Coupling of binding and differential subdomain folding of the intrinsically disordered transcription factor CREB.

The cyclic AMP response element binding protein (CREB) contains a basic leucine zipper motif (bZIP) that forms a coiled coil structure upon dimerization and specific DNA binding. Although this state is well characterized, key features of CREB bZIP binding and folding are not well understood. We used single-molecule Förster resonance energy transfer (smFRET) to probe conformations of CREB bZIP subdomains.

The material properties of a bacterial-derived biomolecular condensate tune biological function in natural and synthetic systems.

Intracellular phase separation is emerging as a universal principle for organizing biochemical reactions in time and space. It remains incompletely resolved how biological function is encoded in these assemblies and whether this depends on their material state. The conserved intrinsically disordered protein PopZ forms condensates at the poles of the bacterium Caulobacter crescentus, which in turn orchestrate cell-cycle regulating signaling cascades.

Conformational Freedom and Topological Confinement of Proteins in Biomolecular Condensates.

The emergence of biomolecular condensation and liquid-liquid phase separation (LLPS) introduces a new layer of complexity into our understanding of cell and molecular biology. Evidence steadily grows indicating that condensates are not only implicated in physiology but also human disease. Macro- and mesoscale characterization of condensates as a whole have been instrumental in understanding their biological functions and dysfunctions.

A topological switch in CFTR modulates channel activity and sensitivity to unfolding.

The cystic fibrosis transmembrane conductance regulator (CFTR) anion channel is essential to maintain fluid homeostasis in key organs. Functional impairment of CFTR due to mutations in the cftr gene leads to cystic fibrosis. Here, we show that the first nucleotide-binding domain (NBD1) of CFTR can spontaneously adopt an alternate conformation that departs from the canonical NBD fold previously observed.

Micromotion Analysis of Various Tibial Constructs in Moderate Tibial Defects in Revision Total Knee Arthroplasty.

Background: The purpose of this study is to compare the micromotion of various tibial reconstruction strategies including short cemented and long cementless stems with or without metaphyseal augmentation.

Methods: A moderate tibial bone defect was milled into dual density polyurethane test blocks. Mechanical testing was performed on 4 test constructs: (1) short cemented stem (75-mm total length) alone; (2) short cemented stem with a symmetric metaphyseal cone; (3) a press-fit (175-mm total length) diaphyseal engaging tibial construct without a cone, and (4) the same press-fit tibial construct with a metaphyseal cone augment.

A nucleotide-switch mechanism mediates opposing catalytic activities of Rel enzymes.

Bifunctional Rel stringent factors, the most abundant class of RelA/SpoT homologs, are ribosome-associated enzymes that transfer a pyrophosphate from ATP onto the 3' of guanosine tri-/diphosphate (GTP/GDP) to synthesize the bacterial alarmone (p)ppGpp, and also catalyze the 3' pyrophosphate hydrolysis to degrade it. The regulation of the opposing activities of Rel enzymes is a complex allosteric mechanism that remains an active research topic despite decades of research. We show that a guanine-nucleotide-switch mechanism controls catalysis by Thermus thermophilus Rel (Rel).

Domain-interface dynamics of CFTR revealed by stabilizing nanobodies.

The leading cause of cystic fibrosis (CF) is the deletion of phenylalanine 508 (F508del) in the first nucleotide-binding domain (NBD1) of the cystic fibrosis transmembrane conductance regulator (CFTR). The mutation affects the thermodynamic stability of the domain and the integrity of the interface between NBD1 and the transmembrane domain leading to its clearance by the quality control system. Here, we develop nanobodies targeting NBD1 of human CFTR and demonstrate their ability to stabilize both isolated NBD1 and full-length protein.

Diagnosing intramammary infection: Controlling misclassification bias in longitudinal udder health studies.

Using imperfect tests may lead to biased estimates of disease frequency and of associations between risk factors and disease. For instance in longitudinal udder health studies, both quarters at risk and incident intramammary infections (IMI) can be wrongly identified, resulting in selection and misclassification bias, respectively. Diagnostic accuracy can possibly be improved by using duplicate or triplicate samples for identifying quarters at risk and, subsequently, incident IMI.

Tibial shaft anatomy differs between Caucasians and East Asian individuals.

Purpose: The orientation and distance from the shaft of the femur and tibia to the articular surface centre is important for performing total knee arthroplasty (TKA) with cementless stems. It is important to understand anatomic differences between races to match the tibial and femoral shaft axis to the knee articular surface. Thus, the purpose of this study was to compare knee morphology between Caucasian and East Asian individuals to determine the optimal placement of tibial and femoral stems.

Draft Genome Sequence of Bovine Mastitis Isolate Staphylococcus agnetis CBMRN 20813338.

Presented here is a draft genome sequence for Staphylococcus agnetis CBMRN 20813338, isolated from a lactating dairy cow with subclinical mastitis. The genome is approximately 2,416 kb and has 35.79% G+C content.

Benchmarks for evaluation and comparison of udder health status using monthly individual somatic cell count.

The objectives of this study were to propose benchmarks for the interpretation of herd udder health using monthly individual somatic cell counts (SCC) from dairy herds in Quebec, Canada and to evaluate the association of risk factors with intramammary infection (IMI) dynamics relative to these benchmarks. The mean and percentiles of indices related to udder infection status [e.g.

Efficacy of extended intramammary ceftiofur therapy against mild to moderate clinical mastitis in Holstein dairy cows: a randomized clinical trial.

Few studies have investigated the efficacy of extended ceftiofur therapy and none have focused on extended therapy for naturally occurring clinical mastitis. The objective of this study was to compare the efficacy of extended intramammary ceftiofur therapy of 8 d duration with a standard 2-day regimen for the treatment of naturally occurring mild to moderate clinical mastitis in lactating dairy cows. Holstein cows from 22 dairy herds (n = 241) were randomly allocated to the 2 treatment groups.

Herd-level relationship between antimicrobial use and presence or absence of antimicrobial resistance in gram-negative bovine mastitis pathogens on Canadian dairy farms.

Concurrent data on antimicrobial use (AMU) and resistance are needed to contain antimicrobial resistance (AMR) in bacteria. The present study examined a herd-level association between AMU and AMR in Escherichia coli (n=394) and Klebsiella species (n=139) isolated from bovine intramammary infections and mastitis cases on 89 dairy farms in 4 regions of Canada [Alberta, Ontario, Québec, and Maritime Provinces (Prince Edward Island, Nova Scotia, and New Brunswick)]. Antimicrobial use data were collected using inventory of empty antimicrobial containers and antimicrobial drug use rate was calculated to quantify herd-level AMU.

The expression of a putative exotoxin and an ABC transporter during bovine intramammary infection contributes to the virulence of Staphylococcus aureus.

Staphylococcus aureus is a leading cause of intramammary infections (IMI) and bovine mastitis is an important disease for the dairy industry. As this bacterium probably expresses specific genes for establishment of IMI, we studied the transcriptional profile of four S. aureus strains recovered from experimentally infected cows.

Management practices associated with the bulk-milk prevalence of Staphylococcus aureus in Canadian dairy farms.

When designing mastitis-prevention and control programs, it is important to know the level of adoption of mastitis-prevention management practices and control programs and the herd-level prevalence of contagious mastitis pathogens. Our objectives were to estimate: (1) adoption of recommended mastitis-preventive management on Canadian dairy farms; (2) herd-level prevalence of contagious mastitis pathogens on Canadian dairy farms; and (3) associations of certain management practices with the isolation of Staphylococcus aureus from the bulk tank milk from Canadian dairy farms. In total, 226 farms participating in dairy herd improvement milk recording were randomly selected.

Development and validation of a bilingual questionnaire for measuring udder health related management practices on dairy farms.

Questionnaires are frequently used instruments to collect data in epidemiological studies. In countries where more than one language is spoken, the development of a questionnaire in more than one language is needed. The objective of this study was to develop and test the repeatability and validity of English and French versions of a personal interview-format questionnaire designed to capture udder health related management practices used on dairy farms.

Evaluation of the California Mastitis Test as a precalving treatment selection tool for Holstein heifers.

The objective of this study was to evaluate the California Mastitis Test (CMT) and a portable electrical conductivity meter for diagnosing precalving intramammary infection (IMI) in Holstein heifers. A total of 428 dairy heifers from 23 dairy herds were enrolled between 6 and 12 days before the expected calving date from June 2002 to June 2003. Mammary secretions were tested by both diagnostic methods and by bacterial culture for evidence of IMI.

Effect of precalving intramammary treatment with pirlimycin in nulliparous Holstein heifers.

A clinical trial was conducted to determine whether prepartum intramammary pirlimycin reduces the proportion of nulliparous heifers with intramammary infection (IMI) during early lactation and improves milk production. Quarter milk samples were collected from 428 heifers, systematically allocated to treatment and control groups, 6 to 12 d before the expected calving date and 2 to 8 d after calving. At the prepartum visit, heifers in the treatment group (n = 219) received an infusion of pirlimycin hydrochloride in all 4 quarters; the control heifers (n = 209) received no infusions.

Climatic and demographic determinants of American visceral leishmaniasis in northeastern Brazil using remote sensing technology for environmental categorization of rain and region influences on leishmaniasis.

Remote sensing (RS) permits evaluation of spatial and temporal variables that can be used for vector-borne disease models. A Landsat Thematic Mapper scene covering Canindé, Ceará in northeastern Brazil (September 25, 1986) was spectrally enhanced and classified using ERDAS (Atlanta, GA) Imagine for 873 4-km2 areas. The population and number of cases of American visceral leishmaniasis (AVL) were determined for each 4-km2 area.

Prevalence and incidence density of Mycobacterium leprae and Trypanosoma cruzi infections within a population of wild nine-banded armadillos.

A total of 415 wild 9-banded armadillos from the East Atchafalaya River Levee (Point Coupee, LA) were collected over 4 years to estimate the incidence and prevalence of Mycobacterium leprae and Trypanosoma cruzi and to discern any relationship between the 2 agents. M. leprae infections were maintained at a high steady prevalence rate year to year averaging 19%.

Screening for syphilis in arrestees: usefulness for community-wide syphilis surveillance and control.

Background: Syphilis screening of jail arrestees has been promoted as an effective method for both disease control and surveillance.

Goals: To evaluate the yield of the East Baton Rouge Parish Jail screening program in detecting previously undiagnosed syphilis, to evaluate the program as a means for monitoring community syphilis rates, and to characterize arrestees at greatest risk for syphilis infection.

Study Design: From July 1994 to December 1998, arrestees brought to the East Baton Rouge Parish Jail were screened for syphilis.

Clinical hematology practices at veterinary teaching hospitals and private diagnostic laboratories.

The clinical hematology practices utilized at veterinary teaching hospitals and private veterinary diagnostic laboratories were surveyed using a questionnaire. The hematology caseload at private diagnostic laboratories was larger, and comprised predominantly of canine and feline submissions. The Coulter S Plus IV and Serono Baker 9000 were the hematology analyzers used most frequently at veterinary medical laboratories.