ESCRT recruitment to SARS-CoV-2 spike induces virus-like particles that improve mRNA vaccines.

Prime-boost regimens for COVID-19 vaccines elicit poor antibody responses against Omicron-based variants and employ frequent boosters to maintain antibody levels. We present a natural infection-mimicking technology that combines features of mRNA- and protein nanoparticle-based vaccines through encoding self-assembling enveloped virus-like particles (eVLPs). eVLP assembly is achieved by inserting an ESCRT- and ALIX-binding region (EABR) into the SARS-CoV-2 spike cytoplasmic tail, which recruits ESCRT proteins to induce eVLP budding from cells. Purified spike-EABR eVLPs presented densely arrayed spikes and elicited potent antibody responses in mice. Two immunizations with mRNA-LNP encoding spike-EABR elicited potent CD8+ T cell responses and superior neutralizing antibody responses against original and variant SARS-CoV-2 compared with conventional spike-encoding mRNA-LNP and purified spike-EABR eVLPs, improving neutralizing titers >10-fold against Omicron-based variants for 3 months post-boost. Thus, EABR technology enhances potency and breadth of vaccine-induced responses through antigen presentation on cell surfaces and eVLPs, enabling longer-lasting protection against SARS-CoV-2 and other viruses.

Genetic and phenotypic characterization of NKX6-2-related spastic ataxia and hypomyelination.

BACKGROUND AND PURPOSE: Hypomyelinating leukodystrophies are a heterogeneous group of genetic disorders with a wide spectrum of phenotypes and a high rate of genetically unsolved cases. Bi-allelic mutations in NKX6-2 were recently linked to spastic ataxia 8 with hypomyelinating leukodystrophy. METHODS: Using a combination of homozygosity mapping, exome sequencing, and detailed clinical and neuroimaging assessment a series of new NKX6-2 mutations in a multicentre setting is described. Then, all reported NKX6-2 mutations and those identified in this study were combined and an in-depth analysis of NKX6-2-related disease spectrum was provided. RESULTS: Eleven new cases from eight families of different ethnic backgrounds carrying compound heterozygous and homozygous pathogenic variants in NKX6-2 were identified, evidencing a high NKX6-2 mutation burden in the hypomyelinating leukodystrophy disease spectrum. Our data reveal a phenotype spectrum with neonatal onset, global psychomotor delay and worse prognosis at the severe end and a childhood onset with mainly motor phenotype at the milder end. The phenotypic and neuroimaging expression in NKX6-2 is described and it is shown that phenotypes with epilepsy in the absence of overt hypomyelination and diffuse hypomyelination without seizures can occur. CONCLUSIONS: NKX6-2 mutations should be considered in patients with autosomal recessive, very early onset of nystagmus, cerebellar ataxia with hypotonia that rapidly progresses to spasticity, particularly when associated with neuroimaging signs of hypomyelination. Therefore, it is recommended that NXK6-2 should be included in hypomyelinating leukodystrophy and spastic ataxia diagnostic panels.

Does meal-time insulin dosing based on fat-protein counting give positive results in postprandial glycaemic profile after a high protein-fat meal in adolescents with type 1 diabetes: a randomised controlled trial.

BACKGROUND: The present study investigated the effects of fat-protein (CFP) counting in addition to carbohydrate (CARB) counting for calculating prandial insulin dosage on blood glucose profile in patients with type 1 diabetes (T1D) on basal-bolus insulin therapy. METHODS: In this single centre, cross-over, randomised, controlled study, control meal (SM: standard meal using a carbohydrate counting method) and three test meals (HPM: high protein meal using a carbohydrate counting method; HPFM-a: high protein-fat meal using a carbohydrate counting method; HPFM-b: high protein-fat meal using a carbohydrate and fat-protein counting method) were compared on postprandial early (0-120 min), late (120-240 min) and total (0-240 min) glucose response in 30 patients with T1D, aged 16-18 years. RESULTS: The glucose levels of 0-90 min did not change after different meal consumptions (P > 0.05), whereas 120-240 min glucose levels were higher after HPFM-a consumption compared to HPFM-b consumption (P < 0.05). There were no significant differences between meals with respect to the early postprandial glucose response (0-120 min) (P = 0.405). In late response (120-240 min), HPFM-b [area under the curve (AUC) = 20 609 (582) mg dL-1  × dk] was significantly lower than SM [AUC = 24 092 (9015) mg dL-1  × dk], HPM [AUC = 24 072 (5853) mg dL-1  × dk] and HPFM-a [AUC = 25 986 (6979) mg dL-1  × dk] (P = 0.032). CONCLUSIONS: Meal-related insulin dosing based on carbohydrate plus fat/protein counting has given positive results in the postprandial glycaemic profile as a result of lower postprandial glycaemic levels compared to conventional carbohydrate counting in patients with T1D after a high protein-fat meal.

Association between attitudes towards Internet usage and health practices in high-school students in 2 schools in Turkey: a cross-sectional study.

Excessive usage of the Internet can negatively affect health and health practices especially among youth. To examine the issue among Turkish students, this study determined the association between high-school students' attitudes towards Internet usage and their health behaviour. The sample (n = 2043) was randomly drawn from all students at two high schools in Turkey: one in Anatolia and one (a vocational school) in Istanbul. Data were collected using the Structured Questionnaire, Attitude Scale towards Internet Usage (ASTIU) and Adolescent Lifestyle Profile (ALP). The mean age of the students was 16.3 years, 77.7% were male and 96.9% used the Internet; mean duration of Internet use was 18.8 h/week. Female students had more positive attitudes towards Internet usage. Mean scores for ASTIU and ALP were 72.58 (SD 17.64) and 111.34 (SD 16.61) respectively, which were average levels. There was a statistically significant but weak overall negative correlation between adolescents' attitudes towards Internet usage and their health behaviour. Our results concur with studies in different cultures which suggest some negative effects, of heavy Internet use.

Treatment of Geriatric Proximal Humerus Fractures: Indications and Outcome.

Treatment algorithms of proximal humerus fractures are still controversially discussed. The enthusiasm towards operative treatment after the introduction of locking implants, has not been justified by the functional results in the elderly population. The majority of those fractures in the geriatric patients can be successfully treated conservatively. Thorough clinical and radiological examination for fracture analysis, dynamic stability control with the use of an image intensifier, and meticulous reduction, in addition with the appropriate orthesis for its retention and rehabilitation, are the keys for the successful treatment of the proximal humerus fracture in the elderly patient. The present review reports on the main treatment aspects of proximal humerus fractures in the geriatric population and proposes a treatment algorithm.

Probing nonadiabatic molecular alignment by spectral modulation.

We investigated molecular alignment wakes of femtosecond laser pulses. Evolution of nonadiabatic molecular alignment in nitrogen gas has been measured via its nonlinear interaction effects with a variably delayed probe pulse. The induced rotational wave packet was mapped as a function of the angular difference between polarization directions of femtosecond pump and probe pulses as well as their relative delay and the plot of the variations of the rotational wave packet, i.e. "quantum carpet", was found to be in good agreement with the calculated angular and temporal dependencies of molecular alignment parameter.

White-light generation control with crossing beams of femtosecond laser pulses.

We investigated the variations in generated white-light when crossing two femtosecond laser beams in a Kerr medium. By changing the relative delay of two interacting intense femtosecond laser pulses, we show that white-light generation can be enhanced or suppressed. With a decrease of the relative delay an enhancement of the white-light output was observed, which at even smaller delays was reverted to a suppression of white-light generation. Under choosen conditions, the level of suppression resulted in a white-light output lower than the initial level corresponding to large delays, when the pulses do not overlap in time. The enhancement of the white-light generation takes place in the pulse that is lagging. We found that the effect of the interaction of the beams depends on their relative orientation of polarization and increases when the polarizations are changed from perpendicular to parallel. The observed effects are explained by noting that at intermediate delays, the perturbations introduced in the path of the lagging beam lead to a shortening of the length of filament formation and enhancement of the white-light generation, whereas at small delays the stronger interaction and mutual rescattering reduces the intensity in the central part of the beams, suppressing filamentation and white-light generation.

Proactive vaccination using multiviral Quartet Nanocages to elicit broad anti-coronavirus responses.

Defending against future pandemics requires vaccine platforms that protect across a range of related pathogens. Nanoscale patterning can be used to address this issue. Here, we produce quartets of linked receptor-binding domains (RBDs) from a panel of SARS-like betacoronaviruses, coupled to a computationally designed nanocage through SpyTag/SpyCatcher links. These Quartet Nanocages, possessing a branched morphology, induce a high level of neutralizing antibodies against several different coronaviruses, including against viruses not represented in the vaccine. Equivalent antibody responses are raised to RBDs close to the nanocage or at the tips of the nanoparticle's branches. In animals primed with SARS-CoV-2 Spike, boost immunizations with Quartet Nanocages increase the strength and breadth of an otherwise narrow immune response. A Quartet Nanocage including the Omicron XBB.1.5 'Kraken' RBD induced antibodies with binding to a broad range of sarbecoviruses, as well as neutralizing activity against this variant of concern. Quartet nanocages are a nanomedicine approach with potential to confer heterotypic protection against emergent zoonotic pathogens and facilitate proactive pandemic protection.

A novel syndrome of abnormal striatum and congenital cataract: evidence for linkage to chromosomes 11.

We report a consanguineous family of three girls and one boy affected with a novel syndrome involving the lens and the basal ganglia. The phenotype is strikingly similar between affected siblings with cognitive impairment, attention deficit hyperactivity disorder (ADHD), microcephaly, growth retardation, congenital cataract, and dystonia. The magnetic resonance imaging showed unusual pattern of swelling of the caudate heads and thinning of the putamina with severe degree of hypometabolism on the [18F] deoxyglucose positron emission tomography. Furthermore, the clinical assessment provides the evidence that the neurological phenotype is very slowly progressive. We utilized the 10K single-nucleotide polymorphism (SNP) microarray genotyping for linkage analysis. Genome-wide scan indicated a 45.9-Mb region with a 4.2353 logarithm of the odds score on chromosome 11. Affymetrix genome-wide human SNP array 6.0 assay did not show any gross chromosomal abnormality. Targeted sequencing of two candidate genes within the linkage interval (PAX6 and B3GALTL) as well as mtDNA genome sequencing did not reveal any putative mutations.

Use of hyperbaric oxygen therapy of purpura fulminans in an extremely low birth weight preterm: A case report.

Purpura fulminans (PF) is a rare and fatal complication of septic shock or diffuse intravascular coagulation (DIC) resulting in skin and soft tissue necrosis. PF can be caused by congenital or acquired protein C (PC) or protein S (PS) deficiency. The most common cause of PF in a neonate is sepsis. In our extremely low birth weight preterm case, due to PF that started in the right-hand fingers, examination was made and protein S deficiency was detected as well as MTHFR (A1298C) and Factor V Leiden (R506Q) homozygous mutations. While being unresponsive to fresh frozen plasma (FFP) and unfractionated heparin (UFH) therapy, we want to highlight the curative treatment with hyperbaric oxygen (HBOT), which has not previously been used in extremely low birth weight preterm infants for this purpose.

Multiviral Quartet Nanocages Elicit Broad Anti-Coronavirus Responses for Proactive Vaccinology.

Defending against future pandemics may require vaccine platforms that protect across a range of related pathogens. The presentation of multiple receptor-binding domains (RBDs) from evolutionarily-related viruses on a nanoparticle scaffold elicits a strong antibody response to conserved regions. Here we produce quartets of tandemly-linked RBDs from SARS-like betacoronaviruses coupled to the mi3 nanocage through a SpyTag/SpyCatcher spontaneous reaction. These Quartet Nanocages induce a high level of neutralizing antibodies against several different coronaviruses, including against viruses not represented on the vaccine. In animals primed with SARS-CoV-2 Spike, boost immunizations with Quartet Nanocages increased the strength and breadth of an otherwise narrow immune response. Quartet Nanocages are a strategy with potential to confer heterotypic protection against emergent zoonotic coronavirus pathogens and facilitate proactive pandemic protection.

Novel mutation in GLRB in a large family with hereditary hyperekplexia.

Hereditary hyperekplexia (HH) is a disorder of the inhibitory glycinergic neurotransmitter system. Mutations in five genes have been reported to cause the disease. However, only single mutation in GLRB, the gene encoding beta-subunit of the glycine receptor, in a singleton patient with HH has been found to date. In this study, 13 patients with HH were identified through neurology and genetic clinics. Formal clinical examinations, linkage analysis, homozygosity mapping, in-mutation screening of GLRB and in silico functional analyses were carried out. A novel mutation in GLRB among nine patients was identified. This c.596 T>G perturbation results in the change of the highly conserved methionine at position 177 to arginine. Besides the classical HH phenotype, seven patients had esotropia and few of them had behavioral problems. This study presents a large family with HH as a result of homozygous mutation in GLRB and expands the clinical spectrum of HH to include eye misalignment disorder. Moreover, the report of these familial cases supports the previous evidence in a single patient of an autosomal recessive inheritance of HH because of defects in GLRB.

White-light generation using spatially-structured beams of femtosecond radiation.

We studied white-light generation in water using spatially- structured beams of femtosecond radiation. By changing the transverse spatial phase of an initial Gaussian beam with a 1D spatial light modulator to that of an Hermite-Gaussian (HGn,m) mode, we were able to generate beams exhibiting phase discontinuities and steeper intensity gradients. When the spatial phase of an initial Gaussian beam (showing no significant white-light generation) was changed to that of a HG01, or HG11 mode, significant amounts of white-light were produced. Because self-focusing is known to play an important role in white-light generation, the self-focusing lengths of the resulting transverse intensity profiles were used to qualitatively explain this production. Distributions of the laser intensity for beams having step-wise spatial phase variations were modeled using the Fresnel-Kirchhoff integral in the Fresnel approximation and found to be in good agreement with experiment.

ESCRT recruitment to mRNA-encoded SARS-CoV-2 spike induces virus-like particles and enhanced antibody responses.

Prime-boost regimens for COVID-19 vaccines elicit poor antibody responses against Omicron-based variants and employ frequent boosters to maintain antibody levels. We present a natural infection-mimicking technology that combines features of mRNA- and protein nanoparticle-based vaccines through encoding self-assembling enveloped virus-like particles (eVLPs). eVLP assembly is achieved by inserting an ESCRT- and ALIX-binding region (EABR) into the SARS-CoV-2 spike cytoplasmic tail, which recruits ESCRT proteins to induce eVLP budding from cells. Purified spike-EABR eVLPs presented densely-arrayed spikes and elicited potent antibody responses in mice. Two immunizations with mRNA-LNP encoding spike-EABR elicited potent CD8+ T-cell responses and superior neutralizing antibody responses against original and variant SARS-CoV-2 compared to conventional spike-encoding mRNA-LNP and purified spike-EABR eVLPs, improving neutralizing titers >10-fold against Omicron-based variants for three months post-boost. Thus, EABR technology enhances potency and breadth of vaccine-induced responses through antigen presentation on cell surfaces and eVLPs, enabling longer-lasting protection against SARS-CoV-2 and other viruses.

Novel intragenic deletion in OPHN1 in a family causing XLMR with cerebellar hypoplasia and distinctive facial appearance.

X-linked mental retardation (XLMR) is notably a heterogeneous condition and often poses a diagnostic challenge. The oligophrenin 1 gene (OPHN1) is a protein with a Rho-GTPase-activating domain required in the regulation of the G-protein cycle. Mutations in the OPHN1 cause XLMR with cerebellar hypoplasia and distinctive facial appearance. We report a large Saudi family of four boys and one girl affected with XLMR. The boys had moderate MR, seizure disorder, facial dysmorphism, and cerebellar vermis hypoplasia. The girl had mild MR, seizures, and mild cerebellar hypoplasia. A novel deletion of at least exons 7-15 was identified by polymerase chain reaction analysis and multiple ligation probe amplification of the OPHN1 gene. The array comparative genomic hybridization further delineated approximately 68 kb deletion of the 7-15 exons and nearly half of intron 15. In addition, the X-inactivation confirmed random pattern in the girl. Although the affected boys have remarkably similar phenotype, there was some variability in the severity of the seizure disorder and the cerebellar hypoplasia. The report confirms the previous findings that carrier females may be symptomatic.

In situ tomography of femtosecond optical beams with a holographic knife-edge.

We present an in situ beam characterization technique to analyze femtosecond optical beams in a folded version of a 2f-2f setup. This technique makes use of a two-dimensional spatial light modulator (SLM) to holographically redirect radiation between different diffraction orders. This manipulation of light between diffraction orders is carried out locally within the beam. Because SLMs can withstand intensities of up to I ∼ 10(11) W/cm2, this makes them suitable for amplified femtosecond radiation. The flexibility of the SLM was demonstrated by producing a diverse assortment of "soft apertures" that are mechanically difficult or impossible to reproduce. We test our method by holographically knife-edging and tomographically reconstructing both continuous wave and broadband radiation in transverse optical modes.

Insight into the Temperature Evolution of Electronic Structure and Mechanism of Exchange Interaction in EuS.

Discovered in 1962, the divalent ferromagnetic semiconductor EuS (TC = 16.5 K, Eg = 1.65 eV) has remained constantly relevant to the engineering of novel magnetically active interfaces, heterostructures, and multilayer sequences and to combination with topological materials. Because detailed information on the electronic structure of EuS and, in particular, its evolution across TC is not well-represented in the literature but is essential for the development of new functional systems, the present work aims at filling this gap. Our angle-resolved photoemission measurements complemented with first-principles calculations demonstrate how the electronic structure of EuS evolves across a paramagnetic-ferromagnetic transition. Our results emphasize the importance of the strong Eu 4f-S 3p mixing for exchange-magnetic splittings of the sulfur-derived bands as well as coupling between f and d orbitals of neighboring Eu atoms to derive the value of TC accurately. The 4f-3p mixing facilitates the coupling between 4f and 5d orbitals of neighboring Eu atoms, which mainly governs the exchange interaction in EuS.

Variability of serum oxidative stress biomarkers relative to biochemical data and clinical parameters of glaucoma patients.

PURPOSE: The importance of oxidative stress in both the formation and the course of glaucoma has been known. Among the antioxidants, vitamin E possesses the specific effects and regulatory mechanisms of a neurohormone. The serum oxidant/antioxidant profile is reportedly altered in ocular pathologies. In this study, we analyzed the effect of the clinical parameters of glaucoma and biochemical data on antioxidants and serum oxidative stress markers as oxidation degradation products. METHODS: In this multicenter case control study, control and patient groups consisted of 31 healthy individuals and 160 glaucoma patients with no known additional abnormalities, respectively. We analyzed the oxidation degradation products malonyl dialdehyde (MDA), advanced oxidation protein products (AOPP), antioxidants, vitamins E and A, Serine (Ser), superoxide dismutase (SOD), glutathione peroxidase (Gpx), transferrine (TF), and total antioxidant capacity (TADA). All of these parameters and their relationships with serum cholesterol, glucose, protein, albumin, triglyceride levels, age, gender, visual acuities, intraocular pressure (IOP), c/d ratio, gonioscopic findings, medications, presence of pseudoexfoliation (px), central visual field and Optical Coherence Tomography (OCT) data, pachymetry, and Laplace values, were evaluated individually. Statistical comparisons were performed among them, and with the control group as well. RESULTS: TADA, AOPP, SOD, and Gpx were found to be decreased, and MDA, Ser, TF, vitamins A and E increased in the patient group. All data, excluding AOPP, varied significantly. Vitamin E was the most consistent parameter. CONCLUSIONS: In this study, the association between glaucoma and lipid oxidation was shown on a systematic basis, and the significance of vitamin E as a neuroprotective agent has been revealed once more.

Is the Behcet's disease a risk factor for osteoporosis and is relation to cytokines?

BACKGROUND AND OBJECTIVES: To determine if the Behcet's disease poses a risk for a lower bone mineral density (BMD), and the relation to cytokines. Behcet's disease is a complex, multisystemic, chronic inflammatory disorder. METHODS: This study was carried out on 60 patients and 24 sex- and age-matched healthy controls. Serum osteocalcin (OK) and bone specific alkaline phosphatase (BALP), which are bone formation markers and urine deoxipyridinolin (DOP) levels that is bone resorption marker were evaluated in patient and controls groups. Serum IL-1 and TNF-alpha levels were measured in both groups. BMD was measured with dual X-ray absorptiometry (DEXA) at the lumbar spine and left femur. RESULTS: There were no significant differences in OK and DOP levels between BD and control groups. BALP levels were significantly higher in BD than control groups (p = 0.002). Although there were no statistically significant differences in IL-1 levels between BD and control groups, TNF-alpha levels were significantly higher in BD than control groups. There were significant differences in BMD values of lumbar spine (p < 0.05). No significant differences in BMD values of the left femur were detected in the groups (p > 0.05). CONCLUSIONS: Behcet's disease can be a risk for osteoporosis especially in the lumbar spine. We determined a negative correlation between IL-1 and femur neck BMD (Tab. 4, Fig. 2, Ref. 23).

Precisely Alternating Functionalized Polyampholytes Prepared in a Single Pot from Sustainable Thiolactone Building Blocks.

Polyampholytes with precisely alternating cationic and anionic functional groups were prepared using sustainable thiolactone building blocks in a simple one-pot procedure at room temperature and in water. Ring opening of the N-maleamic acid-functionalized homocysteine thiolactone monomer enabled the introduction of different functional groups into the polymer chain, which contributed to both ionic and hydrogen bonding interactions. The resulting polyampholytes exhibited various isoelectric points while maintaining high solubility in water under different pH and ionic strengths, which expands their potential applications. Finally, it is shown that the upper critical solution temperature (UCST) of these alternating polyampholytes in water/ethanol (30/70% vol) solutions can be tuned as a function of the content of ionic and hydroxyl groups.