Interpretation and reporting considerations of low-level DNA profiles following MinElute® purification of PowerPlex® 21 amplified products.

The generation of high-quality DNA profiles from trace amounts of DNA continues to be an issue in forensic casework. Several methods have been proposed over the years to increase recovery rates for low input DNA, including purification of PCR products, an increase in PCR cycle numbers and increasing injection time or voltage during electrophoresis. In this study, the characteristics of DNA profiles generated using QIAGEN MinElute® purification of Promega PowerPlex® 21 amplified products for low DNA input samples, ranging from 80 pg down to 4 pg, were evaluated. MinElute® purification was found to be a simple, effective and time efficient method, which can greatly improve the resolution of amplified PCR products, recovering 100% of donor concordant alleles from as little 16 pg of input template DNA and generating sufficient allelic information for interpretation from as low as 4 pg inputs. However, as is commonly observed with low template DNA samples, the results exhibited extensive disparity in the effects of stochastic variation in amplification, including increased heterozygote peak height imbalance, stutter ratios and instances of allelic drop-in and drop-out, both within and between replicates. As such, it is important that the extent and variability of these stochastic effects are appropriately incorporated in the development of robust profile interpretation guidelines for DNA profiles generated from purified PCR products.

Telemedicine and Its Perceptions in a Border Community: A Review of How Health Care Technology Has Helped Increase Access.

Introduction: Telemedicine is a practical way of offering medical services to remote and underserved areas. During the COVID-19 pandemic, telemedicine has provided convenient access to health care and has overcome barriers such as distance that prevent patients from receiving care. Border populations are impacted by this change in health care delivery. The goal of this study was to investigate how a border patient population perceives their experiences with telemedicine. Methods: We utilized telephone surveys of patients who had a recent telehealth visit at the Texas Tech University Health Science Center (TTUHSC) Family Medicine Center clinic in El Paso, Texas. Survey measures included patients' demographics, a quality assessment of the patients' most recent telehealth visit and their experience, a comparison of the patients' telehealth visit to past in-person visits, and a rating of their telehealth visit. Result: Over 2,000 individuals (n = 2,040), primarily Hispanic females, older than the age of 44 years were identified for potential inclusion in the study. Of these, 928 had a contact attempt, of which 1,378 could not be contacted, 592 were invited, 70 declined leading to a response rate of 67.6% (number invited/completed the survey). Most patients agreed that during their most recent telehealth visit their clinician listened well (98.7%), spent adequate time with them (98.2%), was prompt (94.5%), explained things well (98.0%), and was someone they would recommend to others (97.2%). When comparing telehealth to in-person visits, patients reported the following: less wait time, easier convenience, and similar quality between virtual and in-person visits. Patients rated both their likelihood of using telehealth again and their likelihood of recommending telehealth to others as an 8.68 out of 10, on average. Patients 65 years old or older had 3.17 times greater likelihood of satisfaction with virtual visits when compared with patients younger than 45 years old (confidence interval [95% CI], 1.24-11.11). Patients also had less satisfaction with virtual visits if they had lower educational attainment (odds ratio = 0.10; 95% CI, 0.01-0.81). Conclusions: We found that individuals in a border community had a positive experience with telehealth primary care visits. This approach may improve access to health care.

A comparison of six adhesive tapes as tape lifts for efficient trace DNA recovery without the transfer of PCR inhibitors.

Tape-lifting is a non-destructive method employed in the laboratory to recover and collect trace DNA evidence from crime scene exhibits with porous surfaces. The success of tape-lifting is a balance between capturing the biological material and compatibility with downstream DNA extraction processes to ensure efficient release of the tape-lifted material during DNA extraction. In this study, six commercially available low-, regular- and high-tack adhesive tapes were evaluated. The low-tack S183 tape and the highly adhesive S-Hold tape were compared for DNA recovery efficiency from different materials commonly encountered in casework. All tape-lifts were processed using PrepFiler Express™ BTA and AutoMate Express™ Forensic DNA extraction systems, DNA samples quantitated by Quantifiler TRIO, amplified using Powerplex® 21 and VeriFiler™ PLUS (VFP), and analysed on a 3500xl genetic analyser to evaluate the quality of the resultant STR profiles obtained. The more adhesive S-Hold tape recovered comparable or more DNA than the low-tack S183 tape from the majority of materials tested. However, STR profiles obtained from S183 tape-lifts were of markedly higher quality compared to S-Hold tape-lifts. This was most evident for towel, denim and printed chiffon, where S-Hold samples exhibited severe PCR inhibition, with VFP internal quality markers confirming the presence of inhibitors. The findings suggest that strong adhesion is not necessarily beneficial for tape-lifting, as the low tack S183 tape was able to efficiently recover cellular material from the surface of porous substrates commonly encountered in casework, while avoiding the co-transfer of PCR-inhibitory substances from the sampled material.

The importance of transects for characterizing aged organic contaminant plumes in groundwater.

A complex mixture of dissolved organic contaminants, emanating from a many decades-old, residual, dense non-aqueous phase liquid (DNAPL) source, migrates through unconfined, moderately heterogeneous, glacial-derived sediments and sedimentary rock in a residential area of Dane County, Wisconsin, USA. A portion of this contaminant plume intersects a large man-made pond, roughly 400 m downgradient of the source zone. Depth-discrete, multilevel groundwater sampling, detailed sedimentological logs, and hydraulic head profiles were used to delineate the spatial distribution of hydraulic, geologic, organic contaminant, and redox hydrochemical conditions within the established plume along two transects immediately upgradient of the pond. Twenty-one contaminants were detected and classified into four major contaminant groups: chlorinated ethenes, chlorinated ethanes, aromatics (BTEX: benzene, toluene, ethylbenzene, xylene), and aliphatic ketones. Within the glacial sediments and shallow bedrock, zones of reductive dechlorination of chlorinated ethenes and ethanes were juxtaposed with zones of BTEX and ketone degradation. Spatial heterogeneity in the concentration and distribution of contaminant groups and redox conditions was observed over lateral distances of tens of meters and vertical distances of tens of centimeters along the two transects. Although the site was situated in a complex glacial depositional environment, lithologic and hydraulic heterogeneity surprisingly only had a modest influence on the spatial distribution of plume contaminants. Depth-discrete sampling along paired, closely spaced transects (~20 m apart) was essential to assess internal plume composition/concentration evolution along flow paths with strong attenuation over short migration distances. This study shows how paired, highly resolved transects can enhance understanding of transverse and longitudinal variability in areas where contaminant-induced redox conditions control reaction zones and strong plume attenuation.

Improving Youth Suicide Risk Screening and Assessment in a Pediatric Hospital Setting by Using The Joint Commission Guidelines.

OBJECTIVES: Hospitals accredited by The Joint Commission (TJC) are now required to use a validated screening tool and a standardized method for assessment of suicide risk in all behavioral health patients. Our aims for this study were (1) to implement a TJC-compliant process of suicide risk screening and assessment in the pediatric emergency department (ED) and outpatient behavioral health clinic in a large tertiary care children's hospital, (2) to describe characteristics of this population related to suicide risk, and (3) to report the impact of this new process on ED length of stay (LOS). METHODS: A workflow using the Columbia Suicide Severity Rating Scale was developed and implemented. Monthly reviews of compliance with screening and assessment were conducted. Descriptive statistics were used to define the study population, and multivariable regression was used to model factors associated with high suicide risk and discharge from the ED. ED LOS of behavioral health patients was compared before and after implementation. RESULTS: Average compliance rates for screening was 83% in the ED and 65% in the outpatient clinics. Compliance with standardized assessments in the ED went from 0% before implementation to 88% after implementation. The analysis revealed that 72% of behavioral health patients in the ED and 18% of patients in behavioral health outpatient clinics had a positive suicide risk. ED LOS did not increase. The majority of patients screening at risk was discharged from the hospital after assessment. CONCLUSIONS: A TJC-compliant process for suicide risk screening and assessment was implemented in the ED and outpatient behavioral health clinic for behavioral health patients without increasing ED LOS.

The use of bacteriophage MS2 for the development and application of a virucide decontamination test method for porous and heavily soiled surfaces.

AIMS: (i) To develop an analytical method for recovery and quantification of bacteriophage MS2-as a surrogate for foot-and-mouth disease virus-from complex porous surfaces, with and without the presence of laboratory-developed agricultural grime; (ii) to evaluate, with a 4-log dynamic range, the virucidal activity of common biocides for their ability to decontaminate surfaces and hence remediate facilities, following a foreign animal disease contamination incident. METHODS AND RESULTS: An analytical method was developed and optimized for MS2 recovery from simulated agricultural surfaces. The addition of Dey-Engley neutralizing broth to an extraction buffer improved MS2 viability in liquid extracts, with optimal analytical holding times determined as <8 to ≤24 h, depending on matrix. The recovery of MS2 from surface materials decreased in the order: nonporous reference material >grimed porous materials >nongrimed porous materials. In disinfectant testing, two spray applications of pAB were effective against MS2 (≥4-log reduction) on all operational-scale materials. Two per cent citric acid had limited effectiveness, with a ≥4-log reduction observed on a selected subset of grimed concrete samples. CONCLUSIONS: Decontamination efficacy test results can be affected by surface characteristics, extraction buffer composition, analytical holding time and surface-specific organism survivability. Efficacy should be evaluated using a test method that reflects the environmental characteristics of the intended application. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this study demonstrate the importance of analytical method verification tests for disinfectant testing prior to application in complex environments.

Methane gas transport in unconfined aquifers: A numerical sensitivity study of a controlled release experiment at CFB Borden.

Subsurface leakage of natural gas from petroleum wells can impact freshwater aquifers. Accurate prediction of gas migration in the subsurface will depend on knowledge of permeability, porosity, and flow system conditions. A series of two-dimensional numerical multi-phase flow simulations (CFbio) were conducted to investigate the role of multi-phase parameters (relative permeability and air entry pressure), flow system conditions (intrinsic permeability, anisotropy, and groundwater velocity), and geometric properties (layer thickness and layer lateral continuity) on the flow of gas-phase methane emanating from two variable-rate point sources in an unconfined sandy aquifer. Numerical simulations showed that for a homogeneous, weakly anisotropic aquifer, gas migrates almost exclusively vertically due to buoyancy, before venting to the vadose zone and atmosphere. As vertical migration became restricted through increased anisotropy, inclusion of lower-permeable layers, or increased horizontal groundwater velocity, an increase in the lateral component of gas migration was observed. This led to the formation of a broader lateral migration of the gas-phase plume and establishment of variably distributed vertical preferential flow paths, ultimately resulting in increased gas retention in the aquifer with relatively less methane reaching the vadose zone or atmosphere. The inclusion of a thin layer with moderately lower permeability (1-2 orders of magnitude) and increased air entry pressure was used to depict a fine-grained sand lens within a uniform aquifer. This subtle feature led to the formation of thin gas pools extending up- and down-gradient beneath the lens, allowing methane to travel much farther and faster than by groundwater advection alone, which is consistent with field observations during the experiment. In all scenarios investigated gas-phase methane was shown to migrate predominantly vertically due to buoyancy, until the aquitard permeability was <30% of the aquifer permeability. Our modelling demonstrates that even subtle permeability contrasts, together with capillary pressure changes demarcating grain-scale bedding, will lead to extensive lateral free-phase gas migration, and development of a more extensive and complex zone of impacted aquifer than presupposed.

Campylobacter jejuni Demonstrates Conserved Proteomic and Transcriptomic Responses When Co-cultured With Human INT 407 and Caco-2 Epithelial Cells.

Major foodborne bacterial pathogens, such as Campylobacter jejuni, have devised complex strategies to establish and foster intestinal infections. For more than two decades, researchers have used immortalized cell lines derived from human intestinal tissue to dissect C. jejuni-host cell interactions. Known from these studies is that C. jejuni virulence is multifactorial, requiring a coordinated response to produce virulence factors that facilitate host cell interactions. This study was initiated to identify C. jejuni proteins that contribute to adaptation to the host cell environment and cellular invasion. We demonstrated that C. jejuni responds to INT 407 and Caco-2 cells in a similar fashion at the cellular and molecular levels. Active protein synthesis was found to be required for C. jejuni to maximally invade these host cells. Proteomic and transcriptomic approaches were then used to define the protein and gene expression profiles of C. jejuni co-cultured with cells. By focusing on those genes showing increased expression by C. jejuni when co-cultured with epithelial cells, we discovered that C. jejuni quickly adapts to co-culture with epithelial cells by synthesizing gene products that enable it to acquire specific amino acids for growth, scavenge for inorganic molecules including iron, resist reactive oxygen/nitrogen species, and promote host cell interactions. Based on these findings, we selected a subset of the genes involved in chemotaxis and the regulation of flagellar assembly and generated C. jejuni deletion mutants for phenotypic analysis. Binding and internalization assays revealed significant differences in the interaction of C. jejuni chemotaxis and flagellar regulatory mutants. The identification of genes involved in C. jejuni adaptation to culture with host cells provides new insights into the infection process.

Safety profile of a replication-deficient human adenovirus-vectored foot-and-mouth disease virus serotype A24 subunit vaccine in cattle.

The safety of a replication-deficient, human adenovirus-vectored foot-and-mouth disease virus (FMDV) serotype A24 Cruzeiro capsid-based subunit vaccine (AdtA24) was evaluated in five independent safety studies. The target animal safety studies were designed in compliance with United States (U.S.) regulatory requirements (Title 9, U.S. Code of Federal Regulation [9CFR]) and international standard guidelines (VICH Topic GL-44) for veterinary live vaccines. The first three studies were conducted in a total of 22 vaccinees and demonstrated that the AdtA24 master seed virus (MSV) was safe, did not revert to virulence and was not shed or spread from vaccinees to susceptible cattle or pigs. The fourth safety study conducted in 10 lactating cows using an AdtA24 vaccine serial showed that the vaccine was completely absent from milk. The fifth safety study was conducted under typical U.S. production field conditions in 500 healthy beef and dairy cattle using two AdtA24 vaccine serials. These results demonstrated that the vaccine was safe when used per the product label recommendations. Additional data collected during these five studies confirmed that AdtA24 vaccinees developed FMDV A24 and the HAd5 vaccine vector serum neutralization antibodies that test negative in a FMDV non-structural protein antibody test, confirming AdtA24 vaccine's capability to differentiate infected from vaccinated animals (DIVA). In conclusion, results from this comprehensive set of cattle studies demonstrated the safety of the replication-deficient AdtA24 vaccine and fulfilled safety-related requirements for U.S. regulatory requirements.

Monitoring the evolution and migration of a methane gas plume in an unconfined sandy aquifer using time-lapse GPR and ERT.

Fugitive methane (CH4) leakage associated with conventional and unconventional petroleum development (e.g., shale gas) may pose significant risks to shallow groundwater. While the potential threat of stray (CH4) gas in aquifers has been acknowledged, few studies have examined the nature of its migration and fate in a shallow groundwater flow system. This study examines the geophysical responses observed from surface during a 72day field-scale simulated CH4 leak in an unconfined sandy aquifer at Canadian Forces Base Borden, Canada, to better understand the transient behaviour of fugitive CH4 gas in the subsurface. Time-lapse ground-penetrating radar (GPR) and electrical resistivity tomography (ERT) were used to monitor the distribution and migration of the gas-phase and assess any impacts to groundwater hydrochemistry. Geophysical measurements captured the transient formation of a CH4 gas plume emanating from the injector, which was accompanied by an increase in total dissolved gas pressure (PTDG). Subsequent reductions in PTDG were accompanied by reduced bulk resistivity around the injector along with an increase in the GPR reflectivity along horizontal bedding reflectors farther downgradient. Repeat temporal GPR reflection profiling identified three events with major peaks in reflectivity, interpreted to represent episodic lateral CH4 gas release events into the aquifer. Here, a gradual increase in PTDG near the injector caused a sudden lateral breakthrough of gas in the direction of groundwater flow, causing free-phase CH4 to migrate much farther than anticipated based on groundwater advection. CH4 accumulated along subtle permeability boundaries demarcated by grain-scale bedding within the aquifer characteristic of numerous Borden-aquifer multi-phase flow experiments. Diminishing reflectivity over a period of days to weeks suggests buoyancy-driven migration to the vadose zone and/or CH4 dissolution into groundwater. Lateral and vertical CH4 migration was primarily governed by subtle, yet measurable heterogeneity and anisotropy in the aquifer.

Dehydrogenative Transformations of Imines Using a Heterogeneous Photocatalyst.

Heterogeneous semiconductors are underexploited as photoredox catalysts in organic synthesis relative to their homogeneous, molecular counterparts. Here, we report the use of metal/TiO2 particles as catalysts for light-induced dehydrogenative imine transformations. The highly oxophilic nature of the TiO2 surface promotes the selective binding and dehydrogenation of alcohols in the presence of other oxidizable and Lewis basic functional groups. This feature enables the clean photogeneration of aldehyde equivalents that can be utilized in multicomponent couplings.

Multidimensional Investigation of Bedrock Heterogeneity/Unconformities at a DNAPL-Impacted Site.

Organic solvent (i.e., dense nonaqueous phase liquid, DNAPL) migration in the subsurface is known to be extremely sensitive to geologic heterogeneity. There is often a focus on heterogeneity that results from changing depositional conditions over short spatial scales. Similar or even more extreme spatial heterogeneity can result postdeposition due to erosional processes. This study applies a synergistic approach based on a combination of high-resolution lithologic logs of continuous cores, borehole geophysical logs, surface electrical resistivity, and seismic refraction tomography models to assess spatial heterogeneity in a shallow bedrock sequence subject to multiple unconformities and contaminated with a mixture of organic chemicals. The persistence of DNAPL in the source zone and an associated dissolved-phase plume led to variable impacts on formation resistivity across the study site. Seismic refraction in combination with electrical resistivity tomography improved interpretation of highly irregular erosional boundaries by delineating sharp lateral transitions in lithologic composition near the source zone and across the dissolved-phase plume. Electrical resistivity was effective at differentiating between clean and mud-rich sandstones and their unconformable contact with an underlying dolostone. Geophysical measurements revealed eroded dolostone mounds encased by a network of younger mud-rich sandstones channelized by clean semi-lithified sand, all of which was buried beneath variable glacial drift. Our synergistic multidimensional approach resulted in the development of a detailed three-dimensional shallow bedrock geospatial model, which has led to an improved understanding of DNAPL migration and contaminant plume heterogeneity.

Interruption of Electrical Conductivity of Titanium Dental Implants Suggests a Path Towards Elimination Of Corrosion.

Peri-implantitis is an inflammatory disease that results in the destruction of soft tissue and bone around the implant. Titanium implant corrosion has been attributed to the implant failure and cytotoxic effects to the alveolar bone. We have documented the extent of titanium release into surrounding plaque in patients with and without peri-implantitis. An in vitro model was designed to represent the actual environment of an implant in a patient's mouth. The model uses actual oral microbiota from a volunteer, allows monitoring electrochemical processes generated by biofilms growing on implants and permits control of biocorrosion electrical current. As determined by next generation DNA sequencing, microbial compositions in experiments with the in vitro model were comparable with the compositions found in patients with implants. It was determined that the electrical conductivity of titanium implants was the key factor responsible for the biocorrosion process. The interruption of the biocorrosion current resulted in a 4-5 fold reduction of corrosion. We propose a new design of dental implant that combines titanium in zero oxidation state for osseointegration and strength, interlaid with a nonconductive ceramic. In addition, we propose electrotherapy for manipulation of microbial biofilms and to induce bone healing in peri-implantitis patients.

The role of the Department of Homeland Security, Science and Technology Directorate in the development of vaccines and diagnostics for Transboundary Animal Diseases.

The development of countermeasures to support an effective response to Transboundary Animal Diseases (TAD) poses a challenge on a global scale and necessitates the coordinated involvement of scientists from government, industry and academia, as well as regulatory entities. The Agricultural Defense Branch under the Chemical and Biological Defense Division (CBD) of the Department of Homeland Security (DHS), Science and Technology Directorate (S&T) supports this important mission within the United States. This article provides an overview of the Agricultural Defense Branch's vaccine and diagnostic TAD project.

ON 01910.Na is selectively cytotoxic for chronic lymphocytic leukemia cells through a dual mechanism of action involving PI3K/AKT inhibition and induction of oxidative stress.

PURPOSE: Chronic lymphocytic leukemia (CLL), a malignancy of mature B cells, is incurable with chemotherapy. Signals from the microenvironment support leukemic cell survival and proliferation and may confer chemotherapy resistance. ON 01910.Na (Rigosertib), a multikinase phosphoinositide 3-kinase (PI3K) inhibitor, is entering phase III trials for myelodysplastic syndrome. Our aim was to analyze the efficacy of ON 01910.Na against CLL cells in vitro and investigate the molecular effects of this drug on tumor biology. EXPERIMENTAL DESIGN: Cytotoxicity of ON 01910.Na against CLL cells from 34 patients was determined in vitro with flow cytometry of cells stained with Annexin V and CD19. Global gene expression profiling on Affymetrix microarrays, flow cytometry, Western blotting, and cocultures with stroma cells were used to delineate ON 01910.Na mechanism of action. RESULTS: ON 01910.Na induced apoptosis in CLL B cells without significant toxicity against T cells or normal B cells. ON 01910.Na was equally active against leukemic cells associated with a more aggressive disease course [immunoglobulin heavy-chain variable region unmutated, adverse cytogenetics] than against cells without these features. Gene expression profiling revealed two main mechanisms of action: PI3K/AKT inhibition and induction of ROS that resulted in an oxidative stress response through activating protein 1 (AP-1), c-jun-NH(2)-terminal kinase, and ATF3 culminating in the upregulation of NOXA. ROS scavengers and shRNA mediated knockdown of ATF3- and NOXA-protected cells from drug-induced apoptosis. ON 01910.Na also abrogated the prosurvival effect of follicular dendritic cells on CLL cells and reduced SDF-1-induced migration of leukemic cells. CONCLUSIONS: These data support the clinical development of ON 01910.Na in CLL.

Laboratory evaluation of large-scale decontamination approaches.

AIMS: To evaluate the effectiveness of two spray-based decontamination methods for surface contamination reduction and to determine the potential for contamination spread by these methods. METHODS AND RESULTS: Material coupons (treated plywood and concrete) were contaminated with c. 1 × 10(7) spores of Bacillus atrophaeus by aerosol deposition. Decontaminants (pH-adjusted bleach or Spor-Klenz(®) RTU) were applied to coupons by either backpack sprayer or gas-powered sprayer. Contact time, reapplication frequency and rinse method were also varied. In addition to surface removal efficacy, partitioning of contamination between the rinsate and aerosol fractions was determined. Results indicated that pH-adjusted bleach was effective (≥6 logs reduction) when two applications and a 30 min contact time were administered, regardless of the decontaminant application method or material. Spor-Klenz(®) RTU was effective on wood, but achieved ≤3 logs reduction on concrete. A shortened application procedure with pH-adjusted bleach resulted in lower efficacy on wood, and a greater apparent potential for contamination spread. CONCLUSIONS: Consideration of material surface type is important when selecting a decontaminant. Also, achieving conditions that effectively inactivate surface biological contamination are critical to preventing the spread of contamination. SIGNIFICANCE AND IMPACT OF THE STUDY: Results presented here are intended to help development of remediation plans following a biological contamination incident.

Bortezomib resistance in mantle cell lymphoma is associated with plasmacytic differentiation.

Bortezomib induces remissions in 30%-50% of patients with relapsed mantle cell lymphoma (MCL). Conversely, more than half of patients' tumors are intrinsically resistant to bortezomib. The molecular mechanism of resistance has not been defined. We generated a model of bortezomib-adapted subclones of the MCL cell lines JEKO and HBL2 that were 40- to 80-fold less sensitive to bortezomib than the parental cells. Acquisition of bortezomib resistance was gradual and reversible. Bortezomib-adapted subclones showed increased proteasome activity and tolerated lower proteasome capacity than the parental lines. Using gene expression profiling, we discovered that bortezomib resistance was associated with plasmacytic differentiation, including up-regulation of IRF4 and CD38 and expression of CD138. In contrast to plasma cells, plasmacytic MCL cells did not increase immunoglobulin secretion. Intrinsically bortezomib-resistant MCL cell lines and primary tumor cells from MCL patients with inferior clinical response to bortezomib also expressed plasmacytic features. Knockdown of IRF4 was toxic for the subset of MCL cells with plasmacytic differentiation, but only slightly sensitized cells to bortezomib. We conclude that plasmacytic differentiation in the absence of an increased secretory load can enable cells to withstand the stress of proteasome inhibition. Expression of CD38 and IRF4 could serve as markers of bortezomib resistance in MCL. This study has been registered at http://clinicaltrials.gov as NCT00131976.

Molecularly defined antibody conjugation through a selenocysteine interface.

Antibody conjugates have broad utility in basic, preclinical, and clinical applications. Conventional antibody conjugation through the amine group of lysine or the thiol group of cysteine residues yields heterogeneous products of undefined stoichiometry and considerable batch-to-batch variability. To preserve the two hallmarks of the antibody molecule, precision and predictability, methods that enable site-specific antibody conjugation are in high demand. On the basis of a mammalian cell expression system, we describe the utilization of the 21st natural amino acid selenocysteine for the generation of IgG and Fab molecules with unique nucleophilic reactivity that affords site-specific conjugation to electrophilic derivatives of biotin, fluorescein, and poly(ethylene glycol). The resulting antibody conjugates were found to fully retain their antigen binding capability and, in the case of IgG, the ability to mediate effector functions. Gain of function was demonstrated in vitro and in vivo. While these antibody conjugates are relevant for a variety of proteomic, diagnostic, and therapeutic applications, they also constitute a proof of principle for the generation of molecularly defined antibody-drug conjugates and radioimmunoconjugates. Compared to other site-specific antibody conjugation methods, selenocysteine interface technology (i) only involves a minor modification at the C-terminus that does not interfere with disulfide bridges, (ii) does not require activation, and (iii) generates unique 1:1 stoichiometries of biological and chemical components. Collectively, our method affords the generation of highly defined antibody conjugates with broad utility from proteomic applications to therapeutic intervention.

Telemedicine is poised to revolutionize the practice of medicine.

Telemedicine--the application of audiovisual technology to patient care and medical education--has tremendous potential benefits, especially in linking doctors in remote rural locations to specialists in urban areas. The technology permits remote examinations and diagnoses of patients and continuing education for rural doctors. And there's potential for long-term savings by eliminating some hospital stays and reducing transportation costs. Telemedicine projects are multiplying as they become more economically feasible, thanks to new technology.