Search results for: gene therapy

Authors: Małgorzata Wrzosek, Nina Baruch, Beata Jabłonowska-Lietz

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Background: The fat mass & obesity-associated (FTO) gene is linked to an increased risk of obesity. However, the relation between rs9939609 and eating behaviors or energy expenditure is not fully elucidated. The aim of this study was to investigate the relationship between the rs9939609 polymorphism of the FTO gene and the postprandial satiety, sweets intake, physical activity and depressive symptoms in patients with obesity. Methods: The study group consisted of 585 subjects with a BMI of 42.97.0 kg/m². The rs9939609 polymorphism of the FTO gene was examined using real time – PCR method. The severity of depressive symptoms was assessed with the Beck Depression Inventory (BDI-II). Information was obtained about demographics, eating habits and lifestyle. Results: More than half (63.5%) of the patients reported consumption of sweets between main meals and 30% declared high and very high postprandial satiety and the frequency of TA/AA carriers in rs9939609 (FTO) compared with TT carriers was similar. Significantly lower BDI-II scores were found in subjects with higher level of physical activity and it was seen amongst patients with the AA and AT genotypes of the FTO rs9939609 polymorphism. Conclusion: Obesity is a highly heritable trait, but eating habits also appear as major factors affecting obesity development.

Keywords: FTO polymorphism, physical activity, obesity, depression, postprandial satiety, sugary foods, sweets

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Authors: C. Pearcy, C. Rees

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Cognitive behavioural therapy (CBT) and exposure and response prevention (ERP) are currently the most efficacious treatments for Obsessive-compulsive disorder (OCD). Many clients, however, remain symptomatic following treatment. As a result, refusal of treatment, withdrawal from treatment, and partial adherence to treatment are common amongst ERP. Such limitations have caused few professionals to actually engage in ERP therapy, which has warranted the exploration of alternative treatments. This study evaluated an online self-help treatment program for OCD (the OCD Doctor Online); a 4-week Metacognitive Therapy (MCT) program which has implemented strategies from Wells’ Metacognitive model of OCD. The aim of the present study was to investigate whether an online self-help treatment using MCT would reduce symptoms of OCD, reduce unhelpful metacognitions and improve quality of life. Treatment effectiveness was assessed using a case series methodology in 3 consecutively referred individuals. At post-treatment, all participants showed reductions in unhelpful metacognitive beliefs (MCQ-30) and improvements in quality of life (Q-LES-Q), which were maintained through to 4 week follow-up. Two of the three participants showed reductions in OCD symptomology (OCI-R), which were further reduced at 4-week follow-up. The present study suggests that internet-based self-help treatment may be an effective means of delivering MCT to adults with OCD.

Keywords: internet-based, metacognitive therapy, obsessive-compulsive disorder, self-help

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Authors: R. D. Esposito, V. M. Barberá, P. García Morales, P. Dorado Rodríguez, J. Sanz, M. Fuentes, D. Planes Meseguer, M. Saceda, L. Fernández Fornos, M. P. Ventero

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Results obtained by our group on glioblastoma multiforme (GBM) primary cultures , show a dramatic potentiation of radiation effects when 2 units/ml of D-amino acid oxidase (DAO) enzyme are added, free or immobilized in magnetic nanoparticles, to irradiated samples just after the irradiation. Cell cultures were exposed to radiation doses of 7Gy and 15Gy of 6 MV photons from a clinical linear accelerator. At both doses, we observed a clear enhancing effect of radiation-induced damages due to the addition of DAO.

Keywords: D-amino Acid Oxidase (DAO) enzyme, magnetic particles, nanotechnology, radiation therapy enhancement

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Authors: Ahmad Ali Shahid, M Shakil Shaukat

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Agriculture is the backbone of economy of Pakistan and Cotton is the major agricultural export and supreme source of raw fiber for our textile industry. To combat against the developing resistance in the target insects and combating these challenges wholesomely, a novel combination of pyramided/stacked genes was conceptualized and later realized, through the means of biotechnology i.e., transformation of three genes namely, Cry1Ac, Cry2A, and EPSP synthase (glyphosate tolerant) genes in the locally cultivated cotton variety. The progenies of the transformed plants were successfully raised and screened under the tunnel conditions for two generations and the present study focused on the screening of plants which were confirmed for containing all of these three genes and their expressions. Initially, the screening was done through glyphosate spray assay and the plants which were healthy and showed no damage on leaves were selected after 07 days of spray. In the laboratory, the DNA of these plants were isolated and subjected to amplification of the three genes. Thus, seventeen out of twenty were confirmed positive for Cry1Ac gene and ten out of twenty were positive for Cry2A gene and all twenty were positive for presence of EPSP synthase gene. Then, the ten plant samples which were confirmed with presence of all three genes were subjected to expression analysis of these proteins through ELISA. The results showed that eight out of ten plants were actively expressing the three transgenes. Real-time PCR was also done to quantify the expression levels of the EPSP synthase gene. Finally, eight plants were confirmed for the presence and active expression of all three genes in T3 generation of the triple gene transformed cotton. These plants may be subjected to T4 generation to develop a new stable variety in due course of time.

Keywords: agriculture, cotton, transformation, cry genes, ELISA, PCR

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Authors: Rasime Kalkan, Emine Ikbal Atli, Ali Arslantaş, Muhsin Özdemir, Sevilhan Artan

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Glioblastoma (WHO grade IV), is the malignant form of brain tumor, the genetic background of the GBM is highly variable. The tumor mass of a GBM is multilayered and every tumor layer shows distinct characteristics with a different cell population. The treatment planning of GBM should be focused on the tumor genetic characteristics. We screened primary glioblastoma multiforme (GBM) in a population-based study for MGMT and RARβ methylation and IDH1 mutation correlated them with clinical data and treatment. There was no correlation between MGMT-promoter methylation and overall survival. The overall survival time of the patients with methylated RARβ was statically (OS;p<0,05) significance between the patients who were treated with chemotherapy and radiotherapy. Here we showed the status of IDH1 gene associatied with younger age. We demonstrated that the together with MGMT gene the RARβ gene should be used as a potantial treatment decision marker for GBMs.

Keywords: RARβ, primary glioblastoma multiforme, methylation, MGMT

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Authors: Zahra Hassani

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Background and Purpose: Poststroke depression (PSD) is one of the complications of a stroke that reduces the patient's chance of recovery, becomes irritable, and changes personality. Cognitive rehabilitation is one of the non-pharmacological methods that improve deficits such as attention, memory, and symptoms of depression. Therefore, the purpose of the present study is to evaluate the Efficacy of Cognitive Rehabilitation Therapy on Poststroke Depression among Survivors of stroke. Method: In this study, a systematic review of the databases Google Scholar, PubMed, Science Direct, Elsevier between the years 2015 and 2019 with the keywords cognitive rehabilitation therapy, post-stroke, depression Search is done. In this process, studies that examined the Efficacy of Cognitive Rehabilitation Therapy on Poststroke Depression among Survivors of stroke were included in the study. Results: Inclusion criteria were full-text availability, interventional study, and non-review articles. There was a significant difference between the articles in terms of the indices studied, sample number, method of implementation, and so on. A review of studies have shown that cognitive rehabilitation therapy has a significant role in reducing the symptoms of post-stroke depression. The use of these interventions is also effective in improving problem-solving skills, improving memory, and improving attention and concentration. Conclusion: This study emphasizes on the development of efficient and flexible adaptive skills through cognitive processes and its effect on reducing depression in patients after stroke.

Keywords: cognitive therapy, depression, stroke, rehabilitation

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Authors: Ghaleb Bin Huraib, Fahad Al Harthi, Mohammad Mustafa, Abdulrahman Al-Asmari

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Introduction: Vitiligo is an acquired pigmentary skin disorder with the regional disappearance of melanocytes. Vitiligo affects 0.1 to 2% of the global population, and the incidence varies substantially depending on ethnicity. Glutathione S-transferase (GST) is a multigene family of enzymes that detoxify oxidative stress products. The oxidative stress-related GSTM1/GSTT1 genes deletion may cause epidermal melanocytes destruction and the development of vitiligo. Hence, the present study aimed to investigate the association of GST gene polymorphisms with vitiligo in the Saudi population, if any. Materials and Methods: The present study includes 129 vitiligo cases and 130 age-matched healthy controls. The proportion of male and female patients with vitiligo is almost equal. The multiplex polymerase chain reaction (PCR) method was used for polymorphic analysis. Results: Increased odds of generalized vitiligo was observed with the null genotypes of GSTT1- gene (OR = 1.91, 95% CI = 1.07-3.42, p = 0.019). The possible genetic combinations of GSTM1/GSTT1 and their genotypic distribution showed the frequency of GSTM1+/GSTT1+ 62/130 (47.69%) and GSTM1-/GSTT1+ 52/130 (40.00%) were higher in controls than in cases 44/129 (34.11%), 43/129 (33.34%), respectively while GSTM1+/GSTT1- and GSTM1-/GSTT1- null genotypes were higher 22/129 (17.05%) and 20/129 (15.50%) in vitiligo patients as compared to controls 11/130 (8.46%), 5/130 (3.84%), respectively. The strength of association of different genetic combinations with cases have shown GSTM1+/GSTT1- (OR = 2.81, 95% CI = 1.24-6.40, p = 0.009) and GSTM1-/GSTT1- (OR = 5.63, 95% CI = 1.96 - 16.16, p = 0.0004) were significantly higher in vitiligo cases as compared to controls. We did not observe any significant association of age and gender of patients with GST gene polymorphisms. Conclusions: The GSTT1-, GSTM1+/GSTT1- and GSTM1-/GSTT1- null genotypes were significantly associated with vitiligo. These genetic polymorphisms may be the associative genetic risk factor for vitiligo among Saudis. It could be used as a genetic marker for screening vitiligo patients among Saudis. Further studies on GSTs gene polymorphism in larger sample sizes from different geographical areas and ethnicity are needed to strengthen the present findings.

Keywords: vitiligo, GSTM1, GSTT1, gene polymorphism, oxidative stress

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Authors: Yasir Karabacak

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Objective: The aim of periodontal therapy is to control and eliminate inflammation in order halt disease progression. The initial periodontal therapy (IPT) including scaling and root planing (SRP) can control periodontal disease in most cases of periodontitis; also maintaining good oral hygiene by the patient is fundamental. The aim of this case report is to present IPT and to present 3-month follow-up results in a patient with periodontitis. Materials and Methods IPT of a 63-year-old non-smoker male with generalized periodontitis is presented. The patient had no history of systemic disease. The intraoral examination reveals marked gingival inflammation as well as plaque accumulation and significant calculus deposits. On radiographic examination, severe bone loss was evident. The patient was diagnosed with generalized advanced periodontitis. Initial periodontal therapy including oral hygiene instructions and quadrant-based SRP under local anesthesia was performed using hand and ultrasonic instruments. No antibiotics were prescribed. The patient was recalled 4 weeks after IPT. Results Favorable clinical improvement was obtained. Gingival inflammation was resolved significantly. A reduction of the mean probing depth from 2.4 mm at baseline to 1.9 mm was observed. The patient presented with a good standard of oral hygiene. The plaque scores decreased from 54.0% at baseline to 17.0%. In addition, the percentage of sites with bleeding on probing decreased from 80.0% at baseline to 44.0%. The patient was scheduled for maintenance therapy every three months. Conclusion: The level of oral hygiene has a great impact on periodontal treatment outcome and supports periodontal therapy properly.

Keywords: initial periodontal, therapy and follow-up in a periodontitis, patient, a case report

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Authors: Gileade P. Freitas, Helena B. Lopes, Alann T. P. Souza, Paula G. F. P. Oliveira, Adriana L. G. Almeida, Paulo G. Coelho, Marcio M. Beloti, Adalberto L. Rosa

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Bone tissue presents great capacity to regenerate when injured by trauma, infectious processes, or neoplasia. However, the extent of injury may exceed the inherent tissue regeneration capability demanding some kind of additional intervention. In this scenario, cell therapy has emerged as a promising alternative to treat challenging bone defects. This study aimed at evaluating the effect of local injection of bone marrow-derived mesenchymal stem cells (BM-MSCs) and adipose tissue-derived mesenchymal stem cells (AT-MSCs) on bone regeneration of rat calvaria defects. BM-MSCs and AT-MSCs were isolated and characterized by expression of surface markers; cell viability was evaluated after injection through a 21G needle. Defects of 5 mm in diameter were created in calvaria and after two weeks a single injection of BM-MSCs, AT-MSCs or vehicle-PBS without cells (Control) was carried out. Cells were tracked by bioluminescence and at 4 weeks post-injection bone formation was evaluated by micro-computed tomography (μCT) and histology, nanoindentation, and through gene expression of bone remodeling markers. The data were evaluated by one-way analysis of variance (p≤0.05). BM-MSCs and AT-MSCs presented characteristics of mesenchymal stem cells, kept viability after passing through a 21G needle and remained in the defects until day 14. In general, injection of both BM-MSCs and AT-MSCs resulted in higher bone formation compared to Control. Additionally, this bone tissue displayed elastic modulus and hardness similar to the pristine calvaria bone. The expression of all evaluated genes involved in bone formation was upregulated in bone tissue formed by BM-MSCs compared to AT-MSCs while genes involved in bone resorption were upregulated in AT-MSCs-formed bone. We show that cell therapy based on the local injection of BM-MSCs or AT-MSCs is effective in delivering viable cells that displayed local engraftment and induced a significant improvement in bone healing. Despite differences in the molecular cues observed between BM-MSCs and AT-MSCs, both cells were capable of forming bone tissue at comparable amounts and properties. These findings may drive cell therapy approaches toward the complete bone regeneration of challenging sites.

Keywords: cell therapy, mesenchymal stem cells, bone repair, cell culture

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Authors: Bilge Meral Koc, Elvan Yilmaz Akyuz, Tugce Ozlu

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This study aimed to discover the effects of coconut oil intake and diet therapy on anthropometric measurements, biochemical findings and irisin levels in overweight individuals. Materials and Methods: Overweight individuals (n=44, 19-30 years) without any chronic disease were included. In this randomized controlled crossover study, the participants were divided into two groups (Group 1: 23 people, Group 2: 21 people). In the first phase, Group 1 received diet therapy to lose 0.5-1 kg of weight per week and 20 mL of coconut oil/day, while Group 2 only received diet therapy. In the second phase, Group 1 received diet therapy while Group 2 received diet therapy and 20 mL of coconut oil/day. Anthropometric measurements were taken four times. Irisin was measured four times by enzyme-linked immunosorbent (ELISA) method and other biochemical findings were measured twice. Statistical analysis was made on SPSS 20. Results: The irisin level decreased significantly when the participants only took coconut oil (p≤0.05). There was a significant decrease in the participants' body weight, body mass index (BMI) level and body fat percentage (p≤0.01). Insulin, total cholesterol, low density lipoproteins (LDL) cholesterol, and triglyceride (TG) levels of all participants decreased significantly (p≤0.05). There was no significant difference in irisin level due to body weight loss (p≤0.05); coconut oil provided a significant decrease in irisin level (p≤0.05). Conclusion: Diet therapy and weight loss did not have an effect on irisin level, but coconut oil alone was found to reduce irisin level. Coconut oil had no impact on anthropometric and biochemical findings.

Keywords: coconut oil, diet therapy, irisin, overweight

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Authors: K. M. Saif Ur Rahman, Razib Mamun, Himica Arjuman, Fida Al Shams

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Introduction: Autism spectrum disorder (ASD) has become an emerging neurodevelopmental disorder with increasing prevalence. It has become an important public health issue globally. Many approaches including speech and language therapy (SLT), occupational therapy, behavioral therapy etc. are being applied for the betterment of the ASD patients. This study aims to describe the characteristics of ASD patients and perception of caregiver regarding SLT in Bangladesh. Methods: This cross-sectional study was conducted in a therapy and rehabilitation center at Dhaka city. Caregivers of 48 ASD patients responded regarding their perception of SLT and characteristics of patients. Results: Among 48 ASD patients, 56.3% were between 3 to 5 years age group with a male predominance (87.5%). More than half of the participants (56.3%) initiated SLT at the age of 1-3 years and the majority (43.8%) were taking SLT for less than 1 year. Majority of the patients (64.6%) were taken to a physician for healthcare as a first contact of which 29.2% were referred to SLT by physicians. More than half (56.3%) of the caregivers were moderately satisfied with SLT and most of them (62.5%) mentioned moderate improvement through SLT. Improvement rate was 10-15% in specific symptoms such as eye contact, complex mannerism, pointing, imitation etc. Conclusion: This study reveals the self-reported perception of caregivers on SLT. Despite reported improvements, more exploration of different approaches and intervention for management of ASD is recommended.

Keywords: ASD, characteristics, SLT, Bangladesh

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Authors: Christopher G. Harris, Guy D. Eslick

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Purpose: Lobular carcinoma in situ (LCIS) is a known risk factor for breast cancer of unclear significance when detected in association with invasive carcinoma. This meta-analysis aims to determine the impact of LCIS on local recurrence risk for individuals with breast cancer treated with breast conservation therapy to help guide appropriate treatment strategies. Methods: We identified relevant studies from five electronic databases. Studies were deemed suitable for inclusion where they compared patients with invasive breast cancer and concurrent LCIS to those with breast cancer alone, all patients underwent breast conservation therapy (lumpectomy with adjuvant radiation therapy), and local recurrence was evaluated. Recurrence data were pooled by use of a random effects model. Results: From 1488 citations screened by our search, 8 studies were deemed suitable for inclusion. These studies comprised of 908 cases and 10638 controls. Median follow-up time was 90 months. There was a significantly increased overall risk of local breast cancer recurrence for individuals with LCIS in association with breast cancer following breast conservation therapy [pOR 1.87; 95% CI 1.14-3.04; p = 0.012]. The risk of local recurrence was non-significantly increased at 5 [pOR 1.09; 95% CI 0.48-2.48; p = 0.828] and 10 years [pOR 1.90; 95% CI 0.89-4.06; p = 0.096]. Conclusions: Individuals with LCIS in association with invasive breast cancer have an increased risk of local recurrence following breast conservation therapy. This supports consideration of aggressive local control of LCIS by way of completion mastectomy or re-excision for certain high-risk patients.

Keywords: breast cancer, breast conservation therapy, lobular carcinoma in situ, lobular neoplasia, local recurrence, meta-analysis

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Authors: Sidika McNeil

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Borderline personality disorder (BPD) is a personality disorder that has been found to have strong origins in childhood trauma. One of the key symptoms of BPD is an association with irregular moods swings, as well as suicidal ideation (SI). Owing to the typically severe trauma patients experience during childhood, it is hard for them to control their emotions and thus makes it hard to emotionally regulate. It is then very common for those suffering from BPD to turn to unhealthy coping mechanisms, such as substance use, unhealthy relationships, and more, often unsuccessfully creating experiences that facilitate safety which leads to further negative experiences. With the high suicide rating among children, adolescents, and teens, and an ever-increasing number of children being diagnosed with BPD, it is very important that more research is done to find further treatments for patients who are currently suffering. Methods: Utilizing data found in prior studies, this paper will analyze the literature to focus on a comprehensive treatment plan for those with DBT. It is currently suggested that with the use of dialectical behavioral therapy (DBT), a therapy that focuses on changing negative thinking patterns and pushes for more positive ones is helpful for treatment for those with BPD. Though this therapy is not a cure to BPD, it does help mitigate the risk; this essay will explore other options that can further the treatment process, such as cognitive analytical therapy (CAT), which focuses on delving into the past to find the root causes of an issue to create coping strategies and harm reduction, a type of therapy used to aid patients in lowering the use of substances without complete cessation. Results: The research provides enough evidence to link between the treatment of BPD with the utilization of CAT.

Keywords: borderline personality disorder, cognitive analytical therapy, dialectical behavioral therapy, harm reduction, suicidal ideation

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Authors: Hassan Abo-Salem, Huang Xiaolin

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Background and Purpose: The effectiveness of mirror therapy (MT) has been investigated in acute hemiplegia. The present study examines whether MT, given during chronic stroke, was more effective in promoting motor recovery of the lower extremity and walking speed than standard rehabilitation alone. Methods: The study enrolled 30 patients with chronic stroke. Fifteen patients each were assigned to the treatment group and the control group. All patients received a conventional rehabilitation program for a 4-week period. In addition to this rehabilitation program, patients in the treatment group received mirror therapy for 4 weeks, 5 days a week. Main measures: Passive ankle joint dorsiflexion range of motion, gait speed, Brunnstrom stages of motor recovery, plantarflexor muscle tone by Modified Ashworth Scale. Results: Results: No significant difference was found in the outcome measures among groups before treatment. When compared with standard rehabilitation, mirror therapy improved Ankle ROM, Brunnstrom stages and waking speed (p < 0.05). However, there were no significant differences between two groups on MAS (P > 0.05). Conclusions: Mirror therapy combined with a conventional stroke rehabilitation program enhances lower-extremity motor recovery and walking speed in chronic stroke patients.

Keywords: mirror therapy, stroke, MAS, walking speed

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Authors: Naomi Seidu, Edward Poluyi, Chibuikem Ikwuegbuenyi, Eghosa Morgan

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The current poor prognosis of glioblastoma has called for the need for an improvement in treatment methods in order to improve its survival rate. Despite the different interventions currently available for this tumor, the average survival is still only a few months. (12-15). The aim is to create a more favorable prognosis and have a reduction in the resistance to treatment currently being experienced, even with surgical interventions and chemotherapy. From the available literature, there is a relationship between the presence of HOX genes (Homeobox genes) and glioblastoma, which could be attributable to the increasing treatment resistance. Hence silencing these genes can be a key to improving survival rates of glioblastoma. A series of studies have highlighted the role that HOX genes play in glioblastoma prognosis. Promotion of human glioblastoma initiation, aggressiveness, and resistance to Temozolomide has been associated with HOXA9. The role of HOX gene expression in cancer stem cells should be studied as it could provide a means of designing CSC-targeted therapies, as CSCs play a part in the initiation and progression of solid tumors.

Keywords: GBM- glioblastoma, HOXA gene- homeobox genes cluster, signaling pathways, temozolomide

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Authors: Tai-Young Park, Yangjin Park

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Introduction: This study analyzed a family therapy case involving a female client in her thirties with panic disorder (PD) in South Korea. We identified five stages of the psychological process in the development of PD and examined external situations, family dynamics, and psychological experiences at each stage. Method: The client, mother, sister, and husband participated in therapy. Researchers analyzed the transcripts, notes, and video recordings of the therapy sessions. A thematic analysis was used to examine the data and display our findings using a network. Results: The developmental process of PD was as follows: (1) formation of anxiety, (2) sheltered life, (3) crisis, (4) loss of safe haven, and (5) inner breakdown. Conclusion: The family dynamics that developed as a result of coping with external situations in each stage contributed to clients’ psychological experiences. These psychological experiences triggered anxiety, which led to the development of PD. Moreover, this study empirically suggests that family dynamics can be associated with a person’s internal experiences that could lead to PD. Our findings highlight the significance of functional family dynamics and coping patterns when facing difficult external situations or crises.

Keywords: developmental process, family therapy, panic disorder, psychological dynamics

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Authors: Ibrahim M. I. Hamoda, Ahmed R. Z. Baghdadi, Mohammed K. Mohamed, Nawal A. Abu-Shady

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Background: Most of patients with vestibular dysfunction suffering from balance disorders, Abnormality in balance increase effort and exertion which affect the independency, so this study might be a guide in managing balance problem and consequently improve walking with less exertion and maximum function. Purpose: to analyze and discuss the effect of transcranial magnetic therapy on balance in patients with vestibular dysfunction. Methods: forty subjects from both sexes were classified to divided randomly into two equal groups; Group I study group: this group received transcranial magnetic therapy, with a selected physical therapy program for improving balance and vestibular disorders (Balance training, Cawthorne-Cooksey Exercises) and group II (control group): this group received a selected physical therapy program as group I without transcranial magnetic therapy. This treatment procedure will be applied three times weekly for three months. The mean age was 54.53±3.44 and 55.33±2.32 years and BMI 35.7±3.03 and 35.73±1.03 kg/m2 for group I and II respectively. The Biodex Balance System, Berge balances scale (BBS) and brain MRI were used for assessment. Assessments were conducted before and after treatment. The treatment program for group I included balance training, Cawthorne-Cooksey Exercises and pulsed magnetic therapy (Parameters used in the program of 20 minutes, Intensity 2 gausses, Frequency 1 Hz). This selected program was done in approximately one hour every other day for three month. The treatment program group II Patients received the same program as group A without transcranial magnetic therapy. Results: The One-way ANOVA revealed that there were no significant differences in BBS scores, overall balance index, Anterior / posterior balance index, Medial / lateral balance index and dynamic limits of stability between both groups. Moreover, the BBS scores increased and overall balance index, Anterior / posterior balance index, Medial / lateral balance index and dynamic limits of stability decreased significantly after treatment in group I and II compared with before treatment. Interpretation/Conclusion: Adding pulsed magnetic therapy to balance training, Cawthorne-Cooksey Exercises has no effect on static and dynamic balance in patients with balance problems due to benign positional paroxysmal vertigo.

Keywords: balance, transcranial magnetic therapy, vestibular dysfunction, biomechanic

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Authors: Khaled Khleifat, Muayyad Abboud, Ahmad Almustafa

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The effect of metabolic inhibitor/uncoupler(s) (CCCP and NaN3) and sulfhydryl reagents (dithiothreitol, 2 mercaptoethanol glutathione) on cadmium uptake was investigated in Enterobacter aerogenes strains. They include a transformed strain bearing the Vitreoscillahemoglobin gene, vgb as well as control strains that lack this transformed gene. The vgb-harboring strains showed better uptake of cadmium than vgb-lacking strains. Under low aeration, there was 2 fold enhancement of Cd+2 uptake in vgb-harboring strains compared with 1.6-fold enhancement under high aeration. The CCCP caused 36, 40 and 58% inhibition in cadmium uptake of parental, pUC9 harboring and VHb expressing cells, respectively. Similarly, the sodium azide exerted 44, 38 and 55% inhibition in Cd+2 uptake of parental, pUC9 harboring and VHb expressing cells, respectively. Less extensive inhibition of Cd+2 uptake in the range of 11 to 39% was observed with sulfhydryl reagents.

Keywords: bacterial hemoglobin, VHb, Cd uptake, biosorption

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Authors: H. K. Ratre, M. Roy, S. Roy, M. S. Parmar, V. Bhagat

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Mastitis is a common problem of dairy industries. Reduction in milk production and an irreparable damage to the udder associated with the disease are common causes of culling of dairy cows. Milk from infected animals is not suitable for drinking and for making different milk products. So, it has a major economic importance in dairy cattle. The aims of this study were to investigate the bacteriological panorama in milk from udder quarters with subclinical mastitis and to carried out for the molecular characterization of the major isolated organisms, from subclinical mastitis-affected cows in and around Durg and Rajnandgaon district of Chhattisgarh. Isolation and identification of bacteria from the milk samples of subclinical mastitis-affected cows were done by standard and routine culture procedures. A total of 78 isolates were obtained from cows and among the various bacteria isolated, Staphylococcus spp. occupied prime position with occurrence rate of 51.282%. However, other bacteria isolated includeStreptococcus spp. (20.512%), Micrococcus spp. (14.102%), E. coli (8.974%), Klebsiela spp. (2.564%), Salmonella spp. (1.282%) and Proteus spp. (1.282%). Staphylococcus spp. was isolated as the major causative agent of subclinical mastitis in the studied area. Molecular characterization of Staphylococus aureusisolates was done for genetic expression of the virulence genes like ‘nuc’ encoding thermonucleaseexoenzyme, coa and spa by PCR amplification of the respective genes in 25 Staphylococcus isolates. In the present study, 15 isolates (77.27%) out of 20 coagulase positive isolates were found to be genotypically positive for ‘nuc’ where as 20 isolates (52.63%) out of 38 CNS expressed the presence of the same virulence gene. In the present study, three Staphylococcus isolates were found to be genotypically positive for coa gene. The Amplification of the coa gene yielded two different products of 627, 710 bp. The amplification of the gene segment encoding the IgG binding region of protein A (spa) revealed a size of 220 and 253bp in twostaphylococcus isolates. The X-region binding of the spa gene produced an amplicon of 315 bp in one Staphylococcal isolates. Staphylococcus aureus was found to be major isolate (51.28%) responsible for causing subclinical mastitis in cows which also showed expression of virulence genesnuc, coa and spa.

Keywords: mastitis, bacteria, characterization, expression, gene

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Authors: Dina Athariah, Desriani Desriani, Bugi Ratno Budiarto, Abinawanto Abinawanto, Dwi Wulandari

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Breast cancer is a regulated by many genes. Defect in PIK3CA gene especially at position of exon 9 (E542K and E545K), called hot spot mutation induce early transformation of breast cells. The early detection of breast cancer based on mutation profile of this hot spot region would be hampered by the existence of pseudogene, marked by its substitution mutation at base 1658 (E545A) and deletion at 1659 that have been previously proven in several cancers. To the best of the authors’ knowledge, until recently no studies have been reported about pseudogene phenomenon in breast cancer. Here, we reported PCR optimization to to obtain true exon 9 of PIK3CA gene from its pseudogene hence increasing the validity of data. Material and methods: two genomic DNA with Dev and En code were used in this experiment. Two pairs of primer were design for Standard PCR method. The size of PCR products for each primer is 200bp and 400bp. While other primer was designed for Nested-PCR followed with DNA sequencing method. For Nested-PCR, we optimized the annealing temperature in first and second run of PCR, and the PCR cycle for first run PCR (15x versus 25x). Result: standard PCR using both primer pairs designed is failed to detect the true PIK3CA gene, appearing a substitution mutation at 1658 and deletion at 1659 of PCR product in sequence chromatogram indicated pseudogene. Meanwhile, Nested-PCR with optimum condition (annealing temperature for the first round at 55oC, annealing temperatung for the second round at 60,7oC with 15x PCR cycles) and could detect the true PIK3CA gene. Dev sample were identified as WT while En sample contain one substitution mutation at position 545 of exon 9, indicating amino acid changing from E to K. For the conclusion, pseudogene also exists in breast cancer and the apllication of optimazed Nested-PCR in this study could detect the true exon 9 of PIK3CA gene.

Keywords: breast cancer, exon 9, hotspot mutation, PIK3CA, pseudogene

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Authors: Hend Almaseb

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​The motivational model proposed by Self-Determination Theory provided an explanation for clients’ motivation for therapy. Among a sample of 78 inpatient residents in the Addiction Treatment Center, this study examined the relationship between three types of motivation (Autonomous, Controlled, and Amotivation) and each of the following variables: Age, Marital Status, Educational Level of Participant, and Number of Years of Addiction. In addition, the study investigated whether or not the participants are motivated to receive therapy. The results showed 1) a significant relationship between Controlled Motivation and the following variables: Age, Marital Status, and Number of Years of Addiction; 2) a significant relationship between Autonomous Motivation and Number of Years of Addiction; and a significant relationship between Educational Level and Amotivation. The results also illustrated that the participants of this study were not motivated to seek therapy.

Keywords: addiction, clinical social work, motivation, self-determination

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Authors: Surajit Bhattacharya, Daniel Veltri, Atit A. Patel, Daniel N. Cox

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miRNAs have emerged as key post-transcriptional regulators of gene expression, however identification of biologically-relevant target genes for this epigenetic regulatory mechanism remains a significant challenge. To address this knowledge gap, we have developed a novel tool in R, Intra-miR-ExploreR, that facilitates integrated discovery of miRNA targets by incorporating target databases and novel target prediction algorithms, using statistical methods including Pearson and Distance Correlation on microarray data, to arrive at high confidence intragenic miRNA target predictions. We have explored the efficacy of this tool using Drosophila melanogaster as a model organism for bioinformatics analyses and functional validation. A number of putative targets were obtained which were also validated using qRT-PCR analysis. Additional features of the tool include downloadable text files containing GO analysis from DAVID and Pubmed links of literature related to gene sets. Moreover, we are constructing interaction maps of intragenic miRNAs, using both micro array and RNA-seq data, focusing on neural tissues to uncover regulatory codes via which these molecules regulate gene expression to direct cellular development.

Keywords: miRNA, miRNA:mRNA target prediction, statistical methods, miRNA:mRNA interaction network

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Authors: Omid Azimaraghi, Noushin Khazaei

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Background: Fluid therapy is an important aspect of the perioperative period and a major challenge for anesthesiologists. To authors best knowledge, there is a lack of strong guidance and evidence regarding the optimal approach to fluid therapy. Therefore a variety of medical devices have been introduced to help physicians. In this study, we aimed to evaluate the effectiveness of pleth variability index in guiding fluid therapy in different patient positions. Materials and Methods: Inclusion criteria consisted of patients aged 18-50 years old and classified as American Society of Anesthesiologists physical status I and II, who were candidates for elective thyroidectomy surgery. In total, 36 patients meeting the inclusion criteria were enrolled in the study. After induction of anesthesia and start of mechanical ventilation Pleth variability index was measured in the supine position, then patients were placed in Trendelenburg and reverse Trendelenburg position (30 degrees, 5 minutes); Pleth Variability Index has measured again in the mentioned positions. Results: Mean PVI (Pleth Variability Index) in the supine position was 14.3 ± 3.7 in comparison to 21.5 ± 4.3 in the reverse Trendelenburg position. The mean PVI in Trendelenburg position was 9.1 ± 2.0 in Trendelenburg position (p < 0.05). Conclusion: In conclusion, we found that Pleth Variability Index varies with patient position and this should be taken into account when using this index during fluid therapy.

Keywords: fluid therapy, Pleth Variability Index, position, surgery

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Authors: Mehrdad Sheikhvatan, Mohammad Ali Boroumand, Mehrdad Behmanesh, Shayan Ziaee

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Contradictory results have been obtained regarding the role of integrin, beta 3 (ITGB3) gene polymorphisms in occurrence of acute myocardial infarction (MI) in patients with coronary artery disease (CAD). Hence, we aimed to assess the association between 1565C/T polymorphism of ITGB3 gene and increased risk for acute MI in patients who suffered premature CAD in Iranian population. Our prospective study included 1000 patients (492 men and 508 women aged 21 to 55 years) referred to Tehran Heart center during a period of four years from 2008 to 2011 with the final diagnosis of premature CAD and classified into two groups with history of MI (n = 461) and without of MI (n = 539). The polymorphism variants were determined by PCR-RFLP technique by entering 10% of randomized samples and then genotyping of the polymorphism was also conducted by High Resolution Melting (HRM) method. Among study samples, 640 were followed with a median follow-up time 45.74 months for determining association of long-term major adverse cardiac events (MACE) and genotypes of polymorphisms. There was no significant difference in the frequency of 1565C/T polymorphism between the MI and non-MI groups. The frequency of wild genotype was 69.2% and 72.2%, the frequency of homozygous genotype was 21.3% and 18.4%, and the frequency of mutant genotype was 9.5% and 9.5%, respectively (p=0.505). Results were also similar when adjusted for covariates in a multivariate logistic regression model. No significant difference was also found in total-MACE free survival rate between the patients with different genotypes of 1565C/T polymorphism in both MI and non-MI group. The carriage of the 1565C/T polymorphism of ITGB3 gene seems unlikely to be a significant risk factor for the development of MI in Iranian patients with premature CAD. The presence of this ITGB3 gene polymorphism may not also predict long-term cardiac events.

Keywords: coronary artery disease, myocardial infarction, gene, integrin, beta 3, polymorphism

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Authors: Fulufhelo Amanda Doboro, Kgomotso Sebeko, Stephen Njiro, Moritz Van Vuuren

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Ovine herpesvirus 2 (OvHV-2) genome obtained from the lymphopblastoid cell line of a BJ1035 cow was recently sequenced in the United States of America (USA). Information on the sequences of OvHV-2 genes obtained from South African strains from bovine or other African countries and molecular characterization of OvHV-2 is not documented. Present investigation provides information on the nucleotide and derived amino acid sequences and genetic diversity of Ov 7, Ov 8 ex2, ORF 27 and ORF 73 genes, of these genes from OvHV-2 strains circulating in South Africa. Gene-specific primers were designed and used for PCR of DNA extracted from 42 bovine blood samples that previously tested positive for OvHV-2. The expected PCR products of 495 bp, 253 bp, 890 bp and 1632 bp respectively for Ov 7, Ov 8 ex2, ORF 27 and ORF 73 genes were sequenced and multiple sequence analysis done on the selected regions of the sequenced PCR products. Two genotypes for ORF 27 and ORF 73 gene sequences, and three genotypes for Ov 7 and Ov 8 ex2 gene sequences were identified, and similar groupings for the derived amino acid sequences were obtained for each gene. Nucleotide and amino acid sequence variations that led to the identification of the different genotypes included SNPs, deletions and insertions. Sequence analysis of Ov 7 and ORF 27 genes revealed variations that distinguished between sequences from SA and reference OvHV-2 strains. The implication of geographic origin among SA sequences was difficult to evaluate because of random distribution of genotypes in the different provinces, for each gene. However, socio-economic factors such as migration of people with animals, or transportation of animals for agricultural or business use from one province to another are most likely to be responsible for this observation. The sequence variations observed in this study have no impact on the antibody binding activities of glycoproteins encoded by Ov 7, Ov 8 ex2 and ORF 27 genes, as determined by prediction of the presence of B cell epitopes using BepiPred 1.0. The findings of this study will be used for selection of gene candidates for the development of diagnostic assays and vaccine development as well.

Keywords: amino acid, genetic diversity, genes, nucleotide

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Authors: Philippa Saunders, Claire Fletcher

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INTRODUCTION: Prostate cancer is the most prevalent malignancy affecting Western males. It is initially an androgen-dependent disease: androgens bind to the androgen receptor and drive the expression of genes that promote proliferation and evasion of apoptosis. Despite reduced androgen dependence in advanced prostate cancer, androgen receptor signaling remains a key driver of growth. Androgen deprivation therapy (ADT) is, therefore, a first-line treatment approach and works well initially, but resistance inevitably develops. Abiraterone and Enzalutamide are drugs widely used in ADT and are androgen synthesis and androgen receptor signaling inhibitors, respectively. The shortage of other treatment options means acquired resistance to these drugs is a major clinical problem. MicroRNAs (miRs) are important mediators of post-transcriptional gene regulation and show altered expression in cancer. Several have been linked to the development of resistance to ADT. Manipulation of such miRs may be a pathway to breakthrough treatments for advanced prostate cancer. This study aimed to validate ADT resistance-implicated miRs and their clinically relevant targets. MATERIAL AND METHOD: Small RNA-sequencing of Abiraterone- and Enzalutamide-resistant C42 prostate cancer cells identified subsets of miRs dysregulated as compared to parental cells. Real-Time Quantitative Reverse Transcription PCR (qRT-PCR) was used to validate altered expression of candidate ADT resistance-implicated miRs 195-5p, 497-5p and 29a-5p in ADT-resistant and -responsive prostate cancer cell lines, patient-derived xenografts (PDXs) and primary prostate cancer explants. RESULTS AND DISCUSSION: This study suggests a possible role for miR-497-5p in the development of ADT resistance in prostate cancer. MiR-497-5p expression was increased in ADT-resistant versus ADT-responsive prostate cancer cells. Importantly, miR-497-5p expression was also increased in Enzalutamide-treated, castrated (ADT-mimicking) PDXs versus intact PDXs. MiR-195-5p was also elevated in ADT-resistant versus -responsive prostate cancer cells, while there was a drop in miR-29a-5p expression. Candidate clinically relevant targets of miR-497-5p in prostate cancer were identified by mining AGO-PAR-CLIP-seq data sets and may include AVL9 and FZD6. CONCLUSION: In summary, this study identified microRNAs that are implicated in prostate cancer resistance to androgen deprivation therapy and could represent novel therapeutic targets for advanced disease.

Keywords: microRNA, androgen deprivation therapy, Enzalutamide, abiraterone, patient-derived xenograft

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Authors: Yuriy A. Knirel

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Enteric bacteria Escherichia coli is the predominant facultative anaerobe of the colonic flora, and some specific serotypes are associated with enteritis, hemorrhagic colitis, and hemolytic uremic syndrome. Shigella spp. are human pathogens that cause diarrhea and bacillary dysentery (shigellosis). They are in effect E. coli with a specific mode of pathogenicity. Strains of Salmonella enterica are responsible for a food-borne infection (salmonellosis), and specific serotypes cause typhoid fever and paratyphoid fever. All these bacteria are closely related in respect to structure and genetics of the lipopolysaccharide, including the O-polysaccharide part (O‑antigen). Being exposed to the bacterial cell surface, the O antigen is subject to intense selection by the host immune system and bacteriophages giving rise to diverse O‑antigen forms and providing the basis for typing of bacteria. The O-antigen forms of many bacteria are unique, but some are structurally and genetically related to others. The sequenced O-antigen gene clusters between conserved galF and gnd genes were analyzed taking into account the O-antigen structures established by us and others for all S. enterica and Shigella and most E. coli O-serogroups. Multiple genetic mechanisms of diversification of the O-antigen forms, such as lateral gene transfer and mutations, were elucidated and are summarized in the present paper. They include acquisition or inactivation of genes for sugar synthesis or transfer or recombination of O-antigen gene clusters or their parts. The data obtained contribute to our understanding of the origins of the O‑antigen diversity, shed light on molecular evolutionary relationships between the O-antigens of enteric bacteria, and open a way for studies of the role of gene polymorphism in pathogenicity.

Keywords: enteric bacteria, O-antigen gene cluster, polysaccharide biosynthesis, polysaccharide structure

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Authors: Kumaran Narayanan, Pei-Sheng Liew

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This paper adapts the bacteriophage N15 protelomerase enzyme to assemble linear chromosomes as vectors for gene expression in human cells. Phage N15 has the unique ability to replicate as a linear plasmid with telomeres in E. coli during its prophage stage of life-cycle. The virus-encoded protelomerase enzyme cuts its circular genome and caps its ends to form hairpin telomeres, resulting in a linear human-chromosome-like structure in E. coli. In mammalian cells, however, no enzyme with TelN-like activities has been found. In this work, we show for the first-time transfer of the protelomerase from phage into human and mouse cells and demonstrate recapitulation of its activity in these hosts. The function of this enzyme is assayed by demonstrating cleavage of its target DNA, followed by detecting telomere formation based on its resistance to recBCD enzyme digestion. We show protelomerase expression persists for at least 60 days, which indicates limited silencing of its expression. Next, we show that an intact human β-globin gene delivered on this linear chromosome accurately retains its expression in the human cellular environment for at least 60 hours, demonstrating its stability and potential as a vector. These results demonstrate that the N15 protelomerse is able to function in mammalian cells to cut and heal DNA to create telomeres, which provides a new tool for creating novel structures by DNA resolution in these hosts.

Keywords: chromosome, beta-globin, DNA, gene expression, linear vector

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Authors: Kevser Cakmak, Adrián Montesano, Lourdes Artigas, Marta Salla, Clara Mateu

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The main objective of this research is to explore the relational letter writing technique as an intervention in couple therapy for reconnecting couples with their values and ethical preferences. This is a recently developed therapeutic tool within the framework of Narrative Therapy that consists of two interviews and a letter writing task, in which a meta-conversation between the relationship itself, the couple members, and the therapists is fostered. Although this specific therapeutic technique can be used within the therapy process, in this case study, it is used as a brief stand-alone narrative intervention for a middle age heterosexual couple breast cancer survivor. Couple’s relational and personal wellbeing was monitored before, during, and after the intervention by means of the dyadic adjustment and the clinical outcomes in routine evaluation-outcome measure, respectively. The couple showed a significant improvement after the intervention in both levels. The content of the letter writing exercises was qualitatively analysed to explore the reconstruction of their ethical values. Results from both methods are integrated in order to get an in-depth perspective of the newly developed tool. The potential of the letter writing technique as stand-alone and as adjunct brief intervention is discussed.

Keywords: couple therapy, narrative therapy, psychotherapy tool, relational letter writing

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Authors: Eva Kirzner

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Based on outcome studies published over the past two decades, in 2018, the ACC/AHA published new guidelines for the management of hypercholesterolemia that incorporate the use of coronary artery calcium (CAC) scanning as a decision tool for ascertaining which patients may benefit from statin therapy. This use is based on the recognition that the absence of calcium on CAC scanning (i.e., a CAC score of zero) usually signifies the absence of significant atherosclerotic deposits in the coronary arteries. Specifically, in patients with a high risk for atherosclerotic cardiovascular disease (ASCVD), initiation of statin therapy is generally recommended to decrease ASCVD risk. However, among patients with intermediate ASCVD risk, the need for statin therapy is less certain. However, there is a need for new outcome studies that provide evidence that the management of hypercholesterolemia based on these new ACC/AHA recommendations is safe for patients. Based on a Pub-Med and Google Scholar literature search, four relevant population-based or patient-based cohort studies that studied the relationship between CAC scanning, risk assessment or mortality, and statin therapy that were published between 2017 and 2021 were identified (see references). In each of these studies, patients were assessed for their baseline risk for atherosclerotic cardiovascular disease (ASCVD) using the Pooled Cohorts Equation (PCE), an ACC/AHA calculator for determining patient risk based on assessment of patient age, gender, ethnicity, and coronary artery disease risk factors. The combined findings of these four studies provided concordant evidence that a zero CAC score defines patients who remain at low clinical risk despite the non-use of statin therapy. Thus, these new studies confirm the use of CAC scanning as a safe tool for reducing the potential overuse of statin therapy among patients with zero CAC scores. Incorporating these new data suggest the following best practice: (1) ascertain ASCVD risk according to the PCE in all patients; (2) following an initial attempt trial to lower ASCVD risk with optimal diet among patients with elevated ASCVD risk, initiate statin therapy for patients who have a high ASCVD risk score; (3) if the ASCVD score is intermediate, refer patients for CAC scanning; and (4) and if the CAC score is zero among the intermediate risk ASCVD patients, statin therapy can be safely withheld despite the presence of an elevated serum cholesterol level.

Keywords: cholesterol, cardiovascular disease, statin therapy, coronary calcium

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