Marine natural products as lead anti-HIV agents.

Current anti-HIV drugs have extreme side effects and resistance to these drugs develops rapidly. The marine environment holds an unprecedented number of unusual chemical structural classes with activity against HIV. We review the literature on anti-HIV activity of marine natural products and discuss the efficacy of different structural classes.

Enalapril improves vascular and cardiac function in streptozotocin-diabetic rats.

Recent evidence points to the renin-angiotensin system (RAS) as one of the systems involved in the etiology of micro- and macrovascular disease in diabetic patients. To help elucidate this possibility, the effect of daily treatment with enalapril (25 mg/kg/d) was evaluated in streptozotocin (STZ)-diabetic rats at 2 weeks following the induction of diabetes. Untreated diabetic rats and non-diabetic rats that were age-matched were used for comparison. Vascular studies included the determination of aortic ring responses to norepinephrine (NE), angiotensin II (Ang II) and acetylcholine (Ach). Systolic blood pressure (SBP), cardiac output (CO) indices, plasmatic and vascular angiotensin-converting enzyme (ACE) activity and thickness of the aortic wall were also assessed. Enalapril improved Ach-induced relaxation by increasing the maximal relaxation from 54.3 +/- 4.3% in untreated diabetic rats to 89.8 +/- 6.2% (n=9, p<0.05) and by decreasing the EC50 value from 32.6 +/- 9.9 nmol/l in untreated diabetic animals to 17.9 +/- 5.1 nmol/l (n=8, p<0.05). In addition, enalapril normalized the high responses to NE found in diabetic rats without inducing changes in the EC50 value. A significant reduction in SBP (from 158 +/- 4 mm Hg to 123 +/- 1 mm Hg, p<0.05), combined with an improved CO index (from 40 +/- 2 ml/min x 100 g BW to 50 +/- 1 ml/min x 100 g BW), was observed in the enalapril-treated diabetic group. A significant regression of the media thickness was also observed in the aorta of diabetic rats after treatment. ACE activity in the aorta of diabetic rats, that was doubled compared to controls (p<0.05), decreased after enalapril treatment. These results point to the vascular RAS as one of the key systems in the etiology of vascular alterations at early stages of diabetes. Therefore, ACE inhibitors, as well as other pharmacological approaches targeting the vascular RAS, should be considered in the treatment of diabetic patients from the very early stages of the condition.

Antimalarial (+)-trans-hexahydrodibenzopyran derivatives from Machaerium multiflorum.

Bioassay-guided fractionation of Machaerium multiflorum yielded the hitherto unreported (+)-trans-hexahydrodibenzopyrans machaeriol A (1) and machaeriol B (2), as well as the known guaiane sesquiterpene (-)-kessane. Structure elucidation was based on (1)H and (13)C NMR data, mainly 2D NMR (1)H-(1)H COSY, (1)H-(13)C HMQC, (1)H-(13)C HMBC, and (1)H-(1)H NOESY experiments. This is the first report of the hexahydrodibenzopyrans from a higher plant other than the genus Cannabis. The cannabimimetic activity was thus evaluated by radioligand binding assay for cannabinoid receptor CB1, which indicated, notably, that both 1 and 2 were inactive. In addition, the cross reactivity of 1 and 2 toward antibodies designed for urinary metabolites of cannabinoids was evaluated with the EMIT and On Line cannabinoids assays. Both compounds showed no response at 100 000 ng/mL in both assays. Machaeriol B (2) demonstrated in vitro antimalarial activity (IC(50) = 120 ng/mL) against Plasmodium falciparum W-2 clone.

A new naphthopyrone derivative from Cassia quinquangulata and structural revision of quinquangulin and its glycosides.

A novel naphthopyrone derivative, named quinquangulone (1), has been isolated from Cassia quinquangulata, along with the known compounds quinquangulin (2) and its two glycosides (3 and 4), rubrofusarin (5) and its two glycosides (6 and 7), nor-rubrofusarin (8) and its 6-O-glucoside (9), and three stilbenes (10-12). The structure of quinquangulone was established by spectral interpretation as 5,9-dihydroxy-8-methoxy-2,9-dimethyl-6-oxo-4H,6H,9H-naphtho-[2,3-b]pyran-4-one. Reinvestigation of the NMR spectra of quinquangulin led to revision of its structure as 5,6-dihydroxy-8-methoxy-2,9-dimethyl-4H-naphtho[2,3-b]pyran-4-one (2a). The structures of two quinguangulin glycosides, 3 and 4, were also revised accordingly. Compound 2a exhibited activity against Staphylococcus aureus and methicillin-resistant S. aureus (MIC, 3.125 and 6.25 microg/mL, respectively).

Indolopyridoquinazoline alkaloid from Leptothyrsa sprucei.

The indolopyridoquinazoline alkaloid, 3-hydoxyrutaecarpine, was isolated from Leptothyrsa sprucei, along with 8-methoxypsoralen, 5-methoxypsoralen, imperatorin, isoimperatorin, kaempferol 3-O-alpha-L-rhamnopyranoside, clematine and cnidioside B. The usefulness of the gradient 1H-15N HMBC NMR spectroscopy in the structure elucidation of 3-hydroxyrutaecarpine is noted.

New manzamine alkaloids with potent activity against infectious diseases.

The isolation of the new enantiomers of 8-hydroxymanzamine A (1), manzamine F (2), along with the unprecedented manzamine dimer, neo-kauluamine from an undescribed genus of Indo-Pacific sponge (family Petrosiidae, order Haplosclerida) is reported. The relative stereochemistry of neo-kauluamine was established through detailed analysis of NOE-correlations combined with molecular modeling. The significance of the manzamines as in vivo antimalarial agents with superior activity to the clinically used drugs artemisinin and chloroquine is discussed along with the activity in vitro against the AIDS-opportunistic infectious diseases tuberculosis and toxoplasmosis. Reexamination of the sponges identified as Prianos, and Pachypellina, in earlier publications has confirmed that these are members of the same genus as the sponge described here, but differ at the species level.

Investigation of Uña De Gato I. 7-Deoxyloganic acid and 15N NMR spectroscopic studies on pentacyclic oxindole alkaloids from Uncaria tomentosa.

The C-8-(S) isomer of deoxyloganic acid (7-deoxyloganic acid), together with beta-sitosteryl glucoside, five known stereoisomeric pentacyclic oxindole alkaloids and the tetracyclic oxindole isorhyncophylline, were isolated from the inner bark of Uncaria tomentosa. Structures of the isolated compounds were based on 1H and 13C NMR data, mainly 2D NMR experiments, including 1H-13C HMBC and 1H-1H NOESY correlation. Furthermore, the hitherto unreported 15N chemical shifts of the isomeric oxindole alkaloids, using 1H-15N HMBC experiments, were utilized to facilitate their characterization. Uncarine D showed weak cytotoxic activity against SK-MEL, KB, BT-549 and SK-OV-3 cell lines with IC(50) values between 30 and 40 microg/ml, while uncarine C exhibited weak cytotoxicity only against ovarian carcinoma (IC(50) at 37 microg/ml).

Antimalarial, cytotoxic, and antifungal alkaloids from Duguetia hadrantha.

Bioassay-guided isolation of Duguetia hadrantha yielded two new 4,5-dioxo-1-azaaporphinoids, hadranthine A (1) and hadranthine B (2), together with the known alkaloids imbiline-1 (3), sampangine (4), and 3-methoxysampangine (5), whose structures were determined primarily from 2D-NMR 1H-13C HMBC, and 1H-15N HMBC experiments. This is the first report of the co-occurrence of the copyrine alkaloids 4 and 5, as well as the first report of either copyrine or imbiline type alkaloids from a Duguetia species. Compounds 1, 4, and 5 demonstrated in vitro antimalarial activity against Plasmodium falciparum (W-2 clone), while 2 was inactive. Instead, 2 showed in vitro cytotoxicity to selected human cancer cell lines (IC50 = 3-6 microg/mL against SK-MEL, KB, BT-549, and SK-OV-3), and 4 was also cytotoxic to human malignant melanoma (IC50 = 0.37 microg/mL). Sampangine (4) also inhibited cell aggregation with a MIC value of <0.15 microg/mL, while 3-methoxysampangine (5) was only weakly active.

Transformation of jervine by Cunninghamella elegans ATCC 9245.

Preparative-scale fermentation of the known C-nor-D-homosteroidal jerveratrum alkaloid jervine with Cunninghamella elegans (ATCC 9245) has resulted in the isolation of (-)-jervinone as the major metabolite. In addition, C. elegans ATCC 9245 was able to epimerize C-3 of jervine, producing 3-epi-jervine. This epimerization reaction was similar to that reported for tomatidine, the known spirosolane-type Solanum alkaloid. The structure elucidation of both metabolites was based primarily on 1D- and 2D-NMR analyses.

Development of posture and locomotion in free-ranging primates.

Postural and locomotor development is described for free-ranging groups of three monkey species: Macaca mulatta (rhesus macaque), M. radiata (bonnet macaque) and Presbytis entellus (hanuman langur). Behaviors are discussed in terms of infant-1, infant-2, juvenile, adult and elderly age groups. Analysis is based on high-speed motion pictures and videos. Infant-1 monkeys continually cling to their mother's ventral surface with strong hand and foot grasps. Independent motor coordination develops during the infant-2 period. The greatest postural and locomotor activity, both qualitatively and quantitatively, is achieved during the juvenile period. The adult and elderly periods are characterized by a progressive reduction in the variety and, eventually, quality of these motor abilities. Macaques and langurs differ, however, in the course of their behavioral development. These findings have implications for concepts of underlying neural mechanisms, indicate possible species differences in the development of these mechanisms, and demonstrate that the spectrum of volitional behaviors practiced by different age groups of free-ranging animals is broader than that observed in the laboratory setting.

Activity of neuromuscular compartments in lateral gastrocnemius evoked by postural corrections during stance.

1. The electromyographic (EMG) activity of the four neuromuscular compartments in lateral gastrocnemius (LG) of cats was investigated to determine whether these intramuscular subdivisions could be activated differentially during automatic postural corrections. EMG electrodes were surgically implanted into each of the four compartments of left LG-LG1, LG2, LG3, and LGm--in two cats. Electrodes were also implanted into soleus and gluteus medius for comparative purposes. 2. Quiet quadrupedal stance was disturbed first by linearly translating the cats on a movable platform in each of 16 different horizontal directions. Mechanical events during corrections were characterized in terms of the three-dimensional forces exerted by each paw on the platform. EMG and force traces were quantified (area under the curve) and normalized, and tuning curves were constructed that relate muscle response and force change to direction of platform movement. 3. In a second series of trials, translations were presented along one direction only over a series of six velocities ranging from 5 to 16 cm/s. The third series of perturbations, termed the pop-up, consisted of a rapid upward displacement of the support under the left hindlimb only over a series of six amplitudes ranging from 1 to 10 mm. Evoked EMG activity and average change in force were normalized and regressions were computed onto velocity and amplitude, respectively. The slopes of the regressions were compared. 4. EMG tuning curves associated with the multidirectional horizontal translations revealed no differential activity across LG compartments. Similarly, there was no statistical difference among the slopes of the regressions within LG. In contrast, soleus exhibited significantly different slopes from LG for the regressions. Thus it is concluded that LG compartments are not differentially activated during automatic postural responses to perturbations of the support surface.

Locomotor behavior of rhesus macaques (Macaca mulatta) on Cayo Santiago.

The locomotor behaviors of Cayo Santiago rhesus monkeys (Macaca mulatta) were studied in order to determine qualitatively some of the similarities and differences in movement patterns between free-ranging and captive primates. Super-8mm cinefilms of locomotor sequences by all age groups were analyzed frame-by-frame. Cayo Santiago macaques use locomotor behaviors on the ground, large-diametered branches, and vines that correspond to captive monkey locomotion on comparable supports (floors, suspended poles, ropes). Cayo Santiago macaques differ from captive monkeys, however, in that most infants begin to walk using the diagonal-sequence pattern, swimming is common, and infants and juveniles practice a variety of locomotor behaviors among the small-diametered tree branches. Therefore, the Cayo Santiago facility provides 1) insights into the full repertoire of infant and juvenile locomotor behaviors that are essential for studies of motor development and its neural control, and 2) models for designing small-branch supports for captive colonies.

Stance dependence of automatic postural adjustments in humans.

This study investigated the effect of initial stance configuration on automatic postural responses in humans. Subjects were tested in both bipedal and quadrupedal stance postures. The postural responses to horizontal translations of the supporting surface were measured in terms of the forces at the ground, movement of the body segments, and electromyographic (EMG) activity. Postural responses to the same perturbations changed with initial stance posture; these responses were biomechanically appropriate for restoring centre of mass. A change in stance configuration prior to platform movement led to a change in both the spatial and temporal organization of evoked muscle activation. Specifically, for the same direction of platform movement, during bipedal stance muscles on one side of the lower limb were activated in a distal to proximal sequence; during quadrupedal stance, muscles on the opposite side of the lower limb were activated and in a proximal to distal sequence. The most significant finding was an asymmetry in the use of the upper limbs and the lower limbs during postural corrections in quadrupedal stance. Whereas antagonists of the upper limb were either co-activated or co-inhibited, depending on the direction of translation, lower limb antagonists were reciprocally activated and inhibited. Human subjects in a quadrupedal stance posture used the lower limbs as levers, protracting or retracting the hips in order to propel the trunk back to its original position with respect to the hands and feet. Postural responses of the subjects during quadrupedal stance were remarkably similar to those of cats subjected to similar perturbations of the supporting surface. Furthermore, the same predominance of lower limb correction is characteristic of both species, suggesting that the standing cat is a good model for studying postural control in humans.

Automatic postural responses in the cat: responses of proximal and distal hindlimb muscles to drop of support from a single hind- or forelimb.

Cats respond to drop of the support from beneath a single limb with the "diagonal stance response" (Coulmance et al. 1979). They load the limbs on the diagonal opposite to the one containing the dropped limb and unload the third supporting limb in the diagonal containing the dropped limb. Characteristic biomechanical delays in limb motion and in vertical force changes imposed upon the limbs are observed. These delays range from 30 to 45 ms, depending upon the location of the dropped limb. This study describes the kinematics of the "diagonal stance response" and the activation of selected agonist-antagonist muscle pairs acting on the joints of the hindlimb during the response. Proximal and distal hindlimb muscles respond to perturbations in groups that are appropriate to the vertical forces imposed upon the limb. When the hindlimb containing the recording electrodes is loaded by drop of the contralateral hindlimb or the ipsilateral forelimb medium latency (25-45 ms) EMG responses occur in the extensors. This response serves to stiffen the limb against the increased vertical force of loading. A similar response is observed when the hindlimb is reloaded after being dropped. In this case, however, short latency responses precede the medium latency responses in muscles that are passively stretched by the limb drop. When drop of the diagonal forelimb unloads the hindlimb containing the electrodes, medium latency responses are observed in the distal hindlimb flexors, which indicates that the unloading is evoked in part by active lifting of the limb. In most cases, the medium latency responses precede or are coincident with the changes in force imposed on the limb, suggesting that the observed responses are centrally programmed.

Automatic postural responses in the cat: responses of distal hindlimb muscles to paired vertical perturbations of stance.

The active components of the quadrupedal diagonal stance response to rapid removal of the support from beneath a single limb were studied in cats to further define the mechanisms that trigger and generate the response. We recorded EMG activity from lateral gastrocnemius and tibialis anterior muscles in awake, behaving cats while they stood on an hydraulic posture platform. By dropping the support from beneath a single limb, we evoked the diagonal stance response, with its characteristic changes in vertical force and EMG patterns. As the animal responded to this drop, a second perturbation of posture was then presented at intervals of 10 to 100 ms following the first. The second perturbation, which consisted of dropping the support from beneath the two limbs that were loaded as a result of the initial limb drop, made the first response biomechanically inappropriate. The EMG responses observed in both muscles during paired perturbations were triggered by the somatosensory events related to the perturbations. Muscle responses that were appropriate for the first perturbation always occurred with amplitudes and latencies similar to control trials. This was true even when the second perturbation occurred 10-20 ms after the first, that is, when this perturbation either preceded or was coincident with the response to the initial limb drop. The EMG responses that were normally associated with the second perturbation were delayed and/or reduced in amplitude when the time interval between perturbations was short. As the inter-perturbation interval was lengthened beyond 60-100 ms, however, EMG responses to the second perturbation were unaffected by the occurrence of the first perturbation. When the hindlimb containing the recording electrodes was dropped as part of the second perturbation, a myotatic latency response was observed in tibialis anterior. The amplitude of this response to the second perturbation was greater than controls when this displacement was presented during the period between initiation of the first perturbation and execution of the response to it. When the second displacement was presented after execution of the first response began, the amplitude of the myotatic response was reduced below control levels. While the results do not preclude the possibility that these "automatic" postural responses are segmental or suprasegmental reflexes, they support the hypothesis that the active component of the response to drop of the support beneath a single limb is centrally programmed and that the appropriate response can be triggered very rapidly by the somatosensory information signalling the perturbation.

Postural responses in the cat to unexpected rotations of the supporting surface: evidence for a centrally generated synergic organization.

Postural reactions to disruptions of stance are rapid and automatic in both quadrupeds and bipeds. Current evidence suggests that these postural responses are generated by the central nervous system as patterns involving muscle synergies. This study attempted to test this hypothesis of a centrally generated postural mechanism by determining whether the same postural response could be evoked in the freely-standing cat under two different biomechanical conditions. The present work is an extension of previous experiments in which the stance of cats was perturbed by a horizontal translation of the supporting surface in the anterior and posterior directions (Rushmer et al. 1983). We now tested whether simple rotation of the metacarpo- and metatarsophalangeal (M-P) joints that mimics the digit rotation occurring during platform translation, was sufficient to evoke the translation postural response. The rotational perturbations were biomechanically different from translations in that the rotation did not cause displacement of the centre of mass of the animal, nor did it result in any significant movement about any but the M-P joints. Even so, rotational perturbations did evoke the appropriate translational muscle synergies in all four animals. Both plantar flexion rotation and headward translation activated the posterior hindlimb synergy (which included gluteus medius, semitendinosus and lateral gastrocnemius). Similarly, dorsiflexion rotation and tailward translation both activated the same anterior hindlimb synergy (iliopsoas, vastus lateralis and tibialis anterior) together with the forelimb synergy The postural responses elicited by rotational perturbations were biomechanically inappropriate, and caused the animal to displace its own centre of mass away from the stable, control position.(ABSTRACT TRUNCATED AT 250 WORDS)

Neural control of quadrupedal and bipedal stance: implications for the evolution of erect posture.

The transition among hominids from quadrupedalism to bipedalism resulted in modifications in their musculoskeletal morphology. It is unclear, however, whether changes in the circuitry of the CNS were also necessary in order to accommodate the unique balance requirements of two-limb support. This study addresses the issue of modifications in control strategies by investigating the rapid, automatic postural responses of feline and human subjects to sudden disturbances of balance in the anteroposterior (AP) direction while they stand quadrupedally and bipedally on movable platforms. Postural responses are characterized in terms of segmental adjustments, generated AP shear forces, and electromyographic activity. Feline and human subjects correct posture similarly when standing quadrupedally. Furthermore, both species correct stance primarily with their hindlimbs and use their forelimbs as supportive struts. In contrast, both species use completely different correctional strategies when standing bipedally. Morphological restrictions, however, prevent cats from adopting the pillar-like plantigrade posture of human beings. Thus, the correctional strategies of bipedal cats are distinct from those of bipedal human subjects. It is concluded that 1) automatic postural response patterns of quadrupedal Felis and bipedal Homo reflect the different biomechanical characteristics of the initial postures rather than species differences in CNS circuitry controlling stance; 2) hindlimb-dominated posture control is probably a common and relatively ancient pattern; and 3) reorganization of hominid CNS circuitry was probably unnecessary because hindlimb control was already a feature of the system.

Automatic postural responses in the cat: responses to headward and tailward translation.

EMG responses, vertical and A-P shear forces and kinematics of "automatic postural responses" to unexpected translational perturbations in the headward and tailward directions were studied in cats. Muscles acting on the major joints of the forelimbs and hindlimbs were studied. Movement of the animals in response to perturbation were highly stereotyped and consisted of two phases: (1) motion of the feet during platform movement while the trunk remained relatively stationary followed by (2) active correction of posture by movement of the trunk in the direction of perturbation. Vertical force changes occurred after the perturbation was well underway (latency 65 ms) and were related to the displacement of the center of mass and active correction of trunk position. Shear forces showed both passive (inertial) and active components and suggested that the majority of the torque necessary for postural correction was generated by the hindlimb. EMG responses in forelimb and shoulder muscles were most correlated with increase in vertical force, showing a generalized co-contraction in tailward translation (when these limbs were loaded) and little activity when the forelimbs were unloaded. EMG responses in hindlimb showed reciprocal activation of agonists and antagonists during perturbation with strong synergies of thigh and foot flexors in tailward translation and thigh and foot extensors in headward translation. The forelimb EMG patterns were most consistent with the conclusion that the forelimb is used primarily for vertical support during perturbation. It was concluded that hindlimb EMG responses were appropriate for both vertical support and performance of the postural correction. The hindlimb muscle synergies observed during translation are the "mirror image" of those observed in humans by other workers.