Knowledge and Compliance with Malaria National Treatment Guidelines among Primary Health Care Workers in a Rural Area in Northern Nigeria.

BACKGROUND: Knowledge and compliance with malaria treatment guidelines are among the major issues affecting treatment outcome for malaria in Nigeria. Primary health care (PHC) facilities are the first point of contact with the national health system for patients with malaria and other diseases. OBJECTIVE: This study assessed the knowledge and compliance with malaria National Treatment Guidelines (NTG) among PHC workers in Lere local government area of Kaduna State, North western Nigeria. METHODS: This was a descriptive cross-sectional study conducted among 42 community health workers. The total population of all eligible participants was used for subject selection. Data were analysed with SPSS IBM version 25.0 and STATA/SE 12. The level of statistical significance p-value was set at p<0.05. RESULTS: The mean age of the respondents was 38.02±9.23 years. Majority of the respondents were males (25; 59.5%) and community health extension workers (CHEWs) (24; 57.1%). Almost one-third (28.6%) of the PHC workers had poor knowledge of the recommendations of the NTG for malaria, while 14.3% had poor compliance with the NTG. Bivariate analysis showed a significant relationship between older age and good knowledge of the NTG ( χ2 =0.03, p=0.04). Multivariate analysis further revealed that the odds for poor knowledge of NTG was 40% higher among CHEWs compared to other health workers (AOR=1.40, 95% CI=0.25-7.93). The odds for good knowledge was lower by 55% among those who had practiced for <10 years compared to >10 years (OR=0.45, 95% CI=0.06-3.32). CONCLUSION: Poor knowledge and compliance to malaria NTG were commoner among lower cadre (CHEWs) staff with relatively fewer years in PHC practice. There is a need for training, retraining and equitable distribution of the NTG to ensure access and also improve knowledge and utilisation of the NTG for malaria by rural PHC workers.

CONTEXTE: La connaissance et le respect des directives de traitement du paludisme sont parmi les principaux problèmes qui affectent les résultats du traitement du paludisme au Nigéria. Les établissements de soins de santé primaires (SSP) sont le premier point de contact avec le système national de santé pour les patients atteints de paludisme et d'autres maladies. OBJECTIF DE L'ÉTUDE: Cette étude a évalué les connaissances et le respect des directives nationales de traitement du paludisme (NTG) parmi les travailleurs des SSP dans la zone de gouvernement local de Lere de l'État de Kaduna, dans le nord-ouest du Nigeria. MÉTHODES: Il s'agit d'une étude descriptive transversale menée auprès de 42 agents de santé communautaires. La population totale de tous les participants éligibles a été utilisée pour la sélection des sujets. Les données ont été analysées avec SPSS IBM version 25.0 et STATA/SE 12. Le niveau de signification statistique a été fixé à p<0,05. RÉSULTATS: L'âge moyen des personnes interrogées était de 38,02±9,23 ans. La majorité des personnes interrogées étaient des hommes (25 ; 59,5%) et des agents de vulgarisation en santé communautaire (24 ; 57,1%). Près d'un tiers (28,6 %) des agents de santé publique connaissaient mal les recommandations de la NTG pour le paludisme, tandis que 14,3 % ne respectaient pas la NTG. L'analyse bivariée a montré une relation significative entre l'âge avancé et la bonne connaissance du NTG ( χ2=0,03, p=0,04). L'analyse multivariée a également révélé que la probabilité d'une mauvaise connaissance de la NTG était 40% plus élevée chez les CHEW que chez les autres agents de santé (AOR=1,40, 95% CI=0,25-7,93). La probabilité d'une bonne connaissance était inférieure de 55% chez ceux qui avaient pratiqué pendant <10 ans par rapport à >10 ans (OR=0,45, 95% CI=0,06-3,32). CONCLUSION: Le manque de connaissances et d'observance des NTG sur le paludisme était plus fréquent parmi le personnel des cadres inférieurs (CHEWs) ayant relativement moins d'années de pratique dans les soins de santé primaires. Il est nécessaire de former, de recycler et de distribuer équitablement les NTG pour garantir l'accès et améliorer la connaissance et l'utilisation des NTG pour le paludisme par les travailleurs des SSP en milieu rural. Mots clés: Connaissance, Observance, Personnel de soins de santé primaires, Directives, Paludisme.

Relationship between the housing coldness/warmth evaluation by CASBEE Housing Health Checklist and psychological distress based on TMM Community-Based Cohort Study: a cross-sectional analysis.

OBJECTIVES: Previous studies have reported the relationship between housing environment and health, although due to cost and effort, it was difficult to conduct housing condition surveys on a large scale. The CASBEE Housing Health Checklist (the Checklist) made it possible to easily evaluate the housing condition from the resident's perspective. This study examined the relationship between housing coldness/warmth evaluation using the Checklist and psychological distress in a large-scale general Japanese population. STUDY DESIGN: A cross-sectional study. METHODS: We analysed data from 29,380 people aged ≥20 years who lived in Miyagi Prefecture, Japan. As an assessment of housing coldness/warmth, we used the Checklist. We classified participants' total scores on the Checklist related to coldness/warmth into quartiles. The Kessler 6 scale was used as an indicator of psychological distress. Multivariable logistic regression models were used to estimate the adjusted odds ratio (OR) and 95% confidence intervals (CIs). Adjusted OR and P-values for linear trends were calculated using the quartiles of the Checklists' score. RESULTS: Among participants in Q1 (i.e., poorer subjective house condition), the percentage of people with psychological distress was high. Compared to the highest quartile, Q1 showed poorer evaluation of housing coldness/warmth, and higher OR for psychological distress. The OR (95% CI) of psychological distress for Q3, Q2, and Q1 compared with Q4 were 1.93 (1.74-2.14), 2.82 (2.55-3.12), and 5.78 (5.25-6.35), respectively. CONCLUSIONS: Housing coldness/warmth evaluation was significantly related to psychological distress. This finding suggests that maintaining a comfortable thermal environment at home could be important for residents' mental health.

Earlier BMI rebound and lower pre-rebound BMI as risk of obesity among Japanese preschool children.

OBJECTIVES: Longitudinal growth data of children were analyzed to clarify the relationship between the timing of body mass index (BMI) rebound and obesity risk in later ages. SUBJECTS/METHODS: Of 54 558 children born between April 2004 and March 2005 and longitudinally measured in April and October every year in the preschool period, 15 255 children were analyzed wherein no longitudinal measurement is missing after 1 year of age. BMI rebound age was determined as the age with smallest BMI value across longitudinal individual data after 1 year of age. Rebound age was compared between overweight and non-overweight groups. The subjects were divided into groups based on the timing of rebound. The sex- and age-adjusted mean of the BMI, height and weight s.d. scores for age group, along with 6 months weight and height gain, were compared among groups using analysis of covariance. RESULTS: Among those who were overweight at 66-71 months of age, BMI rebound age obtained at approximately 3 years of age was compared with the non-overweight group, whose BMI rebound age was utmost 66 months or later (P<0.001). The comparison among BMI age group showed that earlier BMI rebound results in larger BMI (P<0.001) and larger weight and height gain after the rebound (P<0.001). Among the group with BMI rebound earlier than 30 months of age, low BMI was observed (P<0.001). Slight elevation of height and weight gain was observed before the BMI rebound among groups with rebound age earlier than 60 months of age (P<0.001). CONCLUSION: Earlier BMI rebound timing with pre-rebound low BMI leads to greater childhood obesity risk; hence, early detection and prevention is necessary for such cases.

Novel biallelic mutations in the PNPT1 gene encoding a mitochondrial-RNA-import protein PNPase cause delayed myelination.

Recent studies suggest that impaired transcription or mitochondrial translation of small RNAs can cause abnormal myelination. A polynucleotide phosphorylase (PNPase) encoded by PNPT1 facilitates the import of small RNAs into mitochondria. PNPT1 mutations have been reported in patients with neurodevelopmental diseases with mitochondrial dysfunction. We report here 2 siblings with PNPT1 mutations who presented delayed myelination as well as mitochondrial dysfunction. We identified compound heterozygous mutations (c.227G>A; p.Gly76Asp and c.574C>T; p.Arg192*) in PNPT1 by quartet whole-exome sequencing. Analyses of skin fibroblasts from the patient showed that PNPase expression was markedly decreased and that import of the small RNA RNaseP into mitochondria was impaired. Exogenous expression of wild-type PNPT1, but not mutants, rescued ATP production in patient skin fibroblasts, suggesting the pathogenicity of the identified mutations. Our cases expand the phenotypic spectrum of PNPT1 mutations that can cause delayed myelination.

Longitudinal changes in body mass index of children affected by the Great East Japan Earthquake.

BACKGROUND: The evacuation and disruption in housing caused by the 2011 Great East Japan Earthquake and following nuclear radiation may have influenced child health in many respects. However, studies regarding longitudinal childhood growth are limited. Therefore, in this study we aimed to explore the influence of the earthquake on longitudinal changes in body mass index in preschool children. METHODS: Participants were children from nursery schools who cooperated with the study in the Iwate, Miyagi and Fukushima prefectures. The exposed group consisted of children who experienced the earthquake during their preschool-age period (4-5 years old). The historical control group included children who were born 2 years earlier than the exposed children in the same prefectures. Trajectories regarding body mass index and prevalence of overweight/obesity were compared between the two groups using multilevel analysis. Differences in the changes in BMI between before and after the earthquake, and proportion of overweight/obesity was compared between the two groups. We also conducted subgroup analysis by defining children with specific personal disaster experiences within the exposed group. RESULTS: A total of 9722 children were included in the study. Children in the exposed group had higher body mass indices and a higher proportion of overweight after the earthquake than the control group. These differences were more obvious when confined to exposed children with specific personal disaster experiences. CONCLUSIONS: Children's growth and development-related health issues such as increased BMI after natural disasters should evoke great attention.

Toll-like receptor 3 in nasal CD103+ dendritic cells is involved in immunoglobulin A production.

Intranasal inoculation with influenza hemagglutinin subunit with polyinosine-polycytidylic (polyI:C), a synthetic analog for double-stranded RNA, enhances production of vaccine-specific immunoglobulin (Ig) A, which is superior to IgG in prophylactic immunity. The mechanism whereby polyI:C skews to IgA production in the nasal-associated lymph tissue (NALT) was investigated in mouse models. Nasally instilled polyI:C was endocytosed into CD103+ dendritic cells (DCs) and induced T-cell activation, including interferon (IFN)-γ production. According to knockout mouse studies, polyI:C activated the Toll-like receptor 3 signal via the adapter TICAM-1 (also called TRIF), that mainly caused T-cell-dependent IgA production. Nasal CD103+ DCs activated transforming growth factor-ß signaling and activation-induced cytidine deaminase upon polyI:C stimulation. IgA rather than IgG production was impaired in Batf3-/- mice, where CD103+ DCs are defective. Genomic recombination occurred in IgA-producing cells in association with polyI:C-stimulated DCs and nasal microenvironment. PolyI:C induced B-cell-activating factor expression and weakly triggered T-cell-independent IgA production. PolyI:C simultaneously activated mitochondrial antiviral signaling and then type I IFN receptor pathways, which only minimally participated in IgA production. Taken together, CD103+ DCs in NALT are indispensable for the adjuvant activity of polyI:C in enhancing vaccine-specific IgA induction and protective immunity against influenza viruses.

Identification of a common mutation in Finnish patients with nonketotic hyperglycinemia.

Nonketotic hyperglycinemia (NKH) is an autosomal recessive metabolic disorder caused by the defects in the glycine cleavage system (GCS; EC 2.1.2.10), a multienzyme system that consists of four individual components. NKH is a rare disorder in many countries, but with a very high incidence in northern Finland. To understand the genetic background of this high incidence, we examined the GCS in a typical case of NKH at the molecular level. The activity of P protein, a component of the GCS, was not detected in the lymphoblasts of the patient, while P protein mRNA of a normal size and level was present in the cells. Structural analysis of P protein mRNA from the patient revealed a single nucleotide substitution from G to T in the protein coding region, which resulted in an amino acid alteration from Ser564 to Ile564. No P protein activity was detected when the mutant P protein with this amino acid substitution was expressed in COS 7 cells. The patient was homozygous for this mutation. Furthermore, this mutation was present in 70% (14 of 20) of P protein gene alleles in Finnish patients with NKH, whereas it was not found in 20 alleles of non-Finnish patients. The results suggest that this mutation is responsible for the high incidence of NKH in Finland.

Normal hepatic hemodynamics during early postoperative period in recipients with adult live donor liver transplantation.

To recognize "normal" hepatic hemodynamics after live donor liver transplantation (LDLT), we analyzed Doppler parameters on recipients with a right liver graft and donors after extended left hepatectomy. Theoretically these values should be the same. From April 2000 to October 2004, 20 LDLTs were performed using a right liver graft. The 10 recipients without postoperative complications and their donors were included in this study. Portal venous velocity (PVV; cm/s), hepatic arterial peak systolic velocity (cm/s), and hepatic venous peak velocity (HVPV; cm/s) were measured during the first 2 weeks. In donors PVV and HVPV after LDLT were significantly higher after than before left hepatectomy: 19.2 +/- 4.2 vs. 31.5 +/- 13.0 cm/s (P = .013) and 23.0 +/- 7.2 vs. 41.8 +/- 10.3 cm/s respectively (P = .010). However, there were mild degrees of increased PVV and HVPV. In recipients, a markedly increased PVV (106.3 +/- 45.2 cm/s on day 1) was significantly higher than that in donors on each postoperative day. The hepatic arterial resistive index in recipients was also significantly higher than that in donors on each postoperative day, for example, 0.72 +/- 0.11 vs 0.62 +/- 0.04 on day 1 (P = .0326). In conclusion, we have shown "abnormal" hepatic hemodynamics in even those recipients without complications during the early postoperative period after LDLT.

Association of a mutation in thiazide-sensitive Na-Cl cotransporter with familial Gitelman's syndrome.

Gitelman's syndrome is a variant of Bartter's syndrome, characterized by hypokalemia, hypomagnesemia, hypocalciuria, and hypovolemia. We have observed familial cases of Gitelman's syndrome, and a possible mutation in thiazide-sensitive Na-Cl cotransporter was investigated in this kindred. The proband was a 47-yr-old Japanese female, and her mother was also affected. Her parents and maternal grandparents are consanguineous. By using PCR-amplification and direct sequencing, we identified a novel non-conservative missense mutation at 623 amino acid position, which substitutes proline for leucine (L623P), and also creates an Nci I restriction site in the exon 15. The mutation was not detected in normal healthy subjects (n = 102). Nci I digestion of PCR-amplified exon 15 DNA fragments from individuals in the family indicated the autosomal recessive inheritance of the disorder. In conclusion, the L623P mutation in the thiazide-sensitive Na-Cl cotransporter gene is suggested to impair the transporter activity, and to underlie this familial Gitelman's syndrome; Gitelman's syndrome observed in this kindred has been inherited in an autosomal recessive fashion.

Inhibition of Epstein-Barr virus infection in vitro by recombinant human interferons alpha and gamma.

The inhibitory effects of pure recombinant human interferons alpha A and gamma (reIFN-alpha A and -gamma) on Epstein-Barr virus (EBV) infection of a human EBV-negative B cell line, BJAB, and of normal adult B lymphocytes were studied. With pretreatment for 24 h, both types of reIFNs were effective in suppressing the production of EBV specific nuclear antigen (EBNA-1) in BJAB cells 24 h after EBV-infection, as determined by the immunoblotting technique. ReIFN-alpha A was, however, a much more potent inhibitor than reIFN-gamma. With treatment starting 1 h after EBV infection, both types of reIFNs were less effective in the suppression of EBNA production. Neither of the reIFNs showed any inhibitory effect on EBNA production in the latently EBV-infected cell lines, Raji and Daudi. These results suggest that reIFNs act in the early phase of EBV infection. Both types of reIFNs were also effective in inhibiting EBV infection of normal adult B lymphocytes as demonstrated by a reduction both in [3H]thymidine incorporation 6 days after EBV infection and in the total number of proliferating cells 21 days after EBV infection. Again, reIFN-alpha A showed a greater inhibitory effect than reIFN-gamma. We also showed that in BJAB cells, reIFN-alpha A strongly induced (2'-5')oligoadenylate synthetase activity, whereas reIFN-gamma increased the surface expression of HLA class I antigens.

Novel mutations in the connexin 26 gene (GJB2) responsible for childhood deafness in the Japanese population.

Mutations in the connexin 26 gene (GJB2), which encodes a gap-junction protein and is expressed in the inner ear, have been shown to be responsible for a major part of nonsyndromic hereditary prelingual (early-childhood) deafness in Caucasians. We have sequenced the GJB2 gene in 39 Japanese patients with prelingual deafness (group 1), 39 Japanese patients with postlingual progressive sensorineural hearing loss (group 2), and 63 Japanese individuals with normal hearing (group 3). Three novel mutations were identified in group 1: a single nucleotide deletion (235delC), a 16-bp deletion (176-191 del (16)), and a nonsense mutation (Y136X) in five unrelated patients. The 235delC mutation was most frequently observed, accounting for seven alleles in 10 mutant alleles. Screening of 203 unrelated normal individuals for the three mutations indicated that the carrier frequency of the 235delC mutation was 2/203 in the Japanese population. No mutation was found in group-2 patients. We also identified two novel polymorphisms (E114G and I203T) as well as two previously reported polymorphisms (V27I andV37I). Genotyping with these four polymorphisms allowed normal Japanese alleles to be classified into seven haplotypes. All 235delC mutant alleles identified in four patients resided only on haplotype type 1. These findings indicate that GJB2 mutations are also responsible for prelingual deafness in Japan.

Glycogen storage disease type Ib: structural and mutational analysis of the microsomal glucose-6-phosphate transporter gene.

Glycogen storage disease type Ib is caused by a mutation in the gene encoding microsomal glucose-6-phosphate (G6P) transporter. We determined the exon/intron organization of the G6P transporter gene. Four overlapping genomic fragments containing the entire coding region of the gene were amplified by polymerase chain reaction (PCR) using exonic primers, and their nucleotide sequences were determined. The gene spans 4.5 kb and has eight exons. All exon/intron boundaries adhered to the canonical AG/GT rule. We then designed eight pairs of PCR primers to amplify all coding exons for a mutational analysis and studied five Japanese patients with the disease. Two novel homozygous mutations were identified in two families: a three-base deletion (delV235) in exon 2 in a consanguineous family and a splicing mutation (IVS7+1G-->T) in intron 7 in a nonconsanguineous family. Patient 3 was a compound heterozygote of W118R and IVS1+1G-->A, both of which we previously identified [Kure et al., 1998: Biochem Biophys Res Commun 248:426-431]. Patients 4 and 5 were homozygotes of W118R. Including our previous study, we found a total of ten W118R alleles in nine Japanese patients. The results support our previous suggestion that W118R is prevalent among Japanese patients. The genomic sequence data and mutation spectrum obtained from the Japanese patients will facilitate genetic diagnosis of glycogen storage disease type Ib.

Heterogeneous mutations in the glucose-6-phosphatase gene in Japanese patients with glycogen storage disease type Ia.

Glycogen storage disease type Ia (GSD-Ia) is an autosomal recessive disorder of glycogen metabolism caused by glucose-6-phosphatase (G6Pase) deficiency. It is characterized by short stature, hepatomegaly, hypoglycemia, hyperuricemia, and lactic acidemia. Various mutations have been reported in the G6Pase gene (G6PC). However, in Japanese patients, a g727t substitution was found to be the major cause of GSD-Ia, accounting for 20 of 22 mutant alleles [Kajihara et al., 1995], and no other mutations have been found in this population. We analyzed four Japanese GSD-Ia patients and identified three other mutations in addition to the g727t. They included two missense mutations (R83H and P257L) and one nonsense mutation (R170X). Each of the three mutations exhibited markedly decreased G6Pase activity when expressed in COS7 cells. A patient homozygous for R170X showed multiple episodes of profound hypoglycemia associated with convulsions, while P257L was associated with a mild clinical phenotype. The presence of R170X in three unrelated families may implicate that it is another important mutation in the etiology of GSD-Ia in Japanese patients. Thus, the detection of non-g727t mutations is also important in establishing the DNA-based diagnosis of GSD-Ia in this population.

Glycogen storage disease type Ia: molecular diagnosis of 51 Japanese patients and characterization of splicing mutations by analysis of ectopically transcribed mRNA from lymphoblastoid cells.

Glycogen storage disease type Ia (GSD-Ia) is an autosomal recessive disorder of glycogen metabolism caused by a deficiency of glucose-6-phosphatase (G6Pase) that is expressed in the liver, kidney, and intestinal mucosa. Clinical manifestations include short stature, hepatomegaly, hypoglycemia, hyperuricemia, and lactic acidemia. To elucidate a spectrum of the G6Pase gene mutations and their frequencies, we analyzed mutations in 51 unrelated Japanese patients with GSD-Ia. The most prevalent mutation was g727t, accounting for 88 of 102 mutant alleles examined, followed by R170X mutation, which accounted for 6 mutant alleles, and R83H mutation which was observed in 3 mutant alleles. In addition, 3 different, novel mutations, IVS1-1g

Structure and expression of the glycine cleavage system in rat central nervous system.

The glycine cleavage system (GCS) is a mitochondrial multienzyme system consisting of four individual proteins, three specific components (P-, T-, and H-proteins) and one house-keeping enzyme, dihydrolipoamide dehydrogenase. Inherited deficiency of the GCS causes nonketotic hyperglycinemia (NKH), an inborn error of glycine metabolism. NKH is characterized by massive accumulation of glycine in serum and cerebrospinal fluids and severe neuronal dysfunction in neonates. To elucidate the neuropathogenesis of NKH, we cloned cDNAs encoding three specific components of the GCS and studied the gene expression in rat central nervous system. P-, T-, and H-protein cDNAs encoded 1024, 403, and 170 amino acids, respectively. In situ hybridization analysis revealed that P-protein mRNA was expressed mainly in glial-like cells, including Bergmann glias in the cerebellum, while T- and H-protein mRNAs were detected in both glial-like cells and neurons. T- and H-protein mRNAs, but not P-protein mRNA, were expressed in the spinal cord. Primary astrocyte cultures established from cerebral cortex had higher GCS activities than hepatocytes whereas those from spinal cord expressed only H-protein mRNA and had no enzymatic activity. An important role of glycine as inhibitory neurotransmitter has been established in the brainstem and spinal cord and another role of glycine as an excitation modulator of N-methyl-D-aspartate receptor is suggested in the hippocampus, cerebral cortex, olfactory bulbus, and cerebellum. Our results suggest that the GCS plays a major role in the forebrain and cerebellum rather than in the spinal cord, and that N-methyl-D-aspartate receptor may participate in neuropathogenesis of NKH.

Endonuclease activation following focal ischemic injury in the rat brain.

The structural changes which occur in chromatin DNA after ischemic brain injury are poorly understood. This study examined the appearance of double-strand DNA breaks and the temporal profile of DNA degradation following focal ischemic injury in rat brain. Focal cerebral ischemia was produced by tandem occlusion of the common carotid and proximal middle cerebral arteries. The effects of decapitation ischemia were also studied by DNA analysis. DNA was extracted by standard methods from the ischemic brain tissues and electrophoresed on a 1.5% agarose gel. With decapitation ischemia, random DNA cleavage was observed as a dense "smear" on the gel electrophoresis beginning 6 h after the ischemic insult, and increasing in amount thereafter. Focal ischemia provided DNA fragmentation, which is specific DNA cleavage at the internucleosomal linker regions, particularly in the caudoputamen. Coexisting random degradation and specific fragmentation of DNA was observed in the cortex following focal ischemia. To determine whether an endonuclease responsible for DNA fragmentation was present, nuclear proteins were extracted from normal brain nuclei and the endonuclease activity was determined using plasmid DNA and a nuclear incubation system. This demonstrated that brain nuclear proteins have Ca(2+)-dependent endonuclease activity which is related to DNA fragmentation. Ischemic injury causes both random and specific DNA cleavage in the brain, which is probably mediated by Ca(2+)-dependent endonuclease.

Apoptotic cell death of a temperature-sensitive central neuronal cell line.

A neuron-like cell line HS-2, derived from a primary fetal rat (E17) hippocampal cell culture using the temperature-sensitive SV 40 large T antigen, exhibits flat shape and grows well in culture medium with 5% fetal calf serum (FCS) at the permissive temperature (PT, 33.5 degrees C). At the non-permissive temperature (NPT, 38.5 degree C), many, but not all cells, have a neuronal shape with processes. The addition of dibutyryl-cAMP promotes the morphological changes in the cells to a neuron-like shape with long neurite-like processes and the cells exhibit neuron-specific enolase- and glutamic acid decarboxylase-immunoreactivity. Apoptotic cell death also occurs in these cultures at the NPT. DNA fragmentation and chromatin condensation that are characteristic of apoptosis occur within 8 h of being placed at the NPT. By 48 h after being placed at the NPT, the number of surviving cells decreases by 40% in the presence of 5% FCS. This cell line should be useful for investigating the mechanisms of 'programmed cell death' of neurons, which appears to occur during brain development and possibly in CNS degenerative diseases.

DNA fragmentation in focal cortical freeze injury of rats.

This study examined the appearance of double-strand DNA breaks in rat brain after a focal cortical freeze injury in vivo. DNA fragments of oligonucleosome size appeared 3 h after the injury, and increased in a time-dependent manner. At 24 h, the amount of DNA fragmentation reached a maximum and then declined. When nuclei from freeze-injured brain tissue were incubated with Ca2+ in vitro, increased endonuclease activity, which can cause DNA fragmentation, was found. These findings indicate that the activation of a Ca(2+)-dependent endonuclease may be involved in the evolution of freeze-traumatized brain tissue.

A novel mutation in glial fibrillary acidic protein gene in a patient with Alexander disease.

Alexander disease is a rare, progressive, leukoencephalopathy whose hallmark is the widespread accumulation of Rosenthal fibers. The most common form affects infants and young children, and is characterized by progressive failure of central myelination, usually leading to death before adulthood. Definitive diagnosis of Alexander disease has required biopsy or autopsy to demonstrate the presence of Rosenthal fibers. However, missense mutations in the coding region of the glial fibrillary acidic protein (GFAP) gene have recently been associated with a high percentage of pathologically proven cases. Here we report that a 10-year-old Japanese patient who showed clinical signs of Alexander disease is heterozygous for a C to T transition in which predicts a novel A244V amino acid substitution in the conserved 2A alpha-helix domain of GFAP. The nucleotide change was not found in 65 normal individuals (130 alleles). These results provide further support for a causative role for GFAP mutations in Alexander disease, and suggest DNA sequencing as an alternative diagnostic to biopsy.