Case of Renal Aspergillosis after Heart Transplant: Diagnosis and Treatment.

Invasive aspergillosis (IA) is a severe complication after heart transplantation (HTx), with a high mortality rate. Invasive pulmonary aspergillosis (IPA) is the most common presentation. We herein describe a unique case of Renal aspergillosis (RAsp) diagnosed on month 10 post-HTx with no known risk factors for IPA in cardiac transplant recipients. The diagnosis of RAsp was made based on radiographic findings, renal biopsy, and tissue cultures. The patient initially received combined antifungal therapy (caspofungin and voriconazole) without radical or partial nephrectomy, followed by voriconazole maintenance monotherapy with favorable clinical outcomes.

Calprotectin (S100A8/S100A9)-induced cytotoxicity and apoptosis in human gastric cancer AGS cells: Alteration in expression levels of Bax, Bcl-2, and ERK2.

Calprotectin is a heterodimeric EF-hand Ca2+ binding protein that is typically released by infiltrating polymorphonuclear leukocytes and macrophages. This protein is a key player linking inflammation and cancer. Due to the increased levels of calprotectin in different inflammatory diseases and cancer, it is considered as a marker for diagnostic purposes. In this study, we evaluated the mechanism of cell viability and apoptotic-inducing effects of recombinant human calprotectin (rhS100A8/S100A9) on the gastric adenocarcinoma (AGS), the most common type of gastric cancer cell line. AGS cells were exposed to the different concentrations (5-100 µg/ml) of calprotectin for 24, 48, and 72 h, and cell viability was assessed through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Apoptotic-inducing effects of calprotectin were evaluated by sub-G1 cell cycle assay and Annexin V/propidium iodide double staining. Furthermore, real-time polymerase chain reaction and Western blot analysis were performed to evaluate the mechanism of action of calprotectin. Our findings indicated that calprotectin inhibits growth and viability of AGS cells in a time- and dose-dependent manner. The half-maximal inhibitory concentration values were measured as 85.77, 79.14, and 65.39 µg/ml for 24, 48, and 72 h, respectively. Additionally, we found that calprotectin downregulated the expression of antiapoptotic protein Bcl-2 and upregulated proapoptotic protein Bax in a time- and concentration-dependent fashion. Calprotectin also slightly upregulated the expression of extracellular signal-regulated protein kinase 2 (ERK2), while it significantly decreased the levels of phospho-ERK in a time-dependent manner. Overall, these findings indicated that calprotectin has cytotoxicity and apoptosis-inducing effects on AGS cell lines in high concentration by modulating Bax/Bcl-2 expression ratio accompanied by inhibition of ERK activation.

2-NDC from dithiocarbamates improves ATRA efficiency and ROS-induced apoptosis via downregulation of Bcl2 and Survivin in human acute promyelocytic NB4 cells.

Although it has been widely considered that all-trans retinoic acid (ATRA) is an efficient therapeutic agent for acute promyelocytic leukemia (APL), there is an urgent need for extending and examining new therapeutics in medicine. Dithiocarbamates (DTCs) are one of the recent important chemical synthetic compounds used in cancer therapy. The aim of this study was to evaluate the apoptosis-inducing effect of 2-nitro-1-phenylethylpiperidine-1-carbodithioate (2-NDC) as an active derivative from DTCs, in combination with ATRA on human APL NB4 cells. The viability of treated NB4 cells was measured by 3-(4,5-dimethyltiazol-2-yl)-2,5-diphenyltetrazolium bromide assay in various concentrations (10-120 µM). The proapoptotic effects of 2-NDC were investigated by acridine orange/ethidium bromide staining, DNA ladder formation, and flow cytometry. We also assessed the oxidative stress-inducing effect of 2-NDC and in combination with ATRA on the NB4 cells. The alteration in gene expression levels of Bax, Bcl2, and Survivin was measured through a real-time polymerase chain reaction. Furthermore, we redetected the interaction between 2-NDC and antiapoptotic proteins Bcl2 and Survivin via molecular docking. We found that 2-NDC induced apoptosis in NB4 cells in a time-dosage-dependent manner. Also, 2-NDC triggered apoptosis by expanding intracellular reactive oxygen species, combined with ATRA. Bax/Bcl2 ratio was modulated and Survivin was downregulated in NB4 cells upon 2-NDC treatment. Molecular docking studies indicated that 2-NDC binds to the baculovirus inhibitor of apoptosis protein repeat domain of Survivin and Bcl homology 3 domain of Bcl2 with various affinities. Based on the present observations, it seems that this derivative can be estimated as an appropriate candidate for future pharmaceutical evaluations.

Apoptosis-inducing effect of spiroaminopyrimidine analogue in NB4 leukemia cells via down-regulation of BIRC5 expression.

AIM: It has been reported that spiroaminopyrimidine derivatives inhibited the growth and proliferation of various cancer cell lines. In the present study, we evaluated cytotoxic and apoptosis-inducing effects of 2,4-diamino-1,3-diazaspiro[5.5]-9-tert-butyl-2, 4-diene-5-carbonitril (9-tBAP) on NB4 acute promyelocytic leukemia (APL) cells. MATERIALS AND METHODS: The cells were treated with 10-100 µM of 9-tBAP. Cytotoxic activity of the compound was measured using the MTT assay. Apoptosis was investigated by Hoechst 33258 staining as well as by Annexin V/PI double staining. RESULTS: The compound under study was found to be highly active cell growth inhibitor with IC50 of 30.0 ± 3.5 µM inducing apoptosis in NB4 cells. Cell cycle analysis by flow cytometry showed a time-dependent increase in sub-G1 cell population. Real-time polymerase chain reaction analysis revealed that the treatment with the compound down-regulated the BIRC5 expression in a time-dependent manner. CONCLUSION: 9-tBAP displayed potent anti-leukemic activity in vitro thus warranting further investigation.

Systemic infection with Candida albicans in breast tumor bearing mice: Cytokines dysregulation and induction of regulatory T cells.

OBJECTIVE: The effect of candidemia on immunologic parameters in breast tumor bearing patients is not well studied. Here, we hypothesised that candidemia in the tumor background may change the outcome of immunologic parameters and tumor condition. METHOD: Mice were divided into four groups, including normal, tumor, Candida infected (only Candidiasis) and tumor/Candidiasis groups. Tumor changes were recorded daily after tumor transplantation and induction of candidemia. Splenocytes of mice were harvested, cultured, and stimulated with PHA; afterwards, IL-4, IL-10, IFN-γ, TNF-α and TGF-ß cytokines were assessed using ELISA kits. We also evaluated the population of CD4+CD25+Foxp3+ regulatory T cells in the tumor infiltrated and splenocytes. RESULTS: The results showed that infection with C. albicans decreased the IFN-γ/IL-4 ratio in tumor/candidiasis and candidiasis groups versus their non-infected controls. IL-10, TGF-ß and TNF-α levels increased in the candidiasis group. In addition, Candidemia led to an increase in the Treg population in tumor microenvironment and splenocytes of experimental groups compared with non-infected controls. Finally, candidemia increased tumor growth of tumor/Candidiasis group compared with the tumor group. CONCLUSION: It seems that systemic infection with C. albicans could not only induce regulatory T cells but also result in dysregulation of cytokine network and thereby facilitate tumor growth.

Socio-Economic Status and Prevalence of Self-Reported Osteoporosis in Tehran: Results from a Large Population-Based Cross-Sectional Study (Urban HEART-2).

Osteoporosis is a widespread disease among older peoples. The aim of this study is to estimate the prevalence of self-reported osteoporosis and assessing its association with socio-economic status. A population-based cross-sectional study was conducted in Tehran, Iran in 2011. Participants were 45,990 individuals aged above 20 years from 22 urban districts. Osteoporosis was measured by self-administrative questionnaire. Wealth index was constructed using principal component analysis based on household assets. Chi-square test, chi square test for trend, and crude odds ratio were used to assess associations in univariate analysis. Multiple logistic regression utilized to estimate adjusted associations between self-reported osteoporosis and socio-economic status.The overall estimated prevalence of self-reported osteoporosis was 4% (95% CI 3.88-4.13), 1.19% in men, and 6.84% in women (P < 0.001). The prevalence increased considerably as age increased (P for trend < 0.001). In multivariable analysis, education and wealth status were negative, and smoking was positively associated with the prevalence of self-reported osteoporosis. No association was found between participants' skill levels and Townsend deprivation index with the prevalence of self-reported osteoporosis.The findings of the present study have improved understanding of the association between socioeconomic status and osteoporosis in the Iranian population. It is important to consider socioeconomic status in screening and prevention programs.

Cytotoxicity, oxidative stress, and apoptosis in K562 leukemia cells induced by an active compound from pyrano-pyridine derivatives.

Recent studies have reported the potential of pyrano-pyridine compounds in inhibiting cell growth and apoptosis induction in cancer cells. Here, we investigated the effect of new pyrano-pyridine derivatives on proliferation, oxidative damages, and apoptosis in K562 leukemia cells. Among different tested compounds, we found 8-(4-chlorobenzylidene)-2-amino-4-(4-chlorophenyl)-5, 6, 7, 8-tetrahydro-6-phenethyl-4H-pyrano-[3,2-c]pyridine-3-carbonitrile (4-CP.P) as the most effective compound with IC50 value of 20 µM. Gel electrophoresis, fluorescence microscopy, and flow cytometry analyses indicated the apoptosis induction ability of 4-CP.P in K562 cells. Further analyses revealed that 4-CP.P induces significant increase in cellular reactive oxygen species production, lipid peroxidation, protein oxidation, and total thiol depletion. Interestingly, while 4-CP.P significantly increased the activity of superoxide dismutase, it reduced the catalase activity in a time-dependent manner. These data propose that 4-CP.P treatment causes free radicals accumulation that ultimately leads to oxidative stress condition and apoptosis induction. Therefore, we report the 4-CP.P as a novel, potent compound as a chemotherapeutic agent in cancer treatment.

Analysis of the cox1 gene in Echinococcus granulosus from sheep in northeast Iran using PCR high-resolution melting (qPCR-HRM) curve analysis.

Echinococcus granulosus, the etiologic agent of echinococcosis, is one of the most important zoonotic helminthes worldwide. Knowledge of E. granulosus species and genotypes has important implications for epidemiology, control, and prevention of diseases as well as future vaccine and drug designs. There are many molecular methods developed to define genotypes of E. granulosus, among them high resolution melting (HRM) analysis, as a new approach, is a single step and closed tube method. It is appropriate for fast screening of large number of isolates. This technique is an accurate, user friendly, cost-effective, fast and simple method, which does not need post-PCR processes. Between March and lst august 2016, of 726 sheep examined in abattoirs in Razavi Khorasan province, Northeast Iran, 109 harboured cystic echincoccosis lesions (liver samples= 65 and lung samples= 44) which were collected for analysis. Total genomic DNA was extracted from each sample and amplified for the presence of polymorphism in the mitochondrial cox1 gene of Echinococcus granulosus using a high resolution melting curve (HRM) method. A total of 109 hydatid cyst samples analyzed by PCR high-resolution melting (qPCR-HRM) curve of the cox1 gene, all isolates were identified as G1 genotype (sheep strain). G1 is the predominant genotype in sheep in northeast of Iran. The high incidence of the G1 genotype (known to be the predominant E. granulosus genotype infecting humans globally) in sheep has considerable implications for hydatid disease control programs in this area.

Combined effect of aerobic interval training and selenium nanoparticles on expression of IL-15 and IL-10/TNF-α ratio in skeletal muscle of 4T1 breast cancer mice with cachexia.

Cancer cachexia is characterized by inflammation, loss of skeletal muscle and adipose tissue mass, and functional impairment. Oxidative stress and inflammation are believed to regulate pathways controlling skeletal muscle wasting. The aim of this study was to determine the effects of aerobic interval training and the purported antioxidant treatment, selenium nanoparticle supplementation, on expression of IL-15 and inflammatory cytokines in 4T1 breast cancer-bearing mice with cachexia. Selenium nanoparticle supplementation accelerated cachexia symptoms in tumor-bearing mice, while exercise training prevented muscle wasting in tumor-bearing mice. Also, aerobic interval training enhanced the anti-inflammatory indices IL-10/TNF-α ratio and IL-15 expression in skeletal muscle in tumor-bearing mice. However, combining exercise training and antioxidant supplementation prevented cachexia and muscle wasting and additionally decreased tumor volume in 4T1 breast cancer mice. These finding suggested that combining exercise training and antioxidant supplementation could be a strategy for managing tumor volume and preventing cachexia in breast cancer.

Synthesis and characterization of Fe3O4-PEG-LAC-chitosan-PEI nanoparticle as a survivin siRNA delivery system.

The limited effectiveness of the conventional methods for cancer treatment makes the researchers to find novel safe and effective therapeutic strategies. One of these strategies is to use small interfering RNAs (siRNAs). A major challenge here is the siRNA delivery into the cells. The purpose of this study was to design and prepare a biocompatible, biodegradable, and safe nanosized particle for siRNA delivery into human breast cancer MCF-7 and leukemia K562 cells. Chemically synthesized magnetic nanoparticles containing polyethyleneglycol-lactate polymer (PEG-LAC), chitosan, and polyethyleneimine (PEI) were successfully prepared and used as a gene delivery vehicle. The nanoparticles were characterized by Fourier transform infrared spectroscopy and zeta potential. The Fe3O4-PEG-LAC-chitosan-PEI nanoparticle showed efficient and stable survivin siRNA loading in gel retardation assay. The cytotoxicity of the prepared nanoparticle was studied using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay and was compared with that of mitoxantrone (MTX) in combination with the prepared siRNA delivery system to evaluate the possible synergic effect of MTX and survivin siRNA. The nanoparticles with and without noncomplementary siRNA showed low toxicity against both cell lines; however, a twofold decrease was observed in cell survival percent after MTX addition to MCF-7 cells treated with either nanoparticle itself or complexed with noncomplementary siRNA. While survivin siRNA nanoplex caused threefold decrease in the cell survival percent, its combination with MTX did not result in a significant increase in the cytotoxic effect. Therefore, Fe3O4-PEG-LAC-chitosan-PEI nanoparticle should be considered as a potential carrier for enhanced survivin siRNA delivery into MCF-7 and K562 cells.

Expression of interleukin-15 and inflammatory cytokines in skeletal muscles of STZ-induced diabetic rats: effect of resistance exercise training.

Skeletal muscle atrophy is associated with type-1 diabetes. Skeletal muscle is the source of pro- and anti-inflammatory cytokines that can mediate muscle hypertrophy and atrophy, while resistance exercise can modulate both muscle mass and muscle cytokine expression. This study determined the effects of a 5-week resistance exercise training regimen on the expression of muscle cytokines in healthy and streptozotocin-induced diabetic rats, with special emphasis on interleukin-15 (IL-15), a muscle-derived cytokine proposed to be involved in muscle hypertrophy or responses to stress. Induction of diabetes reduced muscle weight in both the fast flexor hallucis longus (FHL) and slow soleus muscles, while resistance training preserved FHL muscle weight in diabetic rats. IL-15 protein content was increased by training in both FHL and soleus muscles, as well as serum, in normal and diabetic rats. With regard to proinflammatory cytokines, muscle IL-6 levels were increased in diabetic rats, while training decreased muscle IL-6 levels in diabetic rats; training had no effect on FHL muscle IL-6 levels in healthy rats. Also, tumor necrosis factor-alpha (TNF-α) and IL-1ß levels were increased by diabetes, but not changed by training. In conclusion, we found that in diabetic rats, resistance training increased muscle and serum IL-15 levels, decreased muscle IL-6 levels, and preserved FHL muscle mass.

Investigating the altitude effect on the quantity and quality of the essential oil in Tanacetum polycephalum Sch.-Bip. polycephalum in the Baladeh region of Nour, Iran.

Medicinal plant are grown and produced in different ecosystems and sites under the influence of different potential factors, including the altitude as one of the vital determinants in the quantity and quality of the plants. One of the species that grows in the highlands is Tanacetum polycephalum, an aromatic perennial of the Asteraceae. This species is characterized to be antiseptic, analgesic, anesthetic, disinfective, expectorant, anti-cancer, anti-allergic, and conducive to low blood pressure. The purpose of this study is to investigate the essential compositions in the aerial parts of T. polycephalum at the time of flowering, and in three different altitudes of the Baladeh region of Nour. Thus, the essential oil was extracted from the aerial parts in the flowering stage of the plant at three altitudes of 1 600, 2 400 and 3 200 m using a water distillation method, and the essential oil compositions were identified using GC and GC/MS instruments. One-way ANOVA method was conducted to analyze the obtained data using SPSS, and a Duncan test was administered to compare the means. The results indicated that the essential output obtained from the altitudes of 1 600, 2 400 and 3 200 m was (0.74 ± 0.01)%, (1.09 ± 0.02)%, and (1.32 ± 1.2)%, respectively, so that the altitude of 3 200 m revealed the greatest quantity, and the altitude of 1 600 m represented the smallest quantity. Moreover, the essential oil compositions showed the highest percentage in the altitude of 3 200 m and the lowest percentage at the altitude of 1 600 m. The results showed that as the altitude increases, the essential oil compositions revealed the greater quantity and percentage in the aerial parts of T. polycephalum.

The preventive oral supplementation of a selenium nanoparticle-enriched probiotic increases the immune response and lifespan of 4T1 breast cancer bearing mice.

The immunomodulatory effects of lactic acid bacteria have been demonstrated previously. In this study, a Lactobacillus plantarum strain was selected and enriched with selenium nanoparticles for use as a new immunomodulating agent in a breast cancer murine model. 30 female inbred BALB/c mice were equally divided into a test group and a control group. For 2 weeks prior to tumor induction, each mouse received a daily oral administration of 2.5×108 CFU/ml of L. plantarum enriched with selenium nanoparticles (SeNPs), and then 1×106 4T1 cells were injected subcutaneously. After tumor induction, daily SeNP administration was repeated for 3 cycles of 7 days on/3 days off. Immunological parameters such as levels of cytokines, NK cell activity, tumor growth, and mouse survival were evaluated. The production of the pro-inflammatory cytokines IFN-γ, TNF-α, and IL-2 in spleen cell cultures was increased in test mice administered SeNP-enriched L. plantarum. The test mice also showed significant increases in NK cell activity. The tumor volumes of treated mice were decreased and their survival rate notably increased when compared to mice that received L. plantarum alone or control mice. Administration of SeNP-enriched L. plantarum can induce an efficient immune response through the elevation of the pro-inflammatory cytokines IFN-γ, TNF-α and IL-2 levels and increased NK cell activity. Therefore, this treatment may result in better cancer prognosis.

Vertical strabismus cases and their surgical outcome in Yazd, Islamic Republic of Iran.

Early diagnosis and management of strabismus is needed to avoid complications such as amblyopia. In this retrospective cohort study we reviewed the profile and surgical outcome of vertical strabismus patients attending a clinic in Yazd city in the Islamic Republic of Iran. From the medical files of 265 patients, 19.2% were found to have pure vertical deviation and 80.8% had combined horizontal and vertical strabismus. Congenital causes were recorded for 83.0% of cases and 24.9% had a family history of strabismus. Hypertropia (91.7%) was more common than hypotropia (8.3%). There was a statistically significant relationship between age at onset and vertical strabismus. Female patients had a higher rate of congenital vertical strabismus than did male patients. There was no significant association between surgery outcomes and sex or age of onset of strabismus. Early diagnosis and management in affected families and screening ofpatientswith strabismus family history is needed.

Credibility of the measurement of serum ferritin and transferrin receptor as indicators of iron deficiency anemia in hemodialysis patients.

BACKGROUND AND OBJECTIVES: Anemia is a common complication in uremic patients. Erythropoietin therapy is prescribed in these cases; however, this treatment is not successful in iron deficient patients. Ferritin-based diagnosis of iron deficiency in these patients is a challenging task, as serum ferritin level may be high due to chronic inflammation and mask iron deficiency. In the current study we evaluated the credibility of another indicator of body iron supply, serum transferrin receptor, in hemodialysis patients in two University-based Hospitals in North of Iran. MATERIALS AND METHODS: In a cross-sectional study, 53 hemodialysis patients with a mean age of 56 +/- 18.7 years and 30 persons with iron deficiency and normal renal function with a mean age of 20.1 +/- 14.4 years were examined. All hemodialysis patients were on hemodialysis 2-3 times per week for 3-4 hours. All cases were examined for blood hemoglobin content, serum iron, CRP, serum ferritin and serum transferrin receptor levels. The reference ranges introduced by manufacturers were considered as standard ranges for analysis of the results. Using one sample T-test and Fisher's exact test, data were analyzed. p<0.05 was considered as significant. RESULTS: Hemodialysis patients had blood hemoglobin content below normal range (p<0.05 for men, p<0.001 for women) and CRP levels above normal range (p<0.001). In hemodialysis patients, serum ferritin level was significantly higher than control group (p<0.001), whilst serum transferrin receptor levels in the two groups were not significantly different (p=0.69), and both were above defined normal upper limit (p<0.001 for iron deficient patients; p<0.05 for hemodialysis patients). DISCUSSION: This study showed measurement of serum ferritin in the presence of chronic inflammation induced by renal failure cannot be a credible indicator of body iron supply, while under this certain condition serum transferrin receptor can more appropriately reflect the amount of body iron supply.

Synthesis and cytotoxic activity of some 2-amino-4-aryl-3-cyano-7-(dimethylamino)-4H-chromenes.

A series of 2-amino-4-aryl-3-cyano-7- (dimethylamino)-4H-chromenes was synthesized by condensation of 3- (dimethylamino) phenol, an aromatic aldehyde and malonitrile in ethanol containing piperidine. The assignments of the structure of all synthesized compounds were based on spectral data (IR, Mass and(1)H NMR). The cytotoxic activities of the synthesized compounds against six human tumor cell lines were determined by MTT assay. Several compounds showed significant cytotoxic activity.

Preparation of antibody against horseradish peroxidase using hybridoma technology.

BACKGROUND: Monoclonal antibody against horseradish peroxidase (HRP) has many applications which peroxidase anti-peroxidase. Peroxidase-antiperoxidase (PAP) complex formation is its most known and important usage. This complex is used in many immunohistochemical and immunocytochemical staining techniques. OBJECTIVE: The aim of this study was the preparation of anti-HRP monoclonal antibody through hybridoma technology. METHODS: The BALB/c mice were immunized by repeated injections of HRP. After the confirmation of their immunization by ELISA test, the spleen lymphocytes and SP2/0 myeloma cells were hybridized using PEG as fusing agent. The hybridoma cells were then selected by culturing in HAT medium. Identification and selection of anti-HRP producing clones were done by ELISA test on culture supernatants of the obtained clones. To acquire the monoclones, limiting dilution was performed twice and the effect of finally obtained antibodies on enzyme activity was investigated by a specific ELISA test. In vivo tumor induction method was used for production of concentrated antibody. At last class and subclass of the obtained antibodies were determined by Isostrip Kit. RESULTS: After seven rounds of cell fusions, 224 clones were obtained, from which, six ones were anti-HRP producers. Two clones (P1F11 and P2F6) with higher antibody secretion were selected and subcloned. Both derived hybridoma monoclones (P1F11D2 and P2F6F3) were producing antibodies from IgG1 subclass with kappa (Kappa) light chains which didn't affect the enzyme activity. The electrophoresis of ascetic fluid of tumor induced mice showed an obvious band in gamma (gamma) position. CONCLUSION: The obtained monoclonal antibodies are from IgG class and don't affect the enzyme activity, therefore it seems that they are suitable for PAP complex production.

Unwanted pregnancy among kidney transplant recipients in Iran.

To investigate the incidence of unwanted pregnancy among kidney transplant recipients, we studied 86 pregnancies in 64 women with a transplanted kidney. Twenty-five pregnancies were unwanted (29.1%). Pregnancy was terminated by induced abortion in seven patients, and four pregnancies were lost due to spontaneous abortion with one intrauterine fetal death. Only 13 (52%) pregnancies resulted in a live birth. Most of the unwanted pregnancies occurred in women using coitus interruptus (92%) as the only method of contraception. It is concluded that because fertility greatly improves after kidney transplantation, it is necessary to have a family planning counseling session before surgery. If a patient is not interested in future pregnancy, an effective method of contraception should be offered. A woman who has decided against childbearing in the future may decide to have a tubal ligation at the time of transplantation surgery.

High incidence of autosomal recessive nonsyndromal congenital retinal nonattachment (NCRNA) in an Iranian founding population.

In an isolated and founding Iranian population the prevalence of congenital total blindness is 1.1%. Clinical findings such as lack of perception of light, massive retrolental mass, shallow anterior chamber and nystagmus, in otherwise normal individuals, correspond to nonsyndromal congenital retinal nonattachment. To determine the inheritance of this disease we constructed an extensive nine-generation pedigree of the affected kindred living in the Iranian founding population. The pedigree, which includes 42 patients from 25 sibships, clearly suggests autosomal recessive inheritance. To verify the inheritance, we compared the average coefficient of inbreeding (F) of the affected sibships with that of the control sibships, calculated the patients' sex ratio, and also compared the observed relative frequency of the disease with its expected relative frequencies for different modes of inheritance. The patients' average F value is significantly greater than that of the controls (P < 0.001). The sex ratio of the patients is close to unity and the observed relative frequency of the disease is close to that of an autosomal recessive trait. All these findings strongly support autosomal recessive transmission of this disease.