Altered trajectories of neurodevelopment and behavior in mouse models of Rett syndrome.

Rett Syndrome (RTT) is a genetic disorder that is caused by mutations in the x-linked gene coding for methyl-CpG-biding-protein 2 (MECP2) and that mainly affects females. Male and female transgenic mouse models of RTT have been studied extensively, and we have learned a great deal regarding RTT neuropathology and how MeCP2 deficiency may be influencing brain function and maturation. In this manuscript we review what is known concerning structural and coinciding functional and behavioral deficits in RTT and in mouse models of MeCP2 deficiency. We also introduce our own corroborating data regarding behavioral phenotype and morphological alterations in volume of the cortex and striatum and the density of neurons, aberrations in experience-dependent plasticity within the barrel cortex and the impact of MeCP2 loss on glial structure. We conclude that regional structural changes in genetic models of RTT show great similarity to the alterations in brain structure of patients with RTT. These region-specific modifications often coincide with phenotype onset and contribute to larger issues of circuit connectivity, progression, and severity. Although the alterations seen in mouse models of RTT appear to be primarily due to cell-autonomous effects, there are also non-cell autonomous mechanisms including those caused by MeCP2-deficient glia that negatively impact healthy neuronal function. Collectively, this body of work has provided a solid foundation on which to continue to build our understanding of the role of MeCP2 on neuronal and glial structure and function, its greater impact on neural development, and potential new therapeutic avenues.

Oligomerization of Mannan-binding Lectin Dictates Binding Properties and Complement Activation.

The complement system is a part of the innate immune system and is involved in recognition and clearance of pathogens and altered-self structures. The lectin pathway of the complement system is initiated when soluble pattern recognition molecules (PRMs) with collagen-like regions bind to foreign or altered self-surfaces. Associated with the collagen-like stems of these PRMs are three mannan-binding lectin (MBL)-associated serine proteases (MASPs) and two MBL-associated proteins (MAps). The most studied of the PRMs, MBL, is present in serum mainly as trimeric and tetrameric oligomers of the structural subunit. We hypothesized that oligomerization of MBL may influence both the potential to bind to micro organisms and the interaction with the MASPs and MAps, thus influencing the ability to initiate complement activation. When testing binding at 37 °C, we found higher binding of tetrameric MBL to Staphylococcus aureus (S. aureus) than trimeric and dimeric MBL. In serum, we found that tetrameric MBL was the main oligomeric form present in complexes with the MASPs and MAp44. Such preference was confirmed using purified forms of recombinant MBL (rMBL) oligomers, where tetrameric rMBL interacted stronger with all of the MASPs and MAp44, compared to trimeric MBL. As a direct consequence of the weaker interaction with the MASPs, we found that trimeric rMBL was inferior to tetrameric rMBL in activating the complement system. Our data suggest that the oligomeric state of MBL is crucial both for the binding properties and the effector function of MBL.

Inositol polyphosphate multikinase is a transcriptional coactivator required for immediate early gene induction.

Profound induction of immediate early genes (IEGs) by neural activation is a critical determinant for plasticity in the brain, but intervening molecular signals are not well characterized. We demonstrate that inositol polyphosphate multikinase (IPMK) acts noncatalytically as a transcriptional coactivator to mediate induction of numerous IEGs. IEG induction by electroconvulsive stimulation is virtually abolished in the brains of IPMK-deleted mice, which also display deficits in spatial memory. Neural activity stimulates binding of IPMK to the histone acetyltransferase CBP and enhances its recruitment to IEG promoters. Interestingly, IPMK regulation of CBP recruitment and IEG induction does not require its catalytic activities. Dominant-negative constructs, which prevent IPMK-CBP binding, substantially decrease IEG induction. As IPMK is ubiquitously expressed, its epigenetic regulation of IEGs may influence diverse nonneural and neural biologic processes.

The brain-derived neurotrophic factor receptor TrkB is critical for the acquisition but not expression of conditioned incentive value.

Stimuli paired with reward acquire incentive properties that are important for many aspects of motivated behavior, such as feeding and drug-seeking. Here we used a novel chemical-genetic strategy to determine the role of the brain-derived neurotrophic factor (BDNF) receptor TrkB, known to be critical to many aspects of neural development and plasticity, during acquisition and expression of positive incentive value by a cue paired with food. We assessed that cue's learned incentive value in a conditioned reinforcement task, in which its ability to reinforce instrumental responding later, in the absence of food itself, was examined. In TrkB (F616A) knock-in mice, TrkB kinase activity was suppressed by administering the TrkB inhibitor 1NMPP1 during the period of initial cue incentive learning only (i.e. Pavlovian training), during nose-poke conditioned reinforcement testing only, during both phases, or during neither phase. All mice acquired cue-food associations as indexed by approach responses. However, TrkB (F616A) mice that received 1NMPP1 during initial cue incentive learning failed to show conditioned reinforcement of nose-poking, regardless of their treatment in testing, whereas administration of 1NMMP1 only during the testing phase had no effect. The effects of 1NMPP1 administration were due to inhibition of TrkB(F616A), because the performance of wild-type mice was unaffected by administration of the compound during either phase. These data indicate that BDNF or NT4 signaling through TrkB receptors is required for the acquisition of positive incentive value, but is not needed for the expression of previously acquired incentive value in the reinforcement of instrumental behavior.

Genetic background differences and nonassociative effects in mouse trace fear conditioning.

Fear conditioning, including variants such as delay and trace conditioning that depend on different neural systems, is widely used to behaviorally characterize genetically altered mice. We present data from three strains of mice, C57/BL6 (C57), 129/SvlmJ (129), and a hybrid strain of the two (F(1) hybrids), trained on various versions of a trace fear-conditioning protocol. The initial version was taken from the literature but included unpaired control groups to assess nonassociative effects on test performance. We observed high levels of nonassociative freezing in both contextual and cued test conditions. In particular, nonassociative freezing in unpaired control groups was equivalent to freezing shown by paired groups in the tests for trace conditioning. A number of pilot studies resulted in a new protocol that yielded strong context conditioning and low levels of nonassociative freezing in all mouse strains. During the trace-CS test in this protocol, freezing in unpaired controls remained low in all strains, and both the C57s and F(1) hybrids showed reliable associative trace fear conditioning. Trace conditioning, however, was not obtained in the 129 mice. Our findings indicate that caution is warranted in interpreting mouse fear-conditioning studies that lack control conditions to address nonassociative effects. They also reveal a final set of parameters that are important for minimizing such nonassociative effects and demonstrate strain differences across performance in mouse contextual and trace fear conditioning.

Foreign Body Ingestion Causing Recurrent Diverticulitis.

BACKGROUND Ingested foreign bodies (IFBs) are usually asymptomatic and are excreted uneventfully. IFBs become a major concern in elderly patients due to the increase number of diverticuloses where the foreign body can lodge and cause severe complications. CASE REPORT We report a case of an elderly patient who ingested a chicken bone that caused recurrent diverticulitis. CONCLUSIONS The diagnosis of complicated IFB cases is challenging, requires physician clinical expertise, and must be considered in individuals at risk.

Impaired hippocampal-dependent memory and reduced parvalbumin-positive interneurons in a ketamine mouse model of schizophrenia.

The hippocampus of patients with schizophrenia displays aberrant excess neuronal activity which affects cognitive function. Animal models of the illness have recapitulated the overactivity in the hippocampus, with a corresponding regionally localized reduction of inhibitory interneurons, consistent with that observed in patients. To better understand whether cognitive function is similarly affected in these models of hippocampal overactivity, we tested a ketamine mouse model of schizophrenia for cognitive performance in hippocampal- and medial prefrontal cortex (mPFC)-dependent tasks. We found that adult mice exposed to ketamine during adolescence were impaired on a trace fear conditioning protocol that relies on the integrity of the hippocampus. Conversely, the performance of the mice was normal on a delayed response task that is sensitive to mPFC damage. We confirmed that ketamine-exposed mice had reduced parvalbumin-positive interneurons in the hippocampus, specifically in the CA1, but not in the mPFC in keeping with the behavioral findings. These results strengthened the utility of the ketamine model for preclinical investigations of hippocampal overactivity in schizophrenia.

Dimensional assessment of behavioral changes in the cuprizone short-term exposure model for psychosis.

Recent clinical studies have suggested a role for immune/inflammatory responses in the pathophysiology of psychosis. However, a mechanistic understanding of this process and its application for drug discovery is underdeveloped. Here we assessed our recently developed cuprizone short-term exposure (CSE) mouse model across behavioral domains targeting neurocognitive and neuroaffective systems. We propose that the CSE model may be useful for understanding the mechanism associating inflammation and psychosis, with applications for drug discovery in that context.

Cytological behaviour of floral organs and in silico characterization of differentially expressed transcript-derived fragments associated with 'floral bud distortion' in soybean.

An attempt was made to understand the 'floral bud distortion' (FBD), an unexplored disorder prevailing in soybean. Cytological behaviour of floral reproductive organs and in silico characterization of differentially expressed transcript-derived fragments (TDFs) in symptomatic and asymptomatic soybean plants were carried out. Pollens in asymptomatic plants do not have defects in number, size, shape and function. However, in symptomatic plant, pollens were found nonviable, abnormal in shape and with reduced germination ability. Here, we employed a computational approach, exploring invaluable resources. The tissue-specific transcript profile of symptomatic and asymptomatic sources was compared to determine differentially expressed TDFs associated with FBD to improve its basic understanding. A total of 60 decamer primers produced 197 scorable amplicons, ranged 162-1130 bp, of which 171 were monomorphic and 26 were differentially regulated. Reproducible TDFs were sequenced and characterized for their homology analysis, annotation, protein-protein interaction, subcellular localization and their physical mapping. Homology-based annotation of TDFs in soybean revealed presence of two characterized and seven uncharacterized hits. Annotation of characterized sequences showed presence of genes, namely auxin response factor 9 (ARF9) and forkhead-associated (FHA) domain, which are directly involved in plant development through various pathways, such as hormonal regulation, plant morphology, embryogenesis and DNA repair.

Synthesis, spectral characterization, thermal behaviour, antibacterial activity and DFT calculation on N'-[bis(methylsulfanyl) methylene]-2-hydroxybenzohydrazide and N'-(4-methoxy benzoyl)-hydrazinecarbodithioic acid ethyl ester.

Two new compounds N'-[bis(methylsulfanyl) methylene]-2-hydroxybenzohydrazide {Hbmshb (1)} and N'-(4-methoxy benzoyl)-hydrazinecarbodithioic acid ethyl ester {H2mbhce (2)} have been synthesized and characterized with the aid of elemental analyses, IR, NMR and single crystal X-ray diffraction data. Compounds 1 and 2 crystallize in orthorhombic and monoclinic systems with space group Pna21 and P21/n, respectively. Inter and intra molecular hydrogen bonding link two molecules and provide linear chain structure. In addition to this, compound 2 is stabilized by CH⋯π and NH⋯π interactions. Molecular geometry from X-ray analysis, geometry optimization, charge distribution, bond analysis, frontier molecular orbital (FMO) analysis and non-linear optical (NLO) effects have been performed using the density functional theory (DFT) with the B3LYP functional. The bioefficacy of compounds has been examined against the growth of bacteria to evaluate their anti-microbial potential. Compounds 1 and 2 are thermally stable and show NLO behaviour better than the urea crystal.

Transcriptional mechanisms of hippocampal aging.

Aging related cognitive decline is an increasing health problem but affects only a subset of elderly humans. This research uses outbred young (Y) and aged rats. Behavioral characterization distinguishes aged rats with impaired spatial learning (AI) and aged rats with unimpaired learning ability (AU), mimicking the varied susceptibility of the human population to age-associated learning impairment. Studies are testing a hypothesis that hippocampal transcriptional mechanisms and gene expression profiles linked to activator protein-1 (AP-1) and glucocorticoid receptor (GR), mineralocorticoid receptor (MR) or cyclic AMP response element binding protein (CREB) families of transcription factors distinguish successful or unsuccessful aging and cognition. Results from mRNA assays, in situ hybridization, electromobility shift assays and western immunoblot indicate changes in GR and CREB in AI rats. State of the art future approaches to define downstream transcription targets are described.

Omeprazole, clarithromycin and furazolidone for the eradication of Helicobacter pylori in patients with duodenal ulcer.

AIM: To evaluate the efficacy of omeprazole plus clarithromycin and furazolidone in Helicobacter pylori eradication and duodenal ulcer healing in Brazilian patients. METHODS: Forty H. pylori-positive patients with duodenal ulcer were randomized to receive 20 mg omeprazole o.m. or b.d. for 1 month plus 500 mg clarithromycin (b.d. ) and 200 mg furazolidone (b.d.) for 1 week. RESULTS: Three months after the end of the treatment the eradication rates were 90% by intention-to-treat analysis, and 97% by per protocol analysis. Mild side-effects were observed in 25 patients, none of whom abandoned the protocol. No difference was observed between the 20 mg and 40 mg omeprazole daily doses. Cure or significant improvement of the symptoms and of the histological alterations were observed after H. pylori eradication. CONCLUSION: Our results demonstrate that clarithromycin and furazolidone in combination with omeprazole are a good alternative for H. pylori eradication in Brazilian patients with duodenal ulcer.

Epidemiology and etiology of chronic pancreatitis in Brazil: a tale of two cities.

We observed 797 consecutive cases of chronic pancreatitis from 1963 to 1987 in the cities of Belo Horizonte and São Paulo. Alcoholism was the main etiological agent, responsible for 714 cases (89.6%). Chronic calcifying pancreatitis from different etiologies, alcohol, idiopathic, nutritional, and familial, was the most important form of chronic pancreatitis, with 786 cases (98.6%). Only three cases of chronic obstructive pancreatitis were diagnosed. Eight cases of chronic pancreatitis, anatomicopathologically studied, have not showed the typical pancreatic changes of either chronic calcifying pancreatitis or of obstructive pancreatitis and were impossible to classify according to the 1984 Marseilles' classification.

Magnetic resonance cholangiopancreatography in cystic lesions of the pancreas.

There are few reports about magnetic resonance cholangiopancreatography (MRCP) in cystic lesions of the pancreas (KLP). For this reason, we have undertaken a prospective study evaluating the diagnostic efficiency of MRCP as compared with ultrasonography. Twenty-four patients with KLP were examined with magnetic resonance imaging (MRI), and standard and cholangiopancreatography. There were seven cases of cysts associated with acute pancreatitis, 11 patients with KLP and chronic calcifying pancreatitis, five cases of cystic neoplasms of the pancreas, and one polycystic disease of the gland. All cases were first submitted to ultrasonography, which failed to diagnose only a case of cyst associated with chronic pancreatitis depicted by MRCP. We used a GE Signa Horizon 1.5-T system (20 examinations) and a Siemens Magneton Plus 1.5-T machine (four examinations). Eleven patients were operated on. In all cases, it was possible to identify the cysts, the main pancreatic duct and the biliary tree, and verify the relationship of the cyst with neighboring organs. Communication of the cyst with the main pancreatic duct was described in five instances, but we cannot be sure that MRCP would have depicted all cyst-duct communications. The MRI and MRCP images were confirmed by surgery in the 11 operated-on cases. The diagnosis of duct alterations and small pancreatic stones in initial cases of chronic calcifying pancreatitis may be problematic. Clinical findings are very important data to be considered in the differential diagnosis of KLP. Together with the clinical data, MRCP is a very important technique in the diagnostic and therapeutic decision making of KLP. Standard magnetic resonance is advisable as part of the examination in all cases. MRCP is not invasive, is reliable if one knows its limitations, and the patient can return at once to his activities. It allows the analysis of many variables in one examination, contributing to better therapy.

[Prospective study comparing the results obtained by liver puncture biopsy with Chiba fine needle and Menghini needle in some tumors of the liver]. / Estudo prospectivo comparando os resultados obtidos por punção biopsia hepática com agulha fina (Chiba) e de Menghini em alguns tumores do fígado.

Seventy-eight consecutive patients (mean age 59 years) have been submitted to concomitant liver biopsy with Menghini and Chiba needles. The same place in the liver was reached by both needles. In 54 cases there was a clinical suspicion of malignancy and in 24 patients biopsy was indicated by other reasons. In 60 cases the biopsies were oriented by laparoscopy. Comparing the results attained by Menghini needle ("gold standard") and the results of the cytologic study of the material obtained with Chiba needle, the last method has proved to be sensible (92.7%), specific (97.3%) and reliable (accuracy = 94.9%).

[Lithotripsy of gallbladder calculi with extracorporeal shockwaves]. / Litotripsia de cálculos vesiculares por ondas de choque extra-corpóreas.

Two hundred and sixty two patients with gallbladder stones were prospectively evaluated at the Biliary Lithiasis Treatment Unit of the Mater Dei Hospital, Belo Horizonte, MG., and 45 (17.5%) were selected for extracorporal shock wave lithotripsy (ECSWL). From these, 32 were submitted to the procedure. One stone was present in 30 patients, 2 stones in another and 1 patient had 3 stones. The mean diameter was 14.7 mm ranging from 8 to 28 mm. ECSWL was preceded by 1 week course of ursodeoxycholic acid (8 to 10 mg/dk/day) and this medication was continued after the procedure. ECSWL was done with the Lithosthar-Plus apparatus (Siemens). Meperidine (up to 100 mg) IM and pirazolene IV was given when necessary. The intensity of the shock waves was gradually increased to a maximum (9 bar) whenever tolerated. The treatment was well succeeded in 22 cases (71%) with pulverization in 12 (38.7%). In 9 patients (29%) remaining fragments were greater than 4 mm. From these, 3 were submitted to a second session of ECSWL. In 1 patient the stone could not be properly positioned for lithotripsy. The mean number of shock waves was 2,591, ranging from 801 to 4,411. The mean duration of the sessions was 80 min, ranging from 45 to 150 min. In 3 patients, a complete disappearance of fragments was observed in intervals of 1 to 6 months after the procedure. One patient had severe pain during ECSWL and developed acute cholecystitis. One patient had sinus bradycardia. One patient with total stone pulverization, become jaundiced 1 month after ECSWL and a gallbladder carcinoma was found at surgery.(ABSTRACT TRUNCATED AT 250 WORDS)

[Transparieto-hepatic cholangiography with the Chiba needle. Experience in 100 cases]. / Colangiografia transparieto-hepática com agulha de Chiba. Experiência em 100 casos.

One-hundred cases submitted to percutaneous transhepatic cholangiography (PTC) with Chiba's needle were studied. Success rate was 93%, with 100% success in patients with dilated ducts and 78.8% in those with normal ducts. One false-positive result was noticed due to air injection into bile duct in the beginning of the procedure. Complications were seen in 8 cases, without fatalities ascribed to the PTC itself, nor emergency laparotomy was necessary in these cases.

[AW23 and AW24 associated with chronic calcifying pancreatitis of alcoholic origin (author's transl)]. / Associação dos antígenos de histocompatibidade AW23 e AW24 com pancreatite crônica calcificante de origem alcoólica.

Determinations of histocompatibility antigens of A and B loci was undertaken in 22 white male patients with chronic calcifying pancreatitis (CCP) of alcoholic etiology and compared to 100 controls. Results were analysed by the kappa2 test with Yates correction. HLA--AW23 and AW24 were significantly more prevalent in patients than in controls and HLA-B8 was absent in all CCP cases. Although the small number of cases is insufficient to allow any definite conclusion, the obtained data suggests the presence of a genetic predisposition to this disease.

Pancreatic steatorrhea: action of a total lyophilized pancreas preparation: an "in vitro" and "in vivo" study.

A total lyophilized pig pancreas (TLP) preparation was assayed "in vitro" and "in vivo". Lipase activity of the four lots studied was 6.180 +/- 1.122 IU by gram of the commercial product. There was no free trypsin detectable in the preparation. Fifteen pancreatic exocrine insufficiency patients were studied in a period of four weeks. Adult patients received from 35.500 to 35.776 IU of lipase contained in TLP preparation by meal. All patients had their steatorrhea reduced, or normalized, stool weight reduced and body weight increased. There is not an obligatory relation between stool fat loss and stool weight. Steatorrhea decreased at least during the 14 first days of treatment before reaching a stable level. This long delay before reaching a maximum effect deserves to be known: to test the efficiency of a pancreatic extract, the treatment must be given at least two weeks before estimating its results on fecal fat excretion.

[Concomitance of hepatic lesions with chronic calcifying pancreatitis in the alcoholic. Study of 60 cases]. / Concomitância de lesões hepáticas com pancreatite crônica calcificante do alcoólatra. Estudo de 60 casos.

Sixty patients with chronic calcifying pancreatitis (CCP) of alcoholic etiology have been submitted to liver biopsy: in 44 cases the fragment was obtained during surgery, in four cases under laparoscopy, in five during necropsy, in two blind percutaneous biopsy and both surgical and laparoscopic in two cases. Cirrhosis was demonstrated in eight patients (13,3%). The authors emphasises the necessity in patients with alcoholic CCP.