Search results for: acute leukemia

Authors: Aihong Zhao, Priya Sastry, Mark L Field, Mohamad Bashir, Arvind Singh, David Richens

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Objectives: Intramural haematoma (IMH) is one of the pathologies, along with acute aortic dissection, that present as Acute Aortic Syndrome (AAS). Evidence suggests that unlike aortic dissection, some intramural haematomas may regress with medical management. However, intramural haematomas have been traditionally managed like acute aortic dissections. Given that some of these pathologies may regress with conservative management, it would be useful to be able to identify which of these may not need high risk emergency intervention. A computational aortic model was used in this study to try and identify intramural haematomas with risk of progression to aortic dissection. Methods: We created a computational model of the aorta with luminal blood flow. Reports in the literature have identified 11 mm as the radial clot thickness that is associated with heightened risk of progression of intramural haematoma. Accordingly, haematomas of varying sizes were implanted in the modeled aortic wall to test this hypothesis. The model was exposed to physiological blood flows and the stresses and strains in each layer of the aortic wall were recorded. Results: Size and shape of clot were seen to affect the magnitude of aortic stresses. The greatest stresses and strains were recorded in the intima of the model. When the haematoma exceeded 10 mm in all dimensions, the stress on the intima reached breaking point. Conclusion: Intramural clot size appears to be a contributory factor affecting aortic wall stress. Our computer simulation corroborates clinical evidence in the literature proposing that IMH diameter greater than 11 mm may be predictive of progression. This preliminary report suggests finite element modelling of the aortic wall may be a useful process by which to examine putative variables important in predicting progression or regression of intramural haematoma.

Keywords: intramural haematoma, acute aortic syndrome, finite element analysis,

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Authors: M. Hakami, A. Jammaly, I. Attafi, M. Oraiby, M. Jeraiby

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We reviewed an unusual case of a 65-year-old male taking an herbal mixture containing compounds with anticholinesterase activity for a long period of time, presented with acute my myocardial infarction and multiple organ dysfunction syndrome followed by death. Clinically, there are findings correlated with anticholinesterase activity, such as bilateral miosis, diaphoresis, vomiting and fasciculation without a history of any toxic ingestion or exposure. Gas chromatography–mass spectrometry screening studies identified the presence of thymol, anethole in the herbal extract and butylated hydroxytoluene in the blood sample. Hence, with this case report, we intend to highlight the necessity of evaluating the long-term use of the herbal mixture.

Keywords: cholinesterase inhibitors, thymole, anethole, butylatedhydroxytoluene, cardiac toxicity, myocardial infarction

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Authors: Maha Salah Abdullah Ismail, Mahmoud M. Alsagheir, Mohammed Salah Abd Allah

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Acute respiratory distress syndrome (ARDS) is characterized by permeability pulmonary edema and refractory hypoxemia. Lung-protective ventilation is still the key of better outcome in ARDS. Prone position reduces the trans-pulmonary pressure gradient, recruiting collapsed regions of the lung without increasing airway pressure or hyperinflation. Prone ventilation showed improved oxygenation and improved outcomes in severe hypoxemic patients with ARDS. This study evaluates the effect of prone positioning on mechanically ventilated patients with ARDS. A quasi-experimental design was carried out at Critical Care Units, on 60 patients. Two tools were utilized to collect data; Socio demographic, medical and clinical outcomes data sheet. Results of the present study indicated that prone position improves oxygenation in patients with severe respiratory distress syndrome. The study recommended that use prone position in patients with severe ARDS, as early as possible and for long sessions. Also, replication of this study on larger probability sample at the different geographical location is highly recommended.

Keywords: acute respiratory distress syndrome, critical care, mechanical ventilation, prone position

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Authors: Kin On Kwok, Vivian Wei, Benjamin Cowling, Steven Riley, Jonathan Read

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Background: The role of social contact behavior measured in different contact constructs in the transmission of respiratory pathogens with acute respiratory illness (ARI) remains unclear. We, therefore, aim to depict the individual pattern of ARI in the community and investigate the association between different contact dimensions and ARI in Hong Kong. Methods: Between June 2013 and September 2013, 620 subjects participated in the last two waves of recruitment of the population based longitudinal phone social contact survey. Some of the subjects in this study are from the same household. They are also provided with the symptom diaries to self-report any acute respiratory illness related symptoms between the two days of phone recruitment. Data from 491 individuals who were not infected on the day of phone recruitment and returned the symptom diaries after the last phone recruitment were used for analysis. Results: After adjusting different follow-up periods among individuals, the overall incidence rate of ARI was 1.77 per 100 person-weeks. Over 75% ARI episodes involve running nose, cough, sore throat, which are followed by headache (55%), malagia (35%) and fever (18%). Using a generalized estimating equation framework accounting for the cluster effect of subjects living in the same household, we showed that both daily number of locations visited with contacts and the number of contacts, explained the ARI incidence rate better than only one single contact construct. Conclusion: Our result suggests that it is the intertwining property of contact quantity (number of contacts) and contact intensity (ratio of subject-to-contact) that governs the infection risk by a collective set of respiratory pathogens. Our results provide empirical evidence that multiple contact constructs should be incorporated in the mathematical transmission models to feature a more realistic dynamics of respiratory disease.

Keywords: acute respiratory illness, longitudinal study, social contact, symptom diaries

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Authors: G. E. Forcados, M. L. Shu, C. N. Chinyere

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Acalypha wilkesiana is a medicinal plant commonly used in most parts of West Africa as a decoction in treating several human diseases. Existing literature on its toxicity is predominantly on the organic extracts in contrast to the routine use of hot aqueous extracts as decoction. The aim of this study was to examine the phytochemical profile and sub-acute toxicity of A. wilkesiana leaf extracts in albino rats. Three groups of 8 experimental rats each were administered 300 mg/kg, 600 mg/kg and 1200 mg/kg body weight per day for 14 days while a fourth (control) group took tap-water. On day 15, the rats were sacrificed, and blood collected. Biochemical and hematological parameters were analysed and histopathological examination of liver and kidney were performed. There was significant increase (p<0.05) in the levels of some biochemical parameters (AST, ALT, creatinine, urea) in all the test groups compared to control. Histopathological examination of the liver revealed centrilobular degeneration and necrosis with sinusoidal dilatation as well as polymorphonuclear and mononuclear infiltration, likewise severe glomerular and tubular degeneration and necrosis with hemorrhage in the kidney at all dose levels. The results from this study suggest that aqueous leaf extract of A. wilkesiana is hepatotoxic and nephrotoxic at dose levels of 300 mg/kg and above. Therefore, precautionary measures are necessary for home use of the leaf extract of A. wilkesiana.

Keywords: acute toxicity, A. wilkesiana, aqeous extract, albino rats, biochemical and haematological parameters, histopathological examination

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Authors: Njeim Pressila, Hajj-Boutros Guy, Antony D. Karelis

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Background: Acute resistance exercise is associated with an elevated inflammation response, which could lead to delayed onset muscle soreness (DOMS). There is evidence that suggests that ingesting foods that have anti-inflammation properties may help reduce DOMS. A vegan diet has also been shown to be an anti-inflammatory diet which could, in turn, decrease DOMS. Objective and hypothesis: The purpose of the present study will be to compare markers of DOMS between vegans and omnivores after acute resistance exercise in young females. We hypothesize that vegans will have a better recovery of DOMS markers after a resistance exercise session compared to omnivores. Methods: Population: We will recruit30 vegans and 30 omnivores to participate in this study. Allvolunteers will follow either a vegan or an omnivore diet for at least 2 years. Anthropometric measurements, body composition, musclestrength (leg and chest press), markers of DOMS (swelling, pain, and stiffness), and dietary factors, as well as a wellness and anxiety questionnaire will be measured. All participants will also perform an acute resistance exercise session in order to induce DOMS. Pertinence: This project will give us a better understanding on the recovery process of vegans after a resistance training session and, as such, provide useful information to health professionals and athletes/coaches (kinesiologists and nutritionists)

Keywords: vgeans, omnivores, delayed onset muscle soreness, pain, stifness

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Authors: Abdul-Ganiyu Fuseini, Bernice Redley, Helen Rawson, Lenore Lay, Debra Kerr

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Aim: The study aimed to develop and validate a culturally appropriate patient-reported outcome measure for measuring dignity for older adults during acute hospital admissions. Design: A three-phased mixed-method sequential exploratory design was used. Methods: Concept elicitation and generation of items for the scale was informed by older adults’ perspectives about dignity during acute hospitalization and a literature review. Content validity evaluation and pre-testing were undertaken using standard instrument development techniques. A cross-sectional survey design was conducted involving 270 hospitalized older adults for evaluation of construct and convergent validity, internal consistency reliability, and test–retest reliability of the scale. Analysis was performed using Statistical Package for the Social Sciences, version 25. Reporting of the study was guided by the Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) checklist. Results: We established the 15-item Hospitalized Older Adults’ Dignity Scale that has a 5-factor structure: Shared Decision-Making (3 items); Healthcare Professional-Patient Communication (3 items); Patient Autonomy (4 items); Patient Privacy (2 items); and Respectful Care (3 items). Excellent content validity, adequate construct and convergent validity, acceptable internal consistency reliability, and good test-retest reliability were demonstrated. Conclusion: We established the Hospitalized Older Adults Dignity Scale as a valid and reliable scale to measure dignity for older adults during acute hospital admissions. Future studies using confirmatory factor analysis are needed to corroborate the dimensionality of the factor structure and external validity of the scale. Routine use of the scale may provide information that informs the development of strategies to improve dignity-related care in the future. Impact: The development and validation of the Hospitalized Older Adults Dignity Scale will provide healthcare professionals with a feasible and reliable scale for measuring older adults’ dignity during acute hospitalization. Routine use of the scale may enable the capturing and incorporation of older patients’ perspectives about their healthcare experience and provide information that informs the development of strategies to improve dignity-related care in the future.

Keywords: dignity, older adults, hospitalisation, scale, patients, dignified care, acute care

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Authors: C. Scurr, J. Jeans, S. Srivastava

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Introduction: Acute type two respiratory failure (T2RF) has become a common presentation over the last two decades primarily due to an increase in the prevalence of chronic lung disease. Acute exacerbations can be managed either medically or in combination with non-invasive ventilation (NIV) which should be monitored with regular arterial blood gas samples (ABG). Arterial lines allow more frequent arterial blood sampling with less patient discomfort. We present the experience from a teaching hospital emergency department (ED) and level 2 medical high-dependency unit (HDU) that together form the pathway for management of acute type 2 respiratory failure. Methods: Patients acutely presenting to Charing Cross Hospital, London, with T2RF requiring non-invasive ventilation (NIV) over 14 months (2011 to 2012) were identified from clinical coding. Retrospective data collection included: demographics, co-morbidities, blood gas numbers and timing, if arterial lines were used and who performed this. Analysis was undertaken using Microsoft Excel. Results: Coding identified 107 possible patients. 69 notes were available, of which 41 required NIV for type 2 respiratory failure. 53.6% of patients had an arterial line inserted. Patients with arterial lines had 22.4 ABG in total on average compared to 8.2 for those without. These patients had a similar average time to normalizing pH of (23.7 with arterial line vs 25.6 hours without), and no statistically significant difference in mortality. Arterial lines were inserted by Foundation year doctors, Core trainees, Medical registrars as well as the ICU registrar. 63% of these were performed by the medical registrar rather than ICU, ED or a junior doctor. This is reflected in that the average time until an arterial line was inserted was 462 minutes. The average number of ABGs taken before an arterial line was 2 with a range of 0 – 6. The average number of gases taken if no arterial line was ever used was 7.79 (range of 2-34) – on average 4 times as many arterial punctures for each patient. Discussion: Arterial line use was associated with more frequent arterial blood sampling during each inpatient admission. Additionally, patients with an arterial line have less individual arterial punctures in total and this is likely more comfortable for the patient. Arterial lines are normally sited by medical registrars, however this is normally after some delay. ED clinicians could improve patient comfort and monitoring thus allowing faster titration of NIV if arteral lines were regularly inserted in the ED. We recommend that ED doctors insert arterial lines when indicated in order improve the patient experience and facilitate medical management.

Keywords: non invasive ventilation, arterial blood gas, acute type, arterial line

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Authors: Noura El-Ahmady El-Naggar

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Among the antitumour drugs, bacterial enzyme L-asparaginase has been employed as the most effective chemotherapeutic agent in pediatric oncotherapy especially for acute lymphoblastic leukemia. Glutaminase free L-asparaginase producing actinomycetes were isolated from soil samples collected from Egypt. Among them, a potential culture, strain NEAE-119, was selected and identified on the basis of morphological, cultural, physiological and biochemical properties, together with 16S rDNA sequence as Streptomyces olivaceus NEAE-119 and sequencing product(1509 bp) was deposited in the GenBank database under accession number KJ200342. The optimization of different process parameters for L-asparaginase production by Streptomyces olivaceus NEAE-119 using Plackett–Burman experimental design and response surface methodology was carried out. Fifteen nutritional variables (temperature, pH, incubation time, inoculum size, inoculum age, agitation speed, dextrose, starch, L-asparagine, KNO3, yeast extract, K2HPO4, MgSO4.7H2O, NaCl and FeSO4. 7H2O) were screened using Plackett–Burman experimental design. The most positive significant independent variables affecting enzyme production (temperature, inoculum age and agitation speed) were further optimized by the central composite face-centered design -response surface methodology. As a result, a medium of the following formula is the optimum for producing an extracellular L-asparaginase in the culture filtrate of Streptomyces olivaceus NEAE-119: Dextrose 3g, starch 20g, L-asparagine 10g, KNO3 1g, K2HPO4 1g, MgSO4.7H2O 0.1g, NaCl 0.1g, pH 7, temperature 37°C, agitation speed 200 rpm/min, inoculum size 4%, v/v, inoculum age 72 h and fermentation period 5 days.

Keywords: Streptomyces olivaceus NEAE-119, glutaminase free L-asparaginase, production, Plackett-Burman design, central composite face-centered design, 16S rRNA, scanning electron microscope

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Authors: Hyuk-Hoon Kim, Young Gi Min

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Background: In carbon monoxide (CO) intoxication, the pathophysiology of delayed neurological sequelae (DNS) is very complex and remains poorly understood. And predicting whether patients who exhibit resolved acute symptoms have escaped or will experience DNS represents a very important clinical issue. Brain magnetic resonance (MR) imaging has been conducted to assess the severity of brain damage as an objective method to predict prognosis. And co-ingestion of a second poison in patients with intentional CO poisoning occurs in almost one-half of patients. Among patients with co-ingestions, 66% ingested ethanol. We assessed the effects of ethanol on neurologic sequelae prevalence in acute CO intoxication by means of abnormal lesion in brain MR. Method: This study was conducted retrospectively by collecting data for patients who visited an emergency medical center during a period of 5 years. The enrollment criteria were diagnosis of acute CO poisoning and the measurement of the serum ethanol level and history of taking a brain MR during admission period. Official readout data by radiologist are used to decide whether abnormal lesion is existed or not. The enrolled patients were divided into two groups: patients with abnormal lesion and without abnormal lesion in Brain MR. A standardized extraction using medical record was performed; Mann Whitney U test and logistic regression analysis were performed. Result: A total of 112 patients were enrolled, and 68 patients presented abnormal brain lesion on MR. The abnormal brain lesion group had lower serum ethanol level (mean, 20.14 vs 46.71 mg/dL) (p-value<0.001). In addition, univariate logistic regression analysis showed the serum ethanol level (OR, 0.99; 95% CI, 0.98 -1.00) was independently associated with the development of abnormal lesion in brain MR. Conclusion: Ethanol could have neuroprotective effect in acute CO intoxication by sedative effect in stressful situation and mitigative effect in neuro-inflammatory reaction.

Keywords: carbon monoxide, delayed neuropsychological sequelae, ethanol, intoxication, magnetic resonance

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Authors: Rachael Collins, George Lafford

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We report a case of a generally fit and a well 54-year-old gentleman who presented with a two-day history of worsening left-sided otorrhea, headache, neck stiffness, vomiting and pyrexia on the background of a seven-week history of OE. His condition progressed dramatically as he developed symptoms consistent with acute complete cervical cord syndrome with radiological evidence of skull base osteomyelitis, parapharyngeal, retropharyngeal and paravertebral abscesses and sigmoid sinus thrombus. Ultimately he made a significant, although not complete, recovery. This case is unique in demonstrating how OE can develop into a potentially life-threatening condition. It emphasizes the importance of early diagnosis and treatment of OE, the recognition of ‘red flag’ symptoms and highlights the importance of a multi-disciplinary team (MDT) approach when managing complex complications of OE.

Keywords: ENT, neurology, otology, MDT

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Authors: Wahyu Purnama Putra, Artono Isharanto

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Thoracic trauma is trauma that hits the thoracic wall or intrathoracic organs, either due to blunt trauma or sharp trauma. Thoracic trauma often causes impaired ventilation-perfusion due to damage to the lung parenchyma. This results in impaired tissue oxygenation, which is one of the causes of acute respiratory distress syndrome (ARDS). These changes are caused by the release of pro-inflammatory mediators, plasmatic proteins, and proteases into the alveolar space associated with ongoing edema, as well as oxidative products that ultimately result in severe inhibition of the surfactant system. This study aims to predict the incidence of acute respiratory distress syndrome (ARDS) through human neutrophil elastase levels. This study examines the relationship between plasma elastase levels as a predictor of the incidence of ARDS in thoracic trauma patients in Malang. This study is an observational cohort study. Data analysis uses the Pearson correlation test and ROC curve (receiver operating characteristic curve). It can be concluded that there is a significant (p= 0.000, r= -0.988) relationship between elastase levels and BGA-3. If the value of elastase levels is limited to 23.79 ± 3.95, the patient will experience mild ARDS. While if the value of elastase levels is limited to 57.68 ± 18.55, in the future, the patient will experience moderate ARDS. Meanwhile, if the elastase level is between 107.85 ± 5.04, the patient will likely experience severe ARDS. Neutrophil elastase levels correlate with the degree of severity of ARDS incidence.

Keywords: ARDS, human neutrophil elastase, severity, thoracic trauma

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Authors: Ghada Hasabo, Mahmoud Saber Elbasiouny, Mervat Abdelsalam, Sherin Ghaleb, Niveen Eldessouky

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In the near future, nanotechnology is envisaged for large scale use. Hence health and safety issues of nanoparticles should be promptly addressed. In the present study the acute hepatoxicity assessment due to high single oral dose of nano iron and micro iron particles were studied. The normal daily activities, biochemical alterations, blood coagulation, histopathological changes in Wister rats were the aspect of the toxicological assessment.This work found that significant alterations in biochemical enzymes (serum iron level, liver enzymes, albumin, and bilirubin levels), blood coagulation (PT, PC, INR), and histopathological changes occurred more prominently in the nano iron particle treated group.

Keywords: nanobiotechnology, nanosystems, nanomaterials, nanotechnology

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Authors: Patti Kastanias, Wei Wang, Karyn Mackenzie, Sandra Robinson, Susan Wnuk

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Obesity is a worldwide epidemic and is recognized as a chronic disease. Pain in the obese individual is a multidimensional issue. An increase in BMI is positively correlated with pain incidence and severity, especially in central obesity where individuals are twice as likely to have chronic pain. Both obesity and chronic pain are also associated with mood disorders. Pain is worse among obese individuals with depression and anxiety. Bariatric surgery provides patients with an effective solution for long-term weight loss and associated health problems. However, not much is known about acute and chronic pain after bariatric surgery and its contributing factors, including mood disorders. Nurse practitioners (NPs) at one large multidisciplinary bariatric surgery centre led two studies to examine acute and chronic pain and pain management over time after bariatric surgery. The purpose of the initial study was to examine the incidence and severity of acute and chronic pain after bariatric surgery. The aim of the secondary study was to further examine chronic pain, specifically looking at psychological factors that influence severity or incidence of both neuropathic and somatic pain as well as changes in opioid use. The initial study was a prospective, longitudinal study where patients having bariatric surgery at one surgical center were followed up to 6 months postop. Data was collected at 7 time points using validated instruments for pain severity, pain interference, and patient satisfaction. In the second study, subjects were followed longitudinally starting preoperatively and then at 6 months and 1 year postoperatively to capture changes in chronic pain and influencing variables over time. Valid and reliable instruments were utilized for all major study outcomes. In the first study, there was a trend towards decreased acute post-operative pain over time. The incidence and severity of chronic pain was found to be significantly reduced at 6 months post bariatric surgery. Interestingly, interference of chronic pain in daily life such as normal work, mood, and walking ability was significantly improved at 6 months postop however; this was not the case with sleep. Preliminary results of the secondary study indicate that pain severity, pain interference, anxiety and depression are significantly improved at 6 months postoperatively. In addition, preoperative anxiety, depression and emotional regulation were predictive of pain interference, but not pain severity. The results of our regression analyses provide evidence for the impact of pre-existing psychological factors on pain, particularly anxiety in obese populations.

Keywords: bariatric surgery, mood disorders, obesity, pain

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Authors: Yu-Tzu Hsu, Alice Wu, Hsiang-Kuang Tseng

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Introduction: Hepatitis A virus causes acute hepatitis and is usually transmitted by a fecal-oral route of food contamination, which is more prevalent in areas with poor hygienic practices. However, we described a hepatitis A outbreak associated with a fecal-oral route of sexual behavior in men who has sex with men (MSM) in Northern Taiwan. Methods: We retrospectively collected patients with acute HAV infection in MacKay Memorial Hospital, Taipei, Taiwan between July 2015 and November 2016. Demographic data (age, gender, onset time and infection risk), laboratory data (GOT, GPT, bilirubin, HIV status, HBsAg, HCV antibody and syphilis), clinical symptoms and travel history with a foreign tour were analyzed. We compared variables between HIV and non-HIV group. Unless otherwise stated, continuous variables were expressed as mean ± SD, and categorical variables were expressed as number (percentage) for each item. The t test for continuous variables was applied for the comparison between two groups and chi-square for categorical variables were applied for measures of association. Results: We collected 80 cases during the study period. Among them, 54 (67.5%) cases were MSM and 43 (53.8%) cases were HIV positive. The average age was 32.6±7.59 years-old. The average value of initial liver function was 1324 IU/L for AST (GOT), 2100 IU/L for ALT (GPT), and 5.82 mg/dL for bilirubin. We found seven (8.6%) cases were in the status of HBV carrier, five (6.3%) cases were positive for HCV antibody, and 15 (18.6%) cases were co-infected with syphilis. With regards to associated symptoms, 32 (40%) had fever, 46 (57.5%) had nausea, 34 (42.5%) had abdominal discomfort and 46 (57.5%) had general malaise. To compare the non-HIV patients with HIV patients, HIV patients were more likely to be male (p=0.008), MSM (p=0.000), co-infected syphilis (p=0.000) and slowly improving liver function of transaminases (p=0.033, 0.027). Conclusion: The HAV outbreak in Northern Taiwan was mainly occurred in MSM population. Hereafter, our cohort data support a policy in Taiwan to provide one dose of free HAV vaccine shot in this population. Hopefully, the outbreak could be stop by the free vaccine policy and public education.

Keywords: acute hepatitis A, men who has sex with men, human immunodeficiency virus, vaccine

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Authors: D. M. Lewis, L. Frisby, U. Stead

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Objective: Many patients are receiving invasive treatments in acute care or are dying in hospital without having had comprehensive goals of care conversations. Some of these treatments may not align with the patient’s wishes, may be futile, and may cause unnecessary suffering. While many staff may recognize the benefits of engaging patients and families in Serious Illness Conversations (a goal of care framework developed by Ariadne Labs in Boston), few staff feel confident and/or competent in having these conversations in acute care. Another barrier to having these conversations may be due to a lack of incorporation in the current workflow. An educational exercise, titled the Two Question Challenge, was initiated on four medical units across two Vancouver Coastal Health (VCH) hospitals in attempt to engage the entire interdisciplinary team in asking patients and families questions around goals of care and to improve the documentation of these expressed wishes and preferences. Methods: Four acute care units across two separate hospitals participated in the Two Question Challenge. On each unit, over the course of two eight-hour shifts, all members of the interdisciplinary team were asked to select at least two questions from a selection of nine goals of care questions. They were asked to pose these questions of a patient or family member throughout their shift and then asked to document their conversations in a centralized Advance Care Planning/Goals of Care discussion record in the patient’s chart. A visual representation of conversation outcomes was created to demonstrate to staff and patients the breadth of conversations that took place throughout the challenge. Staff and patients were interviewed about their experiences throughout the challenge. Two palliative approach leads remained present on the units throughout the challenge to support, guide, or role model these conversations. Results: Across four acute care medical units, 47 interdisciplinary staff participated in the Two Question Challenge, including nursing, allied health, and a physician. A total of 88 questions were asked of patients, or their families around goals of care and 50 newly documented goals of care conversations were charted. Two code statuses were changed as a result of the conversations. Patients voiced an appreciation for these conversations and staff were able to successfully incorporate these questions into their daily care. Conclusion: The Two Question Challenge proved to be an effective way of having teams explore the goals of care of patients and families in an acute care setting. Staff felt that they gained confidence and competence. Both staff and patients found these conversations to be meaningful and impactful and felt they were notably different from their usual interactions. Documentation of these conversations in a centralized location that is easily accessible to all care providers increased significantly. Application of the Two Question Challenge in non-medical units or other care settings, such as long-term care facilities or community health units, should be explored in the future.

Keywords: advance care planning, goals of care, interdisciplinary, palliative approach, serious illness conversations

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Authors: Adar Pelah, Avraham Adelman, Amanda Balash, Jake Mitchell, Mattan J. Pelah, Viswadeep Sarangi, Xin Chen Cai, Zadok Storkey, Gregg B. Fields, Ximena Levy, Ali A. Danesh

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Introduction: Post-Acute Sequelae of Severe Acute Respiratory Syndrome Coronavirus 2 infection (PASC), or Long COVID, while primarily a respiratory disorder, can also include dizziness, lasting weeks to months in individuals who had previously tested positive for COVID-19. This study utilized gait analysis to assess the potential vestibular effects of PASC on the presentation of gait anomalies. Materials and Methods: The study included 11 participants who tested positive for COVID-19, a mean of 2.8 months prior to gait testing (PP=11), and 8 control participants who did not test positive for COVID-19 (NP=8). Participants walked 7.5m at three self-selected speeds: ‘slow,’ ‘normal,’ and ‘fast.’ Mean walking speeds were determined for each speed and overall range from four laps on an instrumented walkway using video capture. Results: A Z-test at 0.05 significance was used for speed range, ‘normal’ and ‘fast’ at the lower tail, and for ‘slow’ at the higher tail. Average speeds (m/s) were: ‘slow’ (PP=0.709, NP=0.678), ‘normal’ (PP=1.141, NP=1.170), ‘fast’ (PP=1.529, NP=1.821), average range (PP=0.846, NP=1.143). Significant speed decreases between PP and NP were observed in ‘fast’ (-17.43%) and average range (-29.86%), while changes in ‘slow’ (+2.44%) and ‘normal’ (-4.39%) speeds were not significant. Conclusions: Long COVID is a recognized disability (Americans with Disabilities Act), and although it presents variably, dizziness, vertigo, and tinnitus are not uncommon in COVID-19 infection. These results suggest that potential inner-ear damage may persist and manifest in gait changes even after recovery from acute illness. Further research with a larger sample size may indicate the need for providers to consider PASC when diagnosing patients with vestibular dysfunction.

Keywords: gait analysis, long-COVID, vestibular dysfunction, walking speed

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Authors: Irfan Khan, Chiemeka David Ekene Arize, Hilly Swami, Suprabha Jha

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Background: The rise of myocardial infarction (MI) among young adults, especially those with psychiatric conditions on antipsychotic medications, highlights the need to explore non-traditional cardiovascular risk factors. Case Presentation: We discuss a 24-year-old male with acute MI, diagnosed with an acute anxiety disorder, treated with risperidone and quetiapine, and with a history of occasional smoking. Despite no significant medical history, his presentation underscores the complex interactions between psychiatric conditions, antipsychotic medication, and lifestyle choices in the etiology of MI. Discussion: This case sheds light on the intricate relationship between minimal smoking habits, the use of atypical antipsychotics, and psychiatric illness as contributory factors to cardiovascular risk in young patients. It suggests a synergistic effect, amplifying the risk of MI, which is not adequately captured by traditional risk models. Conclusion: The case emphasizes the importance of an integrated care appro ach for young MI patients with psychiatric conditions and highlights the urgent need for further research to understand the compounded cardiovascular risk posed by psychiatric medications and lifestyle factors. It advocates for comprehensive risk assessments that consider these non-traditional factors to improve outcomes for this vulnerable patient population.

Keywords: myocardial infarction, young adults, psychiatric illness, antipsychotic medications, smoking

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Authors: Ghada A. Gamea, Nabila A. Yaseen, Ahmed A. Othman, Ahmed S. Tawfik

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Infection with Toxoplasma gondii can cause serious complications in pregnant women, leading to abortion, stillbirth, and congenital anomalies in the fetus. Definitive diagnosis of T. gondii acute infection is therefore critical for the clinical management of a mother and her fetus. This study was conducted on 250 pregnant females in the first trimester who were inpatients or outpatients at Obstetrics and Gynaecology Department at Tanta University Hospital. Screening of the selected females was done for the detection of immunoglobulin (IgG and IgM), and all subjects were submitted to history taking through a questionnaire including personal data, risk factors for Toxoplasma, complaint and history of the present illness. Thirty-eight samples, including 18 IgM +ve and 20 IgM-ve cases were further investigated by the avidity and IgE ELISA tests. The seroprevalence of toxoplasmosis in pregnant women was (42.8%) based on the presence of IgG antibodies in their sera. Contact with cats and consumption of raw or undercooked meat are important risk factors that were associated with toxoplasmosis in pregnant women. By serology, it could be observed that in the IgM +ve group, only one case (5.6%) showed an acute pattern by using the avidity test, though 10 (55.6%) cases were found to be acute by the IgE assay. On the other hand, in the IgM –ve group, 3 (15%) showed low avidity, but none of them was positive by using the IgE assay. In conclusion, there is no single serological test that can be used to confirm whether T. gondii infection is recent or was acquired in the distant past. A panel of tests for detection of toxoplasmosis will certainly have higher discriminatory power than any test alone.

Keywords: diagnosis, serology, seroprevalence, toxoplasmosis

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Authors: Grzegorz Swiderski, Agata Jablonska-Trypuc, Natalia Popow, Renata Swislocka, Wlodzimierz Lewandowski

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Doxorubicin belongs to the group of anthracycline antitumor antibiotics. Because of the wide spectrum of actions, it is one of the most widely used anthracycline antibiotics, including the treatment of breast, ovary, bladder, lung cancers as well as neuroblastoma, lymphoma, leukemia and myeloid leukemia. Antitumor activity of doxorubicin is based on the same mechanisms as for most anthracyclines. Like the metal ions affect the nucleic acids on many biological processes, so the environment of the metal chelates of antibiotics can have a significant effect on the pharmacological properties of drugs. Complexation of anthracyclines with metal ions may contribute to the production of less toxic compounds. In the framework of this study, the composition of complexes obtained in aqueous solutions of doxorubicin with metal ions (Cu2+ and Zn2+). Complexation was analyzed by spectrophotometric titration in aqueous solution at pH 7.0. The pH was adjusted with 0.02M Tris-HCl buffer. The composition of the complexes found was Cu: doxorubicin (1: 2) and a Zn: doxorubicin (1: 1). The effect of Dox, Dox-Cu and Dox-Zn was examined in MCF-7 breast cancer cell line, which were obtained from American Type Culture Collection (ATCC). The compounds were added to the cultured cells for a final concentration in the range of 0,01µM to 0,5µM. The number of MCF-7 cells with division into living and dead, was determined by direct counts of cells with the use of trypan blue dye using LUNA Logos Biosystems cell counter. ApoTox-Glo Triplex Assay (Promega, Madison, Wisconsin, USA) was used according to the manufacturer’s instructions to measure the MCF-7 cells’ viability, cytotoxicity and apoptosis. We observed a decrease in cells proliferation in a dose-dependent manner. An increase in cytotoxicity and decrease in viability in the ApoTox Triplex assay was also showed for all tested compounds. Apoptosis, showed as caspase 3/7 activation, was observed only in Dox treatment. In Dox-Zn and Dox-Cu caspase 3/7 activation was not observed. This work was financially supported by National Science Centre, Poland, under the research project number 2014/13/B/NZ7/02 352.

Keywords: anticancer properties, anthracycline antibiotic, doxorubicine, metal complexes

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Authors: Angelis P. Barlampas

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PURPOSE: The purpose of this paper is to demonstrate the important role of taking an appropriate and detailed history, in order to reach the best possible diagnostic conclusion. MATERIAL: A patient presented to the emergency department due to the sudden onset of continuous abdominal pain, during the last hour and with the clinical symptoms of an acute abdomen. During the clinical examination, signs of peritoneal irritation and diffuse abdominal tenderness were found. The rest of the clinical and laboratory tests did not reveal anything important. From the reported medical history, nothing of note was found, except for the report of a large liver cyst, for which he was advised not to take any further action, except from regular ultrasound examination . METHOD: A computed tomography examination was performed after per os administration of gastrografin, which revealed a hyperdense ascitic effusion, similar in density to that of gastrografin within the intestinal tract. The presence of a large cyst of the left hepatic lobe was confirmed, contacting and pushing against the stomach. In the area of the contact between the liver cyst and the pylorus, there were extraluminal air bubbles and local opacity of the peritoneal fat, with a small hyperdense effusion. Result : The above, as well as the absence of a history of stomach ulcer or recent trauma, or other pathology, argue in favor of acute pyloric perforation, due to mural necrosis, in response to chronic external pressure from the pre-existing large liver cyst.

Keywords: perforation, stomach, large liver cyst, CT abdomen, acute abdominal pain, intraperitoneal leakage, constrast leakage

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Authors: Shweh Fern Loo, Jun Yin Ong, Than Zaw Oo

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Acute respiratory distress syndrome (ARDS) is a common serious condition of bilateral lung infiltrates that develops secondary to various underlying conditions such as diseases or injuries. ARDS with severe hypercapnia is associated with higher ICU mortality and morbidity. Venovenous Extracorporeal membrane oxygenation (VV-ECMO) support has been established to avert life-threatening hypoxemia and hypercapnic respiratory failure despite optimal conventional mechanical ventilation. However, VV-ECMO is relatively not advisable in particular groups of patients, especially in multi-organ failure, advanced age, hemorrhagic complications and irreversible central nervous system pathology. We presented a case of a 79-year-old Chinese lady without any pre-existing lung disease admitted to our hospital intensive care unit (ICU) after acute presentation of breathlessness and chest pain. After extensive workup, she was diagnosed with rapidly progressing acute interstitial pneumonia with ARDS and hypercapnia respiratory failure. The patient received lung protective strategies of mechanical ventilation and neuromuscular blockage therapy as per clinical guidelines. However, hypercapnia respiratory failure was refractory, and she was deemed not a good candidate for VV-ECMO support given her advanced age and high vasopressor requirements from shock. Alternative therapy with extracorporeal CO2 removal (ECCO2R) was considered and implemented. The patient received 12 days of ECCO2R paired with muscle paralysis, optimization of lung-protective mechanical ventilation and dialysis. Unfortunately, the patient still had refractory hypercapnic respiratory failure with dual vasopressor support despite prolonged therapy. Given failed and futile medical treatment, the family opted for withdrawal of care, a conservative approach, and comfort care, which led to her demise. The effectivity of extracorporeal CO2 removal may depend on disease burden, involvement and severity of the disease. There is insufficient data to make strong recommendations about its benefit-risk ratio for ECCO2R devices, and further studies and data would be required. Nonetheless, ECCO2R can be considered an alternative treatment for refractory hypercapnic respiratory failure patients who are unsuitable for initiating venovenous ECMO.

Keywords: extracorporeal CO2 removal (ECCO2R), acute respiratory distress syndrome (ARDS), acute interstitial pneumonia (AIP), hypercapnic respiratory failure

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Authors: Nutthawadee Jampanil, Kattareeya Kumthip, Niwat Maneekarn, Pattara Khamrin

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Rotavirus A is one of the leading causes of acute gastroenteritis in children younger than five years of age, especially in low-income countries in Africa and South Asia. Two live-attenuated oral rotavirus vaccines, Rotarix and RotaTeq, have been introduced into routine immunization programs in many countries and have proven highly effective in reducing the burden of rotavirus-associated morbidity and mortality. In Thailand, Rotarix and RotaTeq vaccines have been included in the national childhood immunization program since 2020. The objectives of this research are to conduct a molecular epidemiological study and to characterize rotavirus genotypes circulating in pediatric patients with acute diarrhea in Chiang Mai, Thailand, from 2020-2022 after the implementation of rotavirus vaccines. Out of 858 stool specimens, 26 (3.0%) were positive for rotavirus A. G3P[8] (23.0%) was detected as the most predominant genotype, followed by G1P[8] (19.2%), G8P[8] (19.2%), G9P[8] (15.3%), G2P[4] (7.7%), G1P[6] (3.9%), G9P[4] (3.9%), and G8P[X] (3.9%). In addition, the uncommon rotavirus strain G3P[23] (3.9%) was also detected in this study, and this G3P[23] strain displayed a genetic background similar to the porcine rotavirus. In conclusion, there was a dramatic change in the prevalence of rotavirus A infection and the diversity of rotavirus A genotypes in pediatric patients in Chiang Mai, Northern Thailand, in the rotavirus post-vaccination period. The finding obtained from this research contributes to a better understanding of rotavirus epidemiology after rotavirus vaccine introduction. Furthermore, the identification of unusual G and P genotype combination strains provides significant evidence for the potential interspecies transmission between human and animal rotaviruses.

Keywords: rotavirus, infectious disease, gastroenteritis, Thailand

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Authors: Wisit Wichitkosoom

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Objective:The majority of the hisk-risk patients (ST-elevation myocardial infarction (STEMI), Acute cerebrovascular accident, Sepsis, Acute Traumatic patient ) are admitted to district or lacal hospitals (average 1-1.30 hr. from Udonthani general hospital, Northeastern province, Thailand) without proper facilities. The referral system was support to early care and early management at pre-hospital stage and prepare for the patient data to higher hospital. This study assessed the reduction in treatment delay achieved by pre-hospital diagnosis and referral directly to Udonthani General Hospital. Methods and results: Four district or local hospitals without proper facilities for treatment the very high-risk patient were serving the study region. Pre-hospital diagnoses were established with the simple technology such as LINE, SMS, telephone and Fax for concept of LEAN process and then the telemedicine, by ambulance monitoring (ECG, SpO2, BT, BP) in both real time and snapshot mode was administrated during the period of transfer for safety transfer concept (inter-hospital stage). The standard treatment for patients with STEMI, Intracranial injury and acute cerebrovascular accident were done. From 1 October 2012 to 30 September 2013, the 892 high-risk patients transported by ambulance and transferred to Udonthani general hospital were registered. Patients with STEMI diagnosed pre-hospitally and referred directly to the Udonthani general hospital with telemedicine closed monitor (n=248). The mortality rate decreased from 11.69% in 2011 to 6.92 in 2012. The 34 patients were arrested on the way and successful to CPR during transfer with the telemedicine consultation were 79.41%. Conclusion: The proper innovation could apply for health care system. The very high-risk patients must had the closed monitoring with two-way communication for the “safety transfer period”. It could modified to another high-risk group too.

Keywords: safety transfer, telemedicine, critical patients, medical and health sciences

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Authors: Valdrina Ajeti, Icko Gjorgoski

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Stress can be a reason for some physiological and biological disorders in the body. The antioxidative defense system is necessary for the maintenance of redox homeostasis in organisms. Because of its antioxidant effect, alkaline water (AW) is the focus of scientific interest. Adding AW and co-treatment with sodium ascorbate (SA) is expected for the organism to act preventively to hyperthermic stress. To investigate the effect of AW and SA on glucose and cortisol levels during acute hyperthermic stress, white female Wistar laboratory rats, divided into three groups of 10 individuals, were exposed to heat for 80 min, for 21 days. Acute hyperthermic exposure at 41˚C was a cause for oxidative stress. The first group is the control group, the second group is treated with AW, and the third group with AW and SA. Plasma glucose levels were determined by colorimetric method and cortisol was measured using the enzyme-linked immunosorbent assay method. The comparison of the means was made using the Tukey test. Differences were considered significant at a level of p < 0.05. Our results show that levels of glucose and cortisol have been increased in the group treated with AW on the 21st day after treatment (p < 0.0001), but not on the 7th and 14th day as compared to the control group. Also, co-treatment of animals with AW and SA significantly increased the levels of glucose and cortisol on the 21st day after treatment showing a synergistic effect. The individual action of AW, as well as synergism with SA, caused a high protective effect on oxidative damage.

Keywords: alkaline water, sodium ascorbate, hyperthermic stress, glucose, cortisol

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Authors: L. L. Tundisi, A. Pessoa Jr., E. B. Tambourgi, E. Silveira, P. G. Mazzola

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L-asparaginase (L-ASNase) is the gold standard treatment for acute lymphoblastic leukemia that mainly affects pediatric patients; treatment increases survival from 20% to 90%. The characterization of other L-Asparaginases, apart from the most used from Escherichia coli and Erwinia chrysanthemi, has been reported, but the choice of the most appropriate is still under debate. This choice should be based on its pharmacokinetics, immune hypersensitivity, doses, prices, pharmacodynamics. The main factors influencing the antileukemic activity of ASNase are enzymatic activity, Km, glutaminase activity, clearance of the enzyme and development of resistance. However, most of the commercialized enzyme present an intrinsic glutaminase activity, which is responsible for some side effects. In this study, glutaminase free asparaginase produced from Aspergillus oryzae was precipitated in different percentages of ethanol (0–80%), until optimum ethanol concentration of 60% (w/w) was found. Following, precipitation of crude L-ASNase was performed in a single step, using 60% (w/w) ethanol, under constant agitation and temperature. It presented activity of 135.45 U/mg and after gel filtration chromatography with Sephadex G-the enzymatic activity was 322.02 U/mg. The apparent molecular mass of the purified L-ASNase fraction was estimated by 10% SDS-PAGE. Proteins were stained with Coomassie Brilliant Blue R-250. The molar mass range was from 10 kDa to 250 kDa. L-ASNase from Aspergillus oryzae was characterized aiming possible therapeutic use. Four different buffers (phosphate-citrate buffer pH 2.6 to 5.8; phosphate buffer pH 5.8 to 7.4; Tris - HCl pH 7.4 to 9.0; and carbonate buffer pH 9.8 to 10.6) were used to measure the optimum pH for L-ASNase activity. The optimum temperature for enzyme activity was measured at optimal pH conditions (Tris-HCl and phosphate buffer, pH 7.4) at different temperatures ranging from 5 to 55°C. All activities were calculated by quantifying the free ammonia, using the Nessler reagent. The kinetic parameters calculation, e.g. Michaelis-Menten constant (Km), maximum velocity (Vmax) and Hills coefficient (n), were performed by incubating the enzyme in different concentrations of the substrate at optimum conditions of pH and fitted on Hill’s equation. This glutaminase free asparaginase showed a low Km (3.39 mM and 3.81 mM) and enzymatic activity of 135.45 U/mg after precipitation with ethanol. After gel filtration chromatography it rose to 322.02 U/mg. Optimum activity was found between pH 5.8 - 9.0, best activity results with phosphate buffer pH 7.4 and Tris-HCl pH 7.4 and showed activity from 5°C to 55°C. These results indicate that L-ASNase from A. oryzae has the potential for human use.

Keywords: biopharmaceuticals, bioprocessing, bioproducts, biotechnology, enzyme activity, ethanol precipitation

Procedia PDF Downloads 291

Authors: Balakumar Pitchai, Xiang Llan Ang, Sunil Prajapati, Varatharajan Rajavel, Sundram Karupiah, Mohd Baidi Bahari

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The study examined the pretreatment and post-treatment effects of low-doses of fenofibrate and rosuvastatin in gentamicin-induced acute nephrotoxicity in rats. Gentamicin (100 mg/kg/day, i.p.) was administered to rats for 8 days. In the pretreatment protocol, low-dose fenofibrate (30 mg/kg/day, p.o.) or low-dose rosuvastatin (2 mg/kg/day, p.o.) treatments were started a day before the administration of gentamicin and continued for 8 days. In the post-treatment protocol, rats administered gentamicin were treated with low-dose fenofibrate (30 mg/kg/day, p.o.) or low-dose rosuvastatin (2 mg/kg/day, p.o.) for 6 days after the completion of 8 days protocol of gentamicin administration. Gentamicin-associated acute nephrotoxicity in rats was assessed in terms of biochemical analysis and renal histopathological studies. Gentamicin-administered rats showed marked renal functional changes as assessed in terms of a significant increase in serum creatinine and urea levels as compared to normal rats. The renal dysfunction noted in gentamicin administered rats was accompanied with elevated serum uric acid level as compared to normal rats while there was no significant change in lipid profile. Low-dose fenofibrate pretreatment in gentamicin-administered rats afforded a significant renal functional improvements and renoprotection while its post-treatment showed no significant renoprotection. On the other hand, pretreatment with low-dose rosuvastatin partially reduced gentamicin-induced increase in serum creatinine level, but its post-treatment did not afford renal functional improvements in gentamicin-administered rats. However, all pre and post-treatments with low-doses of fenofibrate or rosuvastatin significantly reduced the elevated serum uric acid concentration in gentamicin-administered rats. Renal histopathological analysis showed a discernible incidence of acute tubular necrosis in gentamicin-administered rats which were markedly reduced by low-dose fenofibrate or low-dose rosuvastatin pretreatments; but, not by their post-treatments. In conclusion, low-dose fenofibrate pretreatment considerably prevented gentamicin-induced acute tubular necrosis and renal functional abnormalities in rats while its post-treatment resulted in no significant renoprotective action. In spite of effective prevention of gentamicin-induced acute tubular necrosis, the pretreatment with low-dose rosuvastatin had only a partial and fractional protection on renal functional abnormalities. The post-treatment with low-dose rosuvastatin was ineffective in affording a renoprotection in gentamicin-administered rats.

Keywords: gentamicin-nephrotoxicity, low-dose fenofibrate, low-dose rosuvastatin, renoprotection

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Authors: Le Nguyen Binh, Luong Xuan Binh, Le Van Tuan, Tran Binh Duong, Truong Nguyen Khanh Hung, Do Le Hoang Son, Pham Quang Vinh, Hoang Quoc Huy, Nguyen Bach, Nguyen Quoc Khanh Le, Jiunn Horng Kang

Abstract:

Introduction: Traumatic hip dislocation is an emergency in young adult which can cause avascular necrosis of femoral head or osteoarthritis of hip joint. The reasons for these may be the loose body of bony or chondral fragments, which are difficult to be detected on CT scan or MRI. In those cases, Hip arthroscopy may be the method of choice for diagnosis and treatment of loose bodies in hip joint after traumatic dislocation. Methods: A case report is performed. A 55-year-old male patient was under hip arthroscopy to retrieve the loose body in the right hip joint. Results: The patient’s hip was reduced under anesthesia in the opeation room. Xray and CT scan post-reduction showed that his right hip was wide and a small fragment of femoral head (< 5mm) locking inside the joint. A hip arthroscopy was done to take the fragment out. Post-operation, the patient went under rehabilition. After 6 months, he can walk with full-weight bearing; no further dislocaion was noted, and the Harris score was 84 points. Conclusions: Although acute traumatic injury of hip joint is usually treated with open surgeries, these methods have many drawbacks, such as soft tissue destruction, blood-loss,….Despite its technical requirement, hip arthroscopy is less invasive and effective treatment. Therefore, it may be an alternative treatment for a traumatic hip injury and can be applied frequently in the near future.

Keywords: hip dislocation, hip arthroscopy, hip osteoarthritis, acute hip trauma

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Authors: Adenike Adesanya, Nonthaphat Wong, Xiang-Yun Lan, Shea Ping Yip, Chien-Ling Huang

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Background: The most challenging mutation of the oncokinase BCR-ABL protein T315I, which is commonly known as the “gatekeeper” mutation and is notorious for its strong resistance to almost all tyrosine kinase inhibitors (TKIs), especially imatinib. Therefore, this study aims to identify T315I-dependent downstream microRNA (miRNA) pathways associated with drug resistance in chronic myeloid leukemia (CML) for prognostic and therapeutic purposes. Methods: T315I-carrying K562 cell clones (K562-T315I) were generated by the CRISPR-Cas9 system. Imatinib-treated K562-T315I cells were subjected to small RNA library preparation and next-generation sequencing. Putative lncRNA-miRNA-mRNA networks were analyzed with (i) DESeq2 to extract differentially expressed miRNAs, using Padj value of 0.05 as cut-off, (ii) STarMir to obtain potential miRNA response element (MRE) binding sites of selected miRNAs on lncRNA H19, (iii) miRDB, miRTarbase, and TargetScan to predict mRNA targets of selected miRNAs, (iv) IntaRNA to obtain putative interactions between H19 and the predicted mRNAs, (v) Cytoscape to visualize putative networks, and (vi) several pathway analysis platforms – Enrichr, PANTHER and ShinyGO for pathway enrichment analysis. Moreover, mitochondria isolation and transcript quantification were adopted to determine the new mechanism involved in T315I-mediated resistance of CML treatment. Results: Verification of the CRISPR-mediated mutagenesis with digital droplet PCR detected the mutation abundance of ≥80%. Further validation showed the viability of ≥90% by cell viability assay, and intense phosphorylated CRKL protein band being detected with no observable change for BCR-ABL and c-ABL protein expressions by Western blot. As reported by several investigations into hematological malignancies, we determined a 7-fold increase of H19 expression in K562-T315I cells. After imatinib treatment, a 9-fold increment was observed. DESeq2 revealed 171 miRNAs were differentially expressed K562-T315I, 112 out of these miRNAs were identified to have MRE binding regions on H19, and 26 out of the 112 miRNAs were significantly downregulated. Adopting the seed-sequence analysis of these identified miRNAs, we obtained 167 mRNAs. 6 hub miRNAs (hsa-let-7b-5p, hsa-let-7e-5p, hsa-miR-125a-5p, hsa-miR-129-5p, and hsa-miR-372-3p) and 25 predicted genes were identified after constructing hub miRNA-target gene network. These targets demonstrated putative interactions with H19 lncRNA and were mostly enriched in pathways related to cell proliferation, senescence, gene silencing, and pluripotency of stem cells. Further experimental findings have also shown the up-regulation of mitochondrial transcript and lncRNA MALAT1 contributing to the lncRNA-miRNA-mRNA networks induced by BCR-ABL T315I mutation. Conclusions: Our results have indicated that lncRNA-miRNA regulators play a crucial role not only in leukemogenesis but also in drug resistance, considering the significant dysregulation and interactions in the K562-T315I cell model generated by CRISPR-Cas9. In silico analysis has further shown that lncRNAs H19 and MALAT1 bear several complementary miRNA sites. This implies that they could serve as a sponge, hence sequestering the activity of the target miRNAs.

Keywords: chronic myeloid leukemia, imatinib resistance, lncRNA-miRNA-mRNA, T315I mutation

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Authors: Noura El-Ahmady El-Naggar

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L-asparaginase is an important enzyme as therapeutic agents used in combination therapy with other drugs in the treatment of acute lymphoblastic leukemia in children. L-asparaginase producing actinomycete strain, NEAE-K, was isolated from soil sample and identified on the basis of morphological, cultural, physiological and biochemical properties, together with 16S rDNA sequence as new subsp. Streptomyces rochei subsp. chromatogenes NEAE-K and sequencing product (1532 bp) was deposited in the GenBank database under accession number KJ200343. The study was conducted to screen parameters affecting the production of L-asparaginase by Streptomyces rochei subsp. chromatogenes NEAE-K on solid state fermentation using Plackett–Burman experimental design. Sixteen different independent variables including incubation time, moisture content, inoculum size, temperature, pH, soybean meal+ wheat bran, dextrose, fructose, L-asparagine, yeast extract, KNO3, K2HPO4, MgSO4.7H2O, NaCl, FeSO4. 7H2O, CaCl2, and three dummy variables were screened in Plackett–Burman experimental design of 20 trials. The most significant independent variables affecting enzyme production (dextrose, L-asparagine and K2HPO4) were further optimized by the central composite design. As a result, a medium of the following formula is the optimum for producing an extracellular L-asparaginase by Streptomyces rochei subsp. chromatogenes NEAE-K from solid state fermentation: g/L (soybean meal+ wheat bran 15, dextrose 3, fructose 4, L-asparagine 8, yeast extract 2, KNO3 1, K2HPO4 2, MgSO4.7H2O 0.5, NaCl 0.1, FeSO4. 7H2O 0.02, CaCl2 0.01), incubation time 7 days, moisture content 50%, inoculum size 3 mL, temperature 30°C, pH 8.5.

Keywords: streptomyces rochei subsp. chromatogenes neae-k, 16s rrna, identification, solid state fermentation, l-asparaginase production, plackett-burman design, central composite design

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