Natural adjuvants (PC and G2) induce activated natural killer cells with NKG2D expression and cytotoxic properties in colorectal cancer.

Aim: This study aimed to investigate the effects of natural adjuvants (G2 and PC) to activate natural killer cells in colorectal cancer. Background: Natural killer (NK) cells are an element of the innate immune system that can recognize and kill cancer cells and provide hope for cancer therapy. One of the current methods in cancer immunotherapy is NK cell therapy. Immunotherapy with NK cells has been limited because of the low number and cytotoxicity level of NK cells. Natural adjuvants such as PC and G2 may stimulate the immune system. It seems that these adjuvants could increase cytotoxic NK cells. Methods: Twelve patients with colorectal cancer and six healthy individuals qualified for inclusion in this study. Peripheral blood mononuclear cells (PBMCs) from each patient with two distinctive concentrations (105and 5×104 cells/well) were treated with Interleukin2 (IL2), PC, and G2 adjuvant separately. The NK cell's surface markers, including CD16, CD56, and NKG2D, were evaluated by flow cytometry. The cytotoxicity effect of treated PBMCs as effector cells against NK sensitive cell line (K562) was assessed using the LDH assay method. Results: The results revealed a significant increase in the level of CD16+NKG2D+ NK cells in PBMCs treated with the G2 group compared with the control group in CRC PBMC (p<0.001) as well as the normal PBMC group (p < 0.01). In addition, the results indicated a significant increase in the level of CD56+NKG2D+ cells in the PBMC treated with PC (p < 0.05) and G2 (p < 0.001) groups compared with the PBMC group. The cytotoxicity result of PBMC from CRC patients in 10:1 ratio of the effector: target showed that the cells' cytotoxicity in the PBMCs treated with PC (p<0.01) and G2 (p<0.05) was significantly higher than the untreated PBMC. Conclusion: According to the result of this study, it can be stated that the PC and G2 adjuvants could be candidates for inducing cytotoxic natural killer cells.

Seroprevalence and molecular evaluation of toxoplasmosis in children with cancer in Khuzestan province, Southwest of Iran.

Toxoplasma gondii is an intracellular parasite with global distribution. Toxoplasmosis in individuals with normal immune system is usually asymptomatic, but in immunocompromised patients may lead to death if not cured. In this study, the prevalence rate of acute and chronic toxoplasmosis in children with cancer was investigated using serological and molecular methods. Blood samples were taken from 372 children with cancer in Shafa hospital in Ahvaz, southwest of Iran. Anti-T. gondii IgG and IgM antibodies were investigated by ELISA. The presence of Toxoplasma in the blood samples was evaluated by Nested PCR. Among 372 children with cancer, 155 (41.7%) were positive for anti-T. gondii IgG antibodies and 24 (6.4%) were positive for anti-T. gondii IgM antibodies, as well. In IgG avidity test, 34 (22%) had antibodies indicating acute phase and 121 (78%) had antibodies indicating chronic phase. The Nested PCR results were showed T. gondii parasite in 34 (100%) patients among 34 IgG antibody-positive patients with acute infection, among 16 IgG antibody-positive patients with chronic infection, 10 patients were indicative of T. gondii and 6 patients were not indicative of T. gondii. A total of 50 cases, 44 (88%) were T. gondii-positive and 6 (12%) were T. gondii-negative in Nested PCR. This study showed high prevalence of toxoplasmosis in children with cancer. Results of serological techniques (ELISA and IgG avidity) had a higher overlap with Nested PCR in identifying T. gondii of seropositive patients.

The first Molecular genetics analysis of individuals suffering from nephropatic cystinosis in the Southwestern Iran

Objective: Nephropatic Cystinosis (NC) is a rare metabolic disorder due to mutation in the CTNS gene in which more than 90 different mutations have already been reported so far. This study was performed to investigate mutations of the CTNS gene and its promoter in a number of Iranian patients with NC. Methods: Polymerase chain reaction and direct sequencing were performed for molecular characterization of the CTNS gene in 25 patients from 24 unrelated Iranian families with NC. Results: None of the patients showed the 57kb deletion in heterozygous or homozygous manner. One was homozygous for a novel mutation, which was termed as "c.153-155insCT", while one of the cases was homozygous and another was compound heterozygous for the second novel mutation c.923G>A. Moreover three known mutations c.18-21delGACT, c.1017G>A, and c.681G>A in 11 of the patients were detected. No apparent mutation was observed in the rest of patients (44%, n=11). Conclusion: The present data exhibit a fundament for molecular carrier detection and prenatal diagnosis of a relatively large percentage of Iranian patients suffering from NC, at least in the Southwestern Iran, where Arab ethnicity is one of the common ethnicities of the region (AU)

Objetivos: La cistinosis nefropática (CN) es una rara enfermedad metabólica que se debe a la mutación del gen CNTS, para que el que ya se han registrado más de 90 mutaciones diferentes. El estudio se realizó para investigar las mutaciones del gen CTNS y su promotor en un determinado número de pacientes iraníes con CN. Métodos: Para la determinación molecular del gen CTNS en 25 pacientes con CN procedentes de 24 familias iraníes no emparentadas, se han llevado a cabo análisis con reacción en cadena de la polimerasa y con secuenciación directa. Resultados: Ninguno de los pacientes mostró deleción de 57-kb ni heterocigócita ni homocigótica. Uno resultó homocigoto para una mutación no descrita que se denominó «c.153-155insCT», mientras que otro de los casos resultó homocigoto y otro heterocigoto compuesto para una segunda mutación no descrita, c.923G>A. Además, se detectaron otras tres mutaciones conocidas c.18-21delGACT, c.1017G>A y c.681G>A en 11 pacientes. En el resto de los pacientes (44 %, n = 11) no se observaron mutaciones aparentes. Conclusión: Los datos de este estudio muestran un fundamento para la detección de transportadores moleculares y el diagnóstico prenatal de un porcentaje relativamente elevado de pacientes iraníes que sufren CN, al menos en el sudoeste de Irán, donde la etnia árabe es una de las comunes en la región (AU)

The first molecular genetics analysis of individuals suffering from nephropatic cystinosis in the Southwestern Iran.

OBJECTIVE: Nephropatic Cystinosis (NC) is a rare metabolic disorder due to mutation in the CTNS gene in which more than 90 different mutations have already been reported so far. This study was performed to investigate mutations of the CTNS gene and its promoter in a number of Iranian patients with NC. METHODS: Polymerase chain reaction and direct sequencing were performed for molecular characterization of the CTNS gene in 25 patients from 24 unrelated Iranian families with NC. RESULTS: None of the patients showed the 57 kb deletion in heterozygous or homozygous manner. One was homozygous for a novel mutation, which was termed as "c.153-155insCT", while one of the cases was homozygous and another was compound heterozygous for the second novel mutation c.923G>A. Moreover three known mutations c.18-21delGACT, c.1017G>A, and c.681G>A in 11 of the patients were detected. No apparent mutation was observed in the rest of patients (44%, n=11). CONCLUSION: The present data exhibit a fundament for molecular carrier detection and prenatal diagnosis of a relatively large percentage of Iranian patients suffering from NC, at least in the Southwestern Iran, where Arab ethnicity is one of the common ethnicities of the region.

Cerebellar neuroblastoma in 2.5 years old child.

Neuroblastoma is the third most common malignancy of childhood, after leukemia and brain tumors. Only 2% of all neuroblastoma occur in the brain. Primary cerebellar neuroblastoma is an specific subset of Primitive Neuroectodermal Tumors (PNET). Meduloblastoma is a relatively common and well-established entity, consisting of primitive and multipotential cells that may exhibit some evidence of neuroblastic or gliad differentiation. But cerebellar neuroblastoma with ultrastractural evidence of significant neuroblastic differentiation is extremely rare. We report a rare case of neuroblastoma in the cerebellum. A 2.5-year-old Iranian boy presented with vomiting and nausea in the morning and ataxia. CT scan showed a tumor mass in the cerebellum and the report of radiologist was medulloblastoma. Light microscopic assay showed a small cell neoplasm with lobules of densely packed cells (lobulated pattern) and better differentiated cells. Neuron-Specific Enolase was positive. Pathologic diagnosis confirmed the existence of cerebellar neuroblastoma. Chemotherapy followed surgical removal. No relapse occurred 12 months after surgery.

Clinical characteristics and the prognosis of childhood rhabdomyosarcoma in 60 patients treated at a single institute.

Rhabdomyosarcoma (RMS) is the most frequent soft tissue sarcoma in children. The aim of study was to retrospectively review the treatment results of childhood rhabdomyosarcoma and identify prognostic factors. 60 children with rhabdomyosarcoma treated between 1996 and 2002 in Shafa Hospital were reviewed. The data were analyzed for clinico-epidemiological factors. Age, gender, race, histology type, primary site, tumor size and intergroup rhabdomyosarcoma study (IRS) group were evaluated. The primary site of involvement was orbit in 6 cases (10%) head and neck nonparameningial in 12 cases (20%), parameningial region in 12 cases (20%). The histological findings were as follows: 12 cases (72.5%) for embryonal, 6 cases (10%) for alveolar and 11 cases (17. 5%) for botryoid type. With respect to the IRS III (15%) were group II, 32 (52.5%) were group III and 24 cases (40%) were group IV. The 5-year survival rate was 47.9%. Primary tumor site (P=0.0003), and histology (P=0.05) were associated significantly with survival after recurrence. Among the variables, age, gender, regional lymph node involvement, and IRS group did not affect 5-year survival but the type and time of recurrence (P=0.0002), and its relation with therapy (P=0.0001) were associated with survival. This study showed that overall survival for rhabdomyosarcoma is dependent on histological subtype, primary site, disease group, duration of disease before treatment. The outcome for infant with RMS is less satisfactory than older children and the patients aged 1-9 years had the best 5 year survival.

Identification of a novel de novo mutation in the NIPBL gene in an Iranian patient with Cornelia de Lange syndrome: A case report.

BACKGROUND: Cornelia de Lange syndrome is characterized by dysmorphic facial features, hirsutism, severe growth and developmental delay. Germline mutations in the NIPBL gene with an autosomal dominant pattern and in the SMC1A gene with an X-linked pattern have been identified in Cornelia de Lange syndrome. CASE PRESENTATION: A two-month-old Iranian boy who showed multiple congenital anomalies was referred to the genetic center of a welfare organization in southwest Iran. He was the second child of a non-consanguineous marriage, born after full term with normal delivery. His birth weight was 3110 g, his length was 46 cm and his head circumference was 30 cm. Both parents were clinically asymptomatic, with no positive history of any deformity in their respective families. CONCLUSIONS: Sequencing of the NIPBL gene from our patient revealed a single-base deletion of thymidine in exon 10 (c.516delT). This mutation presumably results in premature termination at codon 526. We did not observe this mutation in the parents of our patient with Cornelia de Lange syndrome. The results presented here enlarge the spectrum of NIPBL gene mutations associated with Cornelia de Lange syndrome by identifying a novel de novo mutation in an Iranian patient with Cornelia de Lange syndrome and further support the hypothesis that NIPBL mutations are disease-causing mutations leading to Cornelia de Lange syndrome.

Renal tubular dysfunction in pediatric patients with beta-thalassemia major.

To evaluate the prevalence of renal tubular dysfunction in children with ß-thalassemia (ß-T) major, we studied the glomerular and tubular function in 140 children with ß-T major and compared them to a healthy control group at our center from May 2007 to April 2008. Fresh first morning samples were collected from each patient and analyzed for sodium, potassium, calcium (Ca), protein, uric acid (UA), creatinine (Cr), urine osmolality and urinary N-acetyl-ß-D-glucosaminidase (UNAG) activity. Blood samples were also collected for complete blood count, blood urea nitrogen (BUN), fasting blood sugar, serum creatinine (SCr), electrolytes, and ferritin before transfusion. Among the study patients, 72 were males, and the mean age was 11.5 (ranging 7-16) years. SCr levels were all within normal limits and all of them had normal glomerular filtration rate (GFR). The mean UNAG was 17.8 IU/L in the study patients (normal 0.15-11.5 IU/L) and 3.2 IU/L in the control group (P < 0.001). Of the 82 study patients who had elevated level of UNAG, 58 (62.4%) had high blood levels of ferritin also (r = 0.2, P < 0.001) and 13 (15.9%) patients had hypercalciuria also (UCa/UCr > 0.21) (P = 0.006). Nine (6.4%) thalassemic patients with a mean age of 12 years had proteinuria (Upr/UCr > 0.2). Sixty-nine (49.3%) out of the 140 patients and 45 (65.2%) of the patients having UNAG had uricosuria also (UUA/UCr > 0.26). Ten (7%) patients had microscopic hematuria and 10 (7%) patients with a mean age of 13.5 years had glucosuria or diabetes mellitus. We conclude that tubular dysfunction is a relative common complication of the ß-T major; UNAG and its index are the best to detect renal tubular dysfunction in these patients. Currently, periodic measurement of UCa/UCr and UUA/UCr ratios as well as urinalysis are recommended.

Comprehensive spectrum of the ß-Thalassemia mutations in Khuzestan, southwest Iran.

ß-Thalassemia (ß-thal) is characterized by reduction or absence of ß-globin gene expression. We describe the spectrum of mutations observed in a large cohort of ß-thal carriers in Khuzestan, Southwest Iran. All together 1,241 blood samples from individuals with decreased mean corpuscular volume (MCV) and elevated Hb A(2) levels, were analyzed either by reverse dot-blot or by direct sequencing of the HBB gene. We found 42 different mutations associated with ß-thal and identified eight common ß-globin variants, namely, Hb S [ß6(A3)Glu→Val], Hb C [ß6(A3)Glu→Lys], Hb D-Punjab [ß121(GH4)Glu→Gln] and Hb O-Arab [ß121(GH4)Glu→Lys]. No mutations were found in two individuals. The distribution is characteristic of a heterogenous population with three preferential mutations being present [codons 36/37 (-T), IVS-II-1 (G>A) and IVS-I-110 (G>A)] at a frequency of 20.5, 20.0 and 14.2%, respectively, followed by 39 mutations in decreasing frequencies from 5.2 down to 0.1%. These data are of importance when planning prevention strategies in the country.

A novel COL7A1 gene mutation in an Iranian individual suffering dystrophic epidermolysis bullosa.

Dystrophic epidermolysis bullosa is a heritable skin disorder with dominant and recessive genetic patterns. Numerous studies underline that both forms are caused by mutations of the COL7A1 gene, which encodes collagen type VII. It has been reported that most mutations detected in the recessive disease form are nonsense mutations or small insertions or deletions leading to frameshift and premature translational termination, which tend to produce severe phenotypes. In contrast, missense mutations causing amino acid substitutions, which result in variable phenotypes, predominate in the dominant form of dystrophic epidermolysis bullosa. Genomic DNA from the patient and parents was subjected to PCR amplification of the coding region of the COL7A1 gene. Direct sequencing of the PCR products revealed a homozygous single-base deletion in the patient (c.6269-6270delC). The parents were heterozygous for the same mutation. This deletion is a novel mutation in the human COL7A1 gene based on comparisons with the Human Genome Mutation Database. To our knowledge, this is the first report of dystrophic epidermolysis bullosa in an Iranian patient confirmed by molecular diagnosis.

alpha-thalassemia mutations in Khuzestan Province, Southwest Iran.

Although alpha-thalassemia (alpha-thal) is the most common hereditary hemoglobin (Hb) disorder in Iran, no comprehensive data are so far available on the prevalence of the disease in the province of Khuzestan in Southwest Iran. This study investigates the spectrum of alpha-thal mutations in this region. One hundred and twenty-one subjects from Khuzestan Province, Iran, were initially tested for the three most common Iranian alpha-thal mutations (- alpha3.7, -alpha4.2, and --MED) by gap-polymerase chain reaction (gap-PCR). Reverse hybridization test strips and DNA sequencing were used to identify additional alpha-globin mutations. A total of 131 mutated alpha-globin alleles were identified in these patients. Of the 13 mutations that were detected in Khuzestan Province, Iran, the - alpha3.7 single gene deletion was the most frequently identified variant, representing 62.6% of the total; we also observed significant numbers of individuals with compound heterozygous mutations. On the basis of our results, we strongly recommend screening for the most common mutations to improve the molecular diagnosis of anemia in this region.

Una breve Comunicación desde Irán sobre el consumo de tabaco entre los estudiantes de Medicina, los Médicos Residentes y los Médicos Adjuntos

Objetivo : Evaluar la prevalencia de fumadores entre los estudiantes de Medicina iraníes, Médicos Residentes y Adjuntos. Método: Los sujetos (532 varones y mujeres) se seleccionaron al azar en la Unversidad de Shiraz de Ciencias Médicas y a todos ellos se les pasó una encuesta. Resultados: De acuerdo con los resultados obtenidos un 16,79 per cent de varones y 0,69 per cent de mujeres eran fumadores. Entre los estudiantes de Medicina, encontramos que el 18,48 per cent.de los estudiantes de primer año, el 12,5 per cent de los de segundo año, el 7,l9 per cent de tercer año y el 16,95 per cent de los estudiantes internos eran fumadores. En cambio, entre los Médicos Residentes sólo el 11,11 per cent eran fumadores y el 7,57 per cent de los Médicos Adjuntos fumaban. Las causas más habituales para fumar eran la necesidad de evitar los síntomas tras la retirada, el placer que supone, y por último como forma de eliminar la tensión. El tabaco con filtro era el que más se consumía. La media de cigarrillos por día era de 6,29 y la edad más frecuente de inicio eran los 19,71 años (AU)