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Sacoglossa are known for stealing photosynthetically active chloroplasts from their macroalgal food and incorporating them into their cytosol. The nutritional support these alien organelles (kleptoplasts) provide to the slugs is still debatable. Comparing slugs starved in continuous darkness (non-photosynthetic condition) and light (photosynthetic condition) is often used to understand the contribution of the kleptoplasts to the slugs' metabolism. Here, we examined the slugs' side of starvation in darkness to better understand the effects of darkness on the slugs. We compared the gene expression profile and digestive activity of Elysia viridis, starved for one week under ambient light and continuous darkness. Starvation in darkness led to the up-regulation of genes related to glucose deficiency, while genes involved in the development, cellular organization, and reproduction were down-regulated. This specific gene expression may counteract reduced nutrient availability under non-photosynthetic conditions. Under photosynthetic conditions, kleptoplasts may have a higher nutritional value and may be able to support some metabolic processes. It appears that the slugs can only access kleptoplast photosynthates through autophagy during starvation. Nevertheless, autophagy and length reduction in darkness are highly elevated compared to light conditions, suggesting that more slug tissue is needed to satisfy the nutritional demands under non-photosynthetic conditions. Since we did not detect a gene expression related to the export of photosynthates to the slugs, our results support the hypothesis that slugs use kleptoplasts as larders accessible via autophagy. As long as the kleptoplasts are functional, they provide an energetic support, helping the slugs to reduce starvation-induced stress.

Aims Although few studies have examined the impact of surgical training on early postoperative outcomes in bariatric surgery, there is limited data on longer-term outcomes in trainee-performed cases. Our aim was to evaluate the effect of surgical training on weight loss outcomes following laparoscopic sleeve gastrectomy (LSG). Methods Data was prospectively collated on patients undergoing primary LSG at a Quaternary Bariatric London teaching Hospital between 2016-2017. Inclusion criteria was BMI≥35. Exclusion criteria were BMI 60, planned HDU admission and LSG with concomitant hiatus hernia repair. Operative time, length of stay, complications and longer-term excess weight loss was recorded with outcomes of consultant and trainee cases compared. Results 76 LSG patients were included; 44 performed by consultants, 32 by trainees. There was no difference in age, gender, pre-operative weight, BMI and number of obesity-related comorbidities between groups. Operative time (trainee105±10.0 vs consultant91±18.1 mins) and length of stay (trainee2.6±0.4 vs consultant2.8±0.9 days) were similar between groups. There were 3 complications in the trainee group (intra-abdominal collection requiring drainage, wound infection, hypokalaemia); and 2 with consultants (wound infection, intra-operative bleeding with ICU admission). Excess Weight Loss (%) at 2years was 55.9%±7.5% for trainee cases and 52.4%±6.7% for consultants(p = 0.49). Excess Weight Loss (%) at 3.5years was 54.9%±9.9% for trainee cases and 50.7%±9.9% for consultants(p = 0.54). Conclusion Outcomes in trainee-performed LSG are comparable to those performed by consultants. Surgical training in a high-volume teaching hospital does not appear to have detrimental effect on patient outcomes following LSG.

Aim Although few studies have examined the impact of surgical training on early postoperative outcomes in bariatric surgery, there is limited data on longer-term outcomes in trainee-performed cases. Our aim was to evaluate the effect of surgical training on weight loss outcomes following laparoscopic sleeve gastrectomy (LSG). Method Data was prospectively collated on patients undergoing primary LSG at a Quaternary Bariatric London teaching Hospital between 2016-2017. Inclusion criteria was BMI≥35. Exclusion criteria were BMI 60, planned HDU admission and LSG with concomitant hiatus hernia repair. Operative time, length of stay, complications and longer-term excess weight loss was recorded with outcomes of consultant and trainee cases compared. Results 76 LSG patients were included; 44 performed by consultants, 32 by trainees. There was no difference in age, gender, pre-operative weight, BMI and number of obesity-related comorbidities between groups. Operative time (trainee105±10.0 vs consultant91±18.1 mins) and length of stay (trainee2.6±0.4 vs consultant2.8±0.9 days) were similar between groups. There were 3 complications in the trainee group (intra-abdominal collection requiring drainage, wound infection, hypokalaemia); and 2 with consultants (wound infection, intra-operative bleeding with ICU admission). Excess Weight Loss(%) at 2 years was 55.9%±7.5% for trainee cases and 52.4%±6.7% for consultant cases(p=0.49). Excess Weight Loss(%) at 3.5 years was 54.9%±9.9% for trainee cases and 50.7%±9.9% for consultant cases(p=0.54). Conclusions Outcomes in trainee performed LSG are comparable to those performed by consultants. Surgical training in a high-volume teaching hospital does not appear to have detrimental effect on patient outcomes following LSG.

The emergence of novel variants of concern of SARS-CoV-2 demands a fast and reliable detection of such variants in local populations. Here we present a cost-efficient and fast workflow combining a pre-screening of SARS-CoV-2 positive samples using RT-PCR melting curve analysis with multiplexed IP-RP-HPLC-based single nucleotide primer extensions (SIRPH). The entire workflow from positive SARS-CoV-2 testing to base-specific identification of variants requires about 24 h. We applied the sensitive method to monitor the local VOC outbreaks in a few hundred positive samples collected in a confined region of Germany.

Dental caries is caused by acids released from bacterial biofilms. However, the in vivo formation of initial biofilms in relation to caries remains largely unexplored. The aim of this study was to compare the oral microbiome during the initial phase of bacterial colonization for individuals with (CC) and without (NC) cavitated dentin caries lesions. Bovine enamel slabs on acrylic splints were worn by the volunteers (CC: 14, NC: 13) for in situ biofilm formation (2 h, 4 h, 8 h, 1 ml saliva as reference). Sequencing of the V1/V2 regions of the 16S rRNA gene was performed (MiSeq). The relative abundances of individual operational taxonomic units (OTUs) were compared between samples from the CC group and the NC group. Random forests models were furthermore trained to separate the groups. While the overall heterogeneity did not differ substantially between CC and NC individuals, several individual OTUs were found to have significantly different relative abundances. For the 8 h samples, most of the significant OTUs showed higher relative abundances in the CC group, while the majority of significant OTUs in the saliva samples were more abundant in the NC group. Furthermore, using OTU signatures enabled a separation between both groups, with area-under-the-curve (AUC) values of ~0.8. In summary, the results suggest that initial oral biofilms provide the potential to differentiate between CC and NC individuals.

Requirements Engineering is a multidisciplinary and a human-centered process, therefore, the artifacts produced from RE are always error-prone. The most significant of these errors are missing or misunderstanding requirements. Information loss in RE could result in omitted logic in the software, which will be onerous to correct at the later stages of development. In this paper, we demonstrate and investigate how interactive and Loud Paper Prototyping (LPP) can be integrated to collect stakeholders’ needs and expectations than interactive prototyping or face-to-face meetings alone. To this end, we conducted a case study of (1) 31 mobile application (App) development teams who applied either of interactive or loud prototyping and (2) 19 mobile App development teams who applied only the face-to-face meetings. From this study, we found that while using Silent Paper Prototyping (SPP) rather than No Paper Prototyping (NPP) is a more efficient technique to capture Non-Functional Requirements (NFRs), User Interface (UI) requirements, and existing requirements, LPP is more applicable to manage NFRs, UI requirements, as well as adding new requirements and removing/modifying the existing requirements. We also found that among LPP and SPP, LPP is more efficient to capture and influence Functional Requirements (FRs).

Exit from quiescence and reentry into cell cycle is essential for HSC self-renewal and regeneration. Skp2 is the F-box unit of the SCF E3-ligase that targets the CDK inhibitors (CKIs) p21Cip1, p27Kip1, p57Kip2, and p130 for degradation. These CKIs inhibit the G1 to S-phase transition of the cell cycle, and their deletion results in increased cell proliferation and decreased stem cell self-renewal. Skp2 deletion leads to CKIs stabilization inducing cell-cycle delay or arrest, and conversely, increased Skp2 expression is often found in cancers. Here, we show that SKP2 expression is increased in HSC and progenitors in response to hematopoietic stress from myelosuppression or after transplantation. At steady state, SKP2 deletion decreased the mitotic activity of HSC and progenitors resulting in enhanced HSC quiescence, increased HSC pool size, and maintenance. However, the inability to rapidly enter cell cycle greatly impaired the short-term repopulating potential of SKP2 null HSC and their ability to regenerate after myeloablative stress. Mechanistically, deletion of SKP2 in HSC and progenitors stabilized CKIs in vivo, particularly p27Kip1, p57Kip2, and p130. Our results demonstrate a previously unrecognized role for SKP2 in regulating HSC and progenitor expansion and hematopoietic regeneration after stress.

Macrophages are a major cellular constituent of the tumour stroma and contribute to breast cancer prognosis. The precise role and treatment strategies to target macrophages remain elusive. As macrophage infiltration is associated with poor prognosis and high grade tumours we used the THP-1 cell line to model monocyte-macrophage differentiation in co-culture with four breast cancer cell lines (MCF7, T47D, MDA-MB-231, MDA-MB-468) to model in vivo cellular interactions. Polarisation into M1 and M2 subtypes was confirmed by specific cell marker expression of ROS and HLA-DR, respectively. Co-culture with all types of macrophage increased migration of ER-positive breast cancer cell lines, while M2-macrophages increased mammosphere formation, compared to M1-macrophages, in all breast cancer cells lines. Treatment of cells with Zoledronate in co-culture reduced the "pro-tumourigenic" effects (increased mammospheres/migration) exerted by macrophages. Direct treatment of breast cancer cells in homotypic culture was unable to reduce migration or mammosphere formation.Macrophages promote "pro-tumourigenic" cellular characteristics of breast cancer cell migration and stem cell activity. Zoledronate targets macrophages within the microenvironment which in turn, reduces the "pro-tumourigenic" characteristics of breast cancer cells. Zoledronate offers an exciting new treatment strategy for both primary and metastatic breast cancer.

Hypercalciuria is a frequent cause of non-glomerular hematuria in pediatric patients. Because hypercalciuria can be secondary to primary hyperparathyroidism, measurement of serum parathyroid hormone (PTH) levels is often performed in children with this urinary abnormality. A retrospective chart review was performed to determine the diagnostic yield of PTH measurements when performed under these clinical circumstances. Over a 30-month period (January 1, 2001 to September 30, 2003), among 31 children who had a PTH determination, the level was elevated in 1 (3%) patient. Based on these findings and the serious nature of untreated primary hyperparathyroidism, serum PTH level should be measured in pediatric patients with newly diagnosed hypercalciuria.

Background: Stromal-epithelial interaction is a key factor in tumour progression. Cancer-associated fibroblasts (CAFs) and macrophage infiltration have been associated with early relapse in breast cancer. Bisphosphonates are effective inhibitors of osteoclast activation in metastatic breast cancer but also have a general inhibitory effect on breast cancer progression. In order to monitor a potential tumour stromal response in breast cancer during treatment with an aromatase inhibitor and a bisphosphonate we analysed pre-and post-treatment samples from a neoadjuvant window study and focused on the presence of macrophages and CAFs. Materials and methods: Tissue microarrays (TMAs) from surgical samples and pre-operative core biopsies were immunohistochemically stained for aSMA (CAF marker), CD68 (macrophages) and epithelial proliferation (Ki67). In order to validate if the presence of macrophages and aSMA could be monitored by the TMA approach, we initially analysed a screening cohort of 144 breast cancer samples. We then studied pre-and post-treatment samples from 110 postmenopausal ER-positive invasive breast cancer patients randomised to receive 14 days of preoperative treatment (placebo, Letrozole, or Letrozole plus Zoledronate). Results: In the screening cohort, we observed significant links between aSMA positive fibroblasts and disease recurrence as well as between CD68 positive macrophages and tumour size, grade, lymph node positivity and recurrence. This validated the use of TMAs for stromal analyses and furthersupported a link with key tumour biological events. In both treatment arms, there was a significant drop in absolute Ki67 value compared to placebo (-9.3% Letrozole and -13.1% combination reduction versus 1% increase, P Conclusion: Short term treatment response in the epithelial component of cancers was paralleled by specific responses in the tumour stromal component. These novel findings suggest that bisphosphonates and aromatase inhibitors have major effects on tumour stroma in vivo which might augment their inhibitory effect on tumour progression. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P4-05-05.