MANAGEMENT of ACUTE ALUMINUM PHOSPHIDE TOXICITY in RAT MODEL with a NOVEL INTERVENTION, a TRIAL of BORIC ACID *Ola Abdel Hady Sweilum, *Fatma Shaban Kandeel

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MANAGEMENT OF ACUTE ALUMINUM PHOSPHIDE TOXICITY IN RAT MODEL WITH A NOVEL INTERVENTION, A TRIAL OF BORIC ACID *Ola Abdel Hady Sweilum, *Fatma Shaban Kandeel. **Dalia Abdel Razik Noya *Forensic Medicine and Clinical Toxicology Department, Faculty of Medicine - Menoufia University. **Histology Department, Faculty of Medicine - Menoufia University

ABSTRACT Aluminum phosphide (AlP) or rice tablet is a cheap pesticide. When it comes in contact with acid (gastric acid) or moisture, it releases phosphine (PH3) gas. The heart, lungs, liver are the main targets in acute Aluminum phosphide (AlP) poisoning. Most deaths occur due to cardiovascular toxicity. Recently, boric acid has been proposed theoretically as an antidote for AlP poisoning. With the formula B (OH)3, boric acid can accept electrons as it is a Lewis acid with an empty p orbital. The aim of the present study was to investigate the possible protective role of boric acid in management of aluminum phosphide poisoning and its possible use as an effective antidote to reduce ALP poisoning mortality. Forty albino rats, all of which were adults and males were equally divided into four groups, group 1: control (given distilled water), group 2: given boric acid only (100mg/kg nontoxic dose), group 3: given aluminum phosphide (12.5 mg/kg), group 4: given aluminum phosphide followed by boric acid. Rats in group 1 and 2 was killed by cervical dislocation, while rats in group 3 and 4 were left to die, then all were dissected out. Fixation in situ was performed by perfusion technique then the chest and the abdomen were carefully incised to reveal the heart and the liver. The liver and the heart were washed with saline then quickly dissected, excised then blotted for histological preparation. Blood samples were collected from each rat (immediately after death) for measuring liver enzymes (SGOT, SGPT, ALK (alkaline phosphatase) and cardiac troponin. Rats treated with ALP followed by boric acid showed significant increase in their survival time, marked improvement in histopathological changes and also showed significant improvement in biochemical parameters including liver and cardiac markers (SGOT, SGPT, and cardiac troponin) compared with rats intoxicated with AlP. Conclusion: This study may increase the hope of using boric acid as a new effective antidote for ALP poisoning and opens the door of opportunity for larger experimental and then clinical trials to reevaluate the role of boric acid in acute ALP poisoning. Key words: Aluminum phosphide, Boric acid, Toxicity, Cardiac troponin, Antidote.

INTRODUCTION (PH3); a colorless, inflammable, water Aluminum phosphide (AlP) is a solid insoluble, and highly toxic gas cheap fumigant pesticide (rice tablet) (Moghadamnia 2012; Bumbrah et al., which protects stored grains from 2012). While odorless in its pure form, insects and rodents with a fatal dose PH3 can smell of garlic or decaying fish between 0.15 and 0.5 grams (0.0053 due to the presence of impurities such and 0.0176 oz) (Wahab et al., 2008). as substituted phosphines and When it comes into contact with diphosphines (Wahab et al., 2009). moisture or acid, it releases phosphine

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Phosphine gas (PH3) is a Lewis humans in nutritional amounts, and is base and a strong nucleophile. It is a thus found in animal and human tissues reducing agent with a lone-pair electron at low concentrations (Environmental which reduces cytochrome c oxidase Protection Agency 2004). It is and interferes with the electron transfer considered as a weak acid and it is non- from complex III to complex IV of the toxic in rats with a median lethal oral mitochondrial respiratory chain, dose of 5.14 g kg-1 (Perelygin and ultimately resulting in the inhibition of Chistyakov 2006). oxidative phosphorylation, adenosine In the study done by Soltani et al., triphosphate depletion and cell death (2016), at Sultan Qaboos University on (Soltani et al., 2013). chemical reaction between boric acid Aluminum phosphide is usually and phosphine, they concluded that taken in suicidal attempt. It was boric acid significantly reduced the rate reported that AlP has a mortality rate of phosphine gas evolution (P <0.01) more than 50% of intoxicated cases and that boric acid may be an efficient (Moghadamnia 2012). After ingestion and non-toxic antidote for PH3 of aluminum phosphide, phosphine gas poisoning. is released in the stomach and after absorption; symptoms and signs of AIM OF THE STUDY poisoning occur rapidly (Wahab et al., This study aimed to investigate the 2008) which include nausea, vomiting, possible antidotal role of boric acid in garlic breath smell, retrosternal and management of Aluminum phosphide epigastric pain, anxiety, agitation, and toxicity and its protective role on Alp- dyspnea (Mostafazadeh 2012 & Popp induced biochemical and histo- et al 2002). pathological changes in liver and heart The heart, lungs and liver are the of adult male albino rats. main target organs in acute AlP poisoning, although multi-organ system MATERIALS & METHODS failure may also occur. Most deaths This experiment was carried out in occur within the first 12-24 hours, the animal laboratory in Faculty of usually because of cardiovascular Medicine, Menoufia University. toxicity (Perry 2007). Animals: Forty adult male albino Patients are usually managed rats weighed 150-240 grams were used conservatively, and their outcome is in this experiment. Animals were very bad as there is no efficient antidote purchased from a small animal breeding has been discovered for AlP poisoning center in Shebin El-kom, Menoufia, till now (Moghadamnia 2012). Egypt. All ethical protocols for animal Recently, boric acid has been treatment were followed. This proposed theoretically as an antidote for experimental protocol was accepted by AlP poisoning. With the formula B the Ethical Committee of Menoufia (OH)3, boric acid is a Lewis acid which faculty of medicine. can accept electrons as it has an empty Materials: Aluminum phosphide p orbital (Soltani et al., 2013). Boric used in experiment was in the form of 3 acid is a colorless, odorless, transparent gm tablets (57% concentration, Excel material used in industrial sector for crop care Ltd, India), and contains two preparation of disinfectants and drugs. compounds: aluminium phosphide and However, it is primarily beneficial for Egypt J. Forensic Sci. Appli. Toxicol Vol 17 (2) December 2017 Sweilum et al. 59 aluminium carbonate in a ratio of the abdomen were carefully incised to 57:43. reveal the heart and the liver. The heart Aluminum phosphide Purchased and the liver were washed with saline from local pesticide shop in Shebin El- then quickly dissected, excised then Kom. blotted. Boric acid was obtained from I- Histological study: Tissue toxicology laboratory in toxicology samples were fixed in 10% formol department (boric acid powder saline for 2-5 days then washed in tap dissolved in hot distilled water and water, dehydrated in ascending grades given in a dose of 100 mg/kg dose), of alcohol (50%, 70%, 90% over night Menoufia Faculty of medicine. and 100% for 3 hours), cleared in Substances of experiment were given xylene and embedded in paraffin wax. by gavage (oral tube). Paraffin sections of 5-6 micrometer Experimental design: thickness were cut and stained with Rats were housed in polypropylene hematoxylin & eosin (H &E). cages. Food and water were provided II- Immunohistochemical study: ad libitum (on desired). Rats were given Caspase-3: sections were stained with two days’ time to get acclimatized with the primary rabbit polyclonal anti the laboratory conditions prior to caspase-3 antibody (Thermo Scientific, experiment. All rats were fasted for 24 Lab Vision, USA) (Sanii et al., 2012). hours just before starting the Immunostaining by caspas3 show if experiment (in order to empty gastro there is apoptosis or not, as we want to intestinal tract and abolishing know if ALP produce effect at the level interactions of food components with of the nucleus or not. the tested substance) (vermeulen et al., Biochemical analysis: 1997). Blood samples were collected from Rats were divided into four groups, each rat (immediately after death) for each containing ten rats (n=10). measuring liver enzymes (SGOT, Group 1: Control (given distilled SGPT, ALK (alkaline phosphatase) and water) cardiac troponin. Blood was centrifuged Group 2: Boric acid only (+ve at 3000 rpm to obtain the plasma, then control) in a dose, (100 mg/kg nontoxic SGOT, SGPT activity were determined dose), (Ince et al., 2010). (Ritman and Frankel 1957). Activity Group 3: Given aluminum of alkaline phosphatase was determined phosphide (12.5 mg/kg) (Baeeri et al., (Babson and Read 1959), and the 2013). levels of cardiac troponin were also Group 4: Given aluminum measured by enzyme linked phosphide (12.5 mg/kg) followed by immunoassay (ELISA) technique using boric acid (100mg/kg). a standard kit (Glory Science Co., Ltd, Rats in group 1 and 2 was killed by USA). cervical dislocation, while rats in group Statistical analysis: 3 and 4 were left to die, then all were The collected data were organized, dissected out. tabulated and analyzed using SPSS Histological specimen version 16 software (Spss Inc, Chicago, preparation: ILL Company). The t-test was used to Fixation in situ was performed by compare between mean ± SD of two perfusion technique then the chest and groups. Also, normally quantitative Egypt J. Forensic Sci. Appli. Toxicol Vol 17 (2) December 2017 Sweilum et al. 60 data expressed in mean ± SD and was Sections of the liver of rats compared using F test (ANOVA), and intoxicated with Aluminium phosphide the Sig. between groups was done using (group 3) showed destructive changes Post Hoc Test (Tukey). The abnormally in the form of hydropic degeneration of distributed quantitative data was hepatocytes, presence of many expressed in median (Min. - Max.), and hepatocytes with pyknotic nuclei (Fig.3 compared using Kruskal Wallis test, A1). Many vacuoles were also seen where Sig. between groups was done among hepatic tissue. Dilated distorted using Post Hoc Test (Dunn's multiple and congested blood sinusoids were comparisons test). The accepted level noticed (Fig.3 A2). Central vein of significance was stated ≤ 0.05. showed dilation and congestion. Mononuclear cells infiltrated the area RESULTS around central vein and blood sinusoids A- Histopathological & (Fig.3 A3). Immunostaining for immunoreactivity changes: Caspase 3 showed + ve reaction  Liver sections (Fig.3C). Sections of the control group Liver sections in rats intoxicated (group 1) showed hepatocytes arranged with aluminum phosphide then treated in plates radiating from central vein and with boric acid (group 4) showed separated by blood sinusoids. The marked improvement of the hepatocytes characterized by vesicular degenerative changes as the central nuclei and acidophilic hepatocytes showed acidophilic cytoplasm (Fig.1 A). Immunostaining cytoplasm with central vesicular nuclei for Caspas 3 showed –ve reaction (Fig.4 A1). There was marked decrease (Fig.1 C). in the presence of vacuoles. The blood Sections of the boric acid treated sinusoids showed normal appearance. group (group 2) showed picture of liver The disappearance of monocells similar to control group which appeared infiltrates was also noticed (Fig.4 A2). completely normal (Fig. 2 A&C). Immunstaining by Caspase 3 showed – ve reaction (Fig. 4 C).

Figure (1A): A photomicrograph in a Figure (1C): photomicrograph in a section of control rat liver showing section of control rat liver showing -ve hepatocytes (►) arranged in plates that Caspas 3 reaction. x 400. radiate from the central vein (C), and separated by blood sinusoids (S). H. and E. x 400

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Figure (2A): A photomicrograph in a Figure (2C): A photomicrograph in a section of rat liver treated with Boric acid section of rat liver treated with Boric acid showing normal hepatocytes (►) showing –ve Caspas 3 reaction x 400. radiating from central vein (C) and separated by blood sinusoids (S) as control group. H. and E. x 400.

Figure (3A2): A photomicrograph in a Figure (3A2): A photomicrograph in a section of rat liver treated with aluminum section of rat liver treated with aluminum phosphide showing distorted, dilated, phosphide showing cytoplasmic vacular congested blood sinusoids (→), and many degeneration (↔), pykontic nuclei (P) in cytoplasmic vacuoles (V) were noticed. the affected hepatocytes and dilated Some hepatocytes showing hydropic congested central vein (Cg). H. & E. x degeneration (D) and pykontic nuclei (P). 400. H. & E. x 400.

Figure (3A): A photomicrograph in a Figure (3C): A photomicrograph in a section of rat liver treated with aluminum section of rat liver treated with aluminum phosphide showing monocellular phosphide showing strong +ve Caspas infiltration (ↄ). Cytoplasmic vacuoles (↔) reaction. X 400. were seen. Hydropic degeneration of hepatocytes (D) and pykontic nuclei (P) were also noticed H. & E. x 400 Egypt J. Forensic Sci. Appli. Toxicol Vol 17 (2) December 2017 Sweilum et al. 62

Figure (4A1): A photomicrograph in a Figure (4A2) A photomicrograph in a section of rat liver treated with section of rat liver treated with aluminum aluminum phosphide and Boric acid phosphide and Boric acid showing return of showing normally arranged hepatocytes. liver tissue to its normal appearance. Only few blood sinusoids are dilated Improvement of sinusoidal dilation (→), (→) absence of monocellular infiltration but x400 few vacuoles are still present (V) H. & E. x 400

Figure (4C): A photomicrograph in a section of rat liver treated with aluminum phosphide and Boric acid showing –ve Caspas reaction. X 400

 Heart sections fibers of the heart, many cytoplasmic In control group (group 1), Cardiac vacuoles and hyaline materials were myocytes appeared cylindrical with noticed. Also, pyknotic nuclei were acidophilic striated sarcoplasm and present in the affected fibers (Fig.7 centrally located oval nuclei. A1& A2). Widening of interstitium Fibroblasts with its deeply stained between cardiac myocytes was noticed, flattened nuclei were noticed at the C.T which contained hemorrhage and (connective tissue) endomysium (Fig. infiltration with mononuclear cells. 5A). Immunostaining for Caspase 3 There was congestion of blood vessels showed –ve reaction (Fig. 5 C). (Fig.7 A3). Caspase 3 immunostaining In group 2 (Boric acid treated), revealed strong +ve reaction (Fig.7 C). examination by light microscope In group 4 (Aluminum phosphide showed cardiac myocytes with and Boric acid treated), examination by acidophilic striated sarcoplasm and light microscopic showed appearance centrally located elongated vesicular of cardiac myocytes more or less nuclei, nearly as picture in control similar to control group (Fig.8 A1 & group (Fig. 6 A & C). A2). Also Caspase 3 Immunostaining Microscopic changes noticed in showed -ve reaction (Fig.8 C). group 3 (Aluminum phosphide) were in the form of fragmentation of muscle Egypt J. Forensic Sci. Appli. Toxicol Vol 17 (2) December 2017 Sweilum et al. 63

Figure (5A): A photomicrograph in a Figure (5C): A photomicrograph in a section of control rat heart showing long section of control rat heart showing -ve cylindrical cardiac muscle fiber (C) with Caspas 3 reaction. x 400 acidophilic striated sarcoplasm and centrally located oval nuclei. The cardiac muscle fibers are connected by intercalated discs (IC). Flattened deeply stained nuclei of fibroblast are noticed (F). H.& E. x 1000

Figure (6A): A photomicrograph in a Figure (6C): A photomicrograph in a section of rat heart treated with Boric acid section of rat heart treated with Boric acid showing cardiac muscle fibers with showing –ve Caspas 3 reaction. X400 acidophilic sarcoplasm (C) and central vesicular nuclei similar to control group. H. &E. x 1000

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Figure (7A1): A photomicrograph in a Figure (7A2): A photomicrograph in a section of rat heart treated with aluminum section of rat heart treated with aluminum phosphide showing fragmentation of phosphide showing marked hyalinization cardiac muscle fibers (→), cytoplasmic (↔) and cytoplasmic vacuolation (V). vacuolation (V) and hyaline material Widening of the interstitial space accumulation (↔). Notice the pykontic separating the cardiac myocytes (►), nuclei (P) in some cardiac myocytes. H.& containing hemorrhage and mononuclear E. x 1000 cells infiltrate. H. & E. x 1000

Figure (7A3): A photomicrograph in a Figure (7C) A photomicrograph in a section of rat heart treated with aluminum section of rat heart treated with aluminum phosphide showing marked interstitial phosphide showing +ve Caspas 3 reaction. edema (►), congested blood vessels are X 400 also seen (Cg). H. & E. x 1000

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Figure (8A1): A photomicrograph in a Figure (8A2): A photomicrograph in a section of rat heart treated with aluminum section of rat heart treated with aluminum phosphide and Boric acid showing normal phosphide and Boric acid showing structure of cardiac myocytes (c) with its minimal interstitial edema. The cardiac centrally located oval vesicular nuclei. muscle fibers regain its normal shape and Few vacuoles are still present (V).H. & E. structure. H. & E. x 1000 X 1000

Figure (8C): A photomicrograph in a section of rat heart treated with aluminum phosphide and Boric acid showing –ve Caspas 3 reaction. x 400.

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B- Biochemical results: Liver enzymes, and cardiac troponin:

Table (1): Comparison between the different studied groups according to liver enzymes (SGOT, SGPT, ALK) and cardiac troponin. Control Boric ALP ALP+boric p (n= 10) (n= 10) (n= 10) (n= 10) SGOT 47 ± 45.5 ± 68.7ab ± 51.9abc ± <0.001* U/L 2.5 2.3 1.5 2.4 SGPT 36.6 ± 36.2 ± 75.6ab ± 61.3abc ± <0.001* U/L 2.5 2.8 7.8 5.6 ALK 319 325 490ab 450ab <0.001* U/L (313–420) (300–420) (444–570) (430–500) Troponin 0.03 0.03 0.26 ab 0.06 abc <0.001* Ug/L (0.02–0.04) (0.02–0.05) (0.09–0.40) (0.03–0.20) Normally quantitative data was was done using Post Hoc Test (Dunn's expressed in mean ± SD and was multiple comparisons test) compared using F test (ANOVA), Sig. a: significant with control bet. grps was done using Post Hoc Test b: significant with boric (Tukey) abnormally distributed c: significant with ALP quantitative data was expressed in *: Statistically significant at p ≤ 0.05 median (Min. - Max.) and was compared using Kruskal Wallis test, Sig. bet. grps

Data of the current work showed ALP) there was a significant decline in that group of Alp poisoned rats showed liver enzymes activity (SGOT, SGPT) a highly significant increase in liver compared with ALP group. Concerning enzyme activity and cardiac troponin cardiac troponin, its level was compared to their levels in rats of either significantly decreased compared with control or boric acid groups (P value its level in group 4 (Boric acid and ALP <0.001). In group 4 (Boric acid and group) (Table 1).

Table (2): Survival time in mean ± SD of rats intoxicated with ALP compared with that treated with boric acid with ALP. ALP group ALP + Boric t - value P value acid Survival time 98.6 (±7.06) 511.3 (±11.84) 94.67 < 0.0001 in minutes Using t- test Statistically significant at p ≤ 0.05

Mean survival time was 98.6 min <0.0001) (Table 2). (±7.06) after a single dose of aluminum phosphide of 12.5mg/kg. Increased to DISCUSSION 511.3min (±11.84) in AlP group treated Acute intoxication with phosphides with boric acid (group4) with much is a serious health problem either increase in survival time and this accidental or suicidal in Menoufia increase was highly significant (P value governorate, Egypt as in 2016, at

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Menoufia poison control center /antioxidant balance and it inhibits lipid (MPCC), phosphides were the cause of peroxidation (Turkez et al., 2011). about 80 % of mortality of poisoned These pathological changes due to cases. phosphine gas effects liberated after Until 2012, the main line of ALP ingestion and cause inhibition of management for ALP intoxicated respiration mitochondria in rat liver patients was just supportive due to (Okolie et al., 2004). absence of proper antidote In India, Saraf et al. (2015), (Mostafazadeh, and Farzaneh 2012). reported that zinc phosphide is the most Boric acid is considered as a safe common cause of drug/toxin-induced product and its toxicity is unusual human irreversible acute liver failure. (Kaymaz 2015), so we tried in our The histopathological results of our study to test the possible use of this research are in agreement with the substance as an antidote for ALP experimental study done by Sinha et poisoning in rat models, based on the al. (2003), who recorded theoretical study of Soltani et al., histopathological changes in the rats (2016). liver after ALP intoxication in the form In the current study, the mean of congestion in central veins, survival time was 98.6 (±7.06) after a haemorrhage degeneration, , single dose of aluminum phosphide of hyperplasia in Kupffer cell, sinusoidal 12.5mg/kg. That increased to 511.3 dilation, mononuclear cells infiltration, (±11.84) in AlP group treated with centrilobular necrosis and fatty boric acid. changes. There was much increase in Mehrpour et al. (2010), recorded survival time with administration of liver tissues changes with aluminum boric acid after aluminum phosphide phosphide, in the form of mononuclear which entail that this substance (boric infiltration, congestion of veins and acid) may be the missed antidote for degeneration of hepatocytes. ALP poisoning specially if we compare Musshoff et al. (2008), found it with other proposed antidotes like N - congestion, centrizonal necrosis, edema acetyl cysteine (138 ± 13min with NAC of liver tissue after ALP poisoning. dose=50mg/kg) or vitamin C (250 ± While Saleki et al. (2007) found fine 17min with vitamin C dose= cytoplasmic vacuolization of 500mg/kg) (Moghadamnia, 2000) hepatocytes and sinusoidal congestion Histopathology & were observed after only one-day immunoreactivity of the liver. phosphine gas poisoning. There were histopathological Moghaddamnia et al., (2000), changes in the rat liver tissues in the showed hepatic congestion and liver form of disorganization of hepatocytes, necrosis with ALP poisoning at hydropic degeneration, sinusoidal different doses. dilation and some hepatocytes showed Pykontic nuclei in some pyknotic nuclei in ALP group in hepatocytes of ALP group were noticed comparison with control and boric acid in the current study. This finding was only groups. These changes were confirmed with the apparent increase in markedly decreased when rats were Caspase 3 immunoreactivity in liver treated with boric acid after ALP. As sections. This could be explained by boric acid act to maintain the oxidant DNA damage and inhibition of DNA Egypt J. Forensic Sci. Appli. Toxicol Vol 17 (2) December 2017 Sweilum et al. 68 synthesis. myoctyloysis and degeneration which Hydropic degeneration and suggestive of myocardial injury, and pykontic changes of hepatocytes were they also found areas of increased also reported by Olurin et al. (2016). waviness of myocardial fibers that Heart histopathology: indicate an episode of myocardial Heart examination of ALP rat infarction. group revealed fragmentation of muscle Rahbar et al. (2010) revealed fibers, cytoplasmic vacuolation and congestion, leucocytes infiltration and hyalinzation, widening of interstitium necrosis in myocardial tissue in their between cardiac myocytes, hemorrhage, study on aluminum phosphide. mononuclear cellular infiltration and Biochemical changes. congestion of blood vessels compared As regards data of the current work, with that of boric acid (only) and the activities of liver enzymes SGOT control groups. These changes were and SGPT, alkaline phosphatase (AlK) markedly decreased in ALP and boric and cardiac troponin in plasma of ALP acid group. intoxicated rats showed a significant These effects on heart tissue were increase compared with its activity in due to Phosphine gas liberated after rats groups of control and boric acid ALP ingestion which induces only; these elevations were decreased in myocardial contractility impairment group 4 (ALP and boric acid) which and leads to circulatory failure and showed a significant decrease as pulmonary edema (Sogut et al, 2011). regards liver enzymes (SGOT and Many pykontic nuclei were seen in SGPT) and cardiac troponin. the affected cardiac muscle fibers. This These biochemical results were in was confirmed by an apparent increase agreement with Hamdan and Bonin of Caspase 3 immunoreactivity in ALP (1999), as they also recorded increased treated rats. Some investigators stated serum alkaline-phosphatase (ALk) that a strong relation is found between activity in rats after exposure to heart tissue injury and cellular hypoxia phosphine gas. Significant increasing in - induced mitochondrial dysfunction activity of ALT with ALP toxicity also following ALP poisoning (Karamaz recorded by Morgan et al., (1995). et. at., 2013). In the current study there was also a Boric acid help protection of significant decrease in cardiac troponin nucleic acids from damage by free in "AlP and boric acid" group radicals results in ALP exposure. This compared with AlP group, which explains our result of -ve Caspase 3 means that boric acid carry a beneficial immunoreactivity in rats of ALP treated effect in case of AlP poisoning. with boric acid. (Turkez et al., 2011). El Hangouche et al., (2017), found In accordance to our results, Arora elevated cardiac troponin level in et al. (1995) also observed human patients intoxicated with ALP. histopathological changes in various Myocardial injury with elevated organs (e.g. heart) in the form of cardiac biomarkers (CPK, CK-MB, and congestion, leucocytic infiltration and Troponin-T) was also noticed in the edema in ALP. work of Soltaninejad et al. (2012). Shah et al. (2009), recorded myocardial toxic effects of ALP in the form of myocyte vacuolation and Egypt J. Forensic Sci. Appli. Toxicol Vol 17 (2) December 2017 Sweilum et al. 69

CONCLUSION toxicity of aluminum phosphide. Rats given boric acid after ALP Toxicology and Industrial Health; showed increase in their survival time, 29(2):126-35. marked improvement in the Bumbrah, GS.; Krishan, K.; histopathological and biochemical Kanchan, T. et al. (2012): parameters including liver and cardiac Phosphide poisoning: A review of markers compared with rats given only literature. Forensic Sci Int; 214:1– ALP. 6. So this study opens the door of EL Hangouche, A.J.; Fennich, H. ; opportunity for larger experimental and Alaika, O. et al. (2017): Reversible then clinical trials to reevaluate whether Myocardial Injury and the early use of boric acid in ALP Intraventricular Thrombus poisoning will improve outcome in this Associated with Aluminium highly fatal poisoning or not. Also, it Phosphide Poisoning. Case Rep may increase the hope of using boric Cardiol. acid as a new effective antidote in ALP Environmental Protection Agency poisoned cases. (2004): Toxiclogical review of boron and compounds. Integrated RECOMMENDATIONS: Risk Information System (IRIS) - Further researches needed to Washington DC. study the possible beneficial effect of Hamdan, H. and Bonin, AM. (1999): boric acid in treatment of ALP toxicity Effects of phosphine inhalation on in human. biochemical and genotoxic - Increase awareness about the endpoints in rodents. International danger of phosphide poisoning between Journal of Medical Toxicology and publics especially farmers - Selling of Forensic Medicine. zinc and aluminum phosphide from Ince, S.; Kucukkurt, I.; Cigerci, IH.; pesticide shops should be restricted and et al. (2010): The effect of dietary replaced by other less or nontoxic boric acid and borax forms. supplemantation on lipid - Restriction of availability of these peroxidation, antioxidant activity, highly toxic poisonous materials which and DNA damage in rats. J Trace are used mainly for suicide. Elem Med Biol. 24:161–164. Karami, S.; Jafari, K. and Abdollahi, REFERENCES M. (2013): Comprehensive Review Arora, B.; Punia, RS.; Kalra, R. et al. of the Mechanistic approach and (1995): Histopathological changes related therapies to cardiovascular in aluminium phosphide poisoning. effects of aluminum phosphide. J Indian Med Assoc. 93(10):380-1. International Journal of Babson, AL. and Read, AP. (1959): Pharmacology. Determination of total acid Kaymaz, B.; Gölge, UH.; Ozyalvaclı, phosphatase in serum. Am J Clin G. et al. (2015): Effects of boric Pathol; 32:89–91. acid on the healing of Achilles Baeeri, M.; Shariatpanahi, M.; tendons of rats. Knee Surg Sports Baghaei, A. et al. (2013): On the Traumatol Arthrosc. 24(12):3738- benefit of magnetic magnesium 3744. nanocarrier in cardiovascular Egypt J. Forensic Sci. Appli. Toxicol Vol 17 (2) December 2017 Sweilum et al. 70

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الملخص العربي للبحث عنوان البحث عالج التسمم الحاد لفوسفيد األلومنيوم في نموذج الجرذان بتدخل جديد : تجربه حمض البوريك عال عبد الهادى منصورa, فاطمة شعبان قنديل aداليا عبد الرازق نويهb قسم الطب الشرعى والسموم اإلكلينيكيةa - قسم الهستولوجيb – كلية الطب – جامعة المنوفية مقدمة البحث: فوسفيد األلومنيوم هو من المبيدات الحشريه الرخيصة )قرص األرز(. فعندما يحدث تالمس بينه وبين الرطوبة أو اي حمض يطلق غاز الفوسفين )PH3(. القلب والكبد والرئتين هي األجهزة المستهدفة الرئيسية في التسمم الحاد بفوسفيد االلومنيوم. معظم الوفيات في اآلونة األخيرة، تحدث بسبب سمية القلب واألوعية الدموية. وقد اقترح حمض البوريك نظريا كترياق للتسمم بفوسفيد االلومنيوم. مع الصيغة B (OH)3، حمض البوريك هو حمض لويس مع المداري p التي يمكن أن تقبل اإللكترونات. الهدف من البحث: تهدف هذه الدراسة إلى التحقيق في الدور المحتمل لحمض البوريك كترياق في عالج تسمم فوسفيد األلومنيوم ودورها الوقائي على التغيرات البيوكيميائية والتغيرات النسيجية المرضية في الكبد والقلب في الفئران البيضاء واستخدامها المحتمل كترياق فعال للحد من وفيات التسمم بفوسفيد األلومنيوم. مواد وطرق البحث: أجرى هذا البحث على أربعين من ذكور الجرذان البيضاء و قد قسموا إلى أربع مجموعات كالتالي: المجموعة األولى المجموعة الضابطة السلبية( والتي اعطيت ماء مقطرا بالفم، المجموعة الثانية )الضابطة اإليجابية( التي تم إعطاؤه حمض البوريك فقط، المجموعة الثالثة وتم إعطاؤها فوسفيد االلومنيوم، المجموعة االربعة وتم إعطاؤها فوسفيد االلومنيوم وحمض البوريك بنفس الجرعات السابق وصفها. جميع الجرعات تم إعطاؤها بالفم مره واحده ثم تم التضحية بالجرذان عن طريق خلع فقرة العنق ثم تم التشريح واستئصال الكبد والقلب واالحتفاظ بها في فورمالين لفحصها هيستولوجيا. كما تم اخذ عينات من الدم لتحليل انزيمات الكبد وتروبونين القلب. نتائج البحث: وكانت النتيجة ان خاليا المجموعة الضابطة السلبية والضابطة اإليجابية كانت طبيعية وال يوجد بها أي تغيرات هيستوباثولوجية أو بيوكيميائيه في حين أن هناك تغيرات كبيرة وجدت في خاليا الكبد والقلب للجرذان التي تم إعطاؤها فوسفيد االلومنيوم. كما إن هناك زيادة في معدل إنزيمات الكبد و التروبونين.كثير من هذه التغيرات قلت بصورة ملحوظة في خاليا الجرذان التي تم إعطاؤها حمض البوريك بعد فوسفيد االلومنيوم. االستنتاج: هذه الدراسة تفتح الباب أمام إجراء دراسات تجريبية أكبر ثم تجارب سريرية إلعادة تقييم دور حمض البوريك في عالج التسمم بفوسفيد االلومنيوم. وزيادة األمل في استخدام حمض البوريك باعتباره ترياق فعال جديد.

Egypt J. Forensic Sci. Appli. Toxicol Vol 17 (2) December 2017