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Single-cell transcriptomics methods have become very popular to study the cellular composition of organs and tissues and characterize the expression profiles of the individual cells that compose them. The main critical step in single-cell transcriptomics is sample preparation. Several methods have been developed to preserve cells after sample dissociation to uncouple sample handling from library preparation. Yet, the suitability of these methods depends on the types of cells to be processed. In this project, we perform a systematic comparison of preservation methods for droplet-based single-cell RNA-seq (scRNA-seq) on neural and glial cells derived from induced pluripotent stem cells (iPSCs) and highlight their strengths and weaknesses. We compared the cellular composition and expression profile of single-cell suspensions from fresh NPCs with that of NPCs preserved with Dimethyl Sulfoxide (DMSO), Methanol, vivoPHIX and Acetil-methanol (ACME). Our results show that while DMSO provides the highest cell quality in terms of RNA molecules and genes detected per cell, it strongly affects the cellular composition and the expression profile of the resulting datasets. In contrast, methanol fixed samples display a cellular composition like that of fresh samples while providing a good cell quality and smaller expression biases. Taken together, our results show that methanol fixation is the method of choice for performing droplet-based single-cell transcriptomics experiments on neural cell populations.

Lettuce (Lactuca spp.) is an annual and self-pollinating crop that belongs to the Asteraceae (Compositae) family. It is one of the most globally essential commercial vegetable crops, used in salads and sandwiches. The lettuce leaves are used to make a cigarette without nicotine. Seeds and stems contain edible oil and dried latex. Gene banks have conserved a large pool of lettuce's genetic resources, including wild Lactuca species with the same chromosome numbers 2n = 2x = 18. Lactuca species vary greatly in terms of geographical distribution and morpho-agronomic characteristics. By crossing commercial varieties with locally adapted varieties, novel alleles can be introduced, increasing genetic diversity and making preselection for desirable traits easier. Lettuce breeders and geneticists' main objectives are to improve lettuce for various desirable traits, including tolerance to abiotic and biotic stress and high yield. These targets accomplished with modern genomics tools together with traditional breeding methods. This chapter discusses lettuce conservation and biodiversity, stages of lettuce breeding, agriculture practices, and conventional breeding techniques and their restrictions. It also includes modern plant breeding tools and marker-assisted breeding, editing of genome, and genetic engineering.

Little is known about the interactions among phagocytes and antifungal agents and the antifungal immunomodulatory activities on

Enterococcus faecalis and Candida albicans are two of the most significant opportunistic and intricate nosocomial pathogens. They reside overlapping niches as major constituents of the gastrointestinal (GI) and oral microbiome. This study was designed to investigate the interaction between these two opportunistic pathogens that could affect treatment strategies and influence the interkingdom signaling and sensing in the microbiome conceptions. E. faecalis was isolated from 150 samples collected from human urine, cattle raw milk, milk products and chicken intestine and identified by both the routine microbiological and molecular identification methods based on 16S rRNA gene. Isolates susceptibility to nine antimicrobials was analyzed using the disk-diffusion method and E. faecalis isolates showed multidrug resistance. Most of C. albicans isolates were able to form biofilm with different intensities. Cell-free supernatant (CFS) of E. faecalis was purified and tested against C. albicans and/or biofilm formation.It inhibited the growth of C. albicans and their filamentation with regards to biofilm formation in vitro. The data presented here demonstrated the potential antifungal activity of E. faecalis CFS and could explain the biological interaction between these two microbes in one hand and the possibility of using E. faecalis CFS as a biological product to control yeast infection in the other hand.

Many software implementations for the schemes used for public key cryptosystems, of which elliptic curve cryptography is the most powerful, has been concerned with performance and efficiency. However, the advent of side channel attacks with their diverse categories, such as timing, fault and power analysis attacks, impose us to reconsider new strategies in implementing more secured elliptic curve algorithms to thwart any information leakage that leads to break the security of these algorithms. In this paper, we propose a new optimization on the algorithmic level for computing the arithmetic of elliptic curve point over prime fields to countermeasure side channel attacks that threaten elliptic curve cryptosystems. Indeed, these attacks present nowadays a realistic threat to cryptographic applications and have been proved to be very effective against most cryptosystems. To develop a digital signature algorithm that targets performance and security against side channel attacks, we built a library for the underlying prime fields arithmetic for the common fields specialized by NIST and SECG. Our work can be employed in numerous applications such as, E-health, E-banking, E-commerce and E-governance.

l-Arginine deiminase (ADI) has a powerful anticancer activity against various tumors, via arginine depletion, arresting the cell cycle at G1 phase. However, the current clinically tried bacterial ADI displayed a higher antigenicity and lower thermal stability. Thus, our objective was to purify and characterize this enzyme from thermophilic fungi, to explore its catalytic and antigenic properties for therapeutic uses. ADI was purified from thermophilic Aspergillus fumigatus KJ434941 to its electrophoretic homogeneity by 5.1-fold, with molecular subunit 50 kDa. The purified ADI was PEGylated and covalently immobilized on dextran to explore its catalytic properties. The specific activity of free ADI, PEG-ADI, and Dex-ADI was 26.7, 21.5, and 18.0 U/mg, respectively. At 50°C, PEG-ADI displays twofold resistance to thermal denaturation (t1/2 13.9 h), than free ADI (t1/2 6.9 h), while at 70°C, the thermal stability of PEG-ADI was increased by 1.7-fold, with similar stability to Dex-ADI with the free one. Kinetically, free ADI had the higher catalytic affinity to arginine, followed by PEG-ADI and Dex-ADI. Upon proteolysis for 30 min, the residual activity of native ADI, PEG-ADI, and Dex-AD was 8.0, 32.0, and 20.0% for proteinase K and 10.0, 52.0, and 90.0% for acid protease, respectively. The anticancer activity of the ADIs was assessed against HCT, HEP-G2, and MCF7, in vitro. The free and PEG-ADI exhibits a similar cytotoxic efficacy for the tested cells, lower than Dex-ADI. The free ADI had IC50 value 22.0, 16.6, and 13.9 U/mL, while Dex-ADI had 3.98, 5.18, and 4.43 U/mL for HCT, MCF7, and HEPG-2, respectively. The in vitro anticancer activity of ADI against HCT, MCF7, and HEPG-2 was increased by five-, three-, and threefold upon covalent modification by dextran. The biochemical and hematological parameters of the experimented animals were not affected by ADIs dosing, with no signs of anti-ADI immunoglobulins in vivo. The in vivo half-life time of free ADI, PEG-ADI, and Dex-ADI was 29.7, 91.1, 59.6 h, respectively. The present findings explored a novel thermostable, less antigenic ADI from thermophilic A. fumigatus, with further molecular and crystallographic analyses, this enzyme will be a powerful candidate for clinical trials.

This investigation was designed to throw light on the contamination rate of meat and meat products with mould and aflatoxins. The prevalence and population density of the mycobiota of 350 beef meat and meat product samples (frozen meat, minced meat, liver, kidney, luncheon, sausage, hawawchi), 50 of each, collected from different abattoirs, markets and shopkeepers in Egypt were studied. The highest total mould count/g was obtained from the sausage samples (4.20 ± 1.25 X 104), whereas the frozen meat samples yielded the lowest mould count (2.00 ± 1.2 X 102). The most frequently encountered mould genera from the examined samples were Aspergillus and penicillium. Aspergillus species were isolated at percent of 100%, 80%, 80%, 78%, 74%, 70% and 50% from minced meat, hawawchi, sausage, luncheon, liver, frozen meat and kidney samples respectively. Aspergillus flavus was the most predominant isolated species. All A. parasiticus and 77.96% of A. flavus isolates were positive for aflatoxin production on coconut agar medium (CAM), thin layer chromatography (TLC) and fluorometric immune affinity method. Detection of aflatoxin residues in meat and meat products samples revealed that the highest mean values of aflatoxin residues (µg/kg) B1, B2, G1 and G2 were detected in the kidney samples (12.36 ± 1.89, 9.84 ± 1.63, 5.38 ± 1.36 and 6.84 ± 1.39, respectively)followed by the liver (13.81 ± 1.96, 3.26 ± 0.92, 2.51 ± 0.63 and 1.36 ± 0.38), luncheon (3.71 ± 1.35, 3.59± 1.12, 5.24 ± 1.12 and 6.77 ±1.49), hawawchi (11.03 ± 2.43, 2.25± 0.52, 2.54 ± 0.99 and 2.56 ± 0.27), minced meat (3.62 ± 0.88, 3.40 ± 0.82, 4.24 ± 0.85, 2.83 ± 0.60) and frozen meat samples which had the lowest level of AFs (4.80 ± 0.89, 5.3 ± 2.1, 1.71 ± 0.60 and 0.0) respectively. The detected levels of aflatoxin residues in the present samples were compared with the international permissible limits of WHO, FAO and FDA. Most of detected aflatoxins levels in meat and meat product samples were more than the permissible limits thus would be unfit for human consumption. The present study attracts the attention to potential risk for aflatoxin producing moulds and aflatoxins contamination and strongly recommends reduction in various causes of contamination as lack of hygienic measures during slaughtering, handling, transportation, storage or processing to meat and its products and also avoiding contaminated spices used in meat processing

Rapid and accurate identification of yeast is increasingly important to stipulate the appropriate therapy thus reducing morbidity and mortality related to yeast infections. Vibrational spectroscopic techniques (infrared (IR) and Raman) could provide potential alternatives to conventional typing methods, because they constitute a rapid, inexpensive and highly specific spectroscopic fingerprint through-which microorganism can be identified. The present study evaluate (FTIR) spectroscopy as a sensitive and effective assay for the identification of the most frequent yeast species isolated from human and animals. One hundred and twenty-eight yeasts isolated from infected human mouths/vaginas, chronic diseased cows, crop mycosis in chicken and soil contaminated with pigeon droppings were phenotypically identified. Using universal primers, ITS1/ITS4, we have amplified ITS1-5.8S-ITS2 rDNA regions for 39 yeast isolates as representative samples. The PCR products were digested with restriction enzyme MspI and examined by PCR-RFLP, which was an efficient technique for identification of Candida spp., Cryptococcus neoformans and Trichosporon asahii. Further, identification of the same 39 isolates were done by FTIR spectroscopy and considered as reference for other strains by comparison of their FTIR spectra. The current study has sharply demonstrated the significant spectral differences between the various examined species of Candida, Cryptococcus, Trichosporon, Rhodotorula and Geotrichum isolated from different sources. Decisively, our research has confirmed that FTIR spectroscopy is a promising diagnostic tool, because of its sensitivity, rapidity, high differentiation capacity and simplicity compared to conventional/molecular techniques.

In this work, two new low area designs on FPGA for a scalable elliptic curve point multiplication are presented. The first design is a pure software implementation on a compact 8-bit embedded microcontroller, the PicoBlaze from Xilinx. The second is a novel hardware architecture for a compact ECC processor. Both designs are scalable across the curves m=113,131,163,193 recommended by SECG and consume respectively only (23%) and (61%) of the available resources on the smallest and lowest cost Spartan-III XC3S50 device. The proposed designs are believed to be the first smallest reported designs to date that support scalability with low area for a suite of curves recommended by an ECC standard.

Objective To evaluate the possible association of Parkinson disease (PD) and melanoma in North America. Design, Setting, and Patients Thirty-one centers enrolled patients with idiopathic PD. At visit 1, a neurologist obtained a medical history. At visit 2, a dermatologist recorded melanoma risk factors, performed a whole-body examination, and performed a biopsy of lesions suggestive of melanoma for evaluation by a central dermatopathology laboratory. We compared overall prevalence of melanoma with prevalence calculated from the US Surveillance Epidemiology and End Results (SEER) cancer database and the American Academy of Dermatology skin cancer screening programs. Results A total of 2106 patients (mean [SD] age, 68.6 [10.6] years; duration of PD, 7.1 [5.7] years) completed the study. Most (84.8%) had received levodopa. Dermatology examinations revealed 346 pigmented lesions; dermatopathological findings confirmed 20 in situ melanomas (0.9%) and 4 invasive melanomas (0.2%). In addition, histories revealed 68 prior melanomas (3.2%). Prevalence (5-year limited duration) of invasive malignant melanoma in the US cohort of patients with PD (n = 1692) was 2.24-fold higher (95% confidence interval, 1.21-4.17) than expected in age- and sex-matched populations in the US SEER database. Age- or sex-adjusted relative risk of any melanoma for US patients was more than 7 times that expected from confirmed cases in American Academy of Dermatology skin cancer screening programs. Conclusions Melanoma prevalence appears to be higher in patients with PD than in the general population. Despite difficulties in comparing other databases with this study population, the study supports increased melanoma screening in patients with PD.

Grimes, J.D., Hassan, M.N., Thakar, J.H.: Prevention of Progression of Parkinson's Disease with Antioxidative Therapy. Prog. Neuro. Psychopharmacol. & Biol. Psychiat. 1988, 12 :165–172 1. 1. Oxidative mechanisms in dopaminergic neurons may contribute to cell death and the progression of Parkinson's Disease. 2. 2. The free radical auto-toxicity concept has scientific evidence to support it. 3. 3. Clinical trials are underway to assess the protective effect of augmenting the free radical scavenging system with vitamin E and inhibiting catecholamine oxidation with deprenyl.

tran-3-(3,4-Dihyroxyphenyl)-4-methyl-1-(n-propyl)pyrrolidine 13 trans-3-(3-Hydroxyphenyl)-4-methyl-1(n-propy)pyrrolidine 14 were synthesized and evaluated for dopaminergic activity. The stereochemical assignments of 13 and 14 were determined by nmr. Both 13 and 14 were either inactive or weakly active in most dopaminergic tests.

SUMMARYUsing a method of a gradual increase of bromocriptine with a concomitant reduction of Sinemet® (levodopa 250 mg + carhidopa 25 mg), 19 patients with advanced Parkinson’s disease have been treated for periods of up to 22 months and 16 of them have shown improvements of varying degrees. Eighteen patients were able to tolerate bromocriptine addition, with early transient adverse effects occurring in seven cases. In contrast to several previously reported studies, it was found necessary to withdraw bromocriptine in only one case.With the drugs currently available, bromocriptine has a role in the management of patients with advanced Parkinson’s disease. The method described here may allow a greater number of patients to be given a trial with this drug.

The occurrence of adverse extrapyramidal effects following metoclopramide (MCP) therapy has been well documented in humans. In rats, MCP produced catalepsy and inhibited apomorphine-induced stereotypy, locomotor activity, and rotational behavior. MCP also accelerated dopamine turnover, potently stimulated prolactin release, and, at high concentrations, inhibited [3H]spiperone binding to striatal membranes. These behavioral and biochemical effects induced by MCP were similar to those observed with haloperidol. We conclude that MCP possesses most of the pharmacological properties of neuroleptic agents.

Concurrent administration of lithium (Li) significantly attenuates the dopamine (DA) depleting effects of reserpine and tetrabenazine, but does not change alpha-methyl-p-tyrosine (AMPT) induced DA depletion in rat brain. This effect of Li is probably mediated, in part, by inhibiting the magnesium-dependent binding of both reserpine and tetrabenazine to their specific receptor sites. Such interaction between these drugs may attenuate the beneficial effects of tetrabenazine and reserpine on patients with tardive dyskinesia or tardive dystonia who are treated concurrently with lithium for their psychiatric disorder.

Spiperone dithiocarbamate (SPDC) was prepared by reacting spiperone with carbon disulfide followed by sodium hydroxide. SPDC was labelled with 99m Tc by reduction of pertechnetate with formamidine sulfinic acid or sodium dithionite at alkaline pH, resulting in ∼40% incorporation of 99m Tc. The lipophilic complex was conveniently isolated at high specific activity and high radiochemical purity by extraction into dichloromethane, which was then evaporated and the residue was redissolved in a 1:3 mixture of ethanol and saline containing 0.1 mg/mL gentisic acid. Biodistribution studies following i.p. injection in rats showed low uptake of radioactivity in the brain, but striatum/cortex and striatum/cerebellum ratios were reduced by pretreatment with haloperidol. This agent may allow imaging of dopamine D-2 receptors using single-photon emission computed tomography (SPECT).

Long-term lithium treatment attenuated the hypokinetic effect of reserpine in a patient with tardive dyskinesia. In rats, prophylactic lithium administration inhibits reserpine-induced dopamine depletion in the brain. These data indicate that lithium may limit the therapeutic efficacy of reserpine in tardive dyskinesia.

The effects of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), its metabolite 1-methyl-4-phenyl pyridinium ion (MPP+, cyperquat) and a structurally-related compound paraquat on mitochondrial functions were investigated in isolated organelles from rat striatum, cortex and liver. MPTP (0.1-1.0 mM) had no significant effect on various parameters of mitochondrial oxidative phosphorylation. In contrast, MPP+ (0.5 mM) inhibited the oxidation of the nicotinamide adenine dinucleotide (NAD+)-linked substrates pyruvate and malate but not that of the flavin adenine dinucleotide (FAD+)-linked substrate succinate. Paraquat (5.0 mM) significantly stimulated basal oxygen consumption (state 4) without influencing the oxygen utilization (state 3) associated with adenosine diphosphate (ADP) phosphorylation. Thus, these structurally-related compounds have different effects on mitochondrial oxidative phosphorylation, but the organelles from striatum, cortex and liver were affected in a similar manner by these compounds.

Chronic treatment of rotating rats with equipotent doses of the dopamine (DA) agonists apomorphine (APO), 3-(3,4-dihydroxyphenyl)-1-n-propylpyrrolidine hydrobromide (DPPP) and bromocriptine (BRO) for four weeks resulted in marked differences in rotational activity following acute administration of these agonists. Whereas chronic treatment with APO and DPPP failed to produce any significant changes in agonist-induced rotational behavior, chronic BRO treatment induced a progressive increase in rotational activity up to a mean 200% increase over controls at four weeks. These findings may, in part, explain the long-term clinical efficacy of bromocriptine in patients with Parkinson's disease.

3-(3,4-Dihydroxyphenyl)-1-n-propylpyrrolidine hydrobromide was synthesized by a six-step reaction sequence and was evaluated, and compared with apomorphine, for central dopaminergic agonist activity. The compound produced behavioral and biochemical changes characteristic of central dopaminergic stimulation. Administration of 3-(3,4-dihydroxyphenyl)-1-n-propylpyrrolidine hydrobromide to rats resulted in the reversal of the reserpine syndrome, stereotypic behavior, contralateral turning following 6-hydroxydopamine lesion of the substantia nigra, a decrease in dopamine turnover, and inhibition of prolactin release. These results indicate that 3-(3,4-dihydroxyphenyl)-1-n-propylpyrrolidine hydrobromide is a dopaminergic agonist. However, the compound exhibited a lower potency but a slightly longer duration of action than apomorphine.