Neutrophil-to-lymphocyte ratio associates with nutritional parameters, intratumoral immune profiles, and clinical outcomes of pembrolizumab in head and neck squamous cell carcinoma.

BACKGROUND: The relationship between the tumor-immune microenvironment and systemic inflammatory markers, including neutrophil-to-lymphocyte ratio (NLR), is unclear. METHODS: We examined the characteristics of systemic inflammatory markers and tumor immune microenvironments in relation to treatment outcomes in 29 consecutive patients with recurrent/metastatic head and neck squamous cell carcinoma (R/M HNSCC) who received pembrolizumab, using 14-marker multiplex immunohistochemistry and image cytometry. RESULTS: NLR ≥4.5 (high NLR) at pretreatment status significantly correlated with short overall survival (OS) and progression-free survival-2 (PFS2) and malnutrition status. High NLR in peripheral blood was significantly correlated with low lymphoid cell and high tumor-associated macrophage counts in tissues, especially myeloid-to-lymphoid cell ratios, suggesting an association between circulating and intratumoral immune complexity profiles. CONCLUSIONS: This study suggests a link between NLR in circulating blood, systemic nutritional status, and immune composition within the tumor.

A rare case of positional vertebrobasilar ischemia with the retrograde flow of the vertebral artery.

Positional vertebrobasilar ischemia, also known as Bow hunter stroke, is typically caused by mechanical compression of the vertebral artery (VA). On the other hand, subclavian steal syndrome is incidentally detected by vertigo, syncope or loss of consciousness due to the steal phenomenon. A 61-year-old man suffered from near syncope when he turned his head to the left side. Although asymmetric arm blood pressure of the right dominant was observed, arm claudication was not detected. Computed tomographic angiography and magnetic resonance imaging revealed total occlusion of the left subclavian artery, hypoplasia of the right VA and an incomplete circle of Willis. Furthermore, carotid Doppler ultrasonic echography revealed retrograde blood flow of the left VA. It means that head rotation might occur in the left VA ischemia. An axillary-axillary bypass surgery was performed and ultrasonic echography showed anterograde effective blood flow of the left VA after the procedure.

Spatially resolved immune microenvironmental profiling for follicular thyroid carcinoma with minimal capsular invasion.

Spatial profiles of the tumor-immune microenvironment are associated with disease progression and clinicopathological factors in various cancers. Follicular thyroid carcinoma (FTC) is the second most common thyroid cancer, where the presence of capsular invasion or angioinvasion determines the pathological diagnosis; however, little is known about the immune microenvironment profiles associated with the acquisition of invasive potential of FTC. In this study, we focused on FTC with minimal capsular invasion, and the spatially resolved immune microenvironment of FTC was studied in the discovery (n = 13) and validation cohorts (n = 40). CD8+ T cells, helper T cells, regulatory T cells, B cells, natural killer cells, tumor-associated macrophages, CD66+ granulocytes, mature dendritic cells, and mast cells were quantitatively evaluated in single tissue sections, via a 12-marker multiplex immunohistochemistry and image cytometry. Cell densities and compositions of immune cells were spatially stratified by six tissue regions including tumor center, subcapsular region, capsular invasion, adjacent stroma of capsular invasion, peritumoral stroma, and adjacent normal. Lymphoid cell lineages in the tumor center and subcapsular regions were significantly lower than those in adjacent normal and peritumoral stroma, potentially related to the lymphoid lineage exclusion from the intratumoral regions of FTC. Interestingly, immune cell composition profiles in the capsular invasive front were distinct from those of intratumoral region. The ratios of T cells to CD66b+ granulocytes with capsular invasion were significantly higher than those without capsular invasion, suggesting the presence of a unique immune microenvironment at the invasive front between tumor foci and stroma. In addition, tumor cells at the capsular invasive front showed significantly higher expression of tumor programmed cell death ligand 1 (PD-L1) than those at the tumor center. This study revealed spatial immune profiles associated with capsular invasion of FTC, providing new insights into the mechanisms underlying its development and initial invasion.

Elevation of circulating neutrophil extracellular traps, interleukin (IL)-8, IL-22, and vascular endothelial growth factor in patients with venomous snake mamushi (Gloydius blomhoffii) bites.

Mamushi bites cause swelling and pain that extend from the bitten site. The coagulopathic, anti-coagulopathic, and vasculopathic actions of mamushi venom result in various laboratory abnormalities, occasionally with muscular, renal, and other organ damage. We investigated the serum biomarkers that were associated with the pathogenesis of mamushi bites, focusing on markers related to tissue-damage and neutrophil activation. Twenty patients (one case of grade 2, 13 cases of grade 3, and six cases of grade 4 of severity) seen by us in one summer season were enrolled. Peripheral blood samples were taken from the patients on day 0, day 2, and day 7 after mamushi bites. In addition to routine blood examination, serum samples were subjected to enzyme-linked immunosorbent assay for citrullinated histone H3 (CitH3), interleukin (IL)-8, IL-17A, IL-22, vascular endothelial growth factor (VEGF), high mobility group box protein 1 (HMGB1), tumor necrosis factor (TNF)-α, and IL-33. Creatinine kinase (CK) values significantly correlated with prothrombin time (PT) levels, suggesting that muscular damage is associated with exaggerated coagulation and fibrinolysis. In the vast majority of patients, HMGB1, TNF-α, and IL-33 were under detection levels. Neutrophil counts did not correlate with PT or CK, indicating that the coagulation disorder and muscular damage were virtually independent of the neutrophil activation. The neutrophil number significantly correlated with CitH3, a representative marker of neutrophil extracellular traps. Moreover, there were significant correlations between neutrophil number, CitH3, IL-8, IL-22, and VEGF. Our study suggests that there are two major cascades in mamushi bites. One is an already characterized venom effect on coagulation, vessels, and muscles. In the other novel cascade, we propose that neutrophil activation with IL-8 leads to the production of IL-22 and VEGF. This sequential event may contribute to both vascular damage and repair.

Questionnaire Survey of Possible Association of Allergic Diseases with Adverse Reactions to SARS-CoV-2 Vaccination.

To protect against COVID-19, SARS-CoV-2 vaccines have been widely used. Besides anaphylaxis, some less severe adverse effects may occur at higher frequencies. It remains unclear whether present or past histories of allergic diseases exert effects on local and systemic reactions. We conducted a questionnaire survey among workers in our hospital. We analyzed the adverse effects occurring after the first and second doses of the Pfizer-BioNTech vaccine in 955 subjects. The presence or absence of local injection reactions and systemic reactions (headache, fatigability, fever, muscle pain, and joint pain) was questioned. The intensities of these reactions were graded on a scale of 0-4 (except fever) or 0-2 (fever). The allergic diseases that we focused on were bronchial asthma, atopic dermatitis, food allergy, pollinosis, and hand eczema. For the systemic reactions, fatigability after the first dose tended to be more severe in the bronchial asthma than in the non-allergic group. Headache, joint pain, and fever tended to be more severe in the food allergy than in the non-allergic group after the second dose. For the local skin reactions, atopic dermatitis subjects tended to show rather less severe local skin reactions after the second dose. The results contribute to the guidelines for the care of individuals with different allergy histories, so that they may safely receive their vaccine.

[Corrigendum] Loss of Runt­related transcription factor 3 induces resistance to 5­fluorouracil and cisplatin in hepatocellular carcinoma.

Following the publication of this article, we realize that there were some errors in the manuscript. Details of the experiments describing the gene silencing of RUNX3 with small interfering RNA (siRNA) were erroneously included in this paper, and all references to siRNA should have been deleted from the manuscript prior to publication. In the subsection entitled 'Cell lines and cell culture' on page 2577, the left­hand column, the text should have indicated that the human HCC cell lines Hep3B and Huh7 were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA), whereas HLF cells were obtained from the Japanese Cancer Resources Bank (Tokyo, Japan). Lastly, an error was made in describing the calculation of the IC50 values, which did not correlate with the data shown in Fig. 2. Therefore, the subsection entitled 'Ectopic RUNX3 protein expression suppresses cell growth...' should have been entitled 'Ectopic RUNX3 protein expression increases 5­FU and CDDP sensitivity', and the text herein should have read as follows: We analyzed the effects of RUNX3 on chemosensitivity in the RUNX3­ or CAT (mock)­transfected Hep3B and Huh7 cells. RUNX3 expression enhanced 5­FU sensitivity in both cell lines; the cell viability with 5­FU (100 nM) decreased from 66.3±4.6 to 34.3±5.0%, and from 71.0±4.7% to 27.0±5.5% in the Hep3B and Huh7 cells, respectively (Fig. 2A). RUNX3 expression also enhanced CDDP sensitivity in both cell lines; the cell viability with CDDP (100 nM) decreased from 58.7±2.6% to 25.7±4.9%, and from 67.7±4.1% to 25.7±7.5% in the Hep3B and Huh7 cells, respectively (Fig. 2B). We sincerely apologize for these errors and oversights, which have not affected any of the overall conclusions reported in the study, and regret any inconvenience they may have caused. [the original article was published in the Oncology Reports 35: 2576-2582, 2016; DOI: 10.3892/or.2016.4681].

Dynamic Determinants of the Uncontrolled Manifold during Human Quiet Stance.

Human postural sway during stance arises from coordinated multi-joint movements. Thus, a sway trajectory represented by a time-varying postural vector in the multiple-joint-angle-space tends to be constrained to a low-dimensional subspace. It has been proposed that the subspace corresponds to a manifold defined by a kinematic constraint, such that the position of the center of mass (CoM) of the whole body is constant in time, referred to as the kinematic uncontrolled manifold (kinematic-UCM). A control strategy related to this hypothesis (CoM-control-strategy) claims that the central nervous system (CNS) aims to keep the posture close to the kinematic-UCM using a continuous feedback controller, leading to sway patterns that mostly occur within the kinematic-UCM, where no corrective control is exerted. An alternative strategy proposed by the authors (intermittent control-strategy) claims that the CNS stabilizes posture by intermittently suspending the active feedback controller, in such a way to allow the CNS to exploit a stable manifold of the saddle-type upright equilibrium in the state-space of the system, referred to as the dynamic-UCM, when the state point is on or near the manifold. Although the mathematical definitions of the kinematic- and dynamic-UCM are completely different, both UCMs play similar roles in the stabilization of multi-joint upright posture. The purpose of this study was to compare the dynamic performance of the two control strategies. In particular, we considered a double-inverted-pendulum-model of postural control, and analyzed the two UCMs defined above. We first showed that the geometric configurations of the two UCMs are almost identical. We then investigated whether the UCM-component of experimental sway could be considered as passive dynamics with no active control, and showed that such UCM-component mainly consists of high frequency oscillations above 1 Hz, corresponding to anti-phase coordination between the ankle and hip. We also showed that this result can be better characterized by an eigenfrequency associated with the dynamic-UCM. In summary, our analysis highlights the close relationship between the two control strategies, namely their ability to simultaneously establish small CoM variations and postural stability, but also make it clear that the intermittent control hypothesis better explains the spectral characteristics of sway.

Loss of Runt-related transcription factor 3 induces resistance to 5-fluorouracil and cisplatin in hepatocellular carcinoma.

Runt-related transcription factor 3 (RUNX3) is known to function as a tumor suppressor in gastric cancer and other types of cancers, including hepatocellular carcinoma (HCC). However, its role has not been fully elucidated. In the present study, we aimed to evaluate the role of RUNX3 in HCC. We used the human HCC cell lines Hep3B, Huh7 and HLF; RUNX3 cDNA was introduced into Hep3B and Huh7 cells, which were negative for endogenous RUNX3 expression, and RUNX3 siRNA was transfected into HLF cells, which were positive for endogenous RUNX3. We analyzed the expression of RUNX3 and multidrug resistance-associated protein (MRP) by immunoblotting. MTT assays were used to determine the effects of RUNX3 expression on 5-fluorouracil (5-FU) and cisplatin (CDDP) sensitivity. Finally, 23 HCC specimens resected from patients with HCC at Okayama University Hospital were analyzed, and correlations among immunohistochemical expression of RUNX3 protein and MRP protein were evaluated in these specimens. Exogenous RUNX3 expression reduced the expression of MRP1, MRP2, MRP3 and MRP5 in the RUNX3-negative cells, whereas knockdown of RUNX3 in the HLF cells stimulated the expression of these MRPs. An inverse correlation between RUNX3 and MRP expression was observed in the HCC tissues. Importantly, loss of RUNX3 expression contributed to 5-FU and CDDP resistance by inducing MRP expression. These data have important implications in the study of chemotherapy resistance in HCC.

Non-actively controlled double-inverted-pendulum-like dynamics can minimize center of mass acceleration during human quiet standing.

Multiple joint movements during human quiet standing exhibit characteristic inter-joint coordination, shortly referred to as reciprocal relationship, in which angular acceleration of the hip joint is linearly and negatively correlated with that of the ankle joint (antiphase coordination) and, moreover, acceleration of the center of mass (CoM) of the double-inverted-pendulum (DIP) model of the human body is close to zero constantly. A question considered in this study is whether the reciprocal relationship is established by active neural control of the posture, or rather it is a biomechanical consequence of non-actively controlled body dynamics. To answer this question, we consider a DIP model of quiet standing, and show that the reciprocal relationship always holds by Newton's second law applied to the DIP model with human anthropometric dimensions, regardless of passive and active joint torque patterns acting on the ankle and hip joints. We then show that characteristic frequencies included in experimental sway trajectories with the reciprocal relationship match with harmonics of the eigenfrequency of the stable antiphase eigenmode of the non-actively controlled DIP-like unstable body dynamics. The results suggest that non-actively controlled DIP-like mechanical dynamics is a major cause of the minimization of the CoM acceleration during quiet standing, which is consistent with a type of control strategy that allows switching off active neural control intermittently for suitable periods of time during quiet standing.

Functional dissociation in right inferior frontal cortex during performance of go/no-go task.

The contribution of the right inferior frontal cortex to response inhibition has been demonstrated by previous studies of neuropsychology, electrophysiology, and neuroimaging. The inferior frontal cortex is also known to be activated during processing of infrequent stimuli such as stimulus-driven attention. Response inhibition has most often been investigated using the go/no-go task, and the no-go trials are usually given infrequently to enhance prepotent response tendency. Thus, it has not been clarified whether the inferior frontal activation during the go/no-go task is associated with response inhibition or processing of infrequent stimuli. In the present functional magnetic resonance imaging study, we employed not only frequent-go trials but also infrequent-go trials that were presented as infrequently as the no-go trials. The imaging results demonstrated that the posterior inferior frontal gyrus (pIFG) was activated during response inhibition as revealed by the no-go vs. infrequent-go trials, whereas the inferior frontal junction (IFJ) region was activated primarily during processing of infrequent stimuli as revealed by the infrequent-go versus frequent-go trials. These results indicate that the pIFG and IFJ within the inferior frontal cortex are spatially close but are associated with different cognitive control processes in the go/no-go paradigm.

Differential superior prefrontal activity on initial versus subsequent shifts in naive subjects.

Flexible adaptation to changing environments requires shifting of a cognitive set, one basic function of the prefrontal cortex. Set shifting, as instantiated in the Wisconsin Card Sorting Task (WCST) administered in a neuropsychological testing room, is typically achieved when subjects have no prior experiences of updating one WCST behavior to another. By contrast, earlier neuroimaging studies typically involved examination of repeated transitions between particular behaviors, to which situation subjects are far from naive. Naive subjects with no prior knowledge of the WCST were recruited in the present functional magnetic resonance imaging study to test set shifting under unknown situations that they experienced for the first time. Prominent activation was revealed in the left superior prefrontal cortex selectively on the initial shifts. On the other hand, the inferior prefrontal cortex was significantly activated on both the initial and subsequent shifts. The superior prefrontal activation distinguishable from the conventional inferior prefrontal activation suggests a selective role of this region in performance of the WCST in naive subjects.

On verbal/nonverbal modality dependence of left and right inferior prefrontal activation during performance of flanker interference task.

One of the most prevailing views on the functional localization of human cognition is the hemispheric specialization, wherein the left and right hemispheres are implicated primarily in verbal and nonverbal functions, respectively. Cognitive control is known to involve the lateral prefrontal cortex. However, it remains unclear whether the hemispheric specialization in the lateral prefrontal cortex can be observed in cognitive control per se, independent of sensory aspects of stimulus materials. In this functional magnetic resonance imaging study, we tested whether the verbal/nonverbal hemispheric specialization applies to the lateral prefrontal activation by investigating interference suppression, the ability to filter out irrelevant information in the environment. The flanker task was employed using a compound stimulus that contained a target and a flanker. The flanked stimulus was either a color word flanked by a colored patch or a colored patch flanked by a color word, which allowed us to manipulate the modality of the presented flanker stimulus from which interference originates, keeping the total stimulus modality balanced. The inferior frontal gyrus (IFG) showed prominent Modality-by-Hemisphere interaction in interference suppression, the left IFG being activated when a word flanker (plus a patch target) was presented and the right IFG being activated when a patch flanker (plus a word target) was presented. These results suggest that the verbal/nonverbal hemispheric specialization in the IFG can be explained by cognitive control processes per se, independent of sensory aspects of presented materials.

Primary effect of bromoxynil to induce plant cell death may be cytosol acidification.

Bromoxynil, 3,5-dibromo-4-hydroxybenzonitrile, is a commonly used herbicide and is also used as a tool to trigger rapid cell death in basic botany. However, the primary effect inducing cell death is not known. Bromoxynil inhibited the cytoplasmic streaming and killed cells in Chara corallina when it was applied in the acidic external medium. At higher pH, bromoxynil was inert even at high concentrations. It was speculated that bromoxynil in the protonated form enters the cell and acidifies the cytosol by releasing H(+). Experiments using analogues of bromoxynil supported this possibility. Acidification of the cytosol by bromoxynil was confirmed by experiments using pollen tubes. Based on the acidity of the apoplast, the herbicide action of bromoxynil in higher plants was discussed.