The downstream drifting of larvae of Dixa (Diptera: Dixidae) in two stony streams

SUMMARY. Larvae of Dixa , especially D. puberula Loew., were a significant component of the invertebrate drift in the Walla Brook in southwest England, the Wilfin Beck and the River Duddon in the English Lake District, and the River Estibère in the French Pyrenees. The drifting of the larvae increased markedly at night and showed a definite diel periodicity with maximum numbers usually in the early hours of the night. Seasonal peaks in the density of larvae in the drift frequently occurred in months when adults and especially pupae were present. Most drifting larvae were in the fourth and final instar, and their drift rate usually increased when they were searching for suitable pupation sites.     

A comparative study of four dredges used for sampling benthic macroinvertebrates in rivers

SUMMARY. After considering the large number of dredges described in the literature, four light-weight dredges were chosen for manual operation from a small boat or the bank: Irish triangular dredge, small Fast dredge, medium-sized and large Naturalist's dredges. The dredges were tested in a series of trials at three sites in two rivers. A stratified random sample (number of sampling units, n = 5) was taken at each site and the modal particle sizes at sites 1–3 were 1–2 mm (fine gravel), 64–128 mm (larger stones) and 128–256 mm, respectively. The dredges usually took a similar range of stone sizes at each site but the design of the Fast dredge excluded larger stones (>16 mm). The Irish dredge sometimes failed to operate correctly. Variations in the volume of substrata taken with each dredge were large, both between sampling units in the same sample and between samples. The latter differences were partially due to the increase in the modal size of the stones, especially between sites 1 and 2, the different sampling areas of the dredges and the depth of penetration into the substratum. Penetration depth was probably greatest for the two Naturalist's dredges, smaller for the Fast dredge and smallest for the Irish dredge. In field trials, the relative abundances of major taxa were similar for most dredges at each site; major exceptions were the Fast dredge at site 2 and the Irish dredge at site 3. There was a high variability between sampling units in the same sample and therefore a lack of precision in the estimates of the mean number of invertebrates per sample. Therefore, the dredges cannot be used as quantitative samplers for the estimation of population density. Their adequacy as qualitative samplers for the estimation of total number of taxa per sample varied considerably and maximum estimates of their efficiencies for a small sample (n= 5) were <40% for the Irish and Fast dredges, >57% for the medium-sized Naturalist's dredge and >76% for the large Naturalist's dredge. There was a clear relationship between the number of taxa and the number of invertebrates taken at each site and this relationship was well described by a power law with an exponent within the range 0.18–0.53. The number of sampling units in the sample had no significant effect on the power-law equations for each site. The power-law equation was very similar for most of the dredges at each site, the only major exception being the Fast dredge at site 1. The implications of this relationship are discussed.     

Intratumor heterogeneity of biomarker expression in breast carcinomas

Small biopsy samples are used increasingly to assess the biomarker expression for prognostic information and for monitoring therapeutic responses prior to and during neoadjuvant therapy. The issue of intratumor heterogeneity of expression of biomarkers, however, has raised questions about the validity of the assessment of biomarker expression based on limited tissue samples. We examined immunohistochemically the expression of HER-2neu (p185^erbB-2), epidermal growth factor receptor (EGFR), Bcl-2, p53, and proliferating cell nuclear antigen (PCNA) in 30 breast carcinomas using archived, paraffin embedded tissue and determined the extent of intratumor heterogeneity. Each section was divided into four randomly oriented discrete regions, each containing a portion of the infiltrating carcinoma. For each tumor, the entire lesion and four regions were analyzed for the expression of these markers. Scores of both membrane and cytoplasmic staining of HER-2neu and EGFR, scores of cytoplasmic staining of Bcl-2, and scores of nuclear staining of both p53 and PCNA were recorded. The intensity of staining and the proportion of immunostained cells were determined. A semiquantitative immunoscore was calculated by determining the sum of the products of the intensity and corresponding proportion of stained tumor cells. We analyzed both invasive (IDC) and in situ (DCIS) carcinomas. The Wilcoxon signed-rank test was used for paired comparisons between overall and regional immunoscores and between overall and regional percentages of stained cells. Spearman's correlation coefficients were used to assess the level of agreement of overall biomarker expression with each of the regions. Generalized linear models were used to assess overall and pair-wise differences in the absolute values of percent changes between overall and regional expression of biomarkers. For IDCs, there were no statistically significant differences in the expression of the biomarkers in terms of either the percentage of cells staining or the immunoscores when comparing the entire tumor with each region except for the lower EGFR expression of arbitrarily selected region 1 and lower p53 expression of region 1 compared to that of the entire tumor section. For DCIS, there were no statistically significant differences in the expression of the biomarkers between the entire tumor and each region except in PCNA of region 2 compared to that of entire tumor section. Positive correlation of immunoscores was observed between the entire tumor and each region as well as across all four regions for IDC. Similar observations were noted with DCIS except for HER-2neu and PCNA. No statistically significant differences were observed in the absolute values of percent changes of biomarker expression between overall and the four regions for both DCIS and IDC. Therefore, no significant intratumor heterogeneity in the expression of HER-2neu, Bcl-2, and PCNA was observed in IDC. Minor regional variations were observed for EGFR and p53 in IDC. Similarly, no significant regional variation in the expression of markers was observed in DCIS except for PCNA.     

Polymerase chain reaction detection of Leishmania DNA in skin biopsy samples in Sri Lanka where the causative agent of cutaneous leishmaniasis is Leishmania donovani

Leishmania donovani is the known causative agent of both cutaneous (CL) and visceral leishmaniasis in Sri Lanka. CL is considered to be under-reported partly due to relatively poor sensitivity and specificity of microscopic diagnosis. We compared robustness of three previously described polymerase chain reaction (PCR) based methods to detectLeishmania DNA in 38 punch biopsy samples from patients presented with suspected lesions in 2010. Both, Leishmaniagenus-specific JW11/JW12 KDNA and LITSR/L5.8S internal transcribed spacer (ITS)1 PCR assays detected 92% (35/38) of the samples whereas a KDNA assay specific forL. donovani (LdF/LdR) detected only 71% (27/38) of samples. All positive samples showed a L. donovani banding pattern upon HaeIII ITS1 PCR-restriction fragment length polymorphism analysis. PCR assay specificity was evaluated in samples containing Mycobacterium tuberculosis, Mycobacterium leprae, and human DNA, and there was no cross-amplification in JW11/JW12 and LITSR/L5.8S PCR assays. The LdF/LdR PCR assay did not amplify M. leprae or human DNA although 500 bp and 700 bp bands were observed in M. tuberculosis samples. In conclusion, it was successfully shown in this study that it is possible to diagnose Sri Lankan CL with high accuracy, to genus and species identification, using Leishmania DNA PCR assays.