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Background Cancer patients are at increased risk of death from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Cancer and its treatment affect many haematological and biochemical parameters, therefore we analysed these prior to and during coronavirus disease 2019 (COVID-19) and correlated them with outcome. Patients and methods Consecutive patients with cancer testing positive for SARS-CoV-2 in centres throughout the United Kingdom were identified and entered into a database following local governance approval. Clinical and longitudinal laboratory data were extracted from patient records. Data were analysed using Mann–Whitney U test, Fisher's exact test, Wilcoxon signed rank test, logistic regression, or linear regression for outcomes. Hierarchical clustering of heatmaps was performed using Ward's method. Results In total, 302 patients were included in three cohorts: Manchester (n = 67), Liverpool (n = 62), and UK (n = 173). In the entire cohort (N = 302), median age was 69 (range 19-93 years), including 163 males and 139 females; of these, 216 were diagnosed with a solid tumour and 86 with a haematological cancer. Preinfection lymphopaenia, neutropaenia and lactate dehydrogenase (LDH) were not associated with oxygen requirement (O2) or death. Lymphocyte count (P < 0.001), platelet count (P = 0.03), LDH (P < 0.0001) and albumin (P < 0.0001) significantly changed from preinfection to during infection. High rather than low neutrophils at day 0 (P = 0.007), higher maximal neutrophils during COVID-19 (P = 0.026) and higher neutrophil-to-lymphocyte ratio (NLR; P = 0.01) were associated with death. In multivariable analysis, age (P = 0.002), haematological cancer (P = 0.034), C-reactive protein (P = 0.004), NLR (P = 0.036) and albumin (P = 0.02) at day 0 were significant predictors of death. In the Manchester/Liverpool cohort 30 patients have restarted therapy following COVID-19, with no additional complications requiring readmission. Conclusion Preinfection biochemical/haematological parameters were not associated with worse outcome in cancer patients. Restarting treatment following COVID-19 was not associated with additional complications. Neutropaenia due to cancer/treatment is not associated with COVID-19 mortality. Cancer therapy, particularly in patients with solid tumours, need not be delayed or omitted due to concerns that treatment itself increases COVID-19 severity., Highlights • Pre-infection haematological/biochemical characteristics are not associated with COVID-19 severity. • Significant haematological/biochemical changes occur upon infection with heterogeneity in response observed. • High not low neutrophils were associated with oxygen requirement and COVID-19 mortality – GCSF should be used with caution. Age, haematological cancer, high neutrophil:lymphocyte ratio, CRP and low albumin were significant predictors of death in multivariable analysis. No significant complications requiring readmission were seen upon restart of cancer therapy following diagnosis of COVID-19.

Additional file 1 Figure S1 (A) Normalised signal of a logarithmic dilution of fixed iRFP expressing A431 cells suspended in low-melt agarose measured at two different times on the LPT: Before all other machines (first reading), and after all other machines (final reading). (B) Tumour signal or background signal per pixel per mouse on each day, measured on the LPT. Figure S2 (A-C) Tumour intensity images generated using our Mathematica MouseTensity software. Subcutaneous tumours from all 6 mice were imaged using all 3 platforms (A)XVI, (B)BIX, (C) LPT on day 17. The units for the x- and y-axes are pixels, and standardised units for the z-axis. Figure S3 (A-C) Tumour intensity images generated using our Mathematica MouseTensity software. Subcutaneous tumours from all 6 mice were imaged using all 3 platforms (A) XVI, (B) BIX, (C) LPT on day 24. The units for the x- and y-axes are pixels, and standardised units for the z-axis. Figure S4 (A) Example of saturated pixels detected in the XVI (Mouse 1 and 4). Only the saturated pixels are shown on the right and the tumour is shown on the left. Mouse 4, day 21 is representative of a low level of saturation, but this would still affect final quantification, while mouse 1 day 24 is representative of an obvious overexposure (B-D). Line graphs of the growth of subcutaneous tumours to track tumour growth in each mouse. These are the same data as Fig. 1E-G, but are now shown as lines, coloured per mouse. (E) Calliper measurements coloured to show growth of tumours over time. These are the same data as Fig. 1B, but are now shown as lines, coloured per mouse. Figure S5. (A) Examples of full body images of all mice on the last day of imaging on each platform. (B) Brightfield images of excised organs, taken in the LPT. Liver and lungs are shown for all mice, and front paw and knee that clearly showed a signal is shown for mouse 8. Figure S6. Fluorescent images of the knee and front paw of mouse 8 in all imaging platforms.

Additional file 2. Table S1 Details of the features of each tested platform, obtained from public data. Some information has been independently calculated/determined by the authors. Table S2 Image acquisition parameters for each platform for the plate, phantom and mice. Information in quotations is naming provided by software. Table S3 Statistics from Figure 1 EFG. Full statistical output from ANOVA, with Tukey comparisons. Table S4 Counts of tumours arising from tail vein injections, including percentages of the total number of tumours detected by each platform. The total tumour counts are listed in a separate table.

The application of precision medicine requires in-depth characterisation of a patient's tumours and the dynamics of their responses to treatment. We used next-generation sequencing of cfDNA to monitor therapy responses of a metastatic vaginal mucosal melanoma and show that cfDNA can be used to monitor tumour evolution and subclonal responses to therapy even when biopsies are not available., Background The application of precision medicine in oncology requires in-depth characterisation of a patient's tumours and the dynamics of their responses to treatment. Patients and methods We used next-generation sequencing of circulating cell-free DNA (cfDNA) to monitor the response of a KIT p.L576P-mutant metastatic vaginal mucosal melanoma to sequential targeted, immuno- and chemotherapy. Results Despite a KIT mutation, the response to imatinib was mixed. Unfortunately, tumours were not accessible for molecular analysis. To study the mechanism underlying the mixed clinical response, we carried out whole-exome sequencing and targeted longitudinal analysis of cfDNA. This revealed two tumour subclones; one with a KIT mutation that responded to imatinib and a second KIT-wild-type subclone that did not respond to imatinib. Notably, the subclones also responded differently to immunotherapy. However, both subclones responded to carboplatin/paclitaxel, and although the KIT-wild-type subclone progressed after chemotherapy, it responded to subsequent re-administration of paclitaxel. Conclusion We show that cfDNA can reveal tumour evolution and subclonal responses to therapy even when biopsies are not available.

Flow cytometry allows rapid multiparameter analyses of individual cells. Cells are illuminated with incident laser light of a specific wavelength. Resultant light scatter signals and fluorescence-emission signals from fluorochromes contained at the surface of the cell or within it are detected by an array of photomultiplier tubes. For example, fluorochrome-conjugated antibodies and DNA interchelating dyes can be used to acquire information on protein expression and DNA ploidy (via fluorescence signals) together with information on cell size and structure (via light-scatter signals).

Checkpoint kinase 2 (CHK2) is activated by DNA damage and can contribute to p53 stabilization, modulating growth arrest and/or apoptosis. We investigated the contribution of CHK2 to oxaliplatin-mediated toxicity in a colorectal cancer model.We evaluated the ability of CHK2 small molecule inhibitors to potentiate oxaliplatin-induced toxicity. The role of CHK2 in oxaliplatin-induced apoptosis was investigated in HCT116 cells that were wild-type (WT) or KO for CHK2. Small molecule inhibitors of CHK2 were used in combination studies with oxaliplatin in this cell model.In oxaliplatin-treated CHK2 KO cells, accelerated apoptosis was accompanied by attenuated p53 stabilization and p21(WAF-1) up-regulation correlating with increased Bax expression, cytochrome c release and elevated caspase activity. The higher levels of apoptosis in CHK2 KO cells were restored to control (WT) levels when CHK2 was re-introduced. This 'uncoupling' of p53 stabilization and Bax up-regulation in CHK2 KO cells suggested oxaliplatin-induced apoptosis was due to a p53-independent response. Combination studies revealed that CHK2 inhibitor II or debromohymenialdisine antagonized the responses to oxaliplatin. This inhibitory effect correlated with decreases in apoptosis, p53 stabilization and DNA inter-strand cross-link formation, and was dependent on the presence (but not activity) of CHK2.Combinations of CHK2 inhibitors with oxaliplatin should further sensitize cells to oxaliplatin treatment. However, these inhibitors produced an antagonistic effect on the response to oxaliplatin, which was reversed on the re-introduction of CHK2. These observations may have implications for the use of oxaliplatin in colorectal cancer therapy in combination with therapies targeting CHK2.

Progress in the prevention and treatment of women at risk of pre-eclampsia (PE) still remains hindered by the lack of clinical screening tools that can accurately predict which mothers are at risk. The identification and validation of predictive biomarkers is therefore seen as a critical milestone towards improved healthcare provision and the clinical testing of new therapeutic strategies. Gel-free proteomic technologies offer the capability of analysing hundreds of plasma proteins simultaneously, but as yet these methods have not been applied to pregnancy complications. To assess the feasibility of such an approach to plasma biomarker research in pregnancy we have applied the technique to samples from women with PE to gestation-matched controls.Pooled plasma samples taken at time of disease from women with PE (n = 23) and gestation-matched controls (n = 23).Proteomics strategy for relative quantification of proteins using mass spectrometry.We identified several differences, including elevated levels of endoglin, PAPP-A and PSG1 in PE plasma. Increased levels of endoglin were validated using immunoassay analysis of individual plasma samples.Although at a relatively early stage, this mass spectrometry-based approach shows promise as a tool to identify global protein changes in plasma. The application of these methods to pre-disease samples is the next step in the identification of clinically useful biomarkers.

We describe the results of using a free cartilage graft in the closure of cleft palate fistulae in 14 patients with a mean follow-up of 8.6 months. Complete closure of the fistula was achieved in 11 patients (79%), with partial closure in the remaining three patients. This technique is simple, causes relatively little discomfort, involves little tissue dissection and can be performed as a day-case procedure. The success rate is comparable with or better than other methods, and we consider it the treatment of choice for small cleft palate fistulae.

We report a new case of S.C.C. of the large bowel with multiple liver metastases. A resection of the primary tumour and liver biopsies were performed with administration of a postoperative chemotherapy (5-Fluorouracil). After a stabilization of 3 months, the metastases were rapidly progressive and the patient died a year after the diagnosis. About 70 cases of S.C.C. of the colon and rectum have been described in the literature. It is most common in the fifth decade and occurs equally in male and female. The most frequent locations are the rectum and the sigmoid. Clinical and physical features and common diagnostic methods do not differentiate the S.C.C. from adenocarcinoma. Treatment is the same but the prognosis of S.C.C. appears to be worse than that of adenocarcinoma.

We describe an improved technique which allows the analysis of enzyme reaction kinetics for gamma-glutamyl transpeptidase (gamma-GT) by flow cytometry. This is technically difficult because of the location of the enzyme on the external surface of the cell membrane leading to the rapid escape of the product. The reaction is determined by monitoring the conversion of gamma-glutamyl aminomethylcoumarin to aminomethylcoumarin. Reaction kinetics are described for BL8 hepatocyte and JB1 hepatoma cells lines, together with inhibition kinetics for the active site-directed glutamine analogue L-(alpha-S,5S)-alpha-amino-3-chloro-4,5-dihydro-5-isoxazoleacetic acid. We show that it is possible to follow the reaction dynamics in a heterogeneous mixture of BL8 and JB1 cells allowing discrimination of the two cell types based on gamma-GT activity. Improvements for further optimizing the assay of this important enzyme are suggested.

Many anticancer agents induce an active cell death process, apoptosis, in sensitive tumour cells. Elucidation of molecular mechanisms underlying apoptosis may shed light on why some tumour cells survive chemotherapy, and may identify new targets for anticancer agents whose effects are not tightly linked to proliferative status. The signal transduction events which initiate apoptosis are unclear. A change in cytosolic calcium is generally assumed to be a key signal for apoptosis although the evidence for this is not conclusive. Other putative signal transducers which may modulate apoptosis are protein kinase C and cAMP. Genes which induce apoptosis in response to such signals are largely unidentified, but certain oncogenes, notably bcl-2, act to delay or suppress apoptosis in several cell types.

Functional and structural changes accompany the differentiation of granulosa cells during follicular development. We used flow cytometry and fluorescent dyes to characterize two organelles important to the steroidogenic process. Mitochondria, which contain the rate-limiting enzyme responsible for cholesterol conversion to pregnenolone, and lipid droplets, which store cholesterol substrate, were probed in viable hen granulosa cells during differentiation. The fluorescent dye Dio3-C5 (DiO) was used to probe mitochondrial membrane potential, indicative of mitochondrial activity and/or number, during rapid granulosa cell differentiation in a hierarchy of individual developing hen preovulatory follicles (F6, smallest, to F1, largest). Cellular DiO fluorescence, granularity, and cell size were significantly elevated with increasing maturation state. Treatment with LH significantly increased DiO fluorescence in granulosa cells from F1 but not F3. The increased mitochondrial activity/number in granulosa cells that accompanies follicular maturation and is influenced by LH may reflect, at least in part, increased activity or amount of hormone-regulated mitochondrial enzymes controlling steroidogenesis. Flow spectrofluorometry and the metachromatic lipid dye, nile red, were used to probe lipid droplets in differentiating granulosa cells from F6 to F1. There was a dramatic increase in the fluorescence component related to lipid droplets with increasing stages of follicular maturation, suggesting recruitment of lipids into droplets during the differentiation of granulosa cells into hormone-responsive steroidogenic cells. The results demonstrate the dynamic nature of the granulosa cell morphology involved in steroidogenesis during follicular development.(ABSTRACT TRUNCATED AT 250 WORDS)

Defecatory difficulties may be a source of great distress to patients. Normal defecation requires the coordination of abdominal and pelvic muscles. Throughout the last decade, many studies have demonstrated various and often mixed abnormalities of these mechanisms. Pathophysiological studies are able to determine, in a specific patient, the most probable causes of the obstructive symptom. Progress in the management of such disorders can only be achieved by understanding their pathophysiology.

The authors review the recent literature about the classification of primary motor disorders of the oesophagus: achalasia, diffuse oesophageal spasm, nutcracker oesophagus, hypertensive lower oesophageal sphincter and non-specific intermediary disorders. In fact these motility disorders belong to a spectrum of diseases closely related, with chronological transformation of a specific disorder to another one or to intermediary disorders. Most of the recent pathophysiological research concerned primary achalasia, secondary achalasia syndromes being a kind of experimental model. These studies point to a morphological or functional deficiency of postganglionic nerves inhibiting the lower oesophageal sphincter (LOS) through noncholinergic nonadrenergic neurotransmitters. Recent advances in the treatment of achalasia and other motility disorders are not yet based on these findings. Although calcium channel blockers, like nifedipine, lower LOS basal pressure, they are not very useful on long term relief of symptoms of achalasia. Pneumatic dilatations or cardiomyotomy still remain the best methods of treatment of this disease.

The authors review the recent literature about the methodology of oesophageal pH monitoring, which has a high sensitivity and a high specificity for assessing gastro-oesophageal reflux. Combined electrodes offer most advantages. Ambulatory recording should be done in hospital under standard conditions (meals), particularly for clinical studies. The best clinical indication is to detect pathological reflux in case of atypical symptoms with negative oesophagoscopy. The authors give their normal values in a series of measurements with two systems as well as their results in a series of cases of oesophagitis of various grades.

Patients with gastroparesia or intestinal pseudo-obstruction frequently have a motility disorder of the antro-pyloric region that makes the passage of tubes through the pylorus impossible or a very long procedure. We describe a rapid endoscopic method, inspired by the one used for nasobiliary drainage, that simplifies this procedure. In our manometric studies, we observed that spontaneous positioning needed a mean of 57 min (SD 34) in nondyspeptic subjects and a mean of 147 min (SD 68) in non-organic highly dyspeptic patients. Our method allowed us to shorten to a mean of 22 min (SD 5) the time needed to position the manometric tube and to start the recording of gastro-intestinal contractions in highly dyspeptic patients.

SUMMARY We studied the local humoral immunity of histologically non-involved jejunum in patients with Crohn's disease. Normal subjects and patients with ulcerative colitis served as controls. Jejunal fluid samples were collected during segmental jejunal perfusion, under an occluding balloon and the in vivo jejunal secretion of the following proteins was determined: albumin, orosomucoid, transferrin, α2-macroglobulin, secretory component, monomeric and polymeric IgA, IgG, and IgM. The densities and number of IgA-, IgG-, and IgM-containing cells in the lamina propria of the jejunum were measured on perendoscopic biopsies. Jejunal secretion of polymeric IgA and the density of IgA-containing plasma cells in the lamina propria were significantly lower in patients with Crohn's disease than in both control groups. This abnormal intestinal immune response, which was not correlated to the activity of the disease, might be involved in its pathogenesis.

We studied digestive manifestations occurring during follow-up of 19 cases of myotonic dystrophy. GI symptoms occurred in 10 patients (53%), mainly dysphagia (n = 7). In patients undergoing investigations (n = 7), their digestive troubles were probably or definitively related to the neurological disease in 6 cases. Upper GI endoscopy was normal in 5 dysphagic patients. The oesophageal manometry was abnormal in 3 of 4 patients. Complications developed in 3 cases (16%). One patient presented two episodes of spontaneous pneumoperitoneum. A second one developed a chronic colonic pseudo-obstruction with megacolon treated by subtotal colectomy. Manometric studies revealed oesophageal aperistalsis, low amplitude of gastro-jejunal contractions but persistence of migrating motor complexes and a normal recto-anal reflex. A third patient developed an acute pseudo-obstruction with ischemic colitis. In conclusion, myotonic dystrophy frequently disturbs digestive tract motility and may cause complications such as intestinal pseudo-obstruction.

Elevated antibody (Ab) levels to gliadin (GL), the alcohol soluble fraction of gluten, are present in the serum of patients with coeliac disease (CD), an enteropathy caused by hypersensitivity to gluten. The site of production and role of these Abs in the pathogenesis of the disease are poorly understood. In order to evaluate in vivo the secretion of Ig by the jejunal mucosa in CD, we have analysed jejunal fluids collected during a perfusion of a 40cm jejunal segment under an occluding balloon which avoid contamination with other intestinal secretions (1). Diisopropylfluorophosphate (protease inhibitor) was added to the samples Jejunal biopsies and serum were also collected.

Background: The X-linked inhibitor of apoptosis (XIAP) is a potent anti-apoptotic protein. AEG35156 is a 2nd generation antisense oligonucleotide to human XIAP that enhances cancer cell apoptosis preclinically as a single agent and in combination with chemotherapeutics. Methods: The primary objective was to establish the maximum tolerated dose of AEG35156 given as a 2-hour infusion once weekly following 3 daily loading doses in week 1. Other objectives were to evaluate AEG35156 pharmacokinetics, XIAP inhibition, circulating apoptosis biomarkers and document anti-tumour activity. Results: Twenty-nine adult patients (16 M; 13 F) have been registered. Twenty-three have completed at least one 3-week treatment cycle at doses ranging from 60 to 500mg AEG35156. Treatment has been generally well tolerated, the most common toxicities have been grade I chills/pyrexia, rapidly reversible increases in aPTT at the end of infusion and mild transaminitis. One patient at 240mg had a silent myocardial infarction probably related to prophylactic aspirin withdrawal prior to starting protocol therapy. Three DLTs were observed at AEG35156 350mg: Grade 3 chills/lethargy (mesothelioma), grade 3 transaminitis (colon), transient grade 3 chills/tumour lysis syndrome/transaminitis (small lymphocytic lymphoma). A less intensive dosing protocol was deemed appropriate for future NHL patients and a protocol amendment excluding NHL patients from this study and permitting resumption in patients with solid tumors and including premedication with dexamethasone / acetaminophen was implemented. Dosing at AEG35156 500mg has been completed with no DLTs and the study is ongoing at 650mg. Pharmacokinetics were dose-proportional and linear from 60 to 500mg AEG35156. Mean plasma Cmax at 500mg was 55�g/mL. Decreases in XIAP mRNA have been observed in PBMCs at dose levels >240mg and increased levels of circulating biomarkers of apoptosis have been observed in some patients. One patient with NHL who received only a single infusion had rapid clearance of circulating tumour cells and reduction in nodal disease. Conclusions: AEG35156 can be safely delivered by 2-hour infusions and preliminary evidence of XIAP mRNA knockdown and antitumour activity in lymphoma has been observed.

The sera of 15 patients with liver disease and high serum concentrations of secretory component (SC) were analyzed by density gradient ultracentrifugation and radioimmunoassays for SC, IgA, and IgM to determine the molecular state of SC and IgA. Results in serum were compared to those obtained by simultaneous analysis of bile in five of the patients, and to those in serum and bile of a case of total IgA deficiency. Free SC was virtually not found in sera, although it was well represented in all bile, up to 88 and 97% of total bile SC in a case of complete biliary obstruction and in IgA deficiency, respectively. IgM-bound SC was found in all sera, amounting to 91% and 100% of total serum SC, respectively, in a case of acute hepatitis with a very high serum IgM concentration and in IgA deficiency. The proportions of SC bound to IgM and to polymeric IgA (p-IgA) in the sera correlated with their IgM/p-IgA molar ratio. This suggests that during liver disease, the hepatobiliary tissues could release free SC into the circulation, where it binds to p-IgA and IgM according to their respective concentrations and affinities for SC. The proportion of p-IgA in serum was not increased in four cases of biliary obstruction, in contrast to our cases of cirrhosis and acute hepatitis, indirectly supporting a minor transfer of p-IgA from blood to bile in humans, in contrast to rats and rabbits.

The influence of insulin on the postnatal development of intestinal functions linked to villus cells (sucrase, lactase, maltase and aminopeptidase) and crypt cells (secretory component of immunoglobulins, SC) has been studied in suckling and weanling rats. At 9 days of age, the animals received a daily injection of insulin 12.5 mU g-1 body weight day-1 for 4 days. Compared with saline-treated controls, insulin had no effect on the development of the intestinal mucosal mass parameters determined in the jejunum, ileum and colon. A premature appearance of sucrase was noted in isolated jejunal villus and crypt cells, the level of activity reached by the enzyme being dependent of the amount of insulin injected. By 6 and 12 h after a single injection of the hormone (12.5 mU g-1 body weight), sucrase activity was detected in all the cell fractions along the villus-crypt axis. In villus cells of insulin-treated rats, maltase, lactase and aminopeptidase activities were significantly (P less than 0.001) increased (+201%, +50%, +207%, respectively, vs. controls), whereas the concentration of SC measured by a sensitive immunoradiometric assay was enhanced over the controls by 75% in villus cells, 83% in crypt cells and 172% in the liver. Weanling rats treated from day 10 to day 20 postpartum with 12.5 mU insulin also exhibited a higher intestinal production of SC (+93%, P less than 0.01) than did saline controls.(ABSTRACT TRUNCATED AT 250 WORDS)

The incidence, titer and molecular size of anti-gliadin (GL) IgA were analyzed in sera from children with celiac disease (CD, n = 66), hospitalized control children (n = 64) and children with other malabsorption disorders (n = 103). Anti-GL IgA was assessed by solid-phase radioimmunoassay, and its molecular size was analyzed by sucrose density gradient ultracentrifugation. The median anti-GL IgA titer was significantly higher in untreated CD than in the other groups, with some overlap. Whereas 57% of anti-GL IgA were polymeric IgA (pIgA) in acute CD, only 17% were pIgA in children with other malabsorption disorders. These anti-GL pIgA consisted mainly of dimers with in some children, a low proportion of anti-GL secretory IgA (SIgA). A large proportion of anti-GL pIgA was associated with high total serum IgA. After the onset of a gluten-free diet (GFD), anti-GL pIgA disappeared more quickly from serum than mlgA. During the gluten challenge test, anti-GL IgA comprised 20% pIgA when they reappeared in serum, but 59% pIgA at the end of the test. The presence of anti-GL pIgA in serum seems related to chronic exposure to gluten of CD patients with a flat intestinal mucosa. These serum pIgA antibodies could result from a spillover from the intestinal mucosal synthesis into the circulation.