Preclinical and dose-ranging assessment of hESC-derived dopaminergic progenitors for a clinical trial on Parkinson's disease.

Human embryonic stem cell (hESC)-derived midbrain dopaminergic (mDA) cell transplantation is a promising therapeutic strategy for Parkinson's disease (PD). Here, we present the derivation of high-purity mDA progenitors from clinical-grade hESCs on a large scale under rigorous good manufacturing practice (GMP) conditions. We also assessed the toxicity, biodistribution, and tumorigenicity of these cells in immunodeficient rats in good laboratory practice (GLP)-compliant facilities. Various doses of mDA progenitors were transplanted into hemi-parkinsonian rats, and a significant dose-dependent behavioral improvement was observed with a minimal effective dose range of 5,000-10,000 mDA progenitor cells. These results provided insights into determining a low cell dosage (3.15 million cells) for human clinical trials. Based on these results, approval for a phase 1/2a clinical trial for PD cell therapy was obtained from the Ministry of Food and Drug Safety in Korea, and a clinical trial for treating patients with PD has commenced.

Detailed Assessment of 18F-THK5351 Distribution Pattern in the Midbrain: Comparison With Progressive Supranuclear Palsy and Corticobasal Syndrome.

BACKGROUND: 18F-THK5351 PET is used to image ongoing astrogliosis by estimating monoamine oxidase B levels. 18F-THK5351 preferentially accumulates around the substantia nigra (SN) and periaqueductal gray (PG) in the midbrain under healthy conditions and exhibits a "trimodal pattern." In progressive supranuclear palsy (PSP) and corticobasal syndrome (CBS), the midbrain 18F-THK5351 uptake can be increased by astrogliosis, collapsing the "trimodal pattern." We aimed to elucidate cases in which the "trimodal pattern" collapses in PSP and CBS. PATIENTS AND METHODS: Participants in the PSP (n = 11), CBS (n = 17), Alzheimer disease (n = 11), and healthy control (n = 8) groups underwent 18F-THK5351 PET. Volumes of interest (VOIs) were placed on the SN, PG, and their midpoints. The midbrain uptake ratio (MUR) was calculated to assess the trimodal pattern as follows: MUR = (VOI value on the midpoint)/(VOI value on the SN and PG). Approximately, the trimodal pattern can be identified at MUR <1 but not at MUR >1. RESULTS: Compared with the healthy control group, MUR significantly increased in the PSP (P < 0.01) and CBS (P < 0.01) groups, but was unchanged in the Alzheimer disease group (P = 0.10). In the PSP group, all patients, including 2 with mild symptoms and a short disease duration, showed MUR >1. In the CBS group, MUR varied widely. CONCLUSIONS: In PSP, the trimodal pattern can collapse even in the early phase when symptoms are mild. In CBS, the trimodal pattern may or may not collapse depending on the underlying pathology.

Rehemorrhage of brainstem cavernous malformations: a benchmark approach to individualized risk and severity assessment.

OBJECTIVE: Brainstem cavernous malformations (BSCMs) represent a unique subgroup of cavernous malformations with more hemorrhagic presentation and technical challenges. This study aimed to provide individualized assessment of the rehemorrhage clustering risk of BSCMs after the first symptomatic hemorrhage and to identify patients at higher risk of neurological deterioration after new hemorrhage, which would help in clinical decision-making. METHODS: A total of 123 consecutive BSCM patients with symptomatic hemorrhage were identified between 2015 and 2022, with untreated follow-up > 12 months or subsequent hemorrhage during the untreated follow-up. Nomograms were proposed to individualize the assessment of subsequent hemorrhage risk and neurological status (determined by the modified Rankin Scale [mRS] score) after future hemorrhage. The least absolute shrinkage and selector operation (LASSO) regression was used for feature screening. The calibration curve and concordance index (C-index) were used to assess the internal calibration and discrimination performance of the nomograms. Cross-validation was further performed to validate the accuracy of the nomograms. RESULTS: Prior hemorrhage times (adjusted OR [aOR] 6.78 per ictus increase) and Zabramski type I or V (OR 11.04) were associated with rehemorrhage within 1 year. A lower mRS score after previous hemorrhage (aOR 0.38 for a shift to a higher mRS score), Zabramski type I or V (OR 3.41), medulla or midbrain location (aOR 2.77), and multiple cerebral cavernous malformations (aOR 11.76) were associated with worsened neurological status at subsequent hemorrhage. The nomograms showed good accuracy and discrimination, with a C-index of 0.80 for predicting subsequent hemorrhage within 1 year and 0.71 for predicting neurological status after subsequent hemorrhage, which were maintained in cross-validation. CONCLUSIONS: An individualized approach to risk and severity assessment of BSCM rehemorrhage was feasible with clinical and imaging features.

A novel two-factor monosynaptic TRIO tracing method for assessment of circuit integration of hESC-derived dopamine transplants.

Transplantation in Parkinson's disease using human embryonic stem cell (hESC)-derived dopaminergic (DA) neurons is a promising future treatment option. However, many of the mechanisms that govern their differentiation, maturation, and integration into the host circuitry remain elusive. Here, we engrafted hESCs differentiated toward a ventral midbrain DA phenotype into the midbrain of a preclinical rodent model of Parkinson's disease. We then injected a novel DA-neurotropic retrograde MNM008 adeno-associated virus vector capsid, into specific DA target regions to generate starter cells based on their axonal projections. Using monosynaptic rabies-based tracing, we demonstrated for the first time that grafted hESC-derived DA neurons receive distinctly different afferent inputs depending on their projections. The similarities to the host DA system suggest a previously unknown directed circuit integration. By evaluating the differential host-to-graft connectivity based on projection patterns, this novel approach offers a tool to answer outstanding questions regarding the integration of grafted hESC-derived DA neurons.

Inactivation mode of sodium channels defines the different maximal firing rates of conventional versus atypical midbrain dopamine neurons.

Two subpopulations of midbrain dopamine (DA) neurons are known to have different dynamic firing ranges in vitro that correspond to distinct projection targets: the originally identified conventional DA neurons project to the dorsal striatum and the lateral shell of the nucleus accumbens, whereas an atypical DA population with higher maximum firing frequencies projects to prefrontal regions and other limbic regions including the medial shell of nucleus accumbens. Using a computational model, we show that previously identified differences in biophysical properties do not fully account for the larger dynamic range of the atypical population and predict that the major difference is that originally identified conventional cells have larger occupancy of voltage-gated sodium channels in a long-term inactivated state that recovers slowly; stronger sodium and potassium conductances during action potential firing are also predicted for the conventional compared to the atypical DA population. These differences in sodium channel gating imply that longer intervals between spikes are required in the conventional population for full recovery from long-term inactivation induced by the preceding spike, hence the lower maximum frequency. These same differences can also change the bifurcation structure to account for distinct modes of entry into depolarization block: abrupt versus gradual. The model predicted that in cells that have entered depolarization block, it is much more likely that an additional depolarization can evoke an action potential in conventional DA population. New experiments comparing lateral to medial shell projecting neurons confirmed this model prediction, with implications for differential synaptic integration in the two populations.

The cost of obtaining rewards enhances the reward prediction error signal of midbrain dopamine neurons.

Midbrain dopamine neurons are known to encode reward prediction errors (RPE) used to update value predictions. Here, we examine whether RPE signals coded by midbrain dopamine neurons are modulated by the cost paid to obtain rewards, by recording from dopamine neurons in awake behaving monkeys during performance of an effortful saccade task. Dopamine neuron responses to cues predicting reward and to the delivery of rewards were increased after the performance of a costly action compared to a less costly action, suggesting that RPEs are enhanced following the performance of a costly action. At the behavioral level, stimulus-reward associations are learned faster after performing a costly action compared to a less costly action. Thus, information about action cost is processed in the dopamine reward system in a manner that amplifies the following dopamine RPE signal, which in turn promotes more rapid learning under situations of high cost.

Expression and function of microRNA-9 in the mid-hindbrain area of embryonic chick.

BACKGROUND: MiR-9 is a small non-coding RNA that is highly conserved between species and primarily expressed in the central nervous system (CNS). It is known to influence proliferation and neuronal differentiation in the brain and spinal cord of different vertebrates. Different studies have pointed to regional and species-specific differences in the response of neural progenitors to miR-9. METHODS: In ovo and ex ovo electroporation was used to overexpress or reduce miR-9 followed by mRNA in situ hybridisation and immunofluorescent stainings to evaluate miR- expression and the effect of changed miR-9 expression. RESULTS: We have investigated the expression and function of miR-9 during early development of the mid-hindbrain region (MH) in chick. Our analysis reveals a closer relationship of chick miR-9 to mammalian miR-9 than to fish and a dynamic expression pattern in the chick neural tube. Early in development, miR-9 is diffusely expressed in the entire brain, bar the forebrain, and it becomes more restricted to specific areas of the CNS at later stages. MiR-9 overexpression at HH9-10 results in a reduction of FGF8 expression and premature neuronal differentiation in the mid-hindbrain boundary (MHB). Within the midbrain miR-9 does not cause premature neuronal differentiation it rather reduces proliferation in the midbrain. CONCLUSION: Our findings indicate that miR-9 has regional specific effects in the developing mid-hindbrain region with a divergence of response of regional progenitors.

Assessment of Ketamine Binding of the Serotonin Transporter in Humans with Positron Emission Tomography.

Background: Comprehensive description of ketamine's molecular binding profile becomes increasingly pressing as use in real-life patient cohorts widens. Animal studies attribute a significant role in the substance's antidepressant effects to the serotonergic system. The serotonin transporter is a highly relevant target in this context, because it is central to depressive pathophysiology and treatment. This is, to our knowledge, the first study investigating ketamine's serotonin transporter binding in vivo in humans. Methods: Twelve healthy subjects were assessed twice using [11C]DASB positron emission tomography. A total of 0.50 mg/kg bodyweight ketamine was administered once i.v. prior to the second positron emission tomography scan. Ketamine plasma levels were determined during positron emission tomography. Serotonin transporter nondisplaceable binding potential was computed using a reference region model, and occupancy was calculated for 4 serotonin transporter-rich regions (caudate, putamen, thalamus, midbrain) and a whole-brain region of interest. Results: After administration of the routine antidepressant dose, ketamine showed <10% occupancy of the serotonin transporter, which is within the test-retest variability of [11C]DASB. A positive correlation between ketamine plasma levels and occupancy was shown. Conclusions: Measurable occupancy of the serotonin transporter was not detectable after administration of an antidepressant dose of ketamine. This might suggest that ketamine binding of the serotonin transporter is unlikely to be a primary antidepressant mechanism at routine antidepressant doses, as substances that facilitate antidepressant effects via serotonin transporter binding (e.g., selective serotonin reuptake inhibitors) show 70% to 80% occupancy. Administration of high-dose ketamine is widening. Based on the positive relationship we find between ketamine plasma levels and occupancy, there is a need for investigation of ketamine's serotonin transporter binding at higher doses.

Lower neighborhood quality in adolescence predicts higher mesolimbic sensitivity to reward anticipation in adulthood.

Life history theory suggests that adult reward sensitivity should be best explained by childhood, but not current, socioeconomic conditions. In this functional magnetic resonance imaging (fMRI) study, 83 participants from a larger longitudinal sample completed the monetary incentive delay (MID) task in adulthood (∼25 years old). Parent-reports of neighborhood quality and parental SES were collected when participants were 13 years of age. Current income level was collected concurrently with scanning. Lower adolescent neighborhood quality, but neither lower current income nor parental SES, was associated with heightened sensitivity to the anticipation of monetary gain in putative mesolimbic reward areas. Lower adolescent neighborhood quality was also associated with heightened sensitivity to the anticipation of monetary loss activation in visuo-motor areas. Lower current income was associated with heightened sensitivity to anticipated loss in occipital areas and the operculum. We tested whether externalizing behaviors in childhood or adulthood could better account for neighborhood quality findings, but they did not. Findings suggest that neighborhood ecology in adolescence is associated with greater neural reward sensitivity in adulthood above the influence of parental SES or current income and not mediated through impulsivity and externalizing behaviors.

HEPeak: an HMM-based exome peak-finding package for RNA epigenome sequencing data.

BACKGROUND: Methylated RNA Immunoprecipatation combined with RNA sequencing (MeRIP-seq) is revolutionizing the de novo study of RNA epigenomics at a higher resolution. However, this new technology poses unique bioinformatics problems that call for novel and sophisticated statistical computational solutions, aiming at identifying and characterizing transcriptome-wide methyltranscriptome. RESULTS: We developed HEP, a Hidden Markov Model (HMM)-based Exome Peak-finding algorithm for predicting transcriptome methylation sites using MeRIP-seq data. In contrast to exomePeak, our previously developed MeRIP-seq peak calling algorithm, HEPeak models the correlation between continuous bins in an m6A peak region and it is a model-based approach, which admits rigorous statistical inference. HEPeak was evaluated on a simulated MeRIP-seq dataset and achieved higher sensitivity and specificity than exomePeak. HEPeak was also applied to real MeRIP-seq datasets from human HEK293T cell line and mouse midbrain cells and was shown to be able to recapitulate known m6A distribution in transcripts and identify novel m6A sites in long non-coding RNAs. CONCLUSIONS: In this paper, a novel HMM-based peak calling algorithm, HEPeak, was developed for peak calling for MeRIP-seq data. HEPeak is written in R and is publicly available.

Assessment of fetal midbrain and hindbrain in mid-sagittal cranial plane by three-dimensional multiplanar sonography. Part 2: application of nomograms to fetuses with posterior fossa malformations.

OBJECTIVES: To apply fetal midbrain (MB) and hindbrain (HB) nomograms, developed using three-dimensional multiplanar sonographic reconstruction (3D-MPR) in the mid-sagittal cranial plane, to fetuses with known posterior fossa malformations. METHODS: In this retrospective study we examined sonographic volumes obtained by sagittal acquisition in 43 fetuses diagnosed with posterior fossa abnormalities and evaluated in the mid-sagittal cranial plane, using 3D-MPR, the following: MB parameters tectal length (TL) and anteroposterior midbrain diameter (APMD), and HB parameters anteroposterior pons diameter (APPD), superoinferior vermian diameter (SIVD) and anteroposterior vermian diameter (APVD). Fetuses were grouped, according to malformation, into eight categories: cobblestone malformation complex (CMC, n = 3), Chiari-II malformation (C-II, n = 7), pontocerebellar hypoplasia (PCH, n = 2), rhombencephalosynapsis (RES, n = 4), Dandy-Walker malformation (n = 8), vermian dysgenesis (VD, n = 7), persistent Blake's pouch cyst (n = 6) and megacisterna magna (n = 6). In each case and for each subgroup, the MB-HB biometric parameters and their z-scores were evaluated with reference to our new nomograms. RESULTS: The new MB-HB nomograms were able to identify the brainstem and vermian anomalies and differentiate fetuses with MB-HB malformations from those with isolated enlarged posterior fossa cerebrospinal fluid spaces. Use of the nomograms enabled detection of an elongated tectum in fetuses with CMC, C-II and RES, and a flattened pontine belly in cases of CMC, PCH and VD. In the fetuses with VD, the nomograms enabled division into three distinctive groups: (1) those with small SIVD and APVD, (2) those with normal SIVD but small APVD, and (3) those with small SIVD but normal APVD. CONCLUSIONS: Application of our new reference data, that for the first time include the MB, enables accurate diagnosis of brain malformations affecting the MB and HB and makes possible novel characterization of previously described features of posterior fossa anomalies.

Assessment of fetal midbrain and hindbrain in mid-sagittal cranial plane by three-dimensional multiplanar sonography. Part 1: comparison of new and established nomograms.

OBJECTIVE: To construct nomograms for fetal midbrain (MB) and hindbrain (HB) dimensions, assessed in the mid-sagittal cranial plane by three-dimensional multiplanar sonographic reconstruction (3D-MPR). METHODS: This was a prospective cross-sectional study of 334 healthy fetuses in low-risk singleton pregnancies between 16 and 35 gestational weeks. All sonographic volumes were obtained by sagittal acquisition. The following MB and HB parameters were evaluated in the mid-sagittal cranial plane using 3D-MPR: MB parameters tectal length (TL) and anteroposterior midbrain diameter (APMD), and HB parameters anteroposterior pons diameter (APPD), superoinferior vermian diameter (SIVD), anteroposterior vermian diameter (APVD) and anteroposterior diameter of the fourth ventricle (APDFV). The measurements were presented as growth charts according to gestational age. RESULTS: MB and HB biometry were best assessed between 19 and 29 weeks. During this period, adequate visualization was achieved for successful measurement of TL in 90.9% of cases, APMD in 86.6%, APPD in 73.7%, SIVD in 74.2%, APVD in 71% and APDFV in 71%. There was a linear growth pattern, with Pearson correlation coefficients of 0.79 for TL, 0.88 for APMD, 0.91 for APPD, 0.95 for SIVD, 0.88 for APVD and 0.88 for APDFV (P < 0.0001 for each). The mean intra- and interobserver variations for the MB measurements and vermian diameters ranged between 4.3% and 9%. APPD and APDFV showed highest mean variations: 9.0% and 19.4% (intraobserver) and 11.6% and 17.7% (interobserver), respectively. CONCLUSION: We present new nomograms for assessment of the fetal MB and HB using 3D-MPR in the mid-sagittal cranial plane. To our knowledge, these are the first proposed nomograms for fetal MB dimensions.

Quantitative assessment of iron deposition in the midbrain using 3D-enhanced T2 star weighted angiography (ESWAN): a preliminary cross-sectional study of 20 Parkinson's disease patients.

Conventional magnetic resonance imaging (MRI) assesses neurodegenerative structural changes in the cerebral anatomy of Parkinson's disease (PD) patients but cannot detect non-structural abnormalities; however, enhanced T2 star weighted angiography (ESWAN) can precisely indicate PD-related substantia nigra (SN) iron deposition. The differences in ESWAN-based parameters between different PD stages were assessed using midbrain iron deposits of 20 PD patients aged 64.3±12.7 (41-85) years grouped by Hoehn and Yahr staging into minimal (stages ≤2.5) or moderate to severe (stages ≥3.0) motor impairment groups and 14 healthy control subjects. Conventional MRI and ESWAN measurements of mean phase value (MPV) and midbrain dimensions (width and diameter) revealed similar anatomical characteristics; however, ESWAN revealed the presence of smaller MPVs and SN pars compacta (SNc) (P<0.01) and a negative correlation between reduction extent and motor impairment (P<0.01). SNc width to midbrain diameter was reduced in moderate to severe impairment patients versus control and minimal impairment patients (both P<0.01). A positive correlation was found between MPV and width or SNc width to midbrain diameter ratio (P<0.01 and P<0.05, respectively). Minimal impairment group mean MPV and substantia nigra pars reticulata (SNr) width evidenced no significant reduction, unlike significant reductions in the moderate to severe impairment group (P<0.01). No significant changes were observed in MPV or width in the RN region (P>0.05). ESWAN allows for early and accurate iron deposition determination in PD patients, particularly useful as a supplement to conventional MRI in early-stage PD patients.

Limited encoding of effort by dopamine neurons in a cost-benefit trade-off task.

Animals are thought to evaluate the desirability of action options using a unified scale that combines predicted benefits ("rewards"), costs, and the animal's internal motivational state. Midbrain dopamine neurons have long been associated with the reward part of this equation, but it is unclear whether these neurons also estimate the costs of taking an action. We studied the spiking activity of dopamine neurons in the substantia nigra pars compacta of monkeys (Macaca mulatta) during a reaching task in which the energetic costs incurred (friction loads) and the benefits gained (drops of food) were manipulated independently. Although the majority of dopamine neurons encoded the upcoming reward alone, a subset predicted net utility of a course of action by signaling the expected reward magnitude discounted by the invested cost in terms of physical effort. In addition, the tonic activity of some dopamine neurons was slowly reduced in conjunction with the accumulated trials, which is consistent with the hypothesized role for tonic dopamine in the invigoration or motivation of instrumental responding. The present results shed light on an often-hypothesized role for dopamine in the regulation of the balance in natural behaviors between the energy expended and the benefits gained, which could explain why dopamine disorders, such as Parkinson's disease, lead to a breakdown of that balance.

The imaging diagnosis and prognosis assessment of patients with midbrain injury in the acute phase of craniocerebral injury.

We prospectively studied the difference between head CT and MRI in the detection of midbrain injury at the acute stage, the characteristics of MRS in the midbrain, and its relationship to the prognosis. The aim of this study is to propose the imaging diagnosis and outcome assessment indicators for midbrain injury.According to the clinical diagnosis standard, 22 patients with midbrain injury were chosen as a midbrain injury group,and 20 cases with craniocerebral injury without brain stem injury as the control group,10 normal adult volunteers as the normal control group. CT was performed on days 1, 3, 5, and 7 respectively,and MRI and MRS within 7 days post-injury. All patients were followed up for 6 months post-injury.The positive diagnosis rate of 63.64% in MRI for midbrain injury was significantly higher than that of 13.63% found in CT. MRI showed that the location of the midbrain injury was closely associated with prognosis. The reduction of NAA/Cr or NAA/Cho ratio was more obvious and the prognosis of the patients poorer. Midbrain injury can be diagnosed more clearly and its severity or prognosis could also be evaluated by MRI and MRS.

Cine-PC MR in assessment of cerebrospinal fluid velocity in the aqueduct of the midbrain correlated with intracranial pressure--initial study.

We assessed the changes in cerebrospinal fluid (CSF) hydrodynamics caused by barriers to CSF circulation, and determined the relationship between CSF velocity and intracranial pressure in the aqueduct of the midbrain. This was determined by correlating the CSF peak flow velocity with the intracranial pressure (ICP) obtained from a lumbar puncture (LP) procedure. The CSF peak flow velocity was measured by finger pulse-gated cine-phase contrast (PC) MR scan 8-12 hours after LP was performed in 28 patients. All patients were divided into 2 groups based on the directional patterns of the CSF net flow in the aqueduct of the midbrain over one cardiac cycle. The CSF peak net velocity (V(net)) was then correlated with ICP utilizing Pearson correlation analysis method, with significance difference assigned at the 5% level. Routine MR scanning revealed no abnormal findings in the brain when the direction of the CSF net flow is caudal. V(net) in the aqueduct of the midbrain was correlated positively with ICP (y(V)=0.011+0.002 ×(ICP), r=0.69, p<0.01). However, varying degrees of the hydrocephalus were observed in those patients who demonstrated a cranial direction of the CSF net flow. Our results indicate that non-invasive measurement of the CSF peak flow with cine-PC MR imaging can be related to the change of CSF circulation caused by the obstructions to the CSF circulation in the patients with various neurological disorders. This unique method may be a substantially useful tool to assess the changes in the ICP in the directional pattern.

All four zebrafish Wnt7 genes are expressed during early brain development.

Wnt-signalling is involved in a number of biological processes in the course of embryonic development, cell fate determination, proliferation, stem cell maintenance and oncogenesis. Wnt ligands are secreted glycoproteins and the number of Wnt isoforms varies between five in nematodes and 27 in fish. The highly conserved group of Wnt7 genes has been found to signal via at least three Wnt-signalling pathways dependent on the developmental context. These ligands have been identified as important regulators in a number of processes ranging from formation of bones, lungs, kidneys, reproductive organs and placenta to vasculogenesis and synaptogenesis in the brain. The importance of Wnt7 function is underscored by their implication in disease syndromes in man. Unlike the single Wnt7a and Wnt7b mammalian genes we find that the zebrafish genome contains two paralogues genes for each Wnt7 ligand. Here, we compare these four Wnt7 genes evolutionarily and analyse their expression during the first two days of embryonic development. We find Wnt7 genes mainly expressed in a number of CNS structures at developmental stages at which patterning and neural specification takes place. The timely and spatially overlapping as well as complementary gene expression suggests diverse as well as redundant involvements during brain development.

[Quantitative evaluation of SPECT with FP-CIT. Importance of the reference area]. / Valoración cuantitativa del SPECT con FP-CIT. Importancia del área de referencia.

OBJECTIVE: The aim of this study is to compare several uptake indexes between specific and non-specific activity to determine the existence of degenerative Parkinsonism according to different reference areas. MATERIAL AND METHODS: A total of 46 patients (23 men and 23 women), randomly selected from individuals referred to our center with a movement disorder, were included in the study. Mean age was 70.2 ± 10.2 years (41-87). The uptake indexes were obtained through the areas of interest (ROIs) located in the striate (specific uptake) and other reference ROIs located in areas with different concentrations of serotonin receptors: low-cerebellum, medium-occipital cortex and high-midbrain. RESULTS: A high linear correlation was found between indexes having low and medium concentration of serotonin receptors. The ROC curve analysis shows an area under the curve of 0.874, 0.886 and 0.739 and regression coefficients of 5.41, 6.62 and 3.41, respectively for the striatum/cerebellum (E/C), striatum/occipital (E/O) and striatum midbrain (E/M) indexes. Optimal cutoff for E/O (1.35), index with the best behavior, provides a sensitivity of 0.84 and specificity of 0.89. CONCLUSION: The reference area selected may alter the predictive power of the different indexes to determine the existence of a degenerative Parkinsonism.

Hypothesis testing for neural cell growth experiments using a hybrid branching process model.

Neuron branching patterns can characterize neural cell types and act as markers for neurodegenerative disease and neural development. We develop a hybrid Markovian model for neural branching that extends previously published models by (i) using a discretized gamma model to account for underdispersion in primary branching, (ii) incorporating both bifurcation and trifurcation branching events to accommodate observed data, and (iii) only requiring branch counts and not branching topology as observations, allowing larger numbers of neurons to be sampled than in previous literature. Inference for primary branching is achieved through a gamma generalized linear model. Due to incomplete data, bifurcation and trifurcation probabilities are estimated using an expectation-maximization algorithm, which is shown to give consistent estimates using simulation studies and theoretical arguments. In simulation studies, comparison of standard errors shows no significant loss of accuracy relative to when topological information is available. A unified methodology for testing hypotheses using likelihood ratio tests (LRTs) is developed. The methodology is applied to an experiment where neurons are cocultured with different treatments: growth factor (GF), hypothalamic-astroglial conditioned medium (HY), and combination. The model provides statistically adequate fit at all branching orders. All treatments cause significantly higher branching at primary and secondary orders relative to control (p-value < 0.01), but not at higher branching orders, suggesting genetic regulation by the treatments. Using a computationally feasible lower bound on the LRT, bifurcation probabilities are shown to decrease exponentially with branching order for all treatments except HY (p-value 0.03).