Introduction. Acute intermittent porphyria (AIP) is the most common and severe form of acute hepatic porphyria. AIP is caused by a deficiency in the third enzyme of the heme biosynthesis system — hydroxymethylbilanine synthase (HMBS) — and has a dominant inheritance type. However, the probability of the clinical manifestation of this condition in carriers of the mutation in the HMBS gene constitutes only 10–20 %. Thi s suggests that the presence of such a mutation can be a necessary but not a sufficient condition for the development of the disease.Aim. To search for additional genetic factors, which determine the clinical penetrance of AIP using Whole-Exome Sequencing.Materials and methods. Sequencing of the whole exome was performed using a TruSeqExomeLibraryPrepkit (Illumina) kit by an Illumina HiSeq4000 instrument for 6 women with API with known mutations in the HMBS gene. All the patients suffered from a severe form of the disease. As a reference, a version of the hg19 human genome was used.Results. No common mutations were found in the examined patients. However, in each patient, functional variations were found in the genes related to detoxification systems, regulation of the heme biosynthesis cascade and expression of delta-aminolevulinic acid synthase (ALAS1) and in genes of proteins regulating nervous system. These variations require further study involving an extended number of patients with AIP manifestations and their relatives, who are asymptomatic carriers of disorders in the gene HMBS.Conclusions. The results obtained have allowed us to formulate a hypothesis about a possible role of genetic defects in the penetrance of AIP, which determine the development of other neurological pathologies. This is evidenced by the presence of gene pathogenic variations in 5 out of 6 examined patients, defects in which are associated with hereditary myasthenia and muscle atrophy.

Acute intermittent porphyria (AIP) is caused by the partial deficiency of porphobilinogen deaminase (PBGD), one of the enzymes of the heme biosynthetic pathway. The penetrance of the mutant gene PBGD is not high and averages of 10-15%. Any additional genetic factors, the combination of which with the mutant allele of the PBGD gene leads to the clinical manifestation of AIP is not currently known. The eventual associations of allelic variants of genes of the Phase 1: CYP1A1 (A2455G), CYP2E1 (G1259C) and four genes of the phase 2: NAT2 (C481T, G590A G857A), mEPHX1:Tyr113His - 3rd exon, His139Arg - 4th exon, GSTM1 (Del), GSTT1 (Del) with clinical presentation of AIP were investigated. Homozygous carriership of the "fast" allele of the acetyltransferase gene (genotype N/N) was established to be associated with a latent course of the disease. The combination of "functionally weakened" genotypes of glutathione transferase (class t similar to) and class M (GSTT10/0, GSTM10/0) can be considered as an unfavorable genetic factor related with the clinical presentation of AIR Comparative analysis of the frequencies of genotypes and polymorphic alleles of genes CYP1A1, CYP2E1 and mEPHX1 revealed no statistically significant differences between the samples of patients with AIP and asymptomatic carriers of the disease.

Aim. To evaluate the impact of developing pregnancy on porphyrin metabolism in reproductive-aged women with acute porphyria (AP).Subjects and methods. The prospective clinical data of 33 pregnancies were analyzed in 28 patients with the established diagnosis of AP. The latter was verified by the quantitative analysis of 24-hour urinary porphyrin excretion and the diminished activity of the pathognomonic enzyme.Results. Each case was analyzed in detail according to different criteria. Poor prognostic factors for pregnancy are identified in AP. The used curation policy for pregnant patients is described. The pregnant women with occurring AP episodes are subdivided into clinical groups requiring different curation approaches. The scheme for the used working protocol is given.Conclusion. The accumulated experience with curating the patients with AP will be able to avoid the existing prohibitory practice, providing a way to develop a new quality of life in the patients' families.

Acute intermittent porphyria (AIP) is an autosomal dominant hereditary disease, caused by partial deficiency of porphobilinogen deaminase (PBGD), one of the key enzymes of heme biosynthesis. This study describes molecular genetics of AIP in Russia. Mutation analysis of PBGD gene in 70 unrelated patients revealed 47 various genetic defects, 28 of which had not been described previously. Mutations 53delT and Arg173Trp (recorded 8 times, in total 23%) proved to be the most common in Russia. Microdeletion 53delT has monophyletic origin and was found only in Russia. Molecular genetic examination of 132 relatives of AIP patients from 40 families revealed 52 latent carriers of the disease. Low (about 10%) AIP penetrance indicates that a mutation in the PBGD gene is an important but not sufficient prerequisite for clinical manifestation of the disease. Modulation of penetrance in erythropoietic protoporphyria by coinheritance of a mutant allele and a functionally defective wild type allele of ferrochetalase gene has been shown previously. We hypothesized that similar mechanism works in AIP. Sequencing of the full length PBGD genes from unrelated AIP patients as well as SNP analysis, and the analysis of abnormal PBGD mRNA splicing showed that in case of AIP, this hypothesis is not true and some other factors are responsible for the penetrance of this disease.

A case of acute intermittent porphyria is described in a 37-year-old female patient treated with normasang, a drug that regulates porphyrin metabolism at the last stages of the disease. Chronic renal failure with the hypertensive syndrome, severe neurological symptoms, and vascular sclerotic changes in all organs were the symptoms of the underlying disease. Infectious complications were the cause of sepsis and favoured deteriorated multiple organ dysfunction that determined lethal exitus.

It is recommended to follow up patients with acute intermittent porphyria (AIP) both in clinical improvement and during an acute attack. Knowledge of the specificity of porphyrin metabolism at different disease stages can help in adequate evaluation of the AIP course. Investigation of porphyrin metabolism in 43 patients with AIP at different stages of the disease was made. Porphyrin metabolism was evaluated by the levels of total porphyrins and their precursors (porphobilinogen and delta-aminolevulinic acid) in the urine, concentration of total porphyrins in the feces, plasm, red cells, activity of porphobilinogen deaminase in the red cells. It was established that metabolism of porphyrins in patients with acute AIP and a few years after it is characterized by high and unstable values. With time, in strict adherence to the hematologist's recommendations porphyrin metabolism stabilizes on higher levels and the risk of new attacks decreases. The algorithm of follow-up at different stages of the disease is proposed.

AIM:To characterize patients with various nosological unities [symbol: see text] of porphyria in accordance with their age, clinical symptoms, provoking factors, therapy and outcome.MATERIAL AND METHODS:Patients with acute intermittent porphyria (43), hereditary coproporphyria (8), variegate porphyria (3), porphyria cutanea tarda (7), hepatoerythropoietic porphyria (1), and hereditary erythropoietic porphyria (2) were studied. One patient was suspected of porphyria caused by deficiency of delta-aminolevulenic acid dehydrogenase.RESULTS:The patients were from the CIS. The overwhelming majority of them were young and middle-aged subjects. Rapid development of the disease and severe neurological symptoms were predominantly observed in patients with acute forms of porphyria.CONCLUSION:Early diagnosis of porphyrin metabolism disorders makes it possible to decrease abruptly the number of cases leading to severe complications, disability, and fatal outcome. The use of inexpensive methods of screening of porphyrin metabolism disorders provides a promising approach to solving this problem. These methods should be used in municipal hospitals. In addition, asymptomatic carriers of defective gene should be revealed at the preclinical stage using various methods of molecular genetic assay.

Porphobilinogen deaminase (PBGD) is a key enzyme of the heme biosynthetic pathway. Defects in the PBGD gene lead to an autosomal dominant disease, acute intermittent porphyria (AIP). Almost all AIP patients with rare exceptions are heterozygous for the defective gene. To date, at least 160 different mutations causing AIP are identified. Extensive investigations along this line are conducted in many countries of the world. In Russia these studies had not been hitherto performed. Here we report the results of molecular genetic examination of four Russian patients with AIP diagnosed from clinical symptoms. By direct sequencing of the PBGD gene or the corresponding cDNA, we have detected four mutations, three of which were not previously encountered in the world population. These are TAAG deletion in intron 7 between positions +2 and +5 (IVS7 2–5 delTAAG); T deletion in the initiation codon ATG of exon 3, and the G for C replacement at position –1 of intron 5 (IVS5 as –1 G–C), which disrupts splicing. In addition, in one female patient, a known deletion CT in codon 68 was revealed. In two patients, expression of PBGD gene alleles was significantly disproportional, so that normal mRNA prevailed in one case and mutant mRNA of nonerythroid type in the other. Deletion in intron 7 was easily detectable due to the formation of a heteroduplex fragment with abnormal electrophoretic mobility directly in PCR. This simple heteroduplex analysis allowed us to exclude AIP carriage in son and daughter of a female patient with the genetic defect.

Porphobilinogen deaminase (PBGD) is a key enzyme of the heme biosynthetic pathway. Defects in the PBGD gene lead to an autosomal dominant disease, acute intermittent porphyria (AIP). Almost all AIP patients with rare exceptions are heterozygous for the defective gene. To date, at least 160 different mutations causing AIP are identified. Extensive investigations along this line are conducted in many countries of the world. In Russia these studies had not been hitherto performed. Here we report the results of molecular genetic examination of four Russian patients with AIP diagnosed from clinical symptoms. By direct sequencing of the PBGD gene or the corresponding cDNA, we have detected four mutations, three of which were not previously encountered in the world population. These are TAAG deletion in intron 7 between positions +2 and (IVS7 2-5 delTAAG); T deletion in the initiation codon ATG of exon 3, and the G for C replacement at position -1 of intron 5 (IVS5 as -1 G:C), which disrupts splicing. In addition, in one female patient, a known deletion CT in codon 68 was revealed. In two patients, expression of PBGD gene alleles was significantly disproportional, so that normal mRNA prevailed in one case and mRNA of nonerythroid type in the other. Deletion in intron 7 was easily detectable due to the formation of a heteroduplex fragment with abnormal electrophoretic mobility directly in PCR. This simple heteroduplex analysis allowed us to exclude AIP carriage in son and daughter of a female patient with the genetic defect.

A comparative analysis was made of peptide components of low-molecular fractions of red cell lysate from 7 donors and 18 cancer patients (7 patients with lymphosarcoma, 5 patients with newly diagnosed cancer of the lungs and 6 patients with newly diagnosed cancer of the stomach). Lymphosarcoma and lung carcinoma patients are shown to have defects of intraerythrocytic proteolysis of hemoglobin leading to a rise in the amount of some peptide components. Each type of the disease is characterized by particular alterations in peptide composition of the red cells. These changes were absent in patients with gastric cancer.

Clinicohematological investigations and cytogenetic analysis of blood lymphocytes were made 5-7 years after the Chernobyl accident in 201 liquidators who had worked in the radionuclide-contaminated zone. Among the somatic diseases found in the examinees statistically more prevalent were cardiovascular and gastrointestinal affections, asthenic syndrome, thyroid disorders. Hemograms presented a rise in hemoglobin, red cell and eosinophil content, a drop in the number of neutrophils. A tendency to erythrocytosis was observed in 20.3% of the wreckers. Dicenters and rings were abundant in the lymphocytes of 69% of the cytogenetically examined examinees 5-7 years after the exposure to radiation.

Clinicohematological investigations and cytogenetic analysis of blood lymphocytes were made 5-7 years after the Chernobyl accident in 201 liquidators who had worked in the radionuclide-contaminated zone. Among the somatic diseases found in the examinees statistically more prevalent were cardiovascular and gastrointestinal affections, asthenic syndrome, thyroid disorders. Hemograms presented a rise in hemoglobin, red cell and eosinophil content, a drop in the number of neutrophils. A tendency to erythrocytosis was observed in 20.3% of the wreckers. Dicenters and rings were abundant in the lymphocytes of 69% of the cylogenetically examined examinees 5-7 years after the exposure to radiation.

To evaluate oxygen transport function of Hb in lymphoid sarcomas, 15 patients aged 16-63 were examined for Hb oxygen capacity, concentrations of fetal Hb and creatinin, basic ligands H+, CO2, 2,3-diphosphoglycerate (2,3-DPG). Oxy-Hb dissociation curves were plotted. The patients' blood contained HbF in elevated quantities, whereas red cells had high creatinin. This emphasizes the role of hemolysis in genesis of anemia in lymphosarcomas. 70% of the examinees retained normal Hb affinity to oxygen in vivo, in 30% this affinity exceeded the standard. Compensatory mechanism responsible for low Hb affinity to oxygen in this disease fails. Therefore lymphosarcoma patients with anemia need early replacement hemotransfusions.

To define Hb role in compensation of hemic hypoxia, the authors studied HbO2 dissociation curves (HDC), blood acid-base equilibrium, concentrations of 2,3-DPG and creatinine in red cells from females with moderate iron-deficiency anemia resultant from frequent haemorrhages. This form of anemia was found associated with a 34% increase in the level of 2,3-DPG against normal amounts of other two ligands essential to Hb affinity to oxygen, i.e. H+ ions and CO2. A 3-fold increase in creatinine concentrations and its direct relationship with 2,3-DPG quantitative characteristics gave ground for considering rejuvenescence of the red cells in the peripheral blood to be the underlying factor in enhancement of 2,3-DPG synthesis. A 2,3-DPG-induced in vivo HDC shift to the right by 2.5 mmHg stimulates oxygen release from HbO2. The resultant 15% additional oxygen partially compensates for low Hb blood levels in the anemic patients.