Biopsy Sampling in Upper Gastrointestinal Endoscopy: A Survey from 10 Tertiary Referral Centres Across Europe.

BACKGROUND: Guidelines give robust recommendations on which biopsies should be taken when there is endoscopic suggestion of gastric inflammation. Adherence to these guidelines often seems arbitrary. This study aimed to give an overview on current practice in tertiary referral centres across Europe. METHODS: Data were collected at 10 tertiary referral centres. Demographic data, the indication for each procedure, endoscopic findings, and the number and sampling site of biopsies were recorded. Findings were compared between centres, and factors influencing the decision to take biopsies were explored. RESULTS: Biopsies were taken in 56.6% of 9,425 procedures, with significant variation between centres (p < 0.001). Gastric biopsies were taken in 43.8% of all procedures. Sampling location varied with the procedure indication (p < 0.001) without consistent pattern across the centres. Fewer biopsies were taken in centres which routinely applied the updated Sydney classification for gastritis assessment (46.0%), compared to centres where this was done only upon request (75.3%, p < 0.001). This was the same for centres stratifying patients according to the OLGA system (51.8 vs. 73.0%, p < 0.001). More biopsies were taken in centres following the MAPS guidelines on stomach surveillance (68.1 vs. 37.1%, p < 0.001). Biopsy sampling was more likely in younger patients in 8 centres (p < 0.05), but this was not true for the whole cohort (p = 0.537). The percentage of procedures with biopsies correlated directly with additional costs charged in case of biopsies (r = 0.709, p = 0.022). CONCLUSION: Adherence to guideline recommendations for biopsy sampling at gastroscopy was inconsistent across the participating centres. Our data suggest that centre-specific policies are applied instead.

Variation in outer blubber lipid concentration does not reflect morphological body condition in humpback whales.

An animal's body condition provides valuable information for ecophysiological studies, and is an important measure of fitness in population monitoring and conservation. While both the external body shape of an animal and its internal tissues (i.e. fat content) can be used as a measure of body condition, the relationship between the two is not always linear. We compared the morphological body condition (external metric obtained through aerial photogrammetry) of migrating humpback whales (Megaptera novaeangliae) with their outer blubber lipid concentration (internal metric obtained through blubber biopsy sampling) off the coast of south-west Australia early and late in the breeding season (spanning ∼4.5 months). The external body condition index of juvenile and adult humpback whales decreased by 26.9 (from 18.8% to -8.1%) and 12.0 percentage points (from 8.6% to -3.4%), respectively, between the early and late phase. In contrast, we found no intra-seasonal change in blubber lipid concentration, and no difference between reproductive classes (juveniles, adults and lactating females); however, the small sample size prevented us from effectively testing these effects. Importantly, however, in the 33 animals for which paired metrics were obtained, we found no correlation between the morphometric body condition index and the blubber lipid concentration of individual whales. The lack of a linear relationship suggests that changes in outer blubber lipid concentration do not reflect external changes in body shape, thus limiting the utility of outer blubber lipid reserves for individual body condition evaluation. The wider spectrum of change in body morphometry captured with aerial photogrammetry supports the use of body morphometry as a reliable and well-tested method.

Virtual liver needle biopsy from reconstructed three-dimensional histopathological images: Quantification of sampling error.

INTRODUCTION: Prevalently considered as the "gold-standard" for diagnosis of hepatic fibrosis and cirrhosis, the clinical liver needle biopsy is known to be subject to inadequate sampling and a high mis-sampling rate. However, quantifying such sampling bias has been difficult as generating a large number of needle biopsies from the same living patient is practically infeasible. We construct a three-dimension (3D) virtual liver tissue volume by spatially registered high resolution Whole Slide Images (WSIs) of serial liver tissue sections with a novel dynamic registration method. We further develop a Virtual Needle Biopsy Sampling (VNBS) method that mimics the needle biopsy sampling process. We apply the VNBS method to the reconstructed digital liver volume at different tissue locations and angles. Additionally, we quantify Collagen Proportionate Area (CPA) in all resulting virtual needle biopsies in 2D and 3D. RESULTS: The staging score of the center 2D longitudinal image plane from each 3D biopsy is used as the biopsy staging score, and the highest staging score of all sampled needle biopsies is the diagnostic staging score. The Mean Absolute Difference (MAD) in reference to the Scheuer and Ishak diagnostic staging scores are 0.22 and 1.00, respectively. The absolute Scheuer staging score difference in 22.22% of sampled biopsies is 1. By the Ishak staging method, 55.56% and 22.22% of sampled biopsies present score difference 1 and 2, respectively. There are 4 (Scheuer) and 6 (Ishak) out of 18 3D virtual needle biopsies with intra-needle variations. Additionally, we find a positive correlation between CPA and fibrosis stages by Scheuer but not Ishak method. Overall, CPA measures suffer large intra- and inter- needle variations. CONCLUSIONS: The developed virtual liver needle biopsy sampling pipeline provides a computational avenue for investigating needle biopsy sampling bias with 3D virtual tissue volumes. This method can be applied to other tissue-based disease diagnoses where the needle biopsy sampling bias substantially affects the diagnostic results.

Complex Internal Microstructure of Feather Follicles on Chicken Skin Promotes the Bacterial Cross-Contamination of Carcasses During the Slaughtering Process.

Chicken skin is considered the most susceptible to bacterial contamination during slaughter. It is rich in bushy feather follicles with complex internal structures that can absorb bacteria via cross-contamination during slaughter. Until now, the microstructural changes and local bacterial composition of feather follicles during slaughter have not been thoroughly investigated. This study used hematoxylin-eosin (HE) staining of the tissue paraffin section to investigate the structure of the feather follicles on chicken skin. In addition, the biopsy sampling method was employed for the high-throughput sequencing of 16S RNA genes to study the composition and source of bacterial contamination during slaughter. The results show that the feather follicles on chicken skin form a closed cavity structure during the slaughtering process. The volume of the irregular follicle cavity was about Ø: 200 µm × D: 1040 µm, which provides a place for the bacteria to absorb and resist the cleaning and disinfection during the slaughtering process. The composition of bacteria in the feather follicle was mainly Acinetobacter (37%), Psychrobacter (8%), Macrococcus (5%), and Comamonas (2%). The heat map obtained via the species abundance analysis of the feather follicle samples as well as the slaughter environment samples suggests that the gastrointestinal feces contaminated the feather follicles on the chicken skin mainly during the evisceration, defeathering, and chilling processes, and the last-stage chilling water also caused severe cross-contamination to the feather follicles during the chilling process.

The impact of biopsy sampling errors and the quality of surgical margins on local recurrence and survival in chondrosarcoma.

PURPOSE: To examine the frequency of computed tomography (CT)-guided biopsy sampling errors in chondrosarcomas, as well as the impact of these errors and the achieved surgical margins on local recurrence-free survival (LRFS) and disease-specific survival (DSS). MATERIAL AND METHODS: A total of 68 consecutive patients treated for chondrosarcoma from 2000-2015 were retrospectively reviewed with a minimum follow-up duration of 2 years. RESULTS: The primary location was at the extremities in 46 patients (67.6%) and at the axial skeleton in 22 patients (32.4%). Seven patients underwent planned intralesional curettage. Surgical margins were assessed in the remaining 53 patients and included 21 wide (39.6%), 25 marginal (47.1%), and seven intralesional (13.2%) resections. Biopsy sampling errors occurred in ten patients (14.7%). LRFS was 82.2±7.8% at 5 years and 76.9±7.8% at 10 years. An intact anatomical barrier was associated with the most preferable LRFS of 89±10.5% after 10 years. DSS was 79.2±8.5% at 5 years and 75.5±6.4% at 10 years. The metric distance of the surgical margin and the presence of a biopsy sampling error did not affect either LRFS or DSS. CONCLUSION: Even though histological grading in chondrosarcoma is difficult, sampling errors in preoperative biopsies are relatively rare and do not adversely affect outcomes. The presence of an anatomical barrier has a greater impact on LRFS than the metric distance of the surgical margins.

Genetic divergence between two phenotypically distinct bottlenose dolphin ecotypes suggests separate evolutionary trajectories.

Due to their worldwide distribution and occupancy of different types of environments, bottlenose dolphins display considerable morphological variation. Despite limited understanding about the taxonomic identity of such forms and connectivity among them at global scale, coastal (or inshore) and offshore (or oceanic) ecotypes have been widely recognized in several ocean regions. In the Southwest Atlantic Ocean (SWA), however, there are scarce records of bottlenose dolphins differing in external morphology according to habitat preferences that resemble the coastal-offshore pattern observed elsewhere. The main aim of this study was to analyze the genetic variability, and test for population structure between coastal (n = 127) and offshore (n = 45) bottlenose dolphins sampled in the SWA to assess whether their external morphological distinction is consistent with genetic differentiation. We used a combination of mtDNA control region sequences and microsatellite genotypes to infer population structure and levels of genetic diversity. Our results from both molecular marker types were congruent and revealed strong levels of structuring (microsatellites FST = 0.385, p < .001; mtDNA FST =  0.183, p < .001; ΦST = 0.385, p < .001) and much lower genetic diversity in the coastal than the offshore ecotype, supporting patterns found in previous studies elsewhere. Despite the opportunity for gene flow in potential "contact zones", we found minimal current and historical connectivity between ecotypes, suggesting they are following discrete evolutionary trajectories. Based on our molecular findings, which seem to be consistent with morphological differentiations recently described for bottlenose dolphins in our study area, we recommend recognizing the offshore bottlenose dolphin ecotype as an additional Evolutionarily Significant Unit (ESU) in the SWA. Implications of these results for the conservation of bottlenose dolphins in SWA are also discussed.

A randomized trial of local endometrial injury during ovulation induction cycles.

OBJECTIVE: To investigate the effect of endometrial injury on pregnancy outcomes among infertile women taking clomifene citrate. METHODS: A prospective randomized trial was undertaken at an Egyptian hospital between January 26, 2015, and July 17, 2016. Eligible women (≥1 year primary/secondary/unexplained infertility, aged 20-35 years, day-2 follicle-stimulating hormone <12 IU/L, normal prolactin/thyroid function/uterine cavity, ≥1 patent tube, male partner with normal semen count and motility, 3 cycles of clomifene citrate without success) were randomly allocated (1:1) using computer-generated numbers into intervention (received endometrial injury on cycle day 15-24) or control groups (sham procedure). Women began ovulation induction on days 3-5 of the following cycle. Participants and investigators were not masked to group assignment. The primary outcomes were clinical pregnancy, spontaneous abortion, and multiple pregnancy rates. Women who completed follow-up were included in analyses. RESULTS: The intervention group included 52 women and the control group 53 women. The clinical pregnancy rate was significantly higher in the intervention group (37% [n=19]) than in the control group (13% [n=7]; P=0.006). No differences between the intervention and control groups were noted for spontaneous abortion rate (11% [2/19] vs 29% [2/7]; P=0.287) or multiple pregnancy rate (11% [2/19] vs 14% [1/7]; P=0.790). No adverse effects were reported. CONCLUSION: Endometrial injury before ovulation induction could improve chances of pregnancy and its outcomes. CLINICALTRIALS.GOV: NCT02345837.

Negative HPV testing among patients with biopsy-proven cervical intraepithelial neoplasia grade 2/3 or cervical cancer.

OBJECTIVE: To determine the proportion of patients diagnosed with cervical cancer or cervical intraepithelial neoplasia grade 2 or 3 (CIN 2/3) despite testing negative for HPV. METHODS: A prospective study was conducted at a university hospital in Barcelona, Spain, between March 2003 and January 2015 among women with abnormal cytology results according to the Bethesda classification system. All participants underwent HPV testing, cytology, and colposcopy. RESULTS: Among 1376 participants, 609 (44.3%) were diagnosed with CIN 2/3. Of these women, 74 (12.2%) tested negative for HPV. Among 479 women with colposcopy-guided biopsy sampling showing CIN 2/3, cone biopsy results that were negative for CIN 2/3 were more frequent among patients with negative HPV testing (13/61 [21.3%]) than among those with positive HPV tests (58/418 [13.9%]; P=0.03). Additionally, among 59 patients diagnosed with cervical cancer, 9 (15.3%) tested negative for HPV. CONCLUSION: The HPV test was negative for 12% and 15% of patients diagnosed with biopsy-proven CIN 2/3 and cervical cancer, respectively.

Threshold for endometrial sampling among postmenopausal patients without vaginal bleeding.

OBJECTIVE: To provide an optimum threshold for endometrial biopsy sampling among postmenopausal women without vaginal bleeding and an incidentally-found endometrial lining of above 4mm. METHODS: A cohort of postmenopausal women (aged ≥50 years) who underwent pelvic ultrasonography at a tertiary US hospital for indications other than vaginal bleeding was retrospectively evaluated. Women were included if they had an endometrial lining of above 4mm. Logistic regression was performed to determine the probability of endometrial carcinoma and atypical hyperplasia at each increasing millimeter of endometrial thickness from 4 to 20mm. RESULTS: Among 462 women, carcinoma was identified in 9 (1.9%) and atypical hyperplasia in 7 (1.5%). An endometrial thickness of or above 14 mm was significantly associated with atypical hyperplasia (odds ratio 4.29; 95% confidence interval 1.30-14.20; P=0.02), with a negative predictive value of 98.3%. A thickness of or above 15 mm was associated with carcinoma (odds ratio 4.53; 95% confidence interval 1.20-17.20; P=0.03), with a negative predictive value of 98.5% and a 0.06% risk of cancer. CONCLUSION: Irrespective of conventional risk factors, an incidentally-found thickened endometrial lining of less than 15 mm might not warrant endometrial biopsy sampling among postmenopausal women without vaginal bleeding.

The distribution and stratification of persistent organic pollutants and fatty acids in bottlenose dolphin (Tursiops truncatus) blubber.

Blubber has been used for decades to monitor exposure of marine mammals to persistent organic pollutants (POPs). However, little is known about POP variability as a function of blubber depth and across the body of the animal. Remote blubber biopsy sampling (e.g, projectile biopsy) is the most common technique used to acquire samples from free-swimming animals, yet such techniques may result in variable sampling. It is important to understand whether blubber stratification or body location affects POP concentration or the concentration of other important blubber constituents such as fatty acids (FA). To investigate the influence of sampling depth and location on POP concentration, full depth blubber samples were taken from one stranded bottlenose dolphin (Tursiops truncatus) at six different body sites to assess variation in FA distribution and contaminant storage with body location. Three of the samples from different body locations were separated into histologically distinct layers to examine the effect of blubber depth and body location on POPs and FAs. In this individual, both POPs and FAs were heterogeneous with blubber depth and body location. POP concentrations were significantly greater in ventral (average ΣPBDEs 1350 ng/g lipid) and anterior (average ΣPCBs 28,700 ng/g lipid) body locations and greater in the superficial blubber layer (average ΣPCBs 35,500 ng/g lipid) when compared to the deep (8390 ng/g lipid) and middle (23,700 ng/g lipid) layers. Proportionally more dietary FAs were found in dorsal blubber and in middle and deep layers relative to other locations while the reverse was true for biosynthesized FAs. Stratification was further examined in blubber from the same body location in five additional stranded bottlenose dolphins. Although FAs were stratified with blubber depth, lipid-normalized POPs were not significantly different with depth, indicating that POP concentrations can vary in an individual with blubber depth though the direction of POP stratification is not consistent among individuals.