Bone marrow micro-metastases in clinically stage I-III HER2/Neu positive breast cancer

Patients with breast cancer should have a favorable outcome when diagnosed early. Occult distant metastases, especially in the Bone Marrow [BM] was claimed to be responsible for tumor relapse in certain cases, especially in HER-2/neu expressing tumors. Such an occult metastases may be detected by PCR techniques for a BM aspirate, which is the aim of this study. Between May 2004 and May 2005, PCR technique was used to determine the tumor HER-2/neu status of 37 patients, attending Suez Canal University Hospital for the treatment of stage I-III breast cancer. BM and peripheral blood of those positive patients were examined by the same technique for the presence of micro-metastases. Correlation between tumor characteristics and the HER-2/neu status was done. Fifteen patients were tumor HER-2/neu positive, which correlated with stage and number of positive lymph nodes. BM micro-metastases were detected in 7 patients, with no correlation to the tumor characteristics. Additionally, HER-2/neu positive tumor cells were detected in the peripheral blood of 6 patients. BM micro-metastases can be detected in HER-2/neu positive non-metastatic breast cancer that may be predicted by the peripheral blood not the primary tumor characteristics

Real-time PCR and flow cytometry in detection of cyclospora oocysts in fecal samples of symptomatic and asymptomatic pediatrics patients

The magnitude of Cyclospora oocysts excretion in relation to infection intensity among cyclosporiasis patients was assessed using flow cytometry and quantitative real-time PCR [RT-PCR]. Oocysts from stool samples of 25 [14.8%] gastro-intestinal symptomatic pediatrics patients [169] and of 10 [2.8%] asymptomatic gastrointestinal ones [350] were identified by modified Ziehl-Neelsen [MZN] and modified Acid Fast Trichrome [MAFT] staining methods and confirmed by its auto-fluorescent characterizations. Also, 10 infants with negative stool samples were selected as controls. The intensity of infection was calculated as number of oocysts/200 microscopic filed with immersion 400. Flow cytometry and RT-PCR assessed relation between symptoms and oocysts excretions compared to MZN and MAFT. The infection severity in symptomatic patients were identified by MZN and MAFT as mild [16%], moderate [24%] and severe [60%]. All asymptomatic patients had mild infection. Flow cytometry was done for stool samples and 100% Cyclospora oocysts were in symptomatic and asymptomatic patients. None was detected in controls, RT-PCR was done for stools with both a species-specific primer set and dual fluorescent labeled Cyclospora cayetanensis hybridization probe by unique regions of 18S rRNA gene sequence. DNA of C. cayetanensis was in 100% of symptomatic and asymptomatic patients and in 20% of controls. In repetitive examination of stools Cyclospora oocysts were neither detected by staining nor flow cytometry. Based on oocysts counts, no differences were found between flow cytometry and RT-PCR in compared to staining methods

Evaluation of mycobacterium growth indicator tube and gen-probe amplified mycobacterium tuberculosis direct test for rapid diagnosis of mycobacterium tuberculosis in sputum specimens

The Mycobacterium Growth Indicator Tube [MGIT] and Gen-Probe Amplified Mycobacterium tuberculosis Direct [AMTD] test were evaluated using 52 respiratory clinical specimens collected from suspected pulmonary tuberculosis patients. Microbiological culture on Lowenstein-Jensen medium was used as the reference method. The 52 sputum specimens, 29 [55.8%] acid fast smear positive and 23 [44.2%] acid fast smear negative, were cultivated in liquid MGIT and on solid U media. The recovary rate of MTB by MGlT was [80.8%]. The Kappa coefficient calculation showed a very good agreement [Kappa=0.83] between the 2 tests but there was no statistical difference in the sensitivity of detection of MTB between MGIT and Li methods [P>0.05]. There was a statistically significant difference [P<0.05] between the 2 methods as regards the mean time of growth detection. The mean [range] time of detection of MTB was 12.5 [5 to 30] and 19.5 [14-30] days with MGIT and Li, respectively. The recovery rate of MTB 1mm both media in combination [MGIT plus LJ] was higher [84.6%] than the recovery rate on each media separately and the difference between the 2 methods was not statistically significant [P>0.05]. Regarding the detection of MTB from smear positive and smear negative sputum, it was found that MOLT and U detected 93% and 86.2% respectively of the 29 smear positive specimens whereas they detected 65.2% and 60.8% respectively of the 23 smear negative specimens and there was no statistical difference between the 2 methods in relation to the smear type. The recovery rate of MTB by AMTD was 71%. The assay had sensitivities of 100 and 85.7% for acid fast smear positive and negative specimens, respectively. The specificity of the assay was 100% for both types of smears. Our overall results for all specimens, regardless of smear status, showed a sensitivity of 94.8%, a specificity of 100%, a positive predictive value of 100% and a negative predictive value of 86.6%. In conclusion, Both MGIT and AMTD are reliable methods for rapid diagnosis of pulmonary tuberculosis but they should not be used instead of a solid medium; rather, they should be used in addition to it

Incidence of cow milk allergy in colicky baby

There is a strong relation between infantile colic and the early introduction of cow milk [raw or artificial] to the infants in the first few months of life. Cow milk proteins are responsible for the pathological changes which precipitate the different symptoms of allergy including colic. This study aimed to estimate the prevalence of cow milk allergy [CMA] in the infants suffering from repeated attacks of colic and to determine the adverse effects of CMA. 30 infants suffering from repeated attacks of colic were included in this study. They subjected to full history taking especially the dietary history, also were subjected to clinical examination with special attention to growth development and other signs of allergy. Blood sample was taken to test for CMA including, blood eosinophilic count, total IgE and S-IgE levels to cow milk proteins in the serum. In addition, patients were subjected to an open challenge test to cow milk, which is the gold standard for diagnosis of CMA. 13 children out of the 30 were CMA +ve i-e 43.3% of the studied patients had CMA diagnosed by challenge test. Study of the dietary history revealed that the total and mean duration of exclusive breastfeeding were significantly shorter in CMA+ve than in CMA-ve patients[p < 0.05 for both].in addition the age of first introduction of CM was significantly earlier in CMA+ve compared to CMA-ve patients[p < 0.05]. However, type of milk [raw or artificial or diary product as yogurt], showed no significant difference to the diagnosis of CMA in our study group. Other allergic manifestations were recorded in CMA+ve patients especially infantile eczyma. None of our patients had peripheral eosinophilia. High levels of total IgE and specific IgE were detected in 46.2% and 36.6% respectively. Specific IgE showed a higher specificity than total IgE [100% vs 52.9%].High levels of both total and Specific IgE were significantly more common in CMA+ve patients compared to CMA-ve patients. In addition, IgE - mediated reactions accounted for 46.2% of CMA+ve patients. CMA is a common finding in infants suffering from colic in the first months of life. Early introduction of cow's milk in the first year of life is frequently associated with increase risk of CMA. Testing total and specific IgE for cow milk is a useful tool for the diagnosis of suspicious cases. Challenge test is still the gold standard for the diagnosis of CMA. Although Exclusive breastfeeding in the first 6 months of age is not absolutely protective against all causes of colic, it is still obligatory for the prevention of the development of food allergy