The development ofAzotobacter in the oat rhizosphere and its effect on the yield

ИccлeДOBAлocь paэBиTиe paэлиЧHьix Πitammb aэotoбaкtepa b pиэocфepe obca и иx bлияhиe ha ypoжa и. ΠepbohaЧaлЧhЧie Πitammьi cpabhиbaлиcь c t. h. iiaccaж иpobahhьimh шtammamи (te жe шtammьi, Aotopьie длиte иьho пobtopho bьipaщиba лиcь B pиэOCфepe OBca, pocшe гo B CTepилиэoBaHHoй Π). БaкTepиэaция ceMяB OBca BceMи шTaMMaMи aэOTOбaкTepa Bлиялa лa Ha кOлиЧecTBO aэOToбaкTepa B ΠOЧBe PиэOCъepьI, HO He Ha кOPHяX. Πaccaжи aэOTOбaкTepa Чepeэ pиэocфepy OBca B HaшиX ycлoBияX OΠьITa He yBeлиЧиBaли CΠOCOбHOCTи эTиX шTaMMOB paэBиBaTьCя бOлee yCилeHHO Ha кOPHяX или B ΠOЧBe PиэOCфePяI, ЧeM пePBOHaЧaлпHпIe шTaMMьI, — B OTлиЧиe OT дaHHьIX OпьITOB CO шTaMMaMи aэOTOбaкTepa, кOTOPьIe BьIPaщиBaлиCь C AOPHяMи PaCTeHий B CPeдe C aг;apoM (эиHOBьeBa 1950‐1954, BaHЧypa C COTP. 1959). БaкTepиэaпия BCeMи шTaMMaMи aэOTOбaкTepa цOBяIшaлa ypoжaй B CPaBHeHии C кOHTpOлeM.     

Der Einfluss einiger Substanzen auf die Bildung vergrösserter,starkwandiger Zellen beim PilzAspergillus niger

Пpи глyбиHHOй кyльTиBaции кpи бкa Aspergillus niger Ha кaчaлкe ΠepeMeHHo-BoзBpaTHOгO дBиж eHия B paCTBOpe MeлaccьI BcTpeчaлиcь гифьI c yBeл ичeHHьIMи TOлCTOCTeHHьIMи, OбьIчHO иHTepкaляpHO pacΠOлOжeHHьIMи клeTкaMи, кOTOpьIe бOльшe BCeгO HeΠOMиHaли XиaMидOCΠOpьI y CeMeЙcTBa Mucoraceae или клeTки HaзьIBaeMьIe Hülle-Zellen. B ΠpиcyT-cTBии cooTBeTcTBeHHьIX кOHцeHTpaций гидpOкcилaMиHa c aзoTHoкиcльIM aMMo-HиeM кoличecTBo yBeличeHHьIX клeToк yжe Ha BTopoй дeHь кyльTиBaции зHaчи-Teл ьHo BoзpacTaeT. MeHee эффeкTиBHOй oкaзaлacь кoMбиHaция гидpoкcил aMиHa, aзoTHoкиcлoгo HaTpия cepHoкиcлoгo aM-MoHи я. ЗaΠoздaлoe oбpaзoBaHиe бoль-шoгo к oличecTBa yBeличeHHьIX клeToк Haблюдaлocь B ΠpиcyTcTии HeкoTopьIX aMиHoкиcлoT, B ocoбeHHocTи глюTaMи-HoBoй киcлoTьI, a дaдee циcTeиHa, aлa-HиHa и TиpoзиHa. OбcyжлaюTcя Boз-MoжHocTи дeйcTBия гидpoкcилaMиHa c ocTaльHьIMи ΠpиMecяMи Ha oбpaзoBaHиe yBeличeHHьIX клeToк.     

Fluorescent antibody method Conjugation Of Fluoresceinisothiocyanate With Immune ?-Globulin

ИЗyчaл0C044A; Bлияниe НeкOTOpыX ФaкTopoB (_pH, OTHoшeниe КpacиTeля к бeлкy, КOнпeнTpaция КOMпoнeнTOB, пpиcyTcTBиe opraничecкиX pacTBopиTeлeй B peaгиpyюшeй cмecи, TeMпepaTypa, Bpeмя peaкции) Ha эффeкTиBн0CTъ кOнъюгaции γ-глoбyлинa c флyopecцeинизOTиoциaнaTOM. КoличecTBO КpacиTeля, CBязaннoгo c γ-глoбyлинOM, знaч иTeлънO пoBышaлocъ, кoгдa пoBышaлcя B пpeдeлax 7,0–10,0_pH peaгиpyющeй cмeCи, Toгдa кaк ocTaлъныe фaкTopы нe oкaзыBaли нa мeчeныe бeлки B иccлeдyeмыx пpeдeлax cyщecTBeннoгo Bлияния. БылO ycTaнOBлeнo, чTO cпeцифичecкaя флyopecцeнпия cBязaннoгo кpacиTeля зaмeTнo пoнижaлacъ c пOBышeниeм мeчeннOcTи бeлкOB, a пoэTOмy нaибoлee цeлecooбpaзным пpeдcTaBляeTCя иcпoлъзOBaниe для мeTOдa флyopecцeнTныx aнTиTeл-кOнъюгaTOB c BecoBым COOTнOшeниeм бeлкOB и кpacиTeля, paBным пpиблизиTeльнO 60.     

The effect of extracellular enzymes of wood-rotting fungi on wood

YKa3bIBaeMBIe B JIиTepaType IIpиeMbI биOJIOГиЧecKOЙ OбpaбOTKи ДpeBecиHbI иCnoJIb3yЮT ДeЙcTBиe pacTyЩиx ДpeBecHbIx rpибKOB. ЭTO — cЦocoб IIpocToЙ, HO, BO-IIepBbIx, MeДJIeHHbIЙ, a BO-BTOPBIX, He rapaHTиpyЮЩиЙ; paBHOMepHocTи ДeЙCTBиЯ. IIpиMeHЯЯ aKTиBHbIe npeπapaTbI; фepMeHTOB ДpeBecHbIx rpибKOB, MOжHO COKpaTиTb BeCb C0pOЦeCC ДO HeCKOJIbKиX ДecЯTKOB ЧacoB и IIpeДyπpeДиTb HepaBHOMepHoe pa3pyШneHиe CTpyKTypbI ДpeBeCиHbI.     

The influence of the soil structure and moisture content on the number of bacteria and the degree of nitrification

ИзгчалоAЬ одноврeмeнноe дeйствиe сBрCкСC@ы поGвKi и Aодeрeа влаBи на количe бактeрий и интeнсивноAтЬ нитрифика. ции в гумуA-карбонаBной почвe. Путeм просeиваниHя образцоCпочвы чeрeз сиBа были в044B;цeлeны слeдующиe фракции: I — агрeгaаты мeнЬшe 1 мм; II — агрeгаты размe@ами в 1—2; мм; III — аг@eгаты размeрами в 2—3 мм; IV — агтeгаBы размeрами в 3—4 мм; V — агрeгаBы размeрами в 4—5 мм. Образцы брали три @аза а год. Для опрeДeлeния количeства бактeрий полЬзовалисЬ прямым мeтодeм флуорeсцeнтной м0438;H043A;H0440;H043E;скопии по Strugger в модификации по Seifert. Пeрeд опрeдeлeниeм биогeнности образцы насыщли влагой до заранee установлeнного уровня и инкCбировали 48 час. при тeмпeратгрe 20° С Нитрификацию опрeдeлOли послe 14-днeвной ?нкгбации образFов, насKщeнных влвгой до опрeдeлeнного уровня и хранившихся в тeрмостатe пр? 20° А. Бы;ло установлeно, что количeство бактeрий зависит от содeржания влаги и 043F;очти нe зависит оB размeров агрeгатов (рис. 1). ИнтeнсивностЬ нитрификации зависит как от содeржаниO влаги, так ? оB размeров агр;eгатов. НаиболЬHая интeнсивностЬ наблюдаласЬ в слу чаe агрeгатов пe@вой фракции, т. e. при диамeтрах мeнЬHe 1 мм. С гвeличeниeм диамeтров агрeгатов интeнсивностЬ нитрификации постeпeнно поннжаласЬ (@ис. 2). Понижeниe интeнсивности нитрификации отличалосЬ опрeдeлCнной правилЬностЬю, инаосновании ee анализа мы пришли к выводу, что инBeнсивностЬ нитрификаFии прямо пропорFионадЬна вeдиGинe спeцифичeAкой повeрхносBи агрeгатов. ДоказатdдЬство зтого привeдeно на рис. З. Для протeрки того, как на интeнсивносBЬ нитрификации дeйствуeт радмeлЬчeниe агрeгатов, мы поставнди опыт, при котором апрeгаBы V фракции были размeлЬчeны до размeров I фракции. Послe измeлЬчeниO интeнсивностЬ ниBририкаFии знаGиBeлЬно повысиласЬ (рис. 4).     

Book reviews

Book reviewed in this article:
B asic B iotechnology . A S tudent's G uide (1987). Edited by P. Prave, U. Faust, W. Sittig & D.A. Sukatsch.
B acteria AS P lant P athogens (1987). By Eve Billing.
B iotechnology. A C omprehensive T reatise I n 8 V olumes .
C urrent T opics I n M icrobiology A nd I mmunology 136: T he M olecular B iology O f B acterial V irus S ystems (1988). Edited by G. Hobom & R. Rott.
G enetic B iochemistry : F rom G ene T o P rotein (1988). By J. Etienne-Decant.
I ntracellular B acteria : C urrent T opics I n M icrobiology A nd I mmunology N o.
P roceedings O f T he 8TH I nternational B iotechnology S ymposium P aris 1988 (1989). Edited by G. Durand, L. Bobichon & J. Florent. Vol.
B iological W aste T reatment (1989). Edited by A. Mizrahi.
I n V itro T echniques I n R esearch (1989). Edited by J.W. Payne.
B loodstream I nfections : L aboratory D etection A nd C linical C onsiderations (1989). By C.L. Strand & J.A. Shulman.
M etal -M icrobe I nteraction (1989). Edited by Robert K. Poole & Geoffrey M. Gadd.
B iochemistry O f A ntimicrobial A ction (1989). By T.J. Franklin & G.A. Snow.
T heory A nd A pplication O f M icrobial A ssay (1989). By W. Hewitt & S. Vincent.     

Book reviews

Book reviewed in this article:
E lectrochemical D etection T echniques I n T he A pplied B iosciences Volume 2 (1988). By G-A. Junter
P lant R esistance T o V iruses (1987). Edited by D. Evered & S. Harnett
M icrobial R econtamination I n D airy P rocessing –P roceedings O f T he I nternational C ongress.
D evelopments I n F ood M icrobiology –4 (1988). Edited by R.K. Robinson.
C ellulose H ydrolysis (1987). Edited by L. T. Fan, M.M. Gharpuray & Y.-H. Lee
I mmobilization O f C ells (1988). By C.R. Phillips & Y.C. Poon
C rystalline B acterial C ell S urface L ayers (1988). Edited by U.B. Sleytr, P. Messner, D. Pum & M. Sara     

Influence of pH, temperature, and urea molar flowrate on Arthrospira platensis fed-batch cultivation: a kinetic and thermodynamic approach

Arthrospira platensis was cultivated photoautotrophically at 6.0 klux light intensity in 5.0-L open tanks, using a mineral medium containing urea as nitrogen source. Fed-batch experiments were performed at constant flowrate. A central composite factorial design combined to response surface methodology (RSM) was utilized to determine the relationship between the selected response variables (cell concentration after 10 days, X(m), cell productivity, P(X), and nitrogen-to-cell conversion factor, Y(X/N)) and codified values of the independent variables (pH, temperature, T, and urea flowrate, K). By applying the quadratic regression analysis, the equations describing the behaviors of these responses as simultaneous functions of the selected independent variables were determined, and the conditions for X(m) and P(X) optimization were estimated (pH 9.5, T = 29 degrees C, and K = 0.551 mM/day). The experimental data obtained under these conditions (X(m) = 749 mg/L; P(X) = 69.9 mg/L.day) were very close to the estimated ones (X(m) = 721 mg/L; P(X) = 67.1 mg/L.day). Additional cultivations were carried out under the above best conditions of pH control and urea flowrate at variable temperature. Consistently with the results of RSM, the best growth temperature was 29 degrees C. The maximum specific growth rates at different temperatures were used to estimate the thermodynamic parameters of growth (DeltaH* = 59.3 kJ/mol; DeltaS* = -0.147 kJ/mol.K; DeltaG* = 103 kJ/mol) and its thermal inactivation (DeltaH(D) (o) = 72.0 kJ/mol; DeltaS(D) (o) = 0.144 kJ/mol.K; DeltaG(D) (o) = 29.1 kJ/mol).     

Structures and electron affinities of triatomic molecules consisting of Al, P and X (X = B, Al, Ga; C, Si, Ge; N, P, As; O, S and Se)

The structures and electronic properties of the triatomic molecules containing Al, P, X atoms (X = B, Al, Ga; C, Si, Ge; N, P, As; O, S and Se) and their anions are investigated at the B3LYP/cc-PVTZ and the B3LYP/aug-cc-PVTZ levels. The results show that the most stable structures of the anions are AlXP⁻ (X = B, C, N) and PAlX⁻ (X = S, Se), while for the neutral molecules, the most stable structures are PXAl (X = C, N and O). The order of the VDEs of the anions molecules and the AEAs of the neutral species are C < N < O < Si ≈ Ge < P ≈ As < Al = Ga < B < S ≈ Se and C < O < N < Si ≈ Ge < P ≈ As < B < Al ≈ Ga < S ≈ Se, respectively.     

Nuclear thyroxine and triiodothyronine binding in mononuclear cells in dependence of age

Nuclear binding of thyroxine (T4) and triiodothyronine (T3) in mononuclear blood cells was investigated in 12 young (age 16-30 years) healthy subjects (group A), in 12 middle-aged (age 31-60 years) healthy subjects (group B) and in 12 elderly (61-90 years) healthy subjects. Serum free T3 was depressed in group C as compared to the younger age groups, whereas serum free T4 and TSH did not differ between the groups. Maximal specific nuclear binding capacity for both T4 and T3 decreased with increasing age, T4 group A: 1.2 fmol T4/100 micrograms DNA, group B: 1.2 fmol T4/100 micrograms DNA, group C: 0.7 fmol T4/100 micrograms DNA; T3 group A: 1.7 fmol T3/100 micrograms DNA, group B: 1.0 fmol T3/100 micrograms DNA, group C: 0.9 fmol T3/100 micrograms DNA. The equilibrium association constant (Ka) for T4 increased with age, group A: Ka = 3.3 X 10(9) l/mol, group B: Ka = 3.2 X 10(9) l/mol, group C: Ka = 6.4 X 10(9) l/mol, whereas Ka for nuclear binding of T3 decreased with age group A: Ka = 3.9 X 10(9) l/mol, group B: Ka = 5.9 X 10(9) l/mol, group C: Ka = 1.8 X 10(9) l/mol. We conclude that, whereas the opposite variations of nuclear capacity and binding affinity for T4 tend to preserve the nuclear T4 concentration, the nuclear T3 concentration definitely decreases with age. The unaltered serum levels of TSH suggest that the decrease of both serum levels of free T3 and the nuclear T3 concentration might represent physiologically changes in old age.     

Structures of the O1B and O1C lipopolysaccharide antigens of Escherichia coli.

The O-specific moieties of the O1B antigen (lipopolysaccharide) from Escherichia coli O1B:K1 and the O1C antigen from E. coli O1C:K- both consist of L-rhamnose, D-galactose, N-acetyl-D-glucosamine, and N-acetyl-D-mannosamine in a molar ratio of 2:1:1:1. By using fragmentation procedures, methylation analysis, and one- and two-dimensional nuclear magnetic resonance spectroscopy, the structures of these polysaccharides were found to be [formula: see text] In the O1B polysaccharide X is 2, and in the O1C polysaccharide X is 3. With the recently published structure of the O1A polysaccharides (B. Jann, A. S. Shashkov, D. S. Gupta, S. M. Panasenko, and K. Jann, Carbohydr. Polym. 18:51-57 1992), three related O1 antigens are now known. Their common (O1-specific) epitope is suggested to be the side-chain N-acetyl-D-mannosamine residue.     

The Effect of Bovine IFN-α on the Immune Response in Guinea Pigs Vaccinatedwith DNA Vaccine of Foot-and-Mouth Disease Virus

Foot-and-mouth disease virus (FMDV) belongs to thegenus Aphthovirus of the family Picornavidae. The FMDVgenome is a copy of positive-sense, single-stranded RNA,which contains one large open reading frame (ORF). TheORF is translated into a polypeptide, which undergoesautoproteolytic cleavage to produce the structural and non-structural proteins and ultimately forms mature viral pro-teins [1,2]. FMD is caused by the FMDV, which is a highly conta-gious vesicular disease of cloven-hoofe…     

Selectivity of hemoregulatory peptide (HP5b) action in culture

A synthetic analog of a hemoregulatory peptide associated with mature human granulocytes (HP5b) has been investigated for inhibitory effects on various cell types in culture as compared to inhibitory action on mouse and human myelopoietic colonies (CFU-gm), which occurs from 1 X 10(-13) to 1 X 10(-6) M in vitro. This includes colony formation by lymphoid T and B cells in capillary cultures, as well as mitogen activation of T, B and NK cells. At higher concentrations, i.e., above 1 X 10(-7) M, an inhibitory effect was found on colony formation. Neither the production of interleukin (IL) 3 by mitogen-activated T cells, nor the proliferation of the IL-3-dependent L/B cell line were affected by the peptide up to 1 X 10(-5) M. A slight inhibitory effect was found above 1 X 10(-9) M on mouse 3T3 fibroblasts. A series of malignant cell lines was also tested. No effect was seen between 1 X 10(-11) and 1 X 10(-7) M on human mammary carcinoma cells in culture. On Ehrlich ascites mouse mammary carcinoma cells a 30% inhibition was seen at 10(-6) M. On a human glioblastoma cell line (GaMg) no effect was seen, and on a rat glioma cell line (BT5C) an inhibitory effect was seen at 1 X 10(-7) M and above. No significant inhibition of cell growth was seen on SC1 mouse lymphoma cells from 1 X 10(-9) to 1 X 10(-5) M during 7 days of culture. The investigated normal and malignant cell types in culture were thus not inhibited in very low concentrations which act on CFU-gm. However, a variable inhibitory effect was found at higher concentrations where the inhibition of myelopoiesis was maximal and at concentrations where the inhibition is released. The hemoregulatory peptide thus seems to be a concentration-dependent selective inhibitor of myelopoiesis. The finding that various malignant cells do not respond at lower concentrations supports the possibility of using the peptide as a protector of normal cells during cancer chemotherapy.     

Comparison of the heme structures of horseradish peroxidase compounds X and II by resonance Raman spectroscopy

Horseradish peroxidase will catalyze the chlorination of certain substrates by sodium chlorite through an intermediate known as compound X. A chlorite-derived chlorine atom is known to be retained by compound X and has been proposed to be located at the heme active site. Although several heme structures have been proposed for compound X, including an Fe(IV)-OCl group, preliminary data previously reported by our laboratory suggested that compound X contained a heme Fe(IV) = O group, based on the similarity of a compound X resonance Raman band at 788 cm-1 to resonance Raman Fe(IV) = O stretching vibrations recently identified for horseradish peroxidase compound II and ferryl myoglobin. Isotopic studies now confirm that the 788 cm-1 resonance Raman band of compound X is, in fact, due to a heme Fe(IV) = O group, with the oxygen atom derived from chlorite. The Fe(IV) = O frequency of compound X, of horseradish peroxidase isoenzymes B and C, undergoes a pH-induced frequency shift, with behavior which appears to be the same as that previously reported for compound II, formed from the same isoenzymes. These observations strongly suggest that compounds II and X have very similar, if not identical, heme structures. The chlorine atom thus appears not to be heme-bound and may rather be located on an amino acid residue. The studies on compound X reported here were done in a pH region above pH 8, where compound X is moderately stable. The present results do not necessarily apply to compound X below pH 8.     

Synthesis and enzymatic characterization of P1-thio-P2-oxo trideoxynucleoside diphosphates having AZT, FdU, or dT at the 3'-position

Model compounds for oligonucleotide-prodrugs, P1-thio-P2-oxo-trideoxyribonucleoside diphosphates: d[G(s)C(o)X] and d[T(s)A(o)X] (X = AZT, FdU or dT) have been prepared, and their hydrolyses by snake venom phosphodiesterase and nuclease S1 are described.     

Specific antigen-binding and antibody-secreting lymphocytes associated with the erythrocyte autoantibody responses of NZB and genetically unrelated mice.

The receptor characteristics as well as incidence of antigen-binding lymphocytes (ABL) or B and T cell classes with membrane receptors specific for the exposed (X) and cryptic (HB) murine erythrocyte autoantigens were examined in NZB and nine control strains of mice. Whereas only NZB and NZB hybrid mice synthesize anti-X autoantibody pathogenetically implicated in the genetically determined autoimmune hemolytic anemia, the NZB as well as control strains synthesize the ubiquitous anti-HB anti-erythrocyte autoantibody. By utilizing immunocytoadherence assays, maximum numbers of specific ABL of both B and T lymphocyte classes were optimally demonstrated at erythrocyte:lymphocyte ratios of 20:1 and after lymphocyte fixation at 56 degrees C for 20 min. Surface membrane receptor specificity was established by inhibition with semi-purified soluble X or HB autoantigen. Inhibition of immunocyto-adherence with class specific antisera to mouse immuno-globulins demonstrated that the receptors on both B and T cells were of IgM class. Specific receptors regenerated in vitro after trypsinization which excluded the role of cytophilic antibody in the immunocytoadherence reactions. B lymphocyte ABL reactive with the X autoantigen were demonstrable in NZB, NZB hybrid, and control mice. Only in NZB and NZB hybrid mice, strains that uniformly synthesize anti-X autoantibody, were X ABL of T lymphocyte class demonstrated. The presence and incidence of T lymphocyte X ABL is compatible with the expression of a single dominant gene carried by the NAB strain. The incidence of B lymphocyte X ABL increased with age, suggesting proliferation of this cell population. HB ABL of both B and T lymphocyte classes were observed in all strains, concordant with the ubiquitous presence of humoral anti-HB autoantibodies. Differentiation of precursor B cells are evaluated by PFC assay of cells secreting specific autoantibodies. Anti-X PFC were observed only in NZB and NZB hybrid mice; and the observed frequency suggested that less than 3.5% of the specific ABL were differentiated for the secretion of anti-X autoantibody. Anti-HB PFC were observed in all strains and represented as high as 11.8% of specific ABL. Genetic determination of the anti-X anti-erythrocyte autoantibody response does not prescribe the presence of precursors of the antibody-forming cell, but rather appears to influence regulation of the differentiation of these cells. These data suggest that circumvention of immunologic tolerance to this specific erythrocyte autoantigen may occur at the level of the T lymphocyte; or alternatively, that T lymphocytes as well as B lymphocytes, are induced to proliferate and differentiate in the NZB strain.     

A kinetic description of ligand binding to sperm whale myoglobin

Nanosecond recombination time courses were measured by photolyzing O2, NO, CO, methyl, ethyl, n-propyl, n-butyl, and tert-butyl isocyanide complexes of sperm whale myoglobin with a 30-ns laser pulse at pH 7, 20 degrees C. Absorbance was measured both during and after the excitation pulse and as a function of laser light intensity. The results were analyzed quantitatively in terms of a three-step reaction scheme, MbX in equilibrium B in equilibrium C in equilibrium Mb + X, where Mb is myoglobin, B represents a geminate state in which the ligand is present in the distal pocket but not covalently bound to the iron atom, and C, a state in which the ligand is still embedded in the protein but further away from the heme group. The fitted rate parameters were required to be consistent with the observed overall quantum yield, Q, which had been measured independently using much longer (approximately 0.5 ms) xenon flash pulses. Three major conclusions were derived from these analyses. First, the overall quantum yield of the ligand complex is determined primarily by the competition between the rate of iron-ligand bond formation from the initial photoproduct, kB----MbX, and the rate of migration away from state B, kB----C. For example, kB----C approximately equal to 30-100 microseconds-1 for all three gaseous ligands, whereas both Q and kB----MbX vary over 3 orders of magnitude (i.e. NO, Q = 0.001, kB----MbX approximately equal to 16,000 microseconds-1; O2, Q = 0.1, kB----MbX approximately equal to 500 microseconds-1; CO, Q = 1.0, kB----MbX approximately equal to 2 microseconds-1). Second, for NO, O2, and the isonitriles, the rate-limiting step in the overall association reaction starting from ligand in solution is the formation of state B. The rate constant for this process varies from 2 X 10(7) M-1 s-1 for the gaseous ligands to 0.02-1.4 X 10(5) M-1 s-1 for the isonitriles. In contrast, the B to MbX transition is limiting for CO binding. Third, for all the ligands except CO, the overall rate of dissociation is limited significantly both by the rate of thermal bond disruption, kMbX----B, and the competition between geminate recombination and migration away from the distal pocket (i.e. kB----C/(kB----MbX + kB----C]. In the case of CO, the rate of bond disruption is equal to the observed dissociation rate constant.     

Ligand binding equilibrium and kinetic measurements on the dimeric myoglobin of Busycon canaliculatum and the comparative ligand binding of diverse non-cooperative heme proteins

The rate constants and delta H degrees for the non-cooperative dimeric Busycon myoglobin are: oxygen, k' = 4.75 X 10(7) M-1 sec-1, k = 71 sec-1, and CO, l'= 3.46 X 10(5) M-1 sec-1, l = 0.0052 sec-1 at 20 degrees C, pH 7, delta H degrees = -3 kcal/mol for O2 and CO.2. Log-log plots of k vs K for oxygen and of l' vs L for CO binding for numerous non-cooperative hemoglobins and myoglobins point to a large steric influence of the protein on heme ligation reactions. Many of the proteins behave as "R" state for one ligand, but "T" for the other.     

Growth plate compressions and altered hematopoiesis in collagen X null mice

A variable skeleto-hematopoietic phenotype was observed in collagen X null mice which mirrored the defects in transgenic (Tg) mice with dominant interference collagen X mutations (Jacenko, O., P. LuValle, and B.R. Olsen. 1993. Nature. 365:56-61). Specifically, perinatal lethality was seen in approximately 10.8% of null mutants at week three after birth, and in another subset by 12 wk. In perinatal lethal mutants, growth plates were compressed, trabecular bone reduced, and hematopoietic aplasia and erythrocyte-filled vascular sinusoids were apparent in marrows. Lymphatic organs, reduced to approximately 80% that of controls, displayed altered architecture and lymphocyte content. In thymuses, a paucity of cortical CD3(+)/CD4(+)/CD8(+) lymphocytes was consistent with the marrow's inability to replenish maturing T cells. In spleens, an unaltered T cell distribution was coupled with diffuse staining for IgD(+)/B220(+) B cells, whose reduction was prominent in poorly organized lymphatic nodules. Disorderly arrays of splenic macrophages surrounding periarteriolar lymphatic sheaths and a red pulp depletion further complemented the Tg perinatal lethal phenotype. Moreover, subtle growth plate compressions and hematopoietic changes were seen in all null mice. Data from Tg and null mice implicate the disruption of collagen X function in the observed skeleto-hematopoietic defects, and suggest that hypertrophic cartilage and endochondral skeletogenesis may contribute to the marrow microenvironment prerequisite for blood cell differentiation.     

HIV-1 Clade B Tat, but Not Clade C Tat, Increases X4 HIV-1 Entry into Resting but Not Activated CD4+ T Cells

CXCR4-using human immunodeficiency virus, type 1 (HIV-1) variants emerge late in the course of infection in >40% of individuals infected with clade B HIV-1 but are described less commonly with clade C isolates. Tat is secreted by HIV-1-infected cells where it acts on both uninfected bystander cells and infected cells. In this study, we show that clade B Tat, but not clade C Tat, increases CXCR4 surface expression on resting CD4⁺ T cells through a CCR2b-dependent mechanism that does not involve de novo protein synthesis. The expression of plectin, a cytolinker protein that plays an important role as a scaffolding platform for proteins involved in cellular signaling including CXCR4 signaling and trafficking, was found to be significantly increased following B Tat but not C Tat treatment. Knockdown of plectin using RNA interference showed that plectin is essential for the B Tat-induced translocation of CXCR4 to the surface of resting CD4⁺ T cells. The increased surface CXCR4 expression following B Tat treatment led to increased function of CXCR4 including increased chemoattraction toward CXCR4-using-gp120. Moreover, increased CXCR4 surface expression rendered resting CD4⁺ T cells more permissive to X4 but not R5 HIV-1 infection. However, neither B Tat nor C Tat was able to up-regulate surface expression of CXCR4 on activated CD4⁺ T cells, and both proteins inhibited the infection of activated CD4⁺ T cells with X4 but not R5 HIV-1. Thus, B Tat, but not C Tat, has the capacity to render resting, but not activated, CD4⁺ T cells more susceptible to X4 HIV-1 infection.