Biomedical subjects

A Porta

Publications and source records attributed to A Porta.

At least 37 records · Page 2Linked to original sources

PubMed2001

Contrasting effects of phentolamine and nitroprusside on neural and cardiovascular variability.

The relative contributions of a central neural oscillator and of the delay in alpha-adrenergic transmission within the baroreflex loop in the predominance of low-frequency (LF) cardiovascular variability during sympathetic activation in humans are unclear. We measured R-R interval (RR), muscle sympathetic nerve activity (MSNA), blood pressure (BP), and their variability in 10 normal subjects during sympathetic activation achieved by BP lowering with sodium nitroprusside (SNP) and alpha-adrenergic blockade using phentolamine. SNP and phentolamine induced comparable reductions in BP (P > 0.25). Despite tachycardia and sympathetic activation with both SNP and phentolamine, LF variability in RR, MSNA, and BP increased during SNP and decreased during phentolamine (SNP: RR +20 +/- 6%, MSNA +3 +/- 5%, systolic BP +9 +/- 6%, diastolic BP +7 +/- 5%; phentolamine: RR -2 +/- 7%, MSNA -34 +/- 6%, systolic BP -16 +/- 8%, diastolic BP -13 +/- 4%, P < 0.05 except systolic BP, where P = 0.09). Thus LF variability is reduced when sympathetic activation is induced by alpha-adrenergic blockade. This suggests that alpha-adrenergic transmission within the baroreflex loop may contribute importantly to the predominance of LF cardiovascular variability associated with sympathetic excitation in humans.

Adrenergic alpha-Antagonists↗

PubMed2001

[Spectral analysis of muscle sympathetic nerve activity in man].

Recent applications of frequency domain analysis to the variability of muscle sympathetic nerve activity (MSNA) have improved the comprehension of the relationship between cardiovascular oscillations and the autonomic nervous system. It has been observed that spectral analysis of MSNA is characterized by two major oscillatory components at low (LF) and high (HF) frequencies, similar to those detectable in the variability of cardiovascular signals. Pharmacological and non-pharmacological studies have shown that, at least within the physiological range, the two MSNA rhythms show a reciprocal behavior, similar to that already observed for cardiovascular oscillations. The oscillatory pattern of MSNA provides non-redundant but complementary information with respect to the time domain measures of MSNA (burst rate and amplitude) since it has been shown that completely different spectral profiles may be derived from various MSNA recordings. On the other hand, the latter are instead comparable in terms of mass activity. Due to the intrinsic characteristics of the signal, which represents the direct outflow of the central neural structures of the cardiovascular autonomic nervous system, this approach can be considered as a unique window over the central organization of excitatory and inhibitory neural mechanisms responsible for the genesis and the regulation of cardiovascular oscillations.

Electrophysiology↗

PubMed2001

Discrimination of atrial rhythms by linear and non-linear methods.

The discrimination among atrial rhythms is obtained through the analysis of linear and non-linear dynamics of atria electrograms and local atrial period (LAP) series. Linear and non-linear degree of coupling between atrial electrograms have been assessed using cross-correlation (CCI) and synchronization indexes. Dynamics of LAP series were investigated using level of predictability (LP) and regularity indexes. Atrial fibrillation (AF) episodes, classified according to Wells' criteria, were investigated. We found that metrics obtained from LAP series provide the best performance in the classification of AF rhythms. The LP index misclassified only 12 AF episodes over 87. Synchronization index was the most performing metric in the discrimination between organized and not-organized fibrillation: detection of organized rhythm; sensitivity (SE) > 95%; positive predictability (+P) > 95%. LP (SE: 89%; +P: 89%) and CCI (SE: 87%; +P: 83%) provide slightly lower performances.

Atrial Fibrillation↗

PubMed2000

Linear and non-linear 24 h heart rate variability in chronic heart failure.

It has recently been demonstrated that SDNN of heart rate variability (HRV) is a useful independent prognostic tool in chronic heart failure (CHF). The purpose of the present study was to evaluate if spectral and non-linear analysis of 24-h HRV, considered markers of autonomic cardiac modulation, contain independent prognostic information in CHF patients. Twenty normal subjects and thirty consecutive outpatients with clinically stable CHF were studied for 2 years. Periods of 300 R-R intervals were analyzed from Holter recordings. The power spectral analysis, the slope of the linear relationship between log-power versus log-frequency (1/f), and the complexity content (using corrected conditional entropy; CCE) of the R-R series were calculated. The normalized power of the low frequency spectral component (LF) and the 1/f slope were significantly lower in patients compared to controls (respectively 30.1 +/- 3.0 vs. 48.6 +/- 3.4 and -1.27 +/- 0.04 vs. -1.08 +/- 0.05; P < 0.05). Moreover, the patients who died during the study presented a reduced LF (20.9 +/- 4.1 vs. 35.5 +/- 3.5 nu; P < 0.05) and a steeper 1/f slope (-1.40 +/- 0.09 vs. -1.21 +/- 0.04 nuts, P < 0.05) compared to survivors. These results remained significant in a logistic model including heart rate and SDNN. The information content present in spectral and non-linear analysis of HRV in CHF patients has prognostic relevance independently from the time domain measures of HRV. In particular, the reduction of LF power seems the best indicator among those considered.

Aged↗

PubMed2000

Heart rate variability patterns before ventricular tachycardia onset in patients with an implantable cardioverter defibrillator. Participating Investigators of ICD-HRV Italian Study Group.

Time- and frequency-domain analysis of heart rate variability (HRV) has been proven effective in describing alteration of autonomic control mechanisms and in identifying patients with increased cardiac and arrhythmic mortality. Patients with implantable cardioverter defibrillators offer the opportunity to evaluate HRV patterns before ventricular tachycardia (VT) and under control conditions. We therefore analyzed time- and frequency-domain parameters of short-term HRV and power-law behavior of RR interval time series at rest, at 15 to 30 minutes, and immediately before VT. In comparison to control conditions, lower values of mean cycle length duration and total power were observed before VT. Spectral analysis indicated that the low- to high-frequency ratio was significantly higher (5.5 +/- 0.6 vs 2.8 +/- 0.3) immediately before VT than during rest. Both findings were consistent with the shift of sympathovagal balance toward sympathetic predominance and reduced vagal tone. Before VT, a more negative value of the scaling exponent beta of the power-frequency relation (-1.57 +/- 0.04 vs -1.33 +/- 0.04) also confirmed the presence of an altered HRV pattern in comparison to controls. Thus, both abnormal autonomic modulation and dynamic patterns of HRV seem to characterize the minutes before arrhythmia onset in these patients.

Aged↗

PubMed2000

Heart rate variability is encoded in the spontaneous discharge of thalamic somatosensory neurones in cat.

1. We studied the spontaneous discharge variability of thalamocortical somatosensory neurones in the awake cat in order to disclose its possible information content. The presence of slow (0.09-1.39 Hz) regular fluctuations in the discharge rate of these cells during the waking state has been previously reported. Oscillations in a similar frequency range are known to characterize the activity of central and peripheral neurones pertaining to the autonomic nervous system and the variability of heart period (RR interval variability). 2. A surrogate data test, performed on our database, confirmed the presence of slow (0.05-1 Hz) non-random fluctuations in firing rate. 3. Linear regression detected the presence of an inverse relationship between the values of RR interval and the concurrent levels of neural discharge. 4. Frequency domain analysis indicated that a significant coupling between the two variability signals preferentially occurred in two frequency bands: in the frequency of the respiratory sinus arrhythmia and in correspondence with a slower rhythm (0.07-0.3 Hz), the two signals being in phase opposition in most of the cases. 5. Coherent fluctuations could also be observed when epochs of evoked activity were analysed, while coupling between the two variability signals appeared to be disrupted after sleep onset. 6. We conclude that RR interval variability, an internally generated dynamic related to basic visceral regulation, is encoded in the discharge of single somatosensory thalamocortical neurones during wakefulness. A possible interaction with the transmission of somatosensory information has to be evaluated.

Activity Cycles↗

PubMed2000

Oscillatory patterns in sympathetic neural discharge and cardiovascular variables during orthostatic stimulus.

BACKGROUND: We tested the hypothesis that a common oscillatory pattern might characterize the rhythmic discharge of muscle sympathetic nerve activity (MSNA) and the spontaneous variability of heart rate and systolic arterial pressure (SAP) during a physiological increase of sympathetic activity induced by the head-up tilt maneuver. METHODS AND RESULTS: Ten healthy subjects underwent continuous recordings of ECG, intra-arterial pressure, respiratory activity, central venous pressure, and MSNA, both in the recumbent position and during 75 degrees head-up tilt. Venous samplings for catecholamine assessment were obtained at rest and during the fifth minute of tilt. Spectrum and cross-spectrum analyses of R-R interval, SAP, and MSNA variabilities and of respiratory activity provided the low (LF, 0.1 Hz) and high frequency (HF, 0.27 Hz) rhythmic components of each signal and assessed their linear relationships. Compared with the recumbent position, tilt reduced central venous pressure, but blood pressure was unchanged. Heart rate, MSNA, and plasma epinephrine and norepinephrine levels increased, suggesting a marked enhancement of overall sympathetic activity. During tilt, LF(MSNA) increased compared with the level in the supine position; this mirrored similar changes observed in the LF components of R-R interval and SAP variabilities. The increase of LF(MSNA) was proportional to the amount of the sympathetic discharge. The coupling between LF components of MSNA and R-R interval and SAP variabilities was enhanced during tilt compared with rest. CONCLUSIONS: During the sympathetic activation induced by tilt, a similar oscillatory pattern based on an increased LF rhythmicity characterized the spontaneous variability of neural sympathetic discharge, R-R interval, and arterial pressure.

Adult↗

PubMed2000

Information domain analysis of cardiovascular variability signals: evaluation of regularity, synchronisation and co-ordination.

A unifying general approach to measure regularity, synchronisation and co-ordination is proposed. This approach is based on conditional entropy and is specifically designed to deal with a small amount of data (a few hundred samples). Quantitative and reliable indexes of regularity, synchronisation and co-ordination (ranging from 0 to 1) are derived in a domain (i.e. the information domain) different from time and frequency domains. The method is applied to evaluate regularity, synchronisation and co-ordination among cardiovascular beat-to-beat variability signals during sympathetic activation induced by head-up tilt (T), during the perturbing action produced by controlled respiration at 10, 15 and 20 breaths/min (CR10, CR15 and CR20), and after peripheral muscarinic blockade provoked by the administration of low and high doses of atropine (LD and HD). It is found that: (1) regularity of the RR interval series is around 0.209; (2) this increases during T, CR10 and HD; (3) the systolic arterial pressure (SAP) series is more regular (0.406) and its regularity is not affected by the specified experimental conditions; (4) the muscle sympathetic (MS) series is a complex signal (0.093) and its regularity is not influenced by HD and LD; (5) the RR interval and SAP series are significantly, though weakly, synchronised (0.093) and their coupling increases during T, CR10 and CR15; (6) the RR interval and respiration are coupled (0.152) and their coupling increases during CR10; (7) SAP and respiration are significantly synchronised (0.108) and synchronisation increases during CR10; (8) MS and respiration are uncoupled and become coupled (0.119) after HD; (9) the RR interval, SAP and respiration are significantly co-ordinated (0.118) and co-ordination increases during CR10 and CR15; (10) during HD the co-ordination among SAP, MS and the respiratory signal is larger than that among the RR interval, SAP, MS and the respiratory signal, thus indicating that the RR interval contributes towards reducing co-ordination.

Atropine↗

PubMed2000

Absence of sympathetic overactivity in Afro-Caribbean hypertensive subjects studied by heart rate variability.

Black hypertensives present a greater prevalence of left ventricular hypertrophy and an increased mortality compared to white hypertensives. Differences in sympathetic activity might contribute to explain these racial differences in hypertension. Nevertheless, previous laboratory studies did not show any increase of sympathetic activity direct to the heart in black subjects. The aim of the present study was to investigate the cardiac sympatho-vagal balance in black and white hypertensives analysing heart rate variability, during the entire 24 h. We analysed Holter recordings of 52 essential hypertensive patients, who had never received antihypertensive treatment, 26 of whom were black and 26 were white. Consecutive series of 300 beats, with 150 beats overlapped (approximately 600 series/day), were considered for the analysis in time and frequency domain. The mean 24-h value of the power of the low frequency spectral component (0.04-0.15 Hz), expressed in normalised units, ie a marker of sympathetic modulation, was significantly lower in the group of black patients compared to whites (respectively 40.0 +/- 2.1 vs 53.6 +/- 3.6 nu, P < 0.01). Similar results were observed for the LF/HF ratio, an index of the sympatho-vagal balance (respectively 4.11 +/- 0.58 vs 5.98 +/- 0.79; P < 0.05). In a multiple linear regression analysis, considering diastolic blood pressure, left ventricular mass index, race and age as independent variables, only race (P < 0.002) and age (P < 0.01) could independently predict the normalised low frequency power or the LF/HF ratio, as dependent variables. The results of this study suggest some blunting of the cardiac sympathetic neural modulation in black hypertensives compared to white hypertensives, during the entire 24 h.

Adult↗

PubMed2000

Host response and Histoplasma capsulatum/macrophage molecular interactions.

Histoplasma capsulatum is the etiological agent of histoplasmosis, a chronic respiratory infection that is generally asymptomatic in healthy individuals, but severe or fatal in patients who are immunosuppressed or otherwise debilitated. H. capsulatum is found as a mould in soil and becomes a pathogenic yeast in the mammalian host. The first line of defense that H. capsulatum faces during host invasion is the attack of polymorphonuclear neutrophils and resident macrophages. In animal models, once phagocytosed, H. capsulatum is not killed by fusion of the phago-lysosomes, instead it multiplies within non-activated macrophages and destroys them. Upon induction of cell-mediated immunity, cytokines activate macrophages and destroy the yeast cells. Some aspects of the fungus-macrophage interaction have been elucidated, and it is clear that some of the mechanisms by which H. capsulatum escapes the lethal effects of this very hostile environment, involve the regulation of specific genes. Recently, using the differential display reverse transcriptase polymerase chain reaction technique, a number of H. capsulatum genes that are induced after the yeasts are ingested by macrophages have been identified. However, the mechanisms that underlie the capacity of H. capsulatum to adapt to the new environmental conditions present in macrophages remain to be clarified.

Animals↗

PubMed2000

Evaluation of respiratory influences on left ventricular function parameters extracted from echocardiographic acoustic quantification.

This study was designed to assess, using the echocardiographic acoustic quantification technique, the influence of respiration on left ventricular (LV) function and its modifications connected with the ageing process, quantifying in a non-invasive way the respiratory contribution to the LV volume variability. An automated algorithm is applied to extract the beat-to-beat measurements of LV function parameters from the LV volume signal, obtained from recordings lasting a few minutes. Mean values, amount of variability and spectral content were studied in a population of 17 normal young (mean age 25 +/- 1 years) and 12 normal old (mean age 64 +/- 2 years) subjects. Mean values of the beat-to-beat measurements of LV function parameters were able to point out alterations connected with the ageing process in peak filling rate, peak atrial filling rate and peak ejection rate. Spectral analysis, applied to the extracted variability series, displayed a predominance of the high-frequency (HF) component corresponding to respiration in all LV function parameters; moreover, age related changes of HF variability were observed in peak ejection rate. The HF power spectrum component of beat to beat series extracted from the LV signal can provide a non-invasive assessment of the fluctuations in ventricular parameters associated with respiration.

Adult↗

PubMed2000

Assessment of arterial and cardiopulmonary baroreflex gains from simultaneous recordings of spontaneous cardiovascular and respiratory variability.

OBJECTIVES: In usual models of cardiovascular regulation, arterial pressure drives RR interval through a simple baroreflex, and the influence of respiration is dismissed. We examined the applicability of a trivariate autoregressive model to obtain separate values of the gain of the arterial and non-arterial, i.e. cardiopulmonary, components of the lumped baroreflex, employing spontaneous RR interval, systolic arterial pressure and respiration variability. DESIGN: We studied 30 normal subjects (age 37 +/- 1 years), both at rest and during standing, a condition known to enhance sympathetic activity while reducing venous return. Electrocardiogram was obtained by telemetry, arterial pressure by Finapres and respiration with a piezoelectric respiratory belt Data were acquired with a PC and processed with an ad hoc Windows program. METHODS: We employed an additive and a linear multivariate approach to approximate overall gain of the arterial pressure-heart beat period baroreflex (alphalumped) and of its arterial (alphaart) and non-arterial, i.e. cardiopulmonary (alphacp), components, from continuous beat-by-beat series of RR interval, systolic arterial pressure variability and respiration, without using any non-physiological intervention. RESULTS: The overall baroreflex gain at rest (alphalumped = 23.7 +/- 3.4 ms/mmHg) was subdivided into arterial (alphaart = 5.2 +/- 1.0 ms/mmHg) and cardiopulmonary (alphacp = 18.5 +/- 3.2ms/mmHg) components. During active orthostatism, alphaluumped was diminished to 10.0 +/- 2.2 ms/ mmHg. In addition, standing selectively reduced alphacp to 4.8 +/- 1.3 ms/mmHg, while alphaart was not significantly changed. CONCLUSIONS: A trivariate autoregressive model, that considers explicitly the influence of respiration, can subdivide overall, lumped, arterial pressure-heart period baroreflex gain, into two separate components, alphaart and alphacp. Only the latter is reduced by active orthostatism.

Adult↗

PubMed2000

Prediction of short cardiovascular variability signals based on conditional distribution.

A new approach measuring the predictability of a process is proposed. The predictor is defined as the median of the distribution conditioned by a sequence of L - 1 previous samples (i.e., a pattern). A function referred to as the corrected mean squared predictor error is defined to prevent the perfect adequacy to the data (i.e., the decrease to zero of the prediction error), thus avoiding to divide the whole set of data in learning and test sets. This function exhibits a minimum and this minimum is taken as a measure of predictability of the series. The use of the minimization procedure avoids to fix a priori the pattern length L. This approach permits one a reliable measure of predictability on short data sequences (around 300 samples). Moreover, this method, in connection with a surrogate data approach, is useful to detect nonlinear dynamics. The analysis indicates that, in simulated and real data, predictability and nonlinearity measures provide different information. The application of this approach to the analysis of cardiovascular variability series of the heart period (RR interval) and systolic arterial pressure (SAP) shows: 1) SAP series is more predictable than RR interval series; 2) predictability of the RR interval series is larger during tilt, during controlled respiration at 10 breaths/min (bpm) and after high-dose administration of atropine; 3) SAP series is dominated by linear correlation; 4) RR interval series exhibits nonlinear dynamics during controlled respiration at 10 bpm and after low-dose administration of atropine, while it is linear during sympathetic activation produced by tilt and after peripheral parasympathetic blockade caused by high-dose administration of atropine.

Adult↗

PubMed2000

Dominant active alleles of RIM101 (PRR2) bypass the pH restriction on filamentation of Candida albicans.

Morphological development of the fungal pathogen Candida albicans is profoundly affected by ambient pH. Acidic pH restricts growth to the yeast form, whereas neutral pH permits development of the filamentous form. Superimposed on the pH restriction is a temperature requirement of approximately 37 degrees C for filamentation. The role of pH in development was investigated by selecting revertants of phr2Delta mutants that had gained the ability to grow at acid pH. The extragenic suppressors in two independent revertants were identified as nonsense mutations in the pH response regulator RIM101 (PRR2) that resulted in a carboxy-terminal truncation of the open reading frame. These dominant active alleles conferred the ability to filament at acidic pH, to express PHR1, an alkaline-expressed gene, at acidic pH, and to repress the acid-expressed gene PHR2. It was also observed that both the wild-type and mutant alleles could act as multicopy suppressors of the temperature restriction on filamentation, allowing extensive filamentation at 29 degrees C. The ability of the activated alleles to promote filamentation was dependent upon the developmental regulator EFG1. The results suggest that RIM101 is responsible for the pH dependence of hyphal development.

Apoenzymes↗

PubMed2000

Assessing baroreflex gain from spontaneous variability in conscious dogs: role of causality and respiration.

A double exogenous autoregressive (XXAR) causal parametric model was used to estimate the baroreflex gain (alpha(XXAR)) from spontaneous R-R interval and systolic arterial pressure (SAP) variabilities in conscious dogs. This model takes into account 1) effects of current and past SAP variations on the R-R interval (i.e., baroreflex-mediated influences), 2) specific perturbations affecting R-R interval independently of baroreflex circuit (e.g., rhythmic neural inputs modulating R-R interval independently of SAP at frequencies slower than respiration), and 3) influences of respiration-related sources acting independently of baroreflex pathway (e.g., rhythmic neural inputs modulating R-R interval independently of SAP at respiratory rate, including the effect of stimulation of low-pressure receptors). Under control conditions, alpha(XXAR) = 14.7 +/- 7.2 ms/mmHg. It decreases after nitroglycerine infusion and coronary artery occlusion, even though the decrease is significant only after nitroglycerine, and it is completely abolished by total arterial baroreceptor denervation. Moreover, alpha(XXAR) is comparable to or significantly smaller than (depending on the experimental condition) the baroreflex gains derived from sequence, power spectrum [at low frequency (LF) and high frequency (HF)], and cross-spectrum (at LF and HF) analyses and from less complex causal parametric models, thus demonstrating that simpler estimates may be biased by the contemporaneous presence of regulatory mechanisms other than baroreflex mechanisms.

Animals↗

PubMed2000

Effects of spinal section and of positive-feedback excitatory reflex on sympathetic and heart rate variability.

The sympathetic outflow appears to be capable of displaying a rhythmicity synchronous with cardiovascular Mayer's waves even after spinal section. To test the hypothesis that spinal sympathetic low frequency (LF) oscillation can be enhanced during sympathetic excitation, we recorded cardiac sympathetic nerve activity (SNA), R-R interval, arterial pressure, and ventilation in 9 unanesthetized decerebrate-vagotomized cats before and after C1 spinal section. LF and high frequency (HF) components were detected in the variability of SNA, R-R interval, and systolic arterial pressure both before and after spinal section. In this latter condition, a significant coherence between LF(SNA) and LF(R-R) was present in 5 animals, whereas HF(SNA) and HF(R-R) were correlated in 4 animals. During an excitatory sympathetic spinal reflex elicited by aortic constriction, the efferent sympathetic firing was markedly enhanced (from 7+/-2 to 33+/-7 spikes/s); concomitantly, the powers of both LF(SNA) and HF(SNA) were also increased. Coherence between LF(SNA) and LF(R-R) became significant in all cases, whereas HF(SNA) and HF(R-R) became correlated in 6 animals. In 3 animals, the reflex sympathetic excitation was no longer elicitable after interrupting a vast contingent of sympathetic afferents by means of thoracic dorsal root section. We report for the first time that LF and HF oscillations are detectable in SNA, R-R interval, and systolic arterial pressure variabilities of decerebrate-vagotomized spinal cats and that an excitatory spinal reflex is capable of increasing the power of both SNA spectral components.

Animals↗

PubMed1999

Effect of calbindin D 28K on sodium transport by the luminal membrane of the rabbit nephron.

We previously reported that in the rabbit, the vitamin D-dependent calcium binding protein 28K (CaBP 28K) increases calcium (Ca2+) transport in the distal tubule by opening a high affinity Ca2+ channel in the luminal membrane. Since Na+ and Ca2+ transports are interdependent in this membrane, we questioned whether the calbindin has any influence on Na+ transport. Luminal membranes from rabbit proximal and distal tubules were purified and 22Na uptake by the membrane vesicles was measured using the rapid filtration technique. The vesicles were loaded with 280 mM mannitol and 20 mM Tris-Hepes pH 7.4, with either 3 microM CaBP or the carrier. Incubation medium contained 1 mM 22NaCl, 278 mM mannitol, and 20 mM Tris-Hepes pH 7.4. The presence of 3 microM CaBP 28K in the distal luminal membrane vesicles increased the 0.5 mM Ca2+ uptake from 0.91 +/- 0.21 to 1.84 +/- 0.33 pmol/microg/10 s (P < 0.01) and decreased 1 mM Na+ uptake from 0.62 +/- 0.15 to 0.27 +/- 0.08 pmol/microg/10 s (P < 0.05). A similar decrease of Na+ uptake was observed in proximal luminal membrane experiments. The effect on Na+ uptake by the distal membrane was dose-dependent with a IC50 of 4.5 microM. Addition of 2 mM Ca2+ to the incubation medium decreased 1 mM Na + uptake from 0.62 +/- 0.15 to 0.49 +/- 0.12 pmol/microg/10 s (P < 0.05), but did not influence the effect of CaBP 28K on Na+ uptake. Experiments performed in the presence and absence of ethyl isopropyl amiloride (EIPA) suggest that the effect of calbindin involves the Na+/H+ exchanger activity.

Animals↗

PubMed1999

An homologue of the human 100-kDa protein (p100) is differentially expressed by Histoplasma capsulatum during infection of murine macrophages.

Using differential display reverse transcription-PCR (DDRT-PCR) we have identified several sequences that are specifically expressed by Histoplasma capsulatum during infection of murine macrophages (MPhi). Here, we report the characterization of a clone, pHc12, identified as a differentially expressed gene 1 hour after infection of MPhi. Screening of a cDNA library of H. capsulatum allowed us to isolate a clone, pHc12-E, that contains the complete coding sequence. We show that after infection the level of transcription of this gene increases about 5 fold. Analysis of its sequence revealed the presence of an open reading frame of 890 aa (ORF890) that shares respectively 30 and 33% identity with human and Caenorhabditis elegans p100 kD and rat p105 kD co-activator proteins. Using the two-dimensional Hydrophobic Cluster Analysis (HCA) method, we showed that H. capsulatum ORF890 and p100 kD co-activator proteins are clearly related. The H. capsulatum protein consists of a four-fold repeated module (domains I to IV) like the p100 kD co-activator proteins, whose three-dimensional (3D) structure is related to staphylococcal thermonuclease, followed by a modified fifth "hybrid" domain which partially resembles the structure of the tudor domain found in multiple copies in the Drosophila melanogaster tudor protein. These data strongly suggest that ORF890 is homologous to human p100 kD and that this protein, named Hcp100, may play an essential role during infection by co-activating the expression of specific genes.

Amino Acid Sequence↗