Early functional factors for predicting outcome of independence in daily living after stroke: a decision tree analysis.

OBJECTIVE: This study aimed to investigate the predictive functional factors influencing the acquisition of basic activities of daily living performance abilities during the early stages of stroke rehabilitation using classification and regression analysis trees. METHODS: The clinical data of 289 stroke patients who underwent rehabilitation during hospitalization (164 males; mean age: 62.2 ± 13.9 years) were retrospectively collected and analysed. The follow-up period between admission and discharge was approximately 6 weeks. Medical records, including demographic characteristics and various functional assessments with item scores, were extracted. The modified Barthel Index on discharge served as the target outcome for analysis. A "good outcome" was defined as a modified Barthel Index score ≥ 75 on discharge, while a modified Barthel Index score < 75 was classified as a "poor outcome." RESULTS: Two classification and regression analysis tree models were developed. The first model, predicting activities of daily living outcomes based on early motor functions, achieved an accuracy of 92.4%. Among patients with a "good outcome", 70.9% exhibited (i) ≥ 4 points in the "sitting-to-standing" category in the motor assessment scale and (ii) 32 points on the Berg Balance Scale score. The second model, predicting activities of daily living outcome based on early cognitive functions, achieved an accuracy of 82.7%. Within the "poor outcome" group, 52.2% had (i) ≤ 21 points in the "visuomotor organization" category of Lowenstein Occupational Therapy Cognitive Assessment, (ii) ≤ 1 point in the "time orientation" category of the Mini Mental State Examination. CONCLUSION: The ability to perform "sitting-to-standing" and visuomotor organization functions at the beginning of rehabilitation emerged as the most significant predictors for achieving successful basic activities of daily living on discharge after stroke.

Comparison of biomechanical parameters in lower limb joints of stroke patients according to conventional evaluation scores during level walking.

Objective: Patients with chronic stroke capable of independent gait were classified into functional ambulation category (FAC) 4 or 5, and the kinetic and kinematic data on their lower limb joints on the affected and unaffected sides were compared with that of healthy individuals. Finally, the qualitative changes in the gait of patients with stroke were investigated based on the differences in FAC scores. Methods: Twelve healthy participants and 19 patients with stroke capable of independent gait were included. The three-dimensional (3D) motion analysis and conventional assessment were conducted for all patients with stroke. Results: The FAC 5 group exhibited a larger range of motion (ROM) than the FAC 4 group in knee and hip joints on the affected side and only in the hip on the unaffected side. In the FAC 5 group, ROM differences in the healthy group on either the affected or unaffected side were absent. The peak of the hip flexion moment on the affected side in both the FAC 4 and 5 groups was smaller than that in the healthy group and in the FAC 4 group on the unaffected side. The absorption power minimum on the affected side was smaller only in the FAC 4 group than that in the healthy group and was larger in the FAC 5 group than that in the FAC 4 group. On the unaffected side, the absorption power minimum was smaller only in the FAC 4 group than that in the healthy group. Conclusion: Functional differences in gait were found in patients classified based on conventional evaluation capable of independent gait after post-stroke rehabilitation. Patients may not exhibit complete recovery in the kinetic indices even if they are judged to be normal in the conventional evaluation, and the kinematic gait indices indicate recovery. Evaluating kinetic indices in addition to kinematic indices is necessary, and joint power may be an especially useful index.

Alterations in learning-related cortical activation and functional connectivity by high-definition transcranial direct current stimulation after stroke: an fNIRS study.

Introduction: Motor learning is a key component of stroke neurorehabilitation. High-definition transcranial direct current stimulation (HD-tDCS) was recently developed as a tDCS technique that increases the accuracy of current delivery to the brain using arrays of small electrodes. The purpose of this study was to investigate whether HD-tDCS alters learning-related cortical activation and functional connectivity in stroke patients using functional near-infrared spectroscopy (fNIRS). Methods: Using a sham-controlled crossover study design, 16 chronic stroke patients were randomly assigned to one of two intervention conditions. Both groups performed the sequential finger tapping task (SFTT) on five consecutive days, either with (a) real HD-tDCS or (b) with sham HD-tDCS. HD-tDCS (1 mA for 20 min, 4 × 1) was administered to C3 or C4 (according to lesion side). fNIRS signals were measured during the SFTT with the affected hand before (baseline) and after each intervention using fNIRS measurement system. Cortical activation and functional connectivity of NIRS signals were analyzed using a statistical parametric mapping open-source software package (NIRS-SPM), OptoNet II®. Results: In the real HD-tDCS condition, oxyHb concentration increased significantly in the ipsilesional primary motor cortex (M1). Connectivity between the ipsilesional M1 and the premotor cortex (PM) was noticeably strengthened after real HD-tDCS compared with baseline. Motor performance also significantly improved, as shown in response time during the SFTT. In the sham HD-tDCS condition, functional connectivity between contralesional M1 and sensory cortex was enhanced compared with baseline. There was tendency toward improvement in SFTT response time, but without significance. Discussion: The results of this study indicated that HD-tDCS could modulate learning-related cortical activity and functional connectivity within motor networks to enhance motor learning performance. HD-tDCS can be used as an additional tool for enhancing motor learning during hand rehabilitation for chronic stroke patients.

Task-Related Hemodynamic Changes Induced by High-Definition Transcranial Direct Current Stimulation in Chronic Stroke Patients: An Uncontrolled Pilot fNIRS Study.

High-definition transcranial direct current stimulation (HD-tDCS) has recently been proposed as a tDCS approach that can be used on a specific cortical region without causing undesirable stimulation effects. In this uncontrolled pilot study, the cortical hemodynamic changes caused by HD-tDCS applied over the ipsilesional motor cortical area were investigated in 26 stroke patients. HD-tDCS using one anodal and four cathodal electrodes at 1 mA was administered for 20 min to C3 or C4 in four daily sessions. Cortical activation was measured as changes in oxyhemoglobin (oxyHb) concentration, as found using a functional near-infrared spectroscopy (fNIRS) system during the finger tapping task (FTT) with the affected hand before and after HD-tDCS. Motor-evoked potential and upper extremity functions were also measured before (T0) and after the intervention (T1). A group statistical parametric mapping analysis showed that the oxyHb concentration increased during the FTT in both the affected and unaffected hemispheres before HD-tDCS. After HD-tDCS, the oxyHb concentration increased only in the affected hemisphere. In a time series analysis, the mean and integral oxyHb concentration during the FTT showed a noticeable decrease in the channel closest to the hand motor hotspot (hMHS) in the affected hemisphere after HD-tDCS compared with before HD-tDCS, in accordance with an improvement in the function of the affected upper extremity. These results suggest that HD-tDCS might be helpful to rebalance interhemispheric cortical activity and to reduce the hemodynamic burden on the affected hemisphere during hand motor tasks. Noticeable changes in the area adjacent to the affected hMHS may imply that personalized HD-tDCS electrode placement is needed to match each patient's individual hMHS location.

Whole Brain Hemodynamic Response Based on Synchrony Analysis of Brain Signals for Effective Application of HD-tDCS in Stroke Patients: An fNIRS Study.

In this study, the effective application of high-definition transcranial direct current stimulation (HD-tDCS) based on the whole brain hemodynamic response in stroke patients was investigated using functional near-infrared spectroscopy (fNIRS). The intrahemispheric and interhemispheric synchronization and cortical activity based on the time during 1 mA HD-tDCS were examined in 26 chronic cerebrovascular disease patients. At the beginning of HD-tDCS, the synchronization and brain activity in the whole brain increased rapidly and decreased after 5 min. In the middle of tDCS, the synchronization began to increase again, and strong synchronic connections were formed around the desired stimulation area. After tDCS, strong cortical activation was observed in the stimulation area, indicating that the baseline of the oxyhemoglobin (HbO) signal increased in the desired stimulation area. Therefore, the results of this study indicate that HD-tDCS can be applied efficiently to enhance the effect of tDCS. This stimulation method with tDCS can be explored clinically for more neurorehabilitation of patients with degenerative brain diseases.

Ferroelastic-Ferroelectric Multiferroicity in van der Waals Rhenium Dichalcogenides.

2D multiferroics with combined ferroic orders have gained attention owing to their novel functionality and underlying science. Intrinsic ferroelastic-ferroelectric multiferroicity in single-crystalline van der Waals rhenium dichalcogenides, whose symmetries are broken by the Peierls distortion and layer-stacking order, is demonstrated. Ferroelastic switching of the domain orientation and accompanying anisotropic properties is achieved with 1% uniaxial strain using the polymer encapsulation method. Based on the electron localization function and bond dissociation energy of the Re-Re bonds, the change in bond configuration during the evolution of the domain wall and the preferred switching between the two specific orientation states are explained. Furthermore, the ferroelastic switching of ferroelectric polarization is confirmed using the photovoltaic effect. The study provides insights into the reversible bond-switching process and potential applications based on 2D multiferroicity.

Multimodal Imaging Biomarker-Based Model Using Stratification Strategies for Predicting Upper Extremity Motor Recovery in Severe Stroke Patients.

Background. Various prognostic biomarkers for upper extremity (UE) motor recovery after stroke have been reported. However, most have relatively low predictive accuracy in severe stroke patients.Objective. This study suggests an imaging biomarker-based model for effectively predicting UE recovery in severe stroke patients.Methods. Of 104 ischemic stroke patients screened, 42 with severe motor impairment were included. All patients underwent structural, diffusion, and functional magnetic resonance imaging at 2 weeks and underwent motor function assessments at 2 weeks and 3 months after stroke onset. According to motor function recovery at 3 months, patients were divided into good and poor subgroups. The value of multimodal imaging biomarkers of lesion load, lesion volume, white matter integrity, and cortical functional connectivity for motor recovery prediction was investigated in each subgroup.Results. Imaging biomarkers varied depending on recovery pattern. The integrity of the cerebellar tract (P = .005, R2 = .432) was the primary biomarker in the good recovery group. In contrast, the sensory-related corpus callosum tract (P = .026, R2 = .332) and sensory-related functional connectivity (P = .001, R2 = .531) were primary biomarkers in the poor recovery group. A prediction model was proposed by applying each biomarker in the subgroup to patients with different motor evoked potential responses (P < .001, R2 = .853, root mean square error = 5.28).Conclusions. Our results suggest an optimized imaging biomarker model for predicting UE motor recovery after stroke. This model can contribute to individualized management of severe stroke in a clinical setting.

Optimal stimulation site for rTMS to improve motor function: Anatomical hand knob vs. hand motor hotspot.

Repetitive transcranial magnetic stimulation (rTMS) is used to modulate neuronal excitability of the human brain. Distant effects on contralateral corticomotor excitability can be exerted by interhemispheric modulation by low-frequency rTMS on ipsilateral hemisphere. To modulate corticospinal excitability, accurate determination of the stimulation site is important to maximize the effects of rTMS. In the present study, we investigated the difference in the distant effect of 1 Hz rTMS with respect to inducing functional improvement in the non-dominant hand by inhibiting the dominant hemisphere depending on cortical target areas. Ten healthy right-handed volunteers without any neurological disorders were enrolled. The anatomical hand knob (HK) identified from individual magnetic resonance imaging and the transcranial magnetic stimulation (TMS) induced hand motor hotspot (hMHS) by recording motor evoked potentials (MEPs) in the contralateral first dorsal interosseous muscle were determined. All participants underwent three conditions of 1 Hz rTMS on left hemisphere intervention; rTMS application over the HK, rTMS application over the hMHS, and sham-rTMS. Before and after each intervention, all participants undergone motor function assessments with their left hand. The cortical mapping showed that the hMHS was located anteriorly and laterally compared to the HK. Motor function tests showed the most significant improvements after the hMHS stimulation. When we compared the distant effects of target site on corticospinal excitability and motor behavior, delivering 1 Hz rTMS to the hMHS was more effective than delivering it to the HK for improving corticomotor excitability, motor skill, and dexterity. These results suggest that TMS-induced hMHS is an optimal target area to induce distant effect of low-frequency rTMS in motor function.

Comparison of hemodynamic changes after repetitive transcranial magnetic stimulation over the anatomical hand knob and hand motor hotspot: A functional near-infrared spectroscopy study.

BACKGROUND: Low-frequency rTMS can induce upregulation of excitability in the contralateral hemisphere by interhemispheric interaction. OBJECTIVE: The aim of this study was to compare the effects of interhemispheric modulation on hemodynamic changes after applying low-frequency rTMS over the anatomical hand knob (HK) and the hand motor hotspot (hMHS) in the dominant motor cortex. METHODS: Ten healthy right-handed participants without a history of neurological or psychiatric symptoms (five males; 29.8±2.8 years) participated in this single-blind, randomized, cross-over study. rTMS was applied under three conditions over the dominant (left) hemisphere for 20 minutes: 1) 1 Hz rTMS stimulation on the HK (HK-rTMS), 2) 1 Hz rTMS stimulation on the hMHS (hMHS-rTMS), and 3) sham stimulation (Sham-rTMS). For all participants, functional near-infrared spectroscopy (fNIRS) was applied for measurement of cerebral oxyhemoglobin (oxyHb) and deoxyhemoglobin (deoxyHb) concentration over the non-dominant (right) hemisphere during a serial reaction time task (SRTT) with the non-dominant (left) hand before and after each condition. RESULTS: The average coordinates of the hMHS (x = - 39.60 mm, y = - 17.11 mm, z = 66.40 mm) were anterior and lateral to the HK (x = - 36.72 mm, y = - 28.87 mm, z = 56.41 mm). In fNIRS time-series analysis, the integral value of oxyHb wassignificantly increased over the motor cortical region of the non-dominant hemisphere after the hMHS-rTMS compared with Sham-rTMS. The HK-rTMS also showed slight increment of oxyHb concentration but without statistical significance. The SPM group analysis showed greater magnitude of the activity in hMHS-rTMS than that of HK-rTMS after stimulation (p < 0.05). CONCLUSIONS: These results demonstrated an interhemispheric modulation effect of hemodynamic changes by 1 Hz rTMS. The hMHS produced a more robust modulation effect of 1 Hz rTMS on the contralateral hemisphere than did the HK. Therefore, the rTMS can be considered a better stimulation target than the HK.

Modulating Effects of Whole-body Vibration on Cortical Activity and Gait Function in Chronic Stroke Patients.

Whole-body vibration exercise (WBVe) can provide proper somatosensory stimulation and improve muscle strength in stroke patients. This study investigated the effects of WBVe on gait function and cortical activity in patients with chronic stroke. Thirty stroke patients were randomly assigned to either the WBVe or the control group. The WBVe group received the vibration in a half-squat position for 5 minutes at an intensity of 20 Hz. The control group kept the same posture but did not receive the vibration. Cortical activity was investigated using functional near-infrared spectroscopy (fNIRS). Gait function was assessed by a 10-m walk test (10MWT), a timed up and go (TUG) test, a Fugl-Meyer Assessment, and a Tinetti Performance-Oriented Mobility Assessment (TPOMA). In group analysis of the fNIRS data, oxygenated hemoglobin concentration was significantly increased in the ipsilesional supplementary motor area, bilateral sensorimotor cortex, and contralesional prefrontal cortex in the WBVe group compared to the control group (p < 0.05). Functional assessment demonstrated a significant interaction between time and group for the 10MWT and TUG test, suggesting that the WBVe group demonstrated meaningful improvement after intervention (p < 0.05). These results suggested that WBVe modulated the cerebral cortical activities and resulted in improvement of gait function in chronic stroke patients. Trial Registration: ClinicalTrials.gov Identifier: NCT03375346.

Different Brain Connectivity between Responders and Nonresponders to Dual-Mode Noninvasive Brain Stimulation over Bilateral Primary Motor Cortices in Stroke Patients.

Noninvasive brain stimulation (NBS), such as repetitive transcranial magnetic stimulation (rTMS) or transcranial direct current stimulation (tDCS), has been used in stroke patients with motor impairment. NBS can help recovery from brain damage by modulating cortical excitability. However, the efficacy of NBS varies among individuals. To obtain insights of responsiveness to the efficacy of NBS, we investigated characteristic changes of the motor network in responders and nonresponders of NBS over the primary motor cortex (M1). A total of 21 patients with subacute stroke (13 males, mean age 59.6 ± 11.5 years) received NBS in the same manner: 1 Hz rTMS on the contralesional M1 and anodal tDCS on the ipsilesional M1. Participants were classified into responders and nonresponders based on the functional improvement of the affected upper extremity after applying NBS. Twelve age-matched healthy controls (8 males, mean age 56.1 ± 14.3 years) were also recruited. Motor networks were constructed using resting-state functional magnetic resonance imaging. M1 intrahemispheric connectivity, interhemispheric connectivity, and network efficiency were measured to investigate differences in network characteristics between groups. The motor network characteristics were found to differ between both groups. Specifically, M1 intrahemispheric connectivity in responders showed a noticeable imbalance between affected and unaffected hemispheres, which was markedly restored after NBS. The responders also showed greater interhemispheric connectivity and higher efficiency of the motor network than the nonresponders. These results may provide insight on patient-specific NBS treatment based on the brain network characteristics in neurorehabilitation of patients with stroke. This trial is registered with trial registration number NCT03390192.

Modulation of Cortical Activity by High-Frequency Whole-Body Vibration Exercise: An fNIRS Study.

CONTEXT: Whole-body vibration (WBV) has shown many positive effects on the human body in rehabilitation and clinical settings in which vibration has been used to elicit muscle contractions in spastic and paretic muscles. OBJECTIVE: The purpose of this study was to investigate whether WBV exercise (WBVe) differently modulates the cortical activity associated with motor and prefrontal function based on its frequency. METHODS: A total of 18 healthy male adults (mean age: 25.3 [2.4] y) participated in this study and performed WBVe (Galileo Advanced plus; Novotec Medical, Pforzheim, Germany) under 3 different vibration frequency conditions (4-mm amplitude with 10-, 20-, and 27-Hz frequencies) and a control condition (0-mm amplitude with 0-Hz frequency). Each condition consisted of 2 alternating tasks (squatting and standing) every 30 seconds for 5 repetitions. All subjects performed the 4 conditions in a randomized order. MAIN OUTCOME MEASURE: Cortical activation during WBVe was measured by relative changes in oxygenated hemoglobin concentration over the primary motor cortex, premotor cortex, supplementary motor area, and prefrontal and somatosensory cortices using functional near-infrared spectroscopy. RESULTS: Oxygenated hemoglobin concentration was higher during the 27-Hz vibration condition than the control and 10-Hz vibration conditions. Specifically, these changes were pronounced in the bilateral primary motor cortex (P < .05) and right prefrontal cortex (P < .05). In contrast, no significant changes in oxygenated hemoglobin concentration were observed in any of the cortical areas during the 10-Hz vibration condition compared with the control condition. CONCLUSION: This study provides evidence that the motor network and prefrontal cortical areas of healthy adult males can be activated by 27-Hz WBVe. However, WBVe at lower frequencies did not induce significant changes in cortical activation.

Relationship between Serum BDNF Levels and Depressive Mood in Subacute Stroke Patients: A Preliminary Study.

The aim of this preliminary study was to investigate the potential of serum brain-derived neurotrophic factor (BDNF) as a biomarker in poststroke depressive mood in subacute stroke patients. Thirty-eight subacute stroke patients were recruited in this study. All participants underwent the standard rehabilitation program that included 2 h of physical therapy daily and 1 h of occupational therapy five days a week. The rehabilitation period lasted two weeks during the subacute stroke phase. We measured the serum BDNF, proBDNF, and matrix metalloproteinase-9 before and one and two weeks after the standard rehabilitation program. In addition, all participants were assessed using the Geriatric Depression Scale-Short Form (GDS-SF) for depressive mood at three time points. Pearson correlation analysis was performed to determine the relationship between serum BDNF levels and the GDS-SF. The GDS-SF showed significant improvement during the standard rehabilitation program period (p < 0.05). The GDS-SF was significantly correlated with serum BDNF levels at each time point (p < 0.05). These results suggest that serum BDNF may be used as a biomarker for depressive mood in subacute stroke patients. However, further studies with larger study populations are needed to clarify these results.

Differences in motor network dynamics during recovery between supra- and infra-tentorial ischemic strokes.

Most previous stroke studies have been performed in heterogeneous patient populations. Moreover, the brain network might demonstrate different recovery dynamics according to lesion location. In this study, we investigated variation in motor network alterations according to lesion location. Forty patients with subcortical ischemic stroke were enrolled. Patients were divided into two groups: 21 patients with supratentorial stroke (STS) and 19 patients with infratentorial stroke (ITS). All patients underwent resting-state functional magnetic resonance imaging and behavioral assessment at 2 weeks and 3 months poststroke. Twenty-four healthy subjects participated as a control group. To compare altered connectivity between groups, measures used in previous studies to evaluate interhemispheric balance and global network reorganization were investigated and the relationship between network measures and motor functions were examined. Cortico-cerebellar connectivity was also extracted to investigate its relationship with interhemispheric connectivity. In the STS group, measures related to interhemispheric balance were disrupted compared to the control group 2 weeks poststroke, while this was not found in the ITS group. During recovery, measures related to global network reorganization in the STS group and measures related to interhemispheric balance in the ITS group demonstrated significant changes, respectively. Moreover, motor functions were correlated with altered network measures in both groups. There was an interactive relationship between cortico-cerebellar and interhemispheric cortical connectivity only in the ITS group. Different changes in the motor network were observed depending on the location of stroke lesions. These results might originate from differences in the interactions between cortico-cerebellar and interhemispheric connectivity.

Reliable Multivalued Conductance States in TaO x Memristors through Oxygen Plasma-Assisted Electrode Deposition with in Situ-Biased Conductance State Transmission Electron Microscopy Analysis.

Transition metal oxide-based memristors have widely been proposed for applications toward artificial synapses. In general, memristors have two or more electrically switchable stable resistance states that device researchers see as an analogue to the ion channels found in biological synapses. The mechanism behind resistive switching in metal oxides has been divided into electrochemical metallization models and valence change models. The stability of the resistance states in the memristor vary widely depending on: oxide material, electrode material, deposition conditions, film thickness, and programming conditions. So far, it has been extremely challenging to obtain reliable memristors with more than two stable multivalued states along with endurances greater than ∼1000 cycles for each of those states. Using an oxygen plasma-assisted sputter deposition method of noble metal electrodes, we found that the metal-oxide interface could be deposited with substantially lower interface roughness observable at the nanometer scale. This markedly improved device reliability and function, allowing for a demonstration of memristors with four completely distinct levels from ∼6 × 10-6 to ∼4 × 10-8 S that were tested up to 104 cycles per level. Furthermore through a unique in situ transmission electron microscopy study, we were able to verify a redox reaction-type model to be dominant in our samples, leading to the higher degree of electrical state controllability. For solid-state synapse applications, the improvements to electrical properties will lead to simple device structures, with an overall power and area reduction of at least 1000 times when compared to SRAM.

Meso-scale transmission electron microscope tomography applied for wax distribution in toner particles.

Distribution of wax in laser printer toner was observed using an ultra-high-voltage (UHV) and a medium-voltage transmission electron microscope (TEM). As the radius of the wax spans a hundred to greater than a thousand nanometers, its three-dimensional recognition via TEM requires large depth of focus (DOF) for a volumetric specimen. A tomogram with a series of the captured images would allow the determination of their spatial distribution. In this study, bright-field (BF) images acquired with UHV-TEM at a high tilt angle prevented the construction of the tomogram. Conversely, the Z-contrast images acquired by the medium-voltage TEM produced a successful tomogram. The spatial resolution for both is discussed, illustrating that the image degradation was primarily caused by beam divergence of the Z-contrast image and the combination of DOF and chromatic aberration of the BF image from the UHV-TEM.

Spatial distribution of dislocations in relation to a substructure in high-quality GaN film.

The dislocation distribution of high-quality single-crystal gallium nitride (GaN) films grown by the hybrid vapor phase epitaxy was analyzed. This study examined the domain structure of GaN from the dislocation distribution on the macroscale by optical microscopy. The surface structure of GaN consisted of domains with microcolumns as the substructure. The inner domains contained a lower density of dislocations but a large number of these dislocations were observed along the domain boundaries. The existence of a domain boundary structure doubly increased the total dislocation density.

Increased production of bacterial cellulose by Acetobacter sp. V6 in synthetic media under shaking culture conditions.

Acetobacter strains are bacteria that can synthesize cellulose when grown in a complex medium containing glucose. The effect of the components of a synthetic medium on bacterial cellulose (BC) production by a newly isolated Acetobacter sp. V6 in shaking cultures was investigated. BC production was dependent on the presence of MgSO4 x 7H2O and cosubstrates such as ethanol and lactic acid in the medium. The optimal synthetic medium contained 1.5% glucose, 0.2% (NH4)2SO4, 0.3% KH2PO4, 0.3% Na2HPO4 x 12H2O, 0.08% MgSO4 x 7H2O, 0.0005% FeSO4 x 7H2O, 0.0003% H3BO3, 0.00005% nicotinamide, and 0.6% ethanol. A maximum BC concentration of 4.16 g/l was achieved after 8 days of cultivation at 200 rpm. The production of BC by Acetobacter sp. V6 was higher in synthetic medium than complex medium (Hestrin and Schramm medium) traditionally used for Acetobacter strains.