Treatment with sorafenib in advanced thyroid cancer - a case report

Papillary thyroid cancer (PTC) usually has a good prognosis. The treatment, including total thyroidectomy and complementary radioiodine (RAI) therapy, gives complete remission in 90% of patients. However, in 10% of subjects with metastatic disease, the prognosis is poor. In the group of patients with disease progression and no 131I uptake, searching for new therapeutic modalities before all tyrosine kinase inhibitors and other antiangiogenic agents is necessary. The study presents the case of a 55-year-old male with advanced PTC /pT3mNxMo/ diagnosed in 1993. Primary treatment by total thyroidectomy and 131I ablation led to complete remission. In 2000 local as well as lymph node recurrence was diagnosed and successively treated by surgery. In 2006 an increasing serum thyroglobulin level was noted and a single lung metastasis was diagnosed and operated on. In 2007 new foci in CNS and vertebral column with no 131I uptake were stated. Further progression (bones, CNS, and pterygoid muscle) was confirmed by PET-CT. The patient underwent neurosurgical metastasectomy twice and palliative CNS and vertebra's radiotherapy. Liver metastases were diagnosed in 2009. Treatment with increasing doses of thalidomide (up to 800 mg/d) was administered for 3 months with a good tolerance; however, the therapy was withdrawn due to cancer progression. Next, sorafenib (800 mg/d) was given for 16 weeks. Radiological examination performed after 16 weeks confirmed stable disease, whereas 2 months later, after sorafenib withdrawal due to lack of treatment possibility, further progression was observed. Metronomic chemotherapy with Adriamycin was instituted which gave disease stabilization for 6 months. The patient died with advanced disseminated disease due to pulmonary embolism. We present this case to document no adverse effects of therapy with sorafenib in a patient with brain DTC metastases. Sorafenib therapy was only short-term, but no progression occurred in this time.     

The Way to a Man's Heart Is through His Stomach: What about Horses?

Background

How do we bond to one another? While in some species, like humans, physical contact plays a role in the process of attachment, it has been suggested that tactile contact''s value may greatly differ according to the species considered. Nevertheless, grooming is often considered as a pleasurable experience for domestic animals, even though scientific data is lacking. On another hand, food seems to be involved in the creation of most relationships in a variety of species.

Methodology/Principal Findings

In this study, we used the horse training context to test the effects of food versus grooming during repeated human-horse interactions. The results reveal that food certainly holds a key role in the attachment process, while tactile contact was here clearly insufficient for bonding to occur.

Conclusion/Significance

This study raises important questions on the way tactile contact is perceived, and shows that large inter-species differences are to be expected.     

Does the KIR2DS5 Gene Protect from Some Human Diseases?

Background

KIR2DS5 gene encodes an activating natural killer cell receptor whose ligand is not known. It was recently reported to affect the outcome of hematopoietic stem cell transplantation.

Methodology/Principal Findings

In our studies on KIR2DS5 gene associations with human diseases, we compared the frequencies of this gene in patients and relevant controls. Typing for KIR2DS5 gene was performed by either individual or multiplex polymerase chain reactions which, when compared in the same samples, gave concordant results. We noted an apparently protective effect of KIR2DS5 gene presence in several clinical conditions, but not in others. Namely, this effect was observed in ankylosing spondylitis (p = 0.003, odds ratio [OR] = 0.47, confidence interval [CI] = 0.28–0.79), endometriosis (p = 0.03, OR = 0.25, CI = 0.07–0.82) and acute rejection of kidney graft (p = 0.0056, OR = 0.44, CI = 0.24–0.80), but not in non-small-cell lung carcinoma, rheumatoid arthritis, spontaneous abortion, or leukemia (all p>0.05). In addition, the simultaneous presence of KIR2DS5 gene and HLA-C C1 allotype exhibited an even stronger protective effect on ankylosing spondylitis (p = 0.0003, OR = 0.35, CI = 0.19–0.65), whereas a lack of KIR2DS5 and the presence of the HLA-C C2 allotype was associated with ankylosing spondylitis (p = 0.0017, OR = 1.92, CI = 1.28–2.89), whereas a lack of KIR2DS5 and presence of C1 allotype was associated with rheumatoid arthritis (p = 0.005, OR = 1.47, CI = 1.13–1.92). The presence of both KIR2DS5 and C1 seemed to protect from acute kidney graft rejection (p = 0.017, OR = 0.47, CI = 0.25–0.89), whereas lack of KIR2DS5 and presence of C2 seemed to favor rejection (p = 0.0015, OR = 2.13, CI = 1.34–3.37).

Conclusions/Significance

Our results suggest that KIR2DS5 may protect from endometriosis, ankylosing spondylitis, and acute rejection of kidney graft.     

Whole-Genome Profiling of Mutagenesis in Caenorhabditis elegans

Deep sequencing offers an unprecedented view of an organism''s genome. We describe the spectrum of mutations induced by three commonly used mutagens: ethyl methanesulfonate (EMS), N-ethyl-N-nitrosourea (ENU), and ultraviolet trimethylpsoralen (UV/TMP) in the nematode Caenorhabditis elegans. Our analysis confirms the strong GC to AT transition bias of EMS. We found that ENU mainly produces A to T and T to A transversions, but also all possible transitions. We found no bias for any specific transition or transversion in the spectrum of UV/TMP-induced mutations. In 10 mutagenized strains we identified 2723 variants, of which 508 are expected to alter or disrupt gene function, including 21 nonsense mutations and 10 mutations predicted to affect mRNA splicing. This translates to an average of 50 informative mutations per strain. We also present evidence of genetic drift among laboratory wild-type strains derived from the Bristol N2 strain. We make several suggestions for best practice using massively parallel short read sequencing to ensure mutation detection.MUTAGENESIS and the screening for mutants have long been a key tool of the practicing geneticist. The early work of T. H. Morgan and his colleagues relied on recovery of spontaneous mutations, which was limiting for the study of inheritance due to their infrequent occurrence (Morganet al. 1922; also see Sturtevant 1965). The discovery by H. J. Muller and others that X rays cause mutations ushered in the era of inducing mutations (Muller 1927). There is a long history of studies on mutagen specificity, both in prokaryotes and in eukaryotes, and today many mutagens are utilized in a variety of model organisms. In this article we use whole-genome deep sequencing in the model organism Caenorhabditis elegans to explore the types and frequencies of mutations induced by various mutagens and to document the feasibility of global identification of mutations.The mutagenic properties of ethyl methanesulfonate (EMS) were first demonstrated using the T4 viral system (Loveless 1959). Soon after, Lewis and Bacher (1968) demonstrated how to administer EMS to Drosophila melanogaster to generate mutations, and later Sydney Brenner did the same for the nematode C. elegans (Brenner 1974). The now classic article by Coulondre and Miller (1977) demonstrated the types of nucleotide substitutions generated by EMS and confirmed earlier observations (Bautz and Freese 1960) concerning the strong bias for GC to AT transitions. Today, EMS is still the most powerful and popular mutagen used by researchers studying D. melanogaster and C. elegans. Purely on the basis of genetic inference, when used at a concentration of 50 mm, EMS is calculated to induce ∼20 function-affecting variant alleles in C. elegans strains derived using this mutagen (Greenwald and Horvitz 1982; Anderson 1995).The chemical N-ethyl-N-nitrosourea (ENU) has been used as a mutagen since the 1970s but came to prominence when it was demonstrated to be the most effective chemical mutagen in mice (Russell et al. 1979). Today it is still the chemical mutagen of choice for this organism (Anderson 2000; Acevedo-Arozena et al. 2008). ENU has also been used for C. elegans mutagenesis (De Stasio et al. 1997). Although it appears to have different biases with regard to gene targets and base changes relative to EMS, the background mutational load after ENU mutagenesis has not been fully characterized (De Stasio and Dorman 2001).The chemical 4,5′,8-trimethylpsoralen is a crosslinking agent that is activated by near ultraviolet light. Studies in Escherichia coli have shown that it causes both single-base changes and deletions (Piette et al. 1985; Sladek et al. 1989). C. elegans researchers became interested in the potential of ultraviolet trimethylpsoralen (UV/TMP) to generate deletions in worms after the first deletions in this organism were isolated using this mutagen (Yandell et al. 1994). UV/TMP is now a major reagent in the arsenal of the C. elegans knockout consortium laboratories (Barstead and Moerman 2006). As a tool for generating deletions in eukaryotes it is quite useful but, outside of studies on prokaryotes, little else is known about the spectrum of mutagenic effects caused by UV/TMP.Massively parallel short read sequencing technologies offer unprecedented opportunities to study the complete genetic complement of an individual organism (Hillier et al. 2008). For genetic model systems the impact of this technology extends to the identification and correlation of induced mutations with selected phenotypes (Sarin et al. 2008). Several of the technological and bioinformatic issues that arise with next generation sequencing have already been addressed for the nematode C. elegans (Hillier et al. 2008; Sarin et al. 2008; Shen et al. 2008; Rose et al. 2010). Still, it is not clear how deeply one must sequence to confidently identify a relevant variant allele in a target mutant strain. Also of importance are questions concerning mutagen choice and dosage as they relate to the rate of induction of new mutations and background mutational load. We have undertaken the following study on mutagenesis and mutation detection to establish the parameters necessary to exploit next generation sequencing technologies for C. elegans genetics. For the first time we offer a whole-genome direct measure of mutation spectrum and background load for EMS, ENU, and UV/TMP. Readers interested in whole-genome sequencing of EMS mutagenized strains in C. elegans should also see the accompanying article in this issue by Sarin et al. (2010). In our study we also measured the single-nucleotide variation among currently used wild-type strains. In addition, we measured sequence read depth of all sequence and coding sequence and from this we make a recommendation of average genome coverage to ensure the correct identification of the causative mutation. We also examined the issue of false positive and false negative calls and make recommendations to eliminate most false positives without losing bona fide mutations.     

Proteolytic events in cryonecrotic cell death: Proteolytic activation of endonuclease P23

Although cryosurgery is attaining increasing clinical acceptance, our understanding of the mechanisms of cryogenic cell destruction remains incomplete. While it is generally accepted that cryoinjured cells die by necrosis, the involvement of apoptosis was recently shown. Our studies of liver cell death by cryogenic temperature revealed the activation of endonuclease p23 and its de novo association with the nuclear matrix. This finding is strongly suggestive of a programmed-type of cell death process. The presumed order underlying cryonecrotic cell death is addressed here by examining the mechanism of p23 activation. To that end, nuclear proteins that were prepared from fresh liver, which is devoid of p23 activity, were incubated with protein fractions isolated from liver exposed to freezing/thawing that possessed a presumed p23 activation factor. We observed that the activation of p23 was the result of a proteolytic event in which cathepsin D played a major role. Different patterns of proteolytic cleavage of nuclear proteins after in vitro incubation of nuclei and in samples isolated from frozen/thawed liver were observed. Although both processes induced p23 activation, the incubation experiments generated proteolytic hallmarks of apoptosis, while freezing/thawing of whole liver resulted in typical necrotic PARP-1 cleavage products and intact lamin B. As an explanation we offer a hypothesis that after freezing, cells possess the potential to die through necrotic as well as apoptotic mechanisms, based on our finding that the cytosol of cells exposed to cryogenic temperatures contains both necrotic and apoptotic executors of cell death.     

Algal community patterns in Slovenian bogs along environmental gradients

In 2005 and 2006, epiphyton samples were collected from seven lowland and montane peat bogs in Slovenia. Water temperature, pH, conductivity, dissolved oxygen and saturation were measured at the same time. Diatoms, desmids and Cyanobacteria were the most abundand groups in species number. Canonical Correspondence Analysis (CCA) was carried out on Cyanobacteria, diatom and desmid flora composition. This analysis showed that shading was the most important parameter in Cyanobacteria distribution and bedrock the most important one in that of diatoms and desmids. Cluster analyses were carried out based on the Cyanobacteria, diatom and desmid data. The Cyanobacteria and diatom data separated sites, whereas the desmid data revealed a temporal aspect.