Cold Ethanol Research Articles

Crystal Structure of Human Fibrinogen

A crystal structure of human fibrinogen has been determined at approximately 3.3 A resolution. The protein was purified from human blood plasma, first by a cold ethanol precipitation procedure and then by stepwise chromatography on DEAE-cellulose. A product was obtained that was homogeneous on SDS-polyacrylamide gels. Nonetheless, when individual crystals used for X-ray diffraction were examined by SDS gel electrophoresis after data collection, two species of alpha chain were present, indicating that some proteolysis had occurred during the course of operations. Amino-terminal sequencing on post-X-ray crystals showed mostly intact native alpha- and gamma-chain sequences (the native beta chain is blocked). The overall structure differs from that of a native fibrinogen from chicken blood and those reported for a partially proteolyzed bovine fibrinogen in the nature of twist in the coiled-coil regions, likely due to weak forces imparted by unique crystal packing. As such, the structure adds to the inventory of possible conformations that may occur in solution. Other features include a novel interface with an antiparallel arrangement of beta chains and a unique tangential association of coiled coils from neighboring molecules. The carbohydrate groups attached to beta chains are unusually prominent, the full sweep of 11 sugar residues being positioned. As was the case for native chicken fibrinogen, no resolvable electron density could be associated with alphaC domains.

UV Radiation Regulates Mi-2 through Protein Translation and Stability

Dermatomyositis (DM) is an autoimmune disease, which is often accompanied by the development of disease-specific autoantibodies directed against the SNF2-superfamily helicase, Mi-2. Recent evidence suggests that ultraviolet radiation exposure may be an important risk factor for the development of not only the disease but also specific autoimmunity against Mi-2. Consequently, we investigated the effects of ultraviolet radiation on Mi-2 protein expression. We observed an increase in protein levels upon ultraviolet radiation exposure in cell culture systems. These changes in expression occur quite rapidly, are maximized just 1 h following exposure, and are unique to Mi-2 when compared with other members of the NuRD complex. Changes in protein levels are not mediated through transcriptional mechanisms. Treatment results in a more efficiently translated message through regulatory elements in the 5'-UTR region of the transcript. Investigation into protein half-life further demonstrated increased stability of Mi-2 following UV exposure. Taken together, we describe a system by which Mi-2 protein expression can be quickly increased following UV exposure and then maintained up to 16 h later. These data provide a novel regulation of an important transcriptional regulator and provide insight into the possible mechanisms of the development of DM and associated autoantibodies.

The TREX1 Double-stranded DNA Degradation Activity Is Defective in Dominant Mutations Associated with Autoimmune Disease

Mutations in TREX1 have been linked to a spectrum of human autoimmune diseases including Aicardi-Goutières syndrome (AGS), familial chilblain lupus (FCL), systemic lupus erythematosus, and retinal vasculopathy and cerebral leukodystrophy. A common feature in these conditions is the frequent detection of antibodies to double-stranded DNA (dsDNA). TREX1 participates in a cell death process implicating this major 3' --> 5' exonuclease in genomic DNA degradation to minimize potential immune activation by persistent self DNA. The TREX1 D200N and D18N dominant heterozygous mutations were identified in AGS and FCL, respectively. TREX1 enzymes containing the D200N and D18N mutations were compared using nicked dsDNA and single-stranded DNA (ssDNA) degradation assays. The TREX1WT/D200N and TREX1WT/D18N heterodimers are completely deficient at degrading dsDNA and degrade ssDNA at an expected approximately 2-fold lower rate than TREX1WT enzyme. Further, the D200N- and D18N-containing TREX1 homo- and heterodimers inhibit the dsDNA degradation activity of TREX1WT enzyme, providing a likely explanation for the dominant phenotype of these TREX1 mutant alleles in AGS and FCL. By comparison, the TREX1 R114H homozygous mutation causes AGS and is found as a heterozygous mutation in systemic lupus erythematosus. The TREX1R114H/R114H homodimer has dysfunctional dsDNA and ssDNA degradation activities and does not detectibly inhibit the TREX1WT enzyme, whereas the TREX1WT/R114H heterodimer has a functional dsDNA degradation activity, supporting the recessive genetics of TREX1 R114H in AGS. The dysfunctional dsDNA degradation activities of these disease-related TREX1 mutants could account for persistent dsDNA from dying cells leading to an aberrant immune response in these clinically related disorders.

Microbial community analysis of a full-scale membrane bioreactor treating industrial wastewater

A Kubotatrade mark submerged membrane bio-reactor was applied to treat wastewater from a sugar manufacturing industry. To achieve optimal results, fundamental and extended understanding of the microbiology is important. Fluorescence in situ hybridization was used to evaluate the microbial community present. The majority of cells visualized in the sludge flocs by staining with the DNA fluorochrome DAPI, hybridized strongly with a bacterial probe. Probes specific for the alpha-, beta-, and gamma-subclasses of proteobacteria and high G + C Gram positive bacteria were used to characterize the community structures by in situ hybridization. Sampling was carried out over 12 weeks and samples were fixed with 4% paraformaldehyde for gram positive organisms and ice cold ethanol for gram negative organisms. The activated sludge population usually constitutes about 80 to 90% of proteobacteria. However, in this study it was found that a relatively small amount of proteobacteria was present within the system. No positive hybridization signal was observed with any of the applied eubacterial family- level probes.

Ets2 Maintains hTERT Gene Expression and Breast Cancer Cell Proliferation by Interacting with c-Myc

Human telomerase reverse transcriptase (hTERT) underlies cancer cell immortalization, and the expression of hTERT is regulated strictly at the gene transcription. Here, we report that transcription factor Ets2 is required for hTERT gene expression and breast cancer cell proliferation. Silencing Ets2 induces a decrease of hTERT gene expression and increase in human breast cancer cell death. Reconstitution with recombinant hTERT rescues the apoptosis induced by Ets2 depression. In vitro and in vivo analyses show that Ets2 binds to the EtsA and EtsB DNA motifs on the hTERT gene promoter. Mutation of either Ets2 binding site reduces the hTERT promoter transcriptional activity. Moreover, Ets2 forms a complex with c-Myc as demonstrated by co-immunoprecipitation and glutathione S-transferase pulldown assays. Immunological depletion of Ets2, or mutation of the EtsA DNA motif, disables c-Myc binding to the E-box, whereas removal of c-Myc or mutation of the E-box also compromises Ets2 binding to EtsA. Thus, hTERT gene expression is maintained by a mechanism involving Ets2 interactions with the c-Myc transcription factor and the hTERT gene promoter, a protein-DNA complex critical for hTERT gene expression and breast cancer cell proliferation.

Clearance of dengue virus in the plasma‐derived therapeutic proteins

Viral safety is of paramount importance for human plasma-derived therapeutic proteins. Recent reports of blood-associated transmission and continuous regional outbreaks of dengue fever have prompted a validation of clearance of dengue virus in the manufacture processes of the plasma-derived products. A high titer of cultured dengue virus serotype 2 was spiked into process samples before individual steps of albumin and immunoglobulin manufacture processes, including cold ethanol precipitation, cation-exchange chromatography, pasteurization, solvent/detergent treatment, and virus filtration. Clearance of dengue virus was quantified with TCID(50) assays in the culture of Vero E6 cells and, when appropriate, real-time polymerase chain reaction (RT-PCR) assays. The individual process steps were all effective in the inactivation and/or removal of dengue virus, and the data obtained clearly demonstrate that the risk of dengue virus transmission was reduced cumulatively by at least 10.12 and at least 14.24 log in the albumin and immunoglobulin manufacture processes, respectively. The dedicated viral inactivation and/or removal approaches currently implemented in the manufacture of plasma-derived products provide a good safety margin with regard to the transmission of dengue virus.

The PDZ Protein Mupp1 Promotes Gi Coupling and Signaling of the Mt1 Melatonin Receptor

Intracellular signaling events are often organized around PDZ (PSD-95/Drosophila Disc large/ZO-1 homology) domain-containing scaffolding proteins. The ubiquitously expressed multi-PDZ protein MUPP1, which is composed of 13 PDZ domains, has been shown to interact with multiple viral and cellular proteins and to play important roles in receptor targeting and trafficking. In this study, we show that MUPP1 binds to the G protein-coupled MT(1) melatonin receptor and directly regulates its G(i)-dependent signal transduction. Structural determinants involved in this interaction are the PDZ10 domain of MUPP1 and the valine of the canonical class III PDZ domain binding motif DSV of the MT(1) carboxyl terminus. This high affinity interaction (K(d) approximately 4 nm), which is independent of MT(1) activation, occurs in the ovine pars tuberalis of the pituitary expressing both proteins endogenously. Although the disruption of the MT(1)/MUPP1 interaction has no effect on the subcellular localization, trafficking, or degradation of MT(1), it destabilizes the interaction between MT(1) and G(i) and abolishes G(i)-mediated signaling of MT(1). Our findings highlight a previously unappreciated role of PDZ proteins in promoting G protein coupling to receptors.

Process development and pilot-scale production of therapeutic antisera against botulinum toxin type A and B

This study aimed at process development and pilot-scale production of botulinum antitoxin for an urgent botulism-poisoning patient, contributing to steady antitoxin supply for bio-terrorism. After horses were immunized with formalin-inactivated botulinum toxin type A and B, the sera were prepared. Following purifications of IgG fractions by cold ethanol fractionation, caprylate fractionation, anion and exchange chromatography, and F(ab′)2 by cold ethanol fractionation, pepsin digestion, and cation exchange chromatography, the specificity and protective efficacy against botulinum toxin type A and B were analyzed by enzyme-linked immunosorbent assay (ELISA), Western blot, and mouse neutralization assay. Based on the result of mouse neutralization assay, it was found that 25 μg of anti-type A IgG and 350 μg of anti-type B IgG were equal to 1 IU. Neutralizing potency of F(ab′)2 was also analyzed as 30 and 350 μg/IU for type A and B toxins, respectively, showing that the neutralizing activity of the F(ab′)2 is close to that of IgGs. In this report, we successfully established the manufacturing process of equine botulinum toxin type A and B antitoxin and produced freeze-dried prototype products.

Quality of plasma and its fractionation

Human plasma (plasma for fractionation) is used as a source material for the manufacture of a large range of therapeutic products that are essential for the treatment of life‐threatening injuries as well as bleeding and immunological diseases. The industrial multistep process carried out to isolate the various therapeutic plasma proteins is known as ‘fractionation’. Typical plasma fractionation batches have a volume of 2000–4000 l obtained by pooling the individual donations from thousands of donors. The provision of safe plasma products relies upon the quality of the starting plasma and on a correct implementation of the fractionation processes, both under conditions meeting Good Manufacturing Practices (GMP) standards. The preparation of plasma for fractionation according to GMP is one key element in the quality and safety of medicinal plasma products. The quality specifications of plasma for fractionation should be precisely defined by the plasma fractionators and approved by the relevant regulatory authorities. Aspects of particular relevance for the control of infectious risks include (i) the selection criteria of blood/plasma donors, (ii) the methods used for the testing of blood/plasma donations, (iii) the epidemiological surveillance of the donor population, (iv) the adherence to GMP principles by blood establishments, and (v) the existence of a postdonation information system. Other quality aspects that may impact the range, quality, or recovery of fractionated products include the operational parameters used for plasma separation, rate of freezing and storage temperature. The industrial plasma fractionation process should be carried out under highly hygienic conditions in licenced facilities (plasma fractionation plants) operated in compliance with GMP. Manufacturing steps used to isolate the various fractions are implemented in a sequential and integrated manner. Crude fractions obtained by cryoprecipitation and cold ethanol precipitation steps are purified into individual therapeutic products most often by chromatographic methods. Validated dedicated treatments are implemented within the manufacturing process of each product to inactivate and/or remove enveloped and non‐enveloped viruses potentially present in the starting plasma pool. Recent experimental spiking studies also demonstrate that, several fractionation steps contribute to prion removal. When using current production technologies and under strict regulatory oversight, plasma products have reached a high level of quality and safety.

Investigations of prion and virus safety of a new liquid IVIG product

A highly purified, liquid, 10% immunoglobulin product stabilized with proline, referred to as IgPro10 has recently been developed. IgG was purified from human plasma by cold ethanol fractionation, octanoic acid precipitation and anion-exchange chromatography. The manufacturing process includes two distinctly different partitioning steps and virus filtration, which were also assessed for the removal of prions. Prion removal studies used different spike preparations (brain homogenate, microsomes, purified PrP sc) and three different detection methods (bioassay, Western blot, conformation-dependent immunoassay). All of the investigated production steps were shown to reduce significantly all different spike preparations, resulting in an overall reduction of >10 log 10. Moreover, the biochemical assays proved equally effective to the bioassay for the demonstration of prion elimination. Four of the manufacturing steps cover three different mechanisms of virus clearance. These are: i) virus inactivation; ii) virus filtration; and iii) partitioning. These mechanisms were assessed for their virus reduction capacity. Virus validation studies demonstrated overall reduction factors of >18 log 10 for enveloped and >7 log 10 for non-enveloped model viruses. In conclusion, the IgPro10 manufacturing process has a very high reduction potential for prions and for a wide variety of viruses resulting in a state-of-the-art product concerning safety towards known and emerging pathogens.

Macrophage Migration Inhibitory Factor Induces B Cell Survival by Activation of a CD74-CD44 Receptor Complex

Macrophage migration inhibitory factor (MIF) is an upstream activator of innate immunity that regulates subsequent adaptive responses. It was previously shown that in macrophages, MIF binds to a complex of CD74 and CD44, resulting in initiation of a signaling pathway. In the current study, we investigated the role of MIF in B cell survival. We show that in B lymphocytes, MIF initiates a signaling cascade that involves Syk and Akt, leading to NF-kappaB activation, proliferation, and survival in a CD74- and CD44-dependent manner. Thus, MIF regulates the adaptive immune response by maintaining the mature B cell population.

A new liquid intravenous immunoglobulin with three dedicated virus reduction steps: virus and prion reduction capacity.

A new 10% liquid human intravenous immunoglobulin (US trade name: Gammagard Liquid; European trade name: KIOVIG) manufactured by a process with three dedicated pathogen inactivation/removal steps (solvent/detergent treatment, 35-nm nanofiltration and low pH/elevated temperature incubation) was developed. The ability of the manufacturing process to inactivate/remove viruses and prions was investigated. Virus and prion removal capacities were assessed with down-scale spiking experiments, validated for equivalence to the large-scale process. Lipid-enveloped viruses were completely inactivated/removed by each of the three dedicated virus clearance steps, and for human immunodeficiency virus 1 (HIV-1) and pseudorabies virus (PRV), also by the upstream cold ethanol fractionation step. Relevant non-enveloped viruses [i.e. hepatitis A virus (HAV) and parvovirus B19 (B19V)] were effectively removed by nanofiltration and the cold ethanol fractionation step, and partial inactivation of non-enveloped viruses was achieved by low pH incubation. Overall log reduction factors were > 20.0 for HIV-1, > 18.1 for bovine viral diarrhoea virus, > 16.3 for West Nile virus, > 10.0 for influenza A virus subtype H5N1, > 21.8 for PRV, 12.0 for HAV, > 12.1 for encephalomyocarditis virus, 10.6 for B19V and 10.3 for mice minute virus. Prions (Western blot assay) were completely removed (> or = 3.2 mean log reduction) by a step of the cold ethanol fractionation process. Introducing three dedicated virus-clearance steps in the manufacturing process of immunoglobulins from human plasma provides high margins of safety.

The delivery and antinociceptive effects of morphine and its ester prodrugs from lipid emulsions

Long-acting analgesia is critical for patients suffering from long-acting pain. The purpose of this study was to develop lipid emulsions as parenteral drug delivery systems for morphine and its ester prodrugs. Morphine prodrugs with various alkyl chain lengths, including morphine propionate (MPR), morphine enanthate (MEN), and morphine decanoate (MDE), were synthesized. The prodrugs were stable against chemical hydrolysis in an aqueous solution, but were quickly hydrolyzed to the parent drug when exposed to esterase and plasma. Lipid emulsions were prepared using phosphatidylethanolamine (PE) as an emulsifier, while squalene was used as an inner oil phase. Drug release was found to be a function of the drug/prodrug lipophilicity, with a lower release rate for more-lipophilic drug/prodrugs. The inclusion of morphine and its prodrugs into lipid emulsions retarded their release. Lipid emulsions, which incorporated cholesterol, generally exhibited a drug/prodrug release comparable to that of emulsions without co-emulsifiers. Pluronic F68 (PF68) further slowed down the release of morphine and its prodrugs from the emulsions. The antinociceptive activity through the parenteral administration of these emulsions was examined using a cold ethanol tail-flick study. Compared with an aqueous solution, a prolonged analgesic duration was detected after application of the drug/prodrug emulsions. Incorporation of co-emulsifiers such as PF68 and cholesterol further increased the duration of action. The combination of prodrug strategy and lipid emulsions may be practically useful for improving analgesic therapy with morphine.

Myelin Basic Protein-primed T Cells Induce Neurotrophins in Glial Cells via α5β3 Integrin

Increasing the level of neurotrophins within the central nervous system may have therapeutic efficacy in patients with various neurological diseases. Earlier we have demonstrated that myelin basic protein (MBP)-primed T cells induce the expression of various proinflammatory molecules in glial cells via cell-to-cell contact. Here we describe that after Th2 polarization by gemfibrozil or other drugs, MBP-primed T cells induced the expression of neurotrophic molecules such as brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3), but not proinflammatory molecules in microglia and astroglia via cell-to-cell contact. MBP-primed Th2 cells expressed alpha5 and beta3 integrins and functional blocking antibodies against both alpha5 and beta3 integrins inhibited the ability of MBP-primed Th2 cells to induce glial neurotrophins. On the other hand, glial cells expressed PDGF-Rbeta and neutralization of this glial receptor abrogated the ability of Th2 cells to induce neurotrophins in glia. Activation of glial cAMP response element-binding protein (CREB) by MBP-primed Th2 cell contact and inhibition of contact-mediated expression of neurotrophins by antisense knockdown of glial CREB suggest that MBP-primed Th2 cell-glia contact induces the expression of neurotrophins through glial activation of CREB. Accordingly, blocking of either alpha5beta3 integrins on T cells or PDGF-Rbeta on glial cells impaired the ability of MBP-primed Th2 cells to induce glial activation of CREB. Furthermore, we demonstrate that these MBP-primed Th2 cells entered into the central nervous system and increased the expression of neurotrophins in vivo in the brain. This study illuminates the importance of alpha5beta3 and PDGF-Rbeta in guiding the novel neurotrophic property of neuroantigen-primed T cells via activation of CREB that may be of therapeutic importance in various neurological disorders.

Temperature Dependence of Solvation Dynamics of Probe Molecules in Methanol-Doped Ice and in Liquid Ethanol

We have studied the solvation statics and dynamics of coumarin 343 and a strong photoacid (pK* approximately 0.7) 2-naphthol-6, 8-disulfonate (2N68DS) in methanol-doped ice (1% molar concentration of methanol) and in cold liquid ethanol in the temperature range of 160-270 K. Both probe molecules show a relatively fast solvation dynamics in ice, ranging from a few tens of picoseconds at about 240 K to nanoseconds at about 160 K. At about 160 K in doped ice, we observe a sharp decrease of the dynamic Stokes shift of both coumarin 343 and 2N68DS. Its value is approximately only 200 cm-1 at approximately 160 K compared to about 1100 cm-1 at T >/= 200 K (at times longer than t > 10 ps). We find a good correlation between the inefficient and slow excited-state proton-transfer rate at low-temperature ice, T < 180 K, and the dramatic decrease of the solvation energy, as measured by the dynamic band shift, at these low temperatures. We find that the average solvation rate in ice is similar to its value in liquid ethanol at all given temperatures in the range of 200-250 K. The surprisingly fast solvation rate in ice is explained by the relatively large freedom of the water hydrogen rotation in ice Ih.

Disruption of the polyhydroxyalkanoate synthase gene inAeromonas hydrophilareduces its survival ability under stress conditions

Aeromonas hydrophila 4AK4 produces PHBHHx copolyesters consisting of 3-hydroxybutyrate and 3-hydroxyhexanoate. In this paper, the physiological functions of PHBHHx on the bacterial survival ability were systematically studied using a PHBHHx synthase PhaC-disrupted mutant termed A. hydrophila CQ4 and its wild type A. hydrophila 4AK4. It was found that PHBHHx synthesis in A. hydrophila 4AK4 provided improved resistance against environmental stress factors including heat and cold treatments, hydrogen peroxide, UV irradiation, ethanol and high osmotic pressure compared with its PHBHHx synthase negative mutant A. hydrophila CQ4. The above resistant abilities were further confirmed by the different expression levels of sigma factor sigma(S) encoded by rpoS via real-time PCR study. The results suggested that rpoS played an important role in the enhanced resistance of host strain to stresses conferred by PHBHHx.

Temperature-sensitive hydrogels composed of chitosan and hyaluronic acid as injectable carriers for drug delivery

Temperature-sensitive hydrogels composed of poly( N-isopropylacrylamide) (PNIPAAm) with chitosan (CPN) and chitosan + hyaluronic acid (CPNHA) were grafted in order to examine their physicochemical characteristics, in vitro drug release, and in vivo pharmacodynamics. The sol–gel transition behavior was investigated by UV/visible spectrophotometry, differential scanning calorimetry, and viscometry. A slight difference in the transition temperatures was observed among these polymer systems, with CPN and CPNHA exhibiting higher temperatures compared with PNIPAAm. A zeta potential determination revealed a positive charge for the CPN hydrogel, whereas no or only a negligible charge was observed for PNIPAAm and CPNHA. The entanglement of CPN hydrogels observed using scanning electronic microscopy showed the densest cross-linkage structure, followed by CPNHA and PNIPAAm. Both hydrophilic and lipophilic drugs, including nalbuphine, indomethacin, and the nalbuphine prodrug, were used as model drugs in an in vitro drug release experiment. All 3 hydrogels significantly prolonged drug release. The release rate of hydrophilic nalbuphine increased in the order CPN < CPNHA < PNIPAAm. The drug release of these hydrogels exhibited a trend opposite to that of lipophilic drugs. A cold ethanol tail-flick study was utilized in order to examine the antinociceptive activity of intravenous nalbuphine. CPN and CPNHA prolonged the analgesic duration of nalbuphine with no influence on the onset time. The loading of nalbuphine in the CPNHA hydrogel exhibited the longest analgesic duration (4 h) among the 3 hydrogels tested.

Partial purification and chemical characterization of a glycoprotein (putative hydrocolloid) emulsifier produced by a marine bacterium Antarctobacter

During screening for novel emulsifiers and surfactants, a marine alphaproteobacterium, Antarctobacter sp. TG22, was isolated and selected for its production of an extracellular emulsifying agent, AE22. This emulsifier was produced optimally in a low-nutrient seawater medium supplemented with glucose and was extractable by cold ethanol precipitation of the high-molecular-weight fraction (>100 kDa). Production of AE22 commenced towards the late exponential phase of growth, with maximum emulsifying activity detected after approximately 4 days of the cells entering the death phase. Chemical, chromatographic and nuclear magnetic resonance spectroscopic analysis confirmed AE22 to be a high-molecular-weight (>2,000 kDa) glycoprotein with high uronic acids content, thus denoting an apparent polyanionic structure. Functional characterization showed this polymer to compare well to xanthan gum and gum arabic as an emulsion-stabilizing agent for a range of different food oils. However, AE22 exhibited better stabilizing than emulsifying properties, which could be conferred by its viscosifying effect in solution or from certain chemical groups found on the polysaccharide or protein moieties of the polymer. This new high-molecular-weight glycoprotein exhibits interesting functional qualities that are comparable to other biopolymers of this type and shows particular promise as an emulsion-stabilizing agent in biotechnological applications.

Antibacterial properties of Uvaria chamae, Congronema latifolium, Garcinia kola, Vemonia amygdalina and Aframomium melegueta

The antimicrobial efficacy of cold and hot water and ethanol extracts of Garcinia kola, Congronema latifolium, Aframomium melegueta, Vemonia amygdalina and Uvaria chamae on Staphylococcus aureus, Streptococcus pyogenes, Escherichia coli, Salmonella typhi, Pseudomonas aeruginosa and Vibrio spp. were determined using well in agar diffusion technique. Cold and hot water extracts of G. kola and U. chamae moderately inhibited the growth of S. aureus and S. pyogenes with zone of inhibition of between 9 – 15 mm. V. amygdalina, G. kola and C. latifolium slightly inhibited S. pyogenes and E. coli with a zone of clearing of between 7 -13 mm. Cold or hot ethanol extracts of U. chamae, G. kola and V. amygdalina profoundly inhibited the growth of S. aureus, S. pyogenes, E. coli and S. typhi to about 13 to 21 mm. Also ethanol extract of C. latifolium inhibited the growth of S. aureus, S. pyogenes and E. coli with zone size between 13 to 20 mm. While P. aeruginosa was slightly inhibited by ethanol extracts of G. kola, A. melegueta and U. chamae. Soxhlet extracts of U. chamae, G. kola, V.Amygdalina and C. latifolium profoundly inhibited the growth of S. aureus, S. pyogenes, E. coli and S. typhi with zone of inhibition ranging from 13 – 22 mm. Vibro spp. were not inhibited by the cold and hot extract as well as soxhlet extracts of all the plants tested. The standard microorganisms, E. coli NCTC 10418 and S. aureus NCTC 6571, were moderately inhibited by the various test plant extracts with zones of inhibition ranging between 8 mm to 20 mm. This study reveals the antibacterial potentials of these plants.

Safety of IGIV therapy and infusion-related adverse events

Patients with primary immunodeficiency diseases have been treated with intravenous gammaglobulin (IGIV) for over 20 years. Gamma globulins were first introduced as a therapeutic modality in 1952 by Robert A. Good, who injected gamma globulins by the intramuscular (IM) route to treat patients with X-linked agammaglobulinemia. Subsequently, the clinical indications for gamma globulins therapy expanded from patients with Bruton's disease and other primary immune deficiency disorders to include a variety of autoimmune and inflammatory diseases. This expansion in clinical indications paralleled two critical steps in the development of gamma globulins preparations. First, proteolytic enzyme-treated immunoglobulins became available for intravenous (IV) administration; unlike earlier IM preparations, IV preparations could be administered in larger volumes, were less painful, rapidly achieved physiologic concentrations of serum IgG, and were similar to native immunoglobulins with respect to their distribution of IgG subclasses and their pharmacokinetic features. Newer, highly purified IGIVs also became available. Second, significant efforts have been made to enhance the viral safety of IGIVs. Manufacturers no longer depend on Cohn-Oncley or cold ethanol treatment as the only viral elimination step; rigorous donor screening, plasma testing by sensitive nucleic acid testing, and additional viral inactivation steps in the manufacturing processes for IGIVs have been implemented. In spite of the fact that this therapy has been life-saving in patients with primary immunodeficiency diseases, a number of adverse events have been reported as a result of its use. This paper will review some of the more frequent and important adverse events associated with the administration of IGIV, and discuss the mechanisms by which these reactions may occur. Our approach for replacement therapy with IGIV of patients with primary immunodeficiency diseases will be described, and some of the concerns related to the availability of commercial IGIV for this important patient population will be presented. Availability of IGIV product has been of particular concern to the clinical immunologists who take care of patients with primary immune deficiency disorders since there are no alternative therapies for these patients.