Search results for: breast screening

Authors: Faraz Zarghami, Elham Anari, Nosratollah Zarghami, Yones Pilehvar-Soltanahmadi, Abolfazl Akbarzadeh, Sepideh Jalilzadeh-Tabrizi

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The development of nanotherapy has presented a new method of drug delivery targeted directly to the neoplasmic tissues, to maximize the action with fewer dose requirements. In the past two decades, poly(lactic-co-glycolic acid) (PLGA) has frequently been investigated by many researchers and is a popular polymeric candidate, due to its biocompatibility and biodegradability, exhibition of a wide range of erosion times, tunable mechanical properties, and most notably, because it is a FDA-approved polymer. Chrysin is a natural flavonoid which has been reported to have some significant biological effects on the processes of chemical defense, nitrogen fixation, inflammation, and oxidation. However, the low solubility in water decreases its bioavailability and consequently disrupts the biomedical benefits. Being loaded with PLGA-PEG increases chrysin solubility and drug tolerance, and decreases the discordant effects of the drug. The well-structured chrysin efficiently accumulates in the breast cancer cell line (T47D). In the present study, the structure and chrysin loading were delineated using proton nuclear magnetic resonance (HNMR), Fourier-transform infrared spectroscopy (FT-IR), and scanning electron microscopy (SEM), and the in vitro cytotoxicity of pure and nanochrysin was studied by the MTT assay. Next, the RNA was exploited and the cytotoxic effects of chrysin were studied by real-time PCR. In conclusion, the nanochrysin therapy developed is a novel method that could increase cytotoxicity to cancer cells without damaging the normal cells, and would be promising in breast cancer therapy.

Keywords: MTT assay, chrysin, flavonoids, nanotherapy

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Authors: Randa M. T. Mohamed

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Spices have long been used as traditional ingredients in the kitchen for seasoning, coloring, aromatic and food preservative properties. Besides, spices are equally used for therapeutic purposes. The objective of this study was to survey and document the medicinal properties of spices commonly used in the Sudanese kitchen for different food preparations. Also, extracts from reported spices were screened for the presence of secondary metabolites as well as their antioxidant and beta-lactamase inhibitory properties. This study was conducted in the Rekabbya Quartier in Omdurman, Khartoum State, Sudan. Information was collected by carrying out semi-structured interviews. All informants (30) in the present study were women. Spices were purchased from Attareen shop in Omdurman. Essential oils from spices were extracted by hydrodistillation and ethanolic extracts by maceration. Phytochemical screening was performed by thin layer chromatography (TLC). The antioxidant capacity of essential oils and ethanolic extracts was investigated through TLC bioautography. Beta lactamase inhibitory activity was performed by the acidimetric test. Ethnobotany study showed that a total of 16 spices were found to treat 36 ailments belonging to 10 categories. The most frequently claimed medicinal uses were for the digestive system diseases treated by 14 spices and respiratory system diseases treated by 8 spices. Gynaecological problems were treated by 4 spices. Dermatological diseases were cured by 5 spices while infections caused by tapeworms and other microbes causing dysentery were treated by 3 spices. 4 spices were used to treat bad breath, bleeding gum and toothache. Headache, eyes infection, cardiac stimulation and epilepsy were treated by one spice each. Other health problem like fatigue and loss of appetite and low breast milk production were treated by 1, 3 and 2 spices respectively. The majority (69%, 11/16) of spices were exported from different countries like India, China, Indonesia, Ethiopia, Egypt and Nigeria while 31% (5/16) was cultivated in Sudan. Essential oils of all spices were rich in terpenes while ethanolic extracts contained variable classes of secondary metabolites. Both essential oils and ethanolic extracts of all spices exerted considerable antioxidant activity. Only one extract, Syzygium aromaticum, possessed beta lactamase inhibitory activity. In conclusion, this study could contribute in conserving information on traditional medicinal uses of spices in Sudan. Also, the results demonstrated the potential of some of these spices to exert beneficial antimicrobial and antioxidant effect. Detailed phytochemical and biological assays of these spices are recommended.

Keywords: spices, ethnobotany, phytoconstituents, antioxidant, beta lactamase inhibition

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Authors: Zulaika Mohd Khasiran

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The capability of ionic liquids in various applications makes them attracted by many researchers. They have potential to be developed as “green” solvents for gas separation, especially H2S gas. In this work, it is attempted to predict the solubility of hydrogen sulfide (H2S) in ILs by COSMO-RS method. Since H2S is a toxic pollutant, it is difficult to work on it in the laboratory, therefore an appropriate model will be necessary in prior work. The COSMO-RS method is implemented to predict the Henry’s law constants and activity coefficient of H2S in 140 ILs with various combinations of cations and anions. It is found by the screening that more H2S can be absorbed in ILs with [Cl] and [Ac] anion. The solubility of H2S in ILs with different alkyl chain at the cations not much affected and with different type of cations are slightly influence H2S capture capacities. Even though the cations do not affect much in solubility of H2S, we still need to consider the effectiveness of cation in different way. The prediction results only show their physical absorption ability, but the absorption of H2S need to be consider chemically to get high capacity of absorption of H2S.

Keywords: H2S, hydrogen sulfide, ionic liquids, COSMO-RS

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Authors: Zahra Shokrolahi, Mohammad Reza Atashzar

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The goals of treating patients with breast cancer are to cure the disease, prolong survival, and improve quality of life. Immune cells in the tumor microenvironment have an important role in regulating tumor progression. The term of cellular immunotherapy refers to the administration of living cells to a patient; this type of immunotherapy can be active, such as a dendritic cell (DC) vaccine, in that the cells can stimulate an anti-tumour response in the patient, or the therapy can be passive, whereby the cells have intrinsic anti-tumour activity; this is known as adoptive cell transfer (ACT) and includes the use of autologous or allogeneic lymphocytes that may, or may not, be modified. The most important breast cancer cellular immunotherapies involving the use of T cells and natural killer (NK) cells in adoptive cell transfer, as well as dendritic cells vaccines. T cell-based therapies including tumour-infiltrating lymphocytes (TILs), engineered TCR-T cells, chimeric antigen receptor (CAR T cell), Gamma-delta (γδ) T cells, natural killer T (NKT) cells. NK cell-based therapies including lymphokine-activated killers (LAK), cytokine-induced killer (CIK) cells, CAR-NK cells. Adoptive cell therapy has some advantages and disadvantages some. TILs cell strictly directed against tumor-specific antigens but are inactive against tumor changes due to immunoediting. CIK cell have MHC-independent cytotoxic effect and also need concurrent high dose IL-2 administration. CAR T cell are MHC-independent; overcome tumor MHC molecule downregulation; potent in recognizing any cell surface antigen (protein, carbohydrate or glycolipid); applicable to a broad range of patients and T cell populations; production of large numbers of tumor-specific cells in a moderately short period of time. Meanwhile CAR T cells capable of targeting only cell surface antigens; lethal toxicity due to cytokine storm reported. Here we present the most popular cancer cellular immunotherapy approaches and discuss their clinical relevance referring to data acquired from clinical trials .To date, clinical experience and efficacy suggest that combining more than one immunotherapy interventions, in conjunction with other treatment options like chemotherapy, radiotherapy and targeted or epigenetic therapy, should guide the way to cancer cure.

Keywords: breast cancer , cell therapy , CAR T cell , CIK cells

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Authors: Cassey Younghans

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Background: Current health care trends demonstrate that there is an increasing number of patients being hospitalized with complex comorbidities. These complex needs require advanced therapies, and treatment goals often focus on doing everything possible to prolong life rather than focusing on the individual patient’s quality of life which is the goal of palliative care efforts. Patients benefit from palliative care in the early stages of the illness rather than after the disease progressed or the state of acuity has advanced. The clinical problem identified was that palliative care was not being implemented early enough in the disease process with patients who had complex medical conditions and who would benefit from the philosophy and skills of palliative care professionals. Purpose: The purpose of this quality improvement study was to increase the number of palliative care screenings and consults completed on adults after being admitted to one Non-ICU and Non-COVID hospital unit. Methods: A retrospective chart review assessing for possible missed opportunities to introduce palliation was performed for patients with six primary diagnoses, including heart failure, liver failure, end stage renal disease, chronic obstructive pulmonary disease, cerebrovascular accident, and cancer in a population of adults over the ago of 19 on one medical-surgical unit over a three-month period prior to the intervention. An educational session with the nurses on the benefits of palliative care was conducted by the researcher, and a screening tool was implemented. The expected outcome was to have an increase in early palliative care consultation with patients with complex comorbid conditions and a decrease in missed opportunities for the implementation of palliative care. Another retrospective chart review was completed following completion of the three month piloting of the tool. Results: During the retrospective chart review, 46 patients were admitted to the medical-surgical floor with the primary diagnoses identified in the inclusion criteria. Six patients had palliative care consults completed during that time. Twenty-two palliative care screening tools were completed during the intervention period. Of those, 15 of the patients scored a 7 or higher, suggesting that a palliative care consultation was warranted. The final retrospective chart review identified that 4 palliative consults were implemented during that time of the 31 patients who were admitted over the three month time frame. Conclusion: Educating nurses and implementing a palliative care screening upon admission can be of great value in providing early identification of patients who might benefit from palliative care. Recommendations – It is recommended that this screening tool should be used to help identify the patents of whom would benefit from a palliative care consult, and nurses would be able to initiated a palliative care consultation themselves.

Keywords: palliative care, screening, early, palliative care consult

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Authors: Yuping Liu, Li Tao, Yin Lu

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Accumulating evidence has been shown that diallyl trisulfide (DATS) from garlic may reduce the risk of developing several types of cancer. In view of the dynamic crosstalk interplayed by tumor cells and platelets in hematogenous metastasis, we demonstrate the effectiveness of DATS on the metastatic behaviors of MDA-MB-435s human breast cancer cell line co-incubated with activated platelets. Indeed, our data identified that DATS significantly blocked platelets fouction induced by PAF, followed by the decreased production of TXB2. DATS was found to dose-dependently suppressed MDA-MB-435s cell migration and invasion in presence of activated platelets by PAF in vitro. Furthermore, the expression, secretion and enzymatic activity of matrix metalloproteinase (MMP)-2/9, as well as the luciferase activity of upstream regulator NF-κB in MDA-MB-435s, were obviously diminished by DATS. In parallel, DATS blocked upstream NF-κB activation signaling complexes composed of extracellular signal-related kinase (ERK) as assessed by measuring the levels of the phosphorylated forms.

Keywords: DATS, ERK, metastasis, MMPs, NF-κB, platelet

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Authors: Elida Tafupenji Kamanga

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Cancer of the cervix remains one of the high causes of death among Malawian women. Despite the government introduction of free screening services throughout the country, patronage still remains low and lack of knowledge high. Given the critical role mass media plays in relaying different information to the public including health and its influence on health behaviours, the study sought to analyse Malawi media coverage of the disease and its effectiveness. The findings of the study will help inform media advocacy directed at changing any coverage impeding the effective dissemination of cervical cancer message which consequently will help increase awareness and accessing of screening behaviours among women. A content analysis of 29 newspapers and promotional messages on cervical from a local radio station was conducted for the period from 2012 to 2015. Overall the results showed media coverage in terms of content and frequency increased for the four-year period. However, of concern was the quality of information both media presented to the public. The lapse in information provided means there is little education taking place through the media which could be contributing to the knowledge gap the women have thereby affecting their decision to screen. Also lack of adequate funding to media institutions and lack of collaboration between media institutions and stakeholders involved in the fight against the disease were noted as other contributing factors to low coverage of the disease. Designing messages that are not only informative and educative but also innovative may help increase awareness; improve the knowledge gap and potential adoption of preventive screening behaviour by Malawian women. Conversely, good communication between the media institutions and researchers involved in the fight against the disease through the channelling of new findings back to the public as well as increasing funding towards similar cause should be considered.

Keywords: cervical cancer, effectiveness, media coverage, screening

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Authors: Rashidah Ghazali, Herlina Abdul Rahim, Mashitah Shikh Maidin, Shafishuhaza Sahlan, Noramli Abdul Razak

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Sound exposure effects have been investigated by broadcasting a group of broilers with sound of Quran verses (Group B) whereas the other group is the control broilers (Group C). The growth rate comparisons in terms of weight and raw meat texture measured by shear force have been investigated. Twenty-seven broilers were randomly selected from each group on Day 24 and weight measurement was carried out every week till the harvest day (Day 39). Group B showed a higher mean weight on Day 24 (1.441±0.013 kg) than Group C. Significant difference in the weight on Day 39 existed for Group B compared to Group C (p< 0.05). However, there was no significant (p> 0.05) difference of shear force in the same muscles (breast and drumstick raw meat) of both groups but the shear force of the breast meat for Group B and C broilers was lower (p < 0.05) than that of their drumstick meat. Thus, broadcasting the sound of Quran verses in the coop can be applied to improve the growth rate of broilers for producing better quality poultry.

Keywords: broilers, sound, shear force, weight

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Authors: Soroosh Torabi, Brad Berron, Ren Xu, Christine Trinkle

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Microfluidics integrated with 3D cell culture is a powerful technology to mimic cellular environment, and can be used to study cell activities such as proliferation, migration and response to drugs. This technology has gained more attention in cancer studies over the past years, and many organ-on-a-chip systems have been developed to study cancer cell behaviors in an ex-vivo tumor microenvironment. However, there are still some barriers to adoption which include low throughput, complexity in 3D cell culture integration and limitations on non-optical analysis of cells. In this study, a user-friendly microfluidic multi-well plate was developed to mimic the in vivo tumor microenvironment. The microfluidic platform feeds multiple 3D cell culture sites at the same time which enhances the throughput of the system. The platform uses hydrophobic Cassie-Baxter surfaces created by microchannels to enable convenient loading of hydrogel/cell suspensions into the device, while providing barrier free placement of the hydrogel and cells adjacent to the fluidic path. The microchannels support convective flow and diffusion of nutrients to the cells and a removable lid is used to enable further chemical and physiological analysis on the cells. Different breast cancer cell lines were cultured in the device and then monitored to characterize nutrient delivery to the cells as well as cell invasion and proliferation. In addition, the drug response of breast cancer cell lines cultured in the device was compared to the response in xenograft models to the same drugs to analyze relevance of this platform for use in future drug-response studies.

Keywords: microfluidics, multi-well 3d cell culture, tumor microenvironment, tumor-on-a-chip

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Authors: Cristina Fernandez, Felix Amparano, John Tumberger, Stephani Stancil, Sarah Hampl, Brooke Sweeney, Amy R. Beck, Helena H Laroche, Jared Tucker, Eileen Chaves, Sara Gould, Matthew Lindquist, Lora Edwards, Renee Arensberg, Meredith Dreyer, Jazmine Cedeno, Alleen Cummins, Jennifer Lisondra, Katie Cox, Kelsey Dean, Rachel Perera, Nicholas A. Clark

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Background: Adolescents with obesity are at higher risk of disordered eating than the general population. Detection of eating disorders (ED) is difficult. Screening questionnaires may aid in early detection of ED. Our team’s prior efforts focused on increasing ED screening rates to ≥90% using a validated 10-question adolescent binge eating disorder screening questionnaire (ADO-BED). This aim was achieved. We then aimed to improve treatment plan initiation of patients ≥12 years of age who screen positive for BED within our WMC from 33% to 70% within 12 months. Methods: Our WMC is within a tertiary-care, free-standing children’s hospital. A3, an improvement framework, was used. A multidisciplinary team (physicians, nurses, registered dietitians, psychologists, and exercise physiologists) was created. The outcome measure was documentation of treatment plan initiation of those who screen positive (goal 70%). The process measure was ADO-BED screening rate of WMC patients (goal ≥90%). Plan-Do-Study-Act (PDSA) cycle 1 included provider education on current literature and treatment plan initiation based upon ADO-BED responses. PDSA 2 involved increasing documentation of treatment plan and retrain process to providers. Pre-defined treatment plans were: 1) repeat screen in 3-6 months, 2) resources provided only, or 3) comprehensive multidisciplinary weight management team evaluation. Run charts monitored impact over time. Results: Within 9 months, 166 patients were seen in WMC. Process measure showed sustained performance above goal (mean 98%). Outcome measure showed special cause improvement from mean of 33% to 100% (n=31). Of treatment plans provided, 45% received Plan 1, 4% Plan 2, and 46% Plan 3. Conclusion: Through a multidisciplinary improvement team approach, we maintained sustained ADO-BED screening performance, and, prior to our 12-month timeline, achieved our project aim. Our efforts may serve as a model for other multidisciplinary WMCs. Next steps may include expanding project scope to other WM programs.

Keywords: obesity, pediatrics, clinic, eating disorder

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Authors: Nihar Ranjan, Derrick Watkins, Dev P. Arya

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Current methods in the study of antibiotic activity of ribosome targeted antibiotics are dependent on cell based bacterial inhibition assays or various forms of ribosomal binding assays. These assays are typically independent of each other and little direct correlation between the ribosomal binding and bacterial inhibition is established with the complementary assay. We have developed novel high-throughput capable assays for ribosome targeted drug discovery. One such assay examines the compounds ability to bind to a model ribosomal RNA A-site. We have also coupled this assay to other functional orthogonal assays. Such analysis can provide valuable understanding of the relationships between two complementary drug screening methods and could be used as standard analysis to correlate the affinity of a compound for its target and the effect the compound has on a cell.

Keywords: bacterial resistance, aminoglycosides, screening, drugs

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Authors: Marta E. Hernandez-Caballero, Veronica Borgonio-Cuadra, Antonio Miranda-Duarte, Xochitl Rojas-Toledo, Normand Garcia-Hernandez, Maura Cardenas-Garcia, Teresa Abad-Camacho

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Breast cancer (BC) is the most common tumor in women over the world. DNA methylation is an epigenetic modification critical in CpG sites, aberrant methylation of CpG islands in promoters is a hallmark of cancer. So, gene expression can be regulated by alterations in DNA methylation. In cell lines DACT2 gene reduces the growth and migration of tumor cells by its participation in the suppression of TGFb/SMAD2/3. PHF20L1 is involved in histone acetylation therefore, it regulates transcription. Our aim was to analyze the methylation status of the DACT2 and PHF20L1 promoter regions in tumoral and healthy mammary tissue from women with BC in different progression states. The study included 77 patients from Centro Medico Nacional La Raza in Mexico City. After identifying a CpG island in DACT2 and PHF20L1 promoters, DNA methylation status was analyzed through sodium bisulfite with subsequent amplification using methylation-specific PCR. Results revealed no changes in methylation status of PHF20L1 and cancer stages (II y III) or in comparison to healthy tissues, it was demethylated. DACT2 promoter methylation was no significant between tumoral stages (II, P = 0.37; III, P = 0.17) or with healthy tissue. Previous data reported DACT2 methylated in nasopharyngeal carcinoma but in this study promoter methylation was not observed. PHF20L1 protein contains N-terminal Tudor and C-terminal plant homeodomain domains, it has been suggested that can stabilize DNMT1 regulating DNA methylation, therefore, was associated with poor prognostic in BC. We found no evidence of methylation in patients and controls in PHF20L1 promoter, so its association with BC may have no direct relation with promoter methylation. More studies including other methylation sites in these genes in BC are necessary.

Keywords: bisulfite conversion, breast cancer, DACT2, DNA methylation, PHF20L1, tumoral status

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Authors: Kabange Kasumbwe, Viresh Mohanlall, Bharti Odhav, Venu Narayanaswamy

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Coumarins are naturally occurring plant metabolites known for their pharmacological properties such as anticoagulant, antimicrobial, anticancer, antioxidant, anti-inflammatory and antiviral properties. The pharmacological and biochemical properties and curative applications of coumarins depend on the substitution around the coumarin core structure. In the present study, seven halogenated coumarins CMRN1-CMRN7 were synthesized and evaluated for their anticancer activity. The cytotoxicity potential of the test compounds was evaluated against UACC62 (Melanoma), MCF-7 (Breast cancer) and PBM (Peripheral Blood Mononuclear) cell lines using MTT assay keeping doxorubicin as standard drug. The apoptotic potential of the coumarin compounds was evaluated against UACC62 (Melanoma) cell by assessing their morphological changes, membrane change, mitochondria membrane potential; pro-apoptotic changes were investigated using the AnnexinV-PI staining, JC-1, caspase-3 enzyme kits respectively on flow cytometer. The synthetic coumarin has strongly suppressed the cell proliferation of UACC-62 (Melanoma) and MCF-7 (Breast) Cancer cells, the higher toxicity of these compounds against UACC-62 (Melanoma) and MCF-7 (Breast) were CMRN3, CMRN4, CMRN5, CMRN6. However, compounds CMRN1, CMRN2, and CMRN7 had no significant inhibitory effect. Furthermore the active compounds CMRN3, CMRN4, CMRN5, CMRN6 exerted antiproliferative effects through apoptosis induction against UACC-62 (Melanoma), suggesting their potential could be considered as attractive lead molecules in the future for the development of potential anticancer agents since one of the important criteria in the development of therapeutic drugs for cancer treatment is to have high selectivity and less or no side-effects on normal cells and these compounds had no inhibitory effect against the PBMC cells.

Keywords: coumarin, MTT, apoptosis, cytotoxicity

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Authors: Wahid Terro, Franco Lugnani, Miray Terro

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Roughly 90% of mouth cancer is squamous cell carcinoma. Oral region metastases are considered uncommon. The most frequent sources of metastatic tumors to the mouth are primary cancers from the lung and breast, with the latter being the most common site for tumors that metastasize to the jawbones. Treatment involves surgery, radiation, chemotherapy or a combination of these. However, these treatment modalities entail severe aesthetic and functional side effects. The aim of this article is to present an alternative and unprecedented treatment modality for metastatic mandibular bone carcinoma – Cryosurgery. A 37-year-old female patient presented with gradually increasing pain over several weeks whose radiological tests revealed a formation of mass in the left mandibular ramus. A biopsy obtained by bone puncture of the ramus was reported to be metastatic breast squamous cell carcinoma. The patient has been successfully treated without esthetic and functional side effects by using cryosurgery. We conclude that it is value promoting this method for current and future use.

Keywords: squamous cell carcinoma, mandible, cryosurgery, metastasis

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Authors: Jelena Nedeljković Trailović, Marko Vasiljević, Jog Raj, Hunor Farkaš, Branko Petrujkić, Stamen Radulović, Gorana Popvić

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The experiment was performed on a total of 99 one-day-old broilers of Cob 500 provenance, which were divided into IX equal groups. Broilers of the E-I group were fed 0.25 mg T-2 toxin/kg feed, E-II and E-III groups 0.25 mg T-2 toxin/kg feed with the addition of 1 kg/t and 3 kg/t of the mycotoxin detoxification agent MDA, respectively. The E-IV group received 1 mg of T-2 toxin/kg of feed, and the broilers of E-V and E-VI groups received 1 mg of T-2 toxin/kg of feed with the addition of 1 kg/t and 3 kg/t of the MDA detoxification preparation, respectively. The E-VII group received commercial feed without toxins and additives, the E-VIII and E-IX groups received feed with 1kg/t and 3kg/t of the MDA detoxification preparation. The trial lasted 42 days. Observing the results obtained on the 42nd day of the experiment, we can conclude that the change in the absolute mass of the spleen occurred in the broilers of the E-IV group (1.66±0.14)g, which was statistically significantly lower compared to the broilers of the E-V and E-VI groups (2.58±0.15 and 2.68±0.23)g. Heart mass was significantly statistically lower in broilers of group E-IV (9.1±0.38)g compared to broilers of group E-V and E-VI (12.23±0.5 and 11.43±0.51)g. It can be concluded that the broilers that received 1 kg/t and 3 kg/t of the detoxification preparation had an absolute mass of organs within physiological limits. Broilers of the E-IV group achieved the lowest BM during the experiment (on the 42nd day of the experiment 1879±52.73)g, they were significantly statistically lower than the BW of broilers of all experimental groups. This trend is observed from the beginning to the end of the experiment. The protective effect of the detoxification preparation can be seen in broilers of the E-V group, that had a significantly statistically higher BM on the 42nd day of the experiment (2225±58.81)g compared to broilers of group E-IV. Broilers of E-VIII group (2452±46.71) g, which received commercial feed with the addition of 1 kg/t MDA preparation, had the highest BMI at the end of the experiment. At the end of the trial on the 42nd day, blood samples were collected from broilers of the experimental groups that received T-2 toxin and MR detoxification preparations in different concentrations. Also, liver and breast musculature samples were collected for testing for the presence and content of T-2 toxin, HT-2 toxin, T-2 tetraol and T-2 triol. Due to very rapid elimination from the blood, no remains of T-2 toxin and its metabolites were detected in the blood of broilers of groups E-I to E-VI. In the breast muscles, T-2 toxin residues below LoQ < 0.2 (μg/kg) were detected in all groups that received T-2 toxin in food, the highest value was recorded in the E-IV group (0.122 μg/kg and the lowest in E -VI group 0.096 μg/kg). No T-2 toxin residues were detected in the liver. Remains of HT-2 were detected in the breast muscles and livers of broilers from E-IV, E-V and E-VI groups, LoQ < 1 (μg/kg); for the breast muscles: 0.054, 0.044 and 0.041 μg/kg, and for the liver: 0.473, 0.231 and 0.185 μg/kg. Summing up all the results, a partial protective effect of the detoxification preparation, added to food in the amount of 1kg/t, can be seen.

Keywords: T-2 toxin, bloiler, MDA, mycotoxuns

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Authors: Tamar Grdzelishvili, Zaza Sinauridze

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Introduction: Preeclampsia is a complication of the second trimester of pregnancy, which is characterized by high morbidity and multiorgan damage. Many complex pathogenic mechanisms are now implicated to be responsible for this disease (1). Preeclampsia is one of the leading causes of maternal mortality worldwide. Statistics are enough to convince you of the seriousness of this pathology: about 100,000 women die of preeclampsia every year. It occurs in 3-14% (varies significantly depending on racial origin or ethnicity and geographical region) of pregnant women, in 75% of cases - in a mild form, and in 25% - in a severe form. During severe pre-eclampsia-eclampsia, perinatal mortality increases by 5 times and stillbirth by 9.6 times. Considering that the only way to treat the disease is to end the pregnancy, the main thing is timely diagnosis and prevention of the disease. Identification of high-risk pregnant women for PE and giving prophylaxis would reduce the incidence of preterm PE. First-trimester screening model developed by the Fetal Medicine Foundation (FMF), which uses the Bayes-theorem to combine maternal characteristics and medical history together with measurements of mean arterial pressure, uterine artery pulsatility index, and serum placental growth factor, has been proven to be effective and have superior screening performance to that of traditional risk factor-based approach for the prediction of PE (2) Methods: Retrospective single center screening study. The study population consisted of women from the Tbilisi maternity hospital “Pineo medical ecosystem” who met the following criteria: they spoke Georgian, English, or Russian and agreed to participate in the study after discussing informed consent and answering questions. Prior to the study, the informed consent forms approved by the Institutional Review Board were obtained from the study subjects. Early assessment of preeclampsia was performed between 11-13 weeks of pregnancy. The following were evaluated: anamnesis, dopplerography of the uterine artery, mean arterial blood pressure, and biochemical parameter: Pregnancy-associated plasma protein A (PAPP-A). Individual risk assessment was performed with performed by Fast Screen 3.0 software ThermoFisher scientific. Results: A total of 513 women were recruited and through the study, 51 women were diagnosed with preeclampsia (34.5% in the pregnant women with high risk, 6.5% in the pregnant women with low risk; P<0.000 1). Conclusions: First-trimester screening combining maternal factors with uterine artery Doppler, blood pressure, and pregnancy-associated plasma protein-A is useful to predict PE in a routine care setting. More patient studies are needed for final conclusions. The research is still ongoing.

Keywords: first-trimester, preeclampsia, screening, pregnancy-associated plasma protein

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Authors: Ibtissem El Ouar, Cornelia Braicu, Dalila Naimi, Alexendru Irimie, Ioana Berindan-Neagoe

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Helix aspersa, 'the garden snail' is a big land snail widely found in the Mediterranean countries, it is one of the most consumed species in the west of Algeria. It is commonly used in zootherapy to purify blood and to treat cardiovascular diseases and liver problems. The aim of our study is to investigate, the antitumor activity of an aqueous extract from Helix aspersa prepared by the traditional method on Hs578T; a triple negative breast cancer cell line. Firstly, the free radical scavenging activity of H. aspersa extract was assessed by measuring its capability for scavenging the radical 2,2-diphenyl-1-picrylhydrazyl (DPPH), as well as its ability to reduce ferric ion by the FRAP assay (ferric reducing ability). The cytotoxic effect of H. aspersa extract against Hs578T cells was evaluated by the MTT test (3-(4,5- dimethylthiazl-2-yl)-2,5- diphenyltetrazolium bromide)) while the mode of cell death induced by the extract has been determined by fluorescence microscopy using acredine orange/ethidium bromide (AO/EB) probe. The level of TNFα has also measured in cell medium by ELISA method. The results suggest that H. aspersa extract has an antioxidant activity, especially at high concentrations, it can reduce DPPH radical and ferric ion. The MTT test shows that H. aspersa extract has a great cytotoxic effect against breast cancer cells, the IC50 value correspond of the dilution 1% of the crude extract. Moreover, the AO/EB staining shows that TNFα induced necrosis is the main form of cell death induced by the extract. In conclusion, the present study may open new perspectives in the search for new natural anticancer drugs.

Keywords: breast cancer, Helix aspersa, Hs578t cell line, necrosis

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Authors: M. Marjaneh, M. Y. Narazah, H. Shahrul

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Array-based gene expression analysis is a powerful tool to profile expression of genes and to generate information on therapeutic effects of new anti-cancer compounds. Anti-apoptotic effect of thymoquinone was studied in MCF7 breast cancer cell line using gene expression profiling with cDNA micro array. The purity and yield of RNA samples were determined using RNeasyPlus Mini kit. The Agilent RNA 6000 Nano LabChip kit evaluated the quantity of the RNA samples. AffinityScript RT oligo-dT promoter primer was used to generate cDNA strands. T7 RNA polymerase was used to convert cDNA to cRNA. The cRNA samples and human universal reference RNA were labelled with Cy-3-CTP and Cy-5-CTP, respectively. Feature Extraction and GeneSpring software analysed the data. The single experiment analysis revealed involvement of 64 pathways with up-regulated genes and 78 pathways with down-regulated genes. The MAPK and p38-MAPK pathways were inhibited due to the up-regulation of PTPRR gene. The inhibition of p38-MAPK suggested up-regulation of TGF-ß pathway. Inhibition of p38 - MAPK caused up-regulation of TP53 and down-regulation of Bcl2 genes indicating involvement of intrinsic apoptotic pathway. Down-regulation of CARD16 gene as an adaptor molecule regulated CASP1 and suggested necrosis-like programmed cell death and involvement of caspase in apoptosis. Furthermore, down-regulation of GPCR, EGF-EGFR signalling pathways suggested reduction of ER. Involvement of AhR pathway which control cytochrome P450 and glucuronidation pathways showed metabolism of Thymoquinone. The findings showed differential expression of several genes in apoptosis pathways with thymoquinone treatment in estrogen receptor-positive breast cancer cells.

Keywords: cDNA microarray, thymoquinone, CARD16, PTPRR, CASP10

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Authors: Maksim Barsakov

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Introduction. In modern plastic surgery, there are a large number of breast lift techniques.Due to the spreading information about the "side effects" of silicone implants, interest in implant-free mastopexy is increasing year after year. However, despite the variety of techniques, patients sometimes do not get full satisfaction from the results of mastopexy because of the unexpressed filling of the upper pole, extended anchoring postoperative scars and sometimes because of obtaining an aesthetically unattractive breast shape. The stability of the result after mastopexy depends on many factors, including postoperative rehabilitation. Stability of weight and hormonal background, stretchability of tissues. The high recurrence rate of ptosis and short-term aesthetic effect of mastopexy indicate the urgency of improving surgical techniques and increasing the stabilization of breast tissue. Purpose of the study. To develop and introduce into practice a technique of mastopexy based on the use of a modified Ribeiro flap, as well as elements of tissue movement and fixation designed to increase the stability of postoperative mastopexy. In addition, to give indications for the application of this surgical technique. Materials and Methods. it operated on 103 patients aged 18 to 53 years from 2019 to 2023 according to the reported method. These were patients with primary mastopexy, secondary mastopexy, and also patient with implant removal and one-stage mastopexy. The patients were followed up for 12 months to assess the stability of the result. Results and their discussion. Observing the patients, we noted greater stability of the breast shape and upper pole filling compared to the conventional classical methods. We did not have to resort to anchoring scars. In 90 percent of cases, a inverted T-shape scar was used. In 10 percent, the J-scar was used. The quantitative distribution of complications identified among the operated patients is as follows: worsened healing of the junction of vertical and horizontal sutures at the period of 1-1.5 months after surgery - 15 patients; at treatment with ointment method healing was observed in 7-30 days; permanent loss of NAC sensitivity - 0 patients; vascular disorders in the area of NAC/areola necrosis - 0 patients; marginal necrosis of the areola-2 patients. independent healing within 3-4 weeks without aesthetic defects. Aesthetically unacceptable mature scars-3 patients; partial liponecrosis of the autoflap unilaterally - 1 patient. recurrence of ptosis - 1 patient (after weight loss of 12 kg). In the late postoperative period, 2 patients became pregnant, gave birth, and no lactation problems were observed. Conclusion. Thus, in the world of plastic surgery methods of breast lift continue to improve, which is especially relevant in modern times, due to the increased attention to this operation. The author's proposed method of mastopexy with glandular autoflap allows obtaining in most cases a stable result, a fuller breast shape, avoiding the presence of extended anchoring scars, and also preserves the possibility of lactation. The author of this article has obtained a patent for invention for this method of mastopexy.

Keywords: mastopexy, mammoplasty, autoflap, personal technique

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Authors: Manisha Singh, Padam Simkhada

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Background: Although mental health policy has been adapted in Nepal since 1997, the implementation of the policy framework is yet to happen. The fact that mental health services are largely concentrated in urban areas more specific to treatment only provides a clear picture of the scarcity of mental health services in the country. The shreds of evidence from around the world, along with WHO’s (World Health Organization) Mental Health Gap Action Program (mhGAP) suggest that effective mental health services can be provided from Primary Health Care (PHC) centers through community-based programs without having to place a specialized health worker. However, the country is still facing the same challenges to date with very few psychiatrists and psychologists, but they are largely based in cities. Objectives: The main objectives of this study are; (a) to understand the perception of health workers at PHC on maternal mental health, and (b) to assess the availability of the mental health services at PHC to address maternal mental health. Methods: This study used a qualitative approach where an in-depth interview was conducted with the health workers at the primary level. “Mayadevi” rural municipality in Rupendehi District that comprised of 13 small villages, was chosen as the study site. A total 8 health institutions which covered all 13 sites were included where either the health post in- charge or health worker working in maternal and child health care was interviewed for the study. All the health posts in the study area were included in the study. The interviews were conducted in Nepali; later, they were translated in English, transcribed, and triangulated. NViVO was used for the analysis. Results: The findings show that most of the health workers understood what maternal mental health was and deemed it as a public health issue. They could explain the symptoms and knew what medication to prescribe if need be. However, the majority of them failed to name the screening tools in place for maternal mental health. Moreover, they hadn’t even seen one. None of the health care centers had any provision for screening mental health status. However, one of the centers prescribed medication when the patients displayed symptoms of depression. But they believed there were a significant number of hidden cases in the community due to the stigma around mental health and being a woman with mental health problem makes the situation even difficult. Nonetheless, the health workers understood the importance of having screening tools and acknowledged the need of training and support in order to provide the services from PHC. Conclusion: Community health workers can identify cases with mental health problems and prevent them from deteriorating further. But there is a need for robust training and support to build the capacity of the health workers. The screening tools on mental health needs to be encouraged to be used in the PHC levels. Furthermore, community-based culture-sensitive programs need to be initiated and implemented to mitigate the stigma related issues around mental health.

Keywords: maternal mental health, health care providers, screening, Nepal

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Authors: Aref Zribi, Sonia Ben Nasr, Sana Fendri, Mahdi Balti, Abderazzek Haddaoui

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Background: Despite their benefit, Endocrine therapies (ET) are known to have substantial adverse events (AEs) such as hot flashes, mood disorders and osteoarticular pain. ET induced alopecia(EIA) is less frequently noted by patients and is less reported in the literature. The aim of our study was to report ET alopecia characteristics and their influence on patient and treatment observance. Method: We conducted a retrospective study including luminal BC patients treated in the oncology department of the military hospital of Tunis between January 2015 and December 2020. Patients treated with previous chemotherapy-inducing alopecia were excluded. Results: 145 female patients were included. The median age was 59 years. EIA was reported in 44% of cases. Alopecia was attributed to aromatase inhibitors in 53% and tamoxifen in 21%. Severity was grade 1 in 80% and grade 2 in the remaining cases. ET discontinuation because of alopecia was noted in 6.5 % of patients. Moderate improvement of alopecia was observed with topical minoxidil and Thallium metallicum 9CH homeopathy during ET in 60% of patients. Conclusions: EIA is frequent in BC patients and should be considered to improve treatment observance and patients’ quality of life.

Keywords: endocrine therapy, alopecia, breast cancer, Tunisia

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Authors: Elaine Wong

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This research protocol aims to explore and justify the need for nutrition and physical activity intervention to improve health outcomes among SME (Small Medium Enterprise) employees. It was found that the worksite is an ideal and convenient setting for employees to take charge of their health thru active participation in health programmes since they spent a great deal of time at their workplace. This study will examine the impact of both general or/and targeted health interventions in both SME and non-SME companies utilizing the Workplace Health Promotion (WHP) grant over a 12 months period and assessed the improvement in chronic health disease outcomes in Singapore. Random sampling of both non-SME and SME companies will be conducted to undergo health intervention and statistical packages such as Statistical Package for Social Science (SPSS) 25 will be used to examine the impact of both general and targeted interventions on employees who participate and those who do not participate in the intervention and their effects on blood glucose (BG), blood lipid, blood pressure (BP), body mass index (BMI), and body fat percentage. Using focus groups and interviews, the data results will be transcribed to investigate enablers and barriers to workplace health intervention revealed by employees and WHP coordinators that could explain the variation in the health screening results across the organisations. Dietary habits and physical activity levels of the employees participating and not participating in the intervention will be collected before and after intervention to assess any changes in their lifestyle practices. It makes economic sense to study the impact of these interventions on health screening outcomes across various organizations that are existing grant recipients to justify the sustainability of these programmes by the local government. Healthcare policy makers and employers can then tailor appropriate and relevant programmes to manage these escalating chronic health disease conditions which is integral to the competitiveness and productivity of the nation’s workforce.

Keywords: chronic diseases, health screening, nutrition and fitness intervention , workplace health

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Authors: Magdalena Twaruzek, Karolina Ropejko

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Mycotoxins are secondary metabolites of molds. Ochratoxin A (OTA) is the most common in the Polish climate. It is produced by fungi of the genera Aspergillus and Penicillium. It is produced as a result of improper food storage. It is present in many products that are consumed both by humans and animals: cereals, wheat gluten, coffee, dried fruit, wine, grape juice, spices, beer, and products based on them. OTA is nephrotoxic, hepatotoxic, potentially carcinogenic, and teratogenic. OTA mainly enters an organism by oral intake. The aim of the study was to detect the presence of OTA in milk, urine, and serum of lactating women. A survey was also conducted regarding the daily diet of women. The research group consisted of 32 lactating women (11 were the donors from the Milk Bank in Toruń, the other 21 were recruited for this study). Results of the analysis showed the occurrence of OTA only in 3 milk samples (9.38%). The minimum level was 0.01 ng/ml, while the maximum 0.018 ng/ml and the mean 0.0013 ng/ml. Twenty-six urine samples (81.25%) were OTA positive, with minimum level 0.013 ng/ml, maximum level 0.117 ng/ml and mean 0.0192 ng/ml. Also, all 32 serum samples (100%) were contaminated by OTA, with a minimum level of 0.099 ng/ml, a maximum level of 2.38 ng/ml, and a mean of 0.4649 ng/ml. In the case of 3 women, OTA was present in all tested body fluids. Based on the results, the following conclusions can be drawn: the breast-milk of women in the study group is slightly contaminated with ochratoxin A. Ten samples of urine contained ochratoxin A above its average content in tested samples. Moreover, serum of 8 women contains ochratoxin A at a level above the average content of this mycotoxin in tested samples. The average ochratoxin A level in serum in the presented studies was 0.4649 ng/ml, which is much lower than the average serum ochratoxin A level established in several countries in the world, i.e., 0.7 ng/ml. Acknowledgment: This study was supported by the Polish Minister of Science and Higher Education under the program 'Regional Initiative of Excellence' in 2019 - 2022 (Grant No. 008/RID/2018/19).

Keywords: breast-milk, urine, serum, contamination, ochratoxin A

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Authors: Junie B. Billones, Maria Constancia O. Carrillo, Voltaire G. Organo, Stephani Joy Y. Macalino, Inno A. Emnacen, Jamie Bernadette A. Sy

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One of the major interferences in the Philippines’ tuberculosis control program is the widespread prevalence of Mtb strains that are resistant to known drugs, such as the MDR-TB (Multi Drug Resistant Tuberculosis) and XDR-TB (Extensively Drug Resistant Tuberculosis). Therefore, there is a pressing need to search for novel Mtb drug targets in order to be able to combat these drug resistant strains. The enzyme 7,8-diaminopelargonic acid aminotransferase enzyme, or more commonly known as BioA, is one such ideal target, as it is known that humans do not possess this enzyme. BioA primarily plays a key role in Mtb’s lipid biosynthesis pathway; more specifically in the synthesis of the enzyme cofactor biotin. In this study, structure-based pharmacophore screening, docking, and ADMET evaluation of compounds obtained from the DrugBank chemical database were performed against the MtbBioA enzyme. Results of the screening, docking, ADMET, and TOPKAT calculations revealed that out of the 6,516 compounds in the library, only 7 compounds indicated more favorable binding energies as compared to the enzyme’s known inhibitor, amiclenomycin (ACM), as well as good solubility and toxicity properties. Moreover, out of these 7 compounds, Molecule 6 exhibited the best solubility and toxicity properties. In the future, these lead compounds may then be subjected to bioactivity assays in vitro or in vivo for further evaluation of its therapeutic efficacy.

Keywords: 7, 8-diaminopelargonic acid aminotransferase, BioA, pharmacophore, molecular docking, ADMET, TOPKAT

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Authors: Paula Buldres, Jorge Toledo

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Objectives: This study aimed to characterize potential probiotic strains isolated from canine breast milk for use in dogs with enteropathies. Methodology: Six canine breast milk strains, one canine colostrum strain, and one control porcine breast milk strain were characterized. According to its functional properties of resistance to acids, different concentrations of bile salts, and pancreatin, its presumptive properties of safety and inhibitory effect on pathogens, non-cytotoxic characteristics, and adhesion to the intestine. The immunomodulatory effect of formulations with better probiotic characterization in vitro and in vivo was also analyzed. Results: Two strains characterized as potential probiotics were obtained, which corresponded to the canine strains (TUCO-16 and TUCO-17), presenting resistance to acidic pH, bile salts, and pancreatin, as well as an inhibitory effect on pathogenic Escherichia coli, Salmonella sp., and Clostridium perfringens. Strains TUCO-16 and TUCO-17 induced a significant increase in the expression of TNF-α and IL-8 in canine macrophages, respectively. Expression analyses of pattern recognition receptors in DH82 cells suggest that TUCO-16 and TUCO-17 might increase the TLR2 expression marker, and porcine strain (TUCO-4) increases the NOD2 expression marker. Based on the count obtained and the encapsulation yield, the best formulations correspond to FOS-Inulin for the TUCO-17 and TUCO-4 strains; Maltodextrin-Inulin for TUCO-16. All the strains are non-cytotoxic. The strain that showed the highest adhesion to intestinal epithelial cells was TUCO-17 with the FOS-Inulin formulation. On the other hand, the probiotics decreased the expression of pro-inflammatory markers in vivo, both in the intestine and in the spleen of mice. Conclusion: The combination of these three strains under study (TUCO-16, TUCO-17, and TUCO-4) would cover the probiotic properties in formulation and immunomodulation of all the markers under study.

Keywords: probiotics, gastrointestinal infec, dog, probiotic formulation, immunomodulatory probiotics

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Authors: Pavana Basavakumar, Devadas Bhat

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Diabetes is a disease which often goes undiagnosed until its secondary effects are noticed. Early detection of the disease is necessary to avoid serious consequences which could lead to the death of the patient. Conventional invasive tests for screening of diabetes are mostly painful, time consuming and expensive. There’s also a risk of infection involved, therefore it is very essential to develop non-invasive methods to screen and estimate the level of blood glucose. Extensive research is going on with this perspective, involving various techniques that explore optical, electrical, chemical and thermal properties of the human body that directly or indirectly depend on the blood glucose concentration. Thus, non-invasive blood glucose monitoring has grown into a vast field of research. In this project, an attempt was made to device a prototype for screening of diabetes by measuring electrical impedance of the skin and building a model to predict a patient’s condition based on the measured impedance. The prototype developed, passes a negligible amount of constant current (0.5mA) across a subject’s index finger through tetra polar silver electrodes and measures output voltage across a wide range of frequencies (10 KHz – 4 MHz). The measured voltage is proportional to the impedance of the skin. The impedance was acquired in real-time for further analysis. Study was conducted on over 75 subjects with permission from the institutional ethics committee, along with impedance, subject’s blood glucose values were also noted, using conventional method. Nonlinear regression analysis was performed on the features extracted from the impedance data to obtain a model that predicts blood glucose values for a given set of features. When the predicted data was depicted on Clarke’s Error Grid, only 58% of the values predicted were clinically acceptable. Since the objective of the project was to screen diabetes and not actual estimation of blood glucose, the data was classified into three classes ‘NORMAL FASTING’,’NORMAL POSTPRANDIAL’ and ‘HIGH’ using linear Support Vector Machine (SVM). Classification accuracy obtained was 91.4%. The developed prototype was economical, fast and pain free. Thus, it can be used for mass screening of diabetes.

Keywords: Clarke’s error grid, electrical impedance of skin, linear SVM, nonlinear regression, non-invasive blood glucose monitoring, screening device for diabetes

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Authors: Anoosh Eghdami

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Background and aim: Conventional anticancer drugs display significant shortcomings which limit their use in cancer therapy. For this reason, important progress has been achieved in the field of nanotechnology to solve these problems and offer a promising and effective alternative for cancer treatment. Tumor markers may also be measured periodically during cancer therapy. Tumor markers may also be measured after treatment has ended to check for recurrence the return of cancer. The aim of this study was to evaluate the effect of nano drug delivery in induced breast cancer with DMBA by using CA15-3 tumor marker. Material and method: the rats were divided into five groups. The first group (control n=15) were fed only sesame oil as a gavage. In the second group n=15,10 mg DMBA was dissolved in 5ml of sesame oil and were fed as a gavage. In addition to DMBA treatment as the second group, in the 3,4and 5 groups after cancer creation, respectively affected by alpha interferon (α-IFN),alpha interferon conjugated with single walled carbon nano tube (α-IFN-SWNT) and encapsulated in poly lactic poly glycolic acid (α-IFN-PLGA). Tumor marker was measured in recent three groups. Results: The ANOVA test was used to determine the differences among the groups. Cancer inducing in rats (group 2) caused a significant increase in blood levels of CA15-3 (P<0.05). Administration of α-IFN, α-IFN –SWNT and α-IFN-PLGA in 3 groups of cancerous rats caused a significant decrease in blood levels of CA15-3 only the group that treated with α-IFN-PLGA (p<0.05). Conclusion: the results of this study indicate that nano drugs more effective than traditional drug in cancer treatment, although further work is needed to elucidate the safety and side effect of these compound in human.

Keywords: breast cancer, nano drug, tumor markers, CA15-3, α-IFN-PLGA, -IFN –SWNT

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Authors: Ratnawati Mohd Asraf, Hazlina Abdullah

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Numerous studies on boys’ performance relative to girls’ have been conducted around the globe. However, little has been done in relation to the literacy of primary school boys in the Malaysian context. This paper discusses the results of a study that sought to determine the literacy performance of Grades 1, 2, and 3 primary school students in the state of Selangor, Malaysia. Data on approximately 85,000 students from each grade level were obtained from the Ministry of Education Malaysia, which conducts national screening on literacy and numeracy, or LINUS, in all government primary schools. Teachers’ views were also sought through focus group interviews and journal entries. The results show that although there is an overall improvement in literacy performance in the Malay language among the students as they go into Grades 2 and 3, girls are found to outperform boys in every screening for all grade levels.

Keywords: boys’ underperformance, literacy, literacy performance, reading

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Authors: Ruolan Zhang, Ryo Imai, Naoko Senda, Tomoyuki Sakai

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Enzymes have attracted increasing attentions in industrial manufacturing for their applicability in catalyzing complex chemical reactions under mild conditions. Directed evolution has become a powerful approach to optimize enzymes and exploit their full potentials under the circumstance of insufficient structure-function knowledge. With the incorporation of cell-free synthetic biotechnology, rapid enzyme synthesis can be realized because no cloning procedure such as transfection is needed. Its open environment also enables direct enzyme measurement. These properties of cell-free biotechnology lead to excellent throughput of enzymes generation. However, the capabilities of current screening methods have limitations. Fluorescence-based assay needs applicable fluorescent label, and the reliability of acquired enzymatic activity is influenced by fluorescent label’s binding affinity and photostability. To acquire the natural activity of an enzyme, another method is to combine pre-screening step and high-performance liquid chromatography (HPLC) measurement. But its throughput is limited by necessary time investment. Hundreds of variants are selected from libraries, and their enzymatic activities are then identified one by one by HPLC. The turn-around-time is 30 minutes for one sample by HPLC, which limits the acquirable enzyme improvement within reasonable time. To achieve the real high-throughput enzyme screening, i.e., obtain reliable enzyme improvement within reasonable time, a widely applicable high-throughput measurement of enzymatic reactions is highly demanded. Here, a high-throughput screening method using broadband coherent anti-Stokes Raman spectroscopy (CARS) was proposed. CARS is one of coherent Raman spectroscopy, which can identify label-free chemical components specifically from their inherent molecular vibration. These characteristic vibrational signals are generated from different vibrational modes of chemical bonds. With the broadband CARS, chemicals in one sample can be identified from their signals in one broadband CARS spectrum. Moreover, it can magnify the signal levels to several orders of magnitude greater than spontaneous Raman systems, and therefore has the potential to evaluate chemical's concentration rapidly. As a demonstration of screening with CARS, alcohol dehydrogenase, which converts ethanol and nicotinamide adenine dinucleotide oxidized form (NAD+) to acetaldehyde and nicotinamide adenine dinucleotide reduced form (NADH), was used. The signal of NADH at 1660 cm⁻¹, which is generated from nicotinamide in NADH, was utilized to measure the concentration of it. The evaluation time for CARS signal of NADH was determined to be as short as 0.33 seconds while having a system sensitivity of 2.5 mM. The time course of alcohol dehydrogenase reaction was successfully measured from increasing signal intensity of NADH. This measurement result of CARS was consistent with the result of a conventional method, UV-Vis. CARS is expected to have application in high-throughput enzyme screening and realize more reliable enzyme improvement within reasonable time.

Keywords: Coherent Anti-Stokes Raman Spectroscopy, CARS, directed evolution, enzyme screening, Raman spectroscopy

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Authors: Abdelrahman A. Ramzy, Bassel S. Abdallah, Mohamed E. Bahgat, Sarah M. Abdelkader, Sherif H. ElGohary

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Attention-Deficit/Hyperactivity Disorder (ADHD), epilepsy, and autism affect millions of children worldwide, many of which are undiagnosed despite the fact that all of these disorders are detectable in early childhood. Late diagnosis can cause severe problems due to the late treatment and to the misconceptions and lack of awareness as a whole towards these disorders. Moreover, electroencephalography (EEG) has played a vital role in the assessment of neural function in children. Therefore, quantitative EEG measurement will be utilized as a tool for use in the evaluation of patients who may have ADHD, epilepsy, and autism. We propose a screening tool that uses EEG signals and machine learning algorithms to detect these disorders at an early age in an automated manner. The proposed classifiers used with epilepsy as a step taken for the work done so far, provided an accuracy of approximately 97% using SVM, Naïve Bayes and Decision tree, while 98% using KNN, which gives hope for the work yet to be conducted.

Keywords: ADHD, autism, epilepsy, EEG, SVM

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