Induced sputum supernatant bioactive lipid mediators can identify subtypes of asthma.

BACKGROUND: Induced sputum (IS) allows to measure mediators of asthmatic inflammation in bronchial secretions. The specific role of induced sputum supernatant (ISS) endogenous bioactive lipid mediators in subtypes of asthma is not well understood. OBJECTIVE: To investigate the interactions between airway inflammation and clinical phenotypes of asthma, we integrated induced sputum supernatant (ISS) eicosanoids and quantitative assessment of infiltrating cells into new subtypes with the means of latent class analysis (LCA). METHODS: One hundred and thirty-nine asthmatics with and without aspirin hypersensitivity underwent sputum induction. High-performance liquid chromatography or gas chromatography coupled with mass spectrometry was used to profile eicosanoids. Nineteen variables covering clinical characteristics, IS inflammatory cells and eicosanoids were considered in the LCA. RESULTS: Four phenotypic asthma classes were distinguished. Class 1 with mild-to-moderate asthma, chronic rhinosinusitis (CRS), high PGA2 in ISS and almost equal distribution of inflammation cell patterns. Class 3 subjects also had mild-to-moderate asthma but without upper airway symptoms. Induced sputum was often paucigranulocytic with low levels of lipid mediators. Classes 2 and 4 represented severe asthma with CRS and impaired lung function despite high doses of steroids. High blood and sputum eosinophilia was in line with high cysteinyl leukotrienes and PGD2 in ISS only in class 2. Class 4 subjects tended to have increased sputum neutrophilia and PGE2 in ISS. Aspirin hypersensitivity was most frequent among class 2 subjects. CONCLUSIONS & CLINICAL RELEVANCE: The LCA revealed four distinct asthma classes differing in eicosanoid pathways.

Intermittent hypoxia regulates RNA polymerase II in hippocampus and prefrontal cortex.

Intermittent hypoxia (IH) is a major pathological factor in the development of neural deficits associated with sleep-disordered breathing. Here we demonstrate that IH lasting 2 or 30 days, but not sustained hypoxia (SH) of the same duration, was accompanied by several posttranslational modifications of the large subunit of RNA polymerase II, Rpb1, including hydroxylation of proline 1465, phosphorylation of serine 5 residues within the C-terminal domain, and nondegradative ubiquitylation. These modifications were found to occur in two regions of the brain, hippocampal region CA1 and the prefrontal cortex, but not in neocortex, brainstem and CA3 region of hippocampus. We also found that mice exposed to 14 or 30 days of IH, but not SH, demonstrated cognitive deficits in behavioral assays. Furthermore, by using the pheochromocytoma-derived PC12 cell line, we showed that, under in vitro IH conditions, induction of Rpb1 hydroxylation, phosphorylation, and ubiquitylation required that the von Hippel-Lindau protein be present. We hypothesize that the observed modifications of Rpb1 participate in regulating the expression of genes involved in mediating cognitive deficits evoked by chronic IH.

[Genetically determined familial male limited precocious puberty]. / Genetycznie uwarunkowane rodzinne przedwczesne dojrzewanie plciowe meskie.

Familial male-limited precocious puberty (FMPP) is a genetically determined, gonadotropin-independent disorder manifested by the sexual maturation at an early childhood. Low levels of blood gonadotropins, accompanied by a high testosterone levels, may suggest stimulation of androgen synthesis. The reports on the association of a constitutive activation of protein G-coupled receptors and pathogenesis of some clinical conditions have prompted searches for an abnormal structure of the LH/hCG receptor in the affected individuals. Point mutations, located mainly in the sixth transmembrane loop have been demonstrated to be responsible for the constitutive activation of the receptor and the cAMP-mediated stimulation of androgen formation in the Leydig cells.

Substitution of M398T in the second transmembrane helix of the LH receptor in a patient with familial male-limited precocious puberty.

Familial male-limited precocious puberty (MPP) is described in a 10 year old patient with typical symptoms of the disease. Sequence analysis of genomic DNA clearly demonstrated a heterozygous T1193C transition in exon 11 of the LH receptor (LHR) gene, which results in M398T substitution in the second transmembrane helix of the protein product of this gene. The same mutation was found in the patient's mother and in her brother. The grandmother and the relatives of the patient's father were free of the mutation. The boy was successfully treated with inhibitors of steroid biosynthesis and androgen antagonists. It is suggested that this mutation caused constitutive activation of the LHR, which results in excessive formation of androgens in Leydig cells and is responsible for the symptoms of precocious puberty in this patient. This is the second case of the familial form of MPP that was maternally inherited.

Study of the family of a patient with male-limited precocious puberty (MPP) due to T1193C transition in exon 11 of LH receptor gene.

Molecular diagnostics of the LHR gene was conducted in a 5-year-old boy with clinical symptoms and hormonal profile typical of precocious puberty. His parents and 4 sisters were also diagnosed. Single-strand conformation polymorphism analysis under temperature gradient conditions (Multitemperature SSCP) of 3 overlapping fragments of exon 11 of LHR gene revealed a mutation in the fragment spanning nucleotides 1072 to 1804. This mutation was found in the patient, in his mother and in his 4 sisters, and was confirmed by digestion with the use of restriction enzyme Bbr Cl. Direct sequencing revealed a heterozygous T1193C transition in the DNA fragment of the patient and in one of the alleles of his mother's and sister's DNA. This mutation causes Met398Thr substitution in the second transmembrane helix and results in a constitutive activation of LH receptor. This is the second identical mutation detected in Poland and one of the 7 identified so far in the world population.

Cloning and sequencing of the first plant GlnRS and GluRS genes.

We have cloned and sequenced glutamate-tRNA synthetase (GluRS) and glutaminyl-tRNA synthetase (GlnRS) from Arabidopsis thaliana. They have conservative motifs found in all known GlxRS genes. For Lupinus luteus we found only one gene of GlxRS. At the moment we do not know exactly, whether it corresponds to GlnRS or GluRS.

Distribution of 32 alelle of the CCR5 gene in the population of Poland.

The chemokine receptor CCR5 constitutes a major co-receptor for the R5 strains of HIV-1, and a mutant allele of the CCR5 gene, especially in the homozygous form delta32/delta32, confers resistance against infection by the virus. The frequency of the delta32 allele was determined in blood donors from 16 provinces, covering the entire territory of Poland. Among 861 individuals 182 (21.1%) were carriers of the mutated allele: 7 of them (0.8 %) were homozygotes delta32/delta32, and 175 (20.3%) were heterozygotes +/delta32, resulting in a 10.9% frequency of the delta32 allele. The highest frequencies of the mutated allele were found in the eastern and western provinces, and the lowest frequencies of the delta32 allele were detected in the provinces in the center of the country. This pattern of distribution may reflect the migration of the population from the eastern territories of Poland to the western part of the country after World War II.

A novel c.C2754 > T transition in the androgen receptor gene introduces the premature termination codon Q798X and results in a truncated form of the receptor.

A genetic analysis of a female with a 46,XY genotype and typical symptoms of the complete androgen insensitivity syndrome (CAIS) was conducted. The patient was diagnosed with an abdominal tumor due to the presence of a Sertoli cell adenoma in both gonads. Multiple temperature single-stranded conformational polymorphism (MSSCP) and sequence analyses of the androgen receptor gene revealed a c.C2754 > T mutation in exon 6. This mutation, which has not been previously reported, alters a Gln codon to a termination codon (Q798X). This results in the interruption of the amino acid sequence of the androgen receptor within the ligand-binding domain between helices VII and VIII. The truncated form of the receptor is devoid of 123 amino acids at the carboxyl end, a major part of the ligand-binding domain, and the AT2 sequence responsible for the activation of the transcription. It was concluded that the novel c.C2754 > T transition rendered the androgen receptor incapable of both ligand binding and activating the transcription, and was the cause of CAIS in the patient.