Structure and Development of the Odontodes in an Armoured Catfish,Corydoras aeneus (Siluriformes,Callichthyidae)

Abstract In some living osteichthyans (e.g. the armoured catfishes) the postcranial dermal skeleton exhibits tooth-like structures (odontodes) similar to those present in the dermal skeleton of the ancient craniates. We have undertaken this work to compare odontode with tooth development, structure, attachment to a bony support and replacement. We studied the odontodes fixed on the scutes (i.e. postcranial dermal plates) in a growth series of Corydoras aeneus using light, scanning and transmission electron microscopy. Odontodes are constituted of a pulp cavity surrounded by a cone of dentine itself capped with hypermineralized substance. The pulp cavity is devoid of nerves and blood vessels and there are no odontoblastic processes in the dentine. The dentine cone is firmly attached to a circular bony protuberance of the scute surface, the pedicel or attachment bone, by means of a ligament. An odontode anlage develops as a small invagination of a dermal papilla projecting into the epidermis, the basal cell layer of which constitutes a dental epithelium. First, dentine is deposited, next the hypermineralized substance, then the ligament and attachment bone. Odontodes develop in two positions with regard to the scute surface: a primary position when new odontodes form at the posterior border of the enlarging scute; a secondary position when new odontodes replace old odontodes that have been shed during thickening of the scute. In this case, the ligament and part of the base of the dentine cone are resorbed but not the pedicel of attachment bone, which is covered by deposition of scute matrix after the odontode has been shed. Within the scute matrix, the embedded pedicels of successive generations of odontodes are preserved, forming piles in the scutes of adult specimens.     

SUMOylation mediates CtIP’s functions in DNA end resection and replication fork protection

Double-strand breaks and stalled replication forks are a significant threat to genomic stability that can lead to chromosomal rearrangements or cell death. The protein CtIP promotes DNA end resection, an early step in homologous recombination repair, and has been found to protect perturbed forks from excessive nucleolytic degradation. However, it remains unknown how CtIP’s function in fork protection is regulated. Here, we show that CtIP recruitment to sites of DNA damage and replication stress is impaired upon global inhibition of SUMOylation. We demonstrate that CtIP is a target for modification by SUMO-2 and that this occurs constitutively during S phase. The modification is dependent on the activities of cyclin-dependent kinases and the PI-3-kinase-related kinase ATR on CtIP’s carboxyl-terminal region, an interaction with the replication factor PCNA, and the E3 SUMO ligase PIAS4. We also identify residue K578 as a key residue that contributes to CtIP SUMOylation. Functionally, a CtIP mutant where K578 is substituted with a non-SUMOylatable arginine residue is defective in promoting DNA end resection, homologous recombination, and in protecting stalled replication forks from excessive nucleolytic degradation. Our results shed further light on the tightly coordinated regulation of CtIP by SUMOylation in the maintenance of genome stability.     

Response to Zeuthen and Zeuthen's Comment to the Editor: Enough Local Hypertonicity Is Enough

In a Comment to the Editor, Zeuthen and Zeuthen criticize our treatment of the water cotransport hypothesis. In this response, we argue that we calculated water cotransport as if there were no significant local osmotic gradient generated in the first minute of Na/glucose cotransport. It is surprising to receive this type of criticism from Zeuthen and Zeuthen, as the same treatment was used in at least six studies from his laboratory where it is systematically assumed that “intracellular unstirred layers effects” are negligible. Zeuthen and Zeuthen also state that “the cotransport hypothesis predicts the measurements better than the osmotic hypothesis”. We present a quantitative comparison that challenges this contention. We would like to conclude by stating that our article was not about comparing different numerical models but about an experimental measurement of the local osmotic gradient generated after 20, 40, or 60 s of cotransport. Osmotic gradients were indeed detected, and were of appropriate amplitude to explain virtually all water transport observed.     

Impact of benzalkonium chloride,benzethonium chloride and chloroxylenol on bacterial antimicrobial resistance

This review examined 3655 articles on benzalkonium chloride (BKC), benzethonium chloride (BZT) and chloroxylenol (CHO) aiming to understand their impact on antimicrobial resistance. Following the application of inclusion/exclusion criteria, only 230 articles were retained for analysis; 212 concerned BKC, with only 18 for CHO and BZT. Seventy-eight percent of studies used MIC to measure BKC efficacy. Very few studies defined the term ‘resistance’ and 85% of studies defined ‘resistance’ as <10-fold increase (40% as low as 2-fold) in MIC. Only a few in vitro studies reported on formulated products and when they did, products performed better. In vitro studies looking at the impact of BKC exposure on bacterial resistance used either a stepwise training protocol or exposure to constant BKC concentrations. In these, BKC exposure resulted in elevated MIC or/and MBC, often associated with efflux, and at time, a change in antibiotic susceptibility profile. The clinical relevance of these findings was, however, neither reported nor addressed. Of note, several studies reported that bacterial strains with an elevated MIC or MBC remained susceptible to the in-use BKC concentration. BKC exposure was shown to reduce bacterial diversity in complex microbial microcosms, although the clinical significance of such a change has not been established. The impact of BKC exposure on the dissemination of resistant genes (notably efflux) remains speculative, although it manifests that clinical, veterinary and food isolates with elevated BKC MIC carried multiple efflux pump genes. The correlation between BKC usage and gene carriage, maintenance and dissemination has also not been established. The lack of clinical interpretation and significance in these studies does not allow to establish with certainty the role of BKC on AMR in practice. The limited literature and BZT and CHO do not allow to conclude that these will impact negatively on emerging bacterial resistance in practice.     

Une forme de cryptorchidie d’origine génétique: le syndrome de persistance des canaux de Müller

Persistent Müllerian duct syndrome (PMDS), a rare form of male peudohermaphrodism, is characterized by lack of regression of Müllerian derivatives. These patients are externally phenotypic males in whom the presence of a uterus and Fallopian tubes is discovered during surgical correction of cryptorchidism and/or inguinal hernia. Molecular studies, in a total of 76 PMDS families, were performed by automatic sequencing after amplification by polymerase chain reaction (PCR) of different parts of the gene. AMH, synthesized by Sertoli cells, is a member of the Transforming Growth Factor-β superfamily. The 560 amino-acid glycoprotein is formed by two 70 kDa monomers linked by disulfide bonds. This hormone is cleaved at a proteolytic site 109 amino acids upstream of the C-terminus, yielding the bioactive C-terminal domain and a N-terminus which is not itself bioactive, but which enhances the bioactivity of the C-terminus. The gene, composed of five exons, is located on chromosome 19 (band p13.3). AMH gene mutations are present on the whole length of the gene in 47% of PMDS families. Sixty-one per cent were homozygous due to a high proportion of patients from Arabic or Mediterranean countries, characterized by a high rate of consanguinity. The serum AMH level, assessed by a commercially available enzyme immunoassay technique (ELISA), is extremely low in the great majority of patients, even before puberty when AMH levels are normally high. AMH binds to two distinct membrane-bound receptors, both serine/threonine kinases. The type II AMH receptor (AMHR-II) binds to the ligand, and this complex recruits receptor type I, which acts as a signal transducer by activating specific cytoplasmic substrates, the Smad molecules. AMHR-II, coded by a 8 kbp gene on chromosome 12 (band q13), contains 11 exons. Exons 1–3 encode the extracellular domain, exon 4 encodes the transmembrane part and exons 5–11 encode the intracellular serine/threonine kinase domain. An AMHR-II mutation was detected in 38% of PMDS families, characterized by a normal AMH level for the patient’s age. A particular mutation, a deletion of 27 bp in exon 10, was present in 45% of families of this group. No mutation of either AMH or the AMHR-II gene could be detected in 11 PMDS families.     

Dispersal and adaptive radiation of Bidens (Compositae) across the remote archipelagoes of Polynesia

The genus Bidens (Compositae) comprises c. 230 species distributed across five continents, with the 41 Polynesian species displaying the greatest ecomorphological variation in the group. However, the genus has had a long and complicated taxonomic history, and its phylogenetic and biogeographic history are poorly understood. To resolve the evolutionary history of the Polynesian Bidens, 152 individuals representing 91 species were included in this study, including 39 of the 41 described species from Polynesia. Four chloroplast and two nuclear DNA markers were utilized to estimate phylogenetic relationships, divergence times, and biogeographic history. Bidens was found to be polyphyletic within Coreopsis, consistent with previous assessments. The Polynesian radiation was resolved as monophyletic, with the initial dispersal into the Pacific possibly from South America to either the Hawaiian or Marquesas Islands. From the Marquesas, Bidens dispersed to the Society Islands, and ultimately to the Austral Islands. The initial diversification of the crown group in the Pacific is estimated to have occurred ~1.63 mya (0.74–2.72, 95% HPD), making Polynesian Bidens among the youngest and most rapid plant diversification events documented in the Pacific. Our findings suggest that relatively rare long‐distance dispersal and founder‐event speciation, coupled with subsequent loss of dispersal potential and within‐island speciation, can explain the repeated and explosive adaptive radiation of Bidens throughout the archipelagoes of Polynesia.     

Spring Time and Fall Time Effects of 1,25-Dihydroxyvitamin D3, Cobalt (II) and (CoCl2(1,25(OH)2D3)4) Complex on Renal and Cerebral Enzymatic Markers in Rats

The circadian activity rhythm undergoes changes in the course of postnatal development. Experiments without external time cues were performed to characterize the endogenous component and to investigate any age-dependent changes. Female laboratory mice were used. At the beginning of the experiment they were 3 (juvenile), 23 (adult) or 72 (senile) weeks old. Animals were kept in climatic chambers (constant darkness, food and water ad libitum, temperature: 22±2°C, rel. humidity: 55±5%). Locomotor activity was recorded continuously using infrared detectors. The data were stored and analysed by means of the “Chronobiology Kit” (Stanford University). The mean period lengths were not statistically different between age groups. The stability of the spontaneous activity rhythms was highest in adult mice, however. The mean activity/day decreased from juvenile to senile mice. A nonlinear interrelationship between period length and amount of activity was obtained. At lower activity levels the period length became shorter with increasing activity; at higher levels it became longer again. The general shape of the curve was similar in all age groups. With respect to the nonlinear curve, one could not establish a general age dependency of period length. At similar ranges of activity the period length would be shortest in senile animals. Taking into account, however, the decline with age of the amount of activity the period of old mice could be shorter than, equal to or longer than that of adult mice. The results show that the endogenous component of the circadian activity rhythm, including feedback loops, matures and stabilizes from the juvenile to the adult. An expected loss of stability in senile mice was not demonstrated, probably due to a high variance of the animals’ biological age. These age-dependent changes contribute to the changes of circadian activity rhythms obtained under entrained conditions.     

MRE11-RAD50-NBS1 Complex Is Sufficient to Promote Transcription by RNA Polymerase II at Double-Strand Breaks by Melting DNA Ends

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Identification of 96 single nucleotide polymorphisms in eight genes involved in iron metabolism: efficiency of bioinformatic extraction compared with a systematic sequencing approach

Single nucleotide polymorphisms (SNPs) can significantly contribute to the characterization of the genes predisposing to iron overloads or deficiencies. We report an SNP survey of coding and non-coding regions of eight genes involved in iron metabolism, by two successive methods. First, we made use of the public domain sequence data, by using assembled expressed sequence tags, non-redundant sequences, and SNP database screening. We extracted 77 potential SNPs of which only 31 could be further validated by sequencing DNA from 44 unrelated multi-ethnic individuals. Our results indicate that a bioinformatic approach may be effective only in those cases where candidate SNPs are extracted from two different data sources or in cases of experimentally confirmed SNPs. Second, additional systematic sequencing of DNA from 24 unrelated Breton subjects increased the number of SNPs over a total length of 86 kb to 96. The average distance between the SNPs and minor allele frequencies were higher than reported by others authors; this discrepancy may reflect the nature of the genes studied and the ethnic homogeneity of our test population.