Antibiotic-exposed patients with non-small-cell lung cancer preserve efficacy outcomes following first-line chemo-immunotherapy.

BACKGROUND: Prior antibiotic therapy (pATB) is known to impair efficacy of single-agent immune checkpoint inhibitors (ICIs), potentially through the induction of gut dysbiosis. Whether ATB also affects outcomes to chemo-immunotherapy combinations is still unknown. PATIENTS AND METHODS: In this international multicentre study, we evaluated the association between pATB, concurrent ATB (cATB) and overall survival (OS), progression-free survival (PFS) and objective response rate (ORR) in patients with non-small-cell lung cancer (NSCLC) treated with first-line chemo-immunotherapy at eight referral institutions. RESULTS: Among 302 patients with stage IV NSCLC, 216 (71.5%) and 61 (20.2%) patients were former and current smokers, respectively. Programmed death-ligand 1 tumour expression in assessable patients (274, 90.7%) was ≥50% in 76 (25.2%), 1%-49% in 84 (27.9%) and <1% in 113 (37.5%). Multivariable analysis showed pATB-exposed patients to have similar OS {hazard ratio (HR) = 1.42 [95% confidence interval (CI): 0.91-2.22]; P = 0.1207} and PFS [HR = 1.12 (95% CI: 0.76-1.63); P = 0.5552], compared to unexposed patients, regardless of performance status. Similarly, no difference with respect to ORR was found across pATB exposure groups (42.6% versus 57.4%, P = 0.1794). No differential effect was found depending on pATB exposure duration (≥7 versus <7 days) and route of administration (intravenous versus oral). Similarly, cATB was not associated with OS [HR = 1.29 (95% CI: 0.91-1.84); P = 0.149] and PFS [HR = 1.20 (95% CI: 0.89-1.63); P = 0.222] when evaluated as time-varying covariate in multivariable analysis. CONCLUSIONS: In contrast to what has been reported in patients receiving single-agent ICIs, pATB does not impair clinical outcomes to first-line chemo-immunotherapy of patients with NSCLC. pATB status should integrate currently available clinico-pathologic factors for guiding first-line treatment decisions, whilst there should be no concern in offering cATB during chemo-immunotherapy when needed.

Obstetric intra-operative cell salvage: a review of an established cell salvage service with 1170 re-infused cases.

The use of cell salvage during caesarean section has been increasing steadily, although there are concerns relating to cost, a perceived risk of amniotic fluid embolism, and fetal red cell sensitisation. We present observational data from almost a decade of use of intra-operative cell salvage in obstetrics. By the end of this period, we set up cell salvage collection for > 98% of all caesarean sections. From 2008 to 2017, 1170 women have had a re-infusion of cell salvaged blood with no clinical safety concerns; the median (IQR [range]) volume was 231 (154-306 [80-1690]) ml. During this time there has been a marked reduction in the number of women who were transfused allogeneic blood, as well as the amount of blood transfused. In total, 647 (55%) women have had alloimmunisation testing, with two positive cases. Quality control data indicate that the quality of blood processed from partial first bowls is no worse than that from full bowls. We discuss the costs of providing this service with regard to: staffing costs; single suction; leucodepletion filters; selectivity in the processing of collected blood; and the use of partial first bowls.

Obstetric intra-operative cell salvage and maternal fetal red cell contamination.

BACKGROUND: The significance of fetal red blood cell (RBC) contamination in obstetric intra-operative cell salvage is not fully known. It is unclear if we re-infuse a larger volume of fetal RBCs into the maternal circulation than the amount that occurs secondary to transplacental haemorrhages is unclear. We also do not know if there is a critical volume required to cause alloimmunisation or if larger volumes increase the risk. OBJECTIVES: The aim of this study is to provide data on the level of fetal RBC contamination in the maternal circulation prior to delivery and immediately post-partum and to compare these levels to those found in processed cell-salvaged blood. METHODS: In the first part of this study, we quantified the levels of fetal RBCs circulating in women immediately prior to delivery. This was then repeated with a separate group measuring the levels of fetal RBCs pre- and post-delivery. RESULTS: We found that 37% of women had fetal cells detected in their circulation, median 0·00 mL (IQR 0-0·24; average 0·3 mL, maximum 4·56 mL). Fetal RBCs were present pre-delivery (maximum 0·66 mL) in 16% of women, increasing to 53% post-delivery (median 0·66 mL; IQR 0·22-2·20, maximum 21·20 mL). CONCLUSIONS: We have shown that fetal RBCs are present in the maternal circulation throughout pregnancy and that the volumes are comparable to that obtained from intra-operative salvage, with contamination amounts of up to 19 mL. At the Royal Cornwall Hospital, our experience and evidence supports offering intra-operative salvage to all women, and we have not noted an increase in antibody formation, compared to allogeneic transfusion.

A Study of the Patients Admitted to the Clinical Decision Unit over One Year.

Clinical decision units (CDUs) are areas within an emergency department (ED) providing care for the patient who may benefit from an extended observation period, usually for a maximum of twenty-four hours. A retrospective patient record audit was performed to determine the characteristics of patients admitted to the Cork University Hospital (CUH) CDU over 12 months. The average length of stay of a patient in the CDU was 29 hours. The most common diagnoses admitted to the CDU were chest pain (9.5%) and headache (7.2%). The research implies that the CDU provided a means for CUH to save approximately €2 million annually.

EGFR ligands and DNA repair genes: genomic predictors of complete response after capecitabine-based chemoradiotherapy in locally advanced rectal cancer.

Epidermal growth factor receptor (EGFR) activation by radiation leads to increased cell proliferation and acts as a radioresistance mechanism. Neoadjuvant chemoradiation is the standard of care for locally advanced rectal cancer, and to date, no biomarkers of response have been found. We analyzed polymorphisms in the EGFR and its ligands, DNA repair genes and the thymidylate synthase in 84 stages II and III rectal cancer patients treated with neoadjuvant capecitabine plus radiotherapy. The rs11942466 polymorphism in the amphiregulin (AREG) gene region was associated with a pathological complete response (ypCR) (odds ratio: 0.26; 95% confidence interval: 0.06-0.79; P=0.014). The rs11615 C>T polymorphism in the ERCC1 gene also correlated with the ypCR as no patients with a C/C genotype achieved ypCR; P=0.023. This is the first work to propose variants within the AREG and the ERCC1 genes as promising predictive biomarkers of ypCR in rectal cancer.

Chemotactic clusters in confined run-and-tumble bacteria: a numerical investigation.

We present a simulation study of pattern formation in an ensemble of chemotactic run-and-tumble bacteria, focussing on the effect of spatial confinement, either within traps or inside a maze. These geometries are inspired by previous experiments probing pattern formation in chemotactic strains of E. coli under these conditions. Our main result is that a microscopic model of chemotactic run-and-tumble particles which themselves secrete a chemoattractant is able to reproduce the main experimental observations, namely the formation of bacterial aggregates within traps and in dead ends of a maze. Our simulations also demonstrate that stochasticity plays a key role and leads to a hysteretic response when the chemotactic sensitivity is varied. We compare the results of run-and-tumble particles with simulations performed with a simplified version of the model where the active particles are smooth swimmers which respond to chemotactic gradients by rotating towards the source of chemoattractant. This class of models leads again to aggregation, but with quantitative and qualitative differences in, for instance, the size and shape of clusters.

Assessment of intra-operative cell salvage haemolysis in the obstetric and orthopaedic clinical setting, in comparison with allogeneic blood.

OBJECTIVES/BACKGROUND: Haemolysis is still a re-occurring theme in intra-operative cell salvage (ICS) with further haemolysis possibly caused by suction pressure, washing/centrifuging process and aspiration method. Previous investigations, along with manufacturer's reports, state that between 75 and 95% of free haemoglobin (Hb) is removed by the washing and centrifugation process; however, if these results are above the expected levels, excess free Hb may remain after washing. The aim of this article was to quantify haemolysis levels whilst employing different aspiration methods from skimmed (orthopaedics) and pooled (obstetrics) surgery types and comparing this to allogeneic blood. METHODS/MATERIALS: Samples obtained from 50 allogeneic units and 50 ICS cases (25 obstetric and 25 orthopaedic) were tested for plasma free Hb levels. RESULTS: Free Hb testing as a marker of haemolysis was greatest in orthopaedic 17·2 g L(-1) (range: 1·7-57·0 g L(-1) ), obstetric 2·8 g L(-1) (range: 1·0-13·5 g L(-1) ) and allogeneic 0·95 g L(-1) (range: 0·2-4·8 g L(-1) ) cases. CONCLUSION: ICS involving skimming collection techniques (orthopaedics) had significantly more haemolysis than pooled collections (obstetrics) (P < 0·001). Further analysis of orthopaedic data highlighted a difference between the three machines used with the Haemonetics OrthoPat (Haemonetics Ltd., Watford, UK) significantly higher with a free Hb of 29·8 g L(-1) compared with the other two machines 6·7 g L(-1) (P < 0·001). On comparison of ICS blood to allogeneic blood, free Hb levels obtained from ICS were significantly higher (P < 0·001).

Lactate dehydrogenase and Haemolysis Index as quality control markers of haemolysis in intra-operative cell salvage.

OBJECTIVES: The aim of this investigation was to explore the potential use of the tests lactate dehydrogenase (LDH) and Haemolysis Index as haemolysis markers in intra-operative cell salvage (ICS) blood in comparison to plasma free haemoglobin levels. BACKGROUND: Quality control (QC) should be seen as a fundamental part of any ICS blood conservation programme, however, due to lack of available knowledge, familiarity and experience, QC is still a comparatively new subject. A QC pilot scheme is currently being undertaken by the Royal Cornwall Hospital in association with the UK Cell Salvage Action Group to explore potential markers that can be used to assess the quality of blood obtained from ICS. This test list should be available to all ICS users and achievable within financial budgets. Currently this proposed test list includes a full blood count, a protein marker such as urine albumin/microalbumin and heparin monitoring. Haemolysis testing is another key marker. METHODS/MATERIALS: Samples were collected from ICS processed blood and allogeneic SAGM leucodepleted red cell units and processed for plasma free haemoglobin, LDH and Haemolysis Index. RESULTS: There was a very strong correlation between plasma free haemoglobin and LDH (0.960), and plasma free haemoglobin and the Haemolysis Index (0.944). CONCLUSION: We have shown that the LDH and Haemolysis Index tests are suitable and reliable alternatives for measuring haemolysis from samples obtained from ICS or allogeneic blood. We have incorporated the LDH test into our Hospital's ICS QC package and recommend that this test is considered for all ICS QC samples.

Exposure to the common food additive carrageenan leads to glucose intolerance, insulin resistance and inhibition of insulin signalling in HepG2 cells and C57BL/6J mice.

AIMS/HYPOTHESIS: The aim of this study was to determine the impact of the common food additive carrageenan (E407) on glucose tolerance, insulin sensitivity and insulin signalling in a mouse model and human hepatic cells, since carrageenan is known to cause inflammation through interaction with toll-like receptor (TLR)4, which is associated with inflammation in diabetes. METHODS: Male C57BL/6J mice were given carrageenan (10 mg/l) in their drinking water, and underwent a glucose tolerance test (GTT), an insulin tolerance test (ITT) and an ante-mortem intraperitoneal insulin injection. HepG2 cells were exposed to carrageenan (1 mg/l × 24 h) and insulin. Levels of phospho(Ser473)-protein kinase B (Akt), phospho(Ser307)-IRS1, phosphoinositide 3-kinase (PI3K) activity and phospho(Ser32)-inhibitor of κB (IκBα) were determined by western blotting and ELISA. RESULTS: Glucose tolerance was significantly impaired in carrageenan-treated 12-week-old mice compared with untreated controls at all time points (n = 12; p < 0.0001). Baseline insulin and insulin levels at 30 min after taking glucose during the GTT were significantly higher following carrageenan treatment. During the ITT, glucose levels declined by more than 80% in controls, but not in carrageenan-treated mice. Carrageenan exposure completely inhibited insulin-induced increases in phospho-(Ser473)-Akt and PI3K activity in vivo in mouse liver and in human HepG2 cells. Carrageenan increased phospho(Ser307)-IRS1 levels, and this was blocked when carrageenan-induced inflammation was inhibited. CONCLUSION: This is the first report of the impact of carrageenan on glucose tolerance and indicates that carrageenan impairs glucose tolerance, increases insulin resistance and inhibits insulin signalling in vivo in mouse liver and human HepG2 cells. These effects may result from carrageenan-induced inflammation. The results demonstrate extra-colonic manifestations of ingested carrageenan and suggest that carrageenan in the human diet may contribute to the development of diabetes.

A modified thrombin clotting time test as a quality control marker for heparin contamination in obstetric intraoperative cell salvage.

OBJECTIVE: To assess if a modified thrombin clotting time test could be used as a simple quality control (QC) method to screen for unfractionated heparin in the product obtained from obstetric intraoperative cell salvage cases before re-infusion. BACKGROUND: A national QC scheme has recently been piloted to monitor the quality of autologous blood being returned to the patient. Laboratory tests include full blood count and microalbumin. Unfractionated heparin testing should be performed to ensure that there is no gross contamination of heparin in the final product; however, presently, there is no quick cheap test available suitable for heparin detection. MATERIALS AND METHODS: Samples were collected into plain non-anticoagulated tubes and centrifuged at 2500 × g for 5 min. Supernatant was mixed with commercially available coagulated normal plasma and a thrombin clotting time test performed. RESULTS: Calibration runs demonstrated that our system was sensitive up to 0 · 14 IU mL(-1) heparin, linear between 0 · 08 and 0 · 14 IU mL(-1). CONCLUSION: We have shown that the thrombin clotting time test can be modified and used as a cheap and reliable marker for heparin contamination. We have successfully incorporated this modified test into our hospital's obstetric QC scheme.

SEOM-GECP-GETTHI Clinical Guidelines for the treatment of patients with thymic epithelial tumours (2021).

Thymic epithelial tumours (TET) represent a heterogeneous group of rare malignancies that include thymomas and thymic carcinoma. Treatment of TET is based on the resectability of the tumour. If this is considered achievable upfront, surgical resection is the cornerstone of treatment. Platinum-based chemotherapy is the standard regimen for advanced TET. Due to the rarity of this disease, treatment decisions should be discussed in specific multidisciplinary tumour boards, and there are few prospective clinical studies with new strategies. However, several pathways involved in TET have been explored as potential targets for new therapies in previously treated patients, such as multi-tyrosine kinase inhibitors with antiangiogenic properties and immune checkpoint inhibitors (ICI). One third of patient with thymoma present an autoimmune disorders, increasing the risk of immune-related adverse events and autoimmune flares under ICIs. In these guidelines, we summarize the current evidence for the therapeutic approach in patients with TET and define levels of evidence for these decisions.

Intraoperative cell salvaged blood as part of a blood conservation strategy in Caesarean section: is fetal red cell contamination important?

BACKGROUND: Cell salvage is used in obstetric surgery as part of a blood conservation strategy in our Trust. This carries a theoretical risk of amniotic fluid embolism and also a risk of fetal red cells being present in the re-infusion, resulting in alloimmunization. In this study, we attempted to quantify the risk of antibody formation from re-infusion of autologous blood after Caesarean section. METHODS: Women presenting for elective Caesarean section were routinely requested to consent for collection of blood by cell salvage, using one suction device. If an adequate volume of blood was collected, it was processed and, if clinically appropriate, re-infused via a leucodepletion filter. Women who received a re-infusion were followed up to test for antibody formation. RESULTS: Seventy women consented for re-infusion and follow-up. The median volume re-infused was 324 ml (range 118-1690 ml). The median fetal red cell contamination was 0.8 ml (range 0.2-12.9 ml). All re-infusions were given without adverse clinical signs. No antibodies were detected in 48 follow-up samples. One positive anti-S antibody was detected. CONCLUSIONS: The implementation of a blood conservation strategy which includes the use of intraoperative cell salvage appears safe and can contribute to a reduction in the number of blood transfusions to the obstetric population. We remain uncertain of the significance of fetal red cell contamination.

Targeting HER2 in non-small-cell lung cancer (NSCLC): a glimpse of hope? An updated review on therapeutic strategies in NSCLC harbouring HER2 alterations.

Non-small-cell lung cancer (NSCLC) harbouring HER2 alterations is now considered a distinct molecular subtype. The activation of HER2 in NSCLC occurs via three mechanisms, i.e. gene mutation (1%-4% of cases), gene amplification (2%-5%) and protein overexpression (2%-30%), with different prognostic and predictive outcomes. So far, non-selective tyrosine kinase inhibitors (TKIs) have shown a minor benefit in HER2-mutant NSCLC patients with objective response rates (ORRs) ranging from 0% to 19%. Trastuzumab-based chemotherapy was not found to be superior to chemotherapy alone [median progression-free survival (PFS) 6.1 versus 7 months, respectively] and dual HER2 antibody blockade with trastuzumab and pertuzumab had limited efficacy (ORR 13%-21%). In contrast, novel more selective HER2 TKIs such as poziotinib and pyrotinib have shown a promising activity in HER2-mutant pre-treated NSCLC patients, with response rates up to 38% and 44%, respectively. The most encouraging data come from phase II studies that evaluated the antibody-drug conjugates (ADCs) ado-trastuzumab-emtansine and trastuzumab-deruxtecan in patients with HER2-mutant NSCLC, with response rates of 50% and 62%, respectively. These agents are bringing hope to the management of HER2-altered NSCLC. Moreover, a paradigm shift from monotherapies towards combinations of agents with distinct mechanisms of action, such as ADCs with irreversible TKIs or immune checkpoint inhibitors, is already taking place and will change the therapeutic landscape of HER2-driven NSCLC. This paper provides a practical, concise and updated review on the therapeutic strategies in NSCLC with HER2 molecular alterations.

The McKesson prop--an essential tool for the emergency physician?

Facial trauma is a commonly encountered presentation to emergency departments. When associated airway compromise occurs, co-existing head and neck injuries serve to produce a challenging clinical situation. We describe two patients who suffered multi-system trauma, with severe maxillofacial injuries that necessitated prompt definitive airway management and mid-face stabilisation in the pre-hospital and emergency department phases of resuscitation. The McKesson prop is a simple yet highly effective tool for use in these injuries.

SEOM clinical guidelines for the treatment of small-cell lung cancer (SCLC) (2019).

Small-cell lung cancer (SCLC) accounts for 15% of lung cancers. Only one-third of patients are diagnosed at limited stage. The median survival remains to be around 15-20 months without significative changes in the strategies of treatment for many years. In stage I and IIA, the standard treatment is the surgery followed by adjuvant therapy with platinum-etoposide. In stage IIB-IIIC, the recommended treatment is early concurrent chemotherapy with platinum-etoposide plus thoracic radiotherapy followed by prophylactic cranial irradiation in patients without progression. However, in the extensive stage, significant advances have been observed adding immunotherapy to platinum-etoposide chemotherapy to obtain a significant increase in overall survival, constituting the new recommended standard of care. In the second-line treatment, topotecan remains as the standard treatment. Reinduction with platinum-etoposide is the recommended regimen in patients with sensitive relapse (≥ 3 months) and new drugs such as lurbinectedin and immunotherapy are new treatment options. New biomarkers and new clinical trials designed according to the new classification of SCLC subtypes defined by distinct gene expression profiles are necessary.

Contamination of salvaged maternal blood by amniotic fluid and fetal red cells during elective Caesarean section.

BACKGROUND: Cell salvage in obstetrics is still a controversial subject and has yet to be fully embraced. The aim of this exploratory study was to measure amniotic fluid (AF), heparin, and fetal red cell contamination of washed filtered salvaged maternal blood and to investigate differences based on the number of suction devices used. METHODS: Patients undergoing elective Caesarean section were assigned alternately to one of two groups. In Group 1, all blood and AF was collected with one suction. In Group 2, AF was aspirated to waste with a second separate suction device before collection of any blood. RESULTS: In both groups, alpha-fetoprotein (AFP), squames cells, and heparin were significantly reduced (P<0.001) by the washing and filtering process. Mean AFP levels post-filtration were 2.58 IU ml(-1) in Group 1 and 3.53 IU ml(-1) in Group 2. Squames cells were completely removed in all but two cases. Fetal red blood cells were still present in the final product, range 0.13-4.35%. In Group 1, haemoglobin and haematocrit were higher than in Group 2, with lower white blood cell, AFP, and fetal red cell counts. CONCLUSIONS: This study adds to the growing body of evidence that there is little or no possibility for AF contamination to enter the re-infusion system when used in conjunction with a leucodepletion filter.

T-regulatory cells are relatively deficient in squamous carcinomas undergoing regression in mice immunized with a squamous carcinoma vaccine enriched for immunotherapeutic cells.

In a prior report (Int J Cancer 2006; 119: 339-348), we described a new vaccination strategy for squamous cell carcinoma (SCC). The vaccine was prepared by transfer of unfractionated DNA-fragments (25 kb) from KLN205 cells, a squamous carcinoma cell line (DBA/2 origin; H-2(d)) into LM cells, a highly immunogenic mouse fibroblast cell line (C3H/He origin; (H-2(k))). As only a small proportion of the transfected cell population was expected to have incorporated DNA segments that included genes specifying antigens associated with the squamous carcinoma cells, we devised a novel strategy to enrich the vaccine for immunotherapeutic cells. Enhanced immunity to squamous carcinoma was induced in tumor-bearing mice treated solely by immunization with the enriched vaccine, which translated into prolonged survival without toxicity. Here, we describe the characteristics of the cell populations infiltrating established squamous carcinomas undergoing regression in mice immunized with vaccines enriched for immunotherapeutic cells. The results indicated that CD8+ T cells were predominant and that T-regulatory cells (FoxP3+, CD4/CD25+, CD4/CD62L(high), CD4/CTLA-4e) were relatively deficient in the regressing tumors. Inflammatory infiltrates were not detected in various organs and tissues of mice immunized with the DNA-based vaccine.

New strategy for the identification of squamous carcinoma antigens that induce therapeutic immune responses in tumor-bearing mice.

This study describes a new strategy for the identification of squamous carcinoma antigens tumor-associated antigens (TAA). The antigens were discovered by comparing microarrays of squamous carcinoma vaccines highly enriched for immunotherapeutic cells with non-enriched vaccines. The vaccines were prepared by transferring sheared genomic DNA fragments (25 kb) from KLN205 cells, a squamous carcinoma cell line (DBA/2 mouse origin (H-2(d)) into LM fibroblasts (C3H/He origin, H-2(k)). The transferred tumor DNA segments integrate spontaneously into the genome of the recipient cells, replicate as the cells divide and are expressed. As only a small proportion of the transfected cell population was expected to have incorporated DNA segments that included genes specifying TAA (the vast majority specify normal cellular constituents), a novel strategy was employed to enrich the vaccine for TAA-positive cells. Microarrays were used to compare genes expressed by enriched and non-enriched vaccines. Seventy-five genes were overexpressed in cells from the enriched vaccine. One, the gene for Cytochrome P450 (family 2, subfamily e, polypeptide 1) (Cyp2e1), was overexpressed in the enriched but not the non-enriched vaccine. A vaccine for squamous carcinoma was prepared by transfer of a 357 bp fragment of the gene for Cyp2e1 into the fibroblast cell line. Robust immunity, sufficient to result in indefinite survival, was induced in tumor-bearing mice immunized with cells transfected with this gene fragment.