Protective effects of Y-27632 against cisplatin-induced ototoxicity: A zebrafish model Y-27632 and cisplatin-induced ototoxicity.

Cisplatin is an effective chemotherapy agent against various solid malignancies; however, it is associated with irreversible bilateral sensorineural hearing loss, emphasizing the need for drug development to prevent this complication, with the current options being very limited. Rho-associated coiled-coil-containing protein kinase (ROCK) is a serine-threonine protein kinase involved in various cellular processes, including apoptosis regulation. In this study, we used a transgenic zebrafish model (Brn3C: EGFP) in which hair cells within neuromasts are observed in green under fluorescent microscopy without the need for staining. Zebrafish larvae were exposed to cisplatin alone or in combination with various concentrations of Y-27632, a potent ROCK inhibitor. Hair cell counts, apoptosis assessments using the terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling assay, FM1-43FX labeling assay and behavioral analyses (startle response and rheotaxis) were performed to evaluate the protective effects of Y-27632 against cisplatin-induced ototoxicity. Cisplatin treatment reduced the number of hair cells in neuromasts, induced apoptosis, and impaired zebrafish larval behaviors. Y-27632 demonstrated a dose-dependent protective effect against cisplatin-induced hair cell loss and apoptosis. These findings suggest that Y-27632, as a ROCK inhibitor, mitigates cisplatin-induced hair cell loss and associated ototoxicity in zebrafish.

Network Analysis Revealed the Role of Helplessness as a Central Feature Among Late-Life Depressive Symptoms in Patients With Mild Cognitive Impairment and Early Stage Dementia.

OBJECTIVE: It has been reported that depressive symptoms in older adults are different from those in younger adults, especially when accompanied by cognitive decline. However, few studies have investigated the network structure of depressive symptoms in this population. METHODS: The participants consisted of 627 older adults (>60 yr) who were diagnosed with mild cognitive impairment (MCI) or early stage dementia. Among them, 36.7% were male and the mean age was 76.20±7.71 years. The Korean form of Geriatric Depression Scale (KGDS) was used to evaluate their depressive symptoms and network analyses were performed using bootnet R-package to identify the central features among depressive symptoms. RESULTS: Of all the KGDS items, we found that KGDS 2 (often feel helpless) had the highest node strength followed by KGDS 21 (in good spirits), KGDS 14 (not confident at all), and KGDS 15 (cheerful and happy). In terms of node betweenness, KGDS 2 also showed the highest value. The edge weights of edges connected to node KGDS 2 were strongest in KGDS 3 (restless and fidgety) and KGDS 28 (easily get tired). CONCLUSION: In this study, we presented which symptoms are central among the elderly with MCI and early stage dementia. This result not only increases the understanding of depressive symptoms in this group but would also help determine target symptoms in the treatment program.

Temperament Clusters in Patients With Panic Disorder in Relation to Character Maturity.

OBJECTIVE: This study explored whether temperament profiles are associated with psychological functioning and whether character maturity affects this association in patients with panic disorders (PD). METHODS: A total of 270 patients with PD were enrolled in this study. Measurements included the Temperament and Character Inventory-revised-short (TCI-RS), a self-report version of the Panic Disorder Severity Scale (PDSS-SR), Beck Depression Inventory-II (BDI-II), and Spielberger State-Trait Anxiety Inventory (STAI). Cluster analysis was used to define the patients' temperament profiles, and the differences in discrete variables among temperament clusters were calculated using a one-way analysis of variance. An analysis of covariance was conducted to control for the impact of character maturity on psychological functioning among clusters. RESULTS: We identified four temperament clusters of patients with PD. Significant differences in the PDSS-SR, BDI-II, STAI-state, and STAI-trait scores among the four clusters were detected [F(3, 262)=9.16, p<0.001; F(3, 266)=33.78, p<0.001; F(3, 266)=19.12, p<0.001; F(3, 266)=39.46, p<0.001]. However, after controlling for the effect of character maturity, the effect of cluster type was either eliminated or reduced ([STAI-state] cluster type: F(3, 262)=0.94, p>0.05; SD+CO: F(1, 262)=65.95, p<0.001, ηp2 =0.20). CONCLUSION: This study enabled a more comprehensive and integrated understanding of patients by exploring the configuration of all temperament dimensions together rather than each temperament separately. Furthermore, we revealed that depending on the degree of character maturity, the psychological functioning might differ even within the same temperament cluster. These results imply that character maturity can complement inherently vulnerable temperament expression.

Contribution of dysfunctional sleep-related cognitions on insomnia severity: a network perspective.

STUDY OBJECTIVES: Various models of insomnia stress the role of cognitive components, such as dysfunctional sleep-related beliefs, in maintenance and exacerbation of insomnia. This study aimed to use network analysis to identify the particular beliefs that are central and have strong associations with insomnia severity. In addition, we aimed to use a relative importance network to map out predictive pathways between types of dysfunctional beliefs and insomnia severity. METHODS: This study was a retrospective study, with data collected from 219 patients with insomnia. Patients' responses to the Dysfunctional Beliefs about Sleep Scale-16 (DBAS-16) and Insomnia Severity Index (ISI) were collected. All network analyses were performed using R Studio to produce 3 networks: (1) DBAS-16 network, (2) DBAS-16 and ISI network, and (3) relative importance network containing DBAS-16 subscales and ISI. RESULTS: Beliefs reflecting overestimation of negative consequences of sleep (eg, "insomnia is ruining life"), loss of ability (eg, "worry about losing abilities to sleep"), and unpredictability (eg, "can't predict sleep quality") were identified as most central and strongly associated with insomnia severity. Worry/helplessness about insomnia had the largest predictive value on insomnia severity, and also acted as a mediator between other subscales and insomnia severity. CONCLUSIONS: The results of our study suggest that overestimation of negative consequences, loss of ability, and unpredictability are key beliefs that exacerbate and maintain insomnia, thus supporting existing cognitive models of insomnia. CITATION: Cha EJ, Hong S, Kim S, Chung S, Jeon HJ. Contribution of dysfunctional sleep-related cognitions on insomnia severity: a network perspective. J Clin Sleep Med. 2024;20(5):743-751.

Exploring Embryo-Ototoxic Effects: Insights into Deodorant-Induced Hair Cell Damage in Zebrafish.

Our study investigated the embryo-ototoxic effects of deodorant2 (DA2) on zebrafish embryos, which serve as valuable model organisms due to genetic and physiological similarities to humans. We focused on understanding DA2's impact on zebrafish hair cells, which are vital for sensory perception and balance regulation. DA2, provided by the Ministry of Environment, Republic of Korea, was used at 460 µg/mL in dimethyl sulfoxide (DMSO), with a 0.43% DMSO solvent control group. Three experiments, each using 10 zebrafish specimens from each group, showed an initial 13% hair cell count reduction in the DA2-exposed group. Subsequent experiments demonstrated reductions of 37% and 22%, each with one mortality case. Statistical analysis revealed a significant 24% hair cell count reduction in the DA2-exposed group. We also assessed DA2's impact on zebrafish behavior. Although not statistically significant, differences in distances traveled (0.33-0.39, 95% confidence interval: -0.46-1.1, p = 0.2033) and latencies (-0.016-0.018, 95% confidence interval: -0.052-0.021, p = 0.1917) hinted at negative effects. These results highlight DA2's ototoxic properties affecting zebrafish auditory systems and behavior. Further investigation into DA2's effects on aquatic organisms and potential mitigation strategies are essential. These findings contribute to understanding DA2's safety profile, benefiting aquatic ecosystems and human health assessments.

Effects of a blended design of closed-book and open-book examinations on dental students' anxiety and performance.

BACKGROUND: While closed-book examinations (CBEs) have traditionally been implemented in dental education, open-book examinations (OBEs) are being introduced for the purpose of acquiring higher levels of knowledge and promoting long-term memory. This study examines whether it is effective to use a blended assessment of CBEs and OBEs for dental students to reduce test anxiety and enhance academic performance. METHODS: Using a quasi-experimental research method, a blended assessment that combined CBEs in class and OBEs online was designed for a dental course. In 2020, when the pandemic was at its peak, student assessment was ineffective, and the 2020 cohort was omitted for our study; instead, two cohorts of predoctoral dental students (N = 178) enrolled in Restorative Dentistry in the spring semesters of 2019 and 2021 were included in the study. These students were informed about the experimental design, and they provided written consent for data collection, thereby voluntarily participating in the survey. Their self-perceived responses to open-ended survey questions on assessment methods were qualitatively analyzed. RESULTS: There was no significant difference in test anxiety between the CBEs and OBEs at the p-values of 0.001 in all items. Traditional and blended assessment showed a similar trend of lower scores in midterm exams compared to higher scores in final exams, thus discriminating against students' performances. In particular, a low-achieving group was better predicted by a blended assessment. An analysis of the students' self-perceived responses produced highly topical themes, including exam burden, learning effects, and fairness issues. CONCLUSIONS: This study confirmed the feasibility of blended assessment that can be implemented in online and in-person educational environments. Moreover, it can be used as the groundwork to develop new models of assessment in dental education.

Do the oral health-related quality of life and the magnitude of its association with malocclusion differ by developmental stages of schoolchildren?

INTRODUCTION: As schoolchildren go through multiple developmental periods, their oral health-related quality of life (OHRQOL) level may change over time. It is important to understand the association between malocclusion and OHRQOL in treatment planning for schoolchildren. This study aimed to examine OHRQOL and its association with malocclusion among schoolchildren at different developmental stages. METHODS: Data from a representative sample of 2010 Korean schoolchildren aged 7-17 years were analyzed. The participants completed a self-administered questionnaire, and their orthodontic treatment needs were assessed by a dentist. OHRQOL was measured using the Child Oral Health Impact Profile (COHIP). Higher COHIP scores indicate better OHRQOL. RESULTS: The mean age of the sample was 11.9 ± 2.5 years, and slightly more boys (53.0%) were in the sample. The total COHIP score (P <0.001) and 4 subscale scores (P <0.01) were lower in older age groups. Compared with those who needed orthodontic treatment, children with no and borderline orthodontic treatment needs were 1.9 times (odds ratio [OR], 1.9; 95% confidence interval, 1.5-2.4) and 1.5 times (OR, 1.5; 95% confidence interval, 1.1-2.1) more likely to have a higher COHIP score, respectively. The degree of association varied across age groups: a larger effect was found in the oldest group (aged 15 years) (OR, 4.1) than in younger groups (ORs, 1.5-2.2). CONCLUSIONS: OHRQOL and the magnitude of its association with malocclusion varied by age among schoolchildren, particularly in functional and social-emotional well-being. Given the variation, schoolchildren's developmental stages should be considered in treatment planning.

Association between Heart Rate Variability Indices and Depressed Mood in Patients with Panic Disorder.

Objective: Heart rate variability (HRV) reflects the regulation of the autonomic nervous system. Panic disorder is highly associated with autonomic dysfunction, and is often accompanied by depression. The aim of this study is to determine the association between depression and HRV indices in patients with panic disorder. Methods: A total of 110 outpatients diagnosed with panic disorder participated in this study. The medical records of patients with panic disorder who visited the outpatient clinic of Konkuk University Hospital between December 2018 and March 2020 were retrospectively reviewed. Measurements used in this study include the Panic Disorder Severity Scale-Self Report, Beck Depression Inventory (BDI-II), Insomnia Severity Index, and HRV. Patients were divided into depressive and non-depressive groups based on their BDI-II scores. The association between HRV indices and depressive symptoms was statistically analyzed. Results: The low frequency/high frequency (LF/HF) ratio was reduced in patients with depression (mean = -0.095, p = 0.004 in the above moderate depressive group, mean = -0.120, p = 0.020 in the severe depressive group). Significant correlations were found between depressive symptoms and standard deviation of NN interval (SDNN) (ms) (-0.19, p = 0.044), very low frequency (VLF) (ms2/Hz) (-0.22, p = 0.021), LF (-0.25, p = 0.008), HF (-0.19, p = 0.043), and LF/HF (-0.25, p = 0.009). Multiple linear regression analysis showed that BDI predicted SDNN (ms), VLF (ms2/Hz), LF, HF, and LF/HF. Conclusion: We confirmed that the LF/HF ratio decreases when depression is accompanied by panic disorder. HRV indices may be useful markers for detecting depressive symptoms in patients with panic disorder.

Differences in temperament and character inventory (TCI) profile between suicidal and nonsuicidal psychiatric outpatients.

The purpose of this study was to identify personality traits associated with suicide attempt in a clinical sample. Temperament and character inventory (TCI) profiles of 759 patients who met the inclusion criteria among 1000 randomly selected hospital records collected. Of these 759 patients, 103 had a history of at least 1 actual suicide attempt (suicidal group) whereas 656 had no such history (nonsuicidal group). The suicidal group showed higher scores of novelty seeking (mean ± SD: 36.1 ± 1.2 vs 33.3 ± 0.5; P = .026) and harm avoidance (57.1 ± 1.5 vs 53.0 ± 0.6; P = .01) but lower scores of self-directedness (27.5 ± 1.5 vs 34.4 ± 0.6; P < .001) than the nonsuicidal group. Higher novelty seeking (OR [95% CI]: 1.031 [1.008-1.054]; P = .007) and lower self-directedness: 0.955 [0.927-0.983]; P = .002 were also associated with suicide attempts in the analysis of 7 personality scales. These findings suggest that patients who attempt suicide differ from nonattempters in terms of personality traits, especially in novelty seeking (NS), harm avoidance (HA), and self-directedness (SD). It is noteworthy that this study contains data from actual visits to the emergency room to evaluate suicide attempts. Abbreviations: CO = cooperativeness, Ha = harm avoidance, NS = novelty seeking, PS = persistence, RD = reward dependence, SD = self-directedness, ST = self-transcendence, TCI = temperament and character inventory.

A network analysis of panic symptoms in relation to depression and anxiety sensitivity in patients with panic disorder.

BACKGROUND: Panic disorder is an anxiety disorder presenting panic attacks as a defining feature, with cognitive and behavioral symptoms that are associated with the panic attacks. Recently, the use of network analysis is increasing to determine the symptoms and mutual reinforcing patterns that conceptualize a mental disorder. This study aimed to improve our understanding of panic disorder by estimating a network structure of its symptoms. METHOD: Data from 257 patients diagnosed with panic disorder who visited the outpatient psychiatric clinic from 2018 to 2020 were collected. Panic attacks, cognitive and behavioral symptoms concerning the panic attacks, depression, and anxiety sensitivity dimensions such as fear of cardiovascular symptom, fear of publicly observable anxiety reaction, fear of respiratory symptom, and fear of cognitive dyscontrol were computed. Bootstrapping was applied to estimate the 95% confidence intervals for each edges. RESULTS: The resulting network indicated fear of cognitive dyscontrol, impairment in social functioning, phobic avoidance of situations, and panic-focused anticipatory anxiety to be central symptoms in panic disorder. Interestingly, panic attacks and distress during panic did not emerge as central. Depression was strongly linked to fear of cognitive dyscontrol. LIMITATIONS: The sample size limits network comparison tests between those with comorbid depression or agoraphobia. In addition, the cross-sectional design limits the opportunity to draw causal conclusions regarding the symptoms. CONCLUSIONS: These results have implications in etiology of panic disorder and support the efficaciousness of Cognitive Behavioral Therapy. Future network analyses may employ longitudinal designs to investigate causal relationships between the symptoms.

Viral Shrimp Diseases Listed by the OIE: A Review.

Shrimp is one of the most valuable aquaculture species globally, and the most internationally traded seafood product. Consequently, shrimp aquaculture practices have received increasing attention due to their high value and levels of demand, and this has contributed to economic growth in many developing countries. The global production of shrimp reached approximately 6.5 million t in 2019 and the shrimp aquaculture industry has consequently become a large-scale operation. However, the expansion of shrimp aquaculture has also been accompanied by various disease outbreaks, leading to large losses in shrimp production. Among the diseases, there are various viral diseases which can cause serious damage when compared to bacterial and fungi-based illness. In addition, new viral diseases occur rapidly, and existing diseases can evolve into new types. To address this, the review presented here will provide information on the DNA and RNA of shrimp viral diseases that have been designated by the World Organization for Animal Health and identify the latest shrimp disease trends.

The Mediating Effect of Insomnia on the Relationship between Panic Symptoms and Depression in Patients with Panic Disorder.

BACKGROUND: This study aimed to determine if sleep disturbances may mediate the relationship between panic symptoms and depression in patients with panic disorder (PD). METHODS: Electronic medical records were retrospectively reviewed for 110 consecutive patients with diagnosed PD in an outpatient clinic between October 2018 and December 2019. Measurements include the PD Severity Scale, Beck Depression Inventory-II (BDI-II) and Insomnia Severity Index (ISI). Statistical analyses were performed to assess any potential relationship between PD, insomnia and depression. RESULTS: Of the PD patients, 88 (80%) and 89 (80.9%) had comorbid depression (BDI-II ≥ 14) and insomnia (Korean version of the ISI ≥ 8), respectively. In a mediation model using insomnia as the mediating variable, the total effect of panic symptom severity on depression was significant (t = 7.23, P < 0.001). There were significant effects of panic symptoms on insomnia (t = 4.62, P < 0.001) and of insomnia on depression (t = 6.69, P < 0.001). The main effect of panic symptom severity on depression was also significant, after controlling for the effect of insomnia (t = 5.10, P < 0.001), suggesting partial mediation. CONCLUSION: Both depressive symptoms and insomnia are common in patients with PD and depression was partially mediated by insomnia in these patients. These results suggest that an intervention for insomnia in patients with PD might help prevent the development of depression.

Distributed Interactive Visualization Using GPU-Optimized Spark.

With the advent of advances in imaging and computing technologies, large-scale data acquisition and processing have become commonplace in many science and engineering disciplines. Conventional workflows for large-scale data processing usually rely on in-house or commercial software that are designed for domain-specific computing tasks. Recent advances in MapReduce, which was originally developed for batch processing textual data via a simplified programming model of the map and reduce functions, have expanded its applications to more general tasks in big-data processing, such as scientific computing, and biomedical image processing. However, as shown in previous work, volume rendering and visualization using MapReduce is still considered challenging and impractical owing to the disk-based, batch-processing nature of its computing model. In this article, contrary to this common belief, we show that the MapReduce computing model can be effectively used for interactive visualization. Our proposed system is a novel extension of Spark, one of the most popular open-source MapReduce frameworks, which offers GPU-accelerated MapReduce computing. To minimize CPU-GPU communication and overcome slow, disk-based shuffle performance, the proposed system supports GPU in-memory caching and MPI-based direct communication between compute nodes. To allow for GPU-accelerated in-situ visualization using raster graphics in Spark, we leveraged the CUDA-OpenGL interoperability, resulting in faster processing speeds by several orders of magnitude compared to conventional MapReduce systems. We demonstrate the performance of our system via several volume processing and visualization tasks, such as direct volume rendering, iso-surface extraction, and numerical simulations with in-situ visualization.

Level of professional ethics awareness and medical ethics competency of dental hygienists and dental hygiene students: the need to add ethics items to the Korean Dental Hygienist Licensing Examination

PURPOSE: This study aimed to evaluate the level of professional ethics awareness and medical ethics competency in order to assess the potential need for ethics items to be included on the Korean Dental Hygienist Licensing Examination. METHODS: In total, 358 clinical dental hygienists and dental hygiene students completed a structured questionnaire to evaluate their level of ethical awareness and medical ethics competency. The sub-factors of medical ethics were classified into relationships with patients, medical and social relations, and individual specialized fields. RESULTS: Only 32.1% of participants indicated that they had taken a course on professional ethics in the university curriculum, but 95.2% of respondents considered professional ethics to be important. The overall score for medical ethics competency was average (3.37 out of 5). The score for relationships with patients was 3.75 points, followed by medical and social relations (3.19 points) and individual specialized fields (3.16 points). The level of professional ethics awareness was higher among participants who had taken a course on professional ethics than among those who had not done so or who did not remember whether they had done so. CONCLUSION: Dental hygienists were aware of the importance of professional ethics, but their medical ethics competency was moderate. Therefore, medical ethics should be treated as a required subject in the university curriculum, and medical ethics competency evaluations should be strengthened by adding ethics items to the Korean Dental Hygienist Licensing Examination.

Lasso Proteins: Modular Design, Cellular Synthesis, and Topological Transformation.

Entangled proteins have attracted significant research interest. Herein, we report the first rationally designed lasso proteins, or protein [1]rotaxanes, by using a p53dim-entwined dimer for intramolecular entanglement and a SpyTag-SpyCatcher reaction for side-chain ring closure. The lasso structures were confirmed by proteolytic digestion, mutation, NMR spectrometry, and controlled ligation. Their dynamic properties were probed by experiments such as end-capping, proteolytic digestion, and heating/cooling. As a versatile topological intermediate, a lasso protein could be converted to a rotaxane, a heterocatenane, and a "slide-ring" network. Being entirely genetically encoded, this robust and modular lasso-protein motif is a valuable addition to the topological protein repertoire and a promising candidate for protein-based biomaterials.

Larkinella roseus sp. nov., a species of the family Cytophagaceae isolated from beach soil.

The taxonomic position of bacterial strain, designated 15J16-1T3AT, recovered from a soil sample was established using a polyphasic approach. Phylogenic analysis based on the 16S rRNA gene sequence showed that strain 15J16-1T3AT belonged to the family Cytophagaceae, phylum Bacteroidetes, and was most closely related to 'Larkinella harenae' 15J9-9 (95.9% similarity), Larkinella ripae 15J11-11T (95.6%), Larkinella bovis M2TB15T (94.7%), Larkinella arboricola Z0532T (93.9%), and Larkinella insperata LMG 22510T (93.5%). Cells were rod-shaped, Gram-stain-negative, aerobic, and nonmotile. The isolate grew on NA, R2A, TSA, but not on LB agar. The strain was able to grow at temperature range from 10°C to 30°C with an optimum at 25°C and pH 6-8. Menaquinone MK-7 was the predominant respiratory quinone. The major cellular fatty acids comprised C16:1ω5c (48.6%) and C15:0 iso (24.1%). Phosphatidylethanolamine, phosphatidylserine, and an unidentified lipid were the major polar lipids. The G + C content of the genomic DNA was 49.5 mol%. Strain 15J16-1T3AT could be distinguished from its closest phylogenetic neighbors based on its phenotypic, genotypic, and chemotaxonomic features. Therefore, the isolate is considered to represent a novel species in the genus Larkinella, for which the name Larkinella roseus sp. nov. is proposed. The type strain is 15J16-1T3AT (= KCTC 52004T = JCM 31991T).

Validation of a Korean version of the Child Oral Health Impact Profile (COHIP) among 8- to 15-year-old school children.

OBJECTIVES: The purpose of this study was to assess reliability, discriminant validity, and convergent validity of the Oral Health Impact Profile (COHIP) Korean version in a representative community sample of 8- to 15-year-old Korean children. METHODS: A Korean version of COHIP was developed according to the standard procedure of cross-cultural adaptation of self-reported instruments. A representative community sample of 2236 schoolchildren was selected by cluster sampling method. RESULTS: Mean age of the participants was 11.8 years. Mean and median of the overall COHIP score were 103.3 (SD 13.3) and 106, respectively. Internal reliability and retest reliability were excellent with Chronbach's alpha 0.88 and intraclass correlation coefficient 0.88. Face validity was confirmed with 98% of participants reporting the COHIP questionnaire was easy to answer. Nonclinical factors such as self-rated oral health or satisfaction with oral health were significantly related with overall COHIP score and five subscale scores (P < 0.001) in a consistent manner. Children with carious permanent teeth and with orthodontic treatment need had highly significantly lower overall COHIP score (P < 0.01). CONCLUSION: The Korean version of the COHIP was successfully developed. The internal reliability, retest reliability, face validity, discriminant validity, and convergent validity of the COHIP Korean version were confirmed.

Senile Cardiac Calcification Syndrome: A Rare Case of Extensive Calcification of Left Ventricular Papillary Muscle.

Extensive papillary muscle calcification is uncommon and only scarce literature about causes and the clinical significance is available, whereas small calcific deposits are common findings in elderly people and are located most commonly at the apex. Papillary muscle calcification has been associated with coronary artery disease, dilated cardiomyopathy, mitral valve disease, hypercalcemia, and increased calcium phosphate product in end stage renal disease. We reported a rare case of extensive calcification of anterolateral papillary muscle diagnosed by echocardiography and multidetector computed tomography.

A Case of Cardiac Amyloidosis Initially Misdiagnosed as Syndrome X.

Cardiac infiltration of amyloid fibril results in progressive cardiomyopathy with a grave prognosis and results in cardiac diseases such as congestive heart disease, cardiomyopathy, valvular heart disease, and arrhythmias. We present a rare case of cardiac amyloidosis initially misdiagnosed as syndrome X in which recurrent chest pain and progressive heart failure could be managed finally by heart transplantation.