iPSC-derived cortical neurons to study sporadic Alzheimer disease: A transcriptome comparison with post-mortem brain samples.

Alzheimer's disease (AD) is the most common cause of dementia, characterized by the progressive impairment of cognition and memory loss. Sporadic AD (sAD) represents approximately 95 % of the AD cases and is induced by a complex interplay between genetic and environmental factors called "Alzheimerogens". Heavy metals (e.g. copper) and pesticides (e.g. fipronil) can affect many AD-related processes, including neuroinflammation (considered as AD-inducing factor). Research would benefit from in vitro models to investigate effects of Alzheimerogens. We compared transcriptomics changes in sAD induced pluripotent stem cell (iPSC) derived cortical neurons to differentially expressed genes (DEGs) identified in post-mortem AD brain tissue. These analyses showed that many AD-related processes could be identified in the sAD iPSC-derived neurons, and furthermore, could even identify more DEGs functioning in these processes than post-mortem AD-brain tissue. Thereafter, we exposed the iPSCs to AD-inducing factors (copper(II)chloride, fipronil sulfone and an inflammatory cytokine cocktail). Cytokine exposure induced expression of immune related genes while copper-exposure affected genes involved in lipid and cholesterol metabolism, which are known AD-related processes. Fipronil-exposure did not result in significant transcriptomic changes, although prolonged exposures or higher doses may be necessary. Overall, we show that iPSC-derived cortical neurons can be beneficial in vitro models to identify Alzheimerogens and AD-related molecular mechanisms.

Molecular heterogeneity of second and fourth components of complement and their genes in systemic sclerosis and association of HLA alleles A1, B8 and DR3 with limited and DR5 with diffuse systemic sclerosis.

Deficiency of the complement component C4 at the functional, protein and gene level and deficiency of complement component C2 at the functional level were investigated and HLA analysis was performed on patients with limited and diffuse systemic sclerosis (SSc). One of the patients with limited SSc (n = 15) had subnormal C4, 1 subnormal C2 and 1 subnormal C4 and C2 activities; the latter patient had HLA alleles A11;B35;Dw1 associated with type II C2 deficiency and therefore most likely had a defect at the C2 locus. One of the patients with diffuse SSC (n = 12) had subnormal C4 and 1 subnormal C4 and C2 activities. C2 deficiencies in patients other than the one with the haplotype associated with C2 deficiency appeared not to be determined by the gene at the C2 locus. The incidence of partial C2 deficiency in a normal Caucasian population is reported to be 16 in 10,000, and that of partial C4 deficiency also appears to be very low. The percentages of C4A*Q0 and C4B*Q0 alleles in normal controls (n = 45) were within the reported range. Seven patients with limited SSc (n = 14) had one or two C4A*Q0 alleles and 2 with diffuse SSc (n = 13) had one C4A*Q0 allele. Thus, the incidence of C4A*Q0 was higher than normal in limited SSc and within the normal range in diffuse SSc. The two-sided Fisher's exact test applied on these data revealed that the association of C4A*Q0 with limited SSc did not reach a significant level (p = 0.10). Two of the 3 patients with limited SSc, who had two C4A*Q0 alleles, carried a heterozygous C4A-21-hydroxylase A (OHA) gene segment deletion as detected by Southern blotting. There was no correlation between the subnormal activity of C4 and the occurrence of one or two C4A*Q0 (and C4A-21-OHA segment deletion). HLA alleles A1, B8 and DR3 (p = 0.002) were associated with limited SSc (n = 23) and DR5(w11) (p = 0.018) with diffuse SSc (n = 17).

On the many faces of Leber hereditary optic neuropathy.

Leber hereditary optic neuropathy (LHON) is a maternally inherited disorder, associated with mutations in the mitochondrial DNA, which is notorious for its aspecific presentations. Two pedigrees are described with cases that are atypical for LHON with respect to sex, age of onset, interval between the eyes becoming affected, course of the disease, concomitant disorders, additional test results, final visual acuity, and/or results of mtDNA analysis. Moreover, the pedigrees themselves did not suggest maternal inheritance. We analysed the diagnostic and clinical genetic difficulties related to the atypical aspects of these pedigrees. We conclude that mtDNA analysis is justified in every case of optic nerve atrophy with no clear cause. Identification of one of the three LHON specifically associated mtDNA mutations is essential to confirm the diagnosis.

Incomplete functional deficiencies of the fourth (C4) and second (C2) components of complement in a patient with linear frontoparietal scleroderma and his family. Deficiencies determined by a gene not linked to human leukocyte antigen system.

BACKGROUND: In a previous study, a patient suffering from linear frontoparietal scleroderma and some of his family members were found to have an incomplete functional deficiency of the second component (C2) of complement (C). In this study, the proband and the rest of his family members were investigated for functional deficiencies of C2 and the fourth component of C (C4). A search for null alleles of C2 (C2*Q0) and C4 (C4*Q0) was made to find out whether their occurrence is responsible for incomplete functional deficiencies. HLA analysis was performed to find out whether deficiencies are linked to HLA alleles known to be associated with C4*Q0 and C2*Q0. Possible large deletions at C4 and 21-hydroxylase (21-OH) gene loci were also investigated in some family members. OBSERVATIONS: The proband had a combined functional deficiency of C4 and C2. Some of his family members had a partial functional deficiency of C4, some of C2 and some of C4 and C2; none had null alleles of C2 (C2*Q0), factor B (B*Q0) or C4B (C4B*Q0). C4*Q0 or functional C4 deficiency in this family was not associated with HLA-A1;B8;DR3 alleles. C2 deficiency was also not associated with HLA antigens known to be associated with type I and II C2 deficiencies. No gene deletion or unusual polymorphism at C4A and 21-OHA loci could be seen by restriction fragment length polymorphism (RFLP) studies. CONCLUSIONS: Combined and isolated partial functional deficiencies of C4 and C2 observed in the proband and many of his family members were not caused by C activation or null alleles. They were not linked to HLA system and were reminiscent of those observed previously in a family in which C4 deficiency was determined by a gene not linked to the HLA system.

Leber's hereditary optic neuropathy: no significant evidence for primary or secondary pathogenicity of the 15257 mutation.

Leber's hereditary optic neuropathy (LHON) is a maternally inherited disease of the optic nerves associated with various mitochondrial DNA (mtDNA) mutations. Four of these mutations, at nucleotide positions (np) 3460, 11778, 14484 and 15257, have been postulated to be of primary pathogenetical importance. Previously, we described the molecular and clinical findings in patients with the 11778 and 14484 mutations. Here we describe the molecular and clinical findings of patients in eight pedigrees with the 3460 mutation and in three pedigrees with the 15 257 mutation. In all three 15257 positive pedigrees the 3460, the 11778 or the 14484 mutation was also found. The first combination has not been reported before. We compared the clinical findings in these pedigrees with those of the 3460, 11778 and 14484 positive pedigrees that lack the 15257 mutation. No significant differences were found with respect to the age of onset, visual outcome or the probability of developing LHON. We conclude that there is no evidence that the 15257 mutation, which has been reported in normal controls, has primary causal significance, because it may coincide with the 3460, 11778 and 14484 mutations. We presume that the 15257 mutation has no secondary pathogenic importance, since it has no clear contribution to the degree or the probability of phenotypic expression.

[Enterobacterial sensitivity to colicins in the presence of enterochelin]. / Chuvstvitel'nost' énterobakterii k kolitsinam v prisutstvii énterokhelina.

The correlation between the colicine resistance of the reference and isolated strains of enterobacteria and their capacity for the biosynthesis of enterocheline, as well as the influence of exogenous enterocheline on the colicine sensitivity of enterobacteria were studied. In the wild strains of enterobacteria sensitivity to colicines was shown to have no correlation with capacity for the accumulation of catechol-type sideriphores. In some cases exogenous enterocheline prevents the lethal effect of colicines on the cultures of microorganisms capable of the biosynthesis of enterocheline.

[Formation of iron-binding metabolites by bacteria of the Enterobacteriaceae family]. / Obrazovanie zhelezosviazyvaiushchikh metabolitov bakteriiami semeistva Enterobacteriaceae.

The capacity of Enterobacteriaceae strains cultivated in a synthetic medium for the biosynthesis of catechols and hydroxamates has been studied. Among the strains under study all strains of the genera Salmonella (8 strains), Escherichia (102 strains), Citrobacter (5 strains), Enterobacter (2 strains), Serratia (1 strain) synthesize catechols. In the genus Shigella (128 strains) all Sh. flexneri serovars (79 strains) do not synthesize catechols, other representatives of this genus synthesize iron-fixing metabolites The synthesis of catecholsin the pathogenic Escherichia serovars is 1.5-2 times lower than in the non-pathogenic and opportunistic enterobacterial strains. Among the representatives of the genus Klebsiella (82 strains) catechols are synthesized by Kl. pneumoniae (68 strains) and not synthesized by kl. rhinoscleromatis (5 strains) and Kl. ozaenae (9 strains). Catechols are not synthesized also by all Proteus organisms under study (12 strains). All the enterobacteria under study show no capacity for the accumulation of hydroxamates under the conditions of catechol synthesis. The intensity of the synthesis of catechols depends on the composition of the medium and the conditions of cultivation.

The clinical value of the radioassay of serum folate.

Measurement of serum folate levels in humans using a radioassay has now been frequenly reported. However, very little data is available assessing the diagnostic value of this test in comparison with other laboratory and clinical information pertaining to folate status. A radioassay, using the folate binder in dried milk was used in parallel with the well established Lactobacillus casei microbiological assay. Three slightly different modifications of the radioassay were tested. At first, a partially purified milk binder was used, in the second the unpurified milk and, third, the unpurified milk with variation of the incubation periods during the assay, were utilized. A total of 159 sera were tested by one of the modifications of the radioassay as well as by the L. casei method. Duplicate estimations were done in all cases. Using the first method, the correlation between the radio- and microbiological assays was 0.89, with the second 0.52, and with the third 0.85. The final assay system gave the most reliable results when compared to the L. casei assay which was taken as the standard of reference. However, in a number of cases, normal levels by L. casei gave low levels by radioassay and vice versa. In 13 such instances, clinical and other laboratory data were assessed and it was concluded that the microbiological assay gave the more accurate indication of folate stores in humans. Some reasons for the discrepancies between the assay systems were discussed.