13910C>T and 22018G>A LCT gene polymorphisms in diagnosing hypolactasia in children.

Background: Primary lactose intolerance (PLI) is a gradual decrease of lactase activity that usually manifests at the age of 1-5 years. It has been proved that PLI is related to a single-nucleotide polymorphism of the lactase (LCT) gene. Objective: An evaluation was performed on the usefulness of genetic tests in detecting LCT 13910C>T and 22018G>A polymorphisms in diagnosing lactose intolerance in children. Methods: The study group included 99 children aged from 2 months to 16.5 years with different digestive tract symptoms. In all patients a hydrogen breath test (HBT) was conducted and blood samples were collected to determine LCT polymorphisms. PLI was defined as the presence of the 13910CC and/or 22018GG polymorphism in patients with a positive HBT result. Results: In the group younger than 6 years, no statistically significant correlation was observed between the 13910CC and/or 22018GG LCT polymorphisms and HBT result. In the group of children older than 6, a statistically significant correlation between the 13910CC (p = 0.0011) and 22018GG (p = 0.003) LCT polymorphisms and HBT result was detected. Conclusions: In children older than 6, the result of genetic testing based on LCT 13910C>T and 22018G>A polymorphisms may diagnose lactose intolerance.

Differentiation of Clavibacter michiganensis subsp. sepedonicus using PCR melting profile and variable number of tandem repeat methods.

The potato phytopathogen Clavibacter michiganensis subsp. sepedonicus (Cms) is a causative agent of bacterial ring rot, which is a serious threat to crops. In EU member countries, Cms is subject to quarantine and has to be combated. The knowledge about the transmission of C. michiganensis strains is limited due to a lack of methods which could be used for epidemiological analysis. In this study, PCR melting profile (PCR MP) and variable number tandem repeat methods were used in Cms epidemiological analysis for the first time. PCR MP was based on the melting temperature analysis of BamHI restriction fragments of chromosomal DNA. Respectively, for the variable number tandem repeat (VNTR) method, six loci were identified and used in the differentiation of Cms isolates. PCR MP was used for 93 Cms isolated in Poland. Both PCR MP and VNTR methods were used for the differentiation of 47 Cms strains in this collection. Both these methods were found to be useful for the epidemiological analysis of Cms. SIGNIFICANCE AND IMPACT OF THE STUDY: The potato phytopathogen, Clavibacter michiganensis subsp. sepedonicus (Cms), is a serious threat to crops and lead to significant economic losses. The only way to control and eliminate the disease caused by this pathogen is the use of certified seed potato and strict quarantine of infected fields. Here, for the first time, two molecular typing methods (PCR melting profile (PCR MP) and variable number tandem repeat (VNTR)) were evaluated in respect of their potential in differentiation of Cms isolates. As a result, we obtained characteristic profiles of DNA fragments (PCR MP) and numeric patterns (VNTR), which enable the intraspecies genotyping of Cms strains confirming the effectiveness of PCR MP and VNTR methods in differentiation of Cms strains.

Development of a new ligation-mediated PCR method for the differentiation of Mycobacterium tuberculosis strains.

BACKGROUND: There is a need for rapid, inexpensive methods for analysing a limited number of Mycobacterium tuberculosis strains. The ligation-mediated polymerase chain reaction (LM-PCR) method appears to be sufficiently discriminative and reproducible to be considered as a molecular tool for the initial evaluation of hospital outbreaks, laboratory cross-contamination, and family or small community transmission. OBJECTIVE: To develop a new LM-PCR method based on PCR amplification of the 5'-flanking region of insertion sequence (IS) 6110 consisting of SalI/PvuII digestion of chromosomal DNA, ligation of a SalI linker and differentiation of IS6110-carrying restriction fragments by suppression subtractive hybridisation. DESIGN: The fast ligation amplification polymorphism (FLAP) method was applied in the analysis of 62 M. tuberculosis clinical isolates and compared with IS6110-restriction fragment length polymorphism (RFLP) and mycobacterial interspersed repetitive units-variable number of tandem repeat (MIRU-VNTR) analyses of the same strains. RESULTS: The sensitivity of FLAP was estimated at 0.25 ng/l. FLAP yielded 32 patterns among the 62 M. tuberculosis strains compared to respectively 28 and 36 patterns obtained using MIRU-VNTR and IS6110-RFLP. Its Hunter-Gaston discriminatory index value (0.973) is similar to that of MIRU-VNTR (0.966) and IS6110-RFLP (0.971). The specificity of the FLAP patterns was also confirmed. CONCLUSION: FLAP proved highly discriminating, sensitive and specific and could be a valuable molecular tool, especially for analysing a limited number of M. tuberculosis strains.

A phase II study of the combination of endocrine treatment and bortezomib in patients with endocrine-resistant metastatic breast cancer.

The majority of patients with hormone receptor-positive metastatic breast cancer die from disease progression despite different types of anti-hormonal treatments. Preclinical studies have indicated that resistance to anti-hormonal therapies may be the result of an activated NF-κB signalling pathway in breast cancer. Bortezomib is a proteasome inhibitor that blocks the NF-κB pathway. Recent pharmacodynamic and pharmaco-kinetic xenograft studies have shown that drug exposure may be a crucial factor for the efficacy of bortezomib in solid tumours. The aim was to investigate whether the addition of bortezomib to anti-hormonal therapy would result in regained antitumour activity in patients with progressive and measurable disease being treated with an endocrine agent. Clinical benefit was defined as patients obtaining stable disease, partial response or complete response after 2 cycles, lasting for at least another five weeks. Bortezomib was administered on days 1, 8, 15 and 22 of a 5-week regimen (1.6 mg/m2). Eight patients received an aromatase inhibitor and bortezomib, while one received tamoxifen and bortezomib. There were 3 grade 3 gastrointestinal toxicities. Median time to treatment failure was 69 days (range, 35-140). Two out of the 9 patients had stable disease for more than 10 weeks. Despite an effective target inhibition, suggested in peripheral blood mononuclear cells and available tumour samples, no objective antitumour responses were observed. Addition of a proteasome inhibitor to anti-hormonal therapy resulted in a clinical benefit rate of 22% in a limited number of patients with endocrine resistant and progressive metastatic breast cancer. The demonstrated proteasome inhibition in tumour tissue provides evidence that the lack of clinical responses is not attributed to deficient drug exposure.

Epidemiological analysis of Mycobacterium tuberculosis strains isolated in Lodz, Poland.

BACKGROUND: Mycobacterium tuberculosis is one of the most dangerous human pathogens. Molecular typing of M. tuberculosis has allowed better control of tuberculosis and, among other benefits, identification of genetic lineages among strains. OBJECTIVE: To test the potential of polymerase chain reaction (PCR) based methods for the epidemiological study of M. tuberculosis strains isolated from patients residing in a single city. DESIGN: We performed spoligotyping, mycobacterial interspersed repetitive units-variable number of tandem repeats (MIRU-VNTR) typing and insertion sequence (IS) 6110 restriction fragment length polymorphism (RFLP) analyses of 234 clinical strains of M. tuberculosis collected over 2 years from the Polish city of Lodz. RESULTS: Spoligotyping analysis revealed 84 spoligotypes with a shared international type and 50 unique spoligotypes. Subtyping via 15- and 19-loci MIRU-VNTR analyses revealed 154 patterns with 117 unique profiles, and 159 patterns with 126 unique profiles, respectively. Spoligotyping combined with MIRU-VNTR 15- and 19 loci analyses revealed 132 and 146 unique profiles, respectively. Overall, 96 strains clustered via MIRU-VNTR typing were used in IS6110-RFLP analysis. Complete congruence of patterns revealed by PCR-based methods was noted for 40 strains, of which 36 were isolated from epidemiologically linked patients. CONCLUSION: The combination of 15-loci MIRU-VNTR typing with spoligotyping is useful for primary analysis of M. tuberculosis strains; however, additional use of MIRU 23 should be considered. Strains clustered by PCR-based methods should be further analysed by IS6110-RFLP typing.

Studies on iodine content in daily diets, particularly elderly people's diets.

Poland represents moderate degree of severity of iodine deficiency. In 1997 a national program has been introduced of obligatory iodine prophylaxis including the iodination of household salt. In order to assess the extent of iodine consumption, the assessment were carried out on iodine content in average Polish diets with regard elderly people's diets. The aim of the study was also to analyse the composition of supplements allowed for trade in Poland, with regard to iodine content in these products. The assessment of iodine content in Polish diets was performed on the basis of the data of the household budgets and the data of iodine content in food products (without kitchen salt). The calculated iodine content in average Polish daily diets (adults and children) ranged from 40.4 microg to 50.7 microg. The iodine content in elderly people's diets was 75.5 microg/daily diets. In all studied diets the amount of iodine coming from food products, not taking into account iodinated salt, was insufficient for the realization of Polish RDA for this element. It shows that iodination of kitchen salt in Poland is necessary. Moreover on the Polish market exist supplements including iodine (20-200 microg iodine in tablet).

[Magnesium status in children and adolescents with celiac disease]. / Stan odzywienia magnezem u dzieci i mlodziezy z celiakia.

UNLABELLED: Magnesium deficiency has been reported in patients with classical coeliac disease. Classical coeliac disease has been recently very rare, but the frequency of the silent or latent form has increased. The aim of the study was to evaluate the magnesium status in patients with coeliac disease diagnosed according to ESPGAN criteria. 41 GFD(+) patients aged 6-18 years, who were on a gluten-free diet (GFD) for 2.8 to 17.3 years (mean 11 years); with normal villous structure and IgAEmA(-), and 32 GFD(-) patients aged 5-17 years, with villous atrophy and IgAEmA(+): 8--after 7/12-13/12 of gluten challenge, 4--with late onset of coeliac disease, 20--with silent coeliac disease. All of the children did not have any other disorders. Magnesium status was examined by using: an i.v. Mg-loading test (30 mmol/1.73 m2); Mg urinary excretion and Mg concentration in serum, erythrocytes, and in hair. Abnormal values in GFD(+) and GFD(-) patients were found in: Mg i.v. loading test (retention > 40%) in 20 vs 34%, serum Mg (< 0.7 mmol/l) in 7 vs 3%, erythrocytes Mg (< 1.8 mmol/l) in 20 vs 25%. The reversed statistically significant correlation was found between Mg retention and Mg urinary excretion (R = -0.293, p = 0.009). No other statistically significant correlations were found. CONCLUSION: The magnesium deficiency was present in all patients with classical coeliac disease, but only in 1/5 of patients with coeliac disease on a gluten-free diet and in 1/5 of patients with silent coeliac disease.

[Reasons for magnesium deficiency in children with coeliac disease]. / Przyczyny niedoboru magnezu u dzieci i mlodziezy z celiakia.

UNLABELLED: Magnesium (Mg) deficiency is often noted in patients with coeliac disease (CD). The aim of the study was the analysis of the reasons of this deficiency in children with CD, diagnosed according to ESPGAN criteria. MATERIAL: The study was performed on 41 patients aged 6-18 years adhering to strict gluten-free diet GFD(+) for mean 11 years, with normal small intestine mucosa, and IgAEmA(-), and on 32 patients aged 5-17 years on gluten containing diet, with classical CD, silent CD or after gluten challenge--GFD(-). In this group the villous atrophy of the small intestine and IgAEmA(+) were observed. In 18 of these patients Mg deficiency was found using Mg-loading test (30 mmol/1.73 m2). METHODS: The following parameters were analysed: type of the disease, observance of gluten-free diet, sex, and living place. Mg, Ca, Na, protein, fat, and dietary fiber intake was assessed using food frequency questionnaire method, and steatorrhea using faecal fat excretion (g/24 h). RESULTS: The frequency of Mg deficiency was similar in both sexes, occasionally in children from small towns (4.5%), and more often in children from big cities (31.5%), and village (34.4%). Dietary Mg intake below RDA was observed in 23% of children from GFD(+) group, in 19% from GFD(-) one, and in 17.6% in children with Mg deficiency. Insufficient Mg intake was found in 18.2% of children from small towns, in 17.6% from big cities, and in 12.5% from villages; Ca in 36.6%, 58.8%, and 59.3%, and protein in 18.2%, 35.3%, and in 34.4% respectively. In all groups of children high intake of fat and Na was observed. Dietary fiber intake was within the recommended values. All children with classical CD had increased fat excretion (mean 25.9 g/24 h), in other patients it was within normal values [GFD(+) mean 1.95 g/24 h, in GFD(-) without diarrhoea 1.7 g/24 h. CONCLUSIONS: Magnesium deficiency in children with CD depends on the form of the disease, adhering to GFD, diarrhoea with steatorrhea, and/or low Mg intake with the diet.

Polyamines in cow's and sow's milk.

There were considerably interspecies, individual, lactation and age related variabilities in the concentration of spermidine and spermine in cow's and sow's milk. Concentration of spermidine was similar in cow's and sow's milk, whereas spermine level was higher in sow's milk. The level of spermine was higher than spermidine throughout the lactation in sow's milk, whereas spermine in cow's milk was secreted only at the beginning of lactation (collostrum and milk of the first month of lactation). The peak of spermidine and spermine concentration occurred in the collostrum and milk between the 1st and 3rd week of lactation in cow and sow, respectively. There was a significant positive relationship between milk yield and spermidine secretion in cow as well as between the number of piglets and milk spermidine concentration in sow.

Diagnostica--a Bayesian decision-aid system--applied to hypertension diagnosis.

"Diagnostica" is a Bayesian statistical tool designed to collect and store the patient's data, suggest a diagnosis, and explain the decision in terms of density distributions. The program is written in C language on MacIntosh support. It is described using the case study of differential diagnosis between essential and secondary hypertensions (i.e., fibrodysplastic renal artery stenosis, atheromatous renal artery stenosis, Conn's syndrome, nephropathy and pheochromocytoma). Seventeen experimental parameters were taken into consideration, all of them available during the first medical examination. The density distributions of all items were established from the ARTEMIS experimental database. Both a priori probabilities of different types of hypertension and loss coefficients are taken into account in the calculations. Diagnostica can be used in a "make diagnosis" mode or in an "edition" mode. In the first case it can serve a physician in everyday practice; in the second it becomes a tool for medical research.

[Nutritive value of daily food rations reproduced in different regions of the country. IV. Copper, zinc and manganese levels]. / Wartosc odzywcza calodziennych racji pokarmowych odtwarzanych w kilku regionach kraju. CZ. IV. Zawatosc miedzi, cynku, manganu.

The content of copper, zinc and manganese in daily diets reconstructed in the laboratory in 1988 on the basis of data on the yearly food consumption in 1986 of two social groups (manual and mental workers) with medium income was determined. The diets were prepared for four regions of the country (Warszawa, Lublin, Poznan, Wroclaw). It was found out that the content of copper and zinc in the diets in 1986 and the realisation of recommended dietary allowances for these minerals were lower than in 1973, 1980 and 1981. The degree of realization of the recommended intake of zinc and copper in diets from 1986 was on average 67% and 45% respectively. The content of manganese in the diets was in the range of recommendation. The content of microminerals in the diets was usually similar in the all studied regions of the country.

[Nutritive value of daily food rations prepared in different regions of the country. III. The levels of calcium, phosphorus, magnesium, iron and potassium]. / Wartosc odzywcza calodziennych racji pokarmowych odtwarzanych w kilku regionach kraju. Cz. III. Zawartosc wapnia, fosforu, magnezu, zelaza i potasu.

Studies on the content of macrominerals in daily diets reconstructed in 1988 on the basis of the analysis of family budgets in 1986 carried out by the Central Statistical Bureau were carried on. Two social groups i.e. manual and mental workers with medium income were considered. The diets were prepared for 5 regions (Warszawa, Lublin, Olsztyn, Poznan, Wroclaw). According to the studies the requirements for calcium and magnesium were met in about 70% and those for iron in about 84%. The content of potassium in the diets was in the range of recommended allowances while the phosphorus exceeded the allowances by about 20-30%. The comparison of the presently studied diets with the ones from 1973, 1980, 1981 showed a lower degree of realization of the recommended intake of calcium and magnesium in 1986. The content of studied minerals in the diets was usually similar in all five regions.