Detection of Neisseria meningitidis and unknown Gammaproteobacteria in cerebrospinal fluid using the two-step universal method

Bacterial meningitis is insufficiently diagnosed based on microscopic, cultural, and multiplex-polymerase chain reaction (M-PCR). The use of already established universal method (UM) offers the ultimate solution to the detection and potential identification of bacteria in cerebrospinal fluid (CSF) samples. We have applied the UM together with a newly established Anchored Multiplex PCR (AMD4; Streptococcus pneumoniae, Haemophilus influenzae, Neisseria meningitidis, and Listeria monocytogenes) to screen 130 CSF samples obtained from suspected meningitis cases for any bacterium. This two-stage UM approach was able to show that three of the samples contained bacterial DNA, only one of the three samples (K1) was shown to contain N. meningitidis whereas the other two samples (A35 and H1) were negative with AMD4. Nucleotide sequencing and BLAST analyses of 16S amplicons obtained by the UM from samples A35 and H1 showed no significant homology (<90%) to any available 16S sequence, yet indicated both bacteria (A35 and H1) to share 94.2% similarity. Both bacteria belonged to Gammaproteobacteria. The bacterium from sample K1 was isolated by culture and identified as N. meningitidis. The other two samples were negative by culture according to the clinical laboratories at both hospitals; A35 was from a patient who had received empirical antimicrobial therapy prior to sample collection. The remaining 127 samples were shown by the UM to be negative in accordance with clinical and laboratory findings. The UM can contribute significantly to the identification of bacterial meningitis cases to initiate empirical antimicrobial therapy within 3 h of sample collection. Simultaneously, bacterial meningitis can be ruled out from samples producing negative UM results. AMD4 application will detect and identify the major pathogens of bacterial meningitis whereas the UM will detect any bacterium, UM can potentially identify any bacterium as long as it is represented in the nucleotide databases; if not represented, it is labeled as unknown. We recommend the utilization of the UM in clinical testing; we also recommend culturing, characterization and identification of these unknown bacterial agents of meningitis as well as others.We would like to thank the Administrations, Doctors, and Nurses and meningitis patients of The Alia Hospital, Al-Ahli Hospital, and The Caritas Hospital, West Bank, Palestine for their unwavering support and help in collecting CSF samples used in this study

Synthesis, physicochemical, conformation and quantum calculation of novel N-(1-(4-bromothiophen-2-yl)ethylidene)-2-(piperazin-1-yl)ethanamine Schiff base

N-(1-(4-bromothiophen-2-yl)ethylidene)-2-(piperazin-1-yl)ethanamine Schiff base ligand was prepared in very good yield by condensation of equimolar amounts of 1-(4-bromothiophen-2-yl)ethanone with 2-(piperazin-1-yl)ethanamine under reflux condition using alcohol media. The desired Schiff base was analyzed on the basis of its MS, elemental analysis, UV-visible, FT-IR and NMR analysis. The E and Z optimization was performed to figure out the most stable isomer. Several DFT quantum calculation like: TD-SCF, MPE, IR-vibration, NMR, Mulliken population were carried out by B3LYP level of theory. The experimental analyses of the compound were compared to their theoretical coordinates

The Epimmunity Theory: The Single Cell Defenses against Infectious and Genetic Diseases

Single cell defense against diseases defines “epimmunity.” Epimmunity is complementary to the immune system and can neither be substituted by innate nor by acquired immunity. Epimmunity, the proposed new branch of immunity, is further explored and analyzed for enucleated mature mammalian erythrocytes and nucleated erythrocytes of non-mammalian vertebrates leading to the development of “The Epimmunity Theory.” Enucleation of mammalian erythroblast and inactivation of nuclei in erythrocytes of non-mammalian vertebrates are major contributors to the collective immunity: epimmunity, innate, and acquired. The fact that diseases of mature erythrocytes (MEs) are rare supports the notion that a single cell can resist microbial and genetic diseases; MEs are refractory to malaria and cancer. Nucleated cells, such as B-cells, T-cells, hepatocytes, and cell developmental stages are susceptible to genetic and specific microbial diseases depending on their nuclear activities and the receptors they express; such cells show lower epimmunity relative to MEs. Epimmunity is important as a disease insulator that prevents the spread of diseases from an infected tissue to the majority of other tissues. Breakdown of epimmunity may lead to disease development

Ultrasound-assisted synthesis of two novel CuBr(diamine)2·H2OBr complexes: Solvatochromism, crystal structure, physicochemical, hirshfeld surface thermal, DNA/binding, antitumor and antibacterial activities

Two new hydrated monocationic Cu(II) complexes with 1,3-propylenediamine and 1,2-ethylenediamine of general formula CuBr(N-N)2·H2OBr were prepared. The complexes were identified by means of several spectroscopic tools (Uv-visible, IR and MS), thermally (TG/DTA) and CHN-elemental analysis. The three dimensional structure for complex A and B was provide by X-ray diffraction studies and showed the Cu(II) ion as 4 + 1 + 1 coordinated, four nitrogen atoms of the diamine ligands, one bromide ion and one H2O semi-coordinated to the Cu(II) center, a typical trans effect is clearly observed in the two complexes. The molecular crystal structures are linked via several H-bonds like N_H…Br and N_H…O. Additionally, intra-molecular H-bonds of kind C_H…Br is observed; these interactions lead to crystal structure three dimensional architecture packing. Hirshfeld surfaces (HSA) analysis was served to figure out the inter-contacts and fingerprints atoms percentage. DNA-binding, antitumor and antibacterial effectiveness of the desired complexes were evaluated