Usefulness of arterial spin labeling imaging, which contributed to the early detection of cerebellitis complicated by clinically mild encephalitis/encephalopathy with a reversible splenial lesion: Lessons from three cases.

BACKGROUND: Cerebellitis is a rare complication of clinically mild encephalitis/encephalopathy with a reversible splenial lesion (MERS); however, MERS with cerebellitis is associated with a higher risk of neurological sequelae in comparison to MERS alone. Although the disease is difficult to diagnose by conventional MRI in the early disease phase, arterial spin labeling (ASL), a noninvasive MRI perfusion technique using magnetically-labeled arterial blood water protons, is considered promising. CASE REPORT: We experienced three cases of MERS with cerebellitis. Diffusion-weighted imaging showed a high-intensity lesion at the splenium of the corpus callosum. ASL showed increased blood flow in the cerebellum in all three cases, despite cerebellar symptoms being inapparent or difficult to notice in the early phase of disease in all cases. Patients received methylprednisolone pulse therapy and intravenous immunoglobulin from the early phase of the disease and recovered without neurological sequelae. DISCUSSION: ASL magnetic response imaging simultaneously showed an area of hyperperfusion in the cerebellum. At the same time, the apparent diffusion coefficient of the splenial lesion was decreased in all three cases. The successful diagnosis of cerebellitis in the acute phase led to early therapeutic intervention, which may be important for this condition. We report the usefulness of ASL and review the relevant literature on MERS with cerebellitis.

Predicting the Onset of Acute Encephalopathy With Biphasic Seizures and Late Reduced Diffusion by Using Early Laboratory Data.

Background: Acute encephalopathy with biphasic seizures and late reduced diffusion (AESD) often causes various neurological sequelae, necessitating early and objective differentiation of AESD from a febrile seizure (FS). Therefore, we developed a scoring system that predicts AESD onset using only early laboratory data. Methods: We selected patients with AESD or FS admitted to the Tottori University Hospital between November 2005 and September 2020 and collected laboratory data from onset to discharge in patients with FS and from onset to the second neurological events in patients with AESD. Results: We identified 18 patients with AESD and 181 patients with FS. In comparison with patients with FS, patients with AESD showed statistically significant increases in ammonia (NH3), blood sugar (BS), and serum creatinine (Cr) levels, and the white blood cell (WBC) count, and a significant decrease in pH at <3 h from onset. We set the cut-off values and adjusted the weight of each of these parameters based on data obtained <3 h from onset and proposed a scoring system for predicting AESD. This system showed 91% sensitivity and 94% specificity for distinguishing AESD from FS. These accuracies were only slightly improved by the addition of information related to consciousness and seizure duration (sensitivity, 91%; specificity, 96%). Conclusion: NH3, BS, and Cr levels, WBC count, and pH were significantly different between patients with AESD and patients with FS at <3 h from seizure onset. This scoring system using these data may enable the prediction of AESD onset for patients under sedation or without precise clinical information.

Further characterization of CAPOS/CAOS syndrome with the Glu818Lys mutation in the ATP1A3 gene: A case report.

A 38-year-old female patient experienced recurrent episodes of neurological deterioration during febrile illness at the age of 7 and 8 months, and 2, 4, and 37 years. Acute symptoms comprised unconsciousness, headache, abnormal ocular movements, flaccid paralysis with areflexia, ataxia, dysphagia, and movement disorders. Each episode of neurological deterioration was followed by partial recovery with residual symptoms of progressive disturbance of visual acuity with optic atrophy and hearing loss, moderate intellectual disability, strabismus, ophthalmoplegia, as well as fluctuating degree of gait ataxia, chorea, tremor, and myoclonus. In addition, electrocardiography revealed incomplete right bundle branch block. The genetic testing revealed a de novo heterozygous mutation of c.2452G > A (p.Glu818Lys) in the ATP1A3 gene, which was compatible with the clinical phenotype of CAPOS (cerebellar ataxia, areflexia, pes cavus, optic atrophy, and sensorineural hearing loss)/CAOS syndrome. Here we discuss the significance of clinical features of a patient, overlapping with those of alternating hemiplegia of childhood, along with a literature review.

CACNA1A-related early-onset encephalopathy with myoclonic epilepsy: A case report.

We report a one-year-old boy with early-onset myoclonic epilepsy, developmental arrest, and hyperekplexia during early infancy. He presented with refractory myoclonic/tonic seizures since birth. Electroencephalography revealed multifocal spikes, and rhythmic activities that occurred simultaneous with aggravation of myoclonus accompanied by tonic upper limb elevation. Brain magnetic resonance imaging revealed progressive cerebral atrophy with periventricular signal change and thin corpus callosum at one year of age. A de novo heterozygous missense mutation in the CACNA1A gene was confirmed. This patient was the most severe phenotype of CACNA1A-related early-onset encephalopathy among previous reports.

Quantification of DNA fragmentation in processed foods using real-time PCR.

DNA analysis of processed foods is performed widely to detect various targets, such as genetically modified organisms (GMOs). Food processing often causes DNA fragmentation, which consequently affects the results of PCR analysis. In order to assess the effects of DNA fragmentation on the reliability of PCR analysis, we investigated a novel methodology to quantify the degree of DNA fragmentation. We designed four real-time PCR assays that amplified 18S ribosomal RNA gene sequences common to various plants at lengths of approximately 100, 200, 400, and 800 base pairs (bp). Then, we created an indicator value, "DNA fragmentation index (DFI)", which is calculated from the Cq values derived from the real-time PCR assays. Finally, we demonstrated the efficacy of this method for the quality control of GMO detection in processed foods by evaluating the relationship between the DFI and the limit of detection.

Atomic Force Microscopy of DNA-wrapped Single-walled Carbon Nanotubes in Aqueous Solution.

We evaluated hybrids of DNA and single-walled carbon nanotubes (SWNTs) in aqueous solution and in air using atomic force microscopy (AFM). Although intensive AFM observations of these hybrids were previously carried out for samples in air, this is the first report on AFM observations of these hybrids in solution. As expected, diameters of DNA-SWNT hybrids dramatically increased in tris(hydroxymethyl)aminomethane-ethylenediaminetetraacetic acid (TE) buffer solution. The data suggest that DNA molecules maintain their structures even on the SWNT surfaces. Furthermore, we simultaneously observed single DNA-SWNT hybrids using three different AFM modes in air and in the TE buffer solution. Height value of the hybrids was largest in the solution, and lowest for the mode that repulsive force is expected in air. For the bare SWNT molecules, height differences among the three AFM modes were much lower than those of the DNA-SWNT hybrids. DNA molecules adsorbed on SWNT surfaces flexibly changed their morphology as well as DNA molecules on flat surfaces such as mica. This is hopeful results for biological applications of DNA-SWNT hybrids. In addition, our results revealed the importance of the single-molecule approach to evaluate DNA structures on SWNT surfaces.

Selective binding of single-stranded DNA-binding proteins onto DNA molecules adsorbed on single-walled carbon nanotubes.

Single-stranded DNA-binding (SSB) proteins were treated with hybrids of DNA and single-walled carbon nanotubes (SWNTs) to examine the biological function of the DNA molecules adsorbed on the SWNT surface. When single-stranded DNA (ssDNA) was used for the hybridization, significant binding of the SSB molecules to the ssDNA-SWNT hybrids was observed by using atomic force microscopy (AFM) and agarose gel electrophoresis. When double-stranded DNA (dsDNA) was used, the SSB molecules did not bind to the dsDNA-SWNT hybrids in most of the conditions that we evaluated. A specifically modified electrophoresis procedure was used to monitor the locations of the DNA, SSB, and SWNT molecules. Our results clearly showed that ssDNA/dsDNA molecules on the SWNT surfaces retained their single-stranded/double-stranded structures.

Controlling the adsorption and desorption of double-stranded DNA on functionalized carbon nanotube surface.

We demonstrated controlled adsorption and desorption of double-stranded DNA (dsDNA) on single-walled carbon nanotube (SWNT) surface functionalized with polyethyleneglycol (PEG SWNT). First, when dsDNA molecules were mixed with the PEG SWNT solution, the DNA molecules spontaneously adsorbed onto the PEG SWNT surface and formed dsDNA-PEG SWNT conjugates without sonication. Next, we succeeded in detaching the dsDNA adsorbed on PEG SWNT by annealing at 95°C for 30 min. These results were confirmed using atomic force microscopy, agarose gel electrophoresis, and micro-Raman spectroscopy. In contrast, when we used the usual SWNT produced by the high-pressure carbon monoxide method (HiPco SWNT), the DNA molecules were fragmented during the adsorption process as sonication was necessary for the hybridization of DNA-SWNT conjugates. Furthermore, detachment of DNA molecules from HiPco SWNT by annealing was impossible. Our method may be useful for developing DNA devices using SWNTs as substrates when it is combined with previously established various biochemical techniques.

Surface morphology of hybrids of double-stranded DNA and single-walled carbon nanotubes studied by atomic force microscopy.

We examined the formation of hybrids of double-stranded DNA (dsDNA) and single-walled carbon nanotubes (SWNTs), which has not been well investigated yet. In particular, the adsorption of dsDNA onto SWNT produced by chemical vapor deposition (CVD) was examined for the first time. When small amount of dsDNA was mixed with CVD SWNT, well dispersed hybrids with smooth surfaces were observed by atomic force microscopy (AFM). Through a comparison of dsDNA, single-stranded DNA (ssDNA), CVD SWNT, and high-pressure carbon monoxide process (HiPco) SWNT, we found that the surface morphology of the hybrids was independent of the DNA type. Even when sonicated salmon testes DNA, which has a random sequence and length, was employed, smooth surfaces were obtained on the dsDNA-CVD hybrids as well as on the ssDNA-CVD hybrids. The ratio of monodispersed SWNT and bundled SWNT in a dispersion solution was also not affected by the DNA type. In contrast, the quantity of the fabricated hybrids was affected by the types of DNA especially when HiPco SWNT was used. Our results indicated that characteristic features of the dsDNA-CVD hybrids and provide an enhanced understanding of the adsorption mechanism of dsDNA onto SWNTs.

A case of nonfunctioning pituitary carcinoma that responded to temozolomide treatment.

Pituitary carcinoma is a rare malignancy and is difficult to manage. Pituitary carcinomas commonly produce either PRL or ACTH, but some do not produce pituitary hormones. The alkylating reagent temozolomide (TMZ) was recently shown to be effective as a treatment for pituitary carcinoma. Most of the published reports of TMZ use in pituitary carcinoma cases were against hormone-producing carcinomas. Only a few patients with a nonfunctioning pituitary carcinoma treated with TMZ have been reported. Here we describe our treatment of a patient with nonfunctioning pituitary carcinoma and a background of multiple endocrine neoplasia type 1. The pituitary carcinoma was accompanied by meningeal dissemination with cerebral and L1 spinal bone metastasis. The patient received continuous dosing of TMZ along with external radiation, followed by standard dosing of TMZ. There was an apparent antitumor response seen in MRI. MGMT, an enzyme antagonized by TMZ, was negative in the tumor. The therapeutic efficacy of TMZ and dosing schedules of TMZ in pituitary carcinoma are discussed.

Effect of embryo transfer following artificial insemination (ETFAI) on reproductive performance in dairy cows in South-Western Japan.

In order to improve the reproductive performance during the summer period, embryo transfer (ET; Japanese black embryo) following artificial insemination (AI; Holstein semen: ETFAI) was conducted in dairy cows in south-western Japan (n=56). The conception rate was improved in cows with ETFAI compared with conventional AI, which served as the control (n=195; 30.4% vs. 13.8%, P<0.01). However, higher fetal loss was observed in ETFAI compared with the controls (38.1% vs. 7.4%, P<0.05). Four cases of twin pregnancy resulted in 2 singletons and a set of twins. There was no difference in the plasma progesterone level on d0 or d7 (d0=AI), but rather lower rectal temperature was observed on d7 or d8 (38.7 degrees C vs. 39.4 degrees C and 38.8 degrees C vs. 39.1 degrees C, P<0.05) in pregnant cows compared with those that were open. ETFAI could improve reproductive performance in dairy cows during the summer period in south-western Japan.