ETM study of electroporation influence on cell morphology in human malignant melanoma and human primary gingival fibroblast cells

Objective

To estimate electroporation (EP) influence on malignant and normal cells.

Methods

Two cell lines including human malignant melanoma (Me-45) and normal human gingival fibroblast (HGFs) were used. EP parameters were the following: 250, 1 000, 1 750, 2 500 V/cm; 50 µs by 5 impulses for every case. The viability of cells after EP was estimated by MTT assay. The ultrastructural analysis was observed by transmission electron microscope (Zeiss EM 900).

Results

In the current study we observed the intracellular effect following EP on Me-45 and HGF cells. At the conditions applied, we did not observe any significant damage of mitochondrial activity in both cell lines treated by EP. Conversely, we showed that EP in some conditions can stimulate cells to proliferation. Some changes induced by EP were only visible in electron microscopy. In fibroblast cells we observed significant changes in lower parameters of EP (250 and 1 000 V/cm). After applying higher electric field intensities (2 500 V/cm) we detected many vacuoles, myelin-like bodies and swallowed endoplasmic reticulum. In melanoma cells such strong pathological modifications after EP were not observed, in comparison with control cells. The ultrastructure of both treated cell lines was changed according to the applied parameters of EP.

Conclusions

We can claim that EP conditions are cell line dependent. In terms of the intracellular morphology, human fibroblasts are more sensitive to electric field as compared with melanoma cells. Optimal conditions should be determined for each cell line. Summarizing our study, we can conclude that EP is not an invasive method for human normal and malignant cells. This technique can be safely applied in chemotherapy for delivering drugs into tumor cells.     

An improved model for predicting postoperative nausea and vomiting in ambulatory surgery patients using physician-modifiable risk factors

Objective

Postoperative nausea and vomiting (PONV) is a frequent complication in patients undergoing ambulatory surgery, with an incidence of 20%–65%. A predictive model can be utilized for decision support and feedback for practitioner practice improvement. The goal of this study was to develop a better model to predict the patient''s risk for PONV by incorporating both non-modifiable patient characteristics and modifiable practitioner-specific anesthetic practices.

Materials and methods

Data on 2505 ambulatory surgery cases were prospectively collected at an academic center. Sixteen patient-related, surgical, and anesthetic predictors were used to develop a logistic regression model. The experimental model (EM) was compared against the original Apfel model (OAM), refitted Apfel model (RAM), simplified Apfel risk score (SARS), and refitted Sinclair model (RSM) by examining the discriminating power calculated using area under the curve (AUC) and by examining calibration curves.

Results

The EM contained 11 input variables. The AUC was 0.738 for the EM, 0.620 for the OAM, 0.629 for the RAM, 0.626 for the SARS, and 0.711 for the RSM. Pair-wise discrimination comparison of models showed statistically significant differences (p<0.05) in AUC between the EM and all other models, OAM and RSM, RAM and RSM, and SARS and RSM.

Discussion

All models except the OAM appeared to have good calibration for our institution''s ambulatory surgery data. Ours is the first model to break down risk by anesthetic technique and incorporate risk reduction due to PONV prophylaxis.

Conclusion

The EM showed statistically significant improved discrimination over existing models and good calibration. However, the EM should be validated at another institution.     

Revisiting the Boyne procedure

Background

The technique involving the free grafting of the mandibular condyle with a vertical ramus osteotomy was described by Philip Boyne in 1987 along with a series of 15 cases where a patient presented with a markedly displaced fracture dislocation of the mandibular condyle.

Method

Twenty-one cases (25 condylar fractures) were treated with a modified Boyne technique.

Results

Results showed that we had a complication rate of 12% (8% resorption, 4% fracture of the plate).

Conclusion

The Boyne procedure should be thought of as a mainstream treatment method for the displaced fractured mandibular condyle.Key Words: Boyne procedure     

Isolation and efficacy of entomopathogenic fungus (Metarhizium anisopliae) for the control of Aedes albopictus Skuse larvae: suspected dengue vector in Pakistan

Objective

To isolate the entomopathogenic fungus Metarhizium anisopliae (M. anisopliae) in the local environment, and evaluate its efficacy against the suspected dengue vector Aedes albopictus in Pakistan.

Methods

According to the standard procedure, M. anisopliae was isolated from the dead mosquitoes which were collected from the field or dead after the collection. Bioassay was performed to determine its efficacy.

Results

The results indicated that M. anisopliae had larvicidal effect with LC₅₀ value 1.09×10⁵ and LC₉₀ value 1.90×10¹³ while it took 45.41 h to kill 50% of tested population.

Conclusions

Taking long time to kill 50% population when compare with the synthetic insecticides, is the only drawback for the use of entomopathogenic fungus but these bio-pesticides are safe for the use.     

Can poison control data be used for pharmaceutical poisoning surveillance?

Objective

To determine the association between the frequencies of pharmaceutical exposures reported to a poison control center (PCC) and those seen in the emergency department (ED).

Design

A statewide population-based retrospective comparison of frequencies of ED pharmaceutical poisonings with frequencies of pharmaceutical exposures reported to a regional PCC. ED poisonings, identified by International Classification of Diseases, Version 9 (ICD-9) codes, were grouped into substance categories. Using a reproducible algorithm facilitated by probabilistic linkage, codes from the PCC classification system were mapped into the same categories. A readily identifiable subset of PCC calls was selected for comparison.

Measurements

Correlations between frequencies of quarterly exposures by substance categories were calculated using Pearson correlation coefficients and partial correlation coefficients with adjustment for seasonality.

Results

PCC reported exposures correlated with ED poisonings in nine of 10 categories. Partial correlation coefficients (r_p) indicated strong associations (r_p>0.8) for three substance categories that underwent large changes in their incidences (opiates, benzodiazepines, and muscle relaxants). Six substance categories were moderately correlated (r_p>0.6). One category, salicylates, showed no association.

Limitations

Imperfect overlap between ICD-9 and PCC codes may have led to miscategorization. Substances without changes in exposure frequency have inadequate variability to detect association using this method.

Conclusion

PCC data are able to effectively identify trends in poisonings seen in EDs and may be useful as part of a pharmaceutical poisoning surveillance system. The authors developed an algorithm-driven technique for mapping American Association of Poison Control Centers codes to ICD-9 codes and identified a useful subset of poison control exposures for analysis.     

Mortality of therapeutic fish Garra rufa caused by Aeromonas sobria

Objective

To investigate a case of mass mortality of Garra rufa (G. rufa) from a fish hatchery farm in Slovakia.

Methods

Causative bacterial agent was swabbing out of affected fish skin area and subsequently identified using commercial test system. Antibiotic susceptibility was determined by the disk diffusion method.

Results

Infected G. rufa was characterized by abnormal swimming behaviour, bleeding of skin lesions and local haemorrhages. Despite of using recommended aquatic antibiotic treatment no improvement was achieved and Aeromonas sobria (A. sobria) was identified as a causative agent of fish mortality. Due to massive fish mortality, antibiotic susceptibility of pure isolated culture of A. sobria was evaluated employing eight antibiotics against human infections. A. sobria was resistant only against one antibiotic, namely ampicilin.

Conclusions

These results indicate that A. sobria can act as a primary pathogen of G. rufa and may be a potential risk factor for immunodeficient or immunoincompetent patients during the ichthyotherapy.     

Evaluation of Vitreous Substitutes in Managing Complicated Vitreo-retinal Surgeries

Background

Vitreous substitutes presently in use for intraoperative tamponade are perfluorocarbon liquids (PFCL) and for post operative tamponade are silicon oil (SO), sulphur hexafluoride (SF₆) and perfluorocarbon gas (PFC). Several factors are important for a thorough understanding of each of these vitreous substitutes. The absorption or necessity for removal, indications for use (including clinical studies and special surgical methods), additives and complications of use have to be considered.

Methods

Three port standard pars plana vitrectomy was performed in 60 consecutive cases requiring intra-vitreal intervention. PFCL was used intra-operatively in 30 cases (PFCL group and X group). The eyes were implanted with SO (silicon oil subgroup), PFC (C₃F₈ sub-group) and SF₆ (SF₆ sub-group) in twenty cases each. Apart from best corrected visual acuity (BCVA), the retinal status and the longevity of gas when used, changes and reaction in both the anterior and posterior segments were noted.

Result

The difference of postoperative BCVA in the PFCL and non-PFCL groups was statistically significant with p < 0.001. In both the PFCL and non-PFCL groups the difference between preoperative and postoperative BCVA was statistically significant with p < 0.0001. All three vitreous substitutes studied are effective given the case where they had been utilised.

Conclusion

PFCL is invaluable in the management of complicated retinal detachment (RD). Where a vitreous microsurgery is indicated, the visual outcome is good. SF₆ is useful for short-term tamponade. Silicon oil and C₃F₈ are useful for longer tamponade.Key Words: Tamponade, Perfluorocarbon liquids, Silicon oil, Sulphur hexafluoride, Perfluorocarbon gas, Retinal detachment     

A phase II study on safety and efficacy of high-dose N-acetylcysteine in patients with cystic fibrosis

Objective

We conducted a single-centre, randomised, double-blinded, placebo-controlled phase II clinical study to test safety and efficacy of a 12-week therapy with low-dose (700 mg/daily) or high-dose (2800 mg/daily) of NAC.

Methods

Twenty-one patients (ΔF508 homo/heterozygous, FEV₁ > 40% pred.) were included in the study. After a 3-weeks placebo run-in phase, 11 patients received low-dose NAC, and 10 patients received high-dose NAC. Outcomes included safety and clinical parameters, inflammatory (total leukocyte numbers, cell differentials, TNF-α, IL-8) measures in induced sputum, and concentrations of extracellular glutathione in induced sputum and blood.

Results

High-dose NAC was a well-tolerated and safe medication. High-dose NAC did not alter clinical or inflammatory parameters. However, extracellular glutathione in induced sputum tended to increase on high-dose NAC.

Conclusions

High-dose NAC is a well-tolerated and safe medication for a prolonged therapy of patients with CF with a potential to increase extracellular glutathione in CF airways.     

Cyclooxygenase-2 plays an essential part in cardioprotection of delayed phase of recombinant human erythropoietin preconditioning in rats

Aims

To study the cardioprotection of recombinant human erythropoietin (rhEPO) preconditioning (EPC) and to investigate the role and possible mechanism of cyclooxygenase (COX)‐2 in the delayed phase of EPC.

Methods

In phase 1, myocardial ischaemia reperfusion (I‐R) rat model was established by 30 minutes ligation of left descending coronary and three hours of reperfusion. Rats were given 0.9% saline solution or rhEPO 24 hours before I‐R protocol. COX‐2 selective inhibitor celecoxib was given for further investigation of the cardioprotection of EPC. At the end of I‐R protocol, infarct sizes were measured and ultrastructural organisations were studied. In phase 2, myocardial COX‐2 mRNA expressions and prostaglandins (PGs) contents were studied in different groups after euthanasia.

Results

It was found that EPC could elicit potent cardioprotection against I‐R injury, shown by reduction of infarct size and improvement of ultrastructural organisation; whereas administration of celecoxib resulted in complete loss of this protection. EPC resulted in robust increase in COX‐2 mRNA and PGs levels that were also abrogated by celecoxib.

Conclusions

COX‐2 plays an essential part in cardioprotection of the delayed phase of EPC in rats, which might be related to actions of PGE₂ or PGI₂, or both.     

Twenty-seven years follow-up of a patient with congenital retinocephalofacial vascular malformation syndrome and additional congenital malformations (Bonnet-Dechaume-Blanc syndrome or Wyburn-Mason syndrome)

Purpose

Follow-up of vascular changes in a patient with congenital retinocephalofacial vascular malformation syndrome.

Methods

MRI and cerebral angiography.

Results

In a 36-year-old man, magnetic resonance im aging of the skull and cerebral angiography revealed left intracranial arteriovenous malformations. Follow-up observation of 27 years revealed no essential change of retinal and cerebral arteriovenous malformations. Additional congenital deficits in this patient were described.

Conclusion

Patients with retinal arteriovenous malformations should be early examined with neuroradiological methods.     

Inhibition of quorum sensing in Chromobacterium violaceum by Syzygium cumini L. and Pimenta dioica L.

Objective

To investigated into the anti-quorum sensing (QS) activity of Syzygium cumini L. (S. cumini) and Pimenta dioica L. (P. dioica) using Chromobacterium violaceum (C. violaceum) strains.

Methods

In this study, anti-QS activity of ethanol extract of Syzygium cumini L. and Pimenta dioica L. were screened using C. violaceum CV026 biosensor bioassay. By bioassay guided fractionation of S. cumini and P. dioica, ethyl acetate fraction (EAF) with strong anti-QS activity was separated. Inhibition of QS regulated violacein production in C. violaceum ATCC12472 by EAF was assessed at different concentrations. The effect of EAF on the synthesis of autoinducer like N-acyl homoserine lactone (AHL) was studied in C. violaceum ATCC31532 using its mutant C. violaceum CV026 by standard methods.

Results

EAF inhibited violacein production in C. violaceum ATCC12472 in a concentration dependent manner without significant reduction in bacterial growth. Complete inhibition of violacein production was evidenced in 0.75-1.0 mg/mL concentration of EAF without inhibiting the synthesis of the AHL. TLC biosensor overlay profile of EAF revealed two translucent spots in S. cumini and P. dioica that inhibited C₆-AHL mediated violacein production in C. violaceum CV026.

Conclusions

This study indicates the anti-QS activity of the tested medicinal plants against C. violaceum.     

Screening of biological activities of Polygonum maritimum L. from Algerian coast

Objective

To investigate the antioxidant and the antibacterial activities of crude extract from aerial part of Polygonum maritimum L. (Polygonaceae) (P. maritimum) and to find new actives biomolecules.

Methods

The whole plant was collected from the Rechgoune coast (West of Algeria), and methanolic crude extract of aerial parts of P. maritimum (PMCE) was prepared. The extract was tested against different bacterial strain and tested for his ability to neutralize free radical (DPPH) and to scavenge the H₂O₂.

Results

PMCE had a very high content of total phenol, which was (352.49±18.03) mg/g dry weight, expressed as gallic acid equivalent. PMCE exhibited excellent antioxidant activity, as measured using DPPH and H₂O₂ scavenging assays. It also showed a high antibacterial activity against gram-positive bacterial strains: Bacillus cereus, Bacillus subtilis and Staphylococcus aureus with an highest MIC of 120 µg/mL.

Conclusions

The antioxidant and antibacterial activity of the PMCE is probably due to phenolic compounds present in the extract. The contemporary presence of antioxidant and antibacterial activities in the PMCE suggests that this plant may be a source of bioactive substances with multifaceted activity.     

Research and applications: Patient-level temporal aggregation for text-based asthma status ascertainment

Objective

To specify the problem of patient-level temporal aggregation from clinical text and introduce several probabilistic methods for addressing that problem. The patient-level perspective differs from the prevailing natural language processing (NLP) practice of evaluating at the term, event, sentence, document, or visit level.

Methods

We utilized an existing pediatric asthma cohort with manual annotations. After generating a basic feature set via standard clinical NLP methods, we introduce six methods of aggregating time-distributed features from the document level to the patient level. These aggregation methods are used to classify patients according to their asthma status in two hypothetical settings: retrospective epidemiology and clinical decision support.

Results

In both settings, solid patient classification performance was obtained with machine learning algorithms on a number of evidence aggregation methods, with Sum aggregation obtaining the highest F₁ score of 85.71% on the retrospective epidemiological setting, and a probability density function-based method obtaining the highest F₁ score of 74.63% on the clinical decision support setting. Multiple techniques also estimated the diagnosis date (index date) of asthma with promising accuracy.

Discussion

The clinical decision support setting is a more difficult problem. We rule out some aggregation methods rather than determining the best overall aggregation method, since our preliminary data set represented a practical setting in which manually annotated data were limited.

Conclusion

Results contrasted the strengths of several aggregation algorithms in different settings. Multiple approaches exhibited good patient classification performance, and also predicted the timing of estimates with reasonable accuracy.     

Isolation of a small molecule with anti-MRSA activity from a mangrove symbiont Streptomyces sp. PVRK-1 and its biomedical studies in Zebrafish embryos

Objective

The aim of the present study was to isolate the anti-MRSA (Methicillin Resistant Staphylococcus aureus) molecule from the Mangrove symbiont Streptomyces and its biomedical studies in Zebrafish embryos.

Methods

MRSA was isolated from the pus samples of Colachal hospitals and confirmed by amplification of mecA gene. Anti-MRSA molecule producing strain was identified by 16s rRNA gene sequencing. Anti-MRSA compound production was optimized by Solid State Fermentation (SSF) and the purification of the active molecule was carried out by TLC and RP-HPLC. The inhibitory concentration and LC₅₀ were calculated using Statistical software SPSS. The Biomedical studies including the cardiac assay and organ toxicity assessment were carried out in Zebrafish.

Results

The bioactive anti-MRSA small molecule A₂ was purified by TLC with Rf value of 0.37 with 1.389 retention time at RP-HPLC. The Inhibitory Concentration of the purified molecule A₂ was 30 µg/mL but, the inhibitory concentration of the MRSA in the infected embryo was 32-34 µg/mL for TLC purified molecule A₂ with LC₅₀ mean value was 61.504 µg/mL. Zebrafish toxicity was assessed in 48-60 µg/mL by observing the physiological deformities and the heart beat rates (HBR) of embryos for anti MRSA molecule showed the mean of 41.33-41.67 HBR/15 seconds for 40 µg/mL and control was 42.33-42.67 for 15 seconds which significantly showed that the anti-MRSA molecule A₂ did not affected the HBR.

Conclusions

Anti-MRSA molecule from Streptomyces sp PVRK-1 was isolated and biomedical studies in Zebrafish model assessed that the molecule was non toxic at the minimal inhibitory concentration of MRSA.     

Biochemical characters and antibiotic susceptibility of Staphylococcus aureus isolates

Objective

To observe the biochemical characters and antibiotic susceptibility of isolated Staphylococcus aureus (S. auerus) strains against some conventional and traditional antibiotics.

Methods

Thirty post operative pathogenic isolated S. aureus strains were used in this study. Bacterial culture was done in Mueller-Hinton broth at 37 °C. Characters of these strains were determined by traditional biochemical tests such as hydrolysis test of gelatin, urea, galactose, starch and protein, and fermentation of lactose and sucrose. Antibiotic susceptibility were carried out by minimum inhibitory concentration test, minium bactericidal concentration test, disc agar diffusion test and brain heart infusion oxacillin screening agar.

Results

From this study, it was observed that 100% S. aureus isolates showed positive results in gelatin, urea and galactose hydrolysis test, 50% isolates were positive in starch hydrolysis test, 35% in protein hydrolysis test, 100% isolates in lactose fermenting test, but no isolate was positive in sucrose fermenting test. Antibiotic susceptibility testing suggested that 20% of isolates were resistant to kanamycin and 46.67% were resistant to oxacillin.

Conclusions

These findings show that all these isolates have gelatin, urea, galactose hydrolysis and lactose fermenting activity. 20% of these isolates were resistant to kanamycin and 46.67% were resistant to oxacillin.     

Evaluation of anti-apoptotic activity of different dietary antioxidants in renal cell carcinoma against hydrogen peroxide

Objective

To evaluate the anti-apoptotic and radical scavenging activities of dietary phenolics, namely ascorbic acid,α-tocopherol acetate, citric acid, salicylic acid, and estimate H₂O₂-induced apoptosis in renal cell carcinoma cells.

Methods

The intracellular antioxidant potency of antioxidants was investigated. H₂O₂-induced apoptosis in RCC-26 was assayed with the following parameters: cell viability (% apoptosis), nucleosomal damage and DNA fragmentation, bcl-2 levels and flow cytometery analysis (ROS production evaluation).

Results

The anticancer properties of antioxidants such as ascorbic acid, α-tocopherol acetate, citric acid, salicylic acid with perdurable responses were investigated. It was observed that these antioxidants had protective effect (anti-apoptotic activity) against hydrogen peroxide (H₂O₂) in renal cell carcinoma (RCC-26) cell line.

Conclusions

This study reveals and proves the anticancer properties. However, in cancer cell lines anti-apoptotic activity can indirectly reflect the cancer promoter activity through radicals scavenging, and significantly protect nucleus and bcl-2.     

Tamoxifen Therapy for Breast Cancer and Endometrial Pathology

Background

Tamoxifen, used as adjuvant therapy for carcinoma breast in postmenopausal women to prevent relapse has estrogenic effect on the endometrium.

Methods

104 patients on tamoxifen for more than six months were subjected to a clinical examination and transvaginal sonography. Patients with endometrial thickness > 8 mm were further evaluated by hysteroscopy and endometrial biopsy.

Results

35(34%) patients were symptomatic. The average endometrial thickness was 11.2 mm which correlated with duration of tamoxifen use. 27(48%) patients had abnormal hysteroscopic findings. 35 (63%) of endometrial biopsies revealed abnormal endometrium. One case of endometrial carcinoma was diagnosed. The results were statistically analysed. There is a significant association between symptomatic status and endometrial thickness and duration of tamoxifen use.

Conclusion

All patients on long term tamoxifen should be annually screened for endometrial pathology.Key Words: Tamoxifen, Endometrium, Transvaginal sonography     

African eggplant (Solanum anguivi Lam.) fruit with bioactive polyphenolic compounds exerts in vitro antioxidant properties and inhibits Ca2+-induced mitochondrial swelling

Objective

To evaluate the antioxidant and radical scavenging activities of Solanum anguivi fruit (SAG) and its possible effect on mitochondrial permeability transition pore as well as mitochondrial membrane potential (ΔΨ_m) isolated from rat liver.

Methods

Antioxidant activity of SAG was assayed by using 2,2-diphenyl-1-picrylhydrazyl (DPPH), reducing power, iron chelation and ability to inhibit lipid peroxidation in both liver and brain homogenate of rats. Also, the effect of SAG on mitochondrial membrane potential and mitochondrial swelling were determined. Identification and quantification of bioactive polyphenolics was done by HPLC-DAD.

Results

SAG exhibited potent and concentration dependent free radical-scavenging activity (IC₅₀/DPPH=275.03±7.8 µg/mL). Reductive and iron chelation abilities also increase with increase in SAG concentration. SAG also inhibited peroxidation of cerebral and hepatic lipids subjected to iron oxidative assault. SAG protected against Ca²⁺ (110 µmol/L)-induced mitochondrial swelling and maintained the ΔΨ_m. HPLC analysis revealed the presence of gallic acid [(17.54±0.04) mg/g], chlorogenic acid (21.90±0.02 mg/g), caffeic acid (16.64±0.01 mg/g), rutin [(14.71±0.03) mg/g] and quercetin [(7.39±0.05) mg/g].

Conclusions

These effects could be attributed to the bioactive polyphenolic compounds present in the extract. Our results suggest that SAG extract is a potential source of natural antioxidants that may be used not only in pharmaceutical and food industry but also in the treatment of diseases associated with oxidative stress.     

Finding falls in ambulatory care clinical documents using statistical text mining

Objective

To determine how well statistical text mining (STM) models can identify falls within clinical text associated with an ambulatory encounter.

Materials and Methods

2241 patients were selected with a fall-related ICD-9-CM E-code or matched injury diagnosis code while being treated as an outpatient at one of four sites within the Veterans Health Administration. All clinical documents within a 48-h window of the recorded E-code or injury diagnosis code for each patient were obtained (n=26 010; 611 distinct document titles) and annotated for falls. Logistic regression, support vector machine, and cost-sensitive support vector machine (SVM-cost) models were trained on a stratified sample of 70% of documents from one location (dataset A_train) and then applied to the remaining unseen documents (datasets A_test–D).

Results

All three STM models obtained area under the receiver operating characteristic curve (AUC) scores above 0.950 on the four test datasets (A_test–D). The SVM-cost model obtained the highest AUC scores, ranging from 0.953 to 0.978. The SVM-cost model also achieved F-measure values ranging from 0.745 to 0.853, sensitivity from 0.890 to 0.931, and specificity from 0.877 to 0.944.

Discussion

The STM models performed well across a large heterogeneous collection of document titles. In addition, the models also generalized across other sites, including a traditionally bilingual site that had distinctly different grammatical patterns.

Conclusions

The results of this study suggest STM-based models have the potential to improve surveillance of falls. Furthermore, the encouraging evidence shown here that STM is a robust technique for mining clinical documents bodes well for other surveillance-related topics.     

Foot Protective System : A Boon to Demining Soldiers

Background

A significant number of soldiers suffer serious injuries or die in the process of demining and a protective device is necessary in such operations.

Methods

All the patients involved in demining operations using the foot protection devices in last one year were analysed.

Results

We received 35 patients, of which only five had limb injuries and 30 had eye injuries. Of the five, three had closed calcaneal fractures and two contusion of foot. No fatality, amputation or compound fracture occured.

Conclusion

Early results after the use of this device during demining operations are encouraging.Key Words: Mine blast injury, Demining, Spider boot