Background and Purpose: In major depressive disorder (MDD), exploration of biomarkers will be helpful in diagnosing the disorder as well as in choosing a treatment, and predicting the treatment response. Currently, tRNA-derived small ribonucleic acids (tsRNAs) have been established as promising non-invasive biomarker candidates that may enable a more reliable diagnosis or monitoring of various diseases. Herein, we aimed to explore tsRNA expression together with functional activities in MDD development. Experimental Approach: Serum samples were obtained from patients with MDD and healthy controls, and small RNA sequencing (RNA-Seq) was used to profile tsRNA expression. Dysregulated tsRNAs in MDD were validated by quantitative real-time polymerase chain reaction (qRT-PCR). The diagnostic utility of specific tsRNAs and the expression of these tsRNAs after antidepressant treatment was analyzed. Key Results: In total, 38 tsRNAs were significantly differentially expressed in MDD samples relative to healthy individuals (34 upregulated and 4 downregulated). qRT-PCR was used to validate the expression of six tsRNAs that were upregulated in MDD (tiRNA-1:20-chrM. Ser-GCT, tiRNA-1:33-Gly-GCC-1, tRF-1:22-chrM.Ser-GCT, tRF-1:31-Ala-AGC-4-M6, tRF-1:31-Pro-TGG-2, and tRF-1:32-chrM. Gln-TTG). Interestingly, serum tiRNA-Gly-GCC-001 levels exhibited an area under the ROC curve of 0.844. Moreover, tiRNA-Gly-GCC-001 is predicted to suppress brain-derived neurotrophic factor (BDNF) expression. Furthermore, significant tiRNA-Gly-GCC-001 downregulation was evident following an eight-week treatment course and served as a promising baseline predictor of patient response to antidepressant therapy. Conclusion and Implications: Our current work firstly found that tiRNA-Gly-GCC-001 is a promising MDD biomarker candidate that can predict patient responses to antidepressant therapy.

Enabling real-time monitoring and control of the biomanufacturing processes through product quality insights continues to be an area of focus in the biopharmaceutical industry. The goal is to manufacture products with the desired quality attributes. To realize this rigorous attribute-focused Quality by Design (QbD) approach, it’s critical to support the development of processes that consistently deliver high-quality products and facilitate product commercialization. Time delays associated with off-line analytical testing can limit the speed of process development. Thus, developing and deploying analytical technology is necessary to accelerate process development. In this study, we have developed the Micro Sequential Injection (µSI) process analyzer and the Automatic Assay Preparation Platform (A2P2) system. These innovations address the unmet need for an automatic, online, real-time sample acquisition and preparation platform system for in-process monitoring, control, and release of biopharmaceuticals. These systems can also be deployed in laboratory areas as an off-line analytical system and on the manufacturing floor to enable rapid testing and release of products manufactured in a GMP environment.

Agitated saline microbubbles (MBs) is a common contrast agent for determining right-to-left shunt (RLS) by the contrast Transcranial Doppler (c-TCD). The size of the generated bubbles is not standardized in clinical practice. The microbubbles were generated using the recommended manual method by reciprocating motion through two syringes. The bubble size distributions (BSD) were measured using the microscopic shadow imaging technique. The results show that the diameter of MBs is mainly distributed between 10-100 µm, the mean bubble size is between 21-34 μm, the Sauter mean diameter (D 32) is mainly between 50-300 μm, and the standard deviation (SD) is between 6-17 μm in 60 experiments. It provides a more accurate basis for the recommended manual method instability. The high variance values of the BSD indicate that the manual method has low stability and repeatability. The results of this study can be useful for further improvement of the reliability of c-TCD in detecting RLS. Research Highlights This study provided the first detailed descriptions of the size distribution of microbubbles in a flowing contrast agent by the microscopic shadow imaging technique. It reveals significant differences in the bubble size of manual foaming during repeated manipulations for each individual and between individuals.

Heparin-induced Thrombocytopenia with Normal Platelet Count: A Cautionary Tale of Delayed DiagnosisAhmed Elkheshen MD1, Marawan Elmassry MD1*, Sierra Sullivan MD1, Ricardo Franco MD1, Abbie Evans MD1, Kavya Bharathidasan MD1, Arunee Motes MD1, Sameer Islam PhD3, Ebtisam Attaya Islam PhD21-Department of Internal Medicine, Texas Tech university Health Sciences Centre, Lubbock, TX, USA.2- Department of Pulmonology and Critical Care Medicine, Texas Tech university Health Sciences Centre, Lubbock, TX, USA.3- Department of Gastroenterology and Hepatology, Texas Tech university Health Sciences Centre, Lubbock, TX, USA.*Corresponding Author: Marawan Elmassry MDTexas Tech University Health Sciences Centre at Lubbock, Texas, USA.3601 4th street, Lubbock, TX 79430.Telephone/ Fax: +1-806-773-2831E-mail: [email protected] of interest: The authors have no financial conflicts to disclose.Patient’s consent: Written informed consent was obtained from the patient to publish this report in accordance with the journal’s patient consent policy.Keywords :HIT, Heparin, Thrombocytopenia, Heparin Induced Thrombocytopenia, Platelet Count.

Objective: To evaluate both short and mid-term effects of maternal COVID-19 on cardiac functions of fetuses and children. Methods: The present case-control study was conducted on 36 pregnant women who had COVID-19 infection in the second trimester of pregnancy and 30 pregnant women as healthy controls. Fetal, neonatal and infant cardiac functions were compared between the groups. Assessment of fetal cardiac functions were performed in the last trimester of the pregnancy at least 6 weeks after the recovery of infection. The first postnatal echo was performed within the first two weeks and the follow-up (second) echo was performed in the 6-8 weeks of life. Results: The demografic data were similar between groups. Interventricular septum and left ventricular posterior wall end-diastolic dimensions were significantly higher in the study group in both fetal, neonatal and infant periods. Impaired diastolic functions of right and left ventricles were detected and myocardial performance indexes with tissue doppler imaging of both lateral walls and septum were significantly higher than controls at all periods. Conclusion: Maternal COVID-19 seems to have a global impact on cardiac functions of babies in the short and mid-term periods after maternal recovery.

A rare presentation of an adult with un-diagnosed coarctation of the aorta: A case report

Leukemoid Reaction in a young woman following a TAH-BSO with severe hemorrhage: A case report

Introduction Vascular anomalies are alterations in angiogenesis and endothelial proliferation, respectively, that can generate not only aesthetic problems, but also functional disorders in fetuses and newborns. Other alterations in the regression of blood vessels can generate vascular malformations such as persistent left superior vena cava. Case report We present the case of a

The stiffness of the tumor microenvironment (TME) is dynamic and drives metabolic reprogramming in cancer cells as a consequence of tumor progression. To demonstrate the possibility to modulate the mechano-metabolomic profile of breast cancers by tuning the mechanical property and dimensionality of extracellular matrices (ECMs), we cultured triple-negative MDA-MB-231 and luminal MCF-7 cells on 2D and in 3D hydrogels based on tyramine functionalized hyaluronic acid (HTA). Using high-throughput metabolomics analyses, we established that we can differentially regulate breast cancer mechano-metabolome. The stiff hydrogels resulted in upregulated lipid and amino acid metabolism along with increasing malignancy and chemoresistancy. Reprogramming in glucose metabolism is primarily observed in cells seeded on 2D hydrogels, whereas modifications in amino acid metabolism is predominant in cells embedded in 3D stiff hydrogels. These findings suggest that matrix stiffness and dimensions have decisive roles in reprogramming breast cancer metabolome, which is the hallmark of breast cancer development and progression.

Background: Patients with AF and likelihood of bleeding can undergo left atrial appendage occlusion (LAAO) as an alternative method of stroke prophylaxis. Short-term anti-thrombotic drugs are used post-procedure to offset the risk of device-related thrombus, evidence for this practice is limited. Objectives: To investigate optimal post-implant antithrombotic strategy in high bleeding-risk patients. Methods: Patients with AF and high-risk for both stroke and bleeding undergoing LAAO were advised their peri-operative drug therapy by a multi-disciplinary physician panel. Those deemed to be at higher risk of bleeding from anti-thrombotic drugs were assigned to minimal treatment with no antithrombotics or aspirin-alone. The remaining patients received standard care (STG)with a 12week course of dual-antiplatelets or anticoagulation post-implant. We compared mortality, device-related thrombus, ischemic stroke and bleeding events during the 90 days post-implant and long-term. Event-free survival was assessed using Kaplan-Meier survival analysis, with logrank testing for statistical significance. Results: 75 pts underwent LAAO of whom 63pts(84%) had a prior serious bleeding event. The 42pts on minimal treatment were older(74.3±7.7vs71.2±7.2) with higher HASBLED score (3.6±0.9vs3.3±1.2) than the 33pts having standard care. There were no device-related thrombi or strokes in either group at 90 days post-procedure; STG had more bleeding events (5/33vs0/42,p=0.01) with associated deaths (3/33vs0/42,p=0.05). During long-term follow up (median 2.2yrs), all patients transitioned onto no antithrombotic drugs (43pts(61%)) or a single-antiplatelet (29pts(39%)). There was no evidence of early minimal treatment adversely affecting long-term outcomes. Conclusions: Short-term anti-thrombotic drugs may not be needed after LAAO implant in patients with high bleeding risk and could be harmful. Larger, prospective studies would be warranted to test these findings.

Quantitative risk assessments of chemicals are routinely performed in rodents; however, there is growing recognition that non-animal approaches can be human-relevant alternatives. There is an urgent need to build confidence in non-animal alternatives given the international support to reduce the use of animals in toxicity testing where possible. In order for scientists and risk assessors to prepare for this paradigm shift in toxicity assessment, standardization and consensus on in vitro testing strategies and data interpretation will need to be established. To address this issue, an Expert Working Group (EWG) of the 8th International Workshop on Genotoxicity Testing (IWGT) evaluated the utility of quantitative in vitro genotoxicity concentration-response data for risk assessment. The EWG first evaluated available in vitro methodologies and then examined the variability and maximal response of in vitro tests to estimate biologically relevant values for the critical effect sizes considered adverse or unacceptable. Next, the EWG reviewed the approaches and computational models employed to provide human-relevant dose context to in vitro data. Lastly, the EWG evaluated risk assessment applications for which in vitro data are ready for use and applications where further work is required. The EWG concluded that in vitro genotoxicity concentration-response data can be interpreted in a risk assessment context. However, prior to routine use in regulatory settings, further research will be required to address the remaining uncertainties and limitations.

The use of hyperinsulinemia-euglycemia therapy in the treatment of amlodipine overdose in a dog.Authors: Connor Ellis, DVMa*, Adesola Odunayo, DVM, MS, DACVECCaa Department of Small Animal Clinical Sciences, University of Florida, Gainesville, FL, USA*Corresponding author: [email protected]: [email protected] of interest: None.No financial support was provided for the preparation of this manuscript.There are no conflicts of interest to declare for this manuscript.Consent statement:Written informed consent was obtained from the owner of this patient to publish this report in accordance with the journal’s patient consent policy.Manuscript type: case reportKeywords: amlodipine, calcium channel blockers, dog, hyperinsulinemiaKey Clinical Message: Hyperinsulinemia-euglycemia therapy (HIET) can be used for treatment of hypotension as well as other clinical signs associated with calcium channel blocker toxicosis. The use of HIET should be considered for amlodipine toxicosis in dogs.IntroductionAmlodipine is a frequently prescribed drug used to control hypertension in human and small animal patients1. This dihydropyridine calcium-channel blocker (CCB) inhibits the influx of calcium ion into the cell through the L-type calcium channel, causing peripheral vasodilation and afterload reduction1. Amlodipine is metabolized by the liver, widely distributed and highly protein bound, with a long half-life of approximately 30 hours in dogs2. While generally well tolerated, reported adverse reactions of amlodipine have been described at doses within the recommended range of 0.1-0.4 mg/kg PO q243–5.The minimum oral toxic dose of each CCB has not been well established in humans or animals, but signs of toxicity have been reported in CCBs at therapeutic doses6. Toxic doses of CCBs have been associated with significant morbidity and mortality6. Clinical signs in affected animals commonly include bradycardia, bradyarrhythmias, and hypotension, with other recognized adverse effects including pleural effusion, gingival hyperplasia, peripheral edema, gastrointestinal distress, hypothermia, central nervous system depression, and electrolyte abnormalities6. While there are currently no consensus guidelines for the treatment of CCB toxicosis in human or veterinary medicine, several retrospective studies and case series have been published in the offering best practice recommendations6–9. A recent veterinary review on CCB toxicosis highlighted decontamination and cardiovascular optimization using fluids and vasoactive medications as the most important interventions. Therapeutic recommendations also included the use of glucagon or intravenous lipid emulsion as secondary options for refractory toxicoses, and other supportive therapies as needed6.An important therapeutic intervention that has resulted in improved patient outcomes over the last 15 years in human patients is hyperinsulinemic-euglycemic therapy (HIET), with multiple publications outlining its efficacy in the human literature10–12. HIET utilizes supraphysiologic doses of insulin, with human doses ranging from 0.5-2.0 U/kg/hour, and concurrent dextrose supplementation to maintain euglycemia12. In brief, blockade of the L-type calcium channels that result in the antihypertensive effect of CCBs also decreases the release of insulin from pancreatic beta cells and reduces glucose uptake by tissues by decreasing sensitivity to insulin, with concurrent vasodilation and hypoperfusion leading to negative inotropy. Increasing insulin availability encourages glucose utilization by the myocardium and suppresses phosphodiesterase III activity, leading to intracellular calcium influx and a positive inotropic effect6. Treatment with HIET is considered a first-line intervention in CCB toxicity in human patients13. There are limited reports of the use of HIET in CCB toxicity in clinical patients in veterinary medicine. There is one case report of survival after HIET was utilized for diltiazem toxicosis in one dog, and another report of HIET therapy utilized for amlodipine toxicosis in a cat, although HIET was discontinued after 3 hours due to refractory hypoglycemia14,15. There are also two animal model studies outlining efficacy of HIET for CCB toxicosis in anesthetized dogs16,17. To the authors’ knowledge, this is the first report describing a protocol for HIET for the treatment of amlodipine toxicosis in the dog.Case descriptionA 1-year-old spayed female mixed breed dog, weighing 14.8 kg, presented to the emergency service for evaluation of ataxia and lethargy approximately 48 hours after ingestion of at least 10 mg of amlodipine (at least a 0.7 mg/kg dose). The ingestion was not witnessed by the owner, but the dog had access to a bottle containing 8 tablets of 10 mg standard release amlodipine, of which one was missing, and half of another tablet had been chewed, resulting in a maximum exposure dose of ~1 mg/kg. The dog reportedly displayed no adverse clinical signs for 24 hours after the potential ingestion, then developed lethargy, anorexia, vomiting, and ataxia in the next 24 hours. After consultation with a poison control center (Pet Poison Helpline, SafetyCall International, LLC), immediate veterinary care was recommended.At presentation, the dog was hypothermic with a rectal temperature of 37.2°C (98.9°F), bradycardic (80 beats/min), tachypneic (52 breaths/min), and dull. The dog was weakly ambulatory and the physical exam was otherwise normal. Systolic blood pressure using a Doppler ultrasonographic device (Parks Medical Electronics, Inc.) was 65 mm Hg. Venous blood was obtained when the peripheral catheter was placed for a PCV/total solids and blood gas with electrolytes. Pertinent findings included a PCV of 54% and a total solids of 66 g/L, a mixed acid-base status, hypocapnia, hyponatremia, hyperkalemia, ionized hypocalcemia, hyperglycemia, hyperlactatemia, decreased bicarbonate, and azotemia (Table 1).TABLE 1. Serial venous blood gas analysis.

Background: Acute Lymphoblastic Leukaemia (ALL) , most common malignancy amongst children with front-line treatment considered major success, 20% of children predicted to either relapse or show resistance to treatment with reported dismal outcomes. Aim: To evaluate clinical characteristics of children diagnosed with refractory / relapsed ALL and to determine 3-year overall survival ouctomes. Method: Retrospective review of patients (aged 1 -14 years) diagnosed with ALL during the period January 2002 to December 2018, data extracted for details of baseline characteristics at diagnosis and at relapse . Results: Total of 347 newly diagnosed children with ALL identified, three induction failures and 28 relapses, with total 31 patients a cohort relapse rate of 9% observed. The male-to-female ratio observed is 4.16:1 and mean duration of CR1 was 26 months : 15 (48%) relapsing ≤ 18 months,seven (23%) during 18 to 36 months and nine (29%) relapsed > 36 months of IF or CR1.Eighteen patients (62%) had isolated BM relapse, six (20%) patients experience isolated Extra-medullary relapse and five (17%) patients experienced BM with other sites. Three-year Overall Survival (OS) of the cohort was 62.3%, while of those patients who achieved CR post first-salvage therapy 3-year OS of 79.5% observed with a statistically significant difference, p value <0.05 comparing to patients who did not achieve remission post first-salvage therapy (3-year OS: 46.4%). The same statistical difference observed in 3-years OS observed comparing duration of remission of CR prior to relapse: ≤ 18 months: 33.2%; 18 – 36 months: 66.7% and > 36 months: 87.5%; the same trend continued when comparing 3-years OS based on risk stratification at relapse: LR: 83.3%; IR: 80% and HR: 44.8%. Conclusion: Incidence and outcomes reported on this study is comparable to internationally reported data with duration of CR1, risk-stratification at relapse and remission status post-salvage therapy determined as significant prognostic factors for survival. No survivial difference amongst patients who received HSCT after induction to those who received chemotherapy, could be attributed to a smaller sample size warranting a multi-institutional observational study. The findings corroborates with the need for novel therapies and treatment approaches in these group of patients

Hodgkin lymphoma is a haematolymphoid malignancy seen in pediatric and adult age groups with an excellent outcome. The current priority of the treatment has been a reduction of acute and chronic morbidities without compromising outcomes. PET-CT scan is an integral component in prognosticating and determining outcomes in Hodgkin lymphoma. Recent studies have highlighted the role of interim PET-CT scan in choosing future therapy. The evidence to support the role of interim PET-CT scan in childhood Hodgkin lymphoma is limited. The aim of the study is to analyse the role of interim PET-CT in the management of childhood HL, and to investigate survival outcomes of children treated at our institute. This is a retrospective study and included all the children with HL treated at our institute in last five years. All the children were treated as per ESMO guidelines. Interim PET-CT scan was done after two cycles of chemotherapy. The correlation of various possible risk factors with the outcomes was examined. Only the interim PET/CT findings were associated with the results. At median follow-up of 34 months, the event free survival (EFS) was 95.7% and overall survival (OS) was 100%. Interim PET-CT plays an important role in treatment modification without compromising on the outcomes in children with HL. Survival rates were consistent with those reported in published literature.

Understanding how ecological and environmental changes, anthropogenic activities, and climate have driven and will direct animals’ development and predicting their prospective distribution profiles in the Quaternary are essential to making a tangible conservation strategy. Macaques (Macaca) distributed in mainland East Asia provide an ideal research model for such an effort. We reconstruct macaques’ geographic distribution profiles during the Quaternary, from the last inter-glaciation (LIG, 120,000 - 140,000 years BP), the Last Glacial Maximum (LGM, 22,000 years BP), and the present (1970-2000) – based on which we deduce their perspective distribution in the 2050s. The results show their suitable habitats during LIG and LGM were mainly in Southwest, Central, and Coastal China. A noticeable distribution reduction started in LIG and persisted until the current (1970-2000). Their distribution centroid would shift northward to mountainous regions, mainly in Southwest China, where more migration corridors would be reserved for their future development. Also, the results indicate that China’s Protected Area currently does not cover more than 87% of macaques’ habitats, a dismal situation for their conservation. Finally, this study proclaims that the conservation priority of the macaques in the years to come should focus on Southwest China – their future refuge region in Quaternary.

Ntšhe, a Setswana name for sweet reeds, a sweet sorghum variety ( Sorghum bicolor (L)) is a commercial crop consumed as a delicacy in enormous quantities in Botswana and in southern Africa at large. Pre and post-harvest losses due to infestation by larval stages of stem borer moths, Chilo partellus often lead to severe financial losses as consumers condemn the worm-infested sweet reeds as unpalatable, “ Mo gase ntšhe tota tota” (this is not a sweet reed). Valorisation of condemned sweet reeds is one attractive route to reduce economic losses. Here, we took advantage of our understanding of yeast-insect interactions to isolate yeasts associated with larval stages of the stem borer moths and investigated their potential for production of an alcoholic sweet reed beverage. We isolated 33 yeast strains representing from the galleries and frass as well as from the guts of the larval moths. Assessment of their ability to ferment the simplest sugar, glucose, resulted in identification of 14 strains belonging to Hanseniaspora and Candida genera. These strains were further assessed for their capacity to ferment by calculating the rate of accumulation of carbon dioxide and ethanol when grown in the principal sugars found in sweet sorghum juice as sole carbon sources as well in sweet sorghum juice. In addition, as an industrially relevant trait, we tested the potential of the strains to tolerate brewing/fermentation-associated stresses. Furthermore, we assessed the aromatic complexity of the produced beverage. Our results suggest that non-conventional yeasts associated with the larval moths have potential for valorization of condemned sweet sorghum stalks to produce a sweet sorghum beverage.

A document by Rafat Noeiaghdam. Click on the document to view its contents.

Current methodologies of genome-wide Single Nucleotide Polymorphism (SNP) genotyping produce large amounts of missing data that may affect statistical inference and bias the outcome of experiments. Genotype imputation is routinely used in well-studied species to buffer the impact in downstream analysis and several algorithms are available to fill in missing genotypes. The lack of reference haplotype panels precludes the use of these methods in genomic studies on non-model organisms. As an alternative, machine learning algorithms are employed to explore the genotype data and to estimate the missing genotypes. Here, we propose an imputation method based on Self-Organizing Maps (SOM), a widely used neural networks formed by spatially distributed neurons that cluster similar inputs into close neurons. We follow a classical approach that explores genotype datasets to select SNP loci for each query missing SNP genotype to build training sets, and that initializes and trains the neural networks to finally use the SOM-derived clustering to impute the best genotype. To automate the imputation process, we have implemented GTIMPUTATION, an open source application programmed in Python3 and with a user-friendly GUI to facilitate the whole process. The method performance was validated by comparing its accuracy, precision and sensitivity on several benchmark genotype datasets with other available imputation algorithms. Our approach produced highly accurate and precise genotype imputations and outperformed other algorithms, especially for datasets from mixed populations with unrelated individuals.