Ahmad Faried

Inhibition of autophagy augments 5-fluorouracil chemotherapy in human colon cancer in vitro and in vivo model.

Although 5-fluorouracil (5-FU)-based adjuvant chemotherapy is widely used in the treatment of colorectal cancer, novel therapeutic strategies need to be explored. It has been reported that autophagy is extensively implicated in cancer. However, the function of autophagy is not fully understood. In the present study, apoptosis induced by 5-FU in 3 human colon cancer cell lines (HCT116, DLD-1, and DLD-1/5-FU (a specific 5-FU-resistant sub-line)) was measured using MTT assay, DNA fragmentation assay, Hoechst 33342 staining, and caspase-3 immunoblotting. The autophagy activation induced by 5-FU treatment was revealed by microtubule-associated protein 1 light chain 3 (LC3) immunofluorescence and immunoblotting and p62 immunoblotting. Inhibition of autophagy by 3-methyladenine (3-MA) or small interference RNA targeting Atg7 (Atg7 siRNA) significantly augmented 5-FU-induced apoptosis. This synergistic effect of 5-FU and 3-MA was further confirmed in the DLD-1 xenograft tumour model. Tumour growth was suppressed more significantly with combination treatment than 5-FU treatment alone. In conclusion, autophagy was activated as a protective mechanism against 5-FU-induced apoptosis and its inhibition could be a promising strategy for adjuvant chemotherapy in colon cancer.

Selenium enrichment of broccoli sprout extract increases chemosensitivity and apoptosis of LNCaP prostate cancer cells

BackgroundBroccoli is a Brassica vegetable that is believed to possess chemopreventive properties. Selenium also shows promise as an anticancer agent. Thus, selenium enrichment of broccoli has the potential to enhance the anticancer properties of broccoli sprouts.MethodSelenium-enriched broccoli sprouts were prepared using a sodium selenite solution. Their anticancer properties were evaluated in human prostate cancer cell lines and compared with those of a control broccoli sprout extract.ResultsSelenium-enriched broccoli sprouts were superior to normal broccoli sprouts in inhibiting cell proliferation, decreasing prostate-specific antigen secretion, and inducing apoptosis of prostate cancer cells. Furthermore, selenium-enriched broccoli sprouts but, not normal broccoli sprouts, induced a downregulation of the survival Akt/mTOR pathway.ConclusionOur results suggest that selenium-enriched broccoli sprouts could potentially be used as an alternative selenium source for prostate cancer prevention and therapy.

Experimental investigation of encephalomyosynangiosis using gyrencephalic brain of the miniature pig: histopathological evaluation of dynamic reconstruction of vessels for functional anastomosis. Laboratory investigation.

OBJECT Encephalomyosynangiosis (EMS) is a surgical treatment for moyamoya disease that is widely used to provide increased intracranial blood flow via revascularization by arterial anastomosis from the external carotid artery. However, the angiogenic mechanism responsible for the revascularization induced by EMS has not been systematically evaluated. In this study the authors investigated the chronological angiogenic changes associated with EMS to clarify the favorable factors and identify revascularization mechanisms by using an experimental internal carotid artery occlusion (ICAO) model in the miniature pig. METHODS Fourteen miniature pigs were used, 11 of which underwent ICAO before transcranial surgery for EMS was performed. Animals were allowed to recover for 1 week (4 pigs) or 4 weeks (7 pigs) after EMS. Control group animals were treated in the same way, but without occlusion (3 pigs). Magnetic resonance imaging, angiography, and histological investigation were performed. RESULTS One week after EMS, on histological examination of both the ICAO and control groups it was found that the transplanted temporal muscle had adhered to the arachnoid via a granulation zone, which was enriched with immune cells such as macrophages associated with the angiogenic process. Four weeks after EMS, angiography and histological examination of the ICAO group showed patent anastomoses between the external carotid artery and the cortical arteries without any detectable boundary between the temporal muscle and the cerebral cortex. In contrast, histological examination of the control group found scar tissue between the cerebral cortex and temporal muscle. CONCLUSIONS The initial step for formation of anastomoses resembles the process of wound healing associated with repair processes such as active proliferation of macrophages and angiogenesis within the new connective tissue. Functional revascularization requires a suitable environment (such as tissue containing vascular beds) and stimulus (such as ischemia) to induce vascular expansion.

Chemically synthesized sugar‐cholestanols possess a preferential anticancer activity involving promising therapeutic potential against human esophageal cancer

The understanding of the cell signaling pathways and the molecular events leading to cell death of cancer cells will provide in‐depth perspective into the identification and development of potent anticancer agents. A balance between cell proliferation and cell death has been raised as a rational target for the management of malignant tumors. In the present study, the authors demonstrated that chemically synthesized sugar‐cholestanols consisting of GlcNAcβ‐, Galβ‐ and GlcNAcβ1,3Galβ‐cholestanols exerted a strong inhibiting activity against cell proliferation of esophageal cancer cells, but cholestanol itself did not show such an activity against the same cancer cells at all. In addition to their predominant role as an antiproliferation agent, evidence based on the molecular analyses suggested that sugar‐cholestanols played a regulatory role in multiple signal transduction pathways inducing apoptosis through both the death signal‐extrinsic and the mitochondria‐intrinsic pathways. Sugar‐cholestanols seemed to be more susceptible to esophageal cancer cells than to non‐cancerous esophageal cells at the very early event of their exposure and, further, to suppress specifically the expression of vascular endothelial growth factor. Taken together, these novel functions of sugar‐cholestanols indicate that they could have promising therapeutic potential against human esophageal cancer. (Cancer Sci 2007; 98: 1358–1367)

A comparison of cryopreservation methods: Slow-cooling vs. rapid-cooling based on cell viability, oxidative stress, apoptosis, and CD34+ enumeration of human umbilical cord blood mononucleated cells

BackgroundThe finding of human umbilical cord blood as one of the most likely sources of hematopoietic stem cells offers a less invasive alternative for the need of hematopoietic stem cell transplantation. Due to the once-in-a-life time chance of collecting it, an optimum cryopreservation method that can preserve the life and function of the cells contained is critically needed.MethodsUntil now, slow-cooling has been the routine method of cryopreservation; however, rapid-cooling offers a simple, efficient, and harmless method for preserving the life and function of the desired cells. Therefore, this study was conducted to compare the effectiveness of slow- and rapid-cooling to preserve umbilical cord blood of mononucleated cells suspected of containing hematopoietic stem cells. The parameters used in this study were differences in cell viability, malondialdehyde content, and apoptosis level. The identification of hematopoietic stem cells themselves was carried out by enumerating CD34+ in a flow cytometer.ResultsOur results showed that mononucleated cell viability after rapid-cooling (91.9%) was significantly higher than that after slow-cooling (75.5%), with a p value = 0.003. Interestingly, the malondialdehyde level in the mononucleated cell population after rapid-cooling (56.45 μM) was also significantly higher than that after slow-cooling (33.25 μM), with a p value < 0.001. The apoptosis level in rapid-cooling population (5.18%) was not significantly different from that of the mononucleated cell population that underwent slow-cooling (3.81%), with a p value = 0.138. However, CD34+ enumeration was much higher in the population that underwent slow-cooling (23.32 cell/μl) than in the one that underwent rapid-cooling (2.47 cell/μl), with a p value = 0.001.ConclusionsRapid-cooling is a potential cryopreservation method to be used to preserve the umbilical cord blood of mononucleated cells, although further optimization of the number of CD34+ cells after rapid-cooling is critically needed.

Penetrating skull fracture by a wooden object: Management dilemmas and literature review

Most penetrating skull injuries are caused by gun shot wounds or missiles. The compound depressed skull fracture represents an acute neurosurgical emergency. Management and diagnosis of such cases have been described, but its occurence following a fall onto a piece of wood is quite unusual. A 75-year-old female fell onto a piece of wood that penetrated her skull on the left frontal region and was treated in our department. The patient had no neurological deficits during presentation. She was managed surgically and removal of the wooden object was performed to prevent early or late infection complications. Wooden foreign bodies often pose a different set of challenges as far as penetrating injuries to the brain are concerned. Radiological difficulties and increased rates of infection due to its porous nature make these types of injuries particularly interesting. Their early diagnosis and appropriate treatment can minimize the risk of complications.

Expression of Carbohydrate Antigens in Human Esophageal Squamous Cell Carcinoma: Prognostic Application and Its Diagnostic Implications

BackgroundTumor markers whose antigenic determinants have been demonstrated to consist of carbohydrates are probably one of the most extensive tools that have been used in routine cancer diagnosis. In this study, the relevance of carbohydrate antigen expression profile was examined in esophageal squamous cell carcinoma together with prognosis in 130 patients.MethodsThe expression of carbohydrate antigens was estimated immunohistochemically by anti–sialyl Lewis a (sialyl Lea) and anti–sialyl Lewis x (sialyl Lex) monoclonal antibodies, and correlation between their staining and clinicopathological status was examined. In addition, the correlation of both carbohydrate antigens expression was evaluated with microvessel density (MVD).ResultsExpressions of sialyl Lewis antigens and MVD were associated with several clinicopathological features that reflect the tumor aggressiveness in esophageal cancer. The 5-year survival rate of patients was found to be associated with expression of sialyl Lea and sialyl Lex antigens and with MVD; thus, all of them were revealed to be independent prognostic factors.ConclusionsCombination of these factors offered a better prediction of prognosis of esophageal squamous cell carcinoma. Further, carbohydrate antigens represent a promising target for therapeutic approaches against the disease.

Trumpet Laminectomy Microdecompression for Lumbal Canal Stenosis

Microsurgery techniques are useful innovations towards minimizing the insult of canal stenosis. Here, we describe the trumpet laminectomy microdecompression (TLM) technique, advantages and disadvantages. Sixty-two TLM patients with lumbar disc herniation, facet hypertrophy or yellow ligament or intracanal granulation tissue. The symptoms are low back pain, dysesthesia and severe pain on both legs. Spine levels operated Th11-S1; the patients who had trumpet-type fenestration, 62.9% had hypertrophy of the facet joint, 11.3% had intracanal granulation tissue, 79.1% had hypertrophy of the yellow ligament and 64.5% had disc herniation. The average of procedure duration was 68.9 min and intraoperative blood loss was 47.4 mL. Intraoperative complications were found in 3.2% of patients, with dural damage but without cerebrospinal fluid leakage. The TLM can be performed for all ages and all levels of spinal canal stenosis, without the complication of spondilolistesis. The TLM has a shorter duration, with minimal intraoperative blood loss.

Inhibition of activated NR2B gene- and caspase-3 protein-expression by glutathione following traumatic brain injury in a rat model

Background. Traumatic brain injury (TBI) remains a major cause of death and disability. Oxidative stress is an important element of the injury cascade following TBI. Progressive compromise of antioxidant defenses and free radical-mediated lipid peroxidation are one of the major mechanisms of secondary TBI. NR2B is a glutamate receptor and its activation is caused by TBI increasing a brain cell death, along with caspase-3 as a hall mark of apoptosis. Glutathione is a potent free radical scavenger that might prevent secondary TBI damage and inhibited apoptosis. Materials and Methods. In the present study, it aims to demonstrate the effect of glutathione on inhibition of brain oxidative damage in a TBI rat model. Results. In this study, the expressions of mRNA NR2B in placebo group and groups with glutathione administration at 0, 3, and 6 hours after TBI were 328.14, 229.90, 178.50, and 136.14, respectively (P<0.001). The highest caspase-3 expression was shown in placebo group with 66.7% showing strong positive results (>80%); as expected, glutathione administered in 0, 3, and 6 hours groups had lower strong positive results of 50%, 16.7%, and 16.7%, respectively, (P=0.025). Conclusion. In conclusion, this study showed that glutathione administration in a TBI rat model decreased NR2B gene- and caspase-3 protein-expression that lead to the inhibition of brain cell death. Our results suggest that glutathione, as a potent free radical scavenger, has a brain cell protective effect against oxidative damage and cell death induced by TBI in rat model.

Characteristics of Moderate and Severe Traumatic Brain Injury of Motorcycle Crashes in Bandung, Indonesia

BACKGROUND Traumatic brain injury (TBI) constitutes a critical public health and socioeconomic problem. As one of the leading causes of mortality and disability from road traffic crashes, the incidence of TBIs is increasing with increasing motor vehicle usage. Understanding the prevalence and describing the characteristics of TBI are crucial for successful implementation of prevention and treatment efforts to reduce the mortality and morbidity caused by TBIs. METHODS We studied cases of moderate and severe TBI resulting from motorcycle crashes from January 1, 2013 to June 30, 2014. Variables studied included sex, age, time interval (from crash to arrival at the emergency department), alcohol consumption, helmet use, severity of TBI, choice of treatment, and the outcome. RESULTS A total of 2108 head injury cases were seen at the emergency department during this period, 1324 (62.8%) of which resulted from motorcycle crashes. Of those cases, 30.7% (407 cases) were categorized as moderate or severe TBI with 29.2% mortality. Most of the patients were male (80.8%), <60 years old (96.1%), and did not wear a helmet (71.2%). More than half of the cases (56.7%) arrived at the emergency department within 6 hours, 14.0% of the cases were under alcohol intoxication, and 37.8% of the cases were operated on. CONCLUSIONS This preliminary analysis highlights the need to address road safety, especially with respect to helmet use and drink driving, to reduce the burden of TBIs in Bandung.