Novel KIAA0753 mutations extend the phenotype of skeletal ciliopathies.

The skeletal ciliopathies are a heterogeneous group of disorders with a significant clinical and genetic variability and the main clinical features are thoracic hypoplasia and short tubular bones. To date, 25 genes have been identified in association with skeletal ciliopathies. Mutations in the KIAA0753 gene have recently been associated with Joubert syndrome (JBTS) and orofaciodigital (OFD) syndrome. We report biallelic pathogenic variants in KIAA0753 in four patients with short-rib type skeletal dysplasia. The manifestations in our patients are variable and ranging from fetal lethal to viable and moderate skeletal dysplasia with narrow thorax and abnormal metaphyses. We demonstrate that KIAA0753 is expressed in normal fetal human growth plate and show that the affected fetus, with a compound heterozygous frameshift and a nonsense mutation in KIAA0753, has an abnormal proliferative zone and a broad hypertrophic zone. The importance of KIAA0753 for normal skeletal development is further confirmed by our findings that zebrafish embryos homozygous for a nonsense mutation in kiaa0753 display altered cartilage patterning.

The effect of Ramadan fasting on LH, FSH, oestrogen, progesterone and leptin in pregnant women.

Many pregnant Muslim women fast during Ramadan. Leptin has an important role in the reproductive system and hormones. In this study, FSH, LH, oestrogen, progesterone and leptin were measured in the first, second and fourth week of Ramadan and the second week post-Ramadan, in 30 fasting pregnant women. Data were analysed using repeated measures ANOVA by SPSS. The weight and BMI did not change during the study. A significant change in FSH, oestrogen, progesterone and leptin was observed (p < 0.05). The lowest value of FSH was in the second week of Ramadan. Progesterone increased at the end of Ramadan and the second week after. Oestrogen increased significantly during Ramadan and decreased after Ramadan. A decreasing trend was seen in LH during the Ramadan and 2 weeks after (p < 0.1). Leptin decreased significantly 2 weeks after Ramadan. We found poor weight gain and hypoleptinaemia in pregnant fasted women during the study. Food restriction in pregnant fasted women during Ramadan may induce poor weight gain during pregnancy. These data confirm that Ramadan fasting by pregnant women may have potential risks during pregnancy. We recommend further study to evaluate long-term effects of Ramadan fasting during pregnancy in different countries with different food habits and traditions, to obtain reliable and documented data.

First Report of Hyphodermella rosae Causing Dry Fruit Rot Disease on Plum in Iran.

Plum (Prunus domestica) and peach (P. persica) are widely grown, often in alternate rows with citrus, in the Mazandaran Province of Iran. In June 2011, a dry fruit rot of plum was observed in several production regions in Mazandaran Province (35°47'N, 50°34'E). Initial symptoms at pit-hardening stage appeared as dark brown, circular, necrotic spots from 2 to 5 cm in diameter. They later developed into a dry fruit rot. Severe symptoms occurred during June and July when warm weather (temperature around 28°C) and high relative humidity (RH) (>85%) were present. Marketable yield losses reached 50% to almost 100% in many orchards. To isolate the causal organism, symptomatic fruits were surface disinfested for 1 min in 0.5% active chlorine, washed thoroughly with sterile distilled water, and segments were plated on potato dextrose agar (PDA) amended with 50 mg/liter of streptomycin sulfate and incubated at 25°C for 3 days. The fungus Hyphodermella rosae (Bresadola) Nakasone was consistently isolated (37 isolates from 79 samples) and identified on the basis of morphological characteristics on PDA. Basidiomata were effuse, resupinate, 15 × 10 mm, crustaceous, tubercules small with apical bristles, and light orange to grayish orange. Subhymenium was up to 30 µm thick, composed of vertically arranged, short-celled, nonagglutinated hyphae; subhymenial hyphae were 3 to 4 µm in diameter. Basidiospores were ellipsoid, 7.5 to 8.5 × 4.5 to 5.5 µm (100 determination), and their cell walls were thin, hyaline, and smooth (1). Genomic DNA was extracted from mycelium with a DNA extraction kit (Qiagen, Hilden, Germany) according to the manufacturer's directions and grown on potato dextrose broth for 4 days at 28°C. The rDNA region was amplified with the primers ITS4 (5'-TCCTCCGCTTATTGATATGC-3') and ITS5 (5'- GGAAGTAAAAGTCGTAACAA-3') (4) and the PCR product was sequenced. Nucleotide BLAST analysis of the amplified 627-bp fragment confirmed a 99% similarity with the sequence of H. rosae (GenBank Accession No. JN593086). A pathogenicity test was conducted with isolate MA4099 by placing 5-day-old mycelial plugs grown on PDA at the surface of healthy fruit (n = 6) incubated under >85% RH at 25°C for at least 4 days until the appearance of symptoms, which were similar to those displayed under orchard conditions. Control fruits, inoculated with blocks of PDA plugs, remained intact and symptomless. Reisolation from inoculated fruit samples consistently yielded the inoculated fungus, completing Koch's postulates. The genus Hyphodermella has been reported to be causing wood rot on apricot (2) and sweet and sour cherry (3). To our knowledge, this is the first report of H. rosae causing dry fruit rot on a stone fruit species in the world. References: (1) K. K. Nakasone. Mycologie, 29:231, 2008. (2) J. M. Ogawa et al. Diseases of Apricot (Prunus armeniaca L.). The American Phytopathological Society, St. Paul, MN, 2003. (3) J. K. Uyemoto et al. Diseases of Sweet Cherry (Prunus avium L.) and Sour Cherry (P. cerasus L.). IS-MPMInet, http://www.ismpminet.org/resources/common/comment/cherry.asp , accessed June 2012. (4) T. J. White et al. Page: 315 in: PCR Protocols: A Guide to Methods and Application. M.A. Innis et al., eds. Academic Press, San Diego, CA, 1990.

Identification of Helicobacter pylori DNA in Iranian patients with gallstones.

In order to identify Helicobacter in gallstones of Iranian patients with biliary disease, gallstone and bile samples from 33 patients were subjected to rapid urease test, culture and Multiplex PCR using primers based on 16s rRNA and isocitrate dehydrogenase genes for the identification of Helicobacter genus and H. pylori respectively. This PCR was also done on bile samples from 40 autopsied gallbladders with normal pathology (control group). In 18.1% of stone and 12.1% of bile samples, H. pylori DNA was detected using PCR. Rapid urease and culture tests were negative for all samples. The PCR was negative in the control group. In conclusion, H. pylori DNA was detected in stone samples of Iranian patients with gallstones but we are not sure of their viability. To clarify the clinical role of Helicobacter in gallbladder diseases, studies using accurate tests on larger patient and control groups are needed to ascertain whether this microorganism is an innocent bystander or active participant in gallstone formation.