Phenotyping and long-term follow up of patients with hyper IgE syndrome

Introduction and objectives: Long-term follow up of patients with hyper IgE syndrome (HIES), as a primary immunodeficiency disorder, has been poorly investigated. This study describes common clinical and immunological features of patients with HIES in the last 10 years in Shiraz University of Medical Sciences, Shiraz, Iran. Methods and patients: In this cross-sectional study, the symptoms and medical records of 18 patients, who were diagnosed with HIES, were observed. Genetic and immunologic study was also performed. Results: Eighteen patients with the mean age of 13 years old were investigated. Ten patients were detected to have mutations in DOCK8 gene and autosomal recessive HIES (AR-HIES); and four patients were found with STAT3 mutation and autosomal dominant HIES (AD-HIES). So, 14 patients with known genetic results were considered for further data analysis. Food allergy, eczema, viral and skin infections were the major complications of AR-HIES patients. The major clinical complications of AD-HIES patients were pneumonia, skin infections and eczema. Food allergy and viral infection were significantly higher in DOCK8 deficient patients. The most common causes of hospitalization in both AR-HIES and AD-HIES patients were pneumonia, skin infections and sepsis. The most common cause of death was found to be sepsis. Conclusions; AD-HIES and AR-HIES cannot be differentiated only based on the clinical presentations. Genetic features are also necessary for better diagnosis. This study, summarizing the clinical, immunological and genetic information of the patients with AD-HIES and AR-HIES, may open a way for better diagnosis and management of HIES

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Methotrexate infusion followed by uterine artery embolisation for the management of placental adhesive disorders: a case series.

AIM: To evaluate the efficacy and safety of the uterine artery embolisation (UAE) in combination with methotrexate (MTX) for conservative management of placental adhesive disorders. MATERIAL AND METHODS: Patients with placental adhesive disorders (including accreta, increta, and percreta lesions) that were treated with UAE in combination with MTX were identified and were followed prospectively for outcomes including uterine preservation and complications. RESULTS: Twenty-six patients were identified who had the diagnosis of abnormal placenta implantation during this study. Fourteen patients were excluded because they were treated by a caesarean hysterectomy. Among remaining 12 patients, the successful uterine preservation observed in seven (58%) cases. Menstruation cycles returned in all successfully treated patients, although they did not have desire to get pregnant. Five (42%) patients underwent primary or delayed hysterectomy due to severe post-partum haemorrhage in three cases, and intestinal adhesion/peritonitis and secondary post-partum haemorrhage/sepsis in two patients, respectively. CONCLUSION: Although this interventional method is relatively successful in uterine preservation, the possibility of treatment failure cannot be ignored. Given that there are too few data regarding its efficacy and safety, this method should be reserved to patients who have a strong desire to maintain the uterus and their fertility, or if it is technically difficult to perform hysterectomy due to the extent of the invasion.

Phenotyping and long-term follow up of patients with hyper IgE syndrome.

INTRODUCTION AND OBJECTIVES: Long-term follow up of patients with hyper IgE syndrome (HIES), as a primary immunodeficiency disorder, has been poorly investigated. This study describes common clinical and immunological features of patients with HIES in the last 10 years in Shiraz University of Medical Sciences, Shiraz, Iran. METHODS AND PATIENTS: In this cross-sectional study, the symptoms and medical records of 18 patients, who were diagnosed with HIES, were observed. Genetic and immunologic study was also performed. RESULTS: Eighteen patients with the mean age of 13 years old were investigated. Ten patients were detected to have mutations in DOCK8 gene and autosomal recessive HIES (AR-HIES); and four patients were found with STAT3 mutation and autosomal dominant HIES (AD-HIES). So, 14 patients with known genetic results were considered for further data analysis. Food allergy, eczema, viral and skin infections were the major complications of AR-HIES patients. The major clinical complications of AD-HIES patients were pneumonia, skin infections and eczema. Food allergy and viral infection were significantly higher in DOCK8 deficient patients. The most common causes of hospitalization in both AR-HIES and AD-HIES patients were pneumonia, skin infections and sepsis. The most common cause of death was found to be sepsis. CONCLUSIONS: AD-HIES and AR-HIES cannot be differentiated only based on the clinical presentations. Genetic features are also necessary for better diagnosis. This study, summarizing the clinical, immunological and genetic information of the patients with AD-HIES and AR-HIES, may open a way for better diagnosis and management of HIES.

Cereals level and source effects on rumen fermentation, colostrum and milk properties, and blood metabolites in periparturient ewes.

Optimal type and dietary inclusion rates of cereal grains for periparturient sheep are unknown. The objective was to determine effects of feeding diets with high (H) v. low (L) levels of ground corn grain (CN) v. combined ground wheat and barley grains (WB) on intake, rumen fermentation, colostrum and milk properties, and blood metabolites of periparturient sheep. Twenty Afshari×Merino ewes were used in a completely randomized design study from 24 days prepartum through 21 days postpartum. Ewes were kept indoors in individual boxes and received once daily at 0900 h total mixed rations. Treatments were mixed rations containing either (1) H or (2) L concentrate based on either (1) 100% CN or (2) 50 : 50 ratio of ground wheat : ground barley grains in a 2×2 factorial arrangement. Each treatment group had five ewes including two twin-lamb ewes and three single-lamb ewes. Postpartal dry matter intake (DMI) increased by feeding high CN v. high and low WB, while high v. low CN improved postpartum DMI. The DMI during lambing tended to increase with the high v. low WB. Feeding CN v. WB, and feeding both CN and WB at L v. H level increased fecal pH. Postpartal rumen pH was lower with the high v. low WB (5.7 v. 6.2). Rumen concentrations of propionate were lower and of acetate were higher with L v. H grain levels. Increased dietary grain reduced urine pH for WB (7.24 v. 7.83) but not for CN (7.63 v. 7.52) prepartum. Colostrum properties, postpartal urine pH, lamb weight at birth and 21 days of age, and placental weight and expulsion time were unaffected. Milk yield increased and milk fat yield tended to increase by H v. L grain diets. Plasma glucose was increased by feeding high v. low WB, whereas CN v. WB tended to reduce peripartal plasma non-esterified fatty acids (NEFA) and increased insulin to NEFA ratio. In conclusion, more cereal grains can be included in periparturient sheep diets and CN instead of WB may be fed to periparturient sheep to improve energy status. Findings suggest opportunities to optimize periparturient ewe physiology and performance through feeding certain cereals and avoiding high levels of WB.

The morphological characters of the male external genitalia of the European hedgehog (Erinaceus Europaeus).

This study was conducted to depict anatomical characteristics of the penis of he-dgehog. Seven sexually mature male European hedgehogs were used. Following anaesthesia, the animals were scarified with chloroform inhalation. Gross penile characteristics such as length and diameter were thoroughly explored and measu-red using digital callipers. Tissue samples stained with haematoxylin and eosin and Masson's trichrome for microscopic analysis. The penis of the European hedgehog was composed of a pair of corpus cavernosum penis and the glans penis without corpus spongiosum penis. The urethra at the end of penis, protruded as urethral process, on both sides of which two black nail-like structures, could be observed. The lower part was rounded forming a blind sac (sacculus urethralis) with a me-dian split below the urethra. Microscopically, the penile bulb lacked the corpus spongiosum penis, but, corpus spongiosum glans was seen at the beginning of the free part. In the European hedgehog, entirely stratified squamous epithelium of penile urethra, absence of corpus spongiosum penis around the urethra and bilateral urethral glands are basically different compared with other mammals. This information is expected to contribute to comparative penile morphology as well as for testing phylogenic hypotheses and expanding knowledge about reproductive biology in this animal. (Folia Morphol 2018; 77, 2: 293-300).

The gross anatomy of the male reproductive system of the European hedgehog (Erinaceus Europaeus).

Hedgehogs are small spiny-coated insectivores. Due to their low body weight, calm character, and easy maintenance, they are kept as pets. It is therefore worthwhile to care about hedgehogs' health problems and to provide pet owners with information about their reproduction. Moreover, it is necessary to be familiar with their anatomy so as to satisfy the need to improve nutrition and medical care, even surgery. This study was carried out on five adult male European hedgehogs euthanased in a chloroform chamber. The European hedgehog's oval testes are invisible in inguinal region because they have no true scrotal sac. The testes are located in the craniocaudal direction with dorsolateral epididymal attachments. The vesicular glands, the European hedgehog's largest accessory sex glands, are lobulated structures containing dorsomedial and ventrolateral parts on each side. The prostate is an oval gland with right and left lobes. The paired bulbourethral glands are laid on the ischiocavernosus muscle. Histologically the vesicular, prostate gland ducts and ductus deferens as well as urethra separately were discharged in a diverticlum at the level of the pelvic urethra end. A sigmoid flexure exists in the proximal part of shaft body of the penis. There are two retractor penile muscles. In dorsal end of the penile glans, there is a small urethral process with two nail- -like, needle-shaped structures. They are on both sides of the urethral process. Furthermore, there are two intromittent sacs (Sacculus urethralis) in the ventral part of the end of the penis. (Folia Morphol 2018; 77, 1: 36-43).

Promising gene delivery system based on polyethylenimine-modified silica nanoparticles.

This article reports on the synthesis and full characterization of innovative silica-based nanoparticle composed of fumed silica as a core decorated with polyethylenimine (PEI) with different molecular weights (25, 10 and 1.8 kDa). Wide range of analytical, spectroscopic, and microscopic methods (TEM, DLS, ζ potential, elemental analysis (EA), TNBS and FTIR) were used to characterize the nanoparticles. Furthermore, transfection efficiency of these nanoparticles as non-viral vector was examined. The silica-PEI conjugates retained both the ability of PEI to fully condense plasmid DNA at low N/P ratios and suitable buffering capacity at the endosomal pH range. PEI-functionalized silica remarkably enhanced EGFP-N1 gene expression in murine neuroblastoma (Neuro-2A) cells up to 38 folds compared to PEI 25 kDa. Meanwhile the results of the cytotoxicity assays indicated that these silica-PEI conjugates have acceptable level of viability.

Histological concordance in familial central nervous system tumors: Evidence from nationwide Swedish Family-Cancer Database.

BACKGROUND: Published studies have shown that familial risks in the primary central nervous system (CNS) tumors are usually histology-specific. If genetic factors indeed determine tumor histology it would be expected that histological types would agree between affected first-degree relatives (FDRs). MATERIAL AND METHODS: This study was conducted using the nationwide Swedish Family-Cancer Database. FDR pairs were defined where both of them had the same histological subtype of CNS tumor. The histological concordance was determined using kappa agreement test. RESULTS: We identified 858 familial patients (333 parent-offspring pairs, 97 sibling pairs) with primary CNS tumors. Proportion of spinal hemangioblastoma out of all familial hemangioblastomas (21%) was significantly higher than that in sporadic patients (7%; P=0.001). The highest kappa value was found for hemangioblastoma among parent-child pairs (kappa=86%, 95% CI: 74-98%). There was a moderate agreement for concordant neurinoma among father-daughter pairs (kappa=48% 95% CI: 15-81%). Low grade glioma showed significant agreement among mother-daughter (kappa=33%, 95% CI: 9-57%) and father-daughter pairs (kappa=39%, 95% CI: 11-67%), but not in mother-son (kappa=10%, 95% CI: -13% to 32%) and father-son pairs (kappa=9%, 95% CI: -1% to 40%). There was histological agreements for meningioma in mother-offspring (kappa range=20-27%) but not in father-daughter (kappa=14%, 95% CI: -8% to 35%) and father-son pairs (kappa=9%, 95% CI: -12% to 30%). CONCLUSIONS: Our findings suggest that shared genetic risk factors between family members could lead to specific histological types in the familial CNS tumors, especially in hemangioblastoma and neurinoma. Our data may also suggest interactions between sex hormone and some genes contributing to familial meningioma and low grade glioma.

Reproductive biology of largescale tonguesole Cynoglossus arel in coastal waters of Bandar Abbas, Persian Gulf, Iran.

The objectives of this study were to determine the reproductive cycle of largescale tonguesole Cynoglossus arel, a commercially valuable flatfish species, in coastal waters of Bandar Abbas, along the south coast of Iran in the Persian Gulf. From October 2009 to September 2010, 905 fish were collected in monthly samples, and their length, weight, sex, gonad weight, and maturity status recorded. These data revealed that ovary weight in females is low from July to September, then increases to a peak in February followed by a decrease, indicating that the peak spawning season is from February to March with some spawning lasting until June. Males showed a corresponding seasonal pattern in testis weight, although with much less pronounced seasonal differences than gonad weight in females. Five maturity classes were described based on ovarian and testicular histology, corresponding with macroscopic analysis of gonads. The spawning season in C. arel is prolonged, similar to several other tropical flatfish species, and larger adult females tended to have an even more prolonged spawning period than smaller, presumably younger adult females. Combined, our results indicated that C. arel is a winter-to-spring batch spawner with an asynchronous type of ovarian development.

The effect of supplementation of calcium, vitamin D, boron, and increased fluoride intake on bone mechanical properties and metabolic hormones in rat.

Evidence indicates that optimal nutrition plays a role in bone formation and maintenance. Besides major components of mineralization such as calcium, phosphorus, and vitamin D, other nutrients like boron and fluoride have beneficial role, too. In this study, 34 male Wistar rats were divided into five groups: control diet, fluoride, fluoride + boron, fluoride + calcium + vitamin D, and fluoride + boron + calcium + vitamin D. Boron equal to 1.23 mg, calcium and vitamin D equal to 210 mg + 55 IU and fluoride equal to 0.7 mg/rat/day was added to their drinking water for 8 weeks. Plasma blood samples and bones were collected. Findings are evidence that fluoride + boron intake revealed significant positive effects on bone mechanical properties and bone metabolic hormones. These findings suggest that combined intake of these two elements has beneficial effects on bone stiffness and breaking strength comparing to even calcium + vitamin D supplementation. This evidence dealing with health problems related to bone and skeletal system in humans should justify further investigation of the role of boron and fluoride with other elements in relation to bone.

Risk factor investigation for cardiovascular health through WHO STEPS approach in Ardabil, Iran.

OBJECTIVES: Reliable evidence is the keystone for any noncommunicable disease (NCD) prevention plan to be initiated. In this study we carried out a risk factor investigation based on the WHO Stepwise approach to Surveillance (STEPS). METHODS: The study was conducted on 1000 adults between 15 and 64 years of age living in Ardabil province, north-west Iran during 2006, based on the WHO STEPS approach to surveillance of risk factors for NCD. At this stage only the first and second steps were carried out. Data were collected through standard questionnaires and methods analyzed using STATA version 8 statistical software package. RESULTS: 29.0% of men and 2.6% of women were current daily tobacco smokers. The mean number of manufactured cigarettes smoked per day was 18.9 among current daily smokers. Smoking was most prevalent among men of low-income families and those of lower education. The mean body mass index (BMI) was 26.6 kg/m(2), and was significantly correlated with systolic blood pressure. 58.9% were overweight or obese; 18.0% had raised blood pressure and 3.7% had isolated systolic hypertension. The mean number of servings of fruit consumed per day was 1.1; 33.1% had low levels of activity. Combined risk factor analysis showed that 4.1% of participants were in the low-risk group (up to 5.1% among men and 3.2% among women). Those in the high-risk group comprised 25.6% in the 25- to 44-year age group and 49.7% in the 45- to 64-year age group. Mean BMI increased by age in both sexes at least at the first three decades of adult life. CONCLUSION: Based on observed status of risk for cardiovascular health, burden of cardiovascular diseases is expected to increase if an effective prevention strategy is not undertaken.

A new radiopharmaceutical compound (131I-PR81) for radioimmunotherapy of breast cancer: labeling of antibody and its quality control.

PR81 is a monoclonal antibody that binds with high affinity to MUC1, which is over expressed on breast and other tumors. The objective of this study was to evaluate the application of this antibody against MUC1 as a radioimmunotherapeutical agent. Monoclonal antibody (PR81) against MUC1 was prepared, characterized, purified, and labeled with 131I. The immunoreactivity of radiolabeled mAb PR81with MUC1 (the native protein), BSA-P20 (a 20 amino acid corresponding the tandem repeat of MUC1) and MCF7 cell line were performed by RIA. In vitro stability of radiolabeled mAb in human serum was determined by thin layer chromatography (TLC). Cell toxicity and in vitro internalization studies were performed with the MCF7 cell line, and the tissue biodistribution of the radioiodinated PR81 was evaluated in normal BALB/c mice at 4, 24 and 48 hrs. The tumor imaging was performed in BALB/c mice with breast xenograft tumors at 24 and 72 hr after the complex injection. The labeling efficiency was found to be 59.9% ± 7.9%. MAb-131I conjugates showed high immunoreactivity towards MUC1 protein, BSA-P20 and MCF7 cell line. In vitro stability of the labeled product in human serum was found to be more than %50 over 24 hr. Cell toxicity and in vitro internalization studies showed that the mAb-131I conjugate inhibited 80% growth of the MCF7 cultured cell lines in vitro in a high concentration and up to %60 of the conjugate internalized after 24 h. Biodistribution studies were performed in normal BALB/c mice at 4, 24 and 48 hrs post-injection and no important accumulation was observed in vital organs. The tumors were visualized with high sensitivity after 24 and 72 hr in radioimmunoscintographical studies. These results show that the new radiopharmaceutical may be considered as a promising candidate for therapy of breast cancer.

Peritoneal retention of liposomes: Effects of lipid composition, PEG coating and liposome charge.

In the treatment of peritoneal carcinomatosis, systemic chemotherapy is not quite effective due to the poor penetration of cytotoxic agents into the peritoneal cavity, whereas intraperitoneal administration of chemotherapeutic agents is generally accompanied by quick absorption of the free drug from the peritoneum. Local delivery of drugs with controlled-release delivery systems like liposomes could provide sustained, elevated drug levels and reduce local and systemic toxicity. In order to achieve an ameliorated liposomal formulation that results in higher peritoneal levels of the drug and retention, vesicles composed of different phospholipid compositions (distearoyl [DSPC]; dipalmitoyl [DPPC]; or dimiristoylphosphatidylcholine [DMPC]) and various charges (neutral; negative, containing distearoylphosphatidylglycerol [DSPG]; or positive, containing dioleyloxy trimethylammonium propane [DOTAP]) were prepared at two sizes of 100 and 1000nm. The effect of surface hydrophilicity was also investigated by incorporating PEG into the DSPC-containing neutral and charged liposomes. Liposomes were labeled with (99m)Tc and injected into mouse peritoneum. Mice were then sacrificed at eight different time points, and the percentage of injected radiolabel in the peritoneal cavity and the tissue distribution in terms of the percent of the injected dose/gram of tissue (%ID/g) were obtained. The ratio of the peritoneal AUC to the free label ranged from a minimum of 4.95 for DMPC/CHOL (cholesterol) 100nm vesicles to a maximum of 24.99 for DSPC/CHOL/DOTAP 1000nm (DOTAP 1000) vesicles. These last positively charged vesicles had the greatest peritoneal level; moreover, their level remained constant at approximately 25% of the injected dose from 2 to 48h. Among the conventional (i.e., without PEG) 100nm liposomes, the positively charged vesicles again showed the greatest retention. Incorporation of PEG at this size into the lipid structures augmented the peritoneal level, particularly for negatively charged liposomes. The positively charged PEGylated vesicles (DOTAP/PEG 100) had the second-greatest peritoneal level after DOTAP 1000; however, their peritoneal-to-blood AUC ratio was low (3.05). Overall, among the different liposomal formulations, the positively charged conventional liposomes (100 and 1000nm) provided greater peritoneal levels and retention. DOTAP/PEG100 may also be a more efficient formulation because this formulation can provide a high level of anticancer drug into the peritoneal cavity and also can passively target the primary tumor.

Effect of liposome size on peritoneal retention and organ distribution after intraperitoneal injection in mice.

Peritoneal carcinomatosis is a serious concern when treating digestive or ovarian tumors. Treatment with systemic chemotherapy suffers from poor penetration of cytotoxic agents into the peritoneal cavity and is not quite effective. Local delivery of drugs, especially as controlled-release delivery systems like liposomes, could provide sustained and higher drug levels and reduce systemic toxicity. In order to investigate the effect of liposome size on peritoneal retention, liposomes composed of distearoylphosphatidylcholine and cholesterol (DSPC/CHOL, molar ratio 2:1) were prepared at four sizes of 100, 400, 1000 and 3000 nm. Subsequently, these liposomes were labeled with (99m)Tc complex of hexamethylpropyleneamineoxime ((99m)Tc-HMPAO) and injected into mouse peritoneum. Then, mice were sacrificed at eight different time points and the percentage of injected radiolabel in the peritoneal cavity and the organ distribution in terms of percentage injected dose/gram tissue (%ID/g) were obtained. Results showed that the free label ((99m)Tc-HMPAO) was cleared very rapidly from the cavity so that after 5 min and 7h only 6.89+/-2.51% and 0.91+/-0.51% of the injected dose was recovered, respectively. However, for the liposomal formulations, this recovery value ranged from 8.47+/-1.62% to 29.99+/-12.06% at 7h. Peritoneal retention of the vesicles was increased with their size, and the highest retention rate was obtained with 1000 nm liposomes with an AUC value 15.51 times that of (99m)Tc-HMPAO. In blood, as expected, 100 nm liposomes showed much higher levels because of their greater stability. Their greater blood concentration also caused increased levels in the heart and kidneys, although their organ to blood AUC ratio was the lowest. Overall, among the different sized neutral liposomes investigated, the 1000 nm vesicles seemed to be the most optimal, achieving a greater peritoneal level and retention.

The radiotracer (99m)Tc-MIBI is not genotoxic for human peripheral blood lymphocytes at diagnostic radioactive dose.

The radiotracer technetium-99m methoxyisobutyl isonitrile ((99m)Tc-MIBI) has been widely used for myocardial blood flow imaging. We investigated the genotoxicity of (99m)Tc-MIBI in cultured human lymphocytes at the same concentration used in patients. Radioactivity doses were determined in whole blood at 5 min post-injection of 20 mCi (99m)Tc-MIBI in patients. Subsequently, whole blood of human volunteers was incubated with 1, 2.3, 4 or 8 microCi (99m)Tc-MIBI. After a 30-min incubation, the lymphocytes were stimulated with a mitogen to assay for micronuclei in cytokinesis-blocked binucleated cells. The frequency of micronuclei in samples treated with this radiopharmaceutical up to 2-fold (8 microCi) the concentration of (99m)Tc-MIBI normally found in the blood of patients was not more than in control lymphocyte cultures. We concluded that there is no increased induction of micronuclei in lymphocytes incubated with (99m)Tc-MIBI at the radioactivity doses used for diagnostic purposes.

Association of Helicobacter pylori infection with cardiac syndrome X.

INTRODUCTION: Cardiac syndrome X (CSX) includes chest pain, positive exercise stress test and/or radionuclide test for ischaemia and normal coronary angiography. There is no obvious aetiology for this syndrome. Some mechanisms such as endothelial dysfunction and oestrogen deficiency have been invoked. In this study, we surveyed the association of Helicobacter pylori (HP) infection with cardiac syndrome X. METHODS: HP infection was detected by urea breath test (UBT) in patients with cardiac syndrome X, and compared with a sex- and age-matched control group. Patients with dyspepsia and coronary spasm were excluded. Statistical analysis was carried out using chi-square test. RESULTS: 40 patients (29 females and 11 males) with cardiac syndrome X aged between 30 and 65 years (mean 45.51 +/- 5.03 years) were compared with a control group (28 females and 12 males) aged between 31 and 64 years old (mean 44.93 +/- 5.16 years). 95 percent of patients were HP infected, while only 47.5 percent of members of the control group were infected (p-value is less than 0.001). CONCLUSION: Considering the high prevalence of HP infection in patients with CSX in our sample and probable causative effect of chronic infection in vascular diseases, we believe that there is a probable role for HP infection in the pathogenesis of CSX.

Direct labelling of octreotide with 99mTc: effect of different concentration of reducing agents and amount of sodium pertechnetate on radiolabelling efficiency.

Octreotide, a synthetic analog of natural hormone somatostatin, was labelled with 99mTc. Labelling was accomplished by reduction of the cysteine bridge, which provided sulfhydryl groups for chelating with 99mTc. Sodium ascorbate and sodium dithionite in different concentrations were used as reducing agents. Different amounts of sodium pertechnetate were used for labelling of peptide. When the mass ratio of peptide and sodium ascorbate was 1:100 and the final concentration of dithionite in the labelling vial was 0.2-0.4 microg/microl with 0.18-1.48 GBq sodium pertechnetate more than 80% radiolabelling efficiency was confirmed by RP-HPLC, ITLC-SG and C18 Cartridge analysis. The stability of the 99mTc-peptide bond was evaluated by human serum challenge and that showed the stability was 90% after 4h.

Polymerase chain reaction of bacterial genomes with single universal primer: application to distinguishing mycobacteria species.

The polymerase chain reaction with a single universal primer (UP-PCR) was applied to bacterial strains and mycobacteria isolates alongside conventional methods. A universal protocol of preparation of PCR samples from cultures representing Escherichia coli, Enterobacter aerogenes, Serratia marcescens, Staphylococcus aureus, Streptococcus pyogenes, Klebsiella pneumoniae, Mycobacterium tuberculosis, Mycobacterium bovis, and several non-tuberculous mycobacteria was found to be reproducible and efficient with these organisms. The bands of UP-PCR products observed in an agarose gel after electrophoresis were species-specific and provided an efficient means of distinguishing bacterial species. The applicability of this approach to mycobacteria identification was assessed by comparing the DNA bands obtained for different strains. Three reference strains and 22 clinical isolates of M. tuberculosis and M. bovis produced very similar DNA banding patterns. They comprised a triplet of prominent and several minor fragments within the 200-500 base pair (bp) size range and were the easiest to interpret. The DNA profiles of unrelated mycobacteria clearly differed from each other when subjected to electrophoretic analysis and correlated well with results of culture method. The method provides a real promise of its application in clinical studies as a simple assay for distinguishing between tubercle bacilli.