Adoption of AI in Oncological Imaging: Ethical, Regulatory, and Medical-Legal Challenges.

Artificial Intelligence (AI) algorithms have shown great promise in oncological imaging, outperforming or matching radiologists in retrospective studies, signifying their potential for advanced screening capabilities. These AI tools offer valuable support to radiologists, assisting them in critical tasks such as prioritizing reporting, early cancer detection, and precise measurements, thereby bolstering clinical decision-making. With the healthcare landscape witnessing a surge in imaging requests and a decline in available radiologists, the integration of AI has become increasingly appealing. By streamlining workflow efficiency and enhancing patient care, AI presents a transformative solution to the challenges faced by oncological imaging practices. Nevertheless, successful AI integration necessitates navigating various ethical, regulatory, and medical-legal challenges. This review endeavors to provide a comprehensive overview of these obstacles, aiming to foster a responsible and effective implementation of AI in oncological imaging.

Radiomics and Artificial Intelligence in Renal Lesion Assessment.

Radiomics, the extraction and analysis of quantitative features from medical images, has emerged as a promising field in radiology with the potential to revolutionize the diagnosis and management of renal lesions. This comprehensive review explores the radiomics workflow, including image acquisition, feature extraction, selection, and classification, and highlights its application in differentiating between benign and malignant renal lesions. The integration of radiomics with artificial intelligence (AI) techniques, such as machine learning and deep learning, can help patients' management and allow the planning of the appropriate treatments. AI models have shown remarkable accuracy in predicting tumor aggressiveness, treatment response, and patient outcomes. This review provides insights into the current state of radiomics and AI in renal lesion assessment and outlines future directions for research in this rapidly evolving field.

Artificial Intelligence in Lung Cancer Imaging: From Data to Therapy.

Lung cancer remains a global health challenge, leading to substantial morbidity and mortality. While prevention and early detection strategies have improved, the need for precise diagnosis, prognosis, and treatment remains crucial. In this comprehensive review article, we explore the role of artificial intelligence (AI) in reshaping the management of lung cancer. AI may have different potential applications in lung cancer characterization and outcome prediction. Manual segmentation is a time-consuming task, with high inter-observer variability, that can be replaced by AI-based approaches, including deep learning models such as U-Net, BCDU-Net, and others, to quantify lung nodules and cancers objectively and to extract radiomics features for the characterization of the tissue. AI models have also demonstrated their ability to predict treatment responses, such as immunotherapy and targeted therapy, by integrating radiomic features with clinical data. Additionally, AI-based prognostic models have been developed to identify patients at higher risk and personalize treatment strategies. In conclusion, this review article provides a comprehensive overview of the current state of AI applications in lung cancer management, spanning from segmentation and virtual biopsy to outcome prediction. The evolving role of AI in improving the precision and effectiveness of lung cancer diagnosis and treatment underscores its potential to significantly impact clinical practice and patient outcomes.

How do patients perceive the AI-radiologists interaction? Results of a survey on 2119 responders.

PURPOSE: In this study we investigate how patients perceive the interaction between artificial intelligence (AI) and radiologists by designing a survey. METHOD: We created a survey focused on the application of Artificial Intelligence in radiology which consisted of 20 questions distributed in three sections:Only completed questionnaires were considered for analysis. RESULTS: 2119 subjects completed the survey. Among them, 1216 respondents were over 60 years old, showing interest in AI even though they were not digital natives. Although >45% of the respondents reported a high level of education, only 3% said they were AI experts. 87% of respondents favored using AI to support diagnosis but would like to be informed. Only 10% would consult another specialist if their doctor used AI support. Most respondents (76%) said they would not feel comfortable if the diagnosis was made by the AI alone, highlighting the importance of the physician's role in the emotional management of the patient. Finally, 36% of respondents were willing to discuss the topic further in a focus group. CONCLUSION: Patients' perception of the use of AI in radiology was positive, although still strictly linked to the supervision of the radiologist. Respondents showed interest and willingness to learn more about AI in the medical field, confirming how patients' confidence in AI technology and its acceptance is central to its widespread use in clinical practice.

Amplifying the Effects of Contrast Agents on Magnetic Resonance Images Using a Deep Learning Method Trained on Synthetic Data.

OBJECTIVES: Artificial intelligence (AI) methods can be applied to enhance contrast in diagnostic images beyond that attainable with the standard doses of contrast agents (CAs) normally used in the clinic, thus potentially increasing diagnostic power and sensitivity. Deep learning-based AI relies on training data sets, which should be sufficiently large and diverse to effectively adjust network parameters, avoid biases, and enable generalization of the outcome. However, large sets of diagnostic images acquired at doses of CA outside the standard-of-care are not commonly available. Here, we propose a method to generate synthetic data sets to train an "AI agent" designed to amplify the effects of CAs in magnetic resonance (MR) images. The method was fine-tuned and validated in a preclinical study in a murine model of brain glioma, and extended to a large, retrospective clinical human data set. MATERIALS AND METHODS: A physical model was applied to simulate different levels of MR contrast from a gadolinium-based CA. The simulated data were used to train a neural network that predicts image contrast at higher doses. A preclinical MR study at multiple CA doses in a rat model of glioma was performed to tune model parameters and to assess fidelity of the virtual contrast images against ground-truth MR and histological data. Two different scanners (3 T and 7 T, respectively) were used to assess the effects of field strength. The approach was then applied to a retrospective clinical study comprising 1990 examinations in patients affected by a variety of brain diseases, including glioma, multiple sclerosis, and metastatic cancer. Images were evaluated in terms of contrast-to-noise ratio and lesion-to-brain ratio, and qualitative scores. RESULTS: In the preclinical study, virtual double-dose images showed high degrees of similarity to experimental double-dose images for both peak signal-to-noise ratio and structural similarity index (29.49 dB and 0.914 dB at 7 T, respectively, and 31.32 dB and 0.942 dB at 3 T) and significant improvement over standard contrast dose (ie, 0.1 mmol Gd/kg) images at both field strengths. In the clinical study, contrast-to-noise ratio and lesion-to-brain ratio increased by an average 155% and 34% in virtual contrast images compared with standard-dose images. Blind scoring of AI-enhanced images by 2 neuroradiologists showed significantly better sensitivity to small brain lesions compared with standard-dose images (4.46/5 vs 3.51/5). CONCLUSIONS: Synthetic data generated by a physical model of contrast enhancement provided effective training for a deep learning model for contrast amplification. Contrast above that attainable at standard doses of gadolinium-based CA can be generated through this approach, with significant advantages in the detection of small low-enhancing brain lesions.

Chest X-ray in Emergency Radiology: What Artificial Intelligence Applications Are Available?

Due to its widespread availability, low cost, feasibility at the patient's bedside and accessibility even in low-resource settings, chest X-ray is one of the most requested examinations in radiology departments. Whilst it provides essential information on thoracic pathology, it can be difficult to interpret and is prone to diagnostic errors, particularly in the emergency setting. The increasing availability of large chest X-ray datasets has allowed the development of reliable Artificial Intelligence (AI) tools to help radiologists in everyday clinical practice. AI integration into the diagnostic workflow would benefit patients, radiologists, and healthcare systems in terms of improved and standardized reporting accuracy, quicker diagnosis, more efficient management, and appropriateness of the therapy. This review article aims to provide an overview of the applications of AI for chest X-rays in the emergency setting, emphasizing the detection and evaluation of pneumothorax, pneumonia, heart failure, and pleural effusion.

Artificial Intelligence in Emergency Radiology: Where Are We Going?

Emergency Radiology is a unique branch of imaging, as rapidity in the diagnosis and management of different pathologies is essential to saving patients' lives. Artificial Intelligence (AI) has many potential applications in emergency radiology: firstly, image acquisition can be facilitated by reducing acquisition times through automatic positioning and minimizing artifacts with AI-based reconstruction systems to optimize image quality, even in critical patients; secondly, it enables an efficient workflow (AI algorithms integrated with RIS-PACS workflow), by analyzing the characteristics and images of patients, detecting high-priority examinations and patients with emergent critical findings. Different machine and deep learning algorithms have been trained for the automated detection of different types of emergency disorders (e.g., intracranial hemorrhage, bone fractures, pneumonia), to help radiologists to detect relevant findings. AI-based smart reporting, summarizing patients' clinical data, and analyzing the grading of the imaging abnormalities, can provide an objective indicator of the disease's severity, resulting in quick and optimized treatment planning. In this review, we provide an overview of the different AI tools available in emergency radiology, to keep radiologists up to date on the current technological evolution in this field.

Artificial Intelligence in Lung Cancer Imaging: Unfolding the Future.

Lung cancer is one of the malignancies with higher morbidity and mortality. Imaging plays an essential role in each phase of lung cancer management, from detection to assessment of response to treatment. The development of imaging-based artificial intelligence (AI) models has the potential to play a key role in early detection and customized treatment planning. Computer-aided detection of lung nodules in screening programs has revolutionized the early detection of the disease. Moreover, the possibility to use AI approaches to identify patients at risk of developing lung cancer during their life can help a more targeted screening program. The combination of imaging features and clinical and laboratory data through AI models is giving promising results in the prediction of patients' outcomes, response to specific therapies, and risk for toxic reaction development. In this review, we provide an overview of the main imaging AI-based tools in lung cancer imaging, including automated lesion detection, characterization, segmentation, prediction of outcome, and treatment response to provide radiologists and clinicians with the foundation for these applications in a clinical scenario.

Development and Validation of an AI-driven Mammographic Breast Density Classification Tool Based on Radiologist Consensus.

Mammographic breast density (BD) is commonly visually assessed using the Breast Imaging Reporting and Data System (BI-RADS) four-category scale. To overcome inter- and intraobserver variability of visual assessment, the authors retrospectively developed and externally validated a software for BD classification based on convolutional neural networks from mammograms obtained between 2017 and 2020. The tool was trained using the majority BD category determined by seven board-certified radiologists who independently visually assessed 760 mediolateral oblique (MLO) images in 380 women (mean age, 57 years ± 6 [SD]) from center 1; this process mimicked training from a consensus of several human readers. External validation of the model was performed by the three radiologists whose BD assessment was closest to the majority (consensus) of the initial seven on a dataset of 384 MLO images in 197 women (mean age, 56 years ± 13) obtained from center 2. The model achieved an accuracy of 89.3% in distinguishing BI-RADS a or b (nondense breasts) versus c or d (dense breasts) categories, with an agreement of 90.4% (178 of 197 mammograms) and a reliability of 0.807 (Cohen κ) compared with the mode of the three readers. This study demonstrates accuracy and reliability of a fully automated software for BD classification. Keywords: Mammography, Breast, Convolutional Neural Network (CNN), Deep Learning Algorithms, Machine Learning Algorithms Supplemental material is available for this article. © RSNA, 2022.

Evolutionary conservation of a regulative pathway of erythropoiesis in Poikilothermic vertebrates.

Apoptosis, programmed cell death, plays a central role in haematopoiesis. Mature erythrocytes of non-mammalian vertebrates maintain a permanent nucleus; these cells can undergo apoptosis (eryptosis), as do other somatic cells of a given non-mammalian vertebrate. In this study, we have investigated the expression and subcellular distribution of Bcl-2, Bcl-XL and Bax proteins in the maturation phases and after X-ray irradiation of nucleated erythrocytes of Torpedo marmorata and Caretta caretta and the effect of X-ray irradiation on nucleated circulating erythrocytes of Torpedo marmorata. The cellular distribution of proteins was detected in erythrocytes by using immunocytochemistry at light microscopy and immunoelectron microscopy. The electrophoretic separation and immunoblotting of pro- and anti-apoptotic proteins of immature and mature erythroid cells was performed too, after X-ray irradiation of torpedoes. The results of the immunocytochemical analyses show an increase, in the expression level of Bax in mature as compared to young erythrocytes and a corresponding decrease of Bcl-2 and Bcl-XL. This maturation pattern of Bax, Bcl-2 and Bcl-XL was abrogated in X-ray irradiated torpedo erythrocytes. On the basis of these observations, Bax, Bcl-2 and Bcl-XL seems to play a role in the erythropoiesis of Torpedo marmorata Risso and in Caretta caretta. In conclusion, the same apoptotic proteins of somatic cells appear to be conserved in circulating nucleated erythrocytes thus suggesting to play a role in the maturation of these cells.

AIforCOVID: Predicting the clinical outcomes in patients with COVID-19 applying AI to chest-X-rays. An Italian multicentre study.

Recent epidemiological data report that worldwide more than 53 million people have been infected by SARS-CoV-2, resulting in 1.3 million deaths. The disease has been spreading very rapidly and few months after the identification of the first infected, shortage of hospital resources quickly became a problem. In this work we investigate whether artificial intelligence working with chest X-ray (CXR) scans and clinical data can be used as a possible tool for the early identification of patients at risk of severe outcome, like intensive care or death. Indeed, further to induce lower radiation dose than computed tomography (CT), CXR is a simpler and faster radiological technique, being also more widespread. In this respect, we present three approaches that use features extracted from CXR images, either handcrafted or automatically learnt by convolutional neuronal networks, which are then integrated with the clinical data. As a further contribution, this work introduces a repository that collects data from 820 patients enrolled in six Italian hospitals in spring 2020 during the first COVID-19 emergency. The dataset includes CXR images, several clinical attributes and clinical outcomes. Exhaustive evaluation shows promising performance both in 10-fold and leave-one-centre-out cross-validation, suggesting that clinical data and images have the potential to provide useful information for the management of patients and hospital resources.

The allosteric protein interactions in the proton-motive function of mammalian redox enzymes of the respiratory chain.

Insight into mammalian respiratory complexes defines the role of allosteric protein interactions in their proton-motive activity. In cytochrome c oxidase (CxIV) conformational change of subunit I, caused by O2 binding to heme a32+-CuB+ and reduction, and stereochemical transitions coupled to oxidation/reduction of heme a and CuA, combined with electrostatic effects, determine the proton pumping activity. In ubiquinone-cytochrome c oxidoreductase (CxIII) conformational movement of Fe-S protein between cytochromes b and c1 is the key element of the proton-motive activity. In NADH-ubiquinone oxidoreductase (CxI) ubiquinone binding and reduction result in conformational changes of subunits in the quinone reaction structure which initiate proton pumping.

Prevalence of interstitial pneumonia suggestive of COVID-19 at 18F-FDG PET/CT in oncological asymptomatic patients in a high prevalence country during pandemic period: a national multi-centric retrospective study.

PURPOSE: To assess the presence and pattern of incidental interstitial lung alterations suspicious of COVID-19 on fluorine-18-fluorodeoxyglucose positron emission tomography (PET)/computed tomography (CT) ([18F]FDG PET/CT) in asymptomatic oncological patients during the period of active COVID-19 in a country with high prevalence of the virus. METHODS: This is a multi-center retrospective observational study involving 59 Italian centers. We retrospectively reviewed the prevalence of interstitial pneumonia detected during the COVID period (between March 16 and 27, 2020) and compared to a pre-COVID period (January-February 2020) and a control time (in 2019). The diagnosis of interstitial pneumonia was done considering lung alterations of CT of PET. RESULTS: Overall, [18F]FDG PET/CT was performed on 4008 patients in the COVID period, 19,267 in the pre-COVID period, and 5513 in the control period. The rate of interstitial pneumonia suspicious for COVID-19 was significantly higher during the COVID period (7.1%) compared with that found in the pre-COVID (5.35%) and control periods (5.15%) (p < 0.001). Instead, no significant difference among pre-COVID and control periods was present. The prevalence of interstitial pneumonia detected at PET/CT was directly associated with geographic virus diffusion, with the higher rate in Northern Italy. Among 284 interstitial pneumonia detected during COVID period, 169 (59%) were FDG-avid (average SUVmax of 4.1). CONCLUSIONS: A significant increase of interstitial pneumonia incidentally detected with [18F]FDG PET/CT has been demonstrated during the COVID-19 pandemic. A majority of interstitial pneumonia were FDG-avid. Our results underlined the importance of paying attention to incidental CT findings of pneumonia detected at PET/CT, and these reports might help to recognize early COVID-19 cases guiding the subsequent management.

Comparison of Transfer Learning and Conventional Machine Learning Applied to Structural Brain MRI for the Early Diagnosis and Prognosis of Alzheimer's Disease.

Alzheimer's Disease (AD) is the most common neurodegenerative disease, with 10% prevalence in the elder population. Conventional Machine Learning (ML) was proven effective in supporting the diagnosis of AD, while very few studies investigated the performance of deep learning and transfer learning in this complex task. In this paper, we evaluated the potential of ensemble transfer-learning techniques, pretrained on generic images and then transferred to structural brain MRI, for the early diagnosis and prognosis of AD, with respect to a fusion of conventional-ML approaches based on Support Vector Machine directly applied to structural brain MRI. Specifically, more than 600 subjects were obtained from the ADNI repository, including AD, Mild Cognitive Impaired converting to AD (MCIc), Mild Cognitive Impaired not converting to AD (MCInc), and cognitively-normal (CN) subjects. We used T1-weighted cerebral-MRI studies to train: (1) an ensemble of five transfer-learning architectures pretrained on generic images; (2) a 3D Convolutional Neutral Network (CNN) trained from scratch on MRI volumes; and (3) a fusion of two conventional-ML classifiers derived from different feature extraction/selection techniques coupled to SVM. The AD-vs-CN, MCIc-vs-CN, MCIc-vs-MCInc comparisons were investigated. The ensemble transfer-learning approach was able to effectively discriminate AD from CN with 90.2% AUC, MCIc from CN with 83.2% AUC, and MCIc from MCInc with 70.6% AUC, showing comparable or slightly lower results with the fusion of conventional-ML systems (AD from CN with 93.1% AUC, MCIc from CN with 89.6% AUC, and MCIc from MCInc with AUC in the range of 69.1-73.3%). The deep-learning network trained from scratch obtained lower performance than either the fusion of conventional-ML systems and the ensemble transfer-learning, due to the limited sample of images used for training. These results open new prospective on the use of transfer learning combined with neuroimages for the automatic early diagnosis and prognosis of AD, even if pretrained on generic images.

Open 1.0-T versus closed 1.5-T cardiac MR: Image quality assessment.

PURPOSE: The aim of this paper was to compare the open 1-T (O-1T) versus the closed 1.5-T (C-1.5T) cardiac magnetic resonance (MR). PATIENTS/METHODS: The MR examinations of two concurrent cohorts (each including 100 subjects) of patients with suspected or known cardiac disease were reviewed. Such examinations were obtained using O-1T or C-1.5T MRI. The bright-blood cine, T1-weighted (T1), T2-weighed short-tau inversion recovery (T2-STIR), late gadolinium enhancement (LGE) sequences were performed. Signal-to-noise ratio of blood (SNRb) or myocardium (SNRm), and contrast-to-noise ratio of myocardium (CNRm) were calculated. Subjective image quality (SIQ) of each sequence was graded as 0 = poor, 1 = intermediate, or 2 = optimal. Each examination was considered as diagnostic when the report answered the clinical question. RESULTS: C-1.5T was better than O-1T on cine for SNRb(median 172 versus 452), SNRm(71 versus 160) and CNRm (107 versus 265) and on T2-STIR for SNRb(10 versus 29), SNRm(74 versus 261) and CNRm(-67 versus -233)(P < 0.001). On LGE, SNRm was higher with O-1T than for C-1.5T (312 versus 79, P < 0.001) while CNR was lower (158 versus 389; P < 0.001). No significant differences were found for SNRb on LGE and both SNRm and CNRm on T1 (P ≥ 0.215). SIQ of O-1T was not significantly different from that of C-1.5T for both R1 and R2 for cine, T1, and LGE (P ≥ 0.157); for T2-STIR, SIQ of O-1T was significantly lower (P = 0.003). R1-R2 concordance was almost perfect (κ = 0.816-0.894), and all examinations were diagnostic. CONCLUSION: Even though quantitative measurements mostly favored C-1.5T, the SIQ of O-1T was not significantly different for any sequence, with the only exception of T2-STIR.

A machine learning approach for differentiating malignant from benign enhancing foci on breast MRI.

BACKGROUND: Differentiate malignant from benign enhancing foci on breast magnetic resonance imaging (MRI) through radiomic signature. METHODS: Forty-five enhancing foci in 45 patients were included in this retrospective study, with needle biopsy or imaging follow-up serving as a reference standard. There were 12 malignant and 33 benign lesions. Eight benign lesions confirmed by over 5-year negative follow-up and 15 malignant histopathologically confirmed lesions were added to the dataset to provide reference cases to the machine learning analysis. All MRI examinations were performed with a 1.5-T scanner. One three-dimensional T1-weighted unenhanced sequence was acquired, followed by four dynamic sequences after intravenous injection of 0.1 mmol/kg of gadobenate dimeglumine. Enhancing foci were segmented by an expert breast radiologist, over 200 radiomic features were extracted, and an evolutionary machine learning method ("training with input selection and testing") was applied. For each classifier, sensitivity, specificity and accuracy were calculated as point estimates and 95% confidence intervals (CIs). RESULTS: A k-nearest neighbour classifier based on 35 selected features was identified as the best performing machine learning approach. Considering both the 45 enhancing foci and the 23 additional cases, this classifier showed a sensitivity of 27/27 (100%, 95% CI 87-100%), a specificity of 37/41 (90%, 95% CI 77-97%), and an accuracy of 64/68 (94%, 95% CI 86-98%). CONCLUSION: This preliminary study showed the feasibility of a radiomic approach for the characterisation of enhancing foci on breast MRI.

Decreased amount of vimentin N-terminal truncated proteolytic products in parkin-mutant skin fibroblasts.

Vimentin, a member of cytoskeleton intermediate filaments proteins, plays a critical role in cell structure and dynamics. The present proteomic study reveals reduced amount of six different lengths, N-terminal truncated proteolytic products of vimentin, in the primary skin fibroblasts from two unrelated PD patients, as compared to control fibroblasts. The decreased amount of N-terminal truncated forms of vimentin in parkin-mutant fibroblasts, could contribute to impairment of cellular function, potentially contributing to the pathogenesis of Parkinson disease.

Allosteric Cooperativity in Proton Energy Conversion in A1-Type Cytochrome c Oxidase.

Cytochrome c oxidase (CcO), the CuA, heme a, heme a3, CuB enzyme of respiratory chain, converts the free energy released by aerobic cytochrome c oxidation into a membrane electrochemical proton gradient (ΔµH+). ΔµH+ derives from the membrane anisotropic arrangement of dioxygen reduction to two water molecules and transmembrane proton pumping from a negative (N) space to a positive (P) space separated by the membrane. Spectroscopic, potentiometric, and X-ray crystallographic analyses characterize allosteric cooperativity of dioxygen binding and reduction with protonmotive conformational states of CcO. These studies show that allosteric cooperativity stabilizes the favorable conformational state for conversion of redox energy into a transmembrane ΔµH+.

Localization accuracy of robotic radiosurgery in 1-view tracking.

PURPOSE: When a lung lesion is detected by only one couple of X-ray tube and image detector integrated with CyberKnife®, the fiducial-less tracking is limited to 1-view (34% of lung treatments at Centro Diagnostico Italiano). The aim of the study was mainly to determine the margin needed to take into account the localization uncertainty along the blind view (out-of-plane direction). METHODS: 36 patients treated in 2-view tracking modality (127 fractions in total) were included in the study. The actual tumor positions were determined retrospectively through logfile analysis and were projected onto 2D image planes. In the same plots the planned target positions based on biphasic breath-hold CT scans were represented preserving the metric with respect to the imaging center. The internal margin necessary to cover in out-of-plane direction the 95% of the target position distribution in the 95% of cases was calculated by home-made software in Matlab®. A validation test was preliminarily performed using XLT Phantom (CIRS) both in 2-view and 1-view scenarios. RESULTS: The validation test proved the reliability of the method, in spite of some intrinsic limitations. Margins were estimated equal to 5 and 6 mm for targets in upper and lower lobe respectively. Biphasic breath-hold CT led to underestimate the target movement in the hypothetical out-of-plane direction. The inter-fractional variability of spine-target distance was an important source of uncertainty for 1-view treatments. CONCLUSION: This graphic comparison method preserving metric could be employed in the clinical workflow of 1-view treatments to get patient-related information for customized margin definition.

Increased Levels of cAMP by the Calcium-Dependent Activation of Soluble Adenylyl Cyclase in Parkin-Mutant Fibroblasts.

Almost half of autosomal recessive early-onset parkinsonism has been associated with mutations in PARK2, coding for parkin, which plays an important role in mitochondria function and calcium homeostasis. Cyclic adenosine monophosphate (cAMP) is a major second messenger regulating mitochondrial metabolism, and it is strictly interlocked with calcium homeostasis. Parkin-mutant (Pt) fibroblasts, exhibiting defective mitochondrial respiratory/OxPhos activity, showed a significant higher value of basal intracellular level of cAMP, as compared with normal fibroblasts (CTRL). Specific pharmacological inhibition/activation of members of the adenylyl cyclase- and of the phosphodiesterase-families, respectively, as well as quantitative reverse transcription polymerase chain reaction (RT-qPCR) analysis, indicate that the higher level of cAMP observed in Pt fibroblasts can contribute to a higher level of activity/expression by soluble adenylyl cyclase (sAC) and to low activity/expression of the phosphodiesterase isoform 4 (PDE4). As Ca2+ regulates sAC, we performed quantitative calcium-fluorimetric analysis, showing a higher level of Ca2+ in the both cytosol and mitochondria of Pt fibroblasts as compared with CTRL. Most notably, inhibition of the mitochondrial Ca2+ uniporter decreased, specifically the cAMP level in PD fibroblasts. All together, these findings support the occurrence of an altered mitochondrial Ca2+-mediated cAMP homeostasis in fibroblasts with the parkin mutation.