Quiescent T lymphocytes as an inducible virus reservoir in HIV-1 infection

To better understand the basis for human immunodeficiency virus type 1 (HIV-1) persistence and latency, the form in which viral DNA exists in the peripheral T lymphocyte reservoir of infected individuals was investigated. In asymptomatic individuals, HIV-1 was harbored predominantly as full-length, unintegrated complementary DNA. These extrachromosomal DNA forms retained the ability to integrate upon T cell activation in vitro. In patients with acquired immunodeficiency syndrome (AIDS), there was an increase in integrated relative to extrachromosomal DNA forms. By analysis of DNA from patient lymphocyte subpopulations depleted of human lymphocyte antigen-Dr receptor-positive cells, quiescent T cells were identified as the source of extrachromosomal HIV-1 DNA. Thus quiescent T lymphocytes may be a major and inducible HIV-1 reservoir in infected individuals.     

Endothelial cell membranes: polarity of particles as seen by freeze-fracturing

Freeze-fracturing shows particles within membranes. In plasma membranes of most cells the particles are more strongly bound to the inner half. In unfixed endothelial cells, this polarity is reversed. Glutaraldehyde fixation results in conventional polarity. The reverse polarity may be related to a mechanism for preferential fusion of pinocytotic vesicles with the plasma membrane.     

Differences in ascorbate and glutathione levels as indicators of resistance and susceptibility in Eucalyptus trees infected with Phytophthora cinnamomi

In this study we investigated the role that ascorbate (AA) and glutathione (GSH) play in the plant pathogen interaction of susceptible Eucalyptus sieberi L. A. Johnson and resistant Eucalyptus sideroxylon Woolls with Phytophthora cinnamomi Rands root infection. In a glasshouse study, seedlings were grown in soil-free plant boxes to facilitate the inoculation of the root systems by a P. cinnamomi zoospore solution. Ascorbate and GSH concentrations were measured in infected roots and leaves, along with leaf gas exchange, chlorophyll fluorescence and carbohydrate concentrations over a time course up to 312 h (13 days) post-inoculation (pi). At the early stages of infection (from 24 h pi), significant decreases in AA and GSH concentrations were observed in the infected roots and leaves of the susceptible E. sieberi seedlings. At the later stage of infection (312 h pi), the earlier AA decreases in the leaves of infected plants had become significant increases. In contrast, late, significant AA increases in the absence of any GSH changes were observed in the infected roots of the resistant E. sideroxylon seedlings. In E. sideroxylon leaves, a significant GSH increase occurred at 24 h pi; however, by 312 h pi the earlier increase had become a significant decrease, while no changes occurred in AA. In E. sieberi, photosynthesis (A), stomatal conductance (g(s)) and PSII quantum efficiency (Φ(PSII)) were reduced by ～60, 80 and 30%, respectively, in infected plants and remained significantly lower than uninfected controls for the duration of the experiment. Significant reductions in these parameters did not occur until later (120 h pi for g(s) and 312 h pi for A and Φ(PSII)), and to a lesser extent in the resistant species. Non-structural carbohydrate analysis of roots and leaves indicate that carbohydrate metabolism and resource flow between shoots and roots may have been altered at later infection stages. This study suggests that reduced antioxidant capacity, leaf physiological function and carbohydrate metabolism are associated with susceptibility in E. sieberi to P. cinnamomi infection, while AA increases and new root formation were associated with resistance in E. sideroxylon.     

Lactose Synthase Components in Milk: Concentrations of α-Lactalbumin and β1,4-Galactosyltransferase in Milk of Cows from Several Breeds at Various Stages of Lactation

It is believed that milk production is determined by the number and activity of mammary secretory cells. Secretory activity, as assessed by milk volume, depends on secretion of the major osmole in milk, lactose, which is produced by lactose synthase. The amount of either of the two proteins in lactose synthase may regulate milk production. The objective of this study was to determine whether the concentrations in milk of the two components of lactose synthase, α‐lactalbumin (α‐LA) and β1,4‐galactosyltransferase (B4GALT), were related to genetic background, stage of lactation, breed or parity of dairy cows. α‐Lactalbumin and B4GALT concentrations were measured by ELISA and by enzyme assays, respectively, from single milk samples. Two herds with a total of 279 cows were used in the analysis. One herd contained Ayrshire, Brown Swiss, Holstein and Jersey cows; the second herd contained two groups of cows; Holsteins selected for high milk production and Holsteins with 1960s genetics. The α‐LA concentration in milk was greater in Jerseys and Ayrshires than in Holsteins and Brown Swiss. However, no difference in α‐LA concentration was observed in milk from high and low genetic merit cows in the Minnesota herd or among different genetic backgrounds in the Illinois herd. β1,4‐Galactosyltransferase concentrations were similar for all groups that were analyzed. α‐Lactalbumin concentrations were positively correlated with milk protein concentration, milk fat concentration and lactose concentration. β1,4‐Galactosyltransferase concentration in milk exhibited a strong positive correlation with number of days in milk. Although the concentration of B4GALT increased as lactation progressed, the values did not show any correlation with persistency of lactation or late lactation milk production. In conclusion, this survey shows that the two components of lactose synthase are each correlated to protein concentration and individually correlated to the concentration of other milk components and stage of lactation.     

Organogenesis and foetal haemodynamics during the normal gestation of healthy black‐rumped agoutis (Dasyprocta prymnolopha,Wagler, 1831) bred in captivity

The objective of this study was to define the patterns of organogenesis and foetal haemodynamics during the normal gestation of healthy agoutis (Dasyprocta prymnolopha) kept in captivity. Thirty pregnant agoutis that ranged in size from small to medium and weighed between 2.5 and 3 kg underwent B‐mode and Doppler ultrasonography for the biometric evaluation of the foetal organs. The foetal aortic blood flow proved to be predominantly systolic, and the measured flow velocity was 78.89 ± 2.95 cm/s, with a maximum pressure gradient of 2.12 ± 0.27 mmHg. The liver was characterized by its large volume, occupying the entire cranial aspect of the abdominal cavity, and it was associated cranially with the diaphragm and caudally with the stomach. The flow velocity in the portal vein was estimated to equal 12.17 ± 2.37 cm/s, with a resistivity index of 0.82 ± 0.05. The gallbladder was centrally located and protruded cranially towards the diaphragm. The spleen was visualized as an elongated structure with tapered cranial and caudal extremities, and the foetal kidneys were visualized bilaterally in the retroperitoneal region, with the right kidney positioned slightly more cranially than the left. The morphological characterization and hemodynamic analysis of the foetal organs of black‐rumped agoutis via B‐mode and Doppler ultrasonography allow determination of the vascular network and of reference values for the blood flow required for perfusing the anatomical elements essential for maintaining the viability of foetuses at different gestational ages.     

Exogenous DNA Uptake of Boar Spermatozoa by a Magnetic Nanoparticle Vector System

The sperm‐mediated gene transfer method is applicable to transgenesis in many species that use spermatozoa for reproduction recently, which has been shown various results. In the current study, we show that transgenic porcine embryos can be efficiently produced by employing a simple transfection method that uses magnetic nanoparticles (MNPs). The complexes formed between plasmid DNA and MNPs were bounded on ejaculated boar spermatozoa at a higher efficiency compared to methods using DNA alone or lipofection. Using confocal microscopy, rhodamine fluorophore‐labelled MNPs were detected on external surfaces of the spermatozoa membrane, which were bounded on zona pellucida of in vitro maturated oocyte during in vitro fertilization. Electron microscopy revealed that clusters of MNPs were detected in inside of plasma membrane and nucleus of the spermatozoa head. Additionally, we found that magnetofected boar spermatozoa could be fertilized with oocytes in vitro and that the resulting gene of green fluorescent protein was detected in fertilized eggs by genomic PCR analysis. Taken together, these results suggest that MNPs can be used to efficiently introduce a transgene into embryo via spermatozoa.     

Earthworms increase the ratio of bacteria to fungi in northern hardwood forest soils, primarily by eliminating the organic horizon

The exotic earthworm invasion in hardwood forests of the northern United States is associated with many ecosystem-level changes. However, less is known about the effects of the invasion on the composition of the soil microbial community through which ecosystem-level changes are mediated. Further, earthworm effects on soil microbial community composition have not been well studied in the field. To evaluate changes in bacterial and fungal abundance associated with the earthworm invasion we quantified bacterial and fungal biomass by microscopic counts in paired earthworm-invaded (earthworm) and earthworm-free (reference) plots in five forest stands in central New York (USA). Earthworms significantly increased the ratio of bacteria to fungi on an area basis (per m²), by more than two times in mid-summer and early autumn. While this effect was associated primarily with the lack of the fungal-dominated organic horizon in earthworm plots, a higher ratio of bacteria to fungi in the surface 5 cm mineral soil also contributed as it developed between spring and mid-summer. Earthworm reduction of fungal biomass was confirmed by substantially lower growth of fungal hyphae into mesh sand bags in earthworm compared to reference plots. Burrowing activity by the earthworm Lumbricus terrestris increased the ratio of bacteria to fungi over the short-term within earthworm plots, introducing small-scale spatial heterogeneity associated with burrows. Our study suggests that the exotic earthworm invasion in these northern hardwood forests markedly increased the ratio of bacteria to fungi by eliminating the fungal-rich organic horizon, and was associated localized increases in bacterial vs. fungal abundance in mineral soil, setting the stage for future research into linkages between the earthworm invasion, bacterial and fungal abundance, and ecosystem processes.     

Minimally invasive approach to lumbosacral decompression in a cadaveric canine model

AIMS: To determine efficacy of a minimally invasive (MI) surgical approach using a human MI lumbar retractor for canine lumbosacral dorsal laminectomy and partial discectomy and to compare this technique to the standard open surgical (OS) approach.

METHODS: Lumbosacral dorsal laminectomy and partial discectomy was performed on 16 large-breed canine cadavers using either a standard OS (n=8) or MI (n=8) approach. Skin and fascial incision length, procedure time, and intraoperative complications were recorded. Postoperatively specimens were evaluated for laminectomy and discectomy dimensions, and visible damage to the cauda equina and exiting nerve roots.

RESULTS: Median length of skin and fascial incisions in the OS group were longer than in the MI group (p<0.001). Median laminectomy length was similar between both approaches (p=0.234) but width was greater for the MI than OS approach (p=0.002). Both approaches achieved similar partial discectomy width (p=0.279). Overall surgical time was longer for MI approaches compared to OS, with a median of 18.5 (min 15.5, max 21.8) minutes for MI compared to 14.6 (min 13.1, max 16.9) minutes for OS (p=0.001).

CONCLUSIONS: The MI approach reduced incision lengths while retaining comparable laminectomy and discectomy dimensions. For this in vitro model the MI approach required more time to complete, but this difference may not be relevant in clinical cases.

CLINICAL RELEVANCE: Dogs undergoing lumbosacral dorsal laminectomy are commonly large-breed dogs. The traditional open approach requires a large skin incision and soft tissue dissection, especially in overweight animals. A MI approach accomplishing the same surgical result while minimising soft tissue trauma could reduce post-operative pain and recovery time, and may lower wound-related complications. Clinical studies are needed to confirm postoperative benefit and assess operating times in vivo.     

Defining place-keeping: The long-term management of public spaces

There has long been a focus in urban landscape planning and design on the creation of highquality public spaces, or place-making. Large amounts of capital continue to be spent on creating such spaces without adequate thought or resources for their long-term maintenance and management of public spaces, or place-keeping.While there may be continued policy rhetoric about the importance of place-keeping, particularly as public spaces are recognised for their important contribution to health, wellbeing, biodiversity and also their economic value, this has not however been supported in practice. There are examples in many cities where public spaces are subject to poor management and maintenance practices. This is clearly visible where vandalism, litter and damage to facilities and equipment occur, and people no longer feel safe or comfortable.This paper argues that this uneven focus on place-keeping is not only due to a lack of resources but also a lack of understanding of the concept, its complexity and the wide implications it has for users, practitioners and policymakers. The paper aims to address this gap in knowledge by providing a detailed exploration and definition of place-keeping within the urban context. Place-keeping is not simply about the physical environment, its design and maintenance, but also encompasses the interrelated and non-physical dimensions of partnerships, governance, funding, policy and evaluation.