Expression of TGF-β1 in the blood during fracture repair in an estrogen-deficient rat model

OBJECTIVES: Previous studies have reported that osteoporosis due to estrogen deficiency influences fracture healing. Transforming growth factor (TGF-b) has been found to be involved in fracture healing via the regulation of the differentiation and activation of osteoblasts and osteoclasts. The current study aimed to determine the effects of estrogen on the expression of TGF-&#946;1 during fracture healing in ovariectomized rats. METHODS: Thirty female Sprague-Dawley rats weighing 200-250 g were assigned to: (i) a sham-operated group that was given a normal saline; (ii) an ovariectomized control group that was given a normal saline; or (iii) an ovariectomized + estrogen (100 mg/kg/day) group that was treated with conjugated equine estrogen. The right femur of all rats was fractured, and a Kirschner wire was inserted six weeks post-ovariectomy. Treatment with estrogen was given for another six weeks post-fracture. At the end of the study, blood samples were taken, and the right femur was harvested and subjected to biomechanical strength testing. RESULTS: The percentage change in the plasma TGF-&#946;1 level before treatment was significantly lower in the ovariectomized control and estrogen groups when compared with the sham group (p<0.001). After six weeks of treatment, the percentage change in the plasma TGF-&#946;1 level in the estrogen group was significantly higher compared with the level in the ovariectomized control group (p = 0.001). The mean ultimate force was significantly increased in the ovariectomized rats treated with estrogen when compared with the ovariectomized control group (p = 0.02). CONCLUSION: These data suggest that treatment with conjugated equine estrogen enhanced the strength of the healed bone in estrogen-deficient rats by most likely inducing the expression of TGF-&#946;1

Evaluation of in vitro wound healing efficacy of breadfruit derived starch hydrolysate

To study the wound healing efficacy of breadfruit starch hydrolysate, an in vitro wound scratch assay was conducted, in which the migration rate of wounded NIH 3T3 fibroblasts was determined. Wounds treated with lower dextrose equivalent (DE), (DE 10-14) starch hydrolysate were found capable to improve the wound healing of NIH 3T3 fibroblast cell with the percentage of wound closure improvement of 77%, respectively when compared with higher DE range (DE 15-19 and DE 20-24). The findings obtained in the BrdU uptake and MTT viability assays confirmed the wound healing properties of breadfruit starch hydrolysate as the starch hydrolysate-treated wounded NIH 3T3 fibroblasts were able to proliferate well and no cytotoxicity was observed. Together, these findings indicated that the newly developed breadfruit starch hydrolysate performed better than commercial (COM) starch hydrolysate of the same DE ranges. In conclusion, breadfruit starch hydrolysate had better functional properties than did starch hydrolysates derived from other sources and that they could play a beneficial role in wound healing applications

Oncolytic effects of the recombinant newcastle disease virus, rAF-IL12, against colon cancer cells in vitro and in tumor-challenged NCr-foxn1nu nude mice

Colon cancer remains one of the main cancers causing death in men and women worldwide as certain colon cancer subtypes are resistant to conventional treatments and the development of new cancer therapies remains elusive. Alternative modalities such as the use of viral-based therapeutic cancer vaccine is still limited, with only the herpes simplex virus (HSV) expressing granulocyte-macrophage colony- stimulating factor (GM-CSF) or talimogene laherparepvec (T-Vec) being approved in the USA and Europe so far. Therefore, it is imperative to continue the search for a new treatment modality. This current study evaluates a combinatorial therapy between the oncolytic Newcastle disease virus (NDV) and interleukin-12 (IL-12) cytokine as a potential therapeutic vaccine to the current anti-cancer drugs. Several in vitro analyses such as MTT assay, Annexin V/FITC flow cytometry, and cell cycle assay were performed to evaluate the cytotoxicity effect of recombinant NDV, rAF-IL12. Meanwhile, serum cytokine, serum biochemical, histopathology of organs and TUNEL assay were carried out to assess the anti-tumoral effects of rAF-IL12 in HT29 tumor-challenged nude mice. The apoptosis mechanism underlying the effect of rAF-IL12 treatment was also investigated using NanoString Gene expression analysis. The recombinant NDV, rAF-IL12 replicated in HT29 colon cancer cells as did its parental virus, AF2240-i. The rAF-IL12 treatment had slightly better cytotoxicity effects towards HT29 cancer cells when compared to the AF2240-i as revealed by the MTT, Annexin V FITC and cell cycle assay. Meanwhile, the 28-day treatment with rAF-IL12 had significantly (p < 0.05) perturbed the growth and progression of HT29 tumor in NCr-Foxn1nu nude mice when compared to the untreated and parental wild-type NDV strain AF2240-i. The rAF-IL12 also modulated the immune system in nude mice by significantly (p < 0.05) increased the level of IL-2, IL-12, and IFN-γ cytokines. Treatment with rAF-IL12 had also significantly (p < 0.05) increased the expression level of apoptosis-related genes such as Fas, caspase-8, BID, BAX, Smad3 and granzyme B in vitro and in vivo. Besides, rAF-IL12 intra-tumoral delivery was considered safe and was not hazardous to the host as evidenced in pathophysiology of the normal tissues and organs of the mice as well as from the serum biochemistry profile of liver and kidney. Therefore, this study proves that rAF-IL12 had better cytotoxicity effects than its parental AF2240-i and could potentially be an ideal treatment for colon cancer in the near future

Safety and efficacy of recombinant newcastle disease virus expressing human interleukin-12 as a potential vaccine in breast cancer

In this developing era, breast cancer still remains a life-threatening disease globally. However, oncolytic virotherapy has taken over interest as a promising non-conventional alternative to treating breast cancers. Newcastle disease virus (NDV), an avian paramyxovirus has been demonstrated with significant oncolytic activity against cancer based on numerous preclinical studies. Today, genetically modified viruses coding for immunomodulatory agents, such as cytokines or chemokines, have come into focus. Such engineered viruses are able to promote efficient immune responses against tumour cells. The overall of this project aims to study the effects of a recombinant NDV expressing human interleukin 12 (rAF-IL12) in the apoptotic and metastatic process in MCF7 and MDA-MB231 human breast cancer cell lines. The parental NDV AF2240 was used as a positive control in this study. Notably, rAF-IL12 was able to maintain its stability when passaged in specific pathogen free (SPF) eggs up to ten passages. Furthermore it is considered safe as it selectively induced cytotoxic effects in chicken and breast cancer cell lines while sparing non-cancerous breast cell line as demonstrated through the MTT assay. The stability of each passaged rAF-IL12 was verified via haemagglutination assay (HA), mean death time (MDT) and intracerebral pathogenicity index (ICPI), while the IL12 was quantified through Enzyme-Linked Immunosorbent Assay (ELISA). Comparable to AF2240, rAF-IL12 was also able to induce apoptosis significantly based on several apoptotic assays. Both rAF-IL12 and AF2240 managed to increase the percentage of G2/M and S phase in the cell cycle analysis while inducing the percentage of apoptosis. Although both AF2240 and rAF-IL12 demonstrated comparable in vitro apoptosis results, rAF-IL12 possessed significant (p<0.05) antimetastatic activity in comparison to AF2240 based on metastasis related assays including the in vitro scratch assay, migration/ invasion assay, human umbilical vein endothelial cell (HUVEC) tube formation and rat aortic ring assay. Additionally, to further evaluate the anti-tumour and anti-metastatic mechanism of rAF-IL12, in vivo studies were conducted using 4T1-challenged BALB/c mice as a model of this study. The rAF-IL12 was proven to function as an improved tumour vaccine as it significantly (p<0.05) reduced the size of tumour in comparison to the parental AF2240 virus. Apoptotic results showed that the number of cancer cells in the tumour significantly (p<0.05) reduced after 28 days of intra-tumoural treatment with rAF-IL12 (27 HAU). In addition, rAF-IL12 was able to inhibit the migration of cancer cells to other vital organs as opposed to the untreated group. To further elucidate the apoptotic and anti-metastatic mechanism of rAF-IL12 at molecular level, NanoString nCounter was conducted. Even though both AF2240 and rAF-IL12 exhibited similar mechanism of action, rAF-IL12 was more potent than AF2240 in terms of apoptosis and anti- metastasis activity. In conclusion, both rAF-IL12 and AF2240 were able to inhibit the proliferation of breast cancer cells and induce apoptosis in the in vitro studies; however, rAF-IL12 was able to demonstrate significant (p<0.05) improved functionality in comparison to AF2240 alone based on the in vivo studies. This proved that interleukin 12 was able to increase the immune response against tumour cells by inducing cell death, anti-angiogenesis and anti-metastasis effects, further improving the function of AF2240 as a potential vaccine in breast cancer

Percutaneous Catheter-Based Rheolytic Thrombectomy for Massive Pulmonary Embolism: A Case Report

Pulmonary thromboembolism is a life-threatening cardiovascular condition. The mortality rate is high in its current management. Besides supportive treatments, systemic thrombolysis and surgical thrombectomy play important roles in the comprehensive management of pulmonary embolism (PE). The percutaneous catheter-based rheolytic thrombectomy is a promising alternative for management of massive pulmonary emboli, particularly, when patients have contraindication with systemic thrombolysis or are not suitable for surgery. We present the case of a 36-year-old Somalian man who came to our center for a total knee replacement (TKR). Three days after TKR, he developed sudden shortness of breath and decreased oxygen saturation. Computed tomography of pulmonary arteriogram showed extensive thrombi within the main pulmonary trunk, right and left pulmonary arteries, bilateral ascending and bilateral descending pulmonary arteries in keeping with massive PE. Because the patient was contraindicated for systemic thrombolysis, percutaneous, catheter-based rheolytic thrombectomy was chosen as the alternative treatment. His clinical symptoms improved immediately post-treatment. In conclusion, catheter-based rheolytic thrombectomy can serve as an alternative treatment for massive PE with a good clinical outcome

An Evaluation of the Quality of Plain Radiograph Interpretations by Radiology Trainees: A Single Institution Experience

Introduction: The primary communication between the radiologist and referrer is through the radiological report. However, there are incidents of misinterpretation during radiologist training. Therefore, the present study aimed to evaluate the accuracy level and incidence of interpretation errors for plain radiographs among radiology trainees at our institution. Materials and Methods: The present study retrospectively reviewed 508 reported plain radiographs for one year, and two radiologists subsequently evaluated these plain radiographs. The initial diagnosis by the trainee was compared with the radiologists&rsquo; evaluation, and the results were categorized as either &lsquo;accurate&rsquo;, &lsquo;minor discrepancy&rsquo;, or &lsquo;major discrepancy&rsquo;. The data were analyzed concerning the overall performance, year of trainee, anatomic area, patient age group, and radiograph type. A chi-square test was performed, with p &lt; 0.05 indicating statistical significance. Results: The overall accuracy rate was 69%, with minor and major discrepancy rates of 21% and 10%, respectively. There was an insignificant increase in overall accuracy with increased years of training, despite a reduction to 58% accuracy among Year 3 trainees. The accuracy level increased between Year 1, Year 2 and Year 4 by 70%, 71% and 75%, respectively (p &gt; 0.05). The accuracy rates for both the adult and pediatric age groups were not statistically significant. The mobile radiographs showed lower accuracy rate of reporting than the plain radiographs. Conclusion: The radiological trainee interpretations for plain radiographs had an average rating with low discrepancy rates. The Year 3 trainees had the lowest accuracy compared to the other trainee groups. However, the present study suggests the need for further research to determine if the current outcomes are outliers or are indicative of a real phenomenon

Inter-observer and intra-observer variability in the assessment of the paranasal sinuses radiographs

The aims of this study are to determine the intra-observer and inter-observer agreement between the two radiologists when reporting the Waters view and 3 views series of the paranasal sinus radiographs. To also determine whether a single Waters view is adequate for diagnostic purpose of sinusitis. 90 sets of paranasal sinuses radiographs were reported by two radiologists independently. The results revealed that the intra-observer and inter-observer agreement varied from almost perfect to good for maxillary and frontal sinuses, but poor for sphenoid and ethmoidal sinuses. Therefore, a single waters view is an acceptable strategy for diagnosing sinusitis for the maxillary and/or frontal sinuses but not for the ethmoid and sphenoid sinuses

Efficacy Study of Broken Rice Maltodextrin in In Vitro Wound Healing Assay

Maltodextrins that contain both simple sugars and polymers of saccharides have been widely used as ingredients in food products and pharmaceutical delivery systems. To date, no much work has been reported on the applications of maltodextrin from broken rice (RB) sources. Therefore, the objective of this work was to investigate the in vitro wound healing efficacy of RB maltodextrin at different conditions. Wounds treated with lower dextrose equivalent (DE) range (DE 10–14) of maltodextrins at a concentration of 10% obtained from RB were found to be able to heal the wounds significantly faster (p<0.01) than maltodextrin with higher DE ranges (DE 15–19 and DE 20–24) and concentrations of 5% and 20%. The findings from both BrdU and MTT assay further confirmed its wound healing properties as the NIH 3T3 fibroblast wounded cells were able to proliferate without causing cytotoxic effect when wounded cell was treated with maltodextrin. All these findings indicated that the RB maltodextrin could perform better than the commercial maltodextrin at the same DE range. This study showed that RB maltodextrins had better functionality properties than other maltodextrin sources and played a beneficial role in wound healing application