Automated Cell Mechanical Characterization by On-Chip Sequential Squeezing: From Static to Dynamic.

The mechanical properties of cells are harmless biomarkers for cell identification and disease diagnosis. Although many systems have been developed to evaluate the static mechanical properties of cells for biomedical research, their robustness, effectiveness, and cost do not meet clinical requirements or the experiments with a large number of cell samples. In this paper, we propose an approach for on-chip cell mechanical characterization by analyzing the dynamic behavior of cells as they pass through multiple constrictions. The proposed serpentine microfluidic channel consisted of 20 constrictions connected in series and divided into five rows for tracking cell dynamic behavior. Assisted by computer vision, the squeezing time of each cell through five rows of constrictions was automatically collected and filtered to evaluate the cell's mechanical deformability. We observed a decreasing passage time and increasing dynamic deformability of the cells as they passed through the multiple constrictions. The deformability increase rate of the HeLa cells was eight times greater than that of MEF cells. Moreover, the weak correlation between the deformability increase rate and the cell size indicated that cell recognition based on measuring the deformability increase rate could hardly be affected by the cell size variation. These findings showed that the deformability increase rate of the cell under on-chip sequential squeezing as a new index has great potential in cancer cell recognition.

A Wearable Navigation Device for Visually Impaired People Based on the Real-Time Semantic Visual SLAM System.

Wearable auxiliary devices for visually impaired people are highly attractive research topics. Although many proposed wearable navigation devices can assist visually impaired people in obstacle avoidance and navigation, these devices cannot feedback detailed information about the obstacles or help the visually impaired understand the environment. In this paper, we proposed a wearable navigation device for the visually impaired by integrating the semantic visual SLAM (Simultaneous Localization And Mapping) and the newly launched powerful mobile computing platform. This system uses an Image-Depth (RGB-D) camera based on structured light as the sensor, as the control center. We also focused on the technology that combines SLAM technology with the extraction of semantic information from the environment. It ensures that the computing platform understands the surrounding environment in real-time and can feed it back to the visually impaired in the form of voice broadcast. Finally, we tested the performance of the proposed semantic visual SLAM system on this device. The results indicate that the system can run in real-time on a wearable navigation device with sufficient accuracy.

On-Chip Cell-Cell Interaction Monitoring at Single-Cell Level by Efficient Immobilization of Multiple Cells in Adjustable Quantities.

In recent decades, cell immobilization using microfluidic chips has facilitated significant advancements in biological analyses at the single-cell level. However, the efficient capture of multiple cells as a cluster in adjustable quantities for cell-cell interaction has not been achieved. In this paper, aiming to monitor the cell-cell interaction at the single-cell level, we proposed a novel method for the efficient immobilization of adjustable quantities of cells on the basis of passive hydrodynamics so that different cell-cell interaction patterns could be generated. Experiments were conducted to characterize the key geometric parameters of the chip to optimize the efficiency of trapping different quantities of cells. In the microfluidic chips optimized for immobilizing one to five cells, the trapping success rates (TSRs) were up to 97%, 87%, 84%, 58%, and 54%, respectively. Furthermore, the throughput was over 200 cells min-1 with a minimum cell density of 350 cells mm-2. Finally, in the experiments of applying the proposed multicell immobilization chips to cell-cell interaction monitoring, calcein-AM transfer between multiple cells under different patterns has been studied through quantifying the local fluorescent intensity. The results demonstrated that the proposed method could be a promising opportunity in the widening field of biological research at the single-cell level.

Prognostic impact of trisomy 21 in follicular lymphoma.

The chromosomal abnormalities associated with follicular lymphoma (FL) prognosis are not fully elucidated. Here, we evaluated the pattern of chromosomal abnormalities in FL, and clarified the correlations between the cytogenetic features and clinical outcome. Cytogenetic analysis was performed using standard methods of Giemsa-banding at diagnosis for 201 FL patients admitted to our hospitals between 2001 and 2013. The identified chromosomal abnormalities were: t(14;18)(q32;q21) (59·2%), +X (17·9%), del(6)(q)/-6 (16·9%), +7 (14·4%), abnormality of 1q12-21/1q (12·9%), del(13)(q)/-13 (11·9%), abnormality of 3q27 (10·4%), abnormality of 10q22-24 (10·0%), +12/dup(12)(q) (10·0%), abnormality of 1p21-22/1p (9·0%), +18 (9·0%), del(17)(p)/-17 (5·0%), and a complex karyotype (54·7%). Patients with trisomy 21 had a significantly shorter progression-free survival (P = 0·00171) and overall survival (OS) (P < 0·001) than those without trisomy 21; additionally, patients with trisomy 21 in the rituximab-treated cohort also had a significantly shorter OS (P = 0·000428). Multivariate analysis identified trisomy 21 as an independent risk factor in our cohorts with or without t(14;18) (P = 0·015). In conclusion, the presence of trisomy 21 was an independent risk factor for in FL. Chromosomal analysis of FL patients at diagnosis can provide useful information about their expected survival.

Multifunctional Noncontact Micromanipulation Using Whirling Flow Generated by Vibrating a Single Piezo Actuator.

A noncontact method that can achieve immobilization, transportation, and rotation in the microscale is desired in biological micromanipulation. A multifunctional noncontact micromanipulation method is proposed here based on a vibration-generated whirling flow. Resonance of a cantilever structure is utilized to extend the straight vibration of a single piezo actuator to the 2D circular vibration of a micropipette. The circular vibration in fluids can generate the whirling flow featured with low pressure in the core area and flow velocity gradient. The low pressure can immobilize the objects nearby and transport them together with the micropipette, and the flow velocity gradient is utilized to form a torque to rotate the immobilized object. Experiments of the microbeads are conducted to evaluate the claimed functions and quantify the key parameters that influence the rotation velocity. The cell spheroid is immobilized and rotated for 3D observation, and by assessing the viability of the cells containing in the spheroid, the proposed method is proved noninvasive to living cells. Finally, another important application in operations of mouse egg cells is shown, which indicates that the proposed method is a potential valuable tool in biological micromanipulation.

Hydrodynamic Tweezers: Trapping and Transportation in Microscale Using Vortex Induced by Oscillation of a Single Piezoelectric Actuator.

The demand for a harmless noncontact trapping and transportation method in manipulation and measurement of biological micro objects waits to be met. In this paper, a novel micromanipulation method named “Hydrodynamic Tweezers” using the vortex induced by oscillating a single piezoelectric actuator is introduced. The piezoelectric actuator is set between a micropipette and a copper beam. Oscillating the actuator at a certain frequency causes the resonance of the copper beam and extend 1D straight oscillation of the piezoelectric actuator to 2D circular oscillation of the micropipette, which induces a micro vortex after putting the micropipette into fluid. The induced vortex featuring low pressure in its core area can trap the object nearby. A robotic micromanipulator is utilized to transport the trapped objects together with the micropipette. Experiments of trapping and transportation microbeads are carried out to characterize the key parameters. The results show that the trapping force can be controlled by adjusting peak-peak voltage of the sinusoidal voltage input into the piezoelectric actuator.

Systemic lupus erythematosus as the concomitant manifestation of angioimmunoblastic T-cell lymphoma.

Herein we report a case of the simultaneous occurrence of angioimmunoblastic T-cell lymphoma (AITL) and systemic lupus erythematosus (SLE) in a 76-year-old woman. She presented with fever, night sweats, and general malaise. A laboratory examination revealed leukopenia, anemia, polyclonal hypergammaglobulinemia, hypocomplementemia, positive results for anti-nuclear antibodies and anti-double strand DNA (anti-dsDNA) antibodies, and mild proteinuria. A computed tomography scan of the abdominal cavity showed multiple swollen intra-abdominal and intra-pelvic lymph nodes. A biopsy specimen obtained from the peri-iliac lymph node confirmed the diagnosis of AITL, while renal biopsy results were consistent with lupus nephritis, International Society of Nephrology and Renal Pathology Society class V. These results indicated that our patient developed SLE concomitantly with AITL. These findings will lead to further understanding of the pathogenic mechanism of SLE.

A multicenter phase II prospective clinical trial of glucocorticoid for patients with untreated IgG4-related disease.

OBJECTIVE: Although glucocorticoids are effective for patients with IgG4-related disease, the treatment has not yet been standardized. Therefore, the treatment strategy should be established. PATIENTS AND METHODS: Patients who fulfilled the comprehensive diagnostic criteria for definite IgG4-related disease were started on prednisolone (0.6 mg/kg body weight) with the dose reduced every two weeks. The subsequent maintenance dose and need for prednisolone were determined for individual patients. The primary endpoint was the complete remission (CR) rate at one year. Secondary endpoints included overall response rate (ORR), the maintenance dose, the relapse rate, and adverse events. RESULTS: This study enrolled 61 patients. After clinicopathological review, three patients were excluded, and one, 13, and 44 patients were diagnosed with probable, possible, and definite IgG4-related disease, respectively. Of the 44 patients with definite IgG4-RD, 29 (65.9%) achieved CR, and the ORR was 93.2%. No patient was refractory to primary treatment. The most frequent adverse events were glucose intolerance. Six patients relapsed. CONCLUSIONS: Glucocorticoid treatment is usually effective for patients with IgG4-RD, and we should examine the possibility of other disorders when a patient is glucocorticoid refractory. Some patients are misdiagnosed, making central clinicopathological review of diagnosis very important in conducting clinical studies.

Multicentric Castleman's disease with multiple hepatic mass lesions mimicking malignant liver tumors.

Multicentric Castleman's disease (MCD) is a rare, non-malignant lymphoproliferative disorder. We report a case of MCD with multiple liver masses. A 26-year-old woman presented with asymptomatic anemia and hypoalbuminemia. Laboratory tests detected high CRP levels and findings indicative of polyclonal gammopathy. Abdominal CT revealed multiple hepatic large masses (≤10 cm) and partial calcification in the right lobe. Multiple enlarged lymph nodes were also identified in the cardiophrenic angle and porta hepatis. We suspected hepatic malignancy, but pathological examinations of the liver and lymph nodes demonstrated polyclonal plasma cell infiltration and fibrosis. IL-6 staining was positive for plasma cell infiltration of lymph nodes. A few plasma cells were positive for IgG4, and tests for HIV and HHV-8 were negative. Serum IL-6 and plasma VEGF levels were both elevated (45 and 536 pg/ml, respectively). The patient was diagnosed with plasma cell type MCD. We started treatment with PSL 1 mg/kg/day, which led to improvement of anemia, hypoalbuminemia, and high CRP levels. Marginal regression of liver masses was also observed. At the last follow-up, the patient had been progression-free for 18 months. To our knowledge, this is the first report of a plasma cell type MCD with liver masses.

Carcinosarcoma arising from uterine adenomyosis: A case report.

Carcinosarcoma arising from uterine adenomyosis is extremely rare. We encountered such a patient and herein provide a review of the literature. A 56-year-old woman was referred for a huge pelvic tumor, suspected to be an advanced uterine leiomyosarcoma. Intraoperative inspection revealed a mass, mainly located in the uterine myometrium, invading the uterine serosa. The tumor had previously spontaneously ruptured and disseminated to the pelvic cavity. Pathological and immunohistochemical examination revealed an infiltrative pattern of biphasic tumor cells composed of endometrioid adenocarcinoma and a nonepithelial component with rhabdomyosarcomatous differentiation. Benign endometrial glands with stromal cells were found adjacent to the area of the carcinosarcoma. The endometrium and both ovaries and fallopian tubes were microscopically free of tumor cells. The final diagnosis was heterologous type carcinosarcoma with rhabdomyosarcomatous differentiation, originating from uterine adenomyosis.

Proposed diagnostic criteria, disease severity classification, and treatment strategy for a novel disorder; TAFRO syndrome.

TAFRO syndrome is a systemic inflammatory disorder manifesting as thrombocytopenia; anasarca including pleural effusion and ascites; fever; renal insufficiency; and organomegaly including hepatosplenomegaly and lymphadenopathy. Its onset may be acute or sub-acute, but its etiology remains unknown. Although several clinical and pathological characteristics of TAFRO syndrome resemble those of Castleman's disease, other specific features can differentiate between the two. Some patients have been successfully treated with glucocorticoids and/or immunosuppressants including cyclosporin A, tocilizumab and rituximab, whereas others are refractory to treatment, eventually succumbing to the disease. Early and reliable diagnoses and early treatments with appropriate agents are essential to enhancing patient survival. The 2015 updated diagnostic criteria, disease severity classification and treatment strategy for TAFRO syndrome, as formulated by Japanese research teams, are presented herein. Furthermore, clinicopathological data on 28 patients with this condition and similar symptoms (e.g., MCD with serositis and thrombocytopenia) were analyzed retrospectively.

[Monomorphic post-transplant T-lymphoproliferative disorder after autologous stem cell transplantation for multiple myeloma].

We report a rare case of T cell type monomorphic post-transplant lymphoproliferative disorders (PTLD) after autologous stem cell transplantation. A 53-year-old man with multiple myeloma received autologous stem cell transplantation and achieved a very good partial response. Nine months later, he developed a high fever and consciousness disturbance, and had multiple swollen lymph nodes and a high titer of Epstein-Barr (EB) virus DNA in his peripheral blood. Neither CT nor MRI of the brain revealed any abnormalities. Cerebrospinal fluid contained no malignant cells, but the EB virus DNA titer was high. Lymph node biopsy revealed T cell type monomorphic PTLD. Soon after high-dose treatment with methotrexate and cytosine arabinoside, the high fever and consciousness disturbance subsided, and the lymph node swelling and EB virus DNA disappeared. Given the efficacy of chemotherapy in this case, we concluded that the consciousness disturbance had been induced by central nervous system involvement of monomorphic PTLD.

Piezo-actuated parallel mechanism for biological cell release at high speed.

In this paper, a dynamic releasing approach is proposed for high-speed biological cell manipulation. A compact parallel mechanism for grasping and releasing microobjects is used to generate controllable vibration to overcome the strong adhesion forces between the end effector and the manipulated object. To reach the required acceleration of the end effector, which is necessary for the detachment of the target object by overcoming adhesion forces, vibration in the end effector is generated by applying sinusoidal voltage to the PZT actuator of the parallel mechanism. For the necessary acceleration, we focus on the possible range of the frequency of the PZT-actuator-induced vibration, while minimizing the amplitude of the vibration (14 nm) to achieve precise positioning. The effect of the air and liquid environments on the required vibration frequency for successful release is investigated. For the first time, release results of microbeads and biological cells are compared. Release of the biological cells with 100 % success rate suggests that the proposed active release method is an appropriate solution for adhered biological cells during the release task.

Aspartate aminotransferase-to-platelet ratio index is associated with liver cirrhosis in patients undergoing surgery for hepatocellular carcinoma.

BACKGROUND: Among various preoperative evaluations of liver function, accurate assessment of liver cirrhosis (LC) is especially important in patients undergoing surgery for hepatocellular carcinoma (HCC). OBJECTIVE: To explore the most significant laboratory parameter associated with LC in patients undergoing surgery for HCC. METHODS: From among 588 HCC patients in our collected database who underwent liver surgery, 371 for whom sufficient laboratory data were evaluable, including direct serum fibrosis markers such as hyaluronic acid and type 3 procollagen peptide (P-3-P), were enrolled. Receiver operating characteristic (ROC) curve analysis was used to define the ideal cutoff values of laboratory parameters, and the area under the ROC curve for LC was measured. Univariate and multivariate analyses were performed to clarify the laboratory parameter most significantly associated with LC. RESULTS: Multivariate analysis of 13 laboratory parameters that had been selected by univariate analysis showed that the aspartate aminotransferase-to-platelet ratio index (APRI) (≤ 0.8/>0.8) (odds ratio, 2.687; 95% confidence interval 1.215-5.940; P = 0.015) was associated with LC, along with the aspartate aminotransferase to alanine aminotransferase ratio, the indocyanine green retention ratio at 15 min (ICG R15), and the level of hyaluronic acid. Among these four parameters associated with LC, ROC curve analysis revealed that APRI (0.757) had the largest area under the ROC (aspartate aminotransferase to alanine aminotransferase 0.505, ICG R15 0.714, and hyaluronic acid 0.743). CONCLUSIONS: APRI is closely associated with LC in patients undergoing surgery for HCC.

Cytological findings for the diagnosis of primary thyroid mucosa-associated lymphoid tissue lymphoma by fine needle aspiration.

OBJECTIVE: We examined cytological findings for the diagnosis of primary thyroid mucosa-associated lymphoid tissue (MALT) lymphoma by fine needle aspiration. STUDY DESIGN: During the study period of 4 years, a total of 101 cases including 51 MALT lymphomas, 20 Hashimoto's thyroiditis (HT), and 30 diffuse large-cell B-cell lymphomas were cytologically examined. MALT lymphomas were divided into 44 common MALT and 7 MALT lymphomas with extreme plasmacytic differentiation (MALT-EPCD). RESULTS: (1) Small- to medium-sized cells displaying irregularly shaped nuclei with prominent nucleoli (ISN-PN) were neoplastic cells. (2) In the case of a frequency of plasma cells (PC) below 15%, the accuracy rate for distinguishing common MALT from HT was 97% for ISN-PN cell frequencies above 20% in combination with the presence of lymphoepithelial lesion clusters (LELC) and mountain range-like clusters (MRLC). The frequency of large-sized cells was below 15% in common MALT. (3) In the case of a frequency of PC above 15%, cases with a sum of PC and ISN-PN cells above 30% were MALT-EPCD. (4) MRLC were cell clusters derived from regions of follicular colonization, and LELC were cell clusters from lymphoepithelial lesions of MALT lymphomas. CONCLUSION: Useful cytological criteria for the diagnosis of thyroid primary MALT lymphoma, such as neoplastic cells and cell clusters, were defined.

[Diffuse large B-cell lymphoma of the testis relapsed 16 years after achieving complete response].

Testicular lymphoma is a rare disease, accounting for 1-2% of non-Hodgkin lymphoma and 5-9% of all testicular tumors, and has a high relapse rate with a poor prognosis. We report a patient with testicular diffuse large B-cell lymphoma (DLBCL) who relapsed after being in remission for 16 years. He had undergone orchiectomy of the right testis and was diagnosed as having DLBCL (stage IAE) at 49 years of age. After 3 cycles of CHOP, he achieved a complete remission. Orchiectomy was performed because of a left testicular tumor, and he was again diagnosed with DLBCL at the age of 65. VH3-21 was detected in lymphoma cells at the times of both the first diagnosis and the relapse based on analysis of the variable region of the immunoglobulin heavy chain. Accordingly, the lymphoma cells at relapse were confirmed to be the same clone as that which had been documented at the first diagnosis.

The first report of adolescent TAFRO syndrome, a unique clinicopathologic variant of multicentric Castleman's disease.

BACKGROUND: TAFRO syndrome is a unique clinicopathologic variant of multicentric Castleman's disease that has recently been identified in Japan. It is characterized by a constellation of symptoms: Thrombocytopenia, Anasarca, reticulin Fibrosis of the bone marrow, Renal dysfunction and Organomegaly (TAFRO). Previous reports have shown that affected patients usually respond to immunosuppressive therapy, but the disease sometimes has a fatal course. TAFRO syndrome occurs in the middle-aged and elderly and there are no prior reports of the disease in adolescents. Here we report the first adolescent case, successfully treated with anti-IL-6 receptor antibody (tocilizumab, TCZ) and monitored with serial cytokine profiles. CASE PRESENTATION: A 15-year-old Japanese boy was referred to us with fever of unknown origin. Whole body computed tomography demonstrated systemic lymphadenopathy, organomegaly and anasarca. Laboratory tests showed elevated C-reactive protein and hypoproteinemia. Bone marrow biopsy revealed a hyperplastic marrow with megakaryocytic hyperplasia and mild reticulin fibrosis. Despite methylprednisolone pulse therapy, the disease progressed markedly to respiratory distress, acute renal failure, anemia and thrombocytopenia. Serum and plasma levels of cytokines, including IL-6, vascular endothelial growth factor, neopterin and soluble tumor necrosis factor receptors I and II, were markedly elevated. Repeated weekly TCZ administration dramatically improved the patient's symptoms and laboratory tests showed decreasing cytokine levels. CONCLUSION: To our knowledge, this is the first report of TAFRO syndrome in a young patient, suggesting that this disease can occur even in adolescence. The patient was successfully treated with TCZ. During our patient's clinical course, monitoring cytokine profiles was useful to assess the disease activity of TAFRO syndrome.

Micromixer driven by bubble-induced acoustic microstreaming for multi-ink 3D bioprinting.

Recently, the 3D printing of cell-laden hydrogel structures, known as bioprinting, has received increasing attention owing to advances in tissue engineering and drug screening. However, a micromixing technology that efficiently mixes viscous bioinks under mild conditions is needed. Therefore, this study presents a novel method for achieving homogeneous mixing of multiple inks in 3D bioprinting through acoustic stimulation. This technique involves generating an acoustic microstream through bubble oscillations inside a 3D bioprinting nozzle. We determined the optimal hole design for trapping a bubble, hole arrangement, and voltage for efficient mixing, resulting in a four-fold increase in mixing efficiency compared to a single bubble arrangement. Subsequently, we propose a nozzle design for efficient mixing during bioprinting. The proposed nozzle design enabled the successful printing of line structures with a uniform mixture of different viscous bioinks, achieving a mixing efficiency of over 80% for mixing 0.5-1.0 wt% sodium alginate aqueous solutions. Additionally, acoustic stimulation had no adverse effects on cell viability, maintaining a high cell viability of 88% after extrusion. This study presents the first use of a bubble micromixer in 3D bioprinting, demonstrating gentle yet effective multi-ink mixing. We believe this approach will broaden 3D printing applications, particularly for constructing functional structures in 3D bioprinting.

Characterization of Chitosan Hydrogels Obtained through Phenol and Tripolyphosphate Anionic Crosslinking.

Chitosan is a deacetylated polymer of chitin that is extracted mainly from the exoskeleton of crustaceans and is the second-most abundant polymer in nature. Chitosan hydrogels are preferred for a variety of applications in bio-related fields due to their functional properties, such as antimicrobial activity and wound healing effects; however, the existing hydrogelation methods require toxic reagents and exhibit slow gelation times, which limit their application in biological fields. Therefore, a mild and rapid gelation method is necessary. We previously demonstrated that the visible light-induced gelation of chitosan obtained through phenol crosslinking (ChPh) is a rapid gelation method. To further advance this method (<10 s), we propose a dual-crosslinked chitosan hydrogel obtained by crosslinking phenol groups and crosslinking sodium tripolyphosphate (TPP) and the amino groups of chitosan. The chitosan hydrogel was prepared by immersing the ChPh hydrogel in a TPP solution after phenol crosslinking via exposure to visible light. The physicochemical properties of the dual-crosslinked hydrogels, including Young's moduli and water retentions, were subsequently investigated. Young's moduli of the dual-crosslinked hydrogels were 20 times higher than those of the hydrogels without TPP ion crosslinking. The stiffness could be manipulated by varying the immersion time, and the water retention properties of the ChPh hydrogel were improved by TPP crosslinking. Ion crosslinking could be reversed using an iron chloride solution. This method facilitates chitosan hydrogel use for various applications, particularly tissue engineering and drug delivery.