Viral inoculum dose impacts memory T‐cell inflation

Memory T‐cell inflation develops during certain persistent viral infections and is characterized by the accumulation and maintenance of large numbers of effector‐memory T cells, albeit with varying degrees in size and phenotype among infected hosts. The underlying mechanisms that control memory T‐cell inflation are not yet fully understood. Here, we dissected CMV‐specific memory T‐cell formation and its connection to the initial infectious dose by varying the inoculum size. After low dose inoculum with mouse CMV, the accumulation of inflationary memory T cells was severely hampered and correlated with reduced reservoirs of latent virus in nonhematopoietic cells and diminished antigen‐driven T‐cell proliferation. Moreover, lowering of the initial viral dose turned the characteristic effector memory‐like inflationary T cells into more central memory‐like cells as evidenced by the cell‐surface phenotype of CD27^high, CD62L⁺, CD127⁺, and KLRG1^?, and by improved secondary expansion potential. These data show the impact of the viral inoculum on the degree of memory T‐cell inflation and provide a rationale for the observed variation of human CMV‐specific T‐cell responses in terms of magnitude and phenotype.     

Thermotropic liquid crystals from biomacromolecules

Complexation of biomacromolecules (e.g., nucleic acids, proteins, or viruses) with surfactants containing flexible alkyl tails, followed by dehydration, is shown to be a simple generic method for the production of thermotropic liquid crystals. The anhydrous smectic phases that result exhibit biomacromolecular sublayers intercalated between aliphatic hydrocarbon sublayers at or near room temperature. Both this and low transition temperatures to other phases enable the study and application of thermotropic liquid crystal phase behavior without thermal degradation of the biomolecular components.Liquid crystals (LCs) play an important role in biology because their essential characteristic, the combination of order and mobility, is a basic requirement for self-organization and structure formation in living systems (1–3). Thus, it is not surprising that the study of LCs emerged as a scientific discipline in part from biology and from the study of myelin figures, lipids, and cell membranes (4). These and the LC phases formed from many other biomolecules, including nucleic acids (5, 6), proteins (7, 8), and viruses (9, 10), are classified as lyotropic, the general term applied to LC structures formed in water and stabilized by the distinctly biological theme of amphiphilic partitioning of hydrophilic and hydrophobic molecular components into separate domains. However, the principal thrust and achievement of the study of LCs has been in the science and application of thermotropic materials, structures, and phases in which molecules that are only weakly amphiphilic exhibit LC ordering by virtue of their steric molecular shape, flexibility, and/or weak intermolecular interactions [e.g., van der Waals and dipolar forces (11)]. These characteristics enable thermotropic LCs (TLCs) to adopt a wide variety of exotic phases and to exhibit dramatic and useful responses to external forces, including, for example, the electro-optic effects that have led to LC displays and the portable computing revolution. This general distinction between lyotropic LCs and TLCs suggests there may be interesting possibilities in the development of biomolecular or bioinspired LC systems in which the importance of amphiphilicity is reduced and the LC phases obtained are more thermotropic in nature. Such biological TLC materials are very appealing for several reasons. Most biomacromolecules were extensively characterized in aqueous environments, but in TLC phases, their solvent-free properties and functions could be investigated in a state in which no or only traces of water are present. Water exhibits a high dielectric constant and has the ability to form hydrogen bonds, greatly influencing the structure and functions of biomacromolecules or compromising electronic properties such as charge transport (12–15). Indeed, anhydrous TLC systems containing glycolipids (16–19), ferritin (20), and polylysine have been reported (21–23). However, a general approach to fabricating TLCs based on nucleic acids, polypeptides, proteins, and protein assemblies of large molecular weights such as virus particles remains elusive.Here we propose that the combination of biomaterials with suitably chosen surfactants, followed by dehydration, can be effectively applied as a simple generic scheme for producing biomacromolecular-based TLCs. We demonstrate that biological TLCs can be made from a remarkable range of biomolecules and bio-inspired molecules, including nucleic acids, polypeptides, fusion proteins, and viruses. TLC materials typically combine rigid or semirigid anisometric units, which introduce orientational anisotropy, with flexible alkyl chains, which suppress crystallization (24). In the present experiments, negatively charged biomolecules and bio-inspired molecules act as rigid parts, and cationic surfactants make up the flexible units to produce TLC phases with remarkably low LC-isotropic clearing temperatures, which is another TLC signature. Electrostatic interactions couple these rigid and flexible components into hybrid assemblies, which then order into lamellar phases of alternating rigid and flexible layers (Fig. 1) stabilized by the tendency in TLCs for rigid and flexible to spatially segregate (25).Open in a separate windowFig. 1.Proposed structures of TLCs formed by the biological building blocks complexed with surfactants, showing sketches of various lamellar phases and the corresponding phase transition temperatures (°C). The lamellar bilayer structures are made of, alternately, a sublayer of the biomacromolecules and an interdigitated sublayer of the surfactants, where the negatively charged parts of the biomolecules (e.g., phosphate groups of ssDNA and ssRNA, glutamate residues of supercharged ELPs, and N-terminal glutamate and aspartate residues of pVIII protein in phages) electrostatically interact with the cationic head groups of the surfactants. For the ssDNA–DOAB and ssRNA–DOAB smectic TLCs, the oligonucleotides are randomly orientated in the DNA (RNA) sublayers. For the ELP–DDAB complexes, in addition to the bilayer smectic phase, a modulated smectic (Sm_mod) phase is observed at lower temperature. For the phage–DOAB–DDAB lamellar structures, rodlike virus particles are embedded in a sublayer between interdigitated surfactants with additional in-plane orientational order.     

Deviant Strategy on the Modified Six Elements Test in Patients with Schizophrenia

Impaired executive functioning is found in a considerable proportion of schizophrenia patients. Neuropsychological tests are originally designed to measure the behavior of neurological patients and may therefore miss psychiatry-related cognitive deficits. Qualitative information on tests for executive functioning is important in psychiatric populations. The Modified Six Elements Test (MSET) is a planning test that consists of 6 tasks, for which subjects have limited time and have to obey to switching rules. This study concerns a qualitatively different approach schizophrenia patients use on the MSET, and its relationship with cognitive measures. MSET scores and strategies of schizophrenia patients were compared to those of healthy controls, closed-head-injury patients, and peripheral injury patients. Also, schizophrenia patients and healthy controls were compared on verbal memory and vigilance. Schizophrenia patients finish fewer assignments on the MSET, receive a lower profile score compared to healthy controls, and use a different strategy on the test compared to the other groups. They also perform below healthy controls on the tests for verbal memory and vigilance. Use of the different strategy in schizophrenia patients was related to impaired cognitive functioning. An interesting strategy used by schizophrenia patients on the MSET appears to be indicative of impaired cognitive functioning. This strategy may be a compensatory strategy to spare cognitive resources. It could also be the result of a concrete interpretation of the test instructions.     

Mapping Different Intra-Hemispheric Parietal-Motor Networks Using Twin Coil TMS

BackgroundAccumulating evidence suggests anatomical and functional differences in connectivity between the anterior and posterior parts of the inferior-parietal lobule (IPL) and the frontal motor areas.Objective/HypothesisThis study investigates whether different intra-hemispheric parietal-motor interactions can be observed along the anterior–posterior axis of the IPL in the resting human brain.MethodsWe use a twin coil transcranial magnetic stimulation technique to test intra-hemispheric interactions between three points adjacent to the intra-parietal sulcus (anterior, central, posterior) and the ipsilateral primary motor cortex (M1) at rest in both hemispheres.ResultsWe found that stimulation of the anterior IPL resulted in an inhibition of the ipsilateral M1 in both hemispheres. Stimulation of the central and posterior IPL resulted in a facilitatory effect on ipsilateral M1 in the left but not for the right hemisphere. Additionally we show that there is considerable inter-subject variability concerning the optimal parietal facilitatory and inhibitory position.ConclusionsThe IPL has distinct inhibitory and facilitatory connections to the ipsilateral M1. Whereas inhibitory connections are observed in both hemispheres, facilitatory connections are asymmetric. These parietal-motor networks may represent the basis for the functional differences between these regions in reaching and grasping tasks and mirror the functional asymmetry observed in the motor system. From a practical point of view, we note that the inter-subject variability means that future TMS studies of the parietal area might consider a hot-spot localization similar to the procedures commonly used for M1.     

Successful Treatment with Atomoxetine of an Adolescent Boy with Attention Deficit/Hyperactivity Disorder,Extreme Obesity,and Reduced Melanocortin 4 Receptor Function

ObjectiveRecent case reports suggest a link between reduced melanocortinergic tone and both obesity and attention deficit / hyperactivity disorder (ADHD). We present the case of a 13-year-old, male, obese MC4R mutation carrier with ADHD.Case ReportThe boy carries a heterozygous mutation in the melanocortin 4 receptor gene (MC4R; Met281Val), that leads to a reduced receptor function. Dominant mutations of this type represent major gene effects for obesity. He participated in a lifestyle intervention program for obesity and received treatment with the selective norepinephrine re-uptake inhibitor atomoxetine for 31 months. The boy markedly reduced his BMI from 47.2 to 29.6 kg/m².ConclusionAtomoxetine proved to efficiently reduce weight in a severely obese MC4R mutation carrier with ADHD. We briefly discuss possible mechanisms for our observation, including evidence for the functional connectivity between melanocortinergic, dopaminergic, and norepinephrinergic brain circuitries.Key Words: Pharmacogenetics, Brain-derived neurotrophic factor, Weight loss, Appetite control, Methylphenidate     

Dental composite materials containing carolacton inhibit biofilm growth of Streptococcus mutans

Objective

Caries adjacent to restorations is one of the main causes for restoration replacement. Antimicrobial substances incorporated into dental materials would potentially be able to reduce secondary caries initiation and progression. This study investigated biofilm growth of Streptococcus mutans UA159 on the surface of composite materials containing the biomolecule carolacton compared to materials containing chlorhexidine (CHX) and triclosan.

Methods

Biofilm inhibition was investigated by counting colony forming units (CFU), viability staining (Life/Dead), and real-time quantitative PCR.

Results

First, the antimicrobial substances were added to the cultivation medium at 2.5 μg/ml (0.0002%) and 0.25 μg/ml (0.00002%). CHX eliminated bacterial growth and biofilm formation completely. Triclosan was effective at 2.5 μg/ml, but at 0.25 μg/ml biofilm mass and viability were unchanged, yet the number of CFU increased due to disruption of cell chains and biofilm aggregates. Carolacton had a limited effect on biofilm growth and mass, but reduced viability significantly. When incorporated into composite materials carolacton (25 μg/ml resp. 0.002%, w/w) had no adverse effect on physical/mechanical properties and retained its biofilm inhibiting effect. Life/Dead staining revealed a reduction of biofilm viability of up to 64%. CFUs were reduced by 98% and qPCR demonstrated a mean inhibition of 87%. In contrast, materials containing CHX or triclosan showed an insignificant effect on biofilm formation, even at a 100fold increased concentration (0.2%). The anti-biofilm activity of composite material containing carolacton was stable over a period of 42 days.

Significance

Carolacton incorporated into dental filling material has a strong biofilm-inhibiting effect on S. mutans and is therefore potentially able to prevent secondary caries formation.     

Indications for and results of arthroscopy in the arthritic knee: a European survey

Purpose

The place of arthroscopic treatment in osteoarthritis of the knee has generated much controversy. A survey was initiated to collect the opinion of experienced surgeons.

Methods

Of the 211 surgeons interviewed, 170 (80.6 %) replied to the electronic questionnaire. Respondents had at least ten years of experience in arthroscopy and currently perform more than 100 arthroscopies per year. Various indications and treatment modalities for arthroscopy in osteoarthritis of the knee had to be evaluated on a scale from “excellent” to “no indication”.

Results

The respondents generally believe that an improvement is more likely in low-grade osteoarthritis (p < 0.001) and in neutral leg axis (p < 0.001). The outcome was rated better if symptoms had persisted for less than six months (p < 0.001) and for patients that were younger than 60 years (p < 0.001). Partial meniscectomy and notchplasty in cases of extension deficit were considered as successful treatment options. Debridement was an accepted indication, with an outcome mainly rated as fair. A majority saw no indication for joint lavage, arthroscopic treatment of arthrofibrosis and removal of osteophytes. The outcome appears to be poor if a bone edema is diagnosed on magnetic resonance imaging prior to arthroscopy. Only 55.9 % of respondents were comfortable with the current definition of osteoarthritis.

Conclusions

Experienced arthroscopic surgeons all over Europe believe arthroscopy in osteoarthritis is appropriate, under certain conditions. The major task for surgeons is to select the right patients who are likely to benefit from this intervention.