Applications of a “Whole Community” Framework for Enhancing Community or Campus Resilience

AbstractThe Community and Regional Resilience Institute (CARRI) has developed a unique approach to community resilience based on a “Whole Community” concept. It treats communities as a collection of systems, each with its own resilience. CARRI has applied its approach to two kinds of communities: civil communities, and institutions of higher education (IHEs). For both civil communities and IHEs, CARRI carried out a pilot program. For each participant, their leadership directed an assessment of the resilience of the component systems to the types of changes most relevant to that community. Each assessment provided suggestions for filling any gaps identified as part of the assessment. The pilot for the seven IHEs followed that for the seven civil communities and was able to take advantage of lessons learned from the first. These two pilot programs led to the following conclusions:•CARRI's systems-based approach is both understandable and usable by both types of communities. In practice, it seemed to provide a natural way to look at a community.•In general, IHEs were able to make better use of the approach than civil communities. This is due, in part, to the improvements made in the IHE pilot program based on the civil communities’ results. However, it also reflects the more hierarchical nature of IHEs, the tighter coupling of systems within an IHE and greater discretion in the use of resources in an IHE.•College campuses can be crucial catalysts for enhancing the resilience of civil communities.•Leadership is a key, perhaps the key, element in the success of a community resilience initiative

gyrA mutations and phenotypic susceptibility levels to ofloxacin and moxifloxacin in clinical isolates of Mycobacterium tuberculosis

Objectives To compare mutations in the quinolone resistance-determining region of the gyrA gene and flanking sequences with the MICs of ofloxacin and moxifloxacin for Mycobacterium tuberculosis. Methods The presence of mutations in 177 drug-resistant M. tuberculosis isolates was determined by DNA sequencing and the MICs quantified by MGIT 960. Results Single nucleotide polymorphisms were detected at codons 94 (n = 30), 90 (n = 12), 91 (n = 3), 89 (n = 1), 88 (n = 1) and 80 (n = 1). Four isolates with double mutations D94G plus A90V (n = 2) and D94G plus D94N (n = 2) reflect mixed populations. Agreement between genotypic and phenotypic susceptibility was high (≥97%) for both drugs. Mutant isolates had an MIC50 of 8.0 mg/L and an MIC90 of >10 mg/L for ofloxacin compared with an MIC50 and MIC90 of 2.0 mg/L for moxifloxacin. Codons 94 and 88 were linked to higher levels of fluoroquinolone resistance compared with codons 90, 91 and 89. The MIC distributions for the wild-type isolates ranged from ≤0.5 to 2.0 mg/L for ofloxacin and from ≤0.125 to 0.25 mg/L for moxifloxacin. However, 96% of the isolates with genetic alterations had MICs ≤2.0 mg/L for moxifloxacin, which is within its achievable serum levels. Conclusions This study provides quantitative evidence that the addition of moxifloxacin to extensively drug-resistant tuberculosis (XDR-TB) regimens based on a clinical breakpoint of 2.0 mg/L has merit. The use of moxifloxacin in the treatment of multidrug-resistant tuberculosis may prevent the acquisition of additional mutations and development of XDR-T

Genetic Susceptibility of Cultured Shrimp (\u3ci\u3ePenaeus vannamei\u3c/i\u3e) to Infectious Hypodermal and Hematopoietic Necrosis Virus and \u3ci\u3eBaculovirus penaei\u3c/i\u3e: Possible Relationship with Growth Status and Metabolic Gene Expression

Offspring of four crosses (I, II, III, and IV) of Penaeus vannamei from known high- and low-growth families were challenged with infectious hypodermal and hematopoietic necrosis virus (IHHNV) and Baculovirus penaei (BP) to compare their susceptibility to these viral agents and examine the genetic component involved in disease resistance or susceptibility. Family crosses were made using broodstock from five families developed by the U.S. Marine Shrimp Farming Program. The prevalence of IHHNV infection was highest in cross I and lowest in cross III. Cross I was developed using male and female broodstock from the low-growth family 1.6, and cross III was developed using a female from the high-growth family 1.3 and a male from the low-growth family 1.6. The prevalence of BP infection at Day 4 was highest (100%) in cross IV, which was developed using a female from the low-growth family 1.4 and a male from the high-growth family 1.5. The reciprocal cross, cross III, had the lowest (68%) prevalence at Day 4 postexposure. Both crosses I and II had 88% prevalence of infection at Day 4. Despite 100% prevalence of BP infection in cross IV at 4 days, animals from this cross and cross II exhibited high survival by Day 18 (85 and 77%). On the other hand, crosses I and III (with 88 and 68% prevalence at Day 4, respectively) showed low survival at Day 18 (19 and 24%). On the basis of prevalence of infection and mortality rates, it was concluded that the susceptibility to BP in penaeid shrimp is governed by the genetic background of the parental crosses. The random amplified polymorphic DNA polymorphisms for crosses I, II, III, and IV, were 43, 45, 53, and 51%, respectively, showing no clear relationship between IHHNV and BP prevalence of infection and levels of nuclear genetic diversity. Though the mtDNA haplotypes in offspring from the different crosses were the same, major differences were observed in both steady-state levels and patterns of expression of the mitochondrial 12s rRNA in offspring obtained at various early developmental stages from each of the four crosses. The possible relationship among disease susceptibility, growth status, and expression of mitochondrial 12s rRNA is discussed in the context of a complex nuclear-cytoplasmic genetic system involved in the regulation of gene expression

The actin-binding protein Hip1R associates with clathrin during early stages of endocytosis and promotes clathrin assembly in vitro

Huntingtin-interacting protein 1 related (Hip1R) is a novel component of clathrin-coated pits and vesicles and is a mammalian homologue of Sla2p, an actin-binding protein important for both actin organization and endocytosis in yeast. Here, we demonstrate that Hip1R binds via its putative central coiled-coil domain to clathrin, and provide evidence that Hip1R and clathrin are associated in vivo at sites of endocytosis. First, real-time analysis of Hip1R–YFP and DsRed–clathrin light chain (LC) in live cells revealed that these proteins show almost identical temporal and spatial regulation at the cell cortex. Second, at the ultrastructure level, immunogold labeling of ‘unroofed’ cells showed that Hip1R localizes to clathrin-coated pits. Third, overexpression of Hip1R affected the subcellular distribution of clathrin LC. Consistent with a functional role for Hip1R in endocytosis, we also demonstrated that it promotes clathrin cage assembly in vitro. Finally, we showed that Hip1R is a rod-shaped apparent dimer with globular heads at either end, and that it can assemble clathrin-coated vesicles and F-actin into higher order structures. In total, Hip1R's properties suggest an early endocytic function at the interface between clathrin, F-actin, and lipids

Evolution and expansion of the Mycobacterium tuberculosis PE and PPE multigene families and their association with the duplication of the ESAT-6 (esx) gene cluster regions

BACKGROUND: The PE and PPE multigene families of Mycobacterium tuberculosis comprise about 10% of the coding potential of the genome. The function of the proteins encoded by these large gene families remains unknown, although they have been proposed to be involved in antigenic variation and disease pathogenesis. Interestingly, some members of the PE and PPE families are associated with the ESAT-6 (esx) gene cluster regions, which are regions of immunopathogenic importance, and encode a system dedicated to the secretion of members of the potent T-cell antigen ESAT-6 family. This study investigates the duplication characteristics of the PE and PPE gene families and their association with the ESAT-6 gene clusters, using a combination of phylogenetic analyses, DNA hybridization, and comparative genomics, in order to gain insight into their evolutionary history and distribution in the genus Mycobacterium. RESULTS: The results showed that the expansion of the PE and PPE gene families is linked to the duplications of the ESAT-6 gene clusters, and that members situated in and associated with the clusters represent the most ancestral copies of the two gene families. Furthermore, the emergence of the repeat protein PGRS and MPTR subfamilies is a recent evolutionary event, occurring at defined branching points in the evolution of the genus Mycobacterium. These gene subfamilies are thus present in multiple copies only in the members of the M. tuberculosis complex and close relatives. The study provides a complete analysis of all the PE and PPE genes found in the sequenced genomes of members of the genus Mycobacterium such as M. smegmatis, M. avium paratuberculosis, M. leprae, M. ulcerans, and M. tuberculosis. CONCLUSION: This work provides insight into the evolutionary history for the PE and PPE gene families of the mycobacteria, linking the expansion of these families to the duplications of the ESAT-6 (esx) gene cluster regions, and showing that they are composed of subgroups with distinct evolutionary (and possibly functional) differences

Prevalence of pyrazinamide resistance across the spectrum of drug resistant phenotypes of Mycobacterium tuberculosis

Pyrazinamide resistance is largely unknown in the spectrum of drug resistant phenotypes. We summarize data on PZA resistance in clinical isolates from South Africa. PZA DST should be performed when considering its inclusion in treatment of patients with rifampicin-resistant TB or MDR-TB

A Combined Phenotypic-Genotypic Predictive Algorithm for In Vitro Detection of Bicarbonate: β-Lactam Sensitization among Methicillin-Resistant Staphylococcus aureus (MRSA).

Antimicrobial susceptibility testing (AST) is routinely used to establish predictive antibiotic resistance metrics to guide the treatment of bacterial pathogens. Recently, a novel phenotype termed "bicarbonate (NaHCO3)-responsiveness" was identified in a relatively high frequency of clinical MRSA strains, wherein isolates demonstrate in vitro "susceptibility" to standard β-lactams (oxacillin [OXA]; cefazolin [CFZ]) in the presence of NaHCO3, and in vivo susceptibility to these β-lactams in experimental endocarditis models. We investigated whether a targeted phenotypic-genotypic screening of MRSA could rule in or rule out NaHCO3 susceptibility upfront. We studied 30 well-characterized clinical MRSA bloodstream isolates, including 15 MIC-susceptible to CFZ and OXA in NaHCO3-supplemented Mueller-Hinton Broth (MHB); and 15 MIC-resistant to both β-lactams in this media. Using a two-tiered strategy, isolates were first screened by standard disk diffusion for susceptibility to a combination of amoxicillin-clavulanate [AMC]. Isolates then underwent genomic sequence typing: MLST (clonal complex [CC]); agr; SCCmec; and mecA promoter and coding region. The combination of AMC disk susceptibility testing plus mecA and spa genotyping was able to predict MRSA strains that were more or less likely to be NaHCO3-responsive in vitro, with a high degree of sensitivity and specificity. Validation of this screening algorithm was performed in six strains from the overall cohort using an ex vivo model of endocarditis. This ex vivo model recapitulated the in vitro predictions of NaHCO3-responsiveness vs. nonresponsiveness above in five of the six strains

Detection of natural infection with Mycobacterium intracellulare in healthy wild-caught Chacma baboons (Papio ursinus) by ESAT-6 and CFP-10 IFN-γ ELISPOT tests following a tuberculosis outbreak

<p>Abstract</p> <p>Background</p> <p>Both tuberculous and non-tuberculous mycobacteria can cause infection in nonhuman primates (NHP), indicating the existence of potential zoonotic transmission between these animals and visitors to zoos or animal handlers in primate facilities. Screening of mycobacterial infections in NHP is traditionally done by tuberculin skin test (TST), which is unable to distinguish between pathogenic and non-pathogenic mycobacterial infections. In this study, we investigated the use of ESAT-6 and CFP-10 for detection of mycobacterial infections in a wild-caught baboon colony after one baboon died of tuberculosis (TB).</p> <p>Methods</p> <p>Peripheral blood lymphocytes for interferon-gamma enzyme-linked immunospot assay (IFN-γ ELISPOT) assay were obtained from TST positive baboons and those in contact with tuberculous baboons before being euthanased, autopsied and lung tissues taken for histology and mycobacterial culture.</p> <p>Results</p> <p>Both ESAT-6 and CFP-10 IFN-γ ELISPOT assays were able to detect early <it>M. tuberculosis </it>but also <it>M. intracellulare </it>infection. Although this indicates potential cross-reactivity with <it>M. intracellulare </it>antigens, the method was able to distinguish <it>M. bovis </it>BCG vaccination from <it>M. tuberculosis </it>infection. This assay performed better than the TST, which failed to detect one <it>M. tuberculosis </it>and two early <it>M. intracellulare </it>infections.</p> <p>Conclusion</p> <p>These results suggest that the IFN-γ ELISPOT assay could improve the detection of <it>M tuberculosis </it>infections when screening NHP. There is some doubt, however, concerning specificity, as the assay scored positive three animals infected with <it>M. intracellulare</it>.</p