Retroviral Antisense Transcripts and Genes: 33 Years after First Predicted, a Silent Retroviral Revolution?

Paradigm shifts throughout the history of microbiology have typically been ignored, or met with skepticism and resistance, by the scientific community. This has been especially true in the field of virology, where the discovery of a "contagium vivum fluidum", or infectious fluid remaining after excluding bacteria by filtration, was initially ignored because it did not coincide with the established view of microorganisms. Subsequent studies on such infectious agents, eventually termed "viruses", were met with skepticism. However, after an abundance of proof accumulated, viruses were eventually acknowledged as defined microbiological entities. Next, the proposed role of viruses in oncogenesis in animals was disputed, as was the unique mechanism of genome replication by reverse transcription of RNA by the retroviruses. This same pattern of skepticism holds true for the prediction of the existence of retroviral "antisense" transcripts and genes. From the time of their discovery, it was thought that retroviruses encoded proteins on only one strand of proviral DNA. However, in 1988, it was predicted that human immunodeficiency virus type 1 (HIV-1), and other retroviruses, express an antisense protein encoded on the DNA strand opposite that encoding the known viral proteins. Confirmation came quickly with the characterization of the antisense protein, HBZ, of the human T-cell leukemia virus type 1 (HTLV-1), and the finding that both the protein and its antisense mRNA transcript play key roles in viral replication and pathogenesis. However, acceptance of the existence, and potential importance, of a corresponding antisense transcript and protein (ASP) in HIV-1 infection and pathogenesis has lagged, despite gradually accumulating theoretical and experimental evidence. The most striking theoretical evidence is the finding that asp is highly conserved in group M viruses and correlates exclusively with subtypes, or clades, responsible for the AIDS pandemic. This review outlines the history of the major shifts in thought pertaining to the nature and characteristics of viruses, and in particular retroviruses, and details the development of the hypothesis that retroviral antisense transcripts and genes exist. We conclude that there is a need to accelerate studies on ASP, and its transcript(s), with the view that both may be important, and overlooked, targets in anti-HIV therapeutic and vaccine strategies.

Multidrug-Resistant Hospital Bacteria: Epidemiological Factors and Susceptibility Profile.

Increase in antimicrobial resistance to antibiotics is the product of the evolution and natural adaptation of microorganisms through mutations and genetic recombination caused by the indiscriminate use of antibiotics and the ineffective control and prevention of infection. The current study analyzes the profile of multiresistant hospital bacteria in two hospitals in Pelotas, state of Rio Grande do Sul, Brazil. Over the course of 4 months, patient's gender and age, hospital accommodation type, and sample site were evaluated. Two hundred and eighty-six microbiological culture antibiogram reports of hospitalized patients and outpatients of both sexes, between zero and 96 years of age, were analyzed. Bacterium Klebsiella pneumoniae was the most prevalent. The most resistant Gram-negative bacilli (GNB) were K. pneumoniae (27.5%); Acinetobacter baumannii (24.1%); Escherichia coli (14.7%); and Pseudomonas aeruginosa (14.5%). The most resistant Gram-positive cocci (GPC) were Enterococcus faecium (27.5%) and Staphylococcus aureus (25.5%). The classes of antibiotics with the greatest number of resistant GNB included penicillins (84.8%), quinolones (77.5%), and cephalosporins (75.7%). In the case of GPC, the most resistant were macrolides (95.4%); lincosamides (90.3%), and penicillins (77%). Among GNBs, polypeptides had the highest sensitivity rate (81.3%), whereas, among GPC, fusidanes, glycylcyclines, and lipopeptides had 100% sensitivity.

Emergency Department and Inpatient Health Care Services Utilization by the Elderly Population: Hurricane Sandy in The State of New Jersey.

OBJECTIVE: In this investigation, we reported the increase in emergency department and inpatient admission cases during the month of November 2012 post Hurricane Sandy as compared with baseline (November 2010, 2011, and 2013) for elderly patients aged 65 and up. METHODS: Medical claims data for patients aged 65 and over treated at emergency department and inpatient health care facilities in New Jersey were analyzed to examine the surge in frequencies of diagnoses treated immediately following Hurricane Sandy. The differences were quantified using gap analysis for 2 years before and 1 year after the event. RESULTS: There was an average increase of 1700 cases for the month of November 2012 relative to baseline for the top 15 most frequently diagnosed emergency department medical conditions. On a daily basis, a volume increase by an average 57 cases could be expected, including significant numbers of limb fractures and other trauma cases for these most frequently encountered medical conditions. CONCLUSIONS: Understanding the surge level in medical services needed in emergency departments and inpatient facilities during a natural disaster aftermath is critical for effective emergency preparation and response for the elderly population. (Disaster Med Public Health Preparedness. 2018;12:730-738).

Understanding Patterns of Socioeconomic and Demographic Factors Along With Health Services Provider Availability for Zika Outbreak in South Florida.

In this study, we analyzed the patterns of socioeconomic and demographic factors along with health services provider availability for the current Zika outbreak in Miami-Dade County, South Florida. We used Center for Consumer Information & Insurance Oversight (CCIIO) Machine-Readable Public Use Files (MR-PUFs) to examine provider availability in combination with socioeconomic and demographic factors that could potentially lead to healthcare disparities between any underserved population of the Wynwood neighborhood and the broader population of Miami-Dade County. MR-PUFs contain public provider-level data from states that are participating in the Federally Facilitated Marketplace. According to CCIIO, an issuer of a Qualified Health Plan that uses a provider network must maintain a network that is sufficient in the number and types of providers, including providers that specialize in mental-health and substance-use disorder services, to assure that all services will be accessible to enrollees without unreasonable delay. (Disaster Med Public Health Preparedness. 2018;12:455-459).

Formation of Exotic Networks of Water Clusters in Helium Droplets Facilitated by the Presence of Neon Atoms.

Water clusters are formed in helium droplets via the sequential capture of monomers. One or two neon atoms are added to each droplet prior to the addition of water. The infrared spectrum of the droplet ensemble reveals several signatures of polar, water tetramer clusters having dipole moments between 2D and 3D. Comparison with ab initio computations supports the assignment of the cluster networks to noncyclic "3 + 1" clusters, which are ∼5.3 kcal/mol less stable than the global minimum nonpolar cyclic tetramer. The (H2O)3Ne + H2O ring insertion barrier is sufficiently large, such that evaporative helium cooling is capable of kinetically quenching the nonequilibrium tetramer system prior to its rearrangement to the lower energy cyclic species. To this end, the reported process results in the formation of exotic water cluster networks that are either higher in energy than the most stable gas-phase analogs or not even stable in the gas phase.

Reorganizing Nigeria's Vaccine Supply Chain Reduces Need For Additional Storage Facilities, But More Storage Is Required.

One of the major problems facing Nigeria's vaccine supply chain is the lack of adequate vaccine storage facilities. Despite the introduction of solar-powered refrigerators and the use of new tools to monitor supply levels, this problem persists. Using data on vaccine supply for 2011-14 from Nigeria's National Primary Health Care Development Agency, we created a simulation model to explore the effects of variance in supply and demand on storage capacity requirements. We focused on the segment of the supply chain that moves vaccines inside Nigeria. Our findings suggest that 55 percent more vaccine storage capacity is needed than is currently available. We found that reorganizing the supply chain as proposed by the National Primary Health Care Development Agency could reduce that need to 30 percent more storage. Storage requirements varied by region of the country and vaccine type. The Nigerian government may want to consider the differences in storage requirements by region and vaccine type in its proposed reorganization efforts.

The effects of repetitive vibration on sensorineural function: biomarkers of sensorineural injury in an animal model of metabolic syndrome.

Exposure to hand-transmitted vibration in the work-place can result in the loss of sensation and pain in workers. These effects may be exacerbated by pre-existing conditions such as diabetes or the presence of primary Raynaud's phenomena. The goal of these studies was to use an established model of vibration-induced injury in Zucker rats. Lean Zucker rats have a normal metabolic profile, while obese Zucker rats display symptoms of metabolic disorder or Type II diabetes. This study examined the effects of vibration in obese and lean rats. Zucker rats were exposed to 4h of vibration for 10 consecutive days at a frequency of 125 Hz and acceleration of 49 m/s(2) for 10 consecutive days. Sensory function was checked using transcutaneous electrical stimulation on days 1, 5 and 9 of the exposure. Once the study was complete the ventral tail nerves, dorsal root ganglia and spinal cord were dissected, and levels of various transcripts involved in sensorineural dysfunction were measured. Sensorineural dysfunction was assessed using transcutaneous electrical stimulation. Obese Zucker rats displayed very few changes in sensorineural function. However they did display significant changes in transcript levels for factors involved in synapse formation, peripheral nerve remodeling, and inflammation. The changes in transcript levels suggested that obese Zucker rats had some level of sensory nerve injury prior to exposure, and that exposure to vibration activated pathways involved in injury and re-innervation.

Strategic contracting practices to improve procurement of health commodities.

Public-sector entities responsible for procurement of essential medicines and health commodities in developing countries often lack the technical capacity to efficiently ensure supply security. Under strict public scrutiny and pressures to be transparent, many agencies continue to use archaic procurement methods and to depend on inflexible forecasts and cumbersome tendering processes. On the basis of semi-structured literature reviews and interviews, we identified framework agreements as a strategic procurement practice used by the U.S. federal government that may also be suitable for global health supply chains. Framework agreements are long-term contracts that provide the terms and conditions under which smaller repeat purchasing orders may be issued for a defined period of time. Such agreements are common in U.S. and United Nations procurement systems and in other developed countries and multilateral organizations. In contrast, framework agreements appear to be seldom used in procurement of health commodities in countries of sub-Saharan Africa. The current practice of floating tenders multiple times a year contributes to long lead times and stock-outs, and it hampers the manufacturer's or supplier's ability to plan and respond to the government's needs. To date, government's use of strategic contracting practices in public procurement of health commodities has not received much attention in most developing countries. It may present an opportunity for substantial improvements in procurement efficiency and commodity availability. Enabling legislation and strengthened technical capacity to develop and manage long-term contracts could facilitate the use of framework contracts in sub-Saharan Africa, with improved supply security and cost savings likely to result.

Update on immunizations in adults.

Vaccine-preventable diseases contribute significantly to the morbidity and mortality of U.S. adults. The Advisory Committee on Immunization Practices of the Centers for Disease Control and Prevention updates its recommended adult immunization schedule annually. The most recent updates include the permissive but not routine use of the quadrivalent human papillomavirus vaccine to prevent genital warts in males; a single dose of herpes zoster vaccine for adults 60 years and older, regardless of their history; replacing a single dose of tetanus and diphtheria toxoids (Td) vaccine with tetanus, diphtheria, and acellular pertussis (Tdap) vaccine in adults 19 years and older who have not previously received Tdap; expanding the indications for pneumococcal polyvalent-23 vaccine to include all adults with asthma and all smokers; annual seasonal influenza vaccination for all adults; and booster doses of meningococcal vaccine for adults with high-risk conditions. It is vital for family physicians to implement a systematic approach to adult immunization that is patient-, staff-, and physician-focused.

Vaccine supply chains need to be better funded and strengthened, or lives will be at risk.

In the next decade, at least twelve additional vaccines that target such diseases as typhoid, malaria, and dengue will become available to lower- and middle-income countries. These vaccines must travel along what are called supply chains, which include all personnel, systems, equipment, and activities involved in ensuring that vaccines are effectively delivered from the point of production to the people who need them. But for various reasons, supply chains are already strained in many developing countries, and the potential inability to distribute new vaccines will place lives at risk. Among the many steps needed to strengthen the global vaccine supply chain, we suggest that the international community pursue improved coordination between organizations that donate and ship vaccines and the host-country officials who receive and distribute the vaccines, as well as better training for supply-chain managers.

Formation of cold ion-neutral clusters using superfluid helium nanodroplets.

A strategy for forming and detecting cold ion-neutral clusters using superfluid helium nanodroplets is described. Sodium cations generated via thermionic emission are directed toward a beam of helium droplets that can also pick up neutral molecules and form a cluster with the captured Na(+). The composition of the clusters is determined by mass spectrometric analysis following a desolvation step. It is shown that the polar molecules H(2)O and HCN are picked up and form ion-neutral clusters with sizes and relative abundances that are in good agreement with those predicted by the statistics used to describe neutral cluster formation in helium droplets. [Na(H(2)O)(n)](+) clusters containing six to 43 water molecules were observed, a size range of sodiated water clusters difficult to access in the gas phase. Clusters containing N(2) were in lower abundance than expected, suggesting that the desolvation process heats the clusters sufficiently to dissociate those containing nonpolar molecules.

Challenge pools of hepatitis C virus genotypes 1-6 prototype strains: replication fitness and pathogenicity in chimpanzees and human liver-chimeric mouse models.

Chimpanzees represent the only animal model for studies of the natural history of hepatitis C virus (HCV). To generate virus stocks of important HCV variants, we infected chimpanzees with HCV strains of genotypes 1-6 and determined the infectivity titer of acute-phase plasma pools in additional animals. The courses of first- and second-passage infections were similar, with early appearance of viremia, HCV RNA titers of >10(4.7) IU/mL, and development of acute hepatitis; the chronicity rate was 56%. The challenge pools had titers of 10(3)-10(5) chimpanzee infectious doses/mL. Human liver-chimeric mice developed high-titer infections after inoculation with the challenge viruses of genotypes 1-6. Inoculation studies with different doses of the genotype 1b pool suggested that a relatively high virus dose is required to consistently infect chimeric mice. The challenge pools represent a unique resource for studies of HCV molecular virology and for studies of pathogenesis, protective immunity, and vaccine efficacy in vivo.

(HCN)(m)-M(n) (M = K, Ca, Sr): vibrational excitation induced solvation and desolvation of dopants in and on helium nanodroplets.

Infrared (IR) laser spectroscopy is used to probe the rotational and vibrational dynamics of the (HCN)(m)-M(n) (M = K, Ca, Sr) complexes, either solvated within or bound to the surface of helium nanodroplets. The IR spectra of the (HCN)(m)-K (m = 1-3), HCN-Sr, and HCN-Ca complexes have the signature of a surface species, similar to the previously reported spectra of HCN-M (M = Na, K, Rb, Cs) [Douberly, G. E.; Miller, R. E. J. Phys. Chem. A 2007, 111, 7292.]. A second band in the HCN-Ca spectrum is assigned to a solvated complex. The relative intensities of the two HCN-Ca bands are droplet size dependent, with the solvated species being favored in larger droplets. IR-IR double resonance spectroscopy is used to probe the interconversion of the two distinct HCN-Ca populations. While only a surface-bound HCN-Sr species is initially produced, CH stretch vibrational excitation results in a population transfer to a solvated state. Complexes containing multiple HCN molecules and one Sr atom are surface-bound, while the nu(1) (HCN)(2)Ca spectrum has both the solvated and surface-bound signatures. All HCN-(Ca,Sr)(n) (n > or = 2) complexes are solvated following cluster formation in the droplet. Density-functional calculations of helium nanodroplets interacting with the HCN-M show surface binding for M = Na with a binding energy of 95 cm(-1). The calculations predict a fully solvated complex for M = Ca. For M = Sr, a 2.2 cm(-1) barrier is predicted between nearly isoenergetic surface binding and solvated states.

Vibration disrupts vascular function in a model of metabolic syndrome.

Vibration-induced white finger (VWF) is a disorder seen in workers exposed to hand-transmitted vibration, and is characterized by cold-induced vasospasms and finger blanching. Because overweight people with metabolic syndrome are pre-disposed to developing peripheral vascular disorders, it has been suggested that they also may be at greater risk of developing VWF if exposed to occupational vibration. We used an animal model of metabolic syndrome, the obese Zucker rat, to determine if metabolic syndrome alters vascular responses to vibration. Tails of lean and obese Zucker rats were exposed to vibration (125 Hz, 49 m/s(2) r.m.s.) or control conditions for 4 h/d for 10 d. Ventral tail arteries were collected and assessed for changes in gene expression, levels of reactive oxygen species (ROS) and for responsiveness to vasomodulating factors. Vibration exposure generally reduced the sensitivity of arteries to acetylcholine (ACh)-induced vasodilation. This decrease in sensitivity was most apparent in obese rats. Vibration also induced reductions in vascular nitric oxide concentrations and increases in vascular concentrations of ROS in obese rats. These results indicate that vibration interferes with endothelial-mediated vasodilation, and that metabolic syndrome exacerbates these effects. These findings are consistent with idea that workers with metabolic syndrome have an increased risk of developing VWF.

Study of the CH3 H2O radical complex stabilized in helium nanodroplets.

The weakly bound CH(3)H(2)O radical complex has been investigated by infrared laser spectroscopy. The complex is stabilized in helium nanodroplets and prepared by sequential pick up of a methyl radical and water molecule. Partially rotationally resolved spectra corresponding to the v = 1 <-- 0 excitation of the symmetric H(2)O stretching vibration within the complex show a significant red shift (25.06 cm(-1)) when compared with the symmetric stretch of H(2)O monomer, in agreement with the hydrogen bonded like structure derived by theory. Additional broad features were observed in the region predicted by theory for the antisymmetric stretch supporting our assignment. The B rotational constant is found to be 3.03 times smaller than predicted by ab initio calculations, with the reduction being attributed to the effects of helium solvation. The permanent electric dipole moment of the complex is experimentally determined to be 2.1 +/- 0.3 D using Stark spectroscopy. Ab initio calculations are also reported that provide support to the experimental results, as well as investigate the nature of large amplitude vibrational motion within the complex.

High-resolution infrared spectroscopy of Mg-HF and Mg-(HF)2 solvated in helium nanodroplets.

High-resolution infrared (IR) spectroscopy is used to investigate the Mg-HF and Mg-(HF)(2) van der Waals complexes. Both complexes are formed and probed within helium nanodroplets. Rotationally resolved zero-field and Stark spectra are assigned to a linear binary complex composed of a Mg atom bound to the hydrogen end of the HF molecule. Although high level ab initio calculations predict a fluorine bonded complex, none of the observed IR bands can be assigned to this complex. The collocation method is employed to determine the bound states on the two-dimensional intermolecular Mg-HF potential energy surface. The ground and first excited state wave functions for this potential surface have zero amplitude in the well corresponding to the fluorine bonded complex, consistent with experiment. The two HF stretching bands of the Mg-(HF)(2) complex are observed and assigned using a combination of the spectral symmetry, ab initio calculations, pick-up cell pressure dependencies, and dipole moment measurements. Comparisons with the helium solvated HF dimer show large changes to the HF stretching frequencies upon the addition of a single Mg atom to the hydrogen side of (HF)(2).

Increased oxidant activity mediates vascular dysfunction in vibration injury.

Occupational exposure to hand-operated vibrating tools causes a spectrum of pathological changes in the vascular, neurological, and musculoskeletal systems described as the hand-arm vibration syndrome (HAVS). Experiments were performed to determine the effects of acute vibration on the function of digital arteries. Rats paws were exposed to a vibrating platform (4 h, 125 Hz, constant acceleration of 49 m/s(2) root mean squared), and digital artery function was assessed subsequently in vitro using a pressure myograph system. Constriction to phenylephrine or 5-hydroxytryptamine was reduced in digital arteries from vibrated paws. However, after endothelium denudation, constriction to the agonists was no longer impaired in vibrated arteries. Inhibition of nitric-oxide synthase (NOS) with N(omega)-nitro-l-arginine methyl ester (l-NAME) increased constriction to phenylephrine or 5-hydroxytryptamine in vibrated but not control arteries and abolished the vibration-induced depression in constrictor responses. However, nitric oxide (NO) activity, determined using the NO-sensitive probe 4-amino-5-methylamino-2', 7'-difluorofluorescein, was reduced in vibrated compared with control arteries. Endogenous levels of reactive oxygen species (ROS), determined using the ROS-sensitive probe 5-(and 6)-chloromethyl-2',7'-dichlorodihydro-fluorescein, were increased in vibrated compared with control arteries. The increased ROS levels were abolished by L-NAME or by catalase, which degrades extracellular hydrogen peroxide. Catalase also increased constriction to phenylephrine or 5-hydroxytryptamine in vibrated but not control arteries and abolished the vibration-induced depression in constrictor responses. The results suggest that acute vibration causes vascular dysfunction in digital arteries by increasing ROS levels, which is probably mediated by uncoupling of endothelial NOS. Therefore, therapeutic strategies to inhibit ROS or augment NO activity may be beneficial in HAVS.

Ionization and fragmentation of isomeric van der Waals complexes embedded in helium nanodroplets.

The ionization and charge transfer processes, which occur when a doped helium droplet undergoes electron impact, are studied for droplets doped with van der Waals complexes with various structures and electrostatic moments. The mass spectra of the two isomers of hydrogen cyanide complexed with either cyanoacetylene or acetylene in helium droplets were obtained using optically selected mass spectrometry, and show that the structure of the complex has a large effect on the fragmentation pattern. The resulting fragmentation pattern is consistent with an ionization process in which charge steering strongly influences the site of initial ionization. The observed dissociation products may also be subject to caging by the helium matrix.

High prevalence of nasopharyngeal colonization by Staphylococcus aureus among children with HIV-1 infection in extreme southern Brazil.

OBJECTIVES: To compare nasopharyngeal colonization between children with HIV-1 infection and those without HIV-1 infection, with special emphasis on nasopharyngeal carriage of Staphylococcus aureus. METHODS: This hospital-based cross-sectional study was carried out in the Paediatric Day Hospital of a teaching hospital. Nasopharyngeal swabs were collected in 93 children aged up to 18 years old born to HIV-positive mothers (31 children with HIV-1 infection and 62 age-matched non-infected children). RESULTS: The prevalence of nasopharyngeal colonization by S. aureus was higher among children with HIV-1 infection compared with those without HIV-1 infection (45.16% vs. 12.9%, p = 0.001). After adjusting all potential confounders, HIV-1 infection was an independent risk factor for nasopharyngeal colonization by S. aureus, with a prevalence ratio of 4.29 (95% confidence interval: 1.72-10.70). CONCLUSION: Children with HIV-1 infection had a higher prevalence of nasopharyngeal colonization by S. aureus than children without HIV-1 infection. Most of the isolated strains of S. aureus were methicillin-susceptible.

Nonplanarity of adenine: vibrational transition moment angle studies in helium nanodroplets.

Mid-infrared spectra are reported for adenine monomer in helium nanodroplets. We show that there is only one tautomer of adenine, the global minimum structure, observed in helium nanodroplets and characterized by using ab initio calculations and the measurement of vibrational transition moment angles (VTMAs) for the various vibrational modes of the adenine monomer. On the basis of the VTMA analysis on the amino group of the global minimum tautomer, which gives insights into its nonplanarity, a detailed VTMA study of three lowest-energy amino tautomers of adenine is discussed in this study.