Responding to eruptive transitions during the 2020-2021 eruption of La Soufrière volcano, St. Vincent.

A critical challenge during volcanic emergencies is responding to rapid changes in eruptive behaviour. Actionable advice, essential in times of rising uncertainty, demands the rapid synthesis and communication of multiple datasets with prognoses. The 2020-2021 eruption of La Soufrière volcano exemplifies these challenges: a series of explosions from 9-22 April 2021 was preceded by three months of effusive activity, which commenced with a remarkably low level of detected unrest. Here we show how the development of an evolving conceptual model, and the expression of uncertainties via both elicitation and scenarios associated with this model, were key to anticipating this transition. This not only required input from multiple monitoring datasets but contextualisation via state-of-the-art hazard assessments, and evidence-based knowledge of critical decision-making timescales and community needs. In addition, we share strategies employed as a consequence of constraints on recognising and responding to eruptive transitions in a resource-constrained setting, which may guide similarly challenged volcano observatories worldwide.

Anomalous hall heat current and nernst effect in the CuCr2Se4-xBrx ferromagnet.

In a ferromagnet, an anomalous Hall heat current, given by the off-diagonal Peltier term alpha(xy), accompanies the anomalous Hall current. By combining Nernst, thermopower, and Hall experiments, we have measured how alpha(xy) varies with hole density and lifetime tau in CuCr2Se4-xBrx. At low temperatures T, we find that alpha(xy) is independent of tau, consistent with anomalous-velocity theories. Its magnitude is fixed by a microscopic geometric area A approximately 34 A(2). Our results are incompatible with some models of the Nernst effect in ferromagnets.

Dissipationless anomalous Hall current in the ferromagnetic spinel CuCr2Se4-xBrx.

In a ferromagnet, an applied electric field E invariably produces an anomalous Hall current JH that flows perpendicular to the plane defined by E and M (the magnetization). For decades, the question of whether JH is dissipationless (independent of the scattering rate) has been debated without experimental resolution. In the ferromagnetic spinel CuCr2Se4-xBrx, the resistivity rho (at low temperature) may be increased by several decades by varying x (Br) without degrading M. We show that JH/E (normalized per carrier, at 5 kelvin) remains unchanged throughout. In addition to confirming the dissipationless nature of JH, our finding has implications for the generation and study of spin-Hall currents in bulk samples.

Yersinia virulence: more than a plasmid.

The genus Yersinia is composed of 11 species, three of which are pathogenic in humans. The three pathogens, Y. pestis, Y. enterocolitica, and Y. pseudotuberculosis, cause a broad spectrum of disease ranging from pneumonic plague to acute gastroenteritis. Each of the three requires a large, well-defined plasmid for full virulence, as well as many chromosomally encoded virulence factors (CEVF). This review will describe these CEVF and their roles in virulence. In addition, a possible model for key events in Y. enterocolitica pathogenesis is described based on information revealed by analysis of several of the CEVF.

Two-step chromatographic purification of recombinant Plasmodium falciparum circumsporozoite protein from Escherichia coli.

The Plasmodium falciparum circumsporozoite (PfCS) protein (aa 19-405) has been cloned and expressed in E. coli. The protein was purified in a two-step process that was rapid and reproducible. E. coli cells were grown to a high density before induction for 1 h. Cells were disrupted by high pressure microfluidization and the total bacterial protein solubilized in 6 M Gu-HCl. The protein was refolded while bound to Ni-NTA agarose by exchange of 6 M Gu-HCl for 8 M urea and then slow removal of the urea. The eluted protein was further purified on Q Sepharose Fast Flow using conditions developed to remove E. coli proteins and reduce endotoxin (to 10 EU/50 microg). Yield was 20 mg of PfCS protein from 10 g of wet cell paste. The final protein product bound to HepG2 liver cells in culture and inhibited the invasion of those cells by sporozoites in an ISI assay greater than 80% over control cultures when used at 10 microg/ml.

Identification of regions of Ail required for the invasion and serum resistance phenotypes.

Yersinia enterocolitica is an enteric pathogen that has served as a model system for the study of microbial pathogenesis. Numerous virulence gene have been identified both on the virulence plasmid and on the chromosome. One of the chromosomal genes that is highly correlated with virulence is ail, a gene identified along with inv in a screen for Y. enterocolitica genes that could confer an invasive phenotype to Escherichia coli. Ail also promotes serum resistance in both E. coli and Y. enterocolitica. Several virulence factors homologous to Ail have been identified in other pathogens, yet very little is known about what constitutes the functional domain(s) of these proteins. Proteins in this family are predicted to consist of eight transmembrane beta-sheets and four cell surface-exposed loops. We constructed and characterized a number of insertion, deletion and point mutations in the regions of ail predicted to encode the cell surface loops. The results from the analysis of these mutants indicate that cell surface loops one and four do not directly promote invasion or serum resistance, whereas mutations in loop three appear to modulate both phenotypes. Analysis of mutations in loop 2 suggests that this surface-exposed loop contains sequences required for serum resistance and invasion. In addition, a peptide derived from the sequence of loop 2 was able specifically to inhibit Ail-mediated invasion in a dose-dependent manner. These results suggest that Ail directly promotes invasion and that loop 2 contains an active site, perhaps a receptor-binding domain. Analyses of the mutations also suggest that the serum resistance and invasion phenotypes may be separable, because there are numerous mutations that affect one phenotype but not the other.

Identification of a locus involved in systemic dissemination of Yersinia enterocolitica.

A putative LysR-type transcriptional activator, Hre20, was identified previously in an in vivo expression technology screen designed to identify factors which are expressed early during infection by Yersinia enterocolitica (G. M. Young and V. L. Miller, Mol. Microbiol. 25:319-328, 1997). An insertion in hre20, now designated rscR, resulted in increased splenic dissemination of bacteria during infection in a BALB/c mouse model. A nonpolar mutation was generated in rscR, and examination of this strain in the BALB/c mouse model demonstrated that the mutation in rscR was responsible for the increased dissemination to the spleen that was seen in the original experiments. RscR is homologous to the LysR family of transcriptional regulators; thus, a screen was undertaken to identify genes regulated by RscR. A strain containing an insertion in the chromosomal rscR gene and carrying rscR on a plasmid under the control of the inducible araBAD promoter was mutagenized with an mTn5Km-2 transposon containing a promoterless lacZY. Eighteen insertions were identified which appeared to respond to levels of RscR, and these were classified into four allelic groups based on Southern blot hybridization analysis. Representative members were sequenced from three allelic groups. Sequencing revealed insertions in an ORF with no known homologues, a homologue of OmpF of Serratia marcescens, and a locus (designated rscBAC) with similarity to the hmwABC locus of Haemophilus influenzae. The hmwABC locus promotes adherence of H. influenzae to host cells (S. J. Barenkamp and J. W. St. Geme III, Infect. Immun. 62:3320-3328, 1994; J. W. St. Geme III, S. Falkow, and S. J. Barenkamp, Proc. Natl. Acad. Sci. USA 90:2875-2879, 1993). A strain containing a deletion mutant of rscA, the hmwA homologue, exhibits increased splenic dissemination of bacteria during infection in a BALB/c mouse model, similar to the rscR mutant. This suggests that the phenotype of an rscR mutant is due to the loss of RscA.

A role for IL-1 alpha in inducing pathologic inflammation during bacterial infection.

Infection with pathogenic microbes often results in a significant inflammatory response. A cascade of proinflammatory cytokines including tumor necrosis factor alpha (TNF-alpha) and IL-1 initiates this response. Although there is a clear role for IL-1 during infection, little is known to distinguish the role of IL-1 alpha from that of IL-1 beta during this process. With the use of Yersinia enterocolitica as a model enteric pathogen, we have identified a specific role for IL-1 alpha in inducing pathologic inflammation during bacterial infection. Depletion of IL-1 alpha in mice infected with wild-type Y. enterocolitica results in significantly decreased intestinal inflammation. Furthermore, a bacterial mutant that does not induce IL-1 alpha expression but induces normal levels of IL-1 beta, TNF-alpha, and IFN-gamma, causes greatly reduced intestinal inflammation and is attenuated by LD(50) analysis in the C57BL/6 mouse model. These results demonstrate a distinct and unrecognized role for IL-1 alpha in inducing intestinal inflammation that cannot be compensated for by the endogenous levels of IL-1 beta, TNF-alpha, or IFN-gamma that are produced in response to Y. enterocolitica. Additionally, these results suggest that IL-1 alpha-induced inflammation is a major contributor to the pathology of yersiniosis.

A comparison of heat-induced hyperactivation in patients' sperm after colloid or pentoxifylline wash methods.

OBJECTIVE: Our purpose was to compare kinematic parameters of human sperm after processing through two different wash methods and 40 degrees C heat treatment. STUDY DESIGN: Sperm specimens (N = 169 cases) were washed by either colloid or pentoxifylline wash methods, and the motility parameters were measured at either 37 degrees C or 40 degrees C at baseline (0 hours) and after 4 hours. Five randomly selected washed specimens with matching 37 degrees C (control) or 40 degrees C heat treatments were assessed for changes in a sentinel gene. RESULTS: The percentage of sperm hyperactive motility was >5 times higher after the 40 degrees C heat treatment, in comparison with the 37 degrees C treatment, for both the colloid- and the pentoxifylline-washed sperm. The percentages of total motility and progression were equally enhanced in heated sperm for the two wash methods. No changes were detected in the sentinel gene with the heat treatment. CONCLUSION: Sperm cells mildly heated at 40 degrees C responded with greater motility, progression, and hyperactivation. The data suggest that mild heat is a stimulus for sperm function because greater sperm hyperactivation is associated with increased sperm fertilizing capacity. The absence of change in the sentinel gene in heated sperm suggests that a temperature of 40 degrees C is too low to initiate alterations in the highly condensed sperm chromatin. More studies are needed before mild heating of ejaculated sperm becomes acceptable for use in assisted reproductive technologies.

HreP, an in vivo-expressed protease of Yersinia enterocolitica, is a new member of the family of subtilisin/kexin-like proteases.

The role of proteases in pathogenesis is well established for several microorganisms but has not been described for Yersinia enterocolitica. Previously, we identified a gene, hreP, which showed significant similarity to proteases in a screen for chromosomal genes of Y. enterocolitica that were exclusively expressed during an infection of mice. We cloned this gene by chromosome capture and subsequently determined its nucleotide sequence. Like inv, the gene encoding the invasin protein of Y. enterocolitica, hreP is located in a cluster of flagellum biosynthesis and chemotaxis genes. The genomic organization of this chromosomal region is different in Escherichia coli, Salmonella, and Yersinia pestis than in Y. enterocolitica. Analysis of the distribution of hreP between different Yersinia isolates and the relatively low G+C content of the gene suggests acquisition by horizontal gene transfer. Sequence analysis also revealed that HreP belongs to a family of eukaryotic subtilisin/kexin-like proteases. Together with the calcium-dependent protease PrcA of Anabaena variabilis, HreP forms a new subfamily of bacterial subtilisin/kexin-like proteases which might have originated from a common eukaryotic ancestor. Like other proteases of this family, HreP is expressed with an N-terminal prosequence. Expression of an HreP-His(6) tag fusion protein in E. coli revealed that HreP undergoes autocatalytic processing at a consensus cleavage site of subtilisin/kexin-like proteases, thereby releasing the proprotein.

Type III secretion chaperone-dependent regulation: activation of virulence genes by SicA and InvF in Salmonella typhimurium.

Invasion of the intestinal epithelium by Salmonella sp. requires a type III secretion system (TTSS) common in many bacterial pathogens. TTSS translocate effector proteins from bacteria into eukaryotic cells. These effectors manipulate cellular functions in order to benefit the pathogen. In the human and animal pathogen Salmonella typhimurium, the expression of genes encoding the secreted effector molecules Sip/Ssp ABCD, SigD, SptP and SopE requires both the AraC/XylS-like regulator InvF and the secretion chaperone SICA: In this work, an InvF binding site was identified in the promoter regions of three operons. SicA does not appear to affect InvF stability nor to bind DNA directly. However, SicA could be co-purified with InvF, suggesting that InvF and SicA interact with each other to activate transcription from the effector gene promoters. This is the first demonstration of a contact between a protein cofactor and an AraC/XylS family transcriptional regulator and, moreover, is the first direct evidence of an interaction between a transcriptional regulator and a TTSS chaperone. The regulation of effector genes described here for InvF and SicA may represent a new paradigm for regulation of virulence in a wide variety of pathogens.

SigE is a chaperone for the Salmonella enterica serovar Typhimurium invasion protein SigD.

SigD is translocated into eucaryotic cells by a type III secretion system. In this work, evidence that the putative chaperone SigE directly interacts with SigD is presented. A bacterial two-hybrid system demonstrated that SigE can interact with itself and SigD. In addition, SigD was specifically copurified with SigE-His(6) on a nickel column.

The psp locus of Yersinia enterocolitica is required for virulence and for growth in vitro when the Ysc type III secretion system is produced.

The phage shock protein locus (pspFpspABCDE) of Escherichia coli has proved to be something of an enigma since its discovery. The physiological functions of the psp locus, including those of the predicted effector protein PspA, are unknown. In a previous genetic screen, we determined that a Yersinia enterocolitica pspC mutant was severely attenuated for virulence. In this study, the psp locus of Y. enterocolitica was characterized further. The pspC gene of Y. enterocolitica was found to be important for normal growth when the Ysc type III secretion system was expressed in the laboratory. This growth defect was specifically caused by production of the secretin protein, YscC. Expression of the psp genes was induced when the type III secretion system was functional or when only the yscC gene was expressed. This induction of psp gene expression required a functional pspC gene. Most significantly, evidence suggests that the expression of at least one gene that is not part of the psp locus is regulated by Psp proteins. This unidentified gene (or genes) may also be important for growth when the type III secretion system is expressed. These conclusions are supported by the effects of various psp mutations on virulence. This is the first indication that Psp proteins might be involved in the regulation of genes besides the psp locus itself.

Contraceptive decision making among Medicaid-eligible women.

The purpose of this study was to examine the influence of sociodemographic factors, attitudes, knowledge, and experiences regarding the use of various contraceptive methods, future plans prenatally, and actual use postpartum among a population of low income pregnant women. Women were interviewed prenatally and during the postpartum period in a large, urban academic health center serving primarily an indigent population. The primary analytic method employed was logistic regression. The key finding in this study is that women are not consistently using the method of contraception postpartum that they planned during the prenatal period. Only 54.7% of the women planning to use oral contraceptive pills were using them postpartum, and only 31.3% of the women planning to use condoms were actually using them postpartum. Expanding contraceptive education and counseling throughout the perinatal period may assist women's decision making.

Identification and characterization of Yersinia enterocolitica genes induced during systemic infection.

Yersinia enterocolitica is one of three pathogenic Yersinia species that share a tropism for lymphoid tissues. However, infection of an immunocompromised host is likely to result in a systemic infection, which is often fatal. Little is known about the bacterial proteins needed to establish such an infection. The genes that encode these virulence factors are likely to be active only during systemic infection. A library of random cat fusions was used to inoculate BALB/c mice. Fusions expressed during a systemic infection were enriched by the administration of chloramphenicol-succinate. Y. enterocolitica isolates recovered from the mice were tested for chloramphenicol resistance in vitro. Fusions that were inactive in vitro were analyzed further and found to represent 31 allelic groups. Each was given a sif (for systemic infection factor) designation. Based on homology to known proteins, the sif genes are likely to encode proteins important for general physiology, transcription regulation, and other functions. During systemic infections, 13 of the sif-cat fusions were able to outcompete the wild type in the presence of chloramphenicol-succinate, confirming that the fusions were active. The in vitro expression of several sif genes was determined, showing modest changes in response to various growth conditions. A mutation in sif15, which encodes a putative outer membrane protein, caused attenuation during systemic infection but not during colonization of the Peyer's patches. Comparisons between the Y. enterocolitica sif genes and the previously identified hre genes imply that very different groups of genes are active during a systemic infection and during colonization of the Peyer's patches.

Yersinia enterocolitica ClpB affects levels of invasin and motility.

Expression of the Yersinia enterocolitica inv gene is dependent on growth phase and temperature. inv is maximally expressed at 23 degrees C in late-exponential- to early-stationary-phase cultures. We previously reported the isolation of a Y. enterocolitica mutant (JB1A8v) that shows a decrease in invasin levels yet is hypermotile when grown at 23 degrees C. JB1A8v has a transposon insertion within uvrC. Described here is the isolation and characterization of a clone that suppresses these mutant phenotypes of the uvrC mutant JB1A8v. This suppressing clone encodes ClpB (a Clp ATPase homologue). The Y. enterocolitica ClpB homologue is 30 to 40% identical to the ClpB proteins from various bacteria but is 80% identical to one of the two ClpB homologues of Yersinia pestis. A clpB::TnMax2 insertion mutant (JB69Qv) was constructed and determined to be deficient in invasin production and nonmotile when grown at 23 degrees C. Analysis of inv and fleB (flagellin gene) transcript levels in JB69Qv suggested that ClpB has both transcriptional and posttranscriptional effects. In contrast, a clpB null mutant, BY1v, had no effect on invasin levels or motility. A model accounting for these observations is presented.

Motility is required to initiate host cell invasion by Yersinia enterocolitica.

Invasin-mediated invasion of host cells by the pathogen Yersinia enterocolitica was shown to be affected by flagellar-dependent motility. Motility appears to be required to ensure the bacterium migrates to and contacts the host cell. Nonmotile strains of Y. enterocolitica were less invasive than motile strains, but the reduction in invasion could be overcome by artificially bringing the bacteria into host cell contact by centrifugation. Mutations in known regulatory genes of the flagellar regulon, flhDC and fliA, resulted in less inv expression but did not have a significant effect on invasin levels. However, invasin levels were reduced for strains that harbored flhDC on a multicopy plasmid, apparently as a result of increased proteolysis of invasin.

Multifetal pregnancy reduction: perinatal and fiscal outcomes.

OBJECTIVE: This study was undertaken to compare the birth outcomes of a multifetal pregnancy reduction population with those of other patients delivered at Hutzel Hospital, Detroit, and to determine the fiscal impact of the multifetal pregnancy reduction program. STUDY DESIGN: In a retrospective review patients who were delivered after multifetal pregnancy reduction were compared with a general obstetric population who were delivered at Hutzel Hospital from January 1, 1986, through June 30, 1998. Outcome data were determined through a comprehensive perinatal database. The chi(2) analysis was used to examine the relationship between gestational age and delivery group. Financial data were estimated from published reports of neonatal intensive care unit admissions, cost estimates for neonatal intensive care unit care, and charges for multifetal pregnancy reduction. RESULTS: Pregnancies reduced to triplets, twins, and singletons had outcomes at least comparable to unreduced pregnancies starting at these numbers and substantially better than unreduced pregnancies with the same starting number. Financial estimates of hospitalization costs averted in the multifetal pregnancy reduction population exceeded $28 million. CONCLUSION: Use of multifetal pregnancy reduction improved obstetric outcomes for pregnancies with multiple gestations and also was associated with significant fiscal savings.

Fiscal impact of a potential legislative ban on second trimester elective terminations for prenatally diagnosed abnormalities.

This study was designed to determine the fiscal impact of a theoretical legislative ban on elective terminations for prenatally diagnosed abnormalities at Hutzel Hospital/Wayne State University. A fiscal comparison was completed for patients who had second trimester elective terminations for prenatally diagnosed abnormalities versus not allowing the procedure. An eight-year database of genetics cases and hospital and physician cost estimates for performing elective terminations for prenatally diagnosed abnormalities, and published reports of the average lifetime costs per selected birth defects, were used to calculate the net cost. The estimated lifetime cost for an average cohort year of a legislative ban on elective terminations for prenatally diagnosed abnormalities was found to be at least $8.5 million for patients treated at Hutzel Hospital. Extrapolated, a similar ban on second trimester elective terminations would have a net cost of $74 million in Michigan and $2 billion annually in the United States.

The Yersinia enterocolitica phospholipase gene yplA is part of the flagellar regulon.

Yersinia enterocolitica yplA encodes a phospholipase required for virulence. Virulence genes are often regulated in response to environmental signals; therefore, yplA expression was examined using a yplA::lacZY transcriptional fusion. Maximal yplA expression occurred between pH 6.5 and pH 7.5 and was induced in the mid-logarithmic growth phase. Potential Fnr, cyclic AMP (cAMP)-cAMP receptor protein (Crp), and sigma(F) regulatory sites were identified in the nucleotide sequence. Reduction of yplA expression by aeration, addition of glucose and sucrose, and application of high temperature and salt is consistent with Fnr-, cAMP-Crp-, and sigma(F)-mediated regulation, respectively. Expression of yplA was reduced in flhDC and fliA null strains, indicating that yplA is part of the flagellar regulon.