Age- and cell cycle-related expression patterns of transcription factors and cell cycle regulators in Müller glia.

Mammalian Müller glia express transcription factors and cell cycle regulators essential for the function of retinal progenitors, indicating the latent neurogenic capacity; however, the role of these regulators remains unclear. To gain insights into the role of these regulators in Müller glia, we analyzed expression of transcription factors (Pax6, Vsx2 and Nfia) and cell cycle regulators (cyclin D1 and D3) in rodent Müller glia, focusing on their age- and cell cycle-related expression patterns. Expression of Pax6, Vsx2, Nfia and cyclin D3, but not cyclin D1, increased in Müller glia during development. Photoreceptor injury induced cell cycle-associated increase of Vsx2 and cyclin D1, but not Pax6, Nfia, and cyclin D3. In dissociated cultures, cell cycle-associated increase of Pax6 and Vsx2 was observed in Müller glia from P10 mice but not from P21 mice. Nfia levels were highly correlated with EdU incorporation suggesting their activation during S phase progression. Cyclin D1 and D3 were transiently upregulated in G1 phase but downregulated after S phase entry. Our findings revealed previously unknown links between cell cycle progression and regulator protein expression, which likely affect the cell fate decision of proliferating Müller glia.

Genotype-phenotype correlation of renal lesions in the tuberous sclerosis complex.

Tuberous sclerosis complex (TSC) is an autosomal dominant disease caused by loss-of-function mutations in either of two tumor suppressor genes, TSC1 and TSC2. These mutations lead to the growth of benign tumors and hamartomas in many organs, including those of the central nervous system, the skin, and the kidneys. To investigate the genotype-phenotype correlation, we performed sequence analysis of the TSC1/2 genes using next-generation sequencing. We classified 30 patients with TSC whose pathogenic variants were identified into two groups: those with mutations producing premature termination codons (PTCs) and those with missense mutations. Then, we compared the phenotypes between the two groups. Patients with a PTC were significantly more likely to manifest the major symptoms of the diagnostic criteria than those without a PTC (P = 0.035). The frequencies of subependymal nodules (P = 0.026), cortical tubers (P = 0.026), and renal cysts (P = 0.026) were significantly higher in PTC-containing variants than in cases without a PTC. When the analyses were limited to renal angiomyolipoma (AML) cases with TSC2 mutations, there was no difference in tumor size between cases with and without a PTC. However, the cases with a PTC showed a trend toward disease onset at a younger age and multiple tumors, and bilateral disease was observed in their AML lesions. TSC patients with PTC-producing mutations might potentially manifest more severe TSC phenotypes than those with missense mutations. A larger-scale study with appropriate samples deserves further investigation.

A case of a parthenogenetic 46,XX/46,XY chimera presenting ambiguous genitalia.

Sex-chromosome discordant chimerism (XX/XY chimerism) is a rare chromosomal disorder in humans. We report a boy with ambiguous genitalia and hypospadias, showing 46,XY[26]/46,XX[4] in peripheral blood cells. To clarify the mechanism of how this chimerism took place, we carried out whole-genome genotyping using a SNP array and microsatellite analysis. The B-allele frequency of the SNP array showed a mixture of three and five allele combinations, which excluded mosaicism but not chimerism, and suggested the fusion of two embryos or a shared parental haplotype between the two parental cells. All microsatellite markers showed a single maternal allele. From these results, we concluded that this XX/XY chimera is composed of two different paternal alleles and a single duplicated maternal genome. This XX/XY chimera likely arose from a diploid maternal cell that was formed via endoduplication of the maternal genome just before fertilization, being fertilized with both X and Y sperm.

Novel mutation in the KITLG gene in familial progressive hyperpigmentation with or without hypopigmentation.

We herein report a novel mutation in familial progressive hyper- and hypopigmentation (FPHH). The KITLG gene encoding the KIT ligand protein is a disease-causing gene for FPHH. Various disease-causing gain-of-function mutations, which reside within or adjacent to the conserved VTNN motif of this gene, have been described to date. We have now identified a novel KITLG mutation, c.337G>A (p.Glu113Lys), in FPHH which is located within another ligand-receptor interaction site.

A female patient with retinoblastoma and severe intellectual disability carrying an X;13 balanced translocation without rearrangement in the RB1 gene: a case report.

BACKGROUND: Female carriers of a balanced X; autosome translocation generally undergo selective inactivation of the normal X chromosome. This is because inactivation of critical genes within the autosomal region of the derivative translocation chromosome would compromise cellular function. We here report a female patient with bilateral retinoblastoma and a severe intellectual disability who carries a reciprocal X-autosomal translocation. CASE PRESENTATION: Cytogenetic and molecular analyses, a HUMARA (Human androgen receptor) assay, and methylation specific PCR (MSP) and bisulfite sequencing were performed using peripheral blood samples from the patient. The patient's karyotype was 46,X,t(X;13)(q28;q14.1) by G-banding analysis. Further cytogenetic analysis located the entire RB1 gene and its regulatory region on der(X) with no translocation disruption. The X-inactivation pattern in the peripheral blood was highly skewed but not completely selected. MSP and deep sequencing of bisulfite-treated DNA revealed that an extensive 13q region, including the RB1 promoter, was unusually methylated in a subset of cells. CONCLUSIONS: The der(X) region harboring the RB1 gene was inactivated in a subset of somatic cells, including the retinal cells, in the patient subject which acted as the first hit in the development of her retinoblastoma. In addition, the patient's intellectual disability may be attributable to the inactivation of the der(X), leading to a 13q deletion syndrome-like phenotype, or to an active X-linked gene on der (13) leading to Xq28 functional disomy.

Prenatal diagnosis of premature chromatid separation/mosaic variegated aneuploidy (PCS/MVA) syndrome.

Premature chromatid separation/mosaic variegated aneuploidy (PCS/MVA) syndrome is a rare genetic disorder. In this case report, we describe the prenatal diagnosis of PCS/MVA syndrome in a 24-year-old, gravida 1, para 1, woman who was referred to us in her second trimester due to fetal growth restriction and extreme microcephaly (-5.0 standard deviations). Amniocentesis and chromosomal analysis confirmed PCS in 80% of cultured fetal cells. PCS findings were positive in 9% of paternal cells and 11% of maternal cells, indicative that both were PCS carriers. Genetic analysis confirmed that the fetus carried a combined heterozygote of maternal G > A point mutation of the promoter area of the BUB1B gene and a paternal Alu sequence insertion between intron 8 and exon 9 of the BUB1B gene. As PCS/MVA syndrome is associated with the development of various malignancies in early life, prenatal diagnosis is important for effective planning of post-natal care.

PCS/MVA syndrome caused by an Alu insertion in the BUB1B gene.

We report a case of premature chromatid separation/mosaic variegated aneuploidy syndrome identified by microcephaly on fetal ultrasound and confirmed by cytogenetic analysis of amniotic fluid. Initial mutational analysis of the entire coding region of the BUB1B gene failed to identify any causative mutations. However, further analysis revealed a known compound heterozygous mutation in the upstream region of this gene and a novel Alu insertion mutation in the intron.

Optical coherence tomography angiography and fundus autofluorescence in the eyes with choroideremia.

A 65-year-old man with presumed choroideremia with preserved central vision was examined by fundus autofluorescence (FAF) and optical coherence tomography angiography (OCTA). FAF showed an isolated area of hyperautofluorescence that involved the fovea. Although the choroid capillary slab of the OCTA showed the medium and large choroidal vessels inferior to the area of retinal pigment epithelium (RPE) atrophy, the choriocapillaris was visible in a relatively wider area than the hyperautofluorescent area in the FAF images. FAF and OCTA images allowed us to detect damage of the RPE before the choriocapillaris atrophy in a case of presumed choroideremia with preserved central vision.

Artificially reared mice exhibit anxiety-like behavior in adulthood.

It is important to establish experimental animal techniques that are applicable to the newborn and infant phases for nutrition and pharmacological studies. Breeding technology using the artificial suckling method without breast milk is very effective for the study of newborn nutrition. Using this method, we separated newborn mice from dams within 48 h of birth and provided them with artificial milk. We evaluated mouse anxiety levels after early postnatal maternal separation. Artificially reared mice were subjected to elevated plus-maze tests to assess emotional behavior at 9 weeks of age. Artificially reared mice showed a significantly lower frequency of entries and dipping into the open arms of the maze compared with dam-reared mice. This result indicates that the anxiety level of artificially reared mice was higher than that of dam-reared mice. Moreover, the concentration of monoamines in the brain was determined after the behavioral experiment. The hippocampal norepinephrine, serotonin, and 5-hydroxyindoleacetic acid levels in the artificially reared mice were significantly higher than those of the dam-reared mice. These results suggest that maternal-offspring interactions are extremely important for the emotional development of newborn infants during the lactation period. In future studies, it is necessary to consider the environmental factors and conditions that minimize the influence of artificial rearing on emotional behavior.

Experimental Verification of the Effects on Normal Domestic Cats by Feeding Prescription Diet for Decreasing Stress.

The objective of this study was to evaluate the effects of diet on the feline stress response by measuring plasma and urinary cortisol. A study diet was developed with a unique combination of nutrients that supports the management of stressful situations. The specific formulation of the diet included alpha-casozepine, which is believed to have an anxiolytic effect, and tryptophan supplementation. Tryptophan is the precursor for the synthesis of the neurotransmitter serotonin. Twenty-one indoor cats were fed with the study diet (n = 10) or a control diet (n = 11) for 8 weeks, after which physiological responses were evaluated. The study diet significantly increased the ratio of plasma tryptophan to large neutral amino acids and decreased urinary cortisol concentrations after being consumed daily for 8 weeks, but there was no effect on plasma cortisol levels following a stressful event (veterinary examination and blood draw). Further studies, such as behavioral analyses, are needed to clarify the effects of the study diet.

Simplification and transformation of ASTM F1292 measurement procedure for fall accident injury criteria.

Protecting children from injuries caused by fall accidents from playground equipment is important. Therefore, measures toward minimizing the risk of fall accident injuries are required. The risk of injury can be evaluated using ASTM F1292. In this test, G-max and the HIC are used to estimate the risk of injury. However, the measurement procedure is too complicated for application to a large number of installed equipment. F1292 requires simplified by reducing the number of phases, even with a small risk of loss in accuracy. With this in mind, this study proposes a shortened measurement procedure and a transformation equation to estimate the risk as same as F1292. As the result of experiments, it was revealed that G-max and the HIC values for both procedures linearly increase with drop height. The differences in outcomes between the regression equations of the standardized procedure and those of the shortened procedure can be used as a correction value. They can be added to the value measured by the shortened procedure. This suggests that the combination of the shortened procedure and transformation equation would be equivalent to F1292, with the advantage of being more easily and efficiently applied to the evaluation of installed playground equipment.

Central venous stenosis among hemodialysis patients is often not associated with previous central venous catheters.

It is widely assumed that central venous stenosis (CVS) is most commonly associated with previous central venous catheterization among the chronic hemodialysis (HD) patients. We evaluated the validity of this assumption in this retrospective study. The clinical records from 2,856 consecutive HD patients with vascular access failure during a 5-year period were reviewed, and a total of 26 patients with symptomatic CVS were identified. Combined with radiological findings, their clinical characteristics were examined. Only seven patients had a history of internal jugular dialysis catheterization. Diagnostic multidetector row computed tomography angiography showed that 7 of the 19 patients with no history of catheterization had left innominate vein stenosis due to extrinsic compression between the sternum and arch vessels. These patients had a shorter period from the time of creation of the vascular access to the initial referral (9.2 ± 7.6 months) than the rest of the patients (35.5 ± 18.6 months, p = 0.0017). Our findings suggest that cases without a history of central venous catheterization may not be rare among the HD patients with symptomatic CVS. However, those still need to be confirm by larger prospective studies of overall chronic HD patients with symptomatic CVS.

Reduced hydration status characterized by disproportionate elevation of blood urea nitrogen to serum creatinine among the patients with cerebral infarction.

The significance of fluid metabolism among the patients with cerebral infarction has barely mentioned in the literature despite the several reports suggesting the potential risk of reduced hydration status for the development of cerebral infarction. The aim of the this study is to explore the validity of the presumable relationship between hydration status and cerebral infarction. Ninety-seven patients with cerebral infarction from April 1, 2008 to March 31, 2009 were retrospectively investigated, and their hydration status were evaluated by using several clinical parameters such as a blood urea nitrogen to serum creatinine (BUN/Cr) ratio of >25 and plasma osmolality. Subjects with active infection, congestive heart failure, hepatic failure, gastrointestinal bleeding, or a malignancy were excluded since these conditions should modulate the absolute value of BUN/Cr ratio without a change in hydration status. Twenty-eight patients (29%) were considered as having reduced hydration status. The BUN/Cr ratio decreased significantly after the initiation of medical support (median 21.3; IR: 18.1-24.6), including oral or parenteral fluid supplementation, in comparison to the values at the time of patient admission (median 30.0; IR: 26.8-40.7; p<0.0001). Similar decreases were also observed in the hematocrit, hemoglobin, and plasma osmolality. The group considered to have reduced hydration status had a significantly higher prevalence of cardioembolic stroke than the other subjects. The hydration status may be a contributing factor to subtypes of cerebral infarction. Whether our findings are also the case with overall patients with cerebral infarction should be evaluated in greater detail.

[Case of lupus nephritis and enteritis associated with bilateral hydronephrosis].

A case of nephrotic syndrome associated with bilateral hydronephrosis in a 26-year-old female is reported. She was referred to our hospital because of persistent diarrhea, abdominal pain, and urinary disorders. On admission, ascites, intestinal edema, and bilateral hydronephrosis, were demonstrated by radiographic analysis. The findings of both physical and laboratory examinations showed evidence of systemic lupus erythematosus (SLE). In addition, diffuse proliferative lupus nephritis was consistently confirmed by a renal biopsy. Immediately after the initiation of steroid treatment, her abdominal symptoms disappeared followed by an improvement in the symptoms of intestinal edema, hydronephrosis, and the renal function. The relationship between ureterohydronephrosis and lupus cystitis, and the fact that lupus enteritis is often associated with lupus cystitis have been demonstrated by previous studies. Finally, the clinical manifestations observed in our case led us to consider the association of lupus enteritis and cystitis. We should bear in mind the possible association of several disorders, including nephrotic syndrome, enteritis, and hydronephrosis due to cystitis, in cases presenting with SLE.

[Case of Henoch-Schönlein purpura nephritis complicated with essential mixed cryoglobulinemia].

We report a case of purpura nephritis complicated with essential mixed cryoglobulinemia. The patient was referred to our hospital because of a petechial rash on the lower extremities, microscopic hematuria, and progressive deterioration of renal function. The presumptive diagnosis of Henoch-Schönlein purpura (HSP) was made, and the patient was treated with prednisolone at the dose of 40 mg/day. However, there was a persistent purpuric skin rash. On the other hand, immunoelectrophoresis of the serum revealed the presence of IgA-lambda and polyclonal IgG in the cryoprecipitate. Granular staining for polyclonal rather than monoclonal IgA and C3 segmentally along the capillary walls demonstrated by immunofluorescence analysis of renal biopsy led to the diagnosis of purpura nephritis as the major mechanism of renal damage. After three sessions of cryofiltration, the patient's serum cryoglobulins decreased and the active rash finally settled, along with improvement of renal function. These observations suggest that the presence of cryoglobulinemia modulated the clinical course of HSP in our case. Therefore, the possibility of the latent presence of cryoglobulinemia in cases with HSP having an active rash refractory to steroid treatment should not be overlooked.