The evolution of brain neuron numbers in amniotes

Reconstructing the evolution of brain information-processing capacity is paramount for understanding the rise of complex cognition. Comparative studies of brain evolution typically use brain size as a proxy. However, to get a less biased picture of the evolutionary paths leading to high cognitive power, we need to compare brains not by mass but by numbers of neurons, which are their basic computational units. This study reconstructs the evolution of brains across amniotes by directly analyzing neuron numbers by using the largest dataset of its kind and including essential data on reptiles. We show that reptiles have not only small brains relative to body size but also low neuronal densities, resulting in average neuron numbers over 20 times lower than those in birds and mammals of similar body size. Amniote brain evolution is characterized by the following four major shifts in neuron–brain scaling. The most dramatic increases in brain neurons occurred independently with the appearance of birds and mammals, resulting in convergent neuron scaling in the two endotherm lineages. The other two major increases in the number of neurons happened in core land birds and anthropoid primates, which are two groups known for their cognitive prowess. Interestingly, relative brain size is associated with relative neuronal cell density in reptiles, birds, and primates but not in other mammals. This has important implications for studies using relative brain size as a proxy when looking for evolutionary drivers of animal cognition.

The evolution of cognitive capacity or “intelligence” and its underlying neural substrate has been of long-standing interest to biologists. Great strides have been made in understanding the evolution of brain size in vertebrates, with studies analyzing data on thousands of species (1–3). Since larger animals have larger brains but are not necessarily smarter, most studies of cognitive evolution use relative brain size (corrected for body size), which is thought to reflect extra neurons beyond those needed for controlling the body (4). We now have a good idea where major changes in brain–body scaling happened within birds (2) and mammals (3), and it is also clear that both mammals and birds have relatively larger brains than nonavian sauropsids (hereafter referred to as reptiles), although this has been rarely formally quantified because data on reptilian brain sizes are scarce (5).However, we still lack a clear picture of the evolution of actual brain processing capacity. This is because the same increase in relative brain size can be reached by different evolutionary paths, not always involving actual brain enlargement, and might often result from selection on body size (3). Moreover, similarly sized brains of distantly related species can harbor substantially different numbers of neurons overall and in major brain parts (6, 7). These two caveats invalidate the very idea that we can estimate extra neurons and glean information about cognitive capacity from absolute or relative brain size alone.This capacity is better determined by the number of neurons in the brain or specific brain parts (although their relative importance is still debated), their connections, interneuronal distance, and axonal conduction velocity (8, 9). Unlike brain size, though, these measures are not readily available for a sufficient number of species to be of practical use. Nevertheless, thanks to methodological advances (10), neuronal scaling rules (the allometric relationship between brain mass and neuron numbers) have now been determined for eight high-level mammalian clades (6, 11–13) as well as for a limited sampling of birds (14, 15).To get the big picture of amniote brain evolution, we have to include data on nonavian reptiles. The deepest split in amniote evolution occurred between the synapsid lineage, leading to mammals, and the sauropsid lineage, including reptiles and birds. We cannot tell if similarities between birds and mammals are due to shared ancestry or convergent evolution without considering reptiles. Yet, the dearth of quantitative data on reptile brains is striking—brain mass is available for 183 species (5, 16), compared to thousands for birds and mammals, and neuron numbers are known for a mere 4 reptile species (17–19).Taken together, to understand the evolution of brain processing capacity in amniotes, we need to include nonavian reptiles, consider changes in both brain–body and neuron–brain scaling, and examine the allocation of neurons to different brain parts. In this study, we provide these much needed data and reconstruct the big picture of brain evolution in amniotes in terms of neuron numbers.     

Clinical determinants of primary and secondary fatigue in patients with Parkinson’s disease

Clinical and psychosocial factors associated separately with primary and secondary fatigue in Parkinson’s disease (PD) patients have not been thoroughly studied before. The aim of our study was to assess factors associated with different fatigue domains in groups with primary and secondary fatigue in PD separately. We divided 165 non-demented PD patients according to the absence/presence of depression, anxiety and excessive somnolence into groups with primary fatigue (N = 63) and with secondary fatigue (N = 102). Fatigue domains examined using the multidimensional fatigue inventory were associated through multiple linear regression analyses for each group separately with sociodemographic data, disease duration, functional status as assessed by the Unified Parkinson’s Disease Rating Scale, treatment, depression, anxiety, excessive somnolence and sleep quality. Out of the assessed non-motor symptoms, fatigue was the most frequent (77.6 %). The prevalence of fatigue in the secondary fatigue group was significantly higher than in the primary fatigue group. Both fatigue groups differed significantly in factors associated with different fatigue domains. Functional status or other disease-related factors were not associated with primary fatigue. In the secondary fatigue group, we found associations between some fatigue domains and functional status, older age, male gender and higher anxiety scores. To our knowledge, this is the first study to separately describe clinical determinants and psychosocial factors associated with different fatigue domains in primary and secondary fatigue in PD, underlining the importance of distinguishing primary and secondary fatigue in future PD studies and clinical practice.     

Differential involvement of 5-HT(2A) receptors in the discriminative-stimulus effects of cocaine and methamphetamine

Involvement of 5-HT(2A) receptors in the discriminative-stimulus effects of cocaine versus methamphetamine was studied in Sprague Dawley rats (n=10) trained to discriminate 10 mg/kg cocaine, i.p., from saline under a fixed-ratio 10 (FR10) schedule of food presentation. The ability of (+/-)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI), a 5-HT(2A) receptor agonist, and ketanserin, a 5-HT(2A) receptor antagonist, to either substitute for or block the discriminative-stimulus effects of cocaine, or to shift the cocaine dose-response curve, was evaluated. DOI (0.18-1.0 mg/kg) partially substituted for the training dose of 10 mg/kg cocaine, but only at doses that decreased rates of responding. At the highest dose of DOI tested (1.0 mg/kg), there was about 65% cocaine-appropriate responding. Substitution of DOI for cocaine and DOI-induced decreases in rates of responding were completely reversed by ketanserin (3.0 mg/kg). Ketanserin (3.0 mg/kg) also produced a significant shift to the right of the cocaine dose-response curve and antagonized increases in rates of responding produced by lower doses of cocaine. Ketanserin (1.0-10.0 mg/kg), however, did not block the discriminative-stimulus effects of the training dose of cocaine. When DOI (0.3 mg/kg) was co-administered with different doses of cocaine, there was a slight leftward shift in the cocaine dose-response curve, which was not significant and appeared to reflect simple additive effects of DOI and cocaine. In contrast, the same dose of DOI (0.3 mg/kg) produced a marked and highly significant shift to the left of the methamphetamine (0.18-1.0 mg/kg) dose-response curve in the same subjects and the effects of DOI and methamphetamine were clearly more than additive. The present findings provide new evidence that there is some serotonergic modulation of cocaine's discriminative-stimulus actions, which appears to involve stimulation of 5-HT(2A) receptors. However, involvement of 5-HT(2A) receptor activity in the discriminative-stimulus actions of cocaine appears to be less pronounced than in similar actions of methamphetamine.     

Disturbances of anticoagulation and fibrinolytic systems in monoclonal gammopathies-another mechanism of M-protein interference with hemostasis

Monoclonal gammopathies (MG) may be associated with unique monoclonal immunoglobulin (MIg)-induced disturbances of either primary hemostasis or plasma coagulation. We have investigated the possible interference of MIg with antithrombotic systems in 49 patients with MG. Although an increase of tissue-type plasminogen activator (t-PA) activity was the most frequent abnormality in our group, defect of anticoagulation factors was found in 26.5% of patients. The relationship between MIg type and concentration and frequency of antithrombotic factor abnormalities was not found. The risk of venous thrombosis was higher in patients with the defect in comparison with the unaffected group (46% vs. 22%), but the difference was not statistically significant. Bleeding complications were markedly less frequent in the group of patients with defect of anticoagulation mechanisms (0% vs. 17%). In conclusion, we have found abnormalities in anticoagulation and/or fibrinolytic system, analogous to well-known disturbances of hemostatic mechanisms, in more than a quarter of patients with MG. The interference of M-protein with antithrombotic pathways is supposed to be another mechanism of secondary deficiencies of antithrombin III (AT III), protein C (PC), protein S (PS), plasminogen and APC resistance. Together with other factors, it could contribute to higher risk of thromboembolism in myeloma patients.     

Microsatellite instability in hematological malignancies

The replication error (RER+) phenotype, characterized by microsatellite instability (MSI) has been recently related to mutations of genes involved in DNA mismatch repair pathway. These genetic alterations were first described in hereditary non polyposis colorectal cancer (HNPCC). We examined 44 patients with hematological malignancies (27 AML, 9 MDS, 2 CML-BP and 6 T-ALL) for evidence of MSI. Twenty seven percent of our patients showed differences for only one marker. In four cases (9.1%) MSI was observed in multiple markers and these cases were described as RER+ phenotype. Presented data suggest that this phenomenon may play a role in at least a subset of patients with hematological malignancies.     

Pharmacogenetics of cyclophosphamide in patients with hematological malignancies.

PURPOSE: A high degree of interindividual variation in cyclophosphamide (CPA) pharmacokinetics was reported in certain cancer patient groups. To better understand the mechanisms underlying the variation in CPA metabolism, we have investigated the pharmacokinetics of CPA and its active metabolite 4-hydroxycyclophosphamide (4-OH-CPA) in patients with hematological tumors. The pharmacokinetics of CPA and its active metabolite were related to the genotype of CYP2B6, CYP2C9 and CYP2C19. The influence of liver function on CPA metabolism was also evaluated. METHODS: Twenty-nine patients with hematological malignancies (MM, ALL or NHL) treated with a conventional CPA dose (1g/m(2)) were recruited to this study. Blood samples were collected before, during and after CPA treatment. HPLC was used to measure plasma concentrations of CPA and 4-OH-CPA. Patients were genotyped for the CYP2B6 G516T, CYP2C9*2, CYP2C9*3, CYP2C19*2 and CYP2C19*3 alleles. Serum bilirubin levels were measured before the treatment. Data was analyzed individually and by population pharmacokinetic methods, using non-linear mixed effect modeling. RESULTS: The interindividual variability in exposure to CPA, 4-OHCPA and 4-OH-CPA/CPA was 5.8-, 3.3- and 10.3-fold, respectively. A positive correlation between half-lives of CPA and 4-OH-CPA was found while a significant negative correlation between AUCs of CPA and 4-OH-CPA was detected. In the population analysis, the CYP2B6 G516T variant allele contribution to CPA clearance was about twice as the contribution from the wild type gene while the genotype of CYP2C9 and CYP2C19 did not influence clearance. A negative correlation was observed between bilirubin level and CPA bioactivation. CONCLUSION: This study demonstrates for the first time that the presence of the CYP2B6 G516T mutation increases the rate of 4-OH-CPA formation in patients with hematological malignancies. The liver function prior therapy as assessed by s-bilirubin influences CPA metabolism.