A dynamic calcium-force relationship model for sag behavior in fast skeletal muscle.

In vitro studies using isolated or skinned muscle fibers suggest that the sigmoidal relationship between the intracellular calcium concentration and force production may depend upon muscle type and activity. The goal of this study was to investigate whether and how the calcium-force relationship changes during force production under physiological conditions of muscle excitation and length in fast skeletal muscles. A computational framework was developed to identify the dynamic variation in the calcium-force relationship during force generation over a full physiological range of stimulation frequencies and muscle lengths in cat gastrocnemius muscles. In contrast to the situation in slow muscles such as the soleus, the calcium concentration for the half-maximal force needed to drift rightward to reproduce the progressive force decline, or sag behavior, observed during unfused isometric contractions at the intermediate length under low-frequency stimulation (i.e., 20 Hz). The slope at the calcium concentration for the half-maximal force was required to drift upward for force enhancement during unfused isometric contractions at the intermediate length under high-frequency stimulation (i.e., 40 Hz). The slope variation in the calcium-force relationship played a crucial role in shaping sag behavior across different muscle lengths. The muscle model with dynamic variations in the calcium-force relationship also accounted for the length-force and velocity-force properties measured under full excitation. These results imply that the calcium sensitivity and cooperativity of force-inducing crossbridge formation between actin and myosin filaments may be operationally altered in accordance with the mode of neural excitation and muscle movement in intact fast muscles.

Optimizing Percutaneous Mitral Valvuloplasty for Rheumatic Mitral Stenosis　- Clinical Significance of Changes in Mitral Valve Area.

BACKGROUND: Percutaneous mitral valvuloplasty (PMV) is a standard treatment for severe rheumatic mitral stenosis (RMS). However, the prognostic significance of the change in mitral valve area (∆MVA) during PMV is not fully understood.Methods and Results: This study analyzed data from the Multicenter mitrAl STEnosis with Rheumatic etiology (MASTER) registry, which included 3,140 patients with severe RMS. We focused on patients with severe RMS undergoing their first PMV. Changes in echocardiographic parameters, including MVA quantified before and after PMV, and composite outcomes, including mitral valve reintervention, heart failure admission, stroke, and all-cause death, were evaluated. An optimal result was defined as a postprocedural MVA ≥1.5 cm2without mitral regurgitation greater than Grade II. Of the 308 patients included in the study, those with optimal results and ∆MVA >0.5 cm² had a better prognosis (log-rank P<0.001). Patients who achieved optimal results but with ∆MVA ≤0.5 cm² had a greater risk of composite outcomes than those with optimal outcomes and ∆MVA >0.5 cm² (nested Cox regression analysis, hazard ratio 2.27; 95% confidence interval 1.09-4.73; P=0.028). CONCLUSIONS: Achieving an increase in ∆MVA of >0.5 cm2was found to be correlated with improved outcomes. This suggests that, in addition to achieving traditional optimal results, targeting an increase in ∆MVA of >0.5 cm2could be a beneficial objective in PMV treatment for RMS.

Shifts in Clinical Characteristics, Treatment, and Outcome for Rheumatic Mitral Stenosis: Insights From a 20-Year Multicentre Registry Study in Korea.

BACKGROUND: The rapid economic development of South Korea provides a unique model to study changes in the clinical characteristics, treatment approaches, and clinical outcomes of patients with rheumatic mitral stenosis (MS) relative to socioeconomic growth. METHODS: From the Multicenter mitrAl STEnosis with Rheumatic etiology (MASTER) registry, 2,337 patients diagnosed with moderate or severe rheumatic MS between January 2001 and December 2020 were analyzed. Patients were grouped into consecutive 5-year intervals based on their year of diagnosis. Clinical characteristics, echocardiographic data, and clinical outcomes were assessed. RESULTS: Over 20 years, the severity of mitral stenosis increased from 79.1% to 90.2%; similarly, the average age at diagnosis increased from 54.3 to 63.0 years (all P < 0.001). Comorbidities such as hypertension and atrial fibrillation increased (6.3% to 29.5% and 41.4% to 46.9%, respectively; all P for trend < 0.05). The rate of mitral intervention within five years after diagnosis increased from 31.2% to 47.4% (P for trend < 0.001). However, clinical outcomes of rheumatic mitral stenosis deteriorated over time in the composite outcomes (log-rank test, P < 0.001). Conversely, the incidence of stroke remained stable (60.6-73.7%; P < 0.001), which might be attributed to the increased use of anticoagulation therapy. CONCLUSION: This study observed an increase in patient age, comorbidities, and valve disease severity as the country transitioned from a developing to developed status. Despite a rise in mitral valve interventions, clinical outcomes deteriorated over 20 years, highlighting the need for modified treatment approaches to improve patient outcomes.

High-Resolution Tactile-Sensation Diagnostic Imaging System for Thyroid Cancer.

In this study, we propose the direct diagnosis of thyroid cancer using a small probe. The probe can easily check the abnormalities of existing thyroid tissue without relying on experts, which reduces the cost of examining thyroid tissue and enables the initial self-examination of thyroid cancer with high accuracy. A multi-layer silicon-structured probe module is used to photograph light scattered by elastic changes in thyroid tissue under pressure to obtain a tactile image of the thyroid gland. In the thyroid tissue under pressure, light scatters to the outside depending on the presence of malignant and positive properties. A simple and easy-to-use tactile-sensation imaging system is developed by documenting the characteristics of the organization of tissues by using non-invasive technology for analyzing tactile images and judging the properties of abnormal tissues.

Decreased hemoglobin levels, cerebral small-vessel disease, and cortical atrophy: among cognitively normal elderly women and men.

BACKGROUND: Decreased hemoglobin levels increase the risk of developing dementia among the elderly. However, the underlying mechanisms that link decreased hemoglobin levels to incident dementia still remain unclear, possibly due to the fact that few studies have reported on the relationship between low hemoglobin levels and neuroimaging markers. We, therefore, investigated the relationships between decreased hemoglobin levels, cerebral small-vessel disease (CSVD), and cortical atrophy in cognitively healthy women and men. METHODS: Cognitively normal women (n = 1,022) and men (n = 1,018) who underwent medical check-ups and magnetic resonance imaging (MRI) were enrolled at a health promotion center. We measured hemoglobin levels, white matter hyperintensities (WMH) scales, lacunes, and microbleeds. Cortical thickness was automatically measured using surface based methods. Multivariate regression analyses were performed after controlling for possible confounders. RESULTS: Decreased hemoglobin levels were not associated with the presence of WMH, lacunes, or microbleeds in women and men. Among women, decreased hemoglobin levels were associated with decreased cortical thickness in the frontal (Estimates, 95% confidence interval, -0.007, (-0.013, -0.001)), temporal (-0.010, (-0.018, -0.002)), parietal (-0.009, (-0.015, -0.003)), and occipital regions (-0.011, (-0.019, -0.003)). Among men, however, no associations were observed between hemoglobin levels and cortical thickness. CONCLUSION: Our findings suggested that decreased hemoglobin levels affected cortical atrophy, but not increased CSVD, among women, although the association is modest. Given the paucity of modifiable risk factors for age-related cognitive decline, our results have important public health implications.

Association between body mass index and cortical thickness: among elderly cognitively normal men and women.

BACKGROUND: There is increasing evidence of a relationship between underweight or obesity and dementia risk. Several studies have investigated the relationship between body weight and brain atrophy, a pathological change preceding dementia, but their results are inconsistent. Therefore, we aimed to evaluate the relationship between body mass index (BMI) and cortical atrophy among cognitively normal participants. METHODS: We recruited cognitively normal participants (n = 1,111) who underwent medical checkups and detailed neurologic screening, including magnetic resonance imaging (MRI) in the health screening visits between September 2008 and December 2011. The main outcome was cortical thickness measured using MRI. The number of subjects with five BMI groups in men/women was 9/9, 148/258, 185/128, 149/111, and 64/50 in underweight, normal, overweight, mild obesity, and moderate to severe obesity, respectively. Linear and non-linear relationships between BMI and cortical thickness were examined using multiple linear regression analysis and generalized additive models after adjustment for potential confounders. RESULTS: Among men, underweight participants showed significant cortical thinning in the frontal and temporal regions compared to normal weight participants, while overweight and mildly obese participants had greater cortical thicknesses in the frontal region and the frontal, temporal, and occipital regions, respectively. However, cortical thickness in each brain region was not significantly different in normal weight and moderate to severe obesity groups. Among women, the association between BMI and cortical thickness was not statistically significant. CONCLUSIONS: Our findings suggested that underweight might be an important risk factor for pathological changes in the brain, while overweight or mild obesity may be inversely associated with cortical atrophy in cognitively normal elderly males.

Termination of the ligamentum venosum and the topographic relationship between the left portal vein, left hepatic artery, and ligamentum venosum in the fissures for the ligamentum teres and ligamentum venosum.

PURPOSE: The aim of this study was to clarify the location of termination of the ligamentum venosum (LV) and topographic relationships between the left portal vein (LPV), left hepatic artery (LHA), and LV in the fissures for ligamentum teres and LV. METHODS: Anatomical examination was performed on 50 livers from embalmed Korean adult cadavers. RESULTS: The LV originated from a point between the transverse and umbilical portions of LPV (Tr-Um point) in all specimens, and terminated in the common trunk of the middle hepatic vein and left hepatic vein (LHV) (61%) or in the LHV (39%). In the fissures for ligamentum teres and LV, the courses of the LHA relative to the LV and the LPV varied, such that four main patterns could be discerned. In type I (48%), the LHA crossed the inferior surface of Tr-Um point and passed just left of the origin of LV. In type II (20%), the LHA crossed the inferior surface of umbilical portion of LPV. In type III (10%), the LHA crossed the inferior surface of Tr-Um point to pass just left of origin of LV, with one branch of the right hepatic artery crossing the inferior surface of umbilical portion of LPV. In type IV (12%), no arterial branches ran on the inferior surfaces of transverse and umbilical portions of LPV. CONCLUSION: These details may play a role as external landmarks for facilitating preestimation of intrahepatic structures during various surgical procedures involving the liver.

New insights into the origin of the lumbrical muscles of the foot: tendinous slip of the flexor hallucis longus muscle.

PURPOSE: The aim of this study was to clarify the origins of the lumbricals of the foot toward a better understanding of its precise action in the gait. METHODS: The lumbricals in the foot were investigated in 66 specimens of embalmed Korean adult cadavers. RESULTS: The first lumbrical arose as two muscle bellies from both the tendon of the FDL and the tendinous slip of the FHL in 83.3 %, and as one muscle belly from the tendon of the FDL or the tendinous slip of the FHL in 16.7 %. These two muscle bellies subsequently merged to form the muscle belly of the first lumbrical. The second lumbrical arose from the tendinous slips of the FHL for the second and third toes as well as the tendon of the FDL in all specimens. The third lumbrical arose from the tendinous slips of the FHL for the third and fourth toes in 69.7 %, and the fourth lumbrical arose from the tendinous slip of the FHL for the fourth toe in 18.2 %. Some deep muscle fibers of the fourth lumbrical arose from the tendinous slip of the FHL for the second toe in 4.5 %, for the third toe in 28.8 %, and for the fourth toe in 15.2 %. CONCLUSIONS: The results of this study constitute new anatomical knowledge regarding the origin of the lumbricals, and provide insight into their specific role in production of gait. These findings will be useful for various types of surgery, biomechanics research, and rehabilitation programs.

Blending of the lateral deep slip of the platysma muscle into the buccinator muscle.

PURPOSE: The aim of this study was to clarify the prevalence of blending of the lateral deep slip of the platysma into the buccinator, and the morphologic and spatial relationships thereof. METHODS: The lateral deep slip of the platysma was investigated in 42 specimens of embalmed adult Korean cadavers. RESULTS: The lateral deep slip of the platysma was observed in 17 of the 42 specimens (40.5 %). When present, it divided from the deep portion (16 specimens, 38.1 %) or the lateral portion (1 specimen, 2.4 %) of the platysma pars modiolaris. The lateral deep slip passed deep to the facial artery, where it blended into the inferior part of the buccinator. The blending site was located inferolateral to the modiolus. After blending into the buccinator, the fibers of the lateral deep slip coursed longitudinally with the inferior fibers of the buccinator. CONCLUSION: The present description of the topography of the lateral deep slip of the platysma and its relationship with surrounding structures could improve the understanding of the detailed movements of the lower face and the related EMG activities, with the data also being useful when performing facial surgery.

An anatomic study of the medial fibers of depressor anguli oris muscle passing deep to the depressor labii inferioris muscle.

The trajectory of the medial fibers of depressor anguli oris muscle that pass deep to the depressor labii inferioris muscle (dDAO) and the anatomic relationship have not yet been examined. The aim of this study was to clarify the arrangement of the dDAO and its relationships with the surrounding structures. The dDAOs were investigated in 40 hemifaces from Korean cadavers. The dDAOs were observed in 18 of the 40 specimens (45%). The anatomic relationships of the dDAO were classified into the following 4 categories according to their connection with the incisivus labii inferioris muscle (ILI): (1) connected to the ILI and attached to the inferior border of the mandible lateral to the mentalis muscle (14 specimens, 35%), (2) connected to the ILI and intermingled with the lateral lower fibers of the mentalis muscle (2 specimens, 5%), (3) connected to the ILI but neither attached to the mandible nor intermingled with the mentalis muscle (1 specimen, 2.5%), and (4) not connected to the ILI but intermingled with the mentalis muscle (1 specimen, 2.5%). Therefore, in 17 specimens, the dDAOs were connected to the ILI (42.5%), in 14 specimens, they were attached to the inferior border of the mandible (35%), and, in 3 specimens, they were intermingled with the mentalis muscle (7.5%). The new anatomic data regarding the dDAO will contribute toward the understanding of movements of the mouth and lower face, which will be helpful when designing effective botulinum toxin type A therapies and performing various types of facial surgery.

Long-lasting forms of plasticity through patterned ultrasound-induced brainwave entrainment.

Achieving long-lasting neuronal modulation with low-intensity, low-frequency ultrasound is challenging. Here, we devised theta burst ultrasound stimulation (TBUS) with gamma bursts for brain entrainment and modulation of neuronal plasticity in the mouse motor cortex. We demonstrate that two types of TBUS, intermittent and continuous TBUS, induce bidirectional long-term potentiation or depression-like plasticity, respectively, as evidenced by changes in motor-evoked potentials. These effects depended on molecular pathways associated with long-term plasticity, including N-methyl-d-aspartate receptor and brain-derived neurotrophic factor/tropomyosin receptor kinase B activation, as well as de novo protein synthesis. Notably, bestrophin-1 and transient receptor potential ankyrin 1 play important roles in these enduring effects. Moreover, pretraining TBUS enhances the acquisition of previously unidentified motor skills. Our study unveils a promising protocol for ultrasound neuromodulation, enabling noninvasive and sustained modulation of brain function.

Outcomes of Severe Mitral Stenosis With the Revised Severity Criteria: Mitral Valve Replacement vs Percutaneous Mitral Valvuloplasty.

BACKGROUND: This study aimed to compare the outcomes, according to percutaneous mitral valvuloplasty (PMV) vs mitral valve replacement (MVR), of severe mitral stenosis (MS) with the updated criteria (MVA ≤ 1.5 cm2). METHODS: From the Multicenter Mitral Stenosis With Rheumatic Etiology (MASTER) registry of 3140 patients, we included patients with severe MS who underwent PMV or MVR between January 2000 and December 2021 except for previous valvular surgery/intervention, at least moderate other valvular dysfunction, and thrombus at the left atrium/appendage. Moderately severe MS (MS-MS) and very severe MS (VS-MS) were defined as 1.0 cm2 < MVA ≤ 1.5 cm2 and MVA ≤ 1.0 cm2, respectively. Primary outcomes were a composite of cardiovascular (CV) death and heart failure (HF) hospitalization. Secondary outcomes were a composite of primary outcomes and redo intervention. RESULTS: Among 442 patients (mean 56.5 ±11.9 years, women 77.1%), the MVR group (n = 260) was older, had more comorbidities, higher echoscore, larger left chambers, and higher right ventricular systolic pressure than the PMV group (n = 182). During a mean follow-up of 6.9 ± 5.2 years with inverse probability-weighted matching, primary outcomes did not differ, but the MVR group experienced fewer secondary outcomes (P = 0.010). In subgroup analysis of patients with MS-MS and VS-MS, primary outcomes did not differ. However, the MVR group in patients with VS-MS showed better secondary outcomes (P = 0.012). CONCLUSIONS: PMV or MVR did not influence CV mortality or HF hospitalization in both MS-MS and VS-MS. However, because of increased early redo intervention in the PMV group in VS-MS, MVR would be the preferable option without clear evidence of suitable morphology for PMV.

Morphology of the mentalis muscle and its relationship with the orbicularis oris and incisivus labii inferioris muscles.

The mentalis muscle (MT) is the only elevator of the lower lip and the chin, and it provides the major vertical support for the lower lip. However, there are few reports on the relationship between the MT and its surrounding muscles. The aim of this study was to clarify the morphology of the MT, especially in relation to the orbicularis oris muscle and the incisivus labii inferioris muscle (ILI), thereby providing data to understand the function of the MT in relation to the surrounding muscles. The MT was examined in 40 specimens of embalmed adult Korean cadavers. The medial fibers of both MTs descended anteromedially and crossed together, forming a dome-shaped chin prominence in all specimens. The lateral fibers of the MT descended inferomedially in 38 specimens (95%) and inferolaterally in 2 specimens (5%). The upper fibers of the MT were short and ran horizontally, whereas the lower fibers were long and descended inferomedially or vertically. The upper fibers of the MT were intermingled with the inferior margin of the orbicularis oris muscle in all specimens. The originating muscle fibers of the ILI were intermingled with the upper lateral MT in all specimens. Some of the ILI fibers extended inferomedially to the middle or lower portion of the MT in 22 specimens (55%). The results of this study constitute new anatomical knowledge regarding the MT and will be useful to surgeons performing various surgical procedures of the chin area.

Decoding force production of skeletal muscle from the female brain using functional near-infrared spectroscopy.

OBJECTIVE: Noninvasive neural decoding enables predicting motor output from neural activities without physically damaging the human body. A recent study demonstrated the applicability of functional near-infrared spectroscopy (fNIRS) to decode muscle force production from hemodynamic signals measured in the male brain. However, given the sex differences in cerebral blood flow and muscle physiology, whether the fNIRS approach can also be applied to the female brain remains elusive. Therefore, this study aimed to evaluate whether fNIRS can be used to identify the optimal cortical region and hemodynamic predictor to decode muscle force output in females. RESULTS: Statistical group analysis for eight healthy female adults showed that the cortical region for wrist control was topologically dorsal to that for finger control over the primary sensorimotor cortex. This cortical area was maximally activated while the wrist flexor muscles were contracted to hold a load on the subject's palm, as was the case for males. However, the dynamics of oxyhemoglobin concentration measured from the most activated cortical area differed between females and males. The signal intensity during 100% maximal voluntary contraction and the signal increase rate at 50% maximal voluntary contraction was lower and faster in females. Eight predictors were used to characterize hemodynamic signals' amplitude and temporal variation in the female cortex. Unlike the case for males, only the trajectory predictors for the amplitude of oxyhemoglobin concentration change were strongly correlated with the strengths of force produced by the wrist flexor muscles, showing a linear relationship. These results suggest gender-specific hemodynamics must be considered for decoding low-level motor control with fNIRS in females.

Asymmetric electrotonic coupling between the soma and dendrites alters the bistable firing behaviour of reduced models.

The goal of the study was to investigate the influence of asymmetric coupling, between the soma and dendrites, on the nonlinear dynamic behaviour of a two-compartment model. We used a recently published method for generating reduced two-compartment models that retain the asymmetric coupling of anatomically reconstructed motor neurons. The passive input-output relationship of the asymmetrically coupled model was analytically compared to the symmetrically coupled case. Predictions based on the analytic comparison were tested using numerical simulations. The simulations evaluated the nonlinear dynamics of the models as a function of coupling parameters. Analytical results showed that the input resistance at the dendrite of the asymmetric model was directly related to the degree of coupling asymmetry. In contrast, a comparable symmetric model had identical input resistances at both the soma and dendrite regardless of coupling strength. These findings lead to predictions that variations in dendritic excitability, subsequent to changes in input resistance, might change the current threshold and onset timing of the plateau potential generated in the dendrite. Since the plateau potential underlies bistable firing, these results further predicted that asymmetric coupling might alter nonlinear (i.e. bistable) firing patterns. The numerical simulations supported analytical predictions, showing that the fully bistable firing pattern of the asymmetric model depended on the degree of coupling asymmetry and its correlated dendritic excitability. The physiological property of asymmetric coupling plays an important role in generating and stabilizing the bistability of motor neurons by interacting with the excitability of dendritic branches.

Destination of the C4 component of the prefixed brachial plexus.

This study was carried out to elucidate the destination of the C4 ventral ramus that forms the prefixed brachial plexus. A total of 43 sides of Korean adults (male, 22; female, 18; unknown: 3 and right, 23; left, 20) were investigated. The various C4 destinations and their frequencies were as follows: suprascapular nerve, 97.7% (42/43); musculocutaneous nerve, 79.1% (34/43); the axillary nerve, 48.8% (21/43); and the nerve to subclavius, 16.2% (7/43). The mean quantity of nerve fibers destined to the suprascapular, musculocutaneous, and axillary nerves and the nerve to subclavius was 441.8 ± 316.5 (Mean ± S.D.), 298.2 ± 209.8, 245.8 ± 239.0, and 66.4 ± 17.2, respectively. The mean nerve fibers number of C4 contributing to the brachial plexus was 983.6 ± 618.0 (range: 100-1729). The main component that contributes to the terminal nerves (suprascapular, musculocutaneous, axillary nerves, and nerve to subclavius) is C5. These results may be useful to surgeons and clinicians dealing with terminal nerves of the brachial plexus, especially the suprascapular, musculocutaneous, and axillary nerves.

Cerebral hemodynamics predicts the cortical area and coding scheme in the human brain for force generation by wrist muscles.

The goal of this study is to identify the cortical area maximally active over the primary sensorimotor cortex (SM1) and characterize the cortical encoding for force production by wrist muscles in the human brain. The technique of functional near-infrared spectroscopy (fNIRS) was used to continuously monitor the changes in hemoglobin concentrations from the left hemisphere during isometric contractions of wrist flexion muscles over a broad range of load forces (0 ∼ 8 kgf) on the right hand. As previously shown in primate studies, this action produced hemodynamic activity predominantly in the wrist area localized dorsally to the finger region over SM1 and the hemodynamic response was systematically related to the level of load intensity. The coding scheme for force production in terms of hemodynamic signals was characterized defining eight trajectory parameters (four for amplitude coding and four for temporal coding) and analyzed for the area maximally activated over SM1. The trajectory parameter representing the oxygenated hemoglobin concentration change at the end of motor task (amplitude coding) and the timing of maximum change in oxygenated hemoglobin concentration (temporal coding) was most strongly correlated with the load variation in a superliner manner. All these results indicate the applicability of fNIRS to monitor and decode cortical activity that is correlated with low-level motor control such as isometric muscle contractions. This study may provide not only insights into cortical neural control of muscle force but also predictors of muscle force in clinical diagnostics and neural interfaces for the human brain.

Effective Stimulation Type and Waveform for Force Control of the Motor Unit System: Implications for Intraspinal Microstimulation.

The input-output properties of spinal motoneurons and muscle fibers comprising motor units are highly non-linear. The goal of this study was to investigate the stimulation type (continuous versus discrete) and waveform (linear versus non-linear) controlling force production at the motor unit level under intraspinal microstimulation. We constructed a physiological model of the motor unit with computer software enabling virtual experiments on single motor units under a wide range of input conditions, including intracellular and synaptic stimulation of the motoneuron and variation in the muscle length under neuromodulatory inputs originating from the brainstem. Continuous current intensity and impulse current frequency waveforms were inversely estimated such that the motor unit could linearly develop and relax the muscle force within a broad range of contraction speeds and levels during isometric contraction at various muscle lengths. Under both continuous and discrete stimulation, the stimulation waveform non-linearity increased with increasing speed and level of force production and with decreasing muscle length. Only discrete stimulation could control force relaxation at all muscle lengths. In contrast, continuous stimulation could not control force relaxation at high contraction levels in shorter-than-optimal muscles due to persistent inward current saturation on the motoneuron dendrites. These results indicate that non-linear adjustment of the stimulation waveform is more effective in regard to varying the force profile and muscle length and that the discrete stimulation protocol is a more robust approach for designing stimulation patterns aimed at neural interfaces for precise movement control under pathological conditions.

Coculture with BJ fibroblast cells inhibits the adipogenesis and lipogenesis in 3T3-L1 cells.

Mouse or human fibroblasts are commonly used as feeder cells to prevent differentiation in stem or primary cell culture. In the present study, we addressed whether fibroblasts can affect the differentiation of adipocytes. We found that the differentiation of 3T3-L1 preadipocytes was strongly suppressed when the cells were cocultured with human fibroblast (BJ) cells. BrdU incorporation analysis indicated that mitotic clonal expansion, an early event required for 3T3-L1 cell adipogenesis, was not affected by BJ cells. The 3T3-L1 cell expression levels of peroxisome proliferator-activated receptor gamma2, CCAAT/enhancer-binding protein alpha (C/EBPalpha), sterol regulatory element binding protein-1c, and Krüppel-like factor 15, but not those of C/EBPbeta or C/EBPdelta, were decreased by coculture with BJ cells. When mature 3T3-L1 adipocytes were cocultured with BJ cells, their lipid contents were significantly reduced, with decreased fatty acid synthase expression and increased phosphorylated form of acetyl-CoA carboxylase 1. Our data indicate that coculture with BJ fibroblast cells inhibits the adipogenesis of 3T3-L1 preadipocytes and decreases the lipogenesis of mature 3T3-L1 adipocytes.

Screening of Kit inhibitors: suppression of Kit signaling and melanogenesis by emodin.

In search of novel antipigmentation agents, a set of 3,840 compounds with natural-like synthetic or natural origin were screened against Kit (stem cell factor receptor). Emodin from the seed of Cassia tora and baicalin from Scutellariae radix showed potent inhibitory effects (IC(50) = 4.9 and 9.0 microM, respectively) on the phosphorylation of Kit. Emodin also blocked other receptor tyrosine kinase activities, such as epithelial growth factor receptor (EGFR), vascular endothelial growth factor receptor 2 (VEGFR-2), fibroblast growth factor receptor 1 (FGFR-1), platelet-derived growth factor receptor b (PDGFR-b). In contrast to emodin, aloe-emodin did not inhibit Kit activity at all. Emodin also blocked the cellular kinase activities of Kit and its down-stream p44/42 mitogen activated protein kinase (MAPK) in MO7e cells and human primary melanocytes. Emodin strongly suppressed the melanin synthesis triggered by stem cell factor (SCF) treatment. Also, emodin showed almost no toxicity up to 10 microM on cultured melanocytes as reported previously by other researchers. The results indicate that emodin is a good candidate for the development of antipigmentation agents since it can radically block the differentiation and proliferation of pigment cells by reducing Kit signaling.