Biochemical Monitoring of Spinal Cord Injury by FT-IR Spectroscopy—Effects of Therapeutic Alginate Implant in Rat Models

Spinal cord injury (SCI) induces complex biochemical changes, which result in inhibition of nervous tissue regeneration abilities. In this study, Fourier-transform infrared (FT-IR) spectroscopy was applied to assess the outcomes of implants made of a novel type of non-functionalized soft calcium alginate hydrogel in a rat model of spinal cord hemisection (n = 28). Using FT-IR spectroscopic imaging, we evaluated the stability of the implants and the effects on morphology and biochemistry of the injured tissue one and six months after injury. A semi-quantitative evaluation of the distribution of lipids and collagen showed that alginate significantly reduced injury-induced demyelination of the contralateral white matter and fibrotic scarring in the chronic state after SCI. The spectral information enabled to detect and localize the alginate hydrogel at the lesion site and proved its long-term persistence in vivo. These findings demonstrate a positive impact of alginate hydrogel on recovery after SCI and prove FT-IR spectroscopic imaging as alternative method to evaluate and optimize future SCI repair strategies.     

Chromosomal investigation of three Costa Rican frogs from the 30-chromosome radiation of Hyla with the description of a unique geographic variation in nucleolus organizer regions

A cytogenetic investigation of Hyla ebraccata Cope, H. microcephala Cope, and H. phlebodes Stejneger revealed that the karyotypes of these 30-chromosome Hyla are very conservative. With the exception of some structural rearrangements, only few differences in chromosomal morphology could be discerned. Based on our results, we hypothesize that the telomeric position of nucleolus organizer regions (NOR) on chromosome no. 10 may represent a derived condition in 30-chromosome Hyla. This cytotype was found only in the Caribbean population of H. ebraccata, Such within-species disparity has not been observed previously among amphibians. This phenomenon can most readily be explained by a translocation or insertion that rapidly drifted to high frequency in a small population.     

Chromosome banding in Amphibia

The chromosomes of the South American marsupial frogs Gastrotheca fissipes, G. ovifera, G. walkeri and Flectonotus pygmaeus were analyzed by means of conventional and various banding techniques. The karyotypes of G. ovifera and G. walkeri are characterized by highly differentiated XY/XX sex chromosomes. Whereas the X chromosomes and autosomes contain large amounts of constitutive heterochromatin, extremely little heterochromatin is located in the Y chromosomes. This is in contrast to all previously known amphibian Y chromosomes and the Y chromosomes of most other vertebrates. In the male meiosis of G. walkeri, the euchromatic segments of the heteromorphic XY chromosomes show the same pairing configuration as the autosomal bivalents. The karyotype of F. pygmaeus is remarkable for the unique presence of telocentric chromosomes and the high frequency of interstitially located chiasmata in the meiotic bivalents. The evolution of the karyotypes and sex chromosomes, the structure of the various classes of heterochromatin and the data obtained from meiotic analyses of the marsupial hylids are discussed.     

Detection of a CfoI polymorphism within exon 5 of the human neuronal nicotinic acetylcholine receptor α4 subunit gene (CHRNA4)

The gene coding for the 4 subunit of the neuronal nicotinic acetylcholine receptor (CHRNA4) has been recently mapped in the candidate region for benign familial neonatal convulsions (BFNC) on chromosome 20q13.2–q13.3. The region is only partially covered with polymorphic markers, and so far no PCR-based polymorphisms have been described in the critical region for BFNC. We now report the first polymorphic marker in the coding region of CHRNA4. The new marker, which is detected by PCR, will be useful for evaluation of the role of CHRNA4 as a candidate gene for BFNC. It will further enable the investigation of this important brain-specific gene in association studies with different types of epileptic diseases and other neurological disorders.     

A novel way to induce erythroid progenitor self renewal: cooperation of c-Kit with the erythropoietin receptor

Red blood cells are of vital importance for oxygen transport in vertebrates. Thus, their formation during development and homeostasis requires tight control of both progenitor proliferation and terminal red cell differentiation. Self renewal (i.e. long-term proliferation without differentiation) of committed erythroid progenitors has recently been shown to contribute to this regulation. Avian erythroid progenitors expressing the EGF receptor/c-ErbB (SCF/TGFalpha progenitors) can be induced to long-term proliferation by the c-ErbB ligand transforming growth factor alpha and the steroids estradiol and dexamethasone. These progenitors have not yet been described in mammals and their factor requirements are untypical for adult erythroid progenitors. Here we describe a second, distinct type of erythroid progenitor (EpoR progenitors) which can be established from freshly isolated bone marrow and is induced to self renew by ligands relevant for erythropoiesis, i.e. erythropoietin, stem cell factor, the ligand for c-Kit and the glucocorticoid receptor ligand dexamethasone. Limiting dilution cloning indicates that these EpoR progenitors are derived from normal BFU-E/CFU-E. For a detailed study, mEpoR progenitors were generated by retroviral expression of the murine Epo receptor in bone marrow erythroblasts. These progenitors carry out the normal erythroid differentiation program in recombinant differentiation factors only. We show that mEpoR progenitors are more mature than SCF/TGFalpha progenitors and also do no longer respond to transforming growth factor alpha and estradiol. In contrast they are now highly sensitive to low levels of thyroid hormone, facilitating their terminal maturation into erythrocytes.     

Chromosome banding in Amphibia

The mitotic and meiotic chromosomes and interphase nuclei of the South American tree-frog Centrolenella antisthenesi were studied with various banding techniques. The karyotype is distinguished by a new category of heteromorphic XY/XX sex chromosomes in an initial stage of differentiation. In diakinesis of male meiosis the XY chromosomes exhibit the same pairing configuration as the autosomal bivalents. Analysis of the chromosomes with DNA base pair-specific fluorochromes revealed that unusual large amounts of brightly labeled AT-rich constitutive heterochromatin are located in the centromeric and pericentromeric regions of all autosomes and in the X chromosome. In most types of interphase cell nuclei the brightly fluorescent heterochromatic regions fuse to very large chromocenters.     

A mid-Holocene vegetational and anthropogenic record in the Guanabara Bay region,Rio de Janeiro State,SE Brazil,assessed by palynological and charcoal data

Palynological and charcoal fragment analyses of Guanabara Bay sediments, as well as radiocarbon dating, were carried out on one 220 cm long sedimentary core collected from the northeast sector of the bay, near the São Gonçalo coast. This study aims at recognising and explaining the environmental history of this region during the mid-Holocene, and to identify the anthropogenic influences on it. The palynological data indicate the predominance of ombrophilous forest vegetation in the Guanabara Bay Hydrographic Basin at c. 6500 calendar years bp. During this period (pollen zone 1), the concentrations of pollen grains and spores may still have been controlled by the Holocene Maximum Transgressive Event. After a phase of low concentrations of palynomorphs, there was a considerable increase in the accumulation patterns of pollen and spore grains (pollen zone 2). This increase can probably be related to the retreat of the sea level event after the Holocene Maximum Transgressive Event. The 75 cm of the core top (pollen zone 3) provide clear evidence of human influence in the area, which is inferred from the significant reduction in ombrophilous forest pollen grains, the significant increase in herbaceous pollen grains, the presence of exotic pollen types (Eucalyptus and Pinus), and high concentration of carbonaceous particles.     

Cellular and extracellular miRNAs are blood‐compartment‐specific diagnostic targets in sepsis

Septic shock is a common medical condition with a mortality approaching 50% where early diagnosis and treatment are of particular importance for patient survival. Novel biomarkers that serve as prompt indicators of sepsis are urgently needed. High‐throughput technologies assessing circulating microRNAs represent an important tool for biomarker identification, but the blood‐compartment specificity of these miRNAs has not yet been investigated. We characterized miRNA profiles from serum exosomes, total serum and blood cells (leukocytes, erythrocytes, platelets) of sepsis patients by next‐generation sequencing and RT‐qPCR (n = 3 × 22) and established differences in miRNA expression between blood compartments. In silico analysis was used to identify compartment‐specific signalling functions of differentially regulated miRNAs in sepsis‐relevant pathways. In septic shock, a total of 77 and 103 miRNAs were down‐ and up‐regulated, respectively. A majority of these regulated miRNAs (14 in serum, 32 in exosomes and 73 in blood cells) had not been previously associated with sepsis. We found a distinctly compartment‐specific regulation of miRNAs between sepsis patients and healthy volunteers. Blood cellular miR‐199b‐5p was identified as a potential early indicator for sepsis and septic shock. miR‐125b‐5p and miR‐26b‐5p were uniquely regulated in exosomes and serum, respectively, while one miRNA (miR‐27b‐3p) was present in all three compartments. The expression of sepsis‐associated miRNAs is compartment‐specific. Exosome‐derived miRNAs contribute significant information regarding sepsis diagnosis and survival prediction and could serve as newly identified targets for the development of novel sepsis biomarkers.     

Spectral karyotyping of Werner syndrome fibroblast cultures

Fibroblast cultures from two Werner syndrome patients were analyzed by spectral karyotyping. There were multiple, pseudodiploid clones in both cultures, mostly marked by random balanced reciprocal translocations. One of the cultures contained a clone with three-way exchanges involving chromosomes 2, 3, and 16. Duplication-deficiencies were exceptional, as were completely normal metaphases. The most frequent breakpoint occurred at 16q22 which corresponds to FRA16B, possibly reflecting difficulties of WS cells in replicating AT-rich repetitive DNA structures. Both cultures ceased proliferation after eight in vitro passages, but a single clone with exceptional growth potential emerged in one of the senescing cultures. Due to its identical translocations, the derivation of this near tetraploid clone (with tetrasomy for all autosomes except chromosomes 4 and 6) could be traced to the most prevalent pseudodiploid clone of the parental mass culture. Our study confirms the existence of variegated translocation mosaicism as the cytogenetic hallmark of WS fibroblast cultures and suggests that tetraploidization in combination with certain chromosome rearrangements and selective chromosome dosage may overcome the severely limited in vitro lifespan of WS fibroblasts.