[Analysis of perrault syndrome caused by pathogenic variants in LARS2 and HARS2 genes].

Objective: To investigate the molecular etiology of Perrault syndrome by analyzing the clinical phenotype and pathogenic gene variants of 2 male patients with bilateral severe sensorineural deafness. Methods: Two male patients with Perrault syndrome characterized by severe sensonrineual deafness adimitted to the First Affiliated Hospital of Zhengzhou University between February 2021 and March 2022 were selected, and the clinical phenotype and pathogenic gene variants of them and their family members were summarized. The whole exome sequencing technology was used to screen the pathogenic variants of the probands, and the candidate variants were determined by combining with clinical phenotype. The probands and their family members were verified by the Sanger sequencing method. Results: The whole exome sequencing results showed that the proband of family 1 had a compound heterozygous variants of the LARS2 (NM_015340.4) gene c.1565C>A (p.Thr522Asn) and c.1079T>C (p.Ile360Thr). The reported pathogenic variant c.1565C>A came from the mother, and the novel variant c.1079T>C came from the father. The second proband harbored compound heterozygous variants of HARS2 gene (NM_012208.4) c.1273C>T (p.Arg425Trp) and c.1403G>C (p.Gly468Ala), with the former from the proband's mother, the latter from the father. The c.1273C>T was novel and c.1403G>C was the reported pathogenic variant. All above variants were respectively classified as pathogenic, uncertain significance, uncertain significance and likely pathogenic based on the ACMG guidelines. Conclusion: This study expands the mutational spectrum of LARS2 and HARS2 genes, which highlights that genetic testing plays an important role in the early diagnosis of syndromic deafness.

[Branchio-oto-renal syndrome or branchio-oto syndrome: the clinical and genetic analysis in five Chinese families].

Objective: To screen the causative genes of five families with branchio-oto-renal syndrome (BORS) or branchio-oto syndrome(BOS) and to analyze the phenotypic characteristics and clinical management strategies of patients. Methods: Five families with BORS/BOR from December 2018 to September 2021 were recruited, information of patients, including family history and medical history, was collected, and genealogies were drawn. The examinations concerning audiology, nephrology, and radiology were performed on the affected individuals. Peripheral blood was obtained for DNA extraction, then next-generation sequencing technology was used to screen candidate variants associated with BORS/BOS. Based on patient's clinical results, the appropriate interventions were recommended and implemented. Results: Eight individuals were diagnosed with BOS or BORS. Of the eight patients, all had hearing loss, preauricular pits and ear malformations, and only four presented with branchial cleft fistulae or cysts. Except for two patients(5-I-2, 5-II-2) who did not undergo renal examination, the remaining six lacked renal abnormalities. Genetic analysis identified four likely pathogenic or pathogenic EYA1 variants (c.1715G>T, c.1140+1G>A, c.639G>C, c.1475+1G>C; NM_000503.6), and c.1715G>T was first reported in this study. Middle ear ossicular reconstruction was performed in 1-II-2,2-I-2 and 3-II-2, but did not yield the expected results; then hearing aids and cochlear implantation were recommended and achieved satisfactory results. Conclusions: Next-generation sequencing technology facilitates the diagnosis and genetic counseling of BORS/BOS. Hearing loss, preauricular pits, ear malformations and branchial cleft fistulae or cysts are the most common manifestations of patients in this study. Middle ear surgeries for improving hearing loss may have some limitations in BORS/BOS patients, and hearing aids and cochlear implantation can contribute to hearing gains.

[Study on syndromic deafness caused by novel pattern of compound heterozygous variants in the CDH23 gene].

Objective: To explore the pathogenic variants of a family with syndromic deafness by high-throughput sequencing. Methods: The family was from Puyang City, Henan Province, and had four members, including two with syndromic deafness. The proband and his sister had congenital deafness, and their parents had normal phenotypes. The clinical phenotype of the family was characterized using clinical examinations and pedigree analysis. The clinical examinations included imaging examination, audiometry (pure tone audiometry, acoustic immittance, brainstem auditory evoked potential, and otoacoustic emission), vestibular function test, and ophthalmic examination (visual acuity test, visual field test, fundus examination, visual evoked potential, and electroretinogram). Target exome sequencing of 129 known deafness genes and bioinformatics analysis were used to screen suspected pathogenic variants. Sanger sequencing and minigene assay were used to verify and functionally investigate the mutation detected, respectively. According to the standards and guidelines for interpreting genetic variants proposed by the American College of Medical Genetics and Genomics, the variants c.6049G>A and c.8699A>G were classified as pathogenic/likely pathogenic, and the variant c.9856C>G was classified as variants of uncertain significance. Results: The probands and his sister had severe sensorineural hearing loss with decreased binocular vision, night blindness, decreased peripheral visual field sensitivity and partial visual field defect, and normal vestibular function. Both of them had three CDH23 mutations, including CDH23 (NM_022124.5) c.6049G>A (p.Gly2017Ser),c.9856C>G (p.His3286Asp), and c.8699A>G (p. Asp2900Gly), The first two were inherited from the father, and the last one was from the mother. The missense variants c.9856C>G and c.8699A>G were not included in the gnomad database. The missense mutation c.6049G>A was located in the last position of exon 46 and was predicted to affect splicing by bioinformatics software. The minigene experiment showed that the mutation cause exon skipping of exon 46, resulting in an abnormal protein. Conclusions: Compound heterozygous variations of the CDH23 are the leading cause of USH1D in the family. This study confirms that the compound heterozygosity of splicing and missense variants of the CDH23 gene could lead to USH1D.

[The study of AAV9 expression in cochleae of mice at different ages].

Objective: To determine the extent of transfection and expression of adeno-associated virus (AAV) serotype 9 (AAV9) in the cochleae of mice at different ages. Methods: AAV9-green fluorescent protein (GFP) was inoculated into the cochlea of mice via the round window membrane (RWM) or through cochleostomy at different ages. Four groups were divided according to ages and injection sites: P1SM group, AAV9-GFP was delivered to the scala media by cochleostomy at postnatal day 1; P1RW group, AAV9-GFP was delivered to the scala tympani via RWM at postnatal day 1; P9RW group: AAV9-GFP was injected through RWM at postnatal day 9; and P30RW group, adult mice (P30) were injected through RWM. GFP expression in cochlear whole mount was analyzed and auditory brainstem response (ABR) tests were conducted one month after virus injection (for each animal, only left cochlea was injected and the right side was used as a control). GraphPad Prism 5 statistical software was used for data analysis. Results: All of inner hair cells (IHCs) and most of outer hair cells (OHCs) were transfected via two approaches at P1 injection. There was no significant difference in ABR threshold between injected ears and untreated ears (P>0.05). All of the IHCs and parts of OHCs (69% in apical turn) were transfected via RWM at P9. The strongest GFP expression was observed near the apical turn. Cochlear inoculation via RWM at P30 led to transgene expression in only IHCs. The ABR threshold of injected ears in P9RW group and P30RW group was significantly higher than that of contralateral ears (P<0.01). Conclusions: AAV9 can be highly expressed in the inner and outer hair cells of the cochlea and hearing sensitivity can be preserved if virus injections are performed in neonatal mice. After AAV9 is transfected into the inner ear of adult mice, it is only expressed in the inner hair cells, which leads to the increase of the ABR response threshold of mice. Transfection efficiency is significant higher in neonate mice than in P9 and adult mice.

Origin of quantum ring formation during droplet epitaxy.

Droplet epitaxy of GaAs is studied in real time using in situ surface electron microscopy. The resulting movies motivate a theoretical model for quantum ring formation which can explain the origin of nanoscale features such as double rings observed under a variety of experimental conditions. Inner rings correspond to GaAs deposition at the droplet edge, while outer rings result from the reaction of Ga and As atoms diffusing along the surface. The observed variety of morphologies primarily reflects relative changes in the outer rings with temperature and As flux.

Elementary excitations at magnetic surfaces and their spin dependence.

The elementary surface excitations are studied by spin-polarized electron energy loss spectroscopy on a prototype oxide surface [an oxygen passivated Fe(001)-p(1×1) surface], where the various excitations coexist. For the first time, the surface phonons and magnons are measured simultaneously and are distinguished based on their different spin nature. The dispersion relation of all excitations is probed over the entire Brillouin zone. The different phonon modes observed in our experiment are described by means of ab initio calculations.

Caustic imaging of gallium droplets using mirror electron microscopy.

We discuss a new interpretation of mirror electron microscopy (MEM) images, whereby electric field distortions caused by surface topography and/or potential variations are sufficiently large to create caustics in the image contrast. Using a ray-based trajectory method, we consider how a family of rays overlaps to create caustics in the vicinity of the imaging plane of the magnetic objective lens. Such image caustics contain useful information on the surface topography and/or potential, and can be directly related to surface features. Specifically we show how a through-focus series of MEM images can be used to extract the contact angle of a Ga droplet on a GaAs (001) surface.

Decomposition controlled by surface morphology during langmuir evaporation of GaAs.

When GaAs is heated in vacuum, it decomposes into Ga and As as it evaporates. Real-time in situ surface electron microscopy reveals striking bursts of "daughter" droplet nucleation and growth when coalescence of large "parent" droplets exposes nonplanar surface regions. We analyze the behavior, predicting a morphology-dependent congruent evaporation temperature. Based on this we propose a new approach for the self-assembly and positioning of quantum structures via droplet epitaxy, which we demonstrate at the proof-of-concept level.

Asymmetric spin-wave dispersion on Fe(110): direct evidence of the Dzyaloshinskii-Moriya interaction.

The influence of the Dzyaloshinskii-Moriya interaction on the spin-wave dispersion in an Fe double layer grown on W(110) is measured for the first time. It is demonstrated that the Dzyaloshinskii-Moriya interaction breaks the degeneracy of spin waves and leads to an asymmetric spin-wave dispersion relation. An extended Heisenberg spin Hamiltonian is employed to obtain the longitudinal component of the Dzyaloshinskii-Moriya vectors from the experimentally measured energy asymmetry.

Magnons in a ferromagnetic monolayer.

We report the first observation of high wave vector magnon excitations in a ferromagnetic monolayer. Using spin-polarized electron energy loss spectroscopy, we observed the magnon dispersion in one atomic layer (ML) of Fe on W(110) at 120 K. The magnon energies are small in comparison to the bulk and surface Fe(110) excitations. We find an exchange parameter and magnetic anisotropy similar to that from static measurements. Our results are in sharp contrast to theoretical calculations, indicating that the present understanding of magnetism of the ML Fe requires considerable revision.

Running droplets of gallium from evaporation of gallium arsenide.

High-temperature annealing of gallium arsenide in vacuum causes excess evaporation of arsenic, with accumulation of gallium as liquid droplets on the surface. Using real-time in situ surface electron microscopy, we found that these droplets spontaneously run across the crystal surface. Running droplets have been seen in many systems, but they typically require special surface preparation or gradient forces. In contrast, we show that noncongruent evaporation automatically provides a driving force for running droplets. The motion is predicted and observed to slow and stop near a characteristic temperature, with the speed increasing both below and above this temperature. The same behavior is expected to occur during the evaporation of similar III-V semiconductors such as indium arsenide.

A randomized controlled trial of high-permeability haemodialysis against conventional haemodialysis in the treatment of uraemic pruritus.

BACKGROUND: Uraemic pruritus (UP) is one of the most common problems in patients with chronic renal failure. Owing to the complexity of UP, no specific treatment is currently available. Recently, the accumulated toxins of mid and macro molecules in advanced renal failure have been proposed to play an important role in the mediation of pruritus. AIM: To evaluate the effect of high permeability haemodialysis (HPHD) against conventional haemodialysis (CHD) on UP. METHODS: A randomized, prospective, double-blind study was performed to compare the efficacy of HPHD against CHD in the treatment of UP. In total, 116 patients with chronic renal failure and UP were enrolled in the trial. A visual analogue scale (VAS) was used to assess the severity of itch. The toxins of mid and macro molecules [beta2-microglobulin (beta2-MG), parathyroid hormone (PTH), respectively] were measured, and the solute clearance rate (SCR) and urea clearance index (Kt/V) were also determined. RESULTS: The pruritus scores in the HPHD group were significantly lower (2.23 +/- 1.05) than those in the CHD group (5.45 +/- 1.91, P = 0.012), although the SCR and Kt/V showed no significance between the two groups (SCR P = 0.075; Kt/V P = 0.082). It was found that HPHD and CHD achieved a reasonable clearance rate of blood urea nitrogen and creatinine. However, the toxins of mid and macro molecules were markedly reduced in the blood of patients treated with HPHD, compared with those treated with CHD. The concentrations of PTH and beta2-MG were significantly reduced by HPHD in comparison with CHD (PTH 119.27 +/- 8.41 vs. 165.18 +/- 9.37 pmol/L, P = 0.01; beta2-MG 3.39 +/- 0.76 vs. 5.92+/- 1.58 g/mL, P = 0.012). CONCLUSIONS: These data indicate that HPHD can efficiently relieve UP through clearance of accumulated mid and macro molecules in vivo. This further supports the hypothesis that these molecules are involved in UP.

Ga droplet morphology on GaAs(001) studied by Lloyd's mirror photoemission electron microscopy.

We apply Lloyd's mirror photoemission electron microscopy (PEEM) to study the surface shape of Ga droplets on GaAs(001). An unusual rectangular-based droplet shape is identified and the contact angle is determined in situ. It is shown that quenching does not appreciably affect droplet shape and ex situ measurements of the contact angle by atomic force microscopy are in good agreement with Lloyd's mirror PEEM. Extension of Lloyd's mirror technique to reconstruct general three-dimensional (3D) surface shapes and the potential use of synchrotron radiation to improve vertical resolution is discussed.

Large wave vector spin waves and dispersion in two monolayer fe on w(110).

We present the first surface spin-wave (SW) dispersion measurements up to the surface Brillouin zone boundary of a two monolayer Fe film on W(110) by using spin-polarized electron energy loss spectroscopy. Pronounced features of SW peaks are observed in the spectra at room temperature. We found that the SW energies in the Fe film are strongly reduced compared to spin waves in bulk Fe and to theoretical predictions. Our results suggest that this reduction is caused by the reduction of exchange interaction within the 2 ML Fe on W(110) as compared to bulk Fe.

Controlled release of insulin from PLGA nanoparticles embedded within PVA hydrogels.

A simple and versatile delivery platform for peptide and protein based on physically cross-linked poly (vinyl alcohol) (PVA) hydrogels containing insulin-loaded poly (lactic-co-glycolic acid) (PLGA) nanoparticles was successfully fabricated. The particle morphology and size were characterized by SEM and laser light scattering method, respectively. Results showed that these particles had a mean diameter of 615 nm with a narrow size distribution and homogeneous particle production. The protein encapsulation efficiency was 72.6%. When insulin-loaded PLGA nanoparticles were administered intraperitoneally as a single dose (20 U/kg) to streptozotocin-induced diabetic mouse, blood glucose levels of these mice decreased and it could be sustained at such levels over 24 h. In vitro release further indicated that entrapment of the nanoparticles into the PVA hydrogels causes a reduction in both the release rate and the total amount of insulin released, which suggesting that PLGA nanoparticles entrapped into the PVA hydrogels showed more suitable controlled release kinetics for protein delivery.

Body-centered-cubic Ni and its magnetic properties.

The body-centered-cubic (bcc) phase of Ni, which does not exist in nature, has been achieved as a thin film on GaAs(001) at 170 K via molecular beam epitaxy. The bcc Ni is ferromagnetic with a Curie temperature of 456 K and possesses a magnetic moment of 0.52+/-0.08 micro(B)/atom. The cubic magnetocrystalline anisotropy of bcc Ni is determined to be +4.0x10(5) ergs x cm(-3), as opposed to -5.7x10(4) ergs x cm(-3) for the naturally occurring face-centered-cubic (fcc) Ni. This sharp contrast in the magnetic anisotropy is attributed to the different electronic band structures between bcc Ni and fcc Ni, which are determined using angle-resolved photoemission with synchrotron radiation.

Localization of pigment epithelium derived factor (PEDF) in developing and adult human ocular tissues.

PURPOSE: To localize pigment epithelium-derived factor (PEDF) in developing and adult human ocular tissues. METHODS: PEDF was localized in fetal and adult eyes by immunofluorescence with a polyclonal antibody (pAb) against amino acids 327-343 of PEDF, or a monoclonal antibody (mAb) against the C-terminal 155 amino acids of PEDF. Specificity of the antibodies was documented by Western blotting. PEDF mRNA was localized in adult retina by in situ hybridization. RESULTS: In developing retinas (7.4 to 21.5 fetal weeks, Fwks), pAb anti-PEDF labeled retinal pigment epithelium (RPE) granules, developing cones, some neuroblasts and many cells in the ganglion cell layer (GCL). In adult retinas, pAb anti-PEDF labeled rod and cone cytoplasm and nuclei of rods but not cones. Cells in the INL and GCL, choroid, corneal epithelium and endothelium, and ciliary body were also pAb PEDF-positive. Preadsorption of pAb anti-PEDF with the immunizing peptide blocked specific labeling in retina and other tissues, except for photoreceptor outer segments. In agreement with the immunolocalization with pAb anti-PEDF, in situ hybridization revealed PEDF mRNA in the RPE, photoreceptors, inner nuclear layer cells and ganglion cells in adult retina. In developing retinas 18 Fwks and older, and in adult retinas, mAb anti-PEDF labeled the interphotoreceptor matrix (IPM). Western blots of retina, cornea, and ciliary body/iris with pAb anti-PEDF produced several bands at about 46 kDa. With mAb anti-PEDF, retina produced one band at about 46 kDa; cornea and ciliary body/iris had several bands at about 46 kDa. CONCLUSIONS: PEDF, originally reported as a product of RPE cells, is present in photoreceptors and inner retinal cell types in developing and adult human eyes. Photoreceptors and RPE may secrete PEDF into the IPM.

cADP-ribose activates reconstituted ryanodine receptors from coronary arterial smooth muscle.

The present study was designed to test the hypothesis that cADP-ribose (cADPR) increases Ca(2+) release through activation of ryanodine receptors (RYR) on the sarcoplasmic reticulum (SR) in coronary arterial smooth muscle cells (CASMCs). We reconstituted RYR from the SR of CASMCs into planar lipid bilayers and examined the effect of cADPR on the activity of these Ca(2+) release channels. In a symmetrical cesium methanesulfonate configuration, a 245 pS Cs(+) current was recorded. This current was characterized by the formation of a subconductance and increase in the open probability (NP(o)) of the channels in the presence of ryanodine (0.01-1 microM) and imperatoxin A (100 nM). A high concentration of ryanodine (50 microM) and ruthenium red (40-80 microM) substantially inhibited the activity of RYR/Ca(2+) release channels. Caffeine (0.5-5 mM) markedly increased the NP(o) of these Ca(2+) release channels of the SR, but D-myo-inositol 1,4,5-trisphospate and heparin were without effect. Cyclic ADPR significantly increased the NP(o) of these Ca(2+) release channels of SR in a concentration-dependent manner. Addition of cADPR (0.01 microM) into the cis bath solution produced a 2.9-fold increase in the NP(o) of these RYR/Ca(2+) release channels. An eightfold increase in the NP(o) of the RYR/Ca(2+) release channels (0.0056 +/- 0.001 vs. 0.048 +/- 0.017) was observed at a concentration of cADPR of 1 microM. The effect of cADPR was completely abolished by ryanodine (50 microM). In the presence of cADPR, Ca(2+)-induced activation of these channels was markedly enhanced. These results provide evidence that cADPR activates RYR/Ca(2+) release channels on the SR of CASMCs. It is concluded that cADPR stimulates Ca(2+) release through the activation of RYRs on the SR of these smooth mucle cells.

Concentric retinitis pigmentosa: clinicopathologic correlations.

Progressive concentric (centripetal) loss of vision is one pattern of visual field loss in retinitis pigmentosa. This study provides the first clinicopathologic correlations for this form of retinitis pigmentosa. A family with autosomal dominant concentric retinitis pigmentosa was examined clinically and with visual function tests. A post-mortem eye of an affected 94 year old family member was processed for histopathology and immunocytochemistry with retinal cell specific antibodies. Unrelated simplex/multiplex patients with concentric retinitis pigmentosa were also examined. Affected family members of the eye donor and patients from the other families had prominent peripheral pigmentary retinopathy with more normal appearing central retina, good visual acuity, concentric field loss, normal or near normal rod and cone sensitivity within the preserved visual field, and reduced rod and cone electroretinograms. The eye donor, at age 90, had good acuity and function in a central island. Grossly, the central region of the donor retina appeared thinned but otherwise normal, while the far periphery contained heavy bone spicule pigment. Microscopically the central retina showed photoreceptor outer segment shortening and some photoreceptor cell loss. The mid periphery had a sharp line of demarcation where more central photoreceptors were near normal except for very short outer segments and peripheral photoreceptors were absent. Rods and cones showed abrupt loss of outer segments and cell death at this interface. It is concluded that concentric retinitis pigmentosa is a rare but recognizable phenotype with slowly progressive photoreceptor death from the far periphery toward the central retina. The disease is retina-wide but shows regional variation in severity of degeneration; photoreceptor death is severe in the peripheral retina with an abrupt edge between viable and degenerate photoreceptors. Peripheral to central gradients of unknown retinal molecule(s) may be defective or modify photoreceptor degeneration in concentric retinitis pigmentosa.

A new bimetallic assembly magnet [[Ni(tn)2]5[FeIII(CN)6]3]n(ClO4)n.2.5nH2O (tn = trimethylenediamine) with a novel 3-D tunnel structure.

A novel molecular-based magnet of three-dimensional (3-D) cyanide-bridged bimetallic assembly, [[Ni(tn)2]5[FeII(CN)6]3]n(ClO4)n.2.5nH2O where tn = trimethylenediamine, was synthesized and structurally characterized. The compound with an asymmetric unit of C24H52.5Cl0.5Fe1.5N19Ni2.5O3.255 crystallized in the monoclinic system of the space group P2(1)/n with a = 10.173(3) A, b = 16.053(2) A, c = 26.309(3) A, beta = 91.30(2) degrees, and Z = 4. The assembly has a 3-D network structure extended by three different types of FeIII-CN-NiII-NC-FeIII linkages. The iron (FeIII) atoms are located in two different chemical environments, which were confirmed by Mössbauer experimental results. The nickel (NiII) atoms have three different coordination environments. Cryomagnetic properties revealed that 3-D magnetic ordering occurs over the lattice below the Curie temperature around 10 K.