Transgendered in Alaska: Navigating the Changing Legal Landscape for Change in Gender Petitions

Background: Detecting intracellular bacterial symbionts can be challenging when they persist at very low densities. Wolbachia, a widespread bacterial endosymbiont of invertebrates, is particularly challenging. Although it persists at high titers in many species, in others its densities are far below the detection limit of classic end-point Polymerase Chain Reaction (PCR). These low-titer infections can be reliably detected by combining PCR with DNA hybridization, but less elaborate strategies based on end-point PCR alone have proven less sensitive or less general. Results: We introduce a multicopy PCR target that allows fast and reliable detection of A-supergroup Wolbachia -even at low infection titers -with standard end-point PCR. The target is a multicopy motif (designated ARM: A-supergroup repeat motif) discovered in the genome of wMel (the Wolbachia in Drosophila melanogaster). ARM is found in at least seven other Wolbachia A-supergroup strains infecting various Drosophila, the wasp Muscidifurax and the tsetse fly Glossina. We demonstrate that end-point PCR targeting ARM can reliably detect both high-and low-titer Wolbachia infections in Drosophila, Glossina and interspecific hybrids. Conclusions: Simple end-point PCR of ARM facilitates detection of low-titer Wolbachia A-supergroup infections. Detecting these infections previously required more elaborate procedures. Our ARM target seems to be a general feature of Wolbachia A-supergroup genomes, unlike other multicopy markers such as insertion sequences (IS)

Identification and characterization of Vitis vinifera subsp sylvestris populations in north-western Italy

The Italian peninsula, for its favorable environmental and geo-morphological conditions, can be considered an ultimate area for survival and development of Vitis vinifera subsp. sylvestris (Gmelin) Hegi, even though severely affected by human impact. Large surveys started in early 1990 throughout the country. At the time few regions, like Piedmont located in the north-west of the country, were considered lacking of wild vinifera. More recent prospection started several years ago, leading to the discovering of five vinifera sylvestris populations plus other sites with few individuals. The sites of discovery were described for their ecological features and the identified plants were referenced and characterized by morphology (18 descriptors from the OIV list) and genetics (14 n-SSR loci). The esteemed consistency of each population ranged from 20 to 150 individuals. Morphological and biological traits (dioecious plants, females producing very small roundish black berries), as well as genetic profiles, indicated the observed plants are true vinifera sylvestris. As to the ecological requirements, plants were confirmed to be highly dependent on water availability into the soil. The neighbor-joining (NJ) dendrogram resulting from SSR allelic pattern of the individuals belonging to the five populations and to one location with isolated plants, indicated population's genetic similarity broadly reflects site's geographic distance. Considering the numerous reports in the past, spreading and consistency of wild grape germplasm from the region of Piedmont severely decreased over a period of 100-150 years. The relative short distance from wild population's sites and vineyards must also be regarded as a worrying condition because of contamination risks. All means to avoid the loss of this native Vitis germplasm must be undertaken by protection policy and proper land management

Spatio-temporal distribution of Spiroplasma infections in the tsetse fly (Glossina fuscipes fuscipes) in northern Uganda

Copyright: © 2019 Schneider et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Tsetse flies (Glossina spp.) are vectors of parasitic trypanosomes, which cause human (HAT) and animal African trypanosomiasis (AAT) in sub-Saharan Africa. In Uganda, Glossina fuscipes fuscipes (Gff) is the main vector of HAT, where it transmits Gambiense disease in the northwest and Rhodesiense disease in central, southeast and western regions. Endosymbionts can influence transmission efficiency of parasites through their insect vectors via conferring a protective effect against the parasite. It is known that the bacterium Spiroplasma is capable of protecting its Drosophila host from infection with a parasitic nematode. This endosymbiont can also impact its host\u27s population structure via altering host reproductive traits. Here, we used field collections across 26 different Gff sampling sites in northern and western Uganda to investigate the association of Spiroplasma with geographic origin, seasonal conditions, Gff genetic background and sex, and trypanosome infection status. We also investigated the influence of Spiroplasma on Gff vector competence to trypanosome infections under laboratory conditions. Generalized linear models (GLM) showed that Spiroplasma probability was correlated with the geographic origin of Gff host and with the season of collection, with higher prevalence found in flies within the Albert Nile (0.42 vs 0.16) and Achwa River (0.36 vs 0.08) watersheds and with higher prevalence detected in flies collected in the intermediate than wet season. In contrast, there was no significant correlation of Spiroplasma prevalence with Gff host genetic background or sex once geographic origin was accounted for in generalized linear models. Additionally, we found a potential negative correlation of Spiroplasma with trypanosome infection, with only 2% of Spiroplasma infected flies harboring trypanosome co-infections. We also found that in a laboratory line of Gff, parasitic trypanosomes are less likely to colonize the midgut in individuals that harbor Spiroplasma infection. These results indicate that Spiroplasma infections in tsetse may be maintained by not only maternal but also via horizontal transmission routes, and Spiroplasma infections may also have important effects on trypanosome transmission efficiency of the host tsetse. Potential functional effects of Spiroplasma infection in Gff could have impacts on vector control approaches to reduce trypanosome infections

Single fraction radiosurgery using Rapid Arc for treatment of intracranial targets

<p>Abstract</p> <p>Background</p> <p>Stereotactic-Radio-Surgery (SRS) using Conformal-Arc-Therapy (CAT) is a well established irradiation technique for treatment of intracranial targets. Although small safety margins are required because of very high accuracy of patient positioning and exact online localisation, there are still disadvantages like long treatment time, high number of monitor units (MU) and covering of noncircular targets. This planning study analysed whether Rapid Arc (RA) with stereotactic localisation for single-fraction SRS can solve these problems.</p> <p>Methods</p> <p>Ten consecutive patients were treated with Linac-based SRS. Eight patients had one or more brain metastases. The other patients presented a symptomatic vestibularis schwannoma and an atypic meningeoma. For all patients, two plans (CAT/RA) were calculated and analysed.</p> <p>Results</p> <p>Conformity was higher for RA with additional larger low-dose areas. Furthermore, RA reduced the number of MU and the treatment time for all patients. Dose to organs at risk were equal or slightly higher using RA in comparison to CAT.</p> <p>Conclusions</p> <p>RA provides a new alternative for single-fraction SRS irradiation combining advantages of short treatment time with lower number of MU and better conformity in addition to accuracy of stereotactic localisation in selected cases with uncomplicated clinical realization.</p

Dynamics of the extremely elongated cloud on Mars Arsia Mons volcano

Starting in September 2018, a daily repeating extremely elongated cloud was observed extending up to 1800km from the Mars Arsia Mons volcano. We study this Arsia Mons Elongated Cloud (AMEC) using images from VMC, HRSC, and OMEGA on board Mars Express, IUVS on MAVEN, MCC on Mars Orbiter Mission (MOM), MARCI on MRO, and Visible Camera on Viking 2 orbiter. We study the daily cycle of this cloud, showing how the morphology and other parameters of the cloud evolved rapidly with local time. The cloud expands every morning from the western slope of the volcano, at a westward velocity of around 160m/s, and an altitude of around 45km over martian areoid. The expansion starts with sunrise, and resumes around 2.5 hours later, when cloud formationresumes and the elongated tail detaches from the volcano and keeps moving westward until it evaporates before afternoon, when most sun-synchronous missions observe. This daily cycle repeated regularly for at least 80 sols in 2018 (Martian Year 34). We find in images from past years that this AMEC is an annually repeating phenomenon that takes place around the Solar Longitude range 220º-320º. We study the AMEC in Martian Year 34 in terms of Local Time and Solar Longitude, and then compare with observations from previous years, in search for interannual variations, taking into account the possible influence of the recent Global Dust Storm

Dynamics of the extremely elongated cloud on Mars Arsia Mons volcano

Starting in September 2018, a daily repeating extremely elongated cloud was observed extending from the Mars Arsia Mons volcano. We study this Arsia Mons Elongated Cloud (AMEC) using images from VMC, HRSC, and OMEGA on board Mars Express, IUVS on MAVEN, and MARCI on MRO. We study the daily cycle of this cloud, showing how the morphology and other parameters of the cloud evolved with local time. The cloud expands every morning from the western slope of the volcano, at a westward velocity of around 150m/s, and an altitude of around 30-40km over the local surface. Starting around 2.5 hours after sunrise (8.2 Local True Solar Time, LTST), the formation of the cloud resumes, and the existing cloud keeps moving westward, so it detaches from the volcano, until it evaporates in the following hours. At this time, the cloud has expanded to a length of around 1500km. Short time later, a new local cloud appears on the western slope of the volcano, starting around 9.5 LTST, and grows during the morning. This daily cycle repeated regularly for at least 90 sols in 2018, around Southern Solstice (Ls 240-300) in Martian Year (MY) 34. According with these and previous MEx/VMC observations, this elongated cloud is a seasonal phenomenon occurring around Southern Solstice every Martian Year. We study the interannual variability of this cloud, the influence of the Global Dust Storms in 2018 on the cloud’s properties (Sánchez-Lavega et al., Geophys. Res. Lett. 46, 2019), and its validity as a proxy for the global state of the Martian atmosphere (Sánchez-Lavega et al., J. Geophys. Res., 123, 3020, 2018). We discuss the physical mechanisms behind the formation of this peculiar cloud in Mars

mTORC1 is essential for early steps during Schwann cell differentiation of amniotic fluid stem cells and regulates lipogenic gene expression.

Schwann cell development is hallmarked by the induction of a lipogenic profile. Here we used amniotic fluid stem (AFS) cells and focused on the mechanisms occurring during early steps of differentiation along the Schwann cell lineage. Therefore, we initiated Schwann cell differentiation in AFS cells and monitored as well as modulated the activity of the mechanistic target of rapamycin (mTOR) pathway, the major regulator of anabolic processes. Our results show that mTOR complex 1 (mTORC1) activity is essential for glial marker expression and expression of Sterol Regulatory Element-Binding Protein (SREBP) target genes. Moreover, SREBP target gene activation by statin treatment promoted lipogenic gene expression, induced mTORC1 activation and stimulated Schwann cell differentiation. To investigate mTORC1 downstream signaling we expressed a mutant S6K1, which subsequently induced the expression of the Schwann cell marker S100b, but did not affect lipogenic gene expression. This suggests that S6K1 dependent and independent pathways downstream of mTORC1 drive AFS cells to early Schwann cell differentiation and lipogenic gene expression. In conclusion our results propose that future strategies for peripheral nervous system regeneration will depend on ways to efficiently induce the mTORC1 pathway

Identification and in vitro Analysis of the GatD/MurT Enzyme-Complex Catalyzing Lipid II Amidation in Staphylococcus aureus

The peptidoglycan of Staphylococcus aureus is characterized by a high degree of crosslinking and almost completely lacks free carboxyl groups, due to amidation of the D-glutamic acid in the stem peptide. Amidation of peptidoglycan has been proposed to play a decisive role in polymerization of cell wall building blocks, correlating with the crosslinking of neighboring peptidoglycan stem peptides. Mutants with a reduced degree of amidation are less viable and show increased susceptibility to methicillin. We identified the enzymes catalyzing the formation of D-glutamine in position 2 of the stem peptide. We provide biochemical evidence that the reaction is catalyzed by a glutamine amidotransferase-like protein and a Mur ligase homologue, encoded by SA1707 and SA1708, respectively. Both proteins, for which we propose the designation GatD and MurT, are required for amidation and appear to form a physically stable bi-enzyme complex. To investigate the reaction in vitro we purified recombinant GatD and MurT His-tag fusion proteins and their potential substrates, i.e. UDP-MurNAc-pentapeptide, as well as the membrane-bound cell wall precursors lipid I, lipid II and lipid II-Gly5. In vitro amidation occurred with all bactoprenol-bound intermediates, suggesting that in vivo lipid II and/or lipid II-Gly5 may be substrates for GatD/MurT. Inactivation of the GatD active site abolished lipid II amidation. Both, murT and gatD are organized in an operon and are essential genes of S. aureus. BLAST analysis revealed the presence of homologous transcriptional units in a number of gram-positive pathogens, e.g. Mycobacterium tuberculosis, Streptococcus pneumonia and Clostridium perfringens, all known to have a D-iso-glutamine containing PG. A less negatively charged PG reduces susceptibility towards defensins and may play a general role in innate immune signaling

Canine distemper virus induces apoptosis in cervical tumor derived cell lines

Apoptosis can be induced or inhibited by viral proteins, it can form part of the host defense against virus infection, or it can be a mechanism for viral spread to neighboring cells. Canine distemper virus (CDV) induces apoptotic cells in lymphoid tissues and in the cerebellum of dogs naturally infected. CDV also produces a cytopathologic effect, leading to apoptosis in Vero cells in tissue culture. We tested canine distemper virus, a member of the Paramyxoviridae family, for the ability to trigger apoptosis in HeLa cells, derived from cervical cancer cells resistant to apoptosis. To study the effect of CDV infection in HeLa cells, we examined apoptotic markers 24 h post infection (pi), by flow cytometry assay for DNA fragmentation, real-time PCR assay for caspase-3 and caspase-8 mRNA expression, and by caspase-3 and -8 immunocytochemistry. Flow cytometry showed that DNA fragmentation was induced in HeLa cells infected by CDV, and immunocytochemistry revealed a significant increase in the levels of the cleaved active form of caspase-3 protein, but did not show any difference in expression of caspase-8, indicating an intrinsic apoptotic pathway. Confirming this observation, expression of caspase-3 mRNA was higher in CDV infected HeLa cells than control cells; however, there was no statistically significant change in caspase-8 mRNA expression profile. Our data suggest that canine distemper virus induced apoptosis in HeLa cells, triggering apoptosis by the intrinsic pathway, with no participation of the initiator caspase -8 from the extrinsic pathway. In conclusion, the cellular stress caused by CDV infection of HeLa cells, leading to apoptosis, can be used as a tool in future research for cervical cancer treatment and control