An intrafamilial phenotypic variability in Ellis-Van Creveld syndrome due to a novel 27 bps deletion mutation.

Ellis-van Creveld (EvC) syndrome is an autosomal recessive disease, characterized by ectodermal, skeletal, and cardiac anomalies. We report intrafamilial phenotypic variability in three new EvC syndrome cases. Affected males in this study showed only ectodermal abnormalities, whereas an affected female showed the classical presentation of EvC Syndrome, including bilateral postaxial polydactyly of hands and feet, and congenital heart defects. Whole exome sequencing was performed to identify the causative variant, followed by validation and segregation analysis using Sanger sequencing. A homozygous deletion variant (c.731_757del) was identified in exon 6 of the EVC gene (NM_153717.2). The identified variant is considered to be the most likely candidate variant for the EvC syndrome in the family based on previous reports validating the role of EVC variants in the EvC syndrome. The disease correctly segregated in the family members, as all affected members were homozygous, and obligate carriers were heterozygous. Our family is remarkable in highlighting the variable expressivity of the EvC phenotype within the same family, due to a homozygous deletion mutation in the EVC gene. The variable expressivity might be due to the hypomorphic nature of mutation, or the presence of additional variants in modifier genes or in the regulatory regions of the EVC/EVC2 genes.

Expression and in vitro anticancer activity of Lp16-PSP, a member of the YjgF/YER057c/UK114 protein family from the mushroom Lentinula edodes C91-3.

Latcripin-16 (Lp16-PSP) is a gene that was extracted as a result of de novo characterization of the Lentinula edodes strain C91-3 transcriptome. The aim of the present study was to clone, express, and investigate the selective in vitro anticancer potential of Lp16-PSP in human cell lines. Lp16-PSP was analyzed using bioinformatics tools, cloned in a prokaryotic expression vector pET32a (+) and transformed into E. coli Rosetta gami. It was expressed and solubilized under optimized conditions. The differential scanning fluorometry (DSF)-guided refolding method was used with modifications to identify the proper refolding conditions for the Lp16-PSP protein. To determine the selective anticancer potential of Lp16-PSP, a panel of human cancerous and non-cancerous cell lines was used. Lp16-PSP protein was identified as endoribonuclease L-PSP protein and a member of the highly conserved YjgF/YER057c/UK114 protein superfamily. Lp16-PSP was expressed under optimized conditions (37 °C for 4 h following induction with 0.5 mM isopropyl ß-D-1-thiogalactopyranoside). Solubilization was achieved with mild solubilization buffer containing 2 M urea using the freeze-thaw method. The DSF guided refolding method identified the proper refolding conditions (50 mM Tris-HCl, 100 mM NaCl, 1 mM EDTA, 400 mM Arginine, 0.2 mM GSH and 2 mM GSSG; pH 8.0) for Lp16-PSP, with a melting transition of ~ 58 °C. A final yield of ~ 16 mg of purified Lp16-PSP from 1 L of culture was obtained following dialysis and concentration by PEG 20,000. A Cell Counting Kit-8 assay revealed the selective cytotoxic effect of Lp16-PSP. The HL-60 cell line was demonstrated to be most sensitive to Lp16-PSP, with an IC50 value of 74.4 ± 1.07 µg/ml. The results of the present study suggest that Lp16-PSP may serve as a potential anticancer agent; however, further investigation is required to characterize this anticancer effect and to elucidate the molecular mechanism underlying the action of Lp16-PSP.

Hg isotopic composition and total Hg mass fraction in NIES Certified Reference Material No. 28 Urban Aerosols.

An interlaboratory study on the National Institute for Environmental Studies (NIES) certified reference material (CRM) No. 28 Urban Aerosols, collected on the filters of a central ventilating system in a building in Beijing city center, was performed to obtain informative values of Hg isotopic composition and total Hg (THg) mass fraction. The THg mass fraction was determined by four organizations using atomic absorption spectrometry; it resulted in the mean value of 1.19 ± 0.12 mg/kg (2SD, n = 24). The Hg isotopic composition of the CRM was measured and intercompared at two different institutions by cold vapor generation system coupled to multicollector inductively coupled plasma mass spectrometry. Subsequently, a conventional dissolution method that uses a mixture of HNO3/HCl/H2O2 in Hotblock® and two different dissolution methods that use a mixture of HNO3/HCl with a microwave and a digestion bomb were applied. The Hg isotopic compositions were δ202Hg = - 1.26 ± 0.17‰, Δ199Hg = - 0.23 ± 0.06‰, Δ200Hg = 0.01 ± 0.07‰, and Δ201Hg = - 0.22 ± 0.09‰ (2SD, n = 18) for the conventional method, which agree well with those obtained using microwave and bomb digestion. Our results indicate that, for the quality control of particulate matter analyses, this CRM is appropriate for use in environmental and geochemical studies. Graphical abstract.

Latcripin-7A, derivative of Lentinula edodes C91-3, reduces migration and induces apoptosis, autophagy, and cell cycle arrest at G1 phase in breast cancer cells.

Due to the high mortality rate and an increase in breast cancer incidence, it has been challenging for researchers to come across an effective chemotherapeutic strategy with minimum side effects. Therefore, the need for the development of effective chemotherapeutic drugs is still on the verge. Consequently, we approached a new mechanism to address this issue. The naturally available peptide named latcripin-7A (LP-7A), extracted from a mushroom called Lentinula edodes, provided us promising results in terms of growth arrest, apoptosis, and autophagy in breast cancer cells (MCF-7 and MDA-MB-231). Expressions of protein markers for apoptosis, autophagy, and cell cycle were confirmed via Western blot analysis. Migration and invasion assays were performed to analyze the anti-migratory and anti-invasive properties of LP-7A, while cell cycle analysis was performed via flow cytometry to evaluate its affect over cell growth. Supportive assays were performed like acridine orange, Hoechst 33258 stain, DNA fragmentation, and mitochondrial membrane potential (MMP) to further confirm the anticancer effect of LP-7A on breast cancer cell lines. It is concluded that LP-7A effectively reduces migration and promotes apoptosis as well as autophagy in MCF-7 and MDA-MB-231 breast cancer cell lines by inducing cell growth arrest at G0/G1 phase and decreasing mitochondrial membrane potential without adverse effects on MCF-10A normal breast cells. KEY POINTS: • In this study, we have investigated the anti-cancer activity of novel latcripin-7A (LP-7A), a protein extracted as a result of de novo characterization of Lentinula edodes C91-3. • We conclude in our research work that LP-7A can initiate diverse cell death-related events, i.e., apoptosis and autophagy in both triple-positive and triple-negative breast cancer cell lines by interacting with different nodes of cellular signaling that can further be investigated in vivo to gain a better understanding.

Enhanced antimicrobial, antioxidant, in vivo antitumor and in vitro anticancer effects against breast cancer cell line by green synthesized un-doped SnO2 and Co-doped SnO2 nanoparticles from Clerodendrum inerme.

A novel approach was employed for the synthesis of un-doped tinoxide and Cobalt-doped tinoxide (Co-doped SnO2) nanoparticles (NAPs) by using aqueous extract of Clerodendrum inerme with the help of eco-friendly superficial solution combustion method. Synthesized NAPs were characterized by different spectroscopic techniques and results from XRD, TEM, SEM, EDX and UV-Vis examines confirmed the successful synthesis, crystalline nature and spherical structure of un-doped SnO2 and Co-doped SnO2 NAPs with the average grain size of 30 and 40 nm; and band gap energy of 3.68 and 2.76 eV respectively. Antimicrobial propensity of the synthesized NAPs was determined by agar well assay, SEM, TEM and confocal laser scanning microscopic analysis against various bacterial and fungal strains. Synthesized Co-doped SnO2 NAPs were unveiled the extraordinary antibacterial and antifungal activities against E. coli, B. subtilis, A. niger, A. flavus, and C. albicans with the zone of inhibitions of 30 ±â€¯0.08 mm and 26 ±â€¯0.06 mm, 17 ±â€¯0.04 mm, 23 ±â€¯0.08 mm and 26 ±â€¯0.06 respectively which were also evidenced from SEM, TEM and confocal laser scanning microscopy. In addition, green synthesized Co-doped SnO2 NAPs were demonstrated the substantial antioxidant activity by scavenging DPPH, significant in vitro anticancer and in vivo antitumor activity on breast carcinoma cells (MCF-7) and Ehrlich ascites tumor cell lines respectively than standard. The hemolytic activity disclosed low cytotoxicity of fabricated NAPs (0.89 ±â€¯0.05%) at 5 mg/mL, which was indicated their biocompatibility potential. Hence, the multi-purpose properties of synthesized NAPs presented in the current study can be further deliberated for pharmaceutical and nanomedicine applications.

Assessment of physicochemical parameters of wastewater samples.

Water samples from selected locations of Nullah Lai and Koh-e-Noor textile mill in the metropolitan city of Rawalpindi and Islamabad, Pakistan, were collected. Physicochemical parameters and heavy metals were determined using standard analytical procedures in comparison with sites, locations and subsequent interval of 3 months. The results of the physicochemical analysis at different locations of Nullah Lai and Koh-e-Noor textile mill with an interval of 3 months were obtained in the following range: pH (7.16-8.29), temperature (17.8-28.8 °C), conductivity (1,005-3,347 µS/m), TDS (754.3-2,519.5 mg/L), turbidity (272.8-487.05 NTU), total hardness (300-452 mg/L), nitrates (10.11-22.95 ppm), calcium (74.31-139.2 ppm), chloride (127.72-396.16 ppm), sulphate (15.97-87.38 ppm), NaCl (210.5-631.1 ppm), Ni (0.30-0.72 ppm), Cd (0.005-0.03 ppm), Cr (0.2-7.4 ppm), Pb (0.12-0.73 ppm), Zn (0.03-0.08 ppm) and Cu (0.01-0.06 ppm). The highest value of physicochemical parameters (compared with Nullah Lai) was obtained in locations of Koh-e-Noor textile mill. The results obtained exceeded the maximum allowable limit set by the World Health Organization for drinking purpose but can be used for irrigation purposes after suitable treatment and purification.

Characterization of Green and Yellow Papaya (Carica papaya) for Anti-Diabetic Activity in Liver and Myoblast Cells and Wound-Healing Activity in Fibroblast Cells.

Obesity and diabetes, often characterized as "metabolic syndrome", have been recognized as two of the most important public health issues worldwide. The objective of the present research was to evaluate green and yellow papaya for anti-oxidation and anti-diabetic properties. Leaves, skin, pulp, and seed samples from papayas were freeze-dried and then extracted in water or 80% methanol. The extracts were used to determine total polyphenolic content and anti-oxidation activities, and to determine biological activities, including glucose uptake, Glut-2 expression, triglyceride reduction, and wound-healing activity. Our data demonstrated that methanol and water extracts of green and yellow papaya have similar concentrations of polyphenols in skin (10-20 mg/g dry powder), leaf (25-30 mg/g dry powder), and pulp (1-3 mg/g dry powder) fractions. However, both methanol and water extracts of seeds from yellow papaya have substantially higher concentrations of polyphenols compared to green papaya. Both water and methanol extracts of yellow papaya exhibited higher anti-oxidation activity compared to green papaya in skin (50-60%), pulp (200-300%), and seeds (10-800%). Old leaves also showed greater anti-oxidation activity (30-40%) compared to new leaves. Pulp extracts from both yellow and green papaya stimulated greater glucose uptake, but only pulp from green papaya stimulated glucose uptake in muscle cells. Similarly, pulp extract stimulated glucose transporter Glut-2 expression in liver cells. The skin, pulp, and seeds of green or yellow papaya showed triglyceride-lowering activity in liver cells by 60-80%, but samples taken from yellow papaya had a more potent effect. Seeds from both green and yellow papaya significantly stimulated the migration of fibroblasts in the wounded area by 2-2.5-fold compared to the untreated control. Consistent with these data, seeds from both green and yellow papaya also significantly stimulated collagen synthesis in fibroblast cells by almost 3-fold. In conclusion, our data indicate that different parts of papaya produce stimulatory effects on glucose uptake, Glut-2 expression, TG reduction, and wound-healing activities. This study concludes that different parts of the papaya can be beneficial for preventing diabetes and diabetes-related wound healing.

Dysregulation of Type I Interferon (IFN-I) Signaling: A Potential Contributor to Racial Disparity in Hepatocellular Carcinoma (HCC).

African-American (AA)/Black hepatocellular carcinoma (HCC) patients have increased incidence and decreased survival rates compared to non-Hispanic (White) patients, the underlying molecular mechanism of which is not clear. Analysis of existing RNA-sequencing (RNA-seq) data in The Cancer Genome Atlas (TCGA) and in-house RNA-sequencing of 14 White and 18 AA/Black HCC patients revealed statistically significant activation of type I interferon (IFN-I) signaling pathway in AA/Black patients. A four-gene signature of IFN-stimulated genes (ISGs) showed increased expression in AA/Black HCC tumors versus White. HCC is a disease of chronic inflammation, and IFN-Is function as pro-inflammatory cytokines. We tested efficacy of ginger extract (GE), a dietary compound known for anti-inflammatory properties, on HCC cell lines derived from White (HepG2), AA/Black (Hep3B and O/20) and Asian (HuH-7) patients. GE exhibited a significantly lower IC50 on Hep3B and O/20 cells than on HepG2 and HuH-7 cells. The GE treatment inhibited the activation of downstream mediators of IFN-I signaling pathways and expression of ISGs in all four HCC cells. Our data suggest that ginger can potentially attenuate IFN-I-mediated signaling pathways in HCC, and cells from AA/Black HCC patients may be more sensitive to ginger. AA/Black HCC patients might benefit from a holistic diet containing ginger.

Effect of sequential or ternary starters-assisted fermentation on the phenolic and glucosinolate profiles of sauerkraut in comparison with spontaneous fermentation.

This study investigated the effectiveness of two novel started-assisted sauerkraut fermentations in comparison with spontaneous fermentation. Three lactic acid bacteria strains were selected as best starters for sauerkraut processing, based on the complementarity of pro-technological (kinetics of growth and acidification) traits, phenotypic fingerprints through OmniLog Phenotype MicroArray, and phenolics metabolism. The selected strains were applied according to two different fermentation methods based on steering sequential and temporally deferred inoculum of three strains, and ternary simultaneous inoculum. Sequential and ternary starters-assisted fermentations lasted 9 and 7 days, respectively, and were compared to conventional spontaneous fermentation lasting 35 days. Sequential and ternary fermentations resulted in a higher and constant number of lactic acid bacteria compared to spontaneous fermentation, which reflected on the acidification and sugar utilization. Ternary fermentation enhanced phenolic compounds conversion (hydrocaffeic acid, hydroferulic acid, 4-ethyl catechol), ensuring at the same time higher level of aliphatic (glucobrassicanapin) and indole glucosinolates (glucobrassicin) and derivatives(ascorbigen). Short fermentation of sequential and ternary starters-assisted processing caused only slight changes in the sensory profile compared to the spontaneous process, preserved the structural integrity of sauerkraut, and did not affect the colour lightness.

Twin birth weight discordance: associated factors and outcome.

OBJECTIVE: To determine the frequency of twin birth weight discordance and compare the maternal characteristics and neonatal outcome in discordant and concordant twin gestations. STUDY DESIGN: Cross-sectional comparative study. PLACE AND DURATION OF STUDY: MCH Centre, Unit-II, Pakistan Institute of Medical Sciences, Islamabad, from January 2005 to December 2007. METHODOLOGY: All twin gestations delivered at > 28 weeks gestation were included. Birth weight discordance was defined as > 20% twin birth weight difference and two groups were developed accordingly. Prenatal complications, gestational age at delivery, mode of delivery and neonatal outcome were compared using chi-square and Fischer exact test with significance at p < 0.05. RESULTS: Two hundred and fifty three pairs of twins were delivered during the three years. The frequency of birth weight discordance was 19% (48 verses 205 concordant twin pairs). Preterm delivery (68% vs. 25%), pre-labour rupture of membranes (33% vs. 9%) and pregnancy induced hypertension (22% vs. 12%) were significantly more frequent in birth weight discordant compared to concordant gestations. The cesarean section rate was 29% and 21% respectively. Twelve (12%) discordant and 29 (7%) concordant infants required NICU admission. There were 10 intrauterine deaths (10%) and one (1%) neonatal death among the discordant twins while 18 (4%) intrauterine deaths and 10 (2.4%) neonatal deaths occurred in concordant twins, resulting in uncorrected PNMR of 114.5 and 43.9 per 1000 births respectively. CONCLUSION: Twin pregnancies with birth weight discordance are at higher risk of prenatal complications and have less favourable perinatal outcome. Their timely antenatal detection is, therefore, important for closer surveillance and timely delivery.

Anti-Obesity Effect of Dictyophora indusiata Mushroom Polysaccharide (DIP) in High Fat Diet-Induced Obesity via Regulating Inflammatory Cascades and Intestinal Microbiome.

Obesity is a multifactorial metabolic disorder characterized by low-grade chronic inflammation, hyper-permeability of the gut epithelium, and perturbation of the intestinal microbiome. Despite the numerous therapeutic efficacies of Dictyophora indusiata mushroom, its biological activity in alleviating obesity through regulation of the gut microbiota and inflammatory cascades remain obscure. Henceforth, we determined the modulatory impact of D. indusiata polysaccharide (DIP) in the high-fat diet (HFD)-induced obesity mice model. The experimental subjects (BALB/C mice) were supplemented with chow diet (Control group), high-fat diet (HFD group), or HFD along with DIP at a low dose [HFD + DIP(L)] and high dose [HFD + DIP(H)]. Obesity-related parameters, including body weight gain, epididymal adipocyte size, fat accumulation, adipogenic markers, lipogenic markers, inflammatory associated markers, intestinal integrity, and intestinal microbiome, were elucidated. Our findings demonstrated that the oral administration of DIP at low dose partially and at high dose significantly reversed HFD-induced obesity parameters. Furthermore, the body weight, fat accumulation, adipocyte size, adipogenic and liver associated markers, glucose levels, inflammatory cytokines, and endotoxin (Lipopolysaccharide, LPS) levels were reduced considerably. Moreover, the study revealed that DIP treatment reversed the dynamic alterations of the gut microbiome community by decreasing the Firmicutes to Bacteroidetes ratio. These findings led us to infer the therapeutic potential of DIP in alleviating HFD-induced obesity via regulating inflammatory cascades, modulating intestinal integrity and intestinal microbiome community.

Speciation and isotopic characterization of mercury detected at high concentration in Pakistani hair samples.

Speciation and isotopic analyses of mercury (Hg) detected at high concentration (11-107 mg total Hg/kg) in the hair samples of Pakistani subjects were carried out to analyze whether the source of Hg was the skin cream or the soap they used. Liquid chromatography-ICP mass spectrometric speciation analysis revealed that the Hg in the hair was primarily inorganic. This result supported our assumption that skin care products were the source of the high concentration of Hg in hair, which was based on the statistical association found in our previous study because it is known that inorganic Hg compounds are added to some skin care products sold in some countries. Stable isotope ratios of the Hg in the hair samples of 6 subjects and 3 skin cream samples, which were reported to be used by the subjects, were measured by multi-collector ICP mass spectrometry after acid digestion. A comparison of the isotopic compositions of Hg in hair and skin cream in two subjects revealed that the composition in both samples matched in one subject, hence confirming that the products were the source of high Hg concentration in hair. However, the isotopic compositions of Hg in hair and skin cream did not match in the other subject demonstrating the presence of other exposure source(s).

A Polysaccharide Isolated from Dictyophora indusiata Promotes Recovery from Antibiotic-Driven Intestinal Dysbiosis and Improves Gut Epithelial Barrier Function in a Mouse Model.

Despite the tremendous biological activity of polysaccharides from the mushroom Dictyophora indusiata, its role in the restoration of gut microbiota has not yet been explored. The present study aimed to investigate whether D. indusiata polysaccharide (DIP) could modulate the recovery of gut microbiota composition and intestinal barrier function after broad-spectrum antibiotic-driven dysbiosis. Alteration and restoration in the microbial communities were elucidated by the Illumina MiSeq platform. Colon histology, expression of tight-junction associated proteins, and serum/tissue endotoxin and cytokine levels were evaluated. Two-week daily oral administration of clindamycin and metronidazole resulted in reduced bacterial diversity and richness, and perturbed the microbial flora at various taxonomic levels (altered Firmicutes/Bacteroidetes ratio and increased relative abundance of harmful flora (Proteobacteria, Enterococcus, and Bacteroides)), whereas DIP administration reversed the dysbiosis and increased beneficial flora, including Lactobacillaceae (lactic acid-producing bacteria), and Ruminococaceae (butyrate-producing bacteria). In addition, it resulted in the reduction of endotoxemia (through lipopolysaccharides (LPSs)) and pro-inflammatory cytokine (tumor necrosis factor alpha (TNF-α), interleukin 6 (IL-6), and interleukin 1ß (IL-1ß)) levels, with the increased expression of tight-junction associated proteins (claudin-1, occludin, and zonula occludens-1). These findings not only suggested a comprehensive understanding of the protective effects of a DIP in the restoration of gut microbiota but also highlighted its role in the enhancement of gut barrier integrity, reduction of inflammation and lowering of endotoxin levels in mice.

Green synthesis of ZnO and Cu-doped ZnO nanoparticles from leaf extracts of Abutilon indicum, Clerodendrum infortunatum, Clerodendrum inerme and investigation of their biological and photocatalytic activities.

In the present study, green synthesis and determination of the antioxidant, antibacterial, antifungal, anticancer and photocatalytic properties of the resultant Cu-doped zinc oxide nanoparticles (NAPs) were carried out. A superficial method (solution combustion method) was employed for the synthesis of un-doped ZnO NAPs from aqueous extract of Abutilon indicum, and synthesis of Cu-doped ZnO NAPs from aqueous extracts of Clerodendrum infortunatum (M1 NAPs) and Clerodendrum inerme (M2 NAPs). The synthesized un-doped ZnO, M1 and M2 NAPs were characterized by different spectroscopic techniques like Ultraviolet-visible (UV-Vis), Fourier Transform-Infrared (FT-IR), X-ray diffraction (XRD), energy-dispersive X-ray spectroscopy (EDX) and scanning electron microscopy (SEM). The antibacterial and antifungal activities of M1 and M2 NAPs were determined by agar diffusion method, while their antioxidant properties were assessed through DPPH radical scavenging and ferric thiocyanate (FTC) and assays. Under sunlight irradiation, photocatalytic disintegration potential of M1 and M2 NAPs were determined by the degradation of Acid Black 234 dye. Results from FT-IR, XRD, EDX and SEM confirmed successful synthesis, crystalline nature, and spheroid-to-rod-like shapes of un-doped ZnO, M1 and M2 NAPs, with grain sizes of 16.72nm, 17.49nm and 20.73nm; and band gap energies of 3.37eV, 3.36eV and 3.31eV, respectively. The NAPs were good catalysts for effective degradation of Acid Black 234. They also exhibited remarkable antioxidant and anticancer activities. Significant antibacterial activity was shown by M2 NAPs against E. coli, S. aureus, Klebsiella and B. subtilis, with zones of inhibition (ZOIs) of 13±0.09, 14±0.01, 18±0.07 and 20±0.10, respectively. Significant antifungal potential was also produced by M2 NAPs at 5mg with ZOIs of 17±0.07 and 24±0.08 against A. niger and T. harzianum, respectively. These results indicate that aqueous extracts of A. indicum, C. infortunatum and C. inerme are effective reducing agents for green synthesis of un-doped ZnO, and Cu-doped ZnO NAPs with significant antimicrobial, antioxidant, and antifungal potential. Thus they are good candidates for future therapeutic applications.

Psychosocial problems of Pakistani parents of Thalassemic children: a cross sectional study done in Bahawalpur, Pakistan.

BACKGROUND: Thalassemia is a blood disorder passed down through families (inherited) in which the body makes an abnormal form of hemoglobin. This disorder results in excessive destruction of red blood cells, and there is no effective treatment. Patients require lifelong blood transfusion, usually started within 6 to 12 months of birth of patient, which on other hand has its own complications. It is a chronic disease that manifests so early in life that it leads to psychological and social problems for parents . We focused on parents to assess the impact of their child's disease. OBJECTIVE: To determine the psychosocial problems of parents of thalassemic children. METHODS: This cross sectional study was conducted among the parents of thalassemic children attending THALASSEMIA CENTRE, BAHAVAL VICTORIA HOSPITAL (BVH), BAHAWALPUR, PAKISTAN during the year 2011. A self designed questionnaire was used that contained questions regarding psychological and social aspects. Patient Health Questionnaire-9 (PHQ-9) was used to assess the depression of parents of thalassemic children. RESULTS: Of the 100 parents interviewed, the majority were mothers (71%) , with a mean age of 32 ± 8.07 years for both sexes. 29 percent of the parents had moderate to severe depression, 16 percent had sleep disturbances. 56 percent were downgraded by relatives. There was a significant relationship between respondent education and depression (p < 0.05). CONCLUSION: A substantial number of parents have psychosocial problems due to the disease of their child. Parent counseling is needed on regular basis.