Mesenchymal chondrosarcoma - A rare cause of pleural effusion.

Mesenchymal chondrosarcoma is a malignant neoplasm arising from cartilaginous bone or soft tissue. It is uncommon yet devastating. Our patient was a 21-year-old man who presented with pleuritic chest pain and weight loss. His chest radiograph showed left pleural effusion. His pleural effusion analysis was consistent with exudative pleural effusion. Tuberculosis workup was negative. Pleural fluid cytology did not yield malignant cells. Subsequently, his computed tomography of thorax showed left rib sclerotic lesion with soft tissue component. Biopsy of the soft tissue eventually confirmed the diagnosis of mesenchymal chondrosarcoma. He succumbed to his illness before the diagnosis was confirmed. We hope that through this case report, we are able to provide some insight into this rare condition.

Kappa Opioid Receptors in Mesolimbic Terminals Mediate Escalation of Cocaine Consumption.

Increases in drug consumption over time, also known as escalation, is a key behavioral component of substance use disorder (SUD) that is related to potential harm to users, such as overdose. Studying escalation also allows researchers to investigate the transition from casual drug use to more SUD-like drug use. Understanding the neurobiological systems that drive this transition will inform therapeutic treatments in the aim to prevent increases in drug use and the development of SUD. The kappa opioid receptor (KOR) system is typically known for its role in negative affect, which is commonly found in SUD as well. Furthermore, the KOR system has also been implicated in drug use and importantly, modulating the negative effects of drug use. However, the specific neuronal subpopulation expressing KOR involved has not been identified. Here, we first demonstrated that pharmacologically inhibiting KOR in the nucleus accumbens core (NAcC), as a whole, blocks cocaine escalation under long-access self-administration conditions. We then demonstrated that KOR expressed on ventral tegmental area (VTA) neurons but not NAcC neurons is sufficient for blocking cocaine escalation by utilizing a novel virally-mediated CRISPR-SaCas9 knock-out of the oprk1 gene. Together, this suggests that activation of KOR on VTA terminals in the NAcC drives the transition to the SUD-like phenotype of escalation of cocaine consumption.

Molecular, serological and biological variation among chickpea chlorotic stunt virus isolates from five countries of North Africa and West Asia.

Chickpea chlorotic stunt virus (CpCSV), a proposed new member of the genus Polerovirus (family Luteoviridae), has been reported only from Ethiopia. In attempts to determine the geographical distribution and variability of CpCSV, a pair of degenerate primers derived from conserved domains of the luteovirus coat protein (CP) gene was used for RT-PCR analysis of various legume samples originating from five countries and containing unidentified luteoviruses. Sequencing of the amplicons provided evidence for the occurrence of CpCSV also in Egypt, Morocco, Sudan, and Syria. Phylogenetic analysis of the CP nucleotide sequences of 18 samples from the five countries revealed the existence of two geographic groups of CpCSV isolates differing in CP sequences by 8-10%. Group I included isolates from Ethiopia and Sudan, while group II comprised those from Egypt, Morocco and Syria. For distinguishing these two groups, a simple RFLP test using HindIII and/or PvuII for cleavage of CP-gene-derived PCR products was developed. In ELISA and immunoelectron microscopy, however, isolates from these two groups could not be distinguished with rabbit antisera raised against a group-I isolate from Ethiopia (CpCSV-Eth) and a group-II isolate from Syria (CpCSV-Sy). Since none of the ten monoclonal antibodies (MAbs) that had been produced earlier against CpCSV-Eth reacted with group-II isolates, further MAbs were produced. Of the seven MAbs raised against CpCSV-Sy, two reacted only with CpCSV-Sy and two others with both CpCSV-Sy and -Eth. This indicated that there are group I- and II-specific and common (species-specific) epitopes on the CpCSV CP and that the corresponding MAbs are suitable for specific detection and discrimination of CpCSV isolates. Moreover, CpCSV-Sy (group II) caused more severe stunting and yellowing in faba bean than CpCSV-Eth (group I). In conclusion, our data indicate the existence of a geographically associated variation in the molecular, serological and presumably biological properties of CpCSV.

Molecular identification, incidence and phylogenetic analysis of seven viruses infecting garlic in Ethiopia.

Little information exists on the type and incidence of viruses infecting garlic (Allium sativum L) in Ethiopia. Attempts were made to identify the viruses using molecular techniques from 95 composite leaf samples collected from 44 farmers' fields and 51 germplasm accessions. Reverse transcription (RT-) PCR using genus and/or virus specific primers was used to amplify partial genome sequences of potyviruses, allexiviruses, carlaviruses and a tospovirus followed by sequencing of PCR products. Results indicated that ~73.7% of the samples are infected with at least one virus. Onion yellow dwarf virus (OYDV, genus Potyvirus, family Potyviridae) is the most common virus detected followed by Garlic virus C (genus Allexivirus) and Shallot latent virus (SLV, genus Carlavirus). Other viruses detected at lower frequency include Garlic virus X and Garlic virus D (genus Allexivirus), Leek yellow stripe virus (genus Potyvirus) and Iris yellow spot virus (IYSV, genus Tospovirus). Mixed infection of two or more viruses was detected in 65.7% of the samples. Phylogenetic analysis suggested that the different viruses may have been introduced to Ethiopia from Europe or Asia. This is the first report of Garlic virus X, Garlic virus D, IYSV and SLV in garlic in Ethiopia. The high incidence of OYDV and IYSV which cause severe yield loss alone or in mixed infection with allexiviruses and carlaviruses is a cause of concern to growers.

Identification and molecular characterization of Taro bacilliform virus and Taro bacilliform CH virus from East Africa.

Taro (Colocasia esculenta) and tannia (Xanthosoma sp.) are important root crops cultivated mainly by small-scale farmers in sub-Saharan Africa and the South Pacific. Viruses are known to be one of the most important constraints to production, with infections resulting in severe yield reduction. In 2014 and 2015, surveys were conducted in Ethiopia, Kenya, Tanzania and Uganda to determine the identity of viruses infecting taro in East Africa. Screening of 392 samples collected from the region using degenerate badnavirus primers revealed an incidence of 58-74% among the four countries surveyed, with sequence analysis identifying both Taro bacilliform virus (TaBV) and Taro bacilliform CH virus (TaBCHV). TaBCHV was identified from all four countries while TaBV was identified in all except Ethiopia. Full-length sequences from representative TaBV and TaBCHV isolates showed that the genome organization of TaBV isolates from East Africa was consistent with previous reports while TaBCHV isolates from East Africa were found to encode only four ORFs, distinct from a previous report from China. Phylogenetic analysis showed that all East African TaBV isolates form a single subgroup within known TaBV isolates, while TaBCHV isolates form at least two distinct subgroups. To the authors' knowledge, this is the first report describing the occurrence and genome organization of TaBV and TaBCHV isolates from East Africa and the first full-length sequence of the two viruses from tannia.

First Report of Soybean dwarf virus (Genus Luteovirus) Infecting Faba Bean and Clover in Germany.

In 2003, leaf samples from faba bean plants (Vicia faba L.) showing slight growth reductions and yellowing symptoms were collected in a field near Hebenshausen, Hesse, Germany. Some of these samples did not react in triple-antibody sandwich enzyme-linked immunosorbent assay (TAS-ELISA) with species-specific monoclonal antibodies (Mabs) to either Bean leaf roll virus or Turnip yellows virus, but did react with a broad-spectrum Mab (B-2-5G4) used to detect viruses in the genera Polerovirus and Luteovirus (family Luteoviridae) (1). Since this indicated the occurrence of a hitherto unrecognized polero- or luteovirus in faba bean in Germany, attempts were made to obtain nucleotide sequence information on two of the unknown faba bean isolates using a pair of degenerate primers (S2 [5'-ATCACITTCGGGCCGWSTCTATCAGA-3'] and AS3 [5'-CACGCGTCIACCTATTTIGGRTTITG-3'] [I = inosine]) derived from conserved domains in the capsid protein (CP) genes of several polero- and luteoviruses. Following reverse transcription (RT)-PCR amplification and cloning, the CP gene sequences of two genetically distinct isolates of Soybean dwarf virus (SbDV), a species of the genus Luteovirus, were obtained. To our knowledge, SbDV has not been reported from Germany or Europe but only from Africa, Australia, Japan, and the United States. In the two latter countries, at least two SbDV strain groups, SbDV-Y (for yellowing) and SbDV-D (for dwarfing), are distinguished on the basis of differences in symptomatology, host range, and molecular properties (2-4). On the basis of CP aa sequences, the two faba bean isolates from Hebenshausen differed by 8%, with one (FB1) most similar (>96% identity) to SbDV-D isolates and the other (FB2) closely related (>96%) to SbDV-Y isolates. Similar to observations in Japan (3) and the United States (2), we were able to detect SbDV in numerous samples from red clover (Trifolium pratense) and white clover (T. repens) in Braunschweig using SbDV antibodies (Agdia, Elkhart, IN) in DAS-ELISA. This was confirmed by RT-PCR amplification of CP gene sequences using SbDV-specific primers (SbDVs: 5'-GTCTACCTAAAAATTTCAAAGAATCTG-3'; SbDVas: 5'-CGGACCCGGTTCTCCGTCTA-3'). CP sequence analysis of two SbDV-positive clover samples revealed the presence of a SbDV-D isolate in red clover. However, a white clover plant contained an unusual SbDV isolate that possessed a unique CP, sharing aa sequence identities of approximately 92% with the two faba bean isolates from Germany and only 88.5 to 90.5% with other SbDV isolates. Attempts at aphid transmission of SbDV isolates from clover to faba bean were only successful for the combination Acyrthosiphon pisum and the white clover isolate. No faba bean seedlings became infected when the aphid species Aulacorthum solani and Aphis craccivora were given acquisition access feedings of 48 to 72 h on SbDV-infected white and red clover plants. The sequences determined in this study were deposited in GenBank (Accession Nos. EF466131-EF466134). References: (1) A. D. Abraham et al. Phytopathology 96:437, 2006. (2) V. D. Damsteegt et al. Phytopathology 89:374, 1999. (3) T. Tamada and M. Kojima. No. 179 in: Descriptions of Plant Viruses. Assoc. Appl. Biol. Kew, England, 1977. (4) H. Terauchi et al. Arch. Virol. 146:1885, 2001.

Chickpea chlorotic stunt virus: A New Polerovirus Infecting Cool-Season Food Legumes in Ethiopia.

ABSTRACT Serological analysis of diseased chickpea and faba bean plantings with yellowing and stunting symptoms suggested the occurrence of an unknown or uncommon member of the family Luteoviridae in Ethiopia. Degenerate primers were used for reverse transcriptase-polymerase chain reaction amplification of the viral coat protein (CP) coding region from both chickpea and faba bean samples. Cloning and sequencing of the amplicons yielded nearly identical (96%) nucleotide sequences of a previously unrecognized species of the family Luteoviridae, with a CP amino acid sequence most closely related (identity of approximately 78%) to that of Groundnut rosette assistor virus. The complete genome (5,900 nts) of a faba bean isolate comprised six major open reading frames characteristic of polero-viruses. Of the four aphid species tested, only Aphis craccivora transmitted the virus in a persistent manner. The host range of the virus was confined to a few species of the family Fabaceae. A rabbit antiserum raised against virion preparations cross-reacted unexpectedly with Beet western yellows virus-like viruses. This necessitated the production of murine monoclonal antibodies which, in combination with the polyclonal antiserum, permitted both sensitive and specific detection of the virus in field samples by triple-antibody sandwich, enzyme-linked immunosorbent assay. Because of the characteristic field and greenhouse symptoms in chickpea, the name Chickpea chlorotic stunt virus is proposed for this new member of the genus Polerovirus (family Luteoviridae).

Topoisomerase IIα mediates TCF-dependent epithelial-mesenchymal transition in colon cancer.

Aberrant T-cell factor (TCF) transcription is implicated in the majority of colorectal cancers (CRCs). TCF transcription induces epithelial-mesenchymal transition (EMT), promoting a tumor-initiating cell (TIC) phenotype characterized by increased proliferation, multidrug resistance (MDR), invasion and metastasis. The data presented herein characterize topoisomerase IIα (TopoIIα) as a required component of TCF transcription promoting EMT. Using chromatin immunoprecipitation (ChIP) and protein co-immunoprecipitation (co-IP) studies, we show that TopoIIα forms protein-protein interactions with ß-catentin and TCF4 and interacts with Wnt response elements (WREs) and promoters of direct target genes of TCF transcription, including: MYC, vimentin, AXIN2 and LEF1. Moreover, both TopoIIα and TCF4 ChIP with the N-cadherin promoter, which is a new discovery indicating that TCF transcription may directly regulate N-cadherin expression. TopoIIα N-terminal ATP-competitive inhibitors, exemplified by the marine alkaloid neoamphimedine (neo), block TCF activity in vitro and in vivo. Neo effectively inhibits TopoIIα and TCF4 from binding WREs/promoter sites, whereas protein-protein interactions remain intact. Neo inhibition of TopoIIα-dependent TCF transcription also correlates with significant antitumor effects in vitro and in vivo, including the reversion of EMT, the loss of TIC-mediated clonogenic colony formation, and the loss of cell motility and invasion. Interestingly, non-ATP-competitive inhibitors of TopoIIα, etoposide and merbarone, were ineffective at preventing TopoIIα-dependent TCF transcription. Thus, we propose that TopoIIα participation in TCF transcription may convey a mechanism of MDR to conventional TopoIIα inhibitors. However, our results indicate that TopoIIα N-terminal ATP-binding sites remain conserved and available for drug targeting. This article defines a new strategy for targeted inhibition of TCF transcription that may lead to effective therapies for the treatment of CRC and potentially other Wnt-dependent cancers.

The effects of thymomodulin and thymolymphotropin on the stress response of neuroendocrine system by gamma-irradiated Wistar rat.

Thymomodulin and Thymolymphotropin, biologically active thymus derivative peptides exert recovery effects on the functionality of some membrane bound, mitochondrial and lysosomal enzymes (monoamine oxidase, ATPase, phosphatases, cytochrome oxidase, succinate oxidase) affected by gamma-irradiation. These drugs exert antistress effect by re-establishing the function of hypothalamus-pituitary-adrenal axis and that of lymphoid organs.

L-tyrosine: 2-oxoglutarate aminotransferase induction by hydrocortisone in the thymus of the white rat.

Hydrocortisone hemisuccinate within 4 hours after in vivo administration produced an increase in precursor incoporation into rat thymus RNA and proteins in the whole animal. From these results, together with information obtained from measurements of the tyrosine aminotransferase activity and the action of mitomycin C administered one hour before the injection of hydrocortisone, it can be concluded that the increase in tissue level of the enzyme, consequent to hydrocortisone treatment, results from an increased rate of biosynthesis of the enzyme, which participates in the catabolic processes of proteins in glucocorticoid sensitive thymus cells.

'3H-testosterone distribution and binding in rat thymus cells in vivo'.

Autoradiography and biochemical investigations showed that [3H]-testosterone where injected intraperitoneally into male white rats was incorporated rapidly into thymus lymphocytes. Thymic cortex contained more silver grains than medulla, and larger lymphocytes were more labelled than medium or small lymphocytes. Cytosol fraction of thymus cells labelled in vivo with [3H]-testosterone, contained the largest quantity of labelled hormone. A 4S cytosol fraction binds [3H]-testosterone. This could be separated by Sephadex chromatography or by linear sucrose gradient centrifugation. Nuclear extract contained also a small quantity of the labelled hormone.

Histoenzymological and biochemical changes in the liver of adult female rats and of youngs born from Madiol-treated pregnant rats.

The effect of a 30-day treatment with Madiol was studied on the activity of some enzymes, nucleic acids, protein and glycogen content of the liver of adult female rats and youngs born from mothers treated during pregnancy. Madiol caused a significant increase in SDH and Atp-ase activity, and decreased glycogen and acid phosphatase.

Morphological and enzymological changes of small intestine of adult female rats and youngs born from madiol-treated pregnant rats.

Some histological and histoenzymological changes of the small intestine of adult female Wistar rats treated with Madiol were studied. The steroid did not influence the tissular aspect, but obviously stimulatory effects on protein and enzyme reactions were induced. Similar Madiol-action on the small intestine of 21-day-old young rats was observed when the steroid was administered during pregnancy to their mothers, suggesting influences on the embryonic development.

Release and retention of labelled DNA in the thymus and spleen of irradiated mice.

The process of release and retention of labelled DNA in thymus and spleen of normal and irradiated (60Co) mice has been studied after administration of 3H-thymidine. The results indicate that the dividing fraction of lymphoid cells is more resistant to radiation than the fraction of nondividing lymphocytes. The time courses of the specific activities of DNA in thymus and spleen were different especially after irradiation with lethal doses. It is suggested that the process of depletion in the lymphoid series is probably similar for both thymus and spleen but, the different cellular composition of these organs led to apparently unrelated results.

Effect of cortisol on the release and retention of labelled DNA in the thymus and spleen of mice.

The lymphocytolytic effect of different doses of cortisol was studied in the thymus and spleen of mice previously injected with 3H-thymidine. The results indicate that in thymus the fraction of labelled cells was more resistant to cortisol than the unlabelled cell population. The release of DNA into the fraction of DNA soluble in 0.14 M NaC1 was delayed suggesting that cortisol controls indirectly the lymphocytolytic process. Up to 24 hours after administration of cortisol the loss of labelled spleen cells significantly exceeded the loss of unlabelled cells. The time course of the release of labelled DNA into the fraction of DNA soluble in 0.14 M NaC1 indicates that a fraction of labelled DNA was rapidly removed from the spleen after injection of cortisol.

Effects of irradiation on the release and retention of labelled DNA in the small intestine of mice.

Mice were whole-body irradiated with 300, 500 and 800 rd (60Co) one hour or three days after administration of 3H-thymidine. The time courses of the total and specific activity of DNA indicate that three days after exposure with 500 and 800 rd the mitotically active cells were selectively lost from the small intestine. The release of labelled DNA was determined in the fraction of DNA soluble in 0.14 N NaCl. The results suggest that the relatively small fraction of soluble DNA belongs to the mitotically active cells probably having died in the interphase.

Evidence for reproductive death of dividing cells in thymus and spleen following whole-body gamma-irradiation of mice.

Mice were given whole-body gamma-irradiation with 300 and 800 rad 1 hour or 1 day after the administration of 3H-thymidine. The time-course of the specific activity of DNA in thymus and spleen and the release of radioactivity into the fraction of DNA soluble in 0-14 N NaCl were determined for 28 hours after exposure. The results indicate that on average the loss of mitotically-active cells was delayed compared with the loss of a representative fraction of non-dividing cells. This suggested that different mechanisms might be implicated in the dying process of the dividing and non-dividing lymphoid cells. Using a dose-fractionation method, the total activity of DNA was determined in thymus and spleen at 24 hours after the first exposure and 25 hours after the in vivo labelling of DNA. A parallelism has been found between the results of this experiment and the recovery effect of the Elkind-Sutton type. It may be concluded that the dying process responsible for the loss of the dividing cells from thymus and spleen after irradiation probably involves reproductive death.

Isotope dilution mass spectrometry for procaine determination in biological samples.

A simple, rapid and sensitive method for procaine determination is described. Isotope dilution mass spectrometry with (15N)procaine as internal standard was used. The analysis was performed at 4000 resolution by selected ion monitoring with temperature programming. The sample was measured in underivatized form in the direct inlet system. The method shows good analytical parameters: linearity between 0 and 40 micrograms ml-1, good precision and accuracy. The method was applied to the in vitro pharmacokinetic study of the metabolism of procaine in liver homogenates of Wistar rats. The method is rapid, permitting about six samples to be run per hour. Sensitivity of the method permits analysis at a signal-to-noise ratio of 5:1.