Targeted therapy of cognitive deficits in fragile X syndrome.

Breaking an impasse in finding mechanism-based therapies of neuropsychiatric disorders requires a strategic shift towards alleviating individual symptoms. Here we present a symptom and circuit-specific approach to rescue deficits of reward learning in Fmr1 knockout mice, a model of Fragile X syndrome (FXS), the most common monogenetic cause of inherited mental disability and autism. We use high-throughput, ecologically-relevant automated tests of cognition and social behavior to assess effectiveness of the circuit-targeted injections of designer nanoparticles, loaded with TIMP metalloproteinase inhibitor 1 protein (TIMP-1). Further, to investigate the impact of our therapeutic strategy on neuronal plasticity we perform long-term potentiation recordings and high-resolution electron microscopy. We show that central amygdala-targeted delivery of TIMP-1 designer nanoparticles reverses impaired cognition in Fmr1 knockouts, while having no impact on deficits of social behavior, hence corroborating symptom-specificity of the proposed approach. Moreover, we elucidate the neural correlates of the highly specific behavioral rescue by showing that the applied therapeutic intervention restores functional synaptic plasticity and ultrastructure of neurons in the central amygdala. Thus, we present a targeted, symptom-specific and mechanism-based strategy to remedy cognitive deficits in Fragile X syndrome.

Plant secondary metabolites as potential bioinsecticides? Study of the effects of plant-derived volatile organic compounds on the reproduction and behaviour of the pest beetle Tenebrio molitor.

Modern agriculture has many environmental consequences, such as soil contamination, accumulation of toxic compounds in the environment or risk of adverse effects on nontarget organisms and for these reasons, scientists are seeking a more environmentally friendly alternative to synthetic insecticides. This study investigated the effects of four plant secondary metabolites classified as volatile organic compounds (VOCs), which have potential as bioinsecticides, (E)-2-decenal, furfural, 2-undecanone and (E,E)-2-4-decadienal, in concentrations 10-5 and 10-7 M, on female reproductive processes and larval hatchability of the Tenebrio molitor beetle. Our study indicates proper development of ovaries after application of compounds however the volume of terminal oocytes was significantly reduced, with the strongest effect of (E)- 2-decenal which reduced the volume approximately three times. The relative vitellogenin expression level was reduced, with the strongest effect observed after application of furfural, (E,E)- 2-4-decadienal and (E)- 2-decenal in concentration 10-7 M, at the same time patency index was significantly reduced up to 2-times after application of furfural at 10-7 M. What is more important morphological changes translated into physiological ones. The number of laid eggs was affected, with the strongest inhibition after application of furfural (∼43% reduction), (E,E)- 2-4-decadienal (∼33%) and (E)- 2-decenal at concentration 10-7 M (∼33%). Moreover, we observed up to 13% (in case of 2-undecanone) decrease in larval hatchability. Tested compounds exhibited a repellent effect and caused 60% reduction of insect survivability after (E)- 2-decenal at concentration 10-5 M. Altogether, VOCs seems like potential bioactive compounds in plant protection.

New activity of yamamarin, an insect pentapeptide, on immune system of mealworm, Tenebrio molitor.

In insects, two types of the immune responses, cellular and humoral, constitute a defensive barrier against various parasites and pathogens. In response to pathogens, insects produce a wide range of immune agents that act on pathogens directly, such as cecropins or lysozyme, or indirectly by the stimulation of hemocyte migration or by increasing phenoloxidase (PO) activity. Recently, many new immunologically active substances from insects, such as peptides and polypeptides, have been identified. Nevertheless, in the most cases, their physiological functions are not fully known. One such substance is yamamarin - a pentapeptide isolated from the silk moth Antheraea yamamai. This yamamarin possesses strong antiproliferative properties and is probably involved in diapause regulation. Here, we examined the immunotropic activity of yamamarin by testing its impact on selected functions of the immune system in heterologous bioassays with the beetle Tenebrio molitor, commonly known as a stored grains pest. Our results indicate that the pentapeptide affects the activity of immune processes in the beetle. We show that yamamarin induces changes in both humoral and cellular responses. The yamamarin increases the activity of PO, as well as causes changes in the hemocyte cytoskeleton and stimulates phagocytic activity. We detected an increased number of apoptotic hemocytes, however after the yamamarin injection, no significant variations in the antibacterial activity in the hemolymph were observed. The obtained data suggest that yamamarin could be an important controller of the immune system in T. molitor.

Effects of pea extrusion and enzyme and probiotic supplementation on performance, microbiota activity and biofilm formation in the broiler gastrointestinal tract.

1. The effects of supplementation of broiler chicken diets with pea meal, carbohydrase enzymes and a probiotic were investigated for potential performance improvement. 2. Raw or extruded pea meal (cv Model, grown in Poland) was included in a wheat-soybean meal-based diet at 250 g/kg. The diets were unsupplemented (control) or supplemented with either carbohydrase enzymes (200 U/kg xylanase and 10 U/kg ß-glucanase in feed) or a probiotic (Bacillus subtilis), or both. The diets were fed to Ross 308 broilers aged 9-28 days. 3. After two additional days, chick gastrointestinal tracts were excised and analysed for the presence of Bacillus subtilis biofilm; and the ileal and caecal digesta were analysed for bacterial enzyme activities and to determine the concentration of short-chain fatty acids (SCFAs). 4. Feeding the pea-based diet supplemented with the probiotic compromised feed utilisation, due to higher feed intake. The addition of enzymes to the raw, but not the extruded, pea containing diet partially ameliorated this effect (pea form × additives; P < 0.002). 5. In the ileal digesta, interactions between the dietary treatments were observed for the activities of all bacterial glycolytic enzymes and for SCFA concentrations. ß-glucosidase, α-galactosidase and ß-glucuronidase were highest in birds fed the diet containing extruded pea supplemented with the probiotic and enzymes (pea form x additives; P = 0.018 to P < 0.006). In the caecal digesta, interactions were observed for bacterial enzyme activities, but not for total SCFA concentration. Biofilm formation in the caecum indicated that the probiotic strain was metabolically active in the broiler gut. 6. In conclusion, supplementation of diets containing raw or extruded pea meal with enzymes and a Bacillus subtilis spore-based probiotic modulated microbiota activity but had no clear effects on broiler performance. Probiotic administration did not cause excessive fermentation in the ileum and caecum but enhanced Bacillus subtilis spp. biofilm formation in the caecum, which may be indicative of a beneficial effect on gut health.

Risk of malignant neoplasms in acromegaly: a case-control study.

PURPOSE: Acromegaly is a chronic disease resulting from pathological oversecretion of growth hormone and subsequently insulin growth factor-1. Several complications of the disease have been reported, including cardiovascular diseases, respiratory disorders but also increased risk of benign and malignant neoplasms. The aim of the study was to evaluate the risk of malignant neoplasms in the patients with acromegaly in comparison with the control group. PATIENTS AND METHODS: Medical documentation of acromegalic patients treated in one medical center between 2005 and 2016 has been analyzed. Results were compared with sex- and age-matched group of subjects with prolactinomas and hormonally inactive pituitary lesions hospitalized in the same department. RESULTS: Two hundred patients with acromegaly were included. Control group was composed of 145 patients. Any malignant neoplasm in anamnesis was present in 27 (13.5 %) patients with acromegaly and six (4.1 %) subjects from control group (p = 0.003). Thyroid cancer was present in 14 (7.0 %) patients with acromegaly and two (1.4 %) in control group (p = 0.02). Breast cancer was present in seven women (5.4 % of women) in acromegaly group but none of subjects in control group (p = 0.02). Colon cancer-4 (2.0 %) patients in acromegaly group and 0 in control group (p = 0.14). CONCLUSIONS: Malignant neoplasms are significantly more common in patients with acromegaly. Particularly, risk of thyroid cancer was increased over fivefold. Systematic screening for neoplastic diseases should be important part of follow-up in these patients. Further case-control studies are strongly indicated to evaluate which neoplasms are more common in acromegalic patients and what is the exact risk of malignancy.

Functional homology of tachykinin signalling: The influence of human substance P on the immune system of the mealworm beetle, Tenebrio molitor L.

Tachykinin-related peptides (TRPs) are one of the most prominent families of neuropeptides in the animal kingdom. Insect TRPs display strong structural and functional homology to vertebrate tachykinins (TKs). To study functional homologies between these two neuropeptide families, the influence of human substance P (SP, one of the essential vertebrate TKs) on the immune system of the mealworm beetle, Tenebrio molitor L., was analysed. Human SP influences the phagocytic abilities of T. molitor haemocytes. Peptide injection leads to an increase in the number of haemocytes participating in the phagocytosis of latex beads. In contrast, incubation of haemocytes from non-injected beetles in a solution of physiological saline and SP causes a decrease in phagocytic activity. Treatment with human SP also led to increased adhesion of haemocytes, but no changes in the arrangement of the F-actin cytoskeleton were observed. Interestingly, 6 h after human SP injection, increased DNA integrity in T. molitor haemocytes was reported. The opposite effects were observed 24 h after SP injection. Human SP caused the upregulation of humoral immune responses, such as phenoloxidase (PO) activity in the T. molitor haemolymph, and the downregulation of immune-related genes encoding coleoptericin A, tenecin 3 and Toll receptor. However, genes encoding attacin 2 and cecropin were upregulated. Despite these differences, the antimicrobial activity of T. molitor haemolymph was significantly lower in beetles injected with SP than in control beetles. Moreover, an analysis of the direct influence of SP on lysozyme activity was performed. Our results suggest that SP at a concentration of 10-6 M can directly inhibit lysozyme activity. However, an opposite effect was reported after the application of SP at a concentration of 10-4 M. The presented results suggest structural and functional homology between TK signalling in vertebrates and insects. Primarily, this was visible in the context of the humoral response and general antimicrobial activity of T. molitor haemolymph. However, some of the results related to haemocyte function may also indicate the importance of the TK and TRP sequences for evoking immunological effects.

A possible role of tachykinin-related peptide on an immune system activity of mealworm beetle, Tenebrio molitor L.

Tachykinin-related peptides (TRPs) are important neuropeptides. Here we show that they affect the insect immune system, especially the cellular response. We also identify and predict the sequence and structure of the tachykinin-related peptide receptor (TRPR) and confirm the presence of expression of gene encoding TRPR on Tenebrio molitor haemocytes. After application of the Tenmo-TRP-7 in T. molitor the number of circulating haemocytes increased and the number of haemocytes participating in phagocytosis of latex beads decreased in a dose- and time-dependent fashion. Also, Tenmo-TRP-7 affects the adhesion ability of haemocytes. Six hours after injection of Tenmo-TRP-7, a decrease of haemocyte surface area was observed under both tested Tenmo-TRP-7 concentrations (10-7 and 10-5 M). The opposite effect was reported 24 h after injection, which indicates that the influence of Tenmo-TRP-7 on modulation of haemocyte behaviour differs at different stages of stress response. Tenmo-TRP-7 application also resulted in increased phenoloxidase activity 6 and 24 h after injection. The assessment of DNA integrity of haemocytes showed that the injection of Tenmo-TRP-7 at 10-7 M led to a decrease in DNA damage compared to control individuals. This effect was only visible 6 h after Tenmo-TRP-7 application. After 24 h, Tenmo-TRP-7 injection increased DNA damage. We also confirmed the expression of immune-related genes in nervous tissue of T. molitor. Transcripts for genes encoding receptors participating in pathogen recognition processes and antimicrobial peptides were detected in T. molitor brain, retrocerebral complex and ventral nerve cord. These results may indicate a role of the insect nervous system in pathogen recognition and modulation of immune response similar to vertebrates. Taken together, our results support the notion that tachykinin-related peptides probably play an important role in the regulation of the insect immune system. Moreover, some resemblances with action of tachykinin-related peptides and substance P showed that insects can be potential model organisms for analysis of hormonal regulation of conserved innate immune mechanisms.

Measles virus infection and multiple sclerosis: serological studies.

In 159 patients out of 161 with multiple sclerosis (MS), a significant rise in the level of measles hemagglutination inhibition (HI) antibody was found in the serum and in 92 MS patients the occurrence of measles HI antibody in the CSF was significantly more frequent. MS patients showed CSF humoral response against measles virus by neutralizing test (NV) (76%) more often than by hemagglutination test (37%). CSF FA antibody was found in 60%. In the serum of MS patients the presence of NV, HAd, FA, and GP-RNP was observed. 87% of MS patients showed lowered serum: CSF NV or HI antibody ratios and 78% had a diminished FA antibody ratio. Longitudinal study of serum HI measles virus antibody showed no substantial changes over longer period of the disease. Higher CSF measles antibody titer was found in more disabled patients with a malignant course of the disease (P less than 0.001). It is concluded that either persistent infection with proviruses or nonspecific stimulation of certain clones in individuals with genetic susceptibility provides for an excessive synthesis of humoral viral antibodies in MS.

[Sural nerve phospholipids in some genetically determined nervous system diseases]. / Fosfolipidy w nerwie lydkowym w niektórych genetycznych uwarunkowaniach choroby ukladu nerwowego

Phospholipid fraction pattern was determined in sural nerve in cases of suspected storage diseases. The material was obtained by means fo biopsy from 22 patients. In some patients features of polyneuropathy were found. Normal values of phospholipids were determined in 22 dying patients who had to neurological diseases. The most frequently observed change concerning the biochemical composition of the sural nerve in this material was a fall in the total amount of phosphatidyl ethanolamine (cephalin) and appearance of lysolecithins which are normally absent. In attempted correlation of histological and biochemical findings in the sural nerve with clinical signs of polyneuropathy it was observed that biochemical changes were present in cases without clinical evidence of polyneuropathy. Biochemical as well as histological investigations of sural nerve may help in intravital diagnosis of diseases belonging to the group of thesauroses.

[Patients with multiple sclerosis among the Lódz population. I. Verification of the diagnosis and clinical characteristics]. / Chorzy na stwardnienie rozsiane w populacji Lodzi. I. Weryfikacja rozpoznania i charakterystyka kliniczna.

Home-treated patients with multiple sclerosis who 5-6 years earlier had sought medical advice in neurological health service institutions were examined. Their age was to 46 years; they came from the population of the City Lódz. Slightly over half of them (53%) were patients whose disease had begun at the age of 20-29 years. In 18% of cases the duration of the disease exceeded 14 years. In 60% of cases the disease had a remitting course, in 83% of them multifocal involvement of the nervous system was present. In nearly all cases (97%) pyramidal tract involvement was present, and in 59% the optic nerves were involved. Females prevailed in this group (75%); the proportion of patients aged 40-46 years was higher among the females. In males the onset of the disease before the age of 30 years was significantly more frequent, while in females the proportion of cases beginning at the age of 30-39 years was significantly greater. Among patients aged up to 40 years registered within one year by the neurologists as certain or suspected cases of multiple sclerosis (independently of the duration of the disease) about 7% patients died in a period of 5.7 years on average (10.5% males and 5.3% females). These observations suggest a different onset or course of the disease in both sexes in this population.

[Patients with multiple sclerosis among the Lódz population. II. Clinical course of the disease and patient fitness]. / Chorzy na stwardnienie rozsiane w populacji Lodzi. II. Przebieg choroby i sprawnosc pacjentów.

The motor fitness was assessed in multiple sclerosis patients remaining in their homes in the City of Lódz. The age of the patients was up to 46 years; 5-6 years earlier they had been registered within one year in neurological health service institutions after variously long periods of the disease. The proportion of those with severe motor impairment was 14%, and it was highest (17%) in the groups with disease duration 10-14 years. Nearly half of the group, independently of the age, sex and disease duration, were patients who could move about and perform household works. The proportion of fully fit and nearly fully fit was highest (83%) among those whose disease began under the age of 20 years, and it was lowest (42%) in those with disease onset at the age of 20-29 years. A tendency was observed for a different development of motor function impairment in both sexes depending on the age, age at disease onset and disease duration. In the light of the obtained results it is estimated that in the population of cities in Poland in the whole group of multiple sclerosis patients or subjects with suspected multiple sclerosis registered within one year after variously long disease duration about 40-50% remain after 5-6 years still completely or nearly completely fit.

[A chromatographic method of analysis of mixtures of methylethyl ketone, n-butyl alcohol, xylene, ethylene glycol, ethylene glycol acetate, cyclohexanol, cyclohexanone and butyl glycol vapors in the air]. / Chromatograficzna metoda oznaczania w powietrzu mieszaniny par ketonu metyloetylowego, alkoholu n-butylowego, ksylenu, etyloglikolu, octanu etyloglikolu, cykloheksanolu, cykloheksanonu i butyloglikolu.

A gas chromatographic method for the determination of the mixture of organic solvents in air has been developed. The optimum conditions of the separation of the mixture were obtained in the column packed with 10% UCON HB 2000 coated on Chromosorb W AW DMCS 80-100 mesh. The components of the mixture are collected on activated charcoal and desorbed with CS2. A quantitative analysis is made using methyl isobutyl ketone as an internal standard. The lowest detectable limit under proposed air sampling and analysis conditions is 0.1 TLV (threshold limit value) for each component of the mixture.

[Determination of solvent vapors in the air during use of Butapren OBT III glue]. / Oznaczanie par rozpuszczalników w powietrzu podczas stosowania kleju Butapren OBT-III.

Gas chromatographic method was applied to determine the mixture of extraction benzine, benzene, toluene and ethyl acetate vapours. These are the components of several types of butapren glues. Optimum parameters of separation and estimation of those solvents were obtained with 10% Carbowax 20 M on 80 -- 100 mesh Chromosorb W column. Charcoal adsorption method with CS2 elution was used for air samples collection. In addition, a method was presented for extraction benzine calibration with solvents distilled from OBT-III Butapren glue.

[Determination of lacquer benzine C vapors in the air by means of gas chromatography]. / Oznaczanie w powietrzu par benzyny lakowej C metoda chromatografii gazowej.

A chromatographic method for determining lacquer benzine C in the air has been developed. Appropriate column packing and chromatograph operation parameters have been selected so as to yield summary estimation of all lacquer benzine C components in form of one peak. As a column packing 3% SE-30 has been used deposited on Chromosorb W AW DMCS. This chromatographic method for lacquer benzine C determination is nonspecific when related to xylene and toluene.

[Determination of petroleum and benzene vapors in the air by gas chromatography]. / Oznaczanie w powietrzu par benzyny ekstrakcyjnej i benzenu metoda chromatografii gazowej.

A gas chromatographic method for determination of extraction naphtha and benzene has been developed. The best parameters of chromatographic separation were found. Under those conditions it is possible to estimate the total concentration of all ingredients of extraction naphtha as one peak clearly separated from benzene peak. Two methods of calibration curve preparation were considered. One of them consists in preparation of calibration curve from extraction naphtha standard solutions. The other one consists in preparation of calibration curve from standard extraction naphtha vapours adsorbed on charcoal.

Micro-imaging of implanted scaffolds using combined MRI and micro-CT.

RATIONALE AND OBJECTIVES: Thanks to the advanced studies in biomaterial engineering a panoply of polymers can be used to manufacture porous scaffolds for bone tissue regeneration. Suitability of the scaffold for its purpose is determined by factors like size of the pores, its orientation and shape, as well as biocompatibility of the material. Even though a variety of analysis methods is available for in vitro studies, investigating the process of bone reconstruction on implanted scaffold meets with difficulties. METHODS AND MATERIALS: Polylactide porous sponges imbued in hydroxyapatite were implanted into long bones of white New Zealand rabbits for 3 months. The bones obtained from the animals were subjected to MRI and µCT imaging. The obtained images were subsequently fused together. RESULTS AND CONCLUSIONS: Combined MRI and µCT resulted in high resolution diagnostic images which allow for: implant positioning, inflammation divulgement, rating degree of implant resorption, observation of newly formed trabeculae, texture analysis and other quantitative measurements.