Study on degranulation of mast cells under C48/80 treatment by electroporation-assisted and ultrasound-assisted surface-enhanced Raman spectrascopy.

Both electroporation-assisted and ultrasound-assisted delivery methods can rapidly deliver nanoparticles into living cells for surface-enhanced Raman scattering (SERS) detection, but these two methods have never been compared. In this study, electroporation-assisted SERS and ultrasound-assisted SERS were employed to detect the biochemical changes of degranulated mast cells induced by mast cell stimulator (C48/80). The results showed that the cell damage of electroporation based on controllable electric pulse was smaller than that of ultrasound based on cavitation. Transmission electron microscope images of cells indicated that the nanoparticles delivered by electroporation were mainly distributed in the cytoplasm, while ultrasound could transport nanoparticles to the cytoplasm and nucleus. Therefore, electroporation-assisted SERS mainly detects the biochemical information of cytoplasm, while ultrasound-assisted SERS gets more spectral signals of nucleic acid. Both methods can obtain high quality SERS signal of cells. With drug treatment, the SERS peak intensity of 733 cm-1 attributed to phosphatidylserine decreased significantly, which may be due to the activation of mast cell degranulation pathway stimulated by C48/80 agonist, resulting in a large amount of intracellular serine being used to synthesize tryptase, while the production of phosphatidylserine decreased. Further, based on principal component analysis and linear discriminant analysis (PCA-LDA approach), ultrasound-assisted SERS could achieve better sensitivity, specificity and accuracy in the discrimination and identification of drug-treated degranulated mast cells than electroporation assisted SERS. This exploratory work is helpful to realize the real-time dynamic SERS detection of intracellular biochemical components, and it also has great potential in intracellular SERS analysis, such as the cytotoxicity assay of anti-tumor drugs or cancer cell screening.

Label-free surface enhanced Raman spectroscopy analysis of blood serum via coffee ring effect for accurate diagnosis of cancers.

Surface-enhanced Raman spectroscopy (SERS) is considered as an ultrasensitive, non-invasive as well as rapid detection technology for cancer diagnosis. In this study, we developed a novel blood serum analysis strategy using coffee ring effect-assisted label-free SERS for different types of cancer screening. Additionally, the pretreated Ag nanoparticles (Ag NPs) were mixed with the serum from liver cancer patients (n = 40), prostate cancer patients (n = 32) and healthy volunteers (n = 30) for SERS measurement. The droplets of Ag NPs-serum mixture formed the coffee ring on the peripheral after air-drying, and thus extremely enhancing Raman signal and ensuring the stability and reliability of SERS detection. Partial least square (PLS) and support vector machine (SVM) algorithms were utilized to establish the diagnosis model for SERS spectra data classifying, yielding the high diagnostic accuracy of 98.04% for normal group and two types of cancers simultaneously distinguishing. More importantly, for the unknown testing set, an ideal diagnostic accuracy of 100% could be achieved by PLS-SVM algorithm for differentiating cancers from the normal group. The results from this exploratory work demonstrate that serum SERS detection combined with PLS-SVM diagnostic algorithm and coffee ring effect has great potential for the noninvasive and label-free detection of cancer.

Quantitative detection of crystal violet using a surface-enhanced Raman scattering based on a flower-like HAp/Ag nanocomposite.

Herein, we proposed a simple one-pot sol-thermal strategy to prepare a highly sensitive and reproducible SERS substrate. The silver-doped hydroxyapatite nanocomposite (HAp/Ag) could suppress the oxidation of silver nanoparticles, which endow the SERS substrate with good stability and reproducibility. Due to the strong interaction between the HAp/Ag substrate and the analytes, a stronger Raman signal generated during the process of SERS detection. In particular, the HAp/Ag substrate enabled the determination of rhodamine 6G (R6G) and crystal violet (CV), and the limits of detection (LOD) were low at 10-6 M and 10-5 M, respectively. In addition, the HAp/Ag substrate could be used for the quantitative analysis of CV in wastewater with a good linear relationship between 10-2 and 10-5 M. In this context, the HAp/Ag substrate combines the superior properties of both Ag NPs and HAp particles, providing a potential method for monitoring the environment and building a convenient SERS platform to detect pollutants in wastewater.

Label-free determination of liver cancer stages using surface-enhanced Raman scattering coupled with preferential adsorption of hydroxyapatite microspheres.

Here, we explored a label-free albumin targeted analysis method by utilizing hydroxyapatite (HAp) to adsorb-release serum albumin, in conjunction with surface-enhanced Raman scattering (SERS) for screening liver cancer (LC) at different tumor (T) stages. Excitingly, albumin can be preferentially adsorbed by HAp as compared with other serum proteins. Moreover, we developed a novel strategy using a high concentration of PO43- solution as the albumin-release agent. This method overcomes the shortcomings of the traditional purification technology of serum albumin, which requires acid to release protein, and ensures that the structure and properties of albumin are not damaged. The SERS spectra of serum albumin obtained from three sample groups were analyzed to verify the feasibility of this new method: healthy volunteers (n = 35), LC patients with T1 stage (n = 25) and LC patients with T2-T4 stage (n = 23). Furthermore, principal component analysis (PCA) combined with linear discriminant analysis (LDA) was employed to classify the early T (T1) stage LC vs. normal group and advanced T (T2-T4) stage LC vs. normal group, yielding high diagnostic accuracies of 90.00% and 96.55%, respectively, which showed a 10% improvement in diagnostic accuracy for the early stage detection of cancer as compared with previous studies. The results of this exploratory work demonstrated that HAp-adsorbed-released serum albumin combined with SERS analysis has great potential for label-free, noninvasive and sensitive detection of different T stages of liver cancer.

Label-free diagnosis of breast cancer based on serum protein purification assisted surface-enhanced Raman spectroscopy.

Serum protein is generally used to assess the severity of disease, as well as cancer progression and prognosis. Herein, a simple and rapid serum proteins analysis method combined with surface-enhanced Raman spectroscopy (SERS) technology was applied for breast cancer detection. The cellulose acetate membrane (CA) was employed to extract human serum proteins from 30 breast cancer patients and 45 healthy volunteers and then extracted proteins were mixed with silver nanoparticles for SERS measurement. Additionally, we also mainly assessed the use of different ratios of proteins-silver nanoparticles (Ag NPs) mixture to generate maximum SERS signal for clinical samples detection. Two multivariate statistical analyses, principal component analysis-linear discriminate analysis (PCA-LDA) and partial least square-support vector machines (PLS-SVM) were used to analyze the obtained serum protein SERS spectra and establish the diagnostic model. The results demonstrate that the PLS-SVM model provides superior performance in the classification of breast cancer diagnosis compared with PCA-LDA. This exploratory work demonstrates that the label-free SERS analysis technique combined with CA membrane purified serum proteins has great potential for breast cancer diagnosis.

High spatio-temporal resolution measurement of A1 R and A2A R interactions combined with Iem-spFRET and E-FRET methods.

A1 R-A2A R heterodimers regulate striatal glutamatergic neurotransmission. However, few researches about kinetics have been reported. Here, we combined Iem-spFRET and E-FRET to investigate the kinetics of A1 R and A2A R interaction. Iem-spFRET obtains the energy transfer efficiency of the whole cell. E-FRET gets energy transfer efficiency with high spatial resolution, whereas, it was prone to biases because background was easily selected due to manual operation. To study the interaction with high spatio-temporal resolution, Iem-spFRET was used to correct the deviation of E-FRET. In this paper, A1 R and A2A R interaction was monitored, and the changes of FRET efficiency of the whole or/and partial cell membrane were described. The results showed that activation of A1 R or A2A R leads to rapid aggregation, inhibition of A1 R or A2A R leads to slow segregation, and the interaction is reversible. These results demonstrated that combination of Iem-spFRET and E-FRET could measure A1 R and A2A R interaction with high spatio-temporal resolution.

A microsphere nanoparticle based-serum albumin targeted adsorption coupled with surface-enhanced Raman scattering for breast cancer detection.

The serum albumin level is inseparable associated with survival in patients with breast cancer, and simultaneously serve as a good indicator of prognosis of cancer. Here, we proposed a novel extraction-isolation analysis method of albumin for breast cancer detection utilizing hydroxyapatite particles (HAp) to targeted adsorb albumin from serum relying on its specific adsorption capacity. An ideal protein-release reagent was used for isolating albumin from the surface of HAp, and meanwhile ensuring that the structure and property of albumin was not suffered damage. The surface-enhanced Raman scattering (SERS) signal of extracted albumin was obtained, and partial least squares (PLS) and linear discriminant analysis (LDA) analysis approach were employed to analyze SERS spectra data, with the aim to assess the capability of HAp method for identifying breast cancer, yielding an ideal diagnostic accuracy of 98.6%, demonstrating promising potential as a non-invasive and sensitive nanotechnology for breast cancer screening.

In vivo and in vitro study of co-expression of LMP1 and Cripto-1 in nasopharyngeal carcinoma / Estudo in vivo e in vitro da coexpressão de LMP1 e Cripto-1 em carcinoma nasofaríngeo

Abstract Introduction: Nasopharyngeal carcinoma, an epithelial-derived malignant tumor which because of its anatomical location and atypical early symptoms, when diagnosed invasion and metastasis often have occurred. This requires a better understanding of the development mechanism, identifying diagnostic markers, and developing new treatment strategies. Objective: To study the relationship of LMP1 and Cripto-1 in nasopharyngeal carcinoma. Methods: The expression of LMP1 and Cripto-1 in specimens obtained from nasopharyngeal carcinoma patients (n = 42) and nasopharyngitis patients (n = 22) were examined. The expression of LMP1 and Cripto-1 in LMP1-negative and LMP1-positive (CNE1-LMP1) cells were also examined. Results: The expression of LMP1 and Cripto-1 was significantly higher in nasopharyngeal carcinoma than in nasopharyngitis (p < 0.05). Their expression in nasopharyngeal carcinoma with metastasis were significantly higher than that without metastasis (p < 0.05), which was correlated with TNM staging (p < 0.05). High Cripto-1 expression and high proliferation rate were seen in CNE1-LMP1 cells. Conclusions: The expression of LMP1 and Cripto-1 in nasopharyngeal carcinoma is positively related. Their co-expression might contribute to the proliferation and metastasis of nasopharyngeal carcinoma.

Resumo Introdução: O carcinoma nasofaríngeo é um tumor maligno derivado do epitélio de localização anatômica recôndita e sintomas iniciais atípicos; quando diagnosticado, frequentemente invasão e metástases já ocorreram. Isso requer uma melhor compreensão do seu mecanismo de desenvolvimento, identificação dos marcadores diagnósticos e desenvolvimento de novas estratégias de tratamento. Objetivo: Estudar a relação de LMP1 e Cripto-1 no carcinoma nasofaríngeo. Método: A expressão de LMP1 e Cripto-1 em espécimes obtidos de pacientes com carcinoma de nasofaringe (n = 42) e pacientes com nasofaringite (n = 22) foi analisada. A expressão de LMP1 e Cripto-1 em células LMP1-negativas e LMP1-positivas (CNE1-LMP1) também foi analisada. Resultados: A expressão de LMP1 e Cripto-1 foi significantemente maior na presença de carcinoma nasofaríngeo do que na nasofaringite (p < 0,05). Sua expressão em carcinomas com metástase foi significantemente maior do que em casos sem metástase (p < 0,05), o que se correlacionou com o estadiamento TNM (p < 0,05). Uma alta expressão de Cripto-1 e alta taxa de proliferação foram observadas nas células CNE1-LMP1. Conclusões: A expressão de LMP1 e Cripto-1 é positivamente relacionada com carcinoma nasofaríngeo. Sua coexpressão pode ser atribuída à proliferação e metástase do tumor.

Toward image quality assessment in optical coherence tomography (OCT) of rat kidney.

BACKGROUND: Optical coherence tomography (OCT) is a useful tool for the evaluation of structure and function of the kidney, but the image quality can be effected by many factors. OBJECTIVE: The objective of this study was to assess the image quality of different OCT systems in OCT imaging of the living kidney. METHODS: One swept-source OCT (SSOCT) of 1300 nm, one spectral domain OCT (SDOCT) of 1300 nm and another of 900 nm were used. A FeO phantom was used to establish the point spread function (PSF). Rat kidneys were imaged for image quality assessment. Light penetration in the kidney and the optical attenuation coefficient were also evaluated. The quantification of uriniferous tubules was carried out via the threshold segmentation of 3D OCT images. RESULTS: The quality of kidney images was resolution dependent. SDOCT of 900 nm showed higher peak signal-to noise ratio and dynamic range. The spatial resolution in the light field could be derived from the PSF distribution along three mutually orthogonal axes. In conjunction with the PSF, the Lucy-Richardson algorithm could improve image quality but could not reveal more microstructural information. The penetration depth of 1300 nm was deeper than that of 900 nm. The attenuation coefficient of the kidney was 29 cm-1 at 1300 nm and 50 cm-1 at 900 nm (P < 0.001). More accurate measurement of uriniferous tubules was achieved with the SDOCT-900 due to its higher resolution. CONCLUSIONS: Both SSOCT and SDOCT systems could be useful for imaging uriniferous tubules in the superficial layers of the cortex. The OCT image quality was highly correlated with the spatial resolution of OCT system.

Attenuation of epigenetic regulator SMARCA4 and ERK-ETS signaling suppresses aging-related dopaminergic degeneration.

How complex interactions of genetic, environmental factors and aging jointly contribute to dopaminergic degeneration in Parkinson's disease (PD) is largely unclear. Here, we applied frequent gene co-expression analysis on human patient substantia nigra-specific microarray datasets to identify potential novel disease-related genes. In vivo Drosophila studies validated two of 32 candidate genes, a chromatin-remodeling factor SMARCA4 and a biliverdin reductase BLVRA. Inhibition of SMARCA4 was able to prevent aging-dependent dopaminergic degeneration not only caused by overexpression of BLVRA but also in four most common Drosophila PD models. Furthermore, down-regulation of SMARCA4 specifically in the dopaminergic neurons prevented shortening of life span caused by α-synuclein and LRRK2. Mechanistically, aberrant SMARCA4 and BLVRA converged on elevated ERK-ETS activity, attenuation of which by either genetic or pharmacological manipulation effectively suppressed dopaminergic degeneration in Drosophila in vivo. Down-regulation of SMARCA4 or drug inhibition of MEK/ERK also mitigated mitochondrial defects in PINK1 (a PD-associated gene)-deficient human cells. Our findings underscore the important role of epigenetic regulators and implicate a common signaling axis for therapeutic intervention in normal aging and a broad range of age-related disorders including PD.

Surface-enhanced Raman scattering analysis of serum albumin via adsorption-exfoliation on hydroxyapatite nanoparticles for noninvasive cancers screening.

Combining serum albumin via adsorption-exfoliation on hydroxyapatite particles (HAp) with surface-enhanced Raman scattering (SERS), we developed a novel quantitative analysis of albumin method from blood serum for cancers screening applications. The quantitatively analysis obtained by our HAp method had a good linear relationship from 1 to 10 g/dL, and the lower limit of detection was less than the albumin prognostic factor for disease (3.5 g/dL). Serum albumin was adsorbed and exfoliated by HAp from serum samples of liver cancer patients, breast cancer patients and healthy volunteers and mixed with silver colloids to perform SERS spectral analysis. Based on the PLS-SVM algorithm, the diagnostic accuracies of liver cancer patients and breast cancer patients were 100% and 96.68%, respectively. Moreover, this algorithm successfully predicted the unidentified subjects with a diagnostic accuracy of 93.75%. This exploratory work demonstrated that HAp-adsorbed-exfoliated serum proteins combined with SERS spectroscopy has great potential for cancer screening.

Optofluidic microbubble Fabry-Pérot cavity.

An optofluidic microbubble Fabry-Pérot (OMBFP) cavity was investigated. In contrast to plane-plane FP (PPFP) cavities, the optical mode confinement and stability in an OMBFP were significantly enhanced. The optical properties of the OMBFP cavity, including the quality (Q) factor, effective mode area, mode distribution as a function of the core refractive index, microbubble position, and mirror tilt angle, were investigated systematically using the finite element method. In optofluidic lasing experiments, a low lasing threshold of 1.25 µJ/mm2, which was one order magnitude lower than that of the PPFP, was achieved owing to improved modal lateral confinement. Since the microbubble acts as a micro-lens and microfluidic channel in the parallel FP cavity, mode selection and cell-dye laser were easily realized in the OMBFP cavity.

Examination of the relationship between viscoelastic properties and the invasion of ovarian cancer cells by atomic force microscopy.

The mechanical properties of cells could serve as an indicator for disease progression and early cancer diagnosis. This study utilized atomic force microscopy (AFM) to measure the viscoelastic properties of ovarian cancer cells and then examined the association with the invasion of ovarian cancer at the level of living single cells. Elasticity and viscosity of the ovarian cancer cells OVCAR-3 and HO-8910 are significantly lower than those of the human ovarian surface epithelial cell (HOSEpiC) control. Further examination found a dramatic increase of migration/invasion and an obvious decease of microfilament density in OVCAR-3 and HO-8910 cells. Also, there was a significant relationship between viscoelastic and biological properties among these cells. In addition, the elasticity was significantly increased in OVCAR-3 and HO-8910 cells after the treatment with the anticancer compound echinomycin (Ech), while no obvious change was found in HOSEpiC cells after Ech treatment. Interestingly, Ech seemed to have no effect on the viscosity of the cells. Ech significantly inhibited the migration/invasion and significantly increased the microfilament density in OVCAR-3 and HO-8910 cells, which was significantly related with the elasticity of the cells. An increase of elasticity and a decrease of invasion were found in OVCAR-3 and HO-8910 cells after Ech treatment. Together, this study clearly demonstrated the association of viscoelastic properties with the invasion of ovarian cancer cells and shed a light on the biomechanical changes for early diagnosis of tumor transformation and progression at single-cell level.

Coupled-mode induced transparency via Ohmic heating in a single polydimethylsiloxane-coated microbubble resonator.

We demonstrate an approach for the realization of coupled-mode induced transparency (CMIT) in a hybrid polydimethylsiloxane (PDMS)-coated silica microbubble resonator, with an Au microwire inserted in the hollow channel. Owing to the large negative thermo-optics coefficient of PDMS, different radial order modes with opposite thermal sensitivities can coexist in this hybrid microcavity. By applying a current through the Au microwire, which acts as a microheater, the generated Ohmic heating could thermally tune the resonance frequencies and the frequency detuning of the coupled mode to achieve controllable CMIT. This platform offers an efficient and convenient way to obtain controllable CMIT for applications, such as label-free biosensing and quantum information processing.

Tunable optofluidic liquid metal core microbubble resonator.

This study introduces design and coupling techniques, which bridge an opaque liquid metal, optical WGM mode, and mechanical mode into an opto-mechano-fluidic microbubble resonator (MBR) consisting of a dielectric silica shell and liquid metal core. Benefiting from the conductivity of the liquid metal, Ohmic heating was carried out for the MBR by applying current to the liquid metal to change the temperature of the MBR by more than 300 °C. The optical mode was thermally tuned (>3 nm) over a full free spectral range because the Ohmic heating changed the refractive index of the silica and dimeter of the MBR. The mechanical mode was thermally tuned with a relative tuning range of 9% because the Ohmic heating changed the velocity and density of the liquid metal.

Combining whispering gallery mode optofluidic microbubble resonator sensor with GR-5 DNAzyme for ultra-sensitive lead ion detection.

Lead ions are deleterious pollutants that often reach drinking water, and can cause significant harm to humans (particularly children). An ultra-sensitive lead ion detection method using a whispering gallery mode (WGM) optofluidic microbubble resonator and the classic GR-5 DNAzyme is proposed in this paper. With the auxiliary piranha and Ploy-l-lysine solution, GR-5 DNAzyme was successfully modified on the inner wall of a microbubble. The mode field distribution of the microbubble was analysed, and the optofluidic sensor with thin wall exhibited a maximum bulk refractive index sensitivity of 265.2 nm/RIU. Lead ions at concentrations ranging from 0.1 pM to 100 pM were tested using the proposed WGM optofluidic sensor. The noise was decreased to 2.43 fM using the self-referenced differential method. Thus, a limit of approximately 15 fM was obtained for the detection of lead ions using the WGM optofluidic biosensor. Eight competing metal ions were also used to evaluate the selectivity of the proposed sensor, with results indicating that it has high selectivity for lead ions. Finally, the sensor performance is verified using real samples.

In vivo and in vitro study of co-expression of LMP1 and Cripto-1 in nasopharyngeal carcinoma.

INTRODUCTION: Nasopharyngeal carcinoma, an epithelial-derived malignant tumor which because of its anatomical location and atypical early symptoms, when diagnosed invasion and metastasis often have occurred. This requires a better understanding of the development mechanism, identifying diagnostic markers, and developing new treatment strategies. OBJECTIVE: To study the relationship of LMP1 and Cripto-1 in nasopharyngeal carcinoma. METHODS: The expression of LMP1 and Cripto-1 in specimens obtained from nasopharyngeal carcinoma patients (n=42) and nasopharyngitis patients (n=22) were examined. The expression of LMP1 and Cripto-1 in LMP1-negative and LMP1-positive (CNE1-LMP1) cells were also examined. RESULTS: The expression of LMP1 and Cripto-1 was significantly higher in nasopharyngeal carcinoma than in nasopharyngitis (p<0.05). Their expression in nasopharyngeal carcinoma with metastasis were significantly higher than that without metastasis (p<0.05), which was correlated with TNM staging (p<0.05). High Cripto-1 expression and high proliferation rate were seen in CNE1-LMP1 cells. CONCLUSIONS: The expression of LMP1 and Cripto-1 in nasopharyngeal carcinoma is positively related. Their co-expression might contribute to the proliferation and metastasis of nasopharyngeal carcinoma.

Self-assembly of lipid rafts revealed by fluorescence correlation spectroscopy in living breast cancer cells.

Lipids and proteins in the plasma membrane are laterally heterogeneous and formalised as lipid rafts featuring unique biophysical properties. However, the self-assembly mechanism of lipid raft cannot be revealed even its physical properties and components were determined in specific physiological processes. In this study, two-photon generalised polarisation imaging and fluorescence correlation spectroscopy were used to study the fusion of lipid rafts through the membrane phase and the lateral diffusion of lipids in living breast cancer cells. A self-assembly model of lipid rafts associated with lipid diffusion and membrane phase was proposed to demonstrate the lipid sorting ability of lipid rafts in the plasma membrane. The results showed that the increased proportion of slow subdiffusion of GM1 -binding cholera toxin B-subunit (CT-B) was accompanied with an increased liquid-ordered domain during the ß-estradiol-induced fusion of lipid rafts. And slow subdiffusion of CT-B was vanished with the depletion of lipid rafts. Whereas the dialkylindocarbocyanine (DiIC18 ) diffusion was not specifically regulated by lipid rafts. This study will open up a new insight for uncovering the self-assembly of lipid rafts in specific pathophysiological processes.

Surface-enhanced Raman spectroscopy analysis of mast cell degranulation induced by low-intensity laser.

Mast cell (MC) degranulation is an important step in the healing process. In this study, silver-nanoparticles-based surface-enhanced Raman spectroscopy (SERS) was used to investigate the spectral characteristics of degranulation of MCs activated by low-intensity laser. The significant spectral changes, such as Raman peak intensities, suggested the concentration variation of some degranulated substances. The Raman intensity ratio of 799-554 cm-1 could be used as a potential internal indicator for the degranulation degree of MCs. Principal component analysis (PCA) was employed to reduce the high dimension of spectra into a few principal components (PCs) while retaining the most diagnostically significant information for sample differentiation. Using the diagnostically significant PC scores (P < 0.05), linear discriminate analysis (LDA) was applied to identify different cell degranulation groups with high sensitivity, specificity and accuracy. This exploratory work demonstrates that SERS technique combined with a PCA-LDA algorithm possesses great potential for developing a label-free, comprehensive, non-invasive and accurate method for measuring MC degranulation.

Precise closure of single blood vessels via multiphoton absorption-based photothermolysis.

We report a novel approach to selectively close single blood vessels within tissue using multiphoton absorption-based photothermolysis (multiphoton photothermolysis) without the need of exogenous agents. The treatment process is monitored by in vivo reflectance confocal microscopy in real time. Closure of single targeted vessels of varying sizes ranging from capillaries to venules was demonstrated. We also demonstrated that deeply situated blood vessels could be closed precisely while preserving adjacent overlying superficial blood vessels. In vivo confocal Raman spectroscopy of the treatment sites confirmed vessel closure as being mediated by local coagulative damage. Partial vessel occlusion could be achieved, and it is accompanied by increased intravascular blood cell speed. Multiphoton photothermolysis under real-time reflectance confocal imaging guidance provides a novel precision medicine approach for noninvasive, precise microsurgery treatment of vascular diseases on a per-vessel/per-lesion basis. The method could also be used for building ischemic stroke models for basic biology study.