A significant difference was also observed between the TAO groups (P < 0·05). Figure 2 shows the values of the determinations of Th1 cytokine profiles (IFN-γ

and IL-12) in the plasma of normal individuals (smoker, ex-smoker and non-smokers) and patients with TAO (smokers and former smokers). The data results show an increase of these cytokines in the plasma of TAO patients compared with control subjects (P < 0·05 for each comparison). A significant buy Kinase Inhibitor Library difference was also observed between the TAO groups (P < 0·05). Figure 3 shows values of the determinations of Th2 cytokine profiles (IL-4, IL-10, IL-13 and IL-5) in the plasma of normal individuals (smoker, ex-smoker and non-smokers) and patients with TAO (smokers and former smokers). The data results show increased levels of IL-4, IL-5 and IL-13 in the plasma of TAO patients compared with control subjects (P < 0·05 for each comparison). Decreased levels of IL-10 were found in patients with TAO active smokers compared to control individuals and TAO former smokers (P < 0·05 for each comparison). Figure 4 shows the values of the determinations of Th17 cytokine profiles (IL-17 Raf inhibitor and IL-23) in the plasma of normal individuals (smoker, ex-smoker and non-smokers) and patients with TAO (smokers and former smokers). The data results show an increase of these cytokines in the

plasma of TAO patients compared with control subjects (P < 0·05 for each comparison). Because the development and aetiology of TAO have not yet been elucidated, and as the direct action of inflammatory mediators has been observed in the vascular endothelium of TAO patients, in this study we have evaluated some components of the cytokines in the

plasma of TAO patients who presented with acute symptoms. To the best of our knowledge, this is the first complete investigation including cytokines with proinflammatory, Th1, Th2 and Th17 profiles. The precise cause of TAO Tryptophan synthase is still unknown, and different hypotheses have been suggested. A reaction to the constituents of cigarettes is recognized as a factor in the initiation, progression and prognosis of this disease. It is possible that genetic modifications or autoimmune disorders are implicated [5,12,13]. Thus, the strong relationship with smoking seems to involve direct toxicity to the endothelium by certain tobacco products (nicotine) or an idiosyncratic immune response to some agents. Most patients with TAO have hypersensitivity to extracts of tobacco. Peripheral endothelium-dependent vasodilation is impaired in the non-diseased limbs of TAO patients, and this vascular dysfunction may contribute to such characteristics as segmental proliferative lesions or thrombus formation in the peripheral vessels [14]. The immune system seems to play a critical role in the aetiology of TAO.

5%) to the second one (12.8%). Conclusion: Routine use of IV heparin following digital replantation and revascularization is not warranted. Surgical technique and

type of injury remains the most important predictors for success in selleck these complex procedures. © 2011 Wiley-Liss, Inc. Microsurgery 2011. “
“The patient was a 62-year-old man with chief complaints of pharyngeal pain and dysphagia. He was diagnosed with pyriform sinus poorly differentiated squamous cell carcinoma T3N0M0 (Stage II) and underwent partial laryngopharyngectomy, lymphadenectomy in the right neck, tracheostomy, and reconstruction of the larynx and aryepiglottic fold with a free radial forearm flap and the associated vascularized palmaris longus tendon. No particular problems occurred after surgery, and swallowing and articulation functions were successfully recovered. A free jejunum transfer is the first choice for reconstruction

of a defect after partial hypopharyngectomy, but reconstruction of the supracricoid complex structure of the larynx using a free jejunum transfer after partial laryngopharyngectomy may lead to aspiration of intestinal fluids. In this case, we performed functional reconstruction of the laryngopharyngeal defect using a free radial forearm flap including a vascularized tendon of the palmaris longus, and satisfactory postoperative function was achieved. We believe that the key to successful functional recovery after partial laryngopharyngectomy is establishment of the three-dimensional complex structure of the arytenoid and aryepiglottic fold. © 2011 Wiley Periodicals, Inc. Microsurgery, 2012. “
“The purpose of this study is to evaluate Selleck Mitomycin C the use of the venous anastomotic Flow Coupler in monitoring free flaps used for breast reconstruction in a consecutive series of patients. Retrospective data were 5-Fluoracil purchase collected on patients undergoing free flap breast reconstruction from May 2012 to March 2014. The venous anastomotic Flow Coupler was used in the first 85 flaps and a non-flow

Coupler with clinical and external Doppler monitoring alone in the subsequent 34 flaps. Data collected included patient age, BMI, prior radiation, flap type, intra- and postoperative Flow Coupler events, along with rates of flap take back, salvage, and failure. Proportion data were compiled and statistically analyzed. One hundred nineteen consecutive abdominal based breast reconstruction free flaps were performed. The overall flap failure rate was 4.2% (4.7% Flow Coupler; 2.9% in non-flow Coupler; P = 1.0). The Flow Coupler demonstrated 100% sensitivity in the intra- and postoperative settings. A positive predictive value of 36% was noted intraoperatively which was significantly higher compared to the non-flow Coupler group (P = 0.015). Vessel thrombosis occurred in 17.6% of Flow Coupler flaps, which was significantly higher when compared to the non-flow Coupler (2.9%; P = 0.038). The Flow Coupler is a sensitive method to confirm patency of a microsurgical anastomosis.

[4] It has been demonstrated that allergens in the presence of

endotoxins trigger a substantially stronger allergic inflammation, compared with that evoked in the absence of endotoxins.[5-7] After inhalation, endotoxins, such as lipopolysaccharide (LPS), encounter and activate alveolar macrophages, leading to the production and release of pro-inflammatory cytokines, chemokines, adhesion molecules and other mediators.[8] Nasal and lung lavage samples of allergic subjects show increased levels of interleukin-1β (IL-1β),[9] primarily produced by activated macrophages.[10] Production Selleckchem AZD8055 of mature IL-1β requires distinct signals, some of which induce gene expression in the so called ‘priming step’, whereas other signals trigger the maturation of pro-IL-1β to IL-1β by a multiprotein complex called inflammasome. The NLRP3 inflammasome complex consists of NLRP3 (NOD-like receptor family pyrin domain-containing 3) sensor, caspase-1 and ASC (apoptosis-associated speck-like protein containing a caspase recruitment domain) adaptor.[11, 12] NLRP3 inflammasomes play a crucial role in the detection and sensing of exogenous danger signals like pathogen-associated molecular patterns and toxins of microbes, asbestos or silica, as well as endogenous danger signals like monosodium urate and amyloid.[13, 14] Most NLRP3 activators have been shown to induce ROS Epigenetic Reader Domain inhibitor generation,[15]

and unless inhibitors of ROS production or ROS scavengers attenuate NLRP3 inflammasome activation[16] implying an essential role for ROS in NLRP3 function. As pollen NADPH oxidases are able to generate ROS, and ROS have been implicated in the NLRP3 inflammasome-mediated IL-1β production, we hypothesized that exposure to pollen extract may influence inflammatory responses and IL-1β production of macrophages via NLRP3 inflammasome. Here we report for the first time that ragweed

pollen extract (RWE), typically used as a model for pollen action,[3] significantly elevates LPS-induced IL-1β production of THP-1 or primary macrophages and dendritic cells in an NADPH-dependent manner. We also demonstrate that a caspase-1 inhibitor or NLRP3 silencing abolish this enhancing effect together with the original LPS-triggered inductions. We also show that RWE in the presence of NADPH enhances LPS-induced p38 and Jun N-terminal kinase (JNK) signalling pathways resulting in the activation of AP-1 transcription factors and the subsequent gene transcription/expression of pro-IL-1β and key components of the inflammasome. This effect is mediated by a ROS-dependent mechanism. The THP-1 cell line (ATCC TIB-202) was a generous gift from Professor Laszlo Nagy. THP-1 monocytes were cultured in RPMI-1640 (Gibco BRL Inc., Grand Island, NY) containing 10% heat-inactivated fetal calf serum, penicillin-streptomycin and glutamine, and maintained at 37° under 5% CO2.

Therefore, the loxP insertions

at 143 nt and 191 nt decreased the viral packaging efficiency. Adenovirus vectors can efficiently transduce a transgene not only in vitro, but also in vivo (1–4). First-generation AdV can be amplified only in 293 cells, a cell line containing Ad5 E1 DNA in its genome, because the E1 region, an essential region for the virus, is substituted for a transgene. However, first-generation AdV is problematic in that it induces immune responses against small amounts of expressed virus protein(s) of unknown origin (5–8). To solve this problem, the use of a helper-dependent (HD)-AdV has attracted attention (7, 9, 10). With HD-AdV, no virus proteins are expressed because all the viral coding regions LBH589 datasheet are substituted for foreign sequences; only the ITR, comprised of 102 nt at both ends of the virus genome, and the packaging domain, located RXDX-106 nmr within the left 0.4 kilobases in the Ad5 genome, are retained. To amplify the HD-AdV, a helper virus that retains most of the virus genome and supplies the viral gene products is used. To avoid contamination with the helper virus during HD-AdV preparation, the packaging domain of the helper virus is flanked by a pair of target sequences for a site-specific recombinase: loxP of Cre, derived from bacteriophage P1 (11,12),

or FRT of FLP, derived from the 2- μm plasmid of Saccaromyces cerevisiae (13–15). Because the site-specific recombinase mediates the excisional deletion of the DNA sequence flanked by the pair of

target sequences, the packaging of the helper virus is hampered in recombinase-expressing 293 cells by the specific excision of the packaging domain from the helper virus genome, enabling the packaging of the HD-AdV genome into a virus capsid to be prioritized. However, Dichloromethane dehalogenase the removal of the packaging domain is not perfect, and some helper viruses still containing the packaging domain always remain (7, 9, 16, 17). This observation prompted us to examine the influence of the loxP insertion on the packaging efficiency in E1-deleted AdV, including the helper virus of HD-AdV. The packaging domain of Ad5 has well been characterized (18–22). The cis-acting packaging domain is reportedly located between 194 nt and 380 nt from the left end of its genome and overlaps with the E1A enhancer region (18, 23). The domain contains seven repeated sequences (termed A-repeats), of which AI, AII, AV and AIV are the most important for packaging activity and contain a consensus motif, 5′-TTTGN8CG-3′ (19). Because the insertion sites of both the loxP are close to the packaging domains, these insertions may affect the virus titer of the helper virus. Previously, the sites of loxP-insertion downstream of the packaging domain were reported to influence the packaging of the helper virus (24) and the efficiency of the production of HD-AdV (25).

Despite conventional and empirical treatments, the patient developed progressive neurological deterioration leading to death. Autopsy showed Primary angiitis of the CNS (PACNS) with predominant cranial neuropathy, spinal cord involvement

and extensive myelomalacia. “
“Meningiomas are the most common primary intracranial tumors. They are usually benign and slowly growing; however, they may show histologically malignant features categorizing them into grade II or III of World Health Organization (WHO) classification. Rhabdoid meningioma (RM) is an uncommon meningioma variant categorized as WHO grade III. The clinical course of RM is determined by local recurrences, invasion of adjacent brain and/or dura, widespread leptomeningeal

dissemination, remote buy Idasanutlin metastases and fatal clinical outcome. Herein we report a case with recurrent aggressive left occipital parasagittal region RM in which the patient initially declined radiation treatment. The tumor was resected four times in 5 years. Histopathological examination revealed a rhabdoid meningioma with metaplastic, papillary and chordoid differentiation. Six months after her fourth operation the patient died of progressive disease. RM is a rare subtype of malignant meningioma and the role of different adjuvant therapeutic options are still unknown. Clinical presentation, radiological features and pathologic findings of this uncommon tumor are discussed. “
“K. Morgan (2011) Neuropathology and Applied Neurobiology37, 353–357 The three new pathways leading to Alzheimer’s disease Genome-wide association studies (GWAS) promise a significant impact on the understanding of late-onset Alzheimer’s LDK378 purchase disease (LOAD) as the genetic components have been estimated to account for 60–80% of the disease. The recent publication of results from large GWAS suggests that LOAD is now one of the best-understood complex disorders. Four recent large

LOAD GWAS have resulted in the identification of nine novel loci. These genes are CLU– clusterin, PICALM– phosphatidylinositol-binding clathrin assembly protein, CR1– complement receptor 1, BIN1– bridging integrator 1, ABCA7– ATP-binding cassette transporter, MS4A cluster – membrane-spanning 4-domains subfamily A, CD2AP– CD2-associated Staurosporine order protein, CD33– sialic acid-binding immunoglobulin-like lectin and EPHA1– ephrin receptor A1. Collectively, these genes now explain around 50% of LOAD genetics and map on to three new pathways linked to immune system function, cholesterol metabolism and synaptic cell membrane processes. These three new pathways are not strongly linked to the amyloid hypothesis that has driven so much recent thinking and open up avenues for intensive research with regard to the potential for therapeutic intervention. “
“Materials from our first autopsied case of diffuse Lewy body disease (DLBD), that was originally reported in 1976, were re-examined using recent immunohistochemical methods.

The fifth gene, located on scaffold_45 (Emoal for oncosphere-antigen-like; position 4212–3089) represents a novel, distantly related member of the EG95/45W family that has not yet been described in studies on vaccine development (Figure 4). Very much like EM95, Emoal is specifically expressed in regenerating primary cells; it displays an exon–intron structure that is typical for the EG95 gene family, and its gene product comprises a signal peptide, one Fn3 domain and a C-terminal transmembrane domain, suggesting that it has a similar function as the EG95/45W proteins

described so far. A close ortholog to X-396 Emoal, Egoal, is also present on the genome of E. granulosus (contig_32513; position INCB024360 in vivo 4699–3576), which could prove important for the further development and improvement of vaccine formulations against CE. Interestingly, and in contrast to the AgB family, the genome of H. microstoma is absolutely free of EG95/45W-like sequences, which supports the idea that this gene family is indeed highly specific to taeniid tapeworms. In addition to the TSOL18 and TSOL45 antigens of T. solium, extensive vaccination trials against porcine cysticercosis have already been undertaken using the so-called S3Pvac vaccine (114,115). S3Pvac consists of three synthetic peptides (named KETc12, KETc1, GK1) that had been identified by immune-screenings

against T. crassiceps cDNA libraries and when tested under field conditions, SP3vac could reduce the number of T. solium infected pigs by 50% and lowered parasite load by >90% (90). Interestingly, in spite of the fact that a considerable amount of information has already been published on S3Pvac (90), including a recent report on the presence of similar sequences in other cestodes (116), the proteins and genes which correspond to the synthetic peptides have never been characterized so far. We therefore analysed the situation for E. multilocularis using the published KETc1 and GK1 sequences as well as E. multilocularis PJ34 HCl genome and transcriptome data. The GK1 peptide clearly maps to the amino acid sequence

encoded by a predicted gene on scaffold_13 (position 1.570.711–1.568.292). The encoded protein (264 amino acids; 29 kDa; Figure 6) contains one Glucosyltransferase/Rab-like GTPase activators/Myotubularin domain (GRAM domain), which is thought to be an intracellular protein-binding or lipid-binding signalling domain, and one WWbp domain which is characterized by several short PY- and PT-motifs and which presumably mediates tyrosine phosphorylation in WW domain–ligand interactions (Figure 6). At least within the WWbp domain, this protein displays significant homologies (47% identical, 68% similar residues) to a predicted S. mansoni protein, WW domain-binding protein 2 (accession no. FN313948), of unknown function.

Furthermore, analysis of serum anti-HAF antibody isotypes, mesangial immune deposits and splenocyte interferon (IFN)-γ, monocyte chemoattractant protein-1 (MCP-1) and regulated upon activation normal T cell expressed and secreted (RANTES) secretions indicated that CpG-DNA induced a T helper type 1 (Th1) response in mice with HAF-GN. Previously, we reported that monovalent targeting of FcαRI strongly inhibited the development of immune complex-induced GN through decreased macrophage infiltration [16]. Therefore, we hypothesized that FcαRI

selleck products targeting should control the harmful immune complex HAF-CpG-GN model mediated by TLR-9 signalling. We found that monomeric occupancy of FcαRI alleviated the worsening glomerular damage triggered by TLR-9 activation. These results suggest that shifting the inflammatory balance by specifically targeting FcαRI could represent a new viable option for the

treatment of severe renal inflammatory diseases. The mice were bred and maintained in the mouse facilities of the Research Institute for Diseases of Old Age (Juntendo University School of Medicine, Tokyo, Japan). CHIR-99021 mw All experiments were conducted in accordance with national guidelines. A construct encoding human FcαRIR209L/FcRγ-FLAG was obtained by inserting a 1165-base pairs (bp) cDNA fragment into the Escherichia coli strain RI (EcoRI) site of a CAG promoter

containing β-actin (UniTeck, Kashiwa, Japan). Three progeniture lines Forskolin supplier were found to contain the human FcαRIR209L/FcRγ-FLAG cDNA by polymerase chain reaction (PCR) of tail DNA using transgene-specific primers 5 9-GGGTCATTAGTTCATAGCC-3 9 and 5 9-GGCATATGATACACTTGAT- 3 9. The C57BL/6J background was introduced into line 604 by more than eight consecutive crosses. All mouse strains in this study were bred and housed in strictly controlled specific pathogen-free conditions. We prepared the FcαRIR209L/FcRγ transfectant (I3D) from a mouse macrophage cell line (RAW264·7) using the Cell Line Optimization Nucleofector Kit (Lonza, Walkersville, MD, USA). The mouse macrophage cell line RAW264·7 was cultured in Glutamax (Invitrogen, Carlsbad, CA, USA) supplemented with 10% fetal calf serum (FCS), 100 U/ml penicillin and 100 µg/ml streptomycin at 37°C with 5% CO2 in a humidified incubator. Stable transfectants in the presence of Geneticin (1·0 mg/ml; Sigma-Aldrich Chemicals, Steinheim, Germany) were selected.

In contrast, our knowledge of the burden and impact of anxiety disorders, lower perceived social support and impaired HRQOL in these patients is limited. Further studies using standardized diagnostic criteria are required. The proposed mechanisms by which psychosocial factors influence the clinical course of see more CKD also require elucidating and may provide targets for clinical intervention. In Australia, current clinical practice guidelines advocate the provision of educational

information regarding the psychological aspects of CKD for both pre-dialysis and dialysis patients.[51] However, there are currently no existing recommendations to guide the routine assessment of psychosocial factors and HRQOL. Effective assessment and intervention will require considerable resources and integration of patient care involving physicians, nurses, social workers, mental health professionals and family members. Innovative client and family focused models of care in which patients are supported and encouraged to improve health literacy, capability and autonomy may be efficacious;[52] however, high level clinical evidence is required. Data from Canada indicate that the economic benefits of delaying the disease progression of CKD may more than compensate

for the additional costs of implementing a multidisciplinary model.[53] This review highlights the need for methodologically robust prospective studies to assess the burden and relative influence of psychosocial factors and HRQOL on outcomes at different

stages of CKD. This has the potential to provide BIBW2992 an evidence base for revising healthcare provision in order to optimize the clinical care and reduce the public health burden of this growing patient population. “
“Since the introduction of haemodialysis in the management of acute kidney injury in the 1940s and for chronic kidney disease (CKD) in the 1960s dialysis has become one of the most successful advances in medical technology, with almost 11 000 patients Tenofovir nmr currently receiving dialysis in Australia and almost 2500 in New Zealand. Like all medical technologies, its place continues to evolve. For a time, dialysis was seen as a treatment best delivered only to younger patients without diabetes; today the greatest uptake of dialysis is in patients over age 65 and the most common cause of needing dialysis is diabetes. Along with these extended criteria for dialysis, that have evolved over many years, has come the recognition that the older dialysis patient often has considerable co-morbidity and frailty, that time spent on dialysis is not always beneficial to these patients and that their overall prognosis is considerably worse than their younger counterparts. CARI guidelines recommend that ‘an expectation of survival with an acceptable quality of life’ is a useful starting point for recommending dialysis.

Results: S-40542 displayed twofold higher affinity to androgen receptor than bicalutamide in vitro. Subcutaneous repeated administration of S-40542 (10–100 mg/kg) significantly reduced the prostate weight. Oral repeated treatment with S-40542 (30, 100 mg/kg) for 28 days significantly suppressed selleck compound growth of KUCaP-2 tumor. Similar administration of bicalutamide also exerted significantly anti-tumor effect in the model. The serum prostate-specific antigen level was little influenced by the S-40542 treatment, while significantly decreased by bicalutamide. Oral

treatment with S-40542 resulted in a dose-dependent elevation of the plasma concentration, and its Cmax and AUC were much lower than those of bicalutamide. The pharmacokinetic study showed that this agent had relatively short plasma half-life and low oral bioavailability. Conclusion: S-40542 as well as bicalutamide

has shown as an anti-androgen by reducing the prostate weight of mice. Repeated oral treatment with S-40542 was shown to p53 inhibitor significantly suppress tumor growth in the KUCaP-2 xenograft model. “
“Objectives: We examined the effects of alpha1-adrenoceptor antagonist (tamsulosin hydrochloride) and antimuscarinic agent (solifenacin succinate) alone or in combination on the urinary adenosine triphosphate (ATP) level and cystometric parameters before Clomifene and after bladder stimulation. Methods: Female rats were administered tamsulosin hydrochloride (0.5 or 3 µg/kg/h) and/or solifenacin succinate (20 or 100 µg/kg/h) via a subcutaneously implanted osmotic minipump. Rats receiving distilled water were used as control. After 2 weeks, continuous cystometry with physiological saline or 0.1% acetic

acid solution was performed. Urinary ATP level was also measured before and after stimulation by 0.1% acetic acid solution. Results: During cystometry with bladder stimulation, the interval between voiding became shorter and the maximum voiding pressure (MVP) became higher in the control group. In the high-dose tamsulosin and solifenacin groups, the inhibition of urinary frequency was observed. The MVP also became higher in the high-dose tamsulosin group, but such a change was not seen in the high-dose solifenacin group. In case of low-dose administration, either agent alone did not inhibit the increase of urinary frequency and MVP due to bladder stimulation. However, co-administration of these ineffective low doses of tamsulosin and solifenacin resulted in the inhibition of urinary frequency. The high-dose or low-dose solifenacin group and the co-administration group showed similar inhibition of the increase of urinary ATP after bladder stimulation.

Plates blocked with PBS containing 10% FBS before 50 μL supernatants were added, and the incubated overnight at 4°C. After extensive washing, plates were incubated with a biotinylated anti-IFN-γ detection antibody. Plates were developed using avidin-peroxidase and 2-2′-azino-bis(3-ethyl-benzthiazoline-6-sulfonic acid) substrate (Sigma-Aldrich). OD405 was measured, and cytokine levels determined against a recombinant protein standard. All antibodies were purchased form BD Pharmingen. IFN-γ–producing cells were enumerated from splenocyte

CT99021 molecular weight populations isolated from immunized mice by cellular ELISPOT assay 47. Briefly, splenocytes (5×106 cells/mL) were cultured for 48 h in 24-well plates either with B5, B1 (10 μg/mL) or medium alone. Millititer HA nitrocellulose plates (Millipore) were coated overnight at 4°C with anti–IFN-γ. After blocking coated plates, antigen-stimulated cells were added at graded concentrations for 24 h at 37°C. learn more The wells were then incubated with biotin-conjugated anti–IFN-γmAb followed by incubation with avidin peroxidase (Vector Laboratories). Spots were developed by the addition of 3-amino-9-ethylcarbazole substrate (Sigma-Aldrich) and counted using a computerized image analysis system (Ligh-tools Research) and the image analyzer program, NIH Image 1.61. Immature BM-derived DC (1×106) were pulsed with 1×106 apoptotic (Ap-T) or untreated (T) T cells, peptide (20 μg/mL) or PBS for 8–12 h. In most experiments

DC were enriched by positive selection using anti-CD11c microbeads (Miltenyi) all and treated with activation modulators (for example, LPS (Sigma)) for 4–12 h CD11c+. DC were disabled (irradiated (3000 rad) or glutaraldehyde-fixed) and incubated with T responder cells (2×104/well) for the duration of the assay at 37°C in 5% CO2. For experiments analyzing the effect of antigen processing/presentation blockade, inhibitors

were first added to BM-derived DC for duration of 2 h – Concanamycin A (10–100 nM). DC were then washed and pulsed with peptide or apoptotic T cells for a total of 8 h (the final 4 h in the presence of LPS (1 μg/mL)). DC were positively selected using anti-CD11c microbeads (Miltenyi), and glutaraldehyde-fixed, before co-culturing with responder T cells. For IFN-γ secretion analysis, supernatants were harvested at 48 h. T-cell proliferation was measured by 3H-thymidine incorporation at 72 h. The desired number T cells were incubated in complete medium 4–12 h at 37°C in 5% CO2 in the presence of 5 μg/ml anti-Fas antibody (BD Pharmingen). To determine apoptosis induction 1×105 T cells in 100 μL buffer were stained with 10 μL/mL Annexin V FITC (BD Pharmingen). By flow cytometry apoptosis induction was confirmed using two parameters: (i) an anti-clockwise shift of the T-cell population in the forward versus side scatter dot plot, and (ii) a significant right shift of the peak on the FL1 histogram axis – indicating Annexin V staining.