that SWAT+ can be applied to calculate the mean annual continuous evapotranspiration for meadows, forests, and agricultural areas. Curve number may be used as a tool for water retention in floodplains. In line with the physical-based SWAT+, the Landscape Hydric Potential (LHP) index can be incorporated into the existing meteorological data models. In the future, watershed models based on water balance should be developed to increase resilience in the flysh Carpathian mountains to climate change.

Objective- Assessment of lipid profile and atherogenic lipid indices in children with transfusion dependent thalassemia (TDT) and to compare them with matched healthy children. Method- The study group consisted of total 72 patients of TDT aged between 3-14 years while the control group had 83 age and sex matched healthy children. Estimation of fasting lipid profile and lipid indices - atherogenic index of plasma (AIP), Castelli risk index I and II, atherogenic coefficient were calculated and compared between the two groups. Result – Compared to the control group, the mean LDL, HDL, and cholesterol level were significantly lower among case group (P value < 0.001). The mean VLDL and triglycerides were significantly higher in case group (P value < 0.001). Lipid indices including atherogenic index of plasma (AIP), Castelli risk index I and II, atherogenic coefficients were significantly higher in TDT children. Conclusion- Dyslipidemia and increased risk of atherosclerosis were found in children of TDT as they had elevated atherogenic lipid indices. Our study underlines the importance of routine use of these indices in TDT children. Future studies should focus on lipid indices in this high group of children so that preventive strategies can be planned accordingly.

Aim Assess the effectiveness and safety of nivolumab versus cetuximab in patients with R/M HNSCC, as well as to analyze possible prognostic factors for response to treatment with nivolumab. Methods We conducted an observational, retrospective, descriptive study of patients with R/M HNSCC who initiated treatment with nivolumab or cetuximab monotherapy in two periods of equivalent duration. Overall efficacy was measured in progression-free survival (PFS) and overall survival (OS); safety was evaluated using the CTCAE (Common Terminology Criteria for Adverse Events) classification version 5.0 of the National Cancer Institute (NCI). Results Median overall survival (OS) was 9.1 months with nivolumab (n=34) vs. 6.3 months with cetuximab (n=12)(HR=0.5; 95%CI: 0.24-1.03; p=0.058). Progression free survival (PFS) were 4.3 for nivolumab and 4.65 months for cetuximab (HR=0.59; 95%CI: 0.29-1.19; p=0.14). Any grade adverse events (AEs) were reported in 97% and 100% of the patients treated with nivolumab and cetuximab. Serious AEs were observed in 26% and 58% of the patients respectively. Elevated albumin values, lymphocytosis, neutropenia and elevated neutrophil/lymphocyte ratio values have positive prognostic value on the response to nivolumab in R/M CCECC. Conclusion Effectiveness of nivolumab in terms of OS remains superior to cetuximab. OS, PFS and severe or any grade AEs were superior in both arms of our study than in the clinical trials. The AEs profile of nivolumab differs in our study from the clinical trials’ observations. We have identified four positive statistically significant prognostic variables on the response to nivolumab in R/M HNSCC.

Background: Natural killer cells are critical immune cells for AML targeting. However, little is known about the relationship between using checkpoint inhibitors and Hsp70 as NK cell activators and later immune responses to control AML. We aimed to investigate the antitumor effects of NK cells pre-treated with ex-vivo Hsp70, human PD-1 blocker, and IL-15. Procedure: The NK cells were isolated from patients-derived MNCs using MACS and activated using the different combinations of Hsp70, PD-1 blocker, and IL-15. Then their killing potential and the expression pattern of PRF-1, PIK3CB, PD-1, AKT-1, FAS-L, TRAIL, and GER A & B were estimated. Results: Our data revealed that the PD-1 expression was significantly reduced after NK cell activation with the different formulas of NK cell activators. Also, the expression of NKG2A was reduced, particularly in the IL-15 and IL-15 + PD-1 blocker treated groups, and adding the Hsp70 increased its expression. Moreover, the cytotoxic effect of NK cells increased in all groups, especially in IL-15 + PD-1 blocker group, in parallel with increasing in IFN-γ releasing, Granzymes, and perforin expression. All changes in IL-15 + PD-1 blocker groups were associated with the up-regulation of PIK3CB and AKT-1 as key factors of NK cell activation. The presence of Hsp70 reduced IFN-γ releasing and down-regulation of PIK3CB, AKT-1, Granzymes, and perforin. Conclusions: We suggested that combining IL-15 and PD-1 blockers could enhance the killing potential of AML-NK cells. Moreover, Hsp70, in combination with IL-15 and PD-1 blocker, interferes activation of AML-NK cells through unknown mechanisms.

1. Given the sharp increase in agricultural and infrastructure development and the paucity of widespread data available for making conservation management decisions, a more rapid and accurate tool for identifying fish fauna in the world’s largest freshwater ecosystem, the Amazon, is needed. 2. Current strategies for identification of freshwater fishes require high levels of training and taxonomic expertise for morphological identification or genetic testing for species recognition at a molecular level. 3. To overcome these challenges, we built an image masking model (U-Net) and a convolutional neural net (CNN) to classify Amazonian fish in photographs. Fish used as training data were collected and photographed in tributaries in seasonally flooded forests of the upper Morona River valley in Loreto, Peru in 2018 and 2019. 4. Species identifications in the training images (n = 3,068) were verified by expert ichthyologists. These images were supplemented with photographs taken of additional Amazonian fish specimens housed in the ichthyological collection of the Smithsonian’s National Museum of Natural History. 5. We generated a CNN model that identified 33 genera of fishes with a mean accuracy of 97.9%. Wider availability of accurate freshwater fish image recognition tools, such as the one described here, will enable fishermen, local communities and community scientists to more effectively participate in collecting and sharing data from their territories to inform policy and management decisions that impact them directly.