A plague epizootic in the white-tailed prairie dogs (Cynomys leucurus) of Meeteetse, Wyoming

Surveillance for sylvatic plague (Yersinia pestis) was conducted near Meeteetse, Wyoming (USA) from 24 May to 14 June 1985. Ten species of fleas were collected from white-tailed prairie dogs (Cynomys leucurus), and from their burrows and associated rodents. Five of these flea species and two adult prairie dogs were positive for plague. The progression of this plague epizootic appeared to be slower and the intensity was less than in previous epizootics in other prairie dog colonies. The plague epizootic occurred within the only known colony of black-footed ferrets (Mustela nigripes) and was a potential threat to the food source of this endangered species.     

Superfluous killing in spiders: a consequence of adaptation to food-limited environments?

The hypothesis that superfluous killing, partial consumption,and abandonment of prey is a consequence of adaptation to food-limited environments was tested in two feeding trials on a desert spider, Agelenopsis aperta. First, we made comparisons among populations inhabiting sites of high prey (HP) or low prey (LP) availabilitythat differed in their degree of genetic isolation. Typically,A. aperta entirely consumed one or two of the prey items itcaptured in a feeding bout. Additional prey were partiallyconsumed or abandoned without eating. Spiders from the geneticallyisolated HP population, however, captured fewer prey and showeda higher incidence of full feeding on prey than did individualsfrom the other populations. Only one spider from this populationcaptured a prey item that it failed to feed on, whereas spidersfrom LP populations failed to feed on high numbers of capturedprey. The greatest variability in feeding behavior was exhibitedin the HP population that experienced gene flow. The secondtest was based on the finding that aggressiveness is largelya sex-linked trait in A. aperta: the aggressiveness of thefemale parent only is inherited by male offspring, whereasboth parents contribute to this trait in female offspring.All female F₁ hybrids between LP and HP parental types exhibitedhigh levels of superfluous killing, as did male F₁ hybridsderived from LP females. F₁ hybrid males derived from HP femalesexhibited extremely low levels of superfluous killing. Superfluouskilling thus has its basis in the genetic control of levelsof aggression.     

Disulfide bond of Mycoplasma pneumoniae community‐acquired respiratory distress syndrome toxin is essential to maintain the ADP‐ribosylating and vacuolating activities

Mycoplasma pneumoniae is the leading cause of bacterial community‐acquired pneumonia among hospitalised children in United States and worldwide. Community‐acquired respiratory distress syndrome (CARDS) toxin is a key virulence determinant of M. pneumoniae. The N‐terminus of CARDS toxin exhibits ADP‐ribosyltransferase (ADPRT) activity, and the C‐terminus possesses binding and vacuolating activities. Thiol‐trapping experiments of wild‐type (WT) and cysteine‐to‐serine‐mutated CARDS toxins with alkylating agents identified disulfide bond formation at the amino terminal cysteine residues C230 and C247. Compared with WT and other mutant toxins, C247S was unstable and unusable for comparative studies. Although there were no significant variations in binding, entry, and retrograde trafficking patterns of WT and mutated toxins, C230S did not elicit vacuole formation in intoxicated cells. In addition, the ADPRT domain of C230S was more sensitive to all tested proteases when compared with WT toxin. Despite its in vitro ADPRT activity, the reduction of C230S CARDS toxin‐mediated ADPRT activity‐associated IL‐1β production in U937 cells and the recovery of vacuolating activity in the protease‐released carboxy region of C230S indicated that the disulfide bond was essential not only to maintain the conformational stability of CARDS toxin but also to properly execute its cytopathic effects.     

Preferred orientations of His64 in human carbonic anhydrase II

Histidine at position 64 (His64) in human carbonic anhydrase II (HCA II) is believed to be the proton acceptor in the hydration direction and the proton donor in the dehydration direction for the rate-limiting proton transfer (PT) event. Although the biochemical effect of histidine at position 64 has been thoroughly investigated, the role of its orientation in the PT event is a topic of considerable debate. X-ray data of HCA II suggests that His64 can adopt either an "in" or "out" orientation. The "in" orientation is believed to be favored for the hydration direction PT event because the Ndelta of His64 is closer to the catalytic zinc. This orientation allows for smaller water bridges, which are postulated to be more conducive to PT. In the present work, classical molecular dynamics simulations have been conducted to elucidate the role that the His64 orientation may play in its ability to act as a proton donor/acceptor in HCA II. The free energy profile for the orientation of His64 suggests that the histidine will adopt an "in" orientation in the hydration direction, which brings Ndelta in close proximity to the catalytic zinc. When the histidine becomes protonated, it then rotates to an "out" orientation, creating a more favorable solvation environment for the protonated His64. In this "out" orientation, the imidazole ring releases the delta nitrogen's excess proton into the bulk environment. After the second PT event and when the zinc-bound water is regenerated, the His64 is again favored to reorient to the "in" orientation, completing the catalytic cycle.     

The Right to Survive in North America

Surviving as Indians: The Challenge of Self-Government . Menno Boldt.
"Retained by the People": A History of American Indians and the Bill of Rights . John R. Wunder.     

Archaeal JAB1/MPN/MOV34 metalloenzyme (HvJAMM1) cleaves ubiquitin‐like small archaeal modifier proteins (SAMPs) from protein‐conjugates

Proteins with JAB1/MPN/MOV34 metalloenzyme (JAMM/MPN+) domains are widespread among all domains of life, yet poorly understood. Here we report the purification and characterization of an archaeal JAMM/MPN+ domain protein (HvJAMM1) from Haloferax volcanii that cleaves ubiquitin‐like small archaeal modifier proteins (SAMP1/2) from protein conjugates. HvJAMM1 cleaved SAMP1/2 conjugates generated in H. volcanii as well as isopeptide‐ and linear‐linked SAMP1–MoaE in purified form. Cleavage of linear linked SAMP1–MoaE was dependent on the presence of the SAMP domain and the C‐terminal VSGG motif of this domain. While HvJAMM1 was inhibited by size exclusion chromatography and metal chelators, its activity could be restored by addition of excess ZnCl₂. HvJAMM1 residues (Glu31, His88, His90, Ser98 and Asp101) that were conserved with the JAMM/MPN+ active‐site motif were required for enzyme activity. Together, these results provide the first example of a JAMM/MPN+ zinc metalloprotease that independently catalyses the cleavage of ubiquitin‐like (isopeptide and linear) bonds from target proteins. In archaea, HvJAMM1 likely regulates sampylation and the pools of ‘free’ SAMP available for protein modification. HvJAMM1‐type proteins are thought to release the SAMPs from proteins modified post‐translationally as well as those synthesized as domain fusions.     

The case of curers, noncurers, and biomedical experts in Pichátaro, Mexico. Resiliency in folk-medical beliefs

We explore potential conceptual and cultural change in folk-medical models within a Mexican community that may have taken place over the past 30 years. Building on a study from the 1970s, we explore the effects a government-supported biomedical clinic had on the content and distribution of folk-medical concepts. Surprisingly, we find that despite a dramatic increase in access to biomedicine and a host of socioeconomic shifts opening access to new medical ideas, folk-medical knowledge in Pichátaro, Michoacán, Mexico has remained largely unchanged with respect to its distribution and content. Curers and noncurers not only agree with one another but also continue to agree with a general model held in the 1970s. It is the medical models of clinic personnel that stand out as odd within the community. Yet, despite these conceptual differences, the biomedical facilities of the town are well attended.     

Development of breeder-friendly markers for selection of <Emphasis Type="Italic">MIPS1</Emphasis> mutations in soybean

Mutations in the d-myo-inositol 3-phosphate synthase 1 gene (MIPS1) in soybean [Glycine max (L.) Merr.] cause modifications to seed phosphorus and carbohydrate content that improve the nutritional value of food and feed. Molecular markers are an efficient tool for breeding MIPS1 mutant germplasm due to reduced seed germination and field emergence potential. An F₂ population segregating for the MIPS1 mutation found in experimental soybean line V99-5089 was used to develop breeder-friendly markers. Markers were validated in 88 advanced lines from 9 diverse pedigrees. Ten potential simple sequence repeat (SSR) markers, located on Gm11, from the new BARCSOYSSR_1.0 database were tested and four were polymorphic. BARCSOY_11_1495 was 93–97% effective for selecting the mutation. A KBiosciences Competitive Allele Specific PCR (KASPar) assay was developed to select directly for the V99-5089-derived MIPS1 single nucleotide polymorphism (SNP) mutation. The KASPar assay is simple and cost-effective compared to other SNP genotyping assays. The MIPS1 mutation in V99-5089 is likely to have occurred spontaneously. We describe a method of DNA extraction in soybean using a Geno/Grinder for fast and easy tissue maceration.     

Specific Glycoforms of MUC5AC and Endorepellin Accurately Distinguish Mucinous from Nonmucinous Pancreatic Cysts

Specific protein glycoforms may be uniquely informative about the pathological state of a cyst and may serve as accurate biomarkers. Here we tested that hypothesis using antibody-lectin sandwich arrays in broad screens of protein glycoforms and in targeted studies of candidate markers. We profiled 16 different glycoforms of proteins captured by 72 different antibodies in cyst fluid from mucinous and nonmucinous cysts (n = 22), and we then tested a three-marker panel in 22 addition samples and 22 blinded samples. Glycan alterations were not widespread among the proteins and were mainly confined to MUC5AC and endorepellin. Specific glycoforms of these proteins, defined by reactivity with wheat germ agglutinin and a blood group H antibody, were significantly elevated in mucinous cysts, whereas the core protein levels were not significantly elevated. A three-marker panel based on these glycoforms distinguished mucinous from nonmucinous cysts with 93% accuracy (89% sensitivity, 100% specificity) in a prevalidation sample set (n = 44) and with 91% accuracy (87% sensitivity, 100% specificity) in independent, blinded samples (n = 22). Targeted lectin measurements and mass spectrometry analyses indicated that the higher wheat germ agglutinin and blood group H reactivity was due to oligosaccharides terminating in GlcNAc or N-acetyl-lactosamine with occasional α1,2-linked fucose. The results show that MUC5AC and endorepellin glycoforms may be highly specific and sensitive biomarkers for the differentiation of mucinous from nonmucinous pancreatic cysts.Cysts in the pancreas can be a clinical challenge to patients and physicians, because some are precancerous and may progress to invasive cancer, whereas others remain indolent (1). The first step in diagnosis is to determine the type of cyst. Two types of pancreatic cysts—intraductal papillary mucinous neoplasms (IPMNs)¹ and mucinous cystic neoplasms (MCNs), together termed “mucinous cysts”—have malignant potential, and other types of cysts, such as serous cystadenomas (SCs) and pseudocysts (PCs), are essentially benign. To date, there are difficulties in correctly determining the cyst type. The methods commonly used to assess pancreatic cysts are endoscopic ultrasound and cyst fluid aspiration followed by the carcinoembryonic antigen (CEA) assay and cytology. The most useful is CEA, which distinguishes mucinous cysts from nonmucinous cysts with 70% to 80% accuracy (2–4). Cytology (microscopic analysis of the cells in the fluid) has limited value because of the lack of consistency in obtaining reliable cellular material (2). Other biomarkers, including DNA analysis, have yet to achieve widespread use (5).Because the current tests are not conclusive for many patients, new, more effective molecular markers are needed to provide more accurate diagnoses and to help guide management. Researchers have investigated molecular features such as DNA mutations (6, 7), specific protein levels (8), microRNAs (9), inflammatory cytokines (10), and the presence of mucin (11, 12). Broad searches utilizing genomic, proteomic, and glycomic profiling (13–15) have uncovered other candidate biomarkers that require further study and validation.We previously investigated whether specific protein glycoforms are present exclusively in the fluid from mucinous cysts (16). The reason for investigating specific glycoforms, rather than simply the protein levels, is that the glycoforms may be more closely associated with disease. Cells can rework the glycosylation machinery as they become neoplastic and progress to invasive cancer. For example, cancer cells and dysplastic cells can enhance N-glycan complexity through increased branching (17), produce truncated O-glycans through the lost activity of an enzyme critical for O-glycan extension (18), or increase fucosylation or the production of Lewis structures (19). Such glycan restructuring can affect cellular adhesiveness, migration, cytokine signaling, receptor recycling, or immune cell interactions (20), all potentially involved in cancer progression. Therefore, we potentially can get more information about the differentiation state of a cell by detecting protein glycosylation in addition to protein abundance.The protein MUC5AC in cyst fluid demonstrated such potential in a previous study by our group (16). We examined both protein levels and specific glycoform levels of MUC5AC in fluid from mucinous cysts relative to nonmucinous cysts. A glycoform of MUC5AC, one that binds the lectin wheat germ agglutinin (WGA), was almost exclusively found in the mucinous cysts and was a better biomarker than the MUC5AC protein measured over all glycoforms. Furthermore, the glycosylation patterns in pancreatic cysts were different from those we found in pancreatic ductal adenocarcinomas (21, 22).In the current study, we performed a broad search of addition glycoforms of MUC5AC and other proteins that may be up-regulated in mucinous cysts and tested the hypothesis that the detection of specific glycoforms can serve to accurately discriminate mucinous from nonmucinous cysts. We confirmed the importance of the WGA-reactive glycoform of MUC5AC and found a second glycoform of MUC5AC, displaying the blood group H (BGH) antigen, up-regulated in mucinous cysts. We found a new marker, endorepellin, that also displayed elevated WGA-reactive and BGH-reactive glycoforms in mucinous cysts. A panel of three markers, comprising two glycoforms of MUC5AC and one of endorepellin, classified the samples with high sensitivity and specificity, providing the potential for more accurate diagnoses of pancreatic cysts. We further characterized the diagnostic glycoforms using mass spectrometry and targeted lectin analyses, which provided information about the molecular features of cystic precursors of cancer and routes for the further development of biomarker assays.