The effect of melatonin on ductus epididymis. Unilateral testicular

Brief Communication The effect of melatonin on ductus epididymis. Unilateral testicular torsion in rats Ayse Yildirim, MD, PhD, Murat Akkus, BSc, PhD, Ahmet R. Ersay, MD, Yusuf Nergiz, BSc, PhD, Ozlem P. Baran, MD, PhD.

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he main pathophysiology of testicular torsion is ischemia/reperfusion (I/R) injury of the testis caused by the twisted spermatic cord and its release.1 Although extensive research has been conducted on the pathophysiology of testicular torsion, the effect of ischemia on the epididymis has not been primarily investigated due to the testis and epididymis share a common blood supply (testicular, cremasteric, and vasal vessels).2 The main cause of testicular damage after torsion is oxygen free radicals produced during reperfusion. Oxygen free radicals oxidize membrane lipids, proteins, and DNA, leading to cellular dysfunction and sometimes cell death. Melatonin (N-acetyl-5-methoxytryptamine) is a neurohormone synthesized and secreted predominantly by the pineal gland in the dark and has been shown recently to be a potent free radical scavenger.3,4 This study was designed to investigate the protective effect of melatonin, which is more effective than other free radical scavengers, in I/ R injury after experimental testicular torsion to evaluate the ultrastructural findings of ductus epididymis with or without melatonin in both torsion and detorsion of testis. All procedures complied with the standards for care and use of animal subjects as stated in the Guide for the Care and Use of Laboratory Animals (Institute of Laboratory Animal Resources, Dicle University, DÜSAM). Sprague-Dawley male rats (250-300 g) fed on a standard diet and with tap water ad libitum, in a 12 hour light-dark cycle was used. All animals were anesthetized with an intraperitoneal injection of 50 mg/kg of pentobarbital sodium. Testis I/R injury was induced by torsion of the left testis, with a 720 degree twisting of the spermatic cord so as to produce a total occlusion of testis for 2.5 hours. The same testis was then detorsioned. The Sprague-Dawley male rats were divided into 5 groups, each containing 7 rats. After 2.5 hours of torsion and detorsion, unilateral orchiectomies were performed for histopathological examination. The groups were labeled as first group (control), second group (torsion); third group (torsion plus detorsion), fourth group (torsion plus melatonin 20 mg/kg/IP plus detorsion), fifth group (torsion plus melatonin 50 mg/ kg/IP plus detorsion). For the histological examination, epididymal tissues were fixed in 2.5% glutaraldehyde and postfixation 1% osmic acid solutions, the araldit blocks prepared by routine electron microscopic techniques 288

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were cut in a thickness of 300-400 Aº. They were then examined under transmission electron microscopy after application of contrast stain. Tissue sections were evaluated using a JEOL 1010 transmission electron microscope. The epididymal tissues were intact in controls. In the torsion group, when the microphotographs were examined, the nuclei of basal cells was normal, however, mitochondrial crystolysis and mitochondrial swelling were present in the cytoplasm of columnar cells, dilatation, and sometimes fragmentation in the tubuli of rough endoplasmic reticulum was also detected. Myelin figures were seen in some areas due to cytoplasmic edema. As well as the presence of seconder lysosomes, autophagic vacuoles were noticeable and there was increase in the lipid droplets compared with controls. When the detorsion group was examined, there was an increase of lipid contents in the basal cells, electron dense lipid accumulation, more autophagic vacuoles, increase of seconder lysosomes, not only activated Golgi complex but also mitochondrial crystolysis were seen in the cytoplasm of columnar cells. In some tissue sections of the detorsion group, necrosis had started in the columnar cells, extensive degeneration and increased numbers of autophagic vacuoles in the rough endoplasmic reticulum were also detected. In the melatonin 50 group, electron dense lipid droplets with mitochondrial crystolysis and mitochondrial swelling in the cytoplasm of basal cells were observed. In other sections of this group, there were myelin figures in some areas, and an increase in seconder lysosomes was observed. Therefore, active Golgi complex and lipid accumulation were also noticeable. In the melatonin 20 group, when the microphotographs were investigated, we detected only slight lipid accumulation in the basal cells, the lipid accumulation in the columnar cells was slightly more than the basal cells, and mitochondrial crystolysis with more myelin figures due to the edema of the cytoplasm was also detected. Therefore, the columnar cells had extense cytoplasm and few seconder lysosomes (Figure 1). This study investigated the effects of prolonged ischemia on the ultrastructure of the epididymis and the protective role of melatonin hormone. Kristo et al2 reported that the macroscopic and microscopic observations of their study support the hypothesis that the epididymis is more resistant to ischemia than the testis during testicular torsion. In their study, the group that underwent 4 hours of left testicular and epididymal ischemia showed light microscopic findings, which demonstrated intact epithelia with microvilli, normal nuclear chronicity, and no cellular debris. However, in the same study, the group that underwent 8 hours of left testicular and epididymal ischemia, demonstrated minimal sloughing of the luminal cells in the epididymis. Thus, there was no reported ultrastructural study of epididymal ischemia,

Melatonin, ischemia-reperfusion injury, testis

Figure 1 - Melatonin 20 group demonstrating rat ductus epididymis to which 2.5 hours of torsion performed with administration of 20 mg melatonin. Very few lipid accumulation (LD) compared with the melatonin 50 group in the basal cells (BC), mitochondrial crystolysis (MC) and more myelin figures (MF) seen. Few seconder lysosomes (*) in the columnar cells (CC) has been demonstrated. (Uranyl acetate-lead citrate staining, original magnification x4400).

and when we compared the light microscopic findings of Kristo’s study2 with ours in the torsion group, which underwent 2.5 hours of testicular and epididymal ischemia, we detected mitochondrial swelling and crystolysis in the columnar cells of ductus epididymis. In addition, the presence of seconder lysosomes and autophagic vacuoles was noticeable and there was increase in the lipid droplets. Kristo et al2 considered both the structural and functional differences of the testicular and epididymal response to ischemia.2 In our study, we saw the structural and functional degenerations as Kristo et al had reported, and our observations were similar with theirs. In recent years, several antioxidant agents have been used to prevent I/R-induced tissue damage in experimental testicular torsion such as, superoxide dismutase (SOD), catalase, calcium channel blockers, oxypurinol, and allopurinol, except melatonin.5 Melatonin is a potent antioxidant agent more effective than allopurinol in preventing testicular damage after acute experimental torsion. Even though this is an animal model, melatonin may clinically be used as an antioxidant agent in testicular torsion.6 In our study, we also examined the prophylactic effect of melatonin in 20 and 50 mg/kg doses. Prillaman and Turner5 also studied to determine whether testicular function after one hour of torsion can be rescued by the administration of antioxidant agents (SOD, catalase, verapamil, and allopurinol). These investigators showed that SOD and catalase treatments provide a significant rescue of functioning testicular parenchyma after one hour of torsion, and neither allopurinol nor verapamil adds benefit. However, the dose of allopurinol could have

been inappropriate. No significant rescue was seen in the testes undergoing 2 hours of torsion. In our study, the 2.5 hours of torsion showed similar results to that of Abasiyanik and Dagdonderen.6 The results of melatonin 20 and melatonin 50 were nearest the controls. The prophylactic effect was present, but there was no complete rescue of the tissue. However, Akgur et al7 reported that allopurinol treatment prevents reperfusion injury after testicular torsion lasting as long as 5 hours. In our study, we observed that melatonin 20 was more prophylactic than melatonin 50 according to the results. In Abasiyanik and Dagdonderen study,6 they determined that melatonin treatment prevents I/R injury in testicular tissue both biochemically and histopathologically after 6 hours of torsion.6 In our study, as the melatonin 20 group had little lipid accumulation in the basal cells, and the lipid accumulation in the columnar cells slightly more than the basal cells, we suggest that melatonin 20 mg administration has a protective effect in ischemia of 2.5 hours of the ductus epididymis. In conclusion, after investigation of the ductus epididymis of the rats undergoing ischemia and reperfusion to their testes, the detorsion group had more degeneration than the torsion group. In addition, 20 mg/kg melatonin had a more protective role than 50 mg/kg melatonin on the tissue of the ductus epididymis in damaging testis by ischemia-reperfusion. Received 14th February 2006. Accepted 14th August 2006. From the Department of Histology and Embryology (Yildirim, Akkus, Nergiz, Baran), Faculty of Medicine, Dicle University, Diyarbakir, and the Department of Urology (Ersay), Faculty of Medicine, Onsekiz Mart University, Canakkale, Turkey. Address correspondence and reprint requests to: Dr. Ozlem P. Baran, Department of Histology and Embryology, Faculty of Medicine, Dicle University, 21280 Diyarbakir, Turkey. Tel. +90 (412) 2488001 Ext. 4650. Fax. +90 (412) 2488440. E-mail: [email protected]

References 1. Turner TT, Brown KJ. Spermatic cord torsion: loss of spermatogenesis despite return blood flow. Biol Reprod 1993; 49: 401-407. 2. Kristo BA, Palmer JS, Cromie WJ. Possible mechanisms for epididymal sparing during testicular ischemia. Urology 1999; 53: 435–439. 3. Ambrosio G, Flaherty JT, Duilio C. Oxygen radicals generated at reflow induce peroxidation of membrane lipids in reperfused hearts. J Clin Invest 1991; 87: 2056-2066. 4. Pieri C, Marra M, Moroni F. Melatonin: A peroxyl radical scavenger more potent than vitamin E. Life Sci 1994; 55: 272276. 5. Prillaman HM, Turner TT. Rescue of testicular function after acute experimental torsion. J Urol 1997; 157: 340-345. 6. Abasiyanik A, Dagdönderen L. Beneficial effects of melatonin compared with allopurinol in experimental testicular torsion. J Pediatr Surg 2004; 39: 1238-1241. 7. Akgur FM, Kilinc T, Aktug T, Olguner M. The effect of allopurinol pretreatment before detorting testicular torsion. J Urol 1994; 151: 1715-1717. www.smj.org.sa


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Free radical and antioxidant enzyme levels in VOC exposed workers

Free radical and antioxidant enzyme levels at exposure of volatile organic compounds in workers Sibel Bayil, Msc, Ahmet Celik, MD, Hulya Cicek MD, Iclal Geyikli, MD, Mehmet Tarakcioglu, PhD.

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arious effects of exposure to volatile organic compounds (VOCs) are currently a concern due to a widespread use of VOCs in industry. One occupation in which high exposure of organic solvents has been described is paint workers. Although the use of waterbased paint (WBP) has changed the exposure conditions, painters due to the lower emission of VOCs than with a solvent based paints (SBP), still carry the risk of exposure, since SBPs contain more reactive compounds such as benzene, toluene, biocide, and glycol ethers. In addition, these workers are exposed to all of the solvents due to the processing of factory’s manufacture schedule. The toxic effects of VOCs are caused by formation of reactive oxygen species (ROS). Free radicals are highly reactive species that is involved in cellular damage and can contribute to the conversion of normal cells. The ROS are believed to cause lipid peroxidation resulting in damage to biological membranes. Antioxidants such as enzymatic and nonenzymatic defense system are necessary to prevent cellular damage.1 Studies on painters include the association between exposure and health effects, yet little is known of biomarkers of these disorders. Also, the levels of malondialdehyde (MDA), superoxide dismutase (SOD), and total antioxidant capacity (TAC) parameters have not been studied in paint workers. Therefore, this study was to examine the effects of VOC on lipid peroxidation and antioxidant enzyme activities in paint workers. Twenty male paint workers were chosen as the exposure group and a similar number of healthy male, matched by age, were chosen as controls (Table 1). All painters were asked to answer a questionnaire that included questions regarding the age, smoking health, years worked as a paint worker, use of gloves, and personal respiratory protective equipment during working. The study was explained to the workers and approved by a local ethic commission. Venous blood samples obtained from the paint workers at the work site were codified and transported to the lab, where they were processed. Sera were separated and used without any treatment and erythrocytes were washed with 0.9% sodium chloride solution and centrifuged for 10 minutes. Washing was repeated 3 times. After a complete lysis, cellular particles were removed by centrifugation and the hemolysate was used for the determination of SOD activity. None of the paint workers in this study used gloves and protective 290


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respiratory equipment. They used SBP and WBP due to the factory’s schedule, and were therefore, exposed to a mixed type of solvent. All the paint workers had spent at least 5 years in the occupation. There were 9 smokers and 11 non-smokers in both the paint and control group (Table 1). The MDA concentration of paint workers was significantly higher than the control group (5.13 ± 0.72 nmol/mL versus 1.52 ± 0.45 nmol/mL, p0.05 >0.05 >0.05

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Data presented as mean ± SD of 20 subjects in each group

Free radical and antioxidant enzyme levels in VOC exposed workers

increase in SOD activity in erythrocytes in our study might have occurred to neutralize the excess superoxide anions originating from volatile organic compounds such as benzene, toluene, and thinner. Halifeoglu et al4 also found increased MDA and SOD activity in a study sample working with paint thinner.4 The antioxidant system have many components. The antioxidant capacity may give more relevant biological information compared to that obtained by the measurement of individual components, as it considers the cumulative effect of all antioxidants present in plasma and body fluids. We observed a significant decrease in the concentration of TAC in the paint group’s sera compared to control. These findings showed that the other antioxidant defense mechanism is used versus oxidative damage. A plausible explanation could be that the solvent exposure may cause inhibition of enzymes or depletion of substrate molecule (glutathione, GSH and so forth) and an increase in the concentration of ROS. Our data indicate that smoking is not a confounder for the association between solvent exposure and changes in blood parameters. Also, Pinto et al5 reported that smoking or alcohol intake did not correlate positively with the cytogenetic damage observed in outdoor painters. As a result, elevated MDA levels in the paint group may indicate that increased lipid peroxidation at exposure to long-term organic solvents. Whereas elevated SOD activity in the paint group compared to control, it is shown that the antioxidant system is activated against lipid peroxidation. Beside this, decreased TAC level in the paint group may indicate that another antioxidant system is used. Whichever the case is, our results permit us to conclude that paint workers studied represent a risk group and should be medically followed up with more frequent periodic examinations. These workers should take antioxidants and use gloves and protective equipment. Received 30th May 2006. Accepted 13th September 2006. From the Department of Biochemistry, Central Laboratory of Medical Faculty, Gaziantep University, Gaziantep, Turkey. Address correspondence and reprint requests to: Assist. Prof. Iclal Geyikli, Biochemistry Department of Medical Faculty, Vocational School of Health Gaziantep University, PO Box 39 Gaziantep University Campus, Gaziantep, Turkey. Tel. +9 (342) 3601200 / 2188. Fax. +9 (342) 3604423. E-mail: [email protected]

References 1. Gutteridge JM. Lipid peroxidation and antioxidants as biomarkers of tissue damage. Clin Chem 1995; 41: 1819-1828. 2. Riise T, Moen BE, Kyvik KR. Organic solvents and the risk of multiple sclerosis. Epidemiology 2002; 13: 718-720.

3. Karagözler AA, Mehmet N, Batcioglu K. Effects of long-term solvent exposure on blood cytokine levels and antioxidant enzyme activities in house painters. J Toxicol Environ Health A 2002; 65: 1237-1246. 4. Halifeoğlu İ, Canatan H, Üstündağ B, Inanç N. Effect of Thinner Inhalation on lipid peroxidation and some Antioxidant Enzymes of People Working with Paint Thinner. Cell Biochem Funct 2000; 18: 263-267. 5. Pinto D, Cebellos JM, Garcıa G, Güzman P, Del Razo LM, Vera E, et al. Increased cytogenetic damage in outdoor paints. Mutat Res 2000; 467: 105-111. ---------------------------------------------------------------------------------

Synchronous occurrence of chromosome-positive chronic leukemia and breast cancer

Philadelphia myelogenous

Ibrahim S. Tillawi, MBBCh, FASCP.

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he synchronous occurrence of non-treatment related Philadelphia chromosome positive (PhC+) chronic myelogenous leukemia (CML) and breast cancer (BC), is reported in this communication. The coincidental discovery of CML and BC is described in 2 patients, aged 37 and 39 years, who presented almost in a similar manner, where the diagnosis of chronic uncontrolled phase of CML was made upon working up the patient for radical surgery for a coexisting breast cancer. Both patients were treated with modified radical mastectomy with axillary clearance, which was followed by chemotherapy for both diseases. The first patient subsequently underwent autologous bone marrow transplantation, and is alive and well when last followed up nearly 27 months after the initial diagnosis. The second patient, who has a strong family history of breast cancer, died 18 months after initial diagnosis of uncontrolled metastatic breast cancer to the central nervous system. A 37-year-old Caucasian, gravida 3, para 3 and otherwise healthy female teacher, was admitted to work her up for a recently diagnosed BC by fine needle aspiration biopsy (FNAB). The mass (in the upper lateral quadrant) was painless and was initially felt while taking a shower and was found to progressively increase in size. The first FNAB carried out 4 weeks after breast mass discovery, was erroneously interpreted as a fibroadenoma. As the mass continued to enlarge, a second FNAB was carried and revealed the definitive presence of adenocarcinoma. Family history for BC was negative. She was scheduled for mastectomy, for which she underwent as part of her work up an admission,

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Synchronous occurrence of Philadelphia chromosome

complete blood count (CBC), which incidentally revealed the following incidental findings: hemoglobin 11.0 gm/dl, hematocrit 33%, white blood cell (WBC) 51,750, differential-lymphocytes 2%, segmented neutrophils 35%, bands 16%, metamyelocytes 5%, myelocytes 29%, promyelocytes 3%, blasts 2%, basophils 5%, eosinophils 1%, monocytes 2%, and platelets 374,000. Blood film examination was suggestive of a chronic uncontrolled phase of CML. Physical examination was remarkable (other than the breast mass) only for splenomegaly slightly below the level of the umbilicus. Subsequently, a bone marrow biopsy and aspiration were carried out, and the diagnosis of chronic myeloproliferative disorder most consistent with CML was made. Cytogenetic analysis revealed the presence of Philadelphia chromosome [46XX, t(9;22)(q34;q11.2)] in all of the 15 metaphases that were examined. Florescent in-situ hybridization (F.I.S.H.) for breakpoint cluster region - ABL was positive in 95% of the interphase cells for BCR/ABL translocation. The patient was then worked up by total body skeletal survey, bone isotope scans, mammography for the opposite breast, computed tomographic (CT) scans for the abdomen, pelvis and chest; patient was found to have a homogenously enlarged spleen and BC that was localized to one breast with only few regional lymph nodes enlargement. Modified radical mastectomy with axillary lymph node clearance confirmed the cancer state with 2 lymph node involvement by metastatic cancer. She was later placed on chemotherapy [Tamoxifen and Cyclophosphamide, methotrexate, and fluorouracil (C.A.F. regiment)] and hydroxyurea, for both diseases. Six months later, she underwent autologous bone marrow transplantation at the MD Anderson. She continued her follow-up visits with the local oncology team and continued to be disease free. She was last seen at follow up visit 27 months after the initial diagnosis, and was then disease free. A 39-year-old, gravida 4, para 4, Caucasian female housewife complaining of a retro-areolar hard painless fixed mass with associated nipple ulceration and inversion. The mass which was felt by palpation as “nodular in texture” showed progressive enlargement over the ensuing 4 months becoming almost confluent with associated nipple discharge thought to be related to breast feeding as she was lactating her baby. Subsequently, the patient underwent a FNAB of the mass, which revealed “suspicious for malignant cells”. She was scheduled for an excisional biopsy of the mass, during which her admission CBC revealed the following incidental findings: hemoglobin 11.0 gm/dl, hematocrit 33.0%, WBC 49,375, differentiallymphocytes 2%, segmented neutrophils 34%, bands 14%, metamyelocytes 4%, myelocytes 31%, 292


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promyelocytes 3%, blasts 2%, basophils 6%, eosinophils 4%, and platelets 610,000. Blood film examination was suggestive of a chronic uncontrolled phase of CML. A bone marrow biopsy and aspiration confirmed the presence of chronic myeloproliferative disorder, most consistent with CML. Splenic enlargement was documented by both physical examination and by abdominal ultrasound. Cytogenetic evaluation of the bone marrow cells, revealed the presence of Philadelphia chromosome [46XX, t(9;22)(q34;q11.2)] in all the 23 metaphases examined, and 88% of the interphase cells showing BCR/ABL translocation by F.I.S.H. She was worked up with total body and isotope bone scans, chest abdomen and pelvic CT scans, mammography for the opposite breast, and was found to have disease limited to the breast. A modified radical mastectomy with axillary lymph node clearance revealed multifocal infiltrating poorly differentiated ductal adenocarcinoma with metastasis to 23 out of 24 regional lymph nodes. She was subsequently placed on Tamoxifen and adjuvant chemotherapy (C.A.F.) in addition to hydroxyurea. Family history revealed a mother, and a sister who both died of breast cancer at an early age; she died after 18 months of uncontrolled breast cancer metastasis to the brain. Pathological findings. Bone marrow aspiration and biopsy in both patients showed similarly a cellularity of 90-95% (packed marrow), with myeloid predominance (hyperplasia), reticulin content was +1 to +2/+4, the myeloid: erythroid (M:E) ratio is 15-20:1, megakaryocytes are increased with many exhibiting single small nuclei; all myeloid stages of maturation are observed with expansion of the intermediate and late compartments, blasts did not exceed 5%. Coupled with the results of the cytogenetics, the overall picture was diagnostic of a chronic uncontrolled phase of CML. Keratin stains for the bone marrow biopsy samples were negative in both patients. The breast mass in the first patient measured 7.5 cms. Low power view of the tumor showed pushing margins and stellate central fibrosis entrapping foci of necrosis. High power examination showed an in situ comedo intraductal adenocarcinoma component constituted 25%, with the remaining tumor composed of infiltrating poorly differentiated ductal adenocarcinoma without any accompanying lymphovascular invasion. Both estrogen receptors (ER) and progesterone receptors (PR) were positive; with no p53 overexpression; no androgen receptor expression; Keratin (CK-7) positive; Her-2/neu is 0-1/3; and Ki67 score of 1/3; leukocyte common antigen (LCA) was negative; GCDFP-15 was positive. Two of the 18 regional lymph nodes show microscopic (subcapsular) metastasis. Margins of resection of the mastectomy sample were all free of tumor. The breast mass in the second patient was


composed of multiple confluent smaller masses that measure in maximum dimensions 5.5 cms. Sections were initially thought to represent granulocytic sarcoma (owing to the recent recognition of coexisting CML in the patient), but histopathologic examination of the mass revealed infiltrating poorly differentiated ductal adenocarcinoma, with widespread lymphovascular permeation. Examination of the mastectomy sample revealed the presence of multifocal adenocarcinoma in the other breast quadrants, and 23/24 regional lymph nodes show metastatic adenocarcinoma. She was ER and PR positive, her-2/neu 0-1/3. Keratin (CK-7) was positive; LCA was negative. Surgical margins of resection were tumor free. Improvements in the chemotherapy and radiotherapy regimens administered, or both, as well as immunosuppressive treatment following allogenic organ transplantation has increased the incidence of secondary hematological malignancies after therapy for primary neoplastic diseases as an important late complication of therapy in cancer patients. The occurrence of “Non-Treatment-related” leukemia on the other hand is exceedingly rare as a second primary and is reported to occur in older individuals (in Western countries).1 Treatment–related CML, which is indistinguishable from de novo CML and does not appear to have peculiar cytogenetics is a rare event, and its development seems to be associated with chemotherapy, radiotherapy, or immunotherapy.4 The term synchronously occurring cancers has been used to describe cancers diagnosed simultaneously in the same patient within a 6-month period between the diagnosis of the 2 cancers apart, whereas the term metachronous cancers is used to describe cancers occurring within a period of more than 6 months apart.2 In one study on 8428 autopsied cases, the average age of the examined cases was 64 years ± 2 years, the incidence of multiple primary tumors had an incidence of 0.8%, and 3.6 % of all cancer autopsied cases. First, primary malignant tumors were most common in the hematopoietic system and the cervix, second primary malignant tumors were most common in the lungs and the hematopoietic system.2 Age has been reported to play a significant role in multiple primary cancers in older individuals in the Western published data only, such a barrier against occurrence has not been confirmed from Middle-Eastern cases. Synchronous occurrence of primary epithelial neoplasm and a primary hematopoietic neoplasm is uncommon, most reported cases cite only the occurrence of BC after the use of chemotherapy for BC (namely, therapy–related). The only single report that clearly described an association between BC and chronic myelomonocytic leukemia (CMML) of the FAB classification, and

not Ph (+ve) CML, was that by Cavanna et al,3 who discovered the presence of the leukemia (CMML) 2 weeks after the diagnosis of operable breast cancer. Metachronous occurrence of such tumors; on the other hand, is more common where the second malignancy (usually hematopoietic neoplasm), develops due to the effects of chemotherapy, namely, secondary leukemia.4 Early metachronous is a related term used by some to refer to cancers occurring within a period of 3 years from initial diagnosis of the primary tumor. From the data by Specchia et al,1 the only epithelial neoplasms occurring simultaneously with de novo CML were those from the lung, prostate, bladder (one case each), and rectal cancer (2 cases); there has been no second malignancies with de novo CML cases reported from their series to arise from the breast. Oncologists know that the composition of chemotherapy, with particular reference to its high proportions of prednisolone, appears to enhance the leukemogenic effects of other chemotherapy agents used. The carcinogenic effects of adjuvant Tamoxifen treatment and radiotherapy for breast cancer are estimated to be in the range of 0.5%. Among 236 cases of hematological malignancies, synchronous solid cancers in the same patient were estimated to be in the range of 3%, none of their patients had BC. Others report a higher frequency of double cancer in the patients with hematopoietic neoplasms being close to 21.5% in patients of the older age group (>65 years), versus 2.6% in the patients of the younger age group.2,3 Although full karyotype was carried out for both patients, no other chromosomal abnormality was detected in both patients, other than that of Philadelphia chromosome. Ideally, a thorough search for specific gene abnormalities should have been undertaken, but that was not arranged at the time of patient management by the oncology team. Such synchronous de-novo malignancies most likely have to deal with genetic aberrations, although this was not substantiated in our study. Logically one presumes that BC related antigens might have been detected in these 2 patients, but as mentioned earlier these studies were not arranged for. Region of deletion on chromosome 22q13 as one of the reported allelic losses has been reported to occur in BC. Similarly, loss of the distal segment distal to M-bcr in a Philadelphia chromosome positive CML (or loss of heterozygosity at chromosome 9p(19), or allelic loss at 9p221-22, smallest common region in the vicinity of the CDKN2 gene in sporadic BC.5 Finally, one of the major cytological differential diagnoses in this setting of a poorly differentiated neoplasm in the presence of leukemia is granulocytic sarcoma, especially, if the sample is either qualitatively or quantitatively inadequate and if other difficulties existed in the sample www.smj.org.sa


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interpretation, due to the therapeutic implications, which are variable in both disease states. In conclusion, the simultaneous occurrence nontherapy related Philadelphia chromosome negative, CML, and BC in 2 female patients has been reported. The BC was identified earlier than the knowledge that simultaneously present CML was present in these patients, which was discovered incidentally upon working the patients up for breast cancer radical surgery. Received 5th June 2006. Accepted 30th August 2006. From the Department of Pathology, Division of Hematology, Riyadh Military Hospital, Kingdom of Saudi Arabia. Address correspondence and reprint requests to: Dr. Ibrahim S. Tillawi, Consultant in Hematopathology, Department of Pathology, Division of Hematology, Riyadh Military Hospital, Riyadh 11154, Kingdom of Saudi Arabia. E-mail: [email protected]

References 1. Specchia G, Buquicchio C, Albano F, Liso A, Pannunzio A, Mestice A, et al. Non-Treatment–related chronic myeloid leukemia as a second malignancy. Leuk Res 2004; 28: 115-119. 2. Lee TK, Myers RT, Scharyj M, Marshall RB. Multiple primary malignant tumors (MPMT): Study of 68 Autopsy cases (19631980). J Am Geriatr Soc 1982; 30: 744-753. 3. Cavanna L, Vallisa D, Di Stasi M, Fornari F, Buscarini E, Schena C, et al. Acute Myelocytic Leukemia and Chronic Myelomonocytic leukemia simultaneously with resectable Breast Cancer: A report of two Cases. Tumori 1992; 78: 356358. 4. Malacarne P, Bertusi M, Bariani L. Association of Breast Cancer and Chronic Myelogenous Leukemia. Riv Emoter Immunoematol 1977; 24: 128-134. 5. Iida A, Kurose K, Isobe R, Akiyama F, Sakamoto G, Yoshimoto M, et al. Mapping a new target region for allelic loss to a 2-cM interval at 22q113.1 in primary breast cancer. Genes Chromosomes Cancer 1998; 21: 108-112. ---------------------------------------------------------------------------------

Prevalence of risk factors of coronary heart disease among diabetic patients in Medina city Sami A. Al-Shammary, ABFM, JBFM.

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iabetes mellitus (DM) is a common, complex, serious, and costly disease. Nevertheless, it can affect any part or organ of the body, and coronary artery diseases are major contributors to morbidity. Thus, diabetic patients have a high incidence of silent myocardial infarction, and patients with diabetes type II have 2-3 fold more chances for atheroma related diseases.1,2 The overall prevalence of DM obtained from an epidemiological national study was 23.7% 294


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in Saudi Arabia. The prevalence in males and females was 26.2% and 21.5% (p10 year) patients (p=0.042). While compared FBS level, obesity class II showed high prevalence in the group with uncontrolled diabetes (7.8%) than the controlled (2%). Evidently, there are potential limitations of our study, however, we tried to reduce bias by random selection of primary health care units and patients’ files. However, even then it is warranted that selection of mini clinics produced selection bias due to patients who have diabetes with other severe complications come to the clinic, so the study has missed the bulk of diabetic patients. The sample size was also very small, and there was no control group to compare, again raising questions regarding internal and external validity. To conclude, the prevalence of risk factors of CHD is much higher among diabetic patients in this study,

Table 1 - Prevalence of risk factors of Coronary Heart Disease among diabetic patients as regard to gender, duration of disease and sugar level. Gender

Risk factors Male Total cholesterol Normal (10

Female

Fasting blood sugar level (mmol/L) ≤8.3

>8.3

66 32 58

(41.5) 29 (22.0) 20 (36.4) 24 0.6589

(39.7) (27.4) (32.9)

73 36 57

(43.9) 22 (21.6) 19 (34.3) 25 0.289

(33.3) (28.8) (37.9)

20 12 16

(41.7) 73 (25.0) 43 (33.3) 66 0.930

(40.1) (23.6) (36.3)

52 41 63

(33.3) 29 (25.8) 17 (39.6) 28 0.670

(39.1) (23.0) (37.8)

61 38 64

(37.4) 20 (23.3) 20 (39.3) 27 0.450

(29.9) (29.9) (40.2)

12 10 25

(25.5) 68 (21.3) 47 (53.2) 65 0.090

(37.8) (26.1) (36.1)

43 66 42 12 6

(25.4) 14 (39.0) 25 (24.8) 28 (7.1) 4 (3.55) 7 0.106* (20.0) 2 0.0005*

(17.9) (32.0) (35.9) (5.1) (8.9)

42 63 48 13 8

(19.4) (38.9) (30.5) (4.1) (6.9)

7 22 14 1 4

24

(14.8)

10

(14.5) 46 (45.8) 68 (29.9) 56 (2.0) 15 (8.0) 8 0.20 (19.2) 25 0.370*

(23.8) (35.2) (29.0) (7.8) (4.1)

(2.5)

(24.1) 14 (36.2) 28 (27.6) 22 (7.5) 3 (4.6) 5 0.710 (13.3) 11 0.942*

(39.7) 25 0.245*

(31.2)

58

(32.7) 35 0.042

(47.2)

24

(46.1) 69 0.180

(35.7)

34

68

(12.9)

*Yates correction

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Risk factors of CHD among diabetics in Medina city

and was found more prevalent in females as compared to males, except for smoking. Therefore, it is strongly recommended to implement a periodic screening program for high-risk patients, as well as the general population, and also start a public awareness program in order to prevent the risk factors at a primary level. Received 20th May 2006. Accepted 15th August 2006. From the Ministry of Health, Riyadh, Kingdom of Saudi Arabia. Address correspondence and reprint requests to: Dr. Sami A. Al-Shammary, Ministry of Health, PO Box 13534, Riyadh 11414, Kingdom of Saudi Arabia. Tel. +966 (1) 4961980. E-mail: [email protected]

References 1. Centers for disease control and prevention. Diabetes surveillance, 1999: the public health burden of diabetes mellitus in United States. 2. Webster MW, Scott RS. What cardiologists need to know about diabetes. Lancet 1997; 350: 523-527. 3. Al-Nozha MM, Al-Maatouq MA, Al-Mazrou YY, Al-Harthi SS, Arafah MR, Khalil MZ, et al. Diabetes mellitus in Saudi Arabia. Saudi Med J 2004; 25: 1603-1610. 4. Al-Humaidi MA. Probability of coronary artery disease among patients attending primary health care centers (PHCCs) in Southwest Saudi Arabia. Ethn Dis 2000; 10: 350-356. 5. Al-Turki YA. The prevalence of overweight and obesity among hypertensive and diabetic adult patients in a primary health care center. Saudi Med J 2000; 21: 340-343. 6. Garcia MJ, Mcnamara PM, Gordon T, Kannel WB. Morbidity and mortality in the Framingham population, sixteen years follow-up. Diabetes 1974; 23: 105-111. ---------------------------------------------------------------------------------

Light and scanning electron microscopic investigation of the changes in hair with Dyskeratosis congenita Hakan H. Celik, MD, PhD, Hakan Erbil, MD, Ilkan Tatar, MD, Mehmet B. Ozdemir, MD, PhD.

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yskeratosis congenita (DC), also called ZinsserCole-Engman syndrome, is a rare congenital syndrome characterized by atrophy and a reticular pigmentation of the skin, dystrophy of the nails and leukoplakia together with multisystem ectodermal and some mesodermal changes. The striking feature of the skin is the tan-gray mottled hyperpigmented or hypopigmented macules or reticulated patches that on some areas appear like a fine network. These lesions are located typically on the upper torso, neck, and face, although the extremities may also be involved. Other manifestations of the disease may be hyperhidrosis of 296


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the palms and soles, bullous conjunctivitis, gingival disorders, dysphagia resulting from esophageal strictures and diverticula, skeletal abnormalities, aplastic anemia, mental deficiency, and hipersplenism.1 Besides these manifestations, Yilmaz et al2 indicated that the diagnosis of DC was made with typical skin lesions, dystrophic toenails, thin and sparse hair, and neutropenia with decreased myelopoiesis in bone marrow. There have been many articles on DC in the literature, but there have been few ultrastructural studies characterizing it. However, the ultrastructural evaluation of the effects of DC in hair have not been shown yet. We aimed to investigate both light and scanning electron microscopy imaging of hair with DC. Hair specimens were obtained from 2 boys in different ages; one was 7 and the other was 9-year-old. Both children have showed typical clinical manifestations of DC. A total of 22 hair specimens picked from each individual have been examined. These samples were studied by light and scanning electron microscopes. For scanning electron microscopy, hairs were directly mounted on metal stubs, then sputtered with a 100 Å thick layer of gold in a Bio-Rad sputter apparatus. The specimens were examined with a JOEL SEM ASID 10 in 80 Kv.3 We evaluated hair specimens according to: shaft structure, cuticle pattern, filamentous-keratinized structures, and degeneration. Hair specimens taken from both patients had great similarities. In light microscopy, we examined thin and sparse hair with a longitudinal groove in the hair shaft and multiple patchy corruption of cuticle pattern (Figure 1a). In scanning electron microscopic examinations, we found some flattened areas, which disturbed the normal round shape of the hair shaft, and there were some cuticle irregularities in these regions and we also found, longitudinal groove or fissure like shaft changes with cuticular overlapping, the same as light microscopic findings (Figure 1b & 1c). Scanning electron microscopic examinations of the hair for different clinical syndromes could still be respected as new. Celik et al4 found destroyed cuticular pattern and degenerative areas on the surface of hair in Chédiak Higashi syndrome. They also found abnormal proliferation of cuticular cells, deformed hair surface structures in hair with hereditary trichodysplasia.5 Our study demonstrates the structural changes of the hair with DC in detail. Although our findings were not pathognomonic for DC, scanning electron microscopy might be used for distinctive diagnosis of DC. It can be used to differentiate hair with DC from other diseases effecting hair surface morphology. Scanning electron microscopy, is a multi-dimensional examination technique revealing easily comparable images. It is indispensable for diagnosis in various

SEM examination of hair with Dyskeratosis Congenita

a

c

b

Figure 1 - a) A light microscopic image showing thin and sparse hair with a longitudinal groove in hair shaft and multiple patchy corruption of cuticle pattern. b, c). Scanning electron microscopic images showing flattened areas, which disturb the normal round shape of hair shaft with some cuticle irregularities in these region. Also showing longitudinal groove or fissure like hair shaft changes with cuticular overlapping.

tissues as it permits considerable magnification. As it can be used in the Chédiak-Higashi syndrome4 and hereditary trichodysplasia,5 its routine usage in many dermatological hair diseases with surface alterations has resulted in valuable contributions to scientific literature. In the future, these unique scanning electron microscopic findings, that belong to the different multi-systemic syndromes, might be collected in an atlas for distinctive diagnosis of the diseases. Acknowledgment. The study was supported by a research grant from the Scientific Research Office of Hacettepe University, Ankara, Turkey. Received 26th March 2006. Accepted 16th August 2006. From the Department of Anatomy (Celik, Tatar), Hacettepe University, Faculty of Medicine, Department of Dermatology (Erbil), Gulhane Military Medical Academy, Ankara, and the Department of Anatomy (Ozdemir), Pamukkale University, Faculty of Medicine, Denizli, Turkey. Address correspondence and reprint requests to: Dr. Ilkan Tatar, Department of Anatomy, Hacettepe University, Faculty of Medicine, 06100 Ankara, Turkey. Tel. +90 (312) 3052101. Fax. +90 (312) 3107169. E-mail: [email protected]

References 1. Fitzpatrick TB, Eisen AZ, Wolff K, Freedberg IM, Frank Austen K, editors. Dermatology in General Medicine, 5th ed. New York (NY): McGraw-Hill; 1999. p. 729-751. 2. Yilmaz K, Inaloz HS, Unal B, Guler E. Dyskeratosis Congenita with isolated neutropenia and granulocyte colony-stimulating factor treatment. Int J Dermatol 2002; 41: 170-172. 3. Celik HH, Aldur MM, Ozdemir B, Ilgi S. The usage of scanning electron microscope in microscopic anatomy. J Morphol 2001; 9: 39-42. 4. Celik HH, Tore H, Balci S, Bagcicek O, Ozdemir B. Scanning electron microscopic examination of the hair in ChédiakHigashi syndrome. Case Rep Clin Pract Rev 2002; 4: 252-255. 5. Celik HH, Surucu SH, Aldur MM, Ozdemir BM, Karaduman AA, Cumhur MM. Light and scanning electron microscopic examination of late changes in hair with hereditary trichodysplasia (Marie Unna hypotrichosis). Saudi Med J 2004; 25: 1648-1651.

Fundamental rights of infants are guaranteed in Islam - Breastfeeding is mandatory Iftikhar A. Khattak, BSc Hons, MSc Hons, PhD, Niamat Ullah, BSc Hons, MSc Hons.

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he Qur-an advocates breastfeeding as the recommended method of nourishing infants. There are several verses in the Holy Book, which put stress on the practice of breastfeeding. Every Muslim mother has to feed her baby with her own milk for the first 2 years of the baby’s life. Some schools of thoughts from the Muslim world consider it mandatory for a mother to feed her milk for at least 2 years. Breastfeeding has many advantages as proven by the modern day science. Breastfeeding is not only beneficial to the child but also it is good for the mother’s own health. It helps the mother to return to her pre-pregnant size and weight. It helps the uterus to involute under the effect of oxytocin secreted by the pituitary gland and released by reflexes through suckling. Breastfeeding protects the baby from diarrhea, chest infections, respiratory, and gastrointestinal diseases. Those who do catch the diseases are less morbid and have a much lower mortality rate than those who are bottle-fed. There has been a debate as for how long breastfeeding be continued. Unfortunately, there is no universally accepted position that clarifies the optimal length of breastfeeding. As far as the Qur-an is concerned, it is 2 years. This fact has been supported by World Health Organization (WHO) and United Nations Children’s Fund (UNICEF), and investigated extensively and supported widely by www.smj.org.sa


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Fundamental rights of infants to breastfeeding guaranteed in Islam

other organizations. It is imperative that the medical community supports the research that shows extended breastfeeding as beneficial to the baby in the long term. Review of the scientific facts. Benefits of milk. Human milk has been shown to be beneficial to the infants, mothers, and the whole family as indicated in some studies conducted over the years. Some advantages from these include health, nutritional, immunological, developmental, psychological, social, economic, and environmental benefits. The policy statement entitled “Breastfeeding and the Use of Human Milk”, published by the American Academy of Pediatrics (AAP) in 1997 emphasized on breastfeeding as the most effective tool for early infant development. The recommendations, as a result, are consistent with the goals and objectives of Healthy People 2010, which strongly stresses breastfeeding to infants. From a child health point of view, human milk is species-specific making it uniquely superior for infant feeding. Premature children get enormous benefits of host protection and improved developmental outcomes compared with formula-fed premature infants, despite the efforts of formula manufacturers, man-made formula remains significantly different from breast milk.1 Infections of children and breastfeeding. Studies2 on infection and breastfeeding have shown that feeding infants with human milk decreases the incidence and or severity of a wide range of infectious diseases including bacterial meningitis, diarrhea, respiratory tract infection, necrotizing enterocolitis, otitis media, urinary tract infection, and late-onset sepsis in preterm infants. In addition, breastfeeding has been shown to reduce post-neonatal infant mortality rates by 21%. The immune factors in milk provide better protection to babies. Decreased leukemia, improved cognitive achievements, decrease in rates of sudden infant death syndrome, and reduction in incidence of insulin-dependent (type-1) and non–insulin-dependent (type-2) diabetes mellitus have been shown the positive outcomes of breast feeding. Also, lymphoma, overweight and obesity, hypercholesterolemia, and asthma in older children have been shown to be closely related with prolonged breastfeeding. Benefits to mothers. Breastfeeding is good for the mothers’ own health. The maternal benefits include decreased postpartum bleeding and more rapid uterine involution attributable to increased concentrations of oxytocin, decreased menstrual blood loss and increased child spacing attributable to lactational amenorrhea, earlier return to pre-pregnancy weight, decreased risk of breast cancer, decreased risk of ovarian cancer, and possibly decreased risk of hip fractures and osteoporosis 298


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in the postmenopausal period. In China, women who breastfed for 2 years or more reduced their risk of breast cancer by 50%.2 Benefits to the community. From the viewpoint of the community, breastfeeding has been of enormous national savings. Decreased parental employee absenteeism and associated loss of family income, more time for attention to siblings and other family matters as a result of decreased infant illness, decreased environmental burden for disposal of formula cans and bottles, and decreased energy demands for production and transport of artificial feeding products.2 Duration of breast feeding. There has been a debate in the health circles on duration of breastfeeding. In a survey by the AAP conducted in 2000, approximately 25% of the participants favored breastfeeding up to 2 years.3 The WHO and UNICEF stated that breastfeeding should be continued for the first 2 years of life.4 Many studies support the WHO’s recommendations. The research on breastfeeding documents the advantages of extended breastfeeding to the general health, growth and development of infants at every socio-economic level, while the optimal duration of breastfeeding continues to be discussed and investigated throughout the world. Finally, recognizing that the scientific evidence indicates that breastfeeding is one of the most important contributors to infant health, the United States has reached the conclusion to increase breastfeeding. The new recommendations in the National Health Observations for 2010 include goals of 75% of mothers initiating breastfeeding, 50% of infants still receiving breast milk at 6 months, and 25% of infants still breastfed at 12 months.1 Qur-anic Instructions. It is stated in the Qur-an: “The mothers shall give suck to their offspring for two whole years, for him (the father) who desires to complete the term. But he (the father) shall bear the cost of their food and clothing on equitable terms. No soul shall have a burden laid on it greater than it can bear. No mother shall be treated unfairly on account of her child. No father, on account of his child, and here shall be chargeable in the same way. If they both decide on weaning, by mutual consent, and after the consultation, there is no blame on them, if ye decide on a foster-mother for your offspring there is no blame on you, provided ye pay the foster mother what ye offered, on equitable terms” (2:234). These are the regulations in case of divorce. The rights of the child, however, are safe guarded. The principle of care of the child applies equally to the father and the mother in wedlock: each must fulfill his or her part in fostering of the child. By mutual consent they can agree to some course that is reasonable and equitable, both as regard the period before weaning (the maximum being 2 years) and the engagement of a wet-nurse, or (by analogy) for artificial feeding. This is

Fundamental rights of infants to breastfeeding guaranteed in Islam

what is telling us that breastfeeding is to be continued for 2 years completely, even if the parents are divorced. However, the father of the child shall bear the cost of the mother’s food and clothing on a reasonable basis. And if it is decided to hire a foster mother to suckle the child, the father will bear all the costs of the foster mother. At another place, it is quoted; “His mother bore him by strain after strain. His nursing (suckling) period lasts two years. Be grateful to Me and to your parents” (31:14). The set of milk teeth in a human child is completed at the age of 2 years, which is therefore the natural extreme of breastfeeding. At another place we read in the Holy Qur-an; “We have enjoined on man, kindness to his parents: in pain did his mother bear him, and in pain did she gave him birth, the carrying of the child to his weaning is (a period of ) thirty months” (46:15). The maximum period of breastfeeding (2 years) is again in accordance with the time that the first dentition is ordinarily completed in a human child. The lower milk incisors in the center come out between the sixth and ninth months; then the milk teeth come out at intervals, until canines appear. The second molars come out at approximately 24 months, and with them the child has a complete apparatus of milk teeth. Nature now expects him to chew and masticate and be independent of his mother’s milk completely. On the other hand, it hurts the mother to feed from the breast after the child has a complete set of milk teeth. From these verses, we can understand that mothers are encouraged to nurse their babies for a period of 24 months. Even if the mother is divorced, she should be supported in her nursing of her child for that period. The father should pay her cost of living, as she will be fully occupied by nursing his offspring. If the parents are separated or divorced they should frequently counsel each other for the sake of the child’s future. If it is decided that the mother cannot nurse the baby (for example, if she lacks milk in her breasts), then a wet nurse must be asked to help. The wet nurse in Islam is a foster mother; she is also considered as the child’s mother. Her children become the baby’s brothers and sisters, and her husband becomes another father for the baby. If the mother dies, the baby’s heir should support both the nursing mother and her baby for the period of breastfeeding. The custody of the baby remains in the hands of the nursing mother. Received 25th February 2006. Accepted 16th August 2006. From the Department of Human Nutrition, Agriculture University, Peshawar, Pakistan. Address correspondence and reprint requests to: Dr. Iftikhar A. Khattak, Lecturer and Junior Research Specialist, Department of Human Nutrition, Agriculture University, Peshawar, NWFP Pakistan. Tel. +92 (91) 9216570 Ext. 3165. Fax. +92 (91) 9216520. E-mail: [email protected]

References 1. Healthy People 2010: Objective: Maternal, infant and child health. Available on internet. http://www.health.gov/ healthypeople/Document/html/volumse2/16MICH:36-37. Accessed on 16/1/2006. 2. American Academy of Pediatrics. Policy statement. Breastfeeding and the use of human milk. Pediatrics 2005; 115: 496-506. 3. American Academy of Pediatrics: Summary Report ACQUIP Exercise 3: Breastfeeding promotion and the healthy newborn infant. 2000. 4. Michels KB, Trichopoulos D, Rosner BA, Hunter DJ. Being breastfed in infancy and breast cancer incidence in adult life: results from the two nurses’ health studies. Am J Epidemiol 2001; 153: 275. ---------------------------------------------------------------------------------

Protozoa infection and its associated factors among urban babies 6 to 24 months age, in Semnan, Iran Raheb Ghorbani, PhD, Seyed M. Sadat-Hashemi, PhD, Ramin Pazooki, PhD.

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arasitic infection is one of the most important health problems in the entire world, hence, there was no region in the world that free from parasites. Some of them can induce anemia, malnutrition, diarrhea, and other infections. Parasitic infection depends on geographical area and time, demographic factors, health, knowledge and attitudes, socioeconomic conditions, and others; hence, it has a wide range. The high risk category are the children. The range of parasite infections can be wide, due to the variety of the weather and the socio-cultural conditions in Iran. In our research, we surveyed the status of parasites between 6-24 months in Semnan, Iran. The surveyed zones in this study were the urban areas of Semnan province nearly in the central north of Iran, which has been divided into 2 regions of wet-cold and dry-warm weather. A total of 482 children under 2 years old were surveyed (399 in warm and 83 in cold regions). The mothers that were referred to the health center in Semnan province and had children under 2 years, were given the sample container containing 30 cc of Phenol Alcohol Formaldehyde (PAF), an applicator for sampling accompanied by a questionnaire for filling up. They were trained to pour the stool of the children into the container with the help of the applicator, and shake it until the stool becomes homogenous. The samples were examined by direct and formalin-ether methods. For staining of the samples, Thionin was used, which is the preferred color for the samples fixed www.smj.org.sa


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Protozoa infection and its associated factors among urban babies Table 1 - Prevalence (%) of protozoa’s Infections in children.

Weather Warm Cold Gender Boy Girl Age (months) 6-11 12-18 19-24 Birth order 1 2 3 >4 Family size 3 4 5 >6

Dientamoeba fragilis

Endolimax nana

Entamoeba histolytica

Entamoeba hartmanni

Chilomastix mesnili troph

Blastocystis hominis

Giardia lamblia

Sample size n=482

Factor

Entamoeba coli

Protozoa’s name Total infection

P-value

0.137

(11.1) (13.2)

0.478

(0) (0.7) (0)

(3.6) (14.9) (24)

35 years and due to more experience in baby caring.

One of the most important results of our survey is a significant relationship between the type of the child’s consumed milk and the infection, those children who consumed cow’s milk or other foods were more infected in comparison with the children who were fed with their mothers’ or formula milks. Since there is piped drinking water for all urban population in Semnan, and there is a permanent control system foe testing the quality and safety of drinking water, therefore, we conclude that drinking water is not a source for infection. From this research, Giardia lamblia was the most frequent parasite in Semnan, which can be one cause of growth delay and decrement of the children’s weight. To prevent the parasites and the problems due to them, the dynamic education for mothers is a requirement. Acknowledgment. The authors would like to thank Abolfazl Amozadeh and Omran Asadi and all the colleagues for their help in this research, and with the support of the research deputy of Semnan University of Medical Sciences. Received 21st December 2005. Accepted 31st July 2006. From the Department of Social Medicine (Ghorbani, Sadat-Hashemi) and the Department of Parasitology (Pazooki), Faculty of Medicine, Semnan University of Medical Sciences, Semnan, Iran. Address correspondence and reprint requests to: Dr. Raheb Ghorbani, Department of Social Medicine, Faculty of Medicine, Semnan University of Medical Sciences, 5th Km Damghan Road, Semnan, Iran. Tel. +98 (231) 4441420. Fax. +98 (231) 4440225. E-mail: [email protected]

References 1. Cifuentes E, Suarez L, Espinosa M, Juarez-Figueroa L, MartinezPalomo A. Risk of Giardia intestinalis infection in children from an artificially recharged groundwater area in Mexico city. Am J Trop Med Hyg 2004; 71: 65-70. 2. Arthurs S, Heinz KM, Thompson S, Krauter PC. Effect of temperature on infections, development and reproduction of the parasitic nematode Thripinema nicklewoodi in Frankliniella occidentalis. BioControl 2003; 48: 417-429. 3. Park SK, Kim DH, Deung YK. Status of intestinal parasite infections among children in Bat Dambang, Cambodia. Korean J Parasitol 2004; 42: 201-203. 4. Moura EC, Bragazza LM, Coelho MF, Aun SM. Prevalence of intestinal parasitosis in schoolchildren. J Pediatr (Rio J) 1997; 73: 406-410. 5. Saksirisampant W, Nuchprayoon S, Wiwanitkit V, Yenthakam S, Ampavasiri A. Intestinal parasitic infestations among children in an orphanage in Pathum Thani province. J Med Assoc Thai 2003; 86: S263-S270.

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