Estimates of DNA damage by the comet assay in the direct-developing frog Eleutherodactylus johnstonei (Anura, Eleutherodactylidae)

The aim of this study was to use the Comet assay to assess genetic damage in the direct-developing frog Eleutherodactylus johnstonei. A DNA diffusion assay was used to evaluate the effectiveness of alkaline, enzymatic and alkaline/enzymatic treatments for lysing E. johnstonei blood cells and to determine the amount of DNA strand breakage associated with apoptosis and necrosis. Cell sensitivity to the mutagens bleomycin (BLM) and 4-nitro-quinoline-1-oxide (4NQO) was also assessed using the Comet assay, as was the assay reproducibility. Alkaline treatment did not lyse the cytoplasmic and nuclear membranes of E. johnstonei blood cells, whereas enzymatic digestion with proteinase K (40 μg/mL) yielded naked nuclei. The contribution of apoptosis and necrosis (assessed by the DNA diffusion assay) to DNA damage was estimated to range from 0% to 8%. BLM and 4NQO induced DNA damage in E. johnstonei blood cells at different concentrations and exposure times. Dose-effect curves with both mutagens were highly reproducible and showed consistently low coefficients of variation (CV ≤ 10%). The results are discussed with regard to the potential use of the modified Comet assay for assessing the exposure of E. johnstonei to herbicides in ecotoxicological studies

Cruise Summary Report - MEDWAVES survey. MEDiterranean out flow WAter and Vulnerable EcosystemS (MEDWAVES)

The MEDWAVES (MEDiterranean out flow WAter and Vulnerable EcosystemS) cruise targeted areas under the potential influence of the MOW within the Mediterranean and Atlantic realms. These include seamounts where Cold-water corals (CWCs) have been reported but that are still poorly known, and which may act as essential “stepping stones” connecting fauna of seamounts in the Mediterranean with those of the continental shelf of Portugal, the Azores and the Mid-Atlantic Ridge. During MEDWAVES sampling has been conducted in two of the case studies of ATLAS: Case study 7 (Gulf of Cádiz-Strait of Gibraltar-Alboran Sea) and Case study 8 (Azores). The initially targeted areas in the Atlantic were: the Gazul Mud volcano, in the Gulf of Cádiz (GoC) area, included in the case study 7, and the Atlantic seamounts Ormonde (Portuguese shelf) and Formigas (by Azores), both part of the case study 8. In the Mediterranean the targeted areas were The Guadiaro submarine canyon and the Seco de los Olivos (also known as Chella Bank) seamount. Unfortunately it was not possible to sample in Guadiaro due to time constraints originated by adverse meteorological conditions which obligate us to reduce the time at sea focusing only in 4 of the 5 initially planned areas. MEDWAVES was structured in two legs; the first leg took place from the 21st September (departure from Cádiz harbour in Spain) to the 13th October 2016 (arrival in Ponta Delgada, São Miguel, Azores, Portugal took place the 8th of October due to the meteorological conditions that obligated to conclude the first leg earlier as planned). during the Leg 1 sampling was carried out in Gazul, Ormonde and Formigas. The second leg started the 14th October (departure from Ponta Delgada) and finished the 26th October (arrival in Málaga harbour, Spain). MEDWAVES had a total of 30 effective sampling days, being 6 days not operative due to the adverse meteorological conditions experienced during the first leg which forced us to stay in Ponta Delgada from the 08th to the 13th October. During MEDWAVES the daily routine followed a similar scheme, depending of course on the weather and sea conditions. The main activity during the day, starting early in the morning (around 08:00 AM, once the night activities were finished), was the ROV deployment. Generally a single ROV dive of around 8 hours was performed, however in several occasions two dives were carried out in the same day (see General station list, Appendix II). After the ROV (and sometimes between two dives) the Box Corer and/or Van Veen Grab and/or Multicore was deployed. After these activities, during the night CTD-Rosette deployments and MB was conducted. Accordingly to this schema the scientific personnel worked in the day or in the night watch. A total of 215 sampling stations have been covered in MEDWAVES, using the following sampling gears: Multibeam echosounder, CTD-Rosette, LADCP, Box Corer, Van Veen Grab, Multicorer and a Remotely Operated Vehicle (ROV). Table 1 sumamrised the number of sampling stations conducted with each gear in each sampling zone. Additionally MB surveys have been conducted during the transits between area

POLA KONSUMSI PANGAN GOITROGENIK DAN HUBUNGANNYA DENGAN UEI (KADAR IODIUM URIN) DI KABUPATEN CILACAP JAWA TENGAH

Goitrogenic essence can be easy to be find on the 'cheaper' foods, oc. on cabbages, mustard, leaves or roots of cassava and corn bean. (Matovinovic Josip, 1988). Increases of goitrogen food intake have more risk to be IDDI, especially on the low income and poor community. One of the measurement to know the risk level of IDDI by UEI (Urinary Excretion Iodine) examination.(Stanbury VB & Pinchera A, 1996). The one risk of IDDI is the low level of UEI on the fertility women. Indonesia Health Ministry (1996) appraises that UEI level on fertility women is important indicator to find out endemicity area for IDDI. The general aim of the study was to know about the relation between gotrogenic food intake and UEI level on the fertility women. The specific aims were to know (1) the kind of goitrogenic foods that usually be intaken by fertility women, (2) the pleasure level of each goitrogenic food and the average intake, (3) relation between income and goitrogenic food intake, (4) the UEI level and the endemicity of area, (5) relation between gotrogenic food intake and the UEI level, and (6) relation between each gotrogenic food and the UEI level. This survey was cross sectional that studied on fertility women at Cilacap District. Villages had been choised by purposive, appropriate to endemic area of Inn' Central Java Mapping (1996). 66 samples had been found by inclusion and exclusion criterias. The instrumens were (1) questionaire, (2) recall draft, (3) goitrogenic food draft, (4) indepth protocol, (5) wine bottle and (6) Iodina test. The data had been processed by PC¬SPSS-6 and analysed by Kendal Tau (non parametric test). The average of respondent age are 27,2 + 5,3 yo. The average of Education are 7,5 + 2,6 y. Almost of respondent are household mother (81,1 %), others are fanner, fanning laborer, private worker, government worker and trader. The average of monthly capita income are Rp 87.530,- ± Rp 53.490,-. Almost of salt at home have labelled iodium (98,5 %) and just one that haven't iodium labelled (1,5 %). Almost of salt have iodium level in accordance with standardization (63,6 %), 25,8 % not standardize and 0,6 % negative. Ten goitrogenic foods that been usually consumed by respondent are cassava leaves, cabbages, eggplant, papaya leaves soy sauce,stringbean, shallot, mustard greens, onion and tamarind. Four goitrogenic foods that most likely and largest consumed are cassava leaves, cabbages, eggplant and papaya leaves. The average of caloric daily gotrogenic foods (the largest) is cassava leaves (20,99 ± 17,16 cal). Maximal intake is cassava leaves too, until 75 cal/ day. Goitrogenic foods that were rarely consumed are 'petai' and lannoro' (leucaena glauca). If counted up the calories of gotrogenic foods, the minimally amount of caloric are 8,30 cal/day and maximally are 200,4 cal/day. The average are 74,28 + 48,87 cal/day. The average of UEI level are 350,45 ± 155,96 ug/L. Minimally UEI level are 44 ug/L dan maximally UEI level are 705 ug/L. We can find that average of UEI level more than 100 ug/L so the area aren t included the IDDI problem or that villages are not IDDI area anymore. The result of Kendal Tau test were (1) no relation between capita income and amount of gotrogenic consumtion (p = 0,731, t= - 0,0294); (2) no relation between amount of goitrogenic consumtion and UEI level (p = 0,816, t= - 0,0197); (3) if tested each of goitrogenic foods, there were three foods that had significant relations: petal, jengkol and eggplant. Negative relations could be found on petal and jengkol, but possitive relation on eggplant. It's mean that the lower UEI level could be found on the higher petal and jengkol intake, but the other way the lower UEI level could be found on the lower eggplant intake too. It's need to continue this study about the polite of eggplant on goitrogenic food classification and the advance study for petal and jengkol Zat goitrogenik tersebut banyak terdapat dalam bahan-bahan makanan yang relatif murah dan mudah didapat, antara lain: kobis (kol), sawi, daun dan akar ubi/ ketela pohon serta kacang kedelai. (Matovinovic Josip, 1988). Tingginya konsumsi zat goitrogen tersebut akan memperbesar peluang terjadinya GAKI pada masyarakat dengan penghasilan rendah dan tidak mampu. Sedangkan cam untuk mengetahui tingkat kerawanan GAKI di suatu daerah ada bermacam-macam diantaranya adalah dengan mengukur kadar iodium dalam urin (Urinary Excretion Iodine/ UEI).(Stanbury VB & Pinchera A, 1996). Salah sam risiko rawan GAKI adalah rendahnya nilai UEI terutama pada kelompok-kelompok rentan di masyarakat terutama pada Wanita Usia Subur (WUS). Depkes RI (1996) menilai bahwa pengukuran UEI pada WUS merupalcan indikator yang cukup panting untuk melihat apakah di daerab tersebut menmunyai kecenderungan menjadi daerah endemik atau tidak. Tujuan utnum dad penelitian ini adalah untuk mengetahui hubungan antara intake atau konsumsi pangan goitrogenik dengan kadar UEI pada WUS. Sedangkan tujuan khusus: (1) mengetahui jenis-jenis bahan pangan goitrogenik yang biasa dikonsumsi oleh masyarakat terutama wanita usia suburnya, (2) mengetahui tingkat kesukaan wanita usia subur terhadap masing-masing bahan pangan goitrogenik dan rata-rata tingkat konsumsinya, (3) mengetahui hubungan antara tingkat pendapatan dengan konsumsi pangan gotrogenik, (4) mengetahui kadar UEI pada wanita usia subur dan tingkat endemisitasnya, (5) mengetahui hubungan antara konsumsi pangan goitrogenik dengan kadar U1E dan (6) mengetahui hubungan masing-masing balm pangan gotrogenik terhadap kadar DEL Panetitian ini mempakan penelitian survey jenis crosssectional. Populasinya adalah wanita usia subur pada sebuah kecamatan di Kabupaten Cilacap yang topografinya pegunungan dan merupakan daerah endemis. Selanjutnya dad saw kecamatan tersebut dipilih saw desa secara purposive, yaitu desa yang pemah dipakai pada saat pemetaan GAKI tahun 1996. Selanjutnya dad desa tersebut diambil sampel minimal 30 wanita usia subur (bisa secara purposif, dengan kriteria inklusi: usia 20 — 35 tahun, sehat jasmani dan rokhani, menikah; sedangkan kriteria eksklusinya: hamil, menyusui serta adanya riwayat penyakit kronis. Instrumen yang dipergunakan adalah (1) kuesioner untuk data primer (2) Draft recall dua hart (3) Draft zat gotrogenik. (4) Kuesioner mendalmn tentang tingkat kesukaan terhadap pangan gotrogenik (5) Botol thin untuk pengambilan urin sewaktu (pemeriksaan UEI) dan (6) Iodina test untuk screening. Data diolah dengan PC-SPSS for Window versi 6, kemudian dianalisa secara deskriptif (univariat dan bivariat) serta secara inferensial (uji hubungan). Uji hubungan dengan uji korelasi kendal tau, karma didapatkan pada uniumnya data berdistribusi tidak normal dan diputuskan untuk memakai uji non parametrik. Umur rata-rata responden adalah 27,2 + 5,3 th. Pendidikan rata-rata 7,5 + 2,6 di. Sebagian besar responden tidak bekerja (81,1 %). Sebagian lagi sebagai petani pemilik, buruh tani, pegawai swasta, pegawai negeri dan berdagang. Pendapatan perkapita perbulan rata-rata adalah Rp 87.530,- + Rp 53.490,- .Garam yang ada di rumah hampir seluruhnya sudah berlabel iodium (98,5 %) dan hattya ada satu yang tidak berlabel iodium (1,5 %). Sebagian besar (63,6 %) garam mengandung kadar iodium yang sesuai. 25,8 % kadar iodium garam tidak sesuai dan 10,6 % tidak mengandung iodium sama sekali (negatif). Sepuluh bahan pangan yang biasa dikonsumsi oleh responden adalah daun singkong, kobis, terung, dam pepaya, kecap, buncis, brambang, saws, bawang dan asam jawa. Sedangkan empat macam bahan pangan dari yang paling disukai dan banyak dikonsumsi adalah daun singkong, kobis, terung dan dam pepaya. Konsumsi pangan goitrogenik setelah dikonversikan dalam rata-rata kalori perhari, bahan pangan goitrogenik yang paling banyak mereka mengkonsumsi adalah dam singkong (20,99 ± 17,16 kal). Sedangkan jumlah maksimal yang dimakan juga daun singkong yaitu sampai 75 kalori perhari. Bahan pangan goitrogenik yang paling jarang dimalcan adalah petal dan lamtoro. Apabila kalori dari masing-masing bahan pangan goitrogenik dijumlahkan maka jumalh minimal adalah 8,30 kalori, maksimal 200,4 kalori perhari. Rata-rata bahan pangan goitrogenik yang dikonsumsi adalah 74,28 + 48,87 kalori. Kadar UEI rata-rata adalah 350,45 + 155,96 ug/L. UEI minimal adalah 44 ug/L dan UEI maksimal 705 ug/L. Melihat kadar UEI rata-rata yang tinggi dengan nilai median lebih dari 100 ug/L maka daerah tersebut dapat dikatakan sudah tidak bennasalah GAM. Pada uji satistik didapatkan hasil sbb (1) tidak didapatkan adanya hubungan antara pendapatan perkapita dengan jumlah konsumsi pangan goitrogenik (p = 0,731, - 0,0294); (2) Tidak didapatkan adanya hubungan antara jumlah konsumsi pangan goitrogenik dengan kadar UEI (p = 0,816, t= - 0,0197); (3) dari kelimabelas balm' pangan goitrogenik, ada tiga yang ada hubungan signifikan yaitu petai, jengkol dan terung. Hubungan negatif terdapat pada petai dan jengkol artinya kadar UEI rendah terdapat pada konsumsi petai maupun jengkol yang tinggi, sedangkan hubungan positif pada terung yang berarti kadar UEI rendah terdapat pada konsumsi terung yang rendah pula. Perlu dikaji lebih lanjut keberadaan terung sebagai bahan pangan goitrogenik dan diharapkan ada penelitian yang !eta mendalam tentang petal dan jengkol. perbulan rata-rata adalah Rp 87.530,- + Rp 53.490,- .Garam yang ada di rumah hampir seluruhnya sudah berlabel iodium (98,5 %) dan hattya ada satu yang tidak berlabel iodium (1,5 %). Sebagian besar (63,6 %) garam mengandung kadar iodium yang sesuai. 25,8 % kadar iodium garam tidak sesuai dan 10,6 % tidak mengandung iodium sama sekali (negatif). Sepuluh bahan pangan yang biasa dikonsumsi oleh responden adalah daun singkong, kobis, terung, dam pepaya, kecap, buncis, brambang, saws, bawang dan asam jawa. Sedangkan empat macam bahan pangan dari yang paling disukai dan banyak dikonsumsi adalah daun singkong, kobis, terung dan dam pepaya. Konsumsi pangan goitrogenik setelah dikonversikan dalam rata-rata kalori perhari, bahan pangan goitrogenik yang paling banyak mereka mengkonsumsi adalah dam singkong (20,99 ± 17,16 kal). Sedangkan jumlah maksimal yang dimakan juga daun singkong yaitu sampai 75 kalori perhari. Bahan pangan goitrogenik yang paling jarang dimalcan adalah petal dan lamtoro. Apabila kalori dari masing-masing bahan pangan goitrogenik dijumlahkan maka jumalh minimal adalah 8,30 kalori, maksimal 200,4 kalori perhari. Rata-rata bahan pangan goitrogenik yang dikonsumsi adalah 74,28 + 48,87 kalori. Kadar UEI rata-rata adalah 350,45 + 155,96 ug/L. UEI minimal adalah 44 ug/L dan UEI maksimal 705 ug/L. Melihat kadar UEI rata-rata yang tinggi dengan nilai median lebih dari 100 ug/L maka daerah tersebut dapat dikatakan sudah tidak bennasalah GAM. Pada uji satistik didapatkan hasil sbb (1) tidak didapatkan adanya hubungan antara pendapatan perkapita dengan jumlah konsumsi pangan goitrogenik (p = 0,731, - 0,0294); (2) Tidak didapatkan adanya hubungan antara jumlah konsumsi pangan goitrogenik dengan kadar UEI (p = 0,816, t= - 0,0197); (3) dari kelimabelas balm' pangan goitrogenik, ada tiga yang ada hubungan signifikan yaitu petai, jengkol dan terung. Hubungan negatif terdapat pada petai dan jengkol artinya kadar UEI rendah terdapat pada konsumsi petai maupun jengkol yang tinggi, sedangkan hubungan positif pada terung yang berarti kadar UEI rendah terdapat pada konsumsi terung yang rendah pula. Perlu dikaji lebih lanjut keberadaan terung sebagai bahan pangan goitrogenik dan diharapkan ada penelitian yang !eta mendalam tentang petal dan jengkol

Variants in PRKAR1B cause a neurodevelopmental disorder with autism spectrum disorder, apraxia, and insensitivity to pain

Purpose We characterize the clinical and molecular phenotypes of six unrelated individuals with intellectual disability and autism spectrum disorder who carry heterozygous missense variants of the PRKAR1B gene, which encodes the R1 beta subunit of the cyclic AMP-dependent protein kinase A (PKA). Methods Variants of PRKAR1B were identified by single- or trio-exome analysis. We contacted the families and physicians of the six individuals to collect phenotypic information, performed in vitro analyses of the identified PRKAR1B-variants, and investigated PRKAR1B expression during embryonic development. Results Recent studies of large patient cohorts with neurodevelopmental disorders found significant enrichment of de novo missense variants in PRKAR1B. In our cohort, de novo origin of the PRKAR1B variants could be confirmed in five of six individuals, and four carried the same heterozygous de novo variant c.1003C>T (p.Arg335Trp; NM_001164760). Global developmental delay, autism spectrum disorder, and apraxia/dyspraxia have been reported in all six, and reduced pain sensitivity was found in three individuals carrying the c.1003C>T variant. PRKAR1B expression in the brain was demonstrated during human embryonal development. Additionally, in vitro analyses revealed altered basal PKA activity in cells transfected with variant-harboring PRKAR1B expression constructs. Conclusion Our study provides strong evidence for a PRKAR1B-related neurodevelopmental disorder

Heterozygous loss-of-function variants significantly expand the phenotypes associated with loss of GDF11

Growth differentiation factor 11 (GDF11) is a key signaling protein required for proper development of many organ systems. Only one prior study has associated an inherited GDF11 variant with a dominant human disease in a family with variable craniofacial and vertebral abnormalities. Here, we expand the phenotypic spectrum associated with GDF11 variants and document the nature of the variants.We present a cohort of six probands with de novo and inherited nonsense/frameshift (4/6 patients) and missense (2/6) variants in GDF11. We generated gdf11 mutant zebrafish to model loss of gdf11 phenotypes and used an overexpression screen in Drosophila to test variant functionality.Patients with variants in GDF11 presented with craniofacial (5/6), vertebral (5/6), neurological (6/6), visual (4/6), cardiac (3/6), auditory (3/6), and connective tissue abnormalities (3/6). gdf11 mutant zebrafish show craniofacial abnormalities and body segmentation defects that match some patient phenotypes. Expression of the patients’ variants in the fly showed that one nonsense variant in GDF11 is a severe loss-of-function (LOF) allele whereas the missense variants in our cohort are partial LOF variants.GDF11 is needed for human development, particularly neuronal development, and LOF GDF11 alleles can affect the development of numerous organs and tissues

Correction to: An autosomal dominant neurological disorder caused by de novo variants in FAR1 resulting in uncontrolled synthesis of ether lipids (Genetics in Medicine, (2021), 23, 4, (740-750), 10.1038/s41436-020-01027-3)

In the original author list, Seth Perlman’s degrees were listed as MD, PhD. Dr Perlman’s degree is MD. The original version has been corrected

An autosomal dominant neurological disorder caused by de novo variants in FAR1 resulting in uncontrolled synthesis of ether lipids

Purpose: In this study we investigate the disease etiology in 12 patients with de novo variants in FAR1 all resulting in an amino acid change at position 480 (p.Arg480Cys/His/Leu). Methods: Following next-generation sequencing and clinical phenotyping, functional characterization was performed in patients’ fibroblasts using FAR1 enzyme analysis, FAR1 immunoblotting/immunofluorescence, and lipidomics. Results: All patients had spastic paraparesis and bilateral congenital/juvenile cataracts, in most combined with speech and gross motor developmental delay and truncal hypotonia. FAR1 deficiency caused by biallelic variants results in defective ether lipid synthesis and plasmalogen deficiency. In contrast, patients’ fibroblasts with the de novo FAR1 variants showed elevated plasmalogen levels. Further functional studies in fibroblasts showed that these variants cause a disruption of the plasmalogen-dependent feedback regulation of FAR1 protein levels leading to uncontrolled ether lipid production. Conclusion: Heterozygous de novo variants affecting the Arg480 residue of FAR1 lead to an autosomal dominant disorder with a different disease mechanism than that of recessive FAR1 deficiency and a diametrically opposed biochemical phenotype. Our findings show that for patients with spastic paraparesis and bilateral cataracts, FAR1 should be considered as a candidate gene and added to gene panels for hereditary spastic paraplegia, cerebral palsy, and juvenile cataracts

Phenotypic expansion of CACNA1C-associated disorders to include isolated neurological manifestations

International audiencePurpose:CACNA1C encodes the alpha-1-subunit of a voltage-dependent L-type calcium channel expressed in human heart and brain. Heterozygous variants in CACNA1C have previously been reported in association with Timothy syndrome and long QT syndrome. Several case reports have suggested that CACNA1C variation may also be associated with a primarily neurological phenotype.Methods:We describe 25 individuals from 22 families with heterozygous variants in CACNA1C, who present with predominantly neurological manifestations.Results:Fourteen individuals have de novo, nontruncating variants and present variably with developmental delays, intellectual disability, autism, hypotonia, ataxia, and epilepsy. Functional studies of a subgroup of missense variants via patch clamp experiments demonstrated differential effects on channel function in vitro, including loss of function (p.Leu1408Val), neutral effect (p.Leu614Arg), and gain of function (p.Leu657Phe, p.Leu614Pro). The remaining 11 individuals from eight families have truncating variants in CACNA1C. The majority of these individuals have expressive language deficits, and half have autism.Conclusion:We expand the phenotype associated with CACNA1C variants to include neurodevelopmental abnormalities and epilepsy, in the absence of classic features of Timothy syndrome or long QT syndrome