Topical treatment for human papillomavirus-associated genital warts in humans with the novel tellurium immunomodulator AS101: assessment of its safety and efficacy.

BACKGROUND: Various methods are currently used for the treatment of anogenital warts. However, a complete cure is unlikely, and the rate of recurrence is high. OBJECTIVES: The purpose of this open-label, multicentre trial was to evaluate the safety and clinical efficacy of a new treatment using the immunomodulator ammonium trichloro (dioxoethylene-O,O') tellurate (AS101; Biomas Ltd, Kefar Saba, Israel) 15% w/w cream to clear vulval/perianal condylomata acuminata. METHODS: Study participants comprised 48 women and 26 men, age range 18-62 years. Of the 48 woman, 44 were diagnosed with vulval condylomata and four with perianal condylomata. All 26 men were diagnosed with perianal condylomata. All the patients in the study received AS101 15% w/w cream twice a day. Maximal treatment duration was 16 weeks. To evaluate the safety and clinical efficacy, patients were examined and lesional areas photographed on a biweekly basis. RESULTS: By the end of the treatment, 56 of 74 (76%) patients were considered completely cleared. Complete cure was achieved in 35 of 44 (80%) patients with vulval condylomata and in 21 of 30 (70%) patients with perianal condylomata. No scarring of treated areas was observed. Complete cure was achieved within a time range of 10-109 days. The most frequent side-effects observed were mild-to-moderate itching, soreness, burning and erythema. In post-treatment follow up of up to 6 months, disease recurrence was observed in two patients (4%), at 105 and 144 days following completion of treatment. CONCLUSIONS: AS101 15% w/w cream is an effective and safe, self-administered therapy used for the treatment of external vulval and perianal warts. The cream is applied topically twice daily for up to 16 weeks. A very low recurrence rate was reported.

A novel phospholipid derivative of indomethacin, DP-155 [mixture of 1-steroyl and 1-palmitoyl-2-{6-[1-(p-chlorobenzoyl)-5-methoxy-2-methyl-3-indolyl acetamido]hexanoyl}-sn-glycero-3-phosophatidyl [corrected] choline], shows superior safety and similar efficacy in reducing brain amyloid beta in an Alzheimer's disease model.

Indomethacin has been suggested for the treatment of Alzheimer's disease (AD), but its use is limited by gastrointestinal and renal toxicity. To overcome this limitation, D-Pharm Ltd. (Rehovot, Israel) developed DP-155 (mixture of 1-steroyl and 1-palmitoyl-2-{6-[1-(p-chlorobenzoyl)-5-methoxy-2-methyl-3-indolyl acetamido] hexanoyl}-Sn-glycero-3-phosophatidyl [corrected] choline), a lecithin derivative of indomethacin. Safety was tested by daily oral administration of DP-155 or indomethacin to rats in a dose range of 0.007 to 0.28 mmol/kg. The prevalence of gastrointestinal ulceration was significantly lower (10-fold) for DP-155 than for indomethacin, and the ulcerations were delayed. Signs of renal toxicity, namely reduced urine output and increased urine N-acetyl glycosaminidase to creatinine ratio, were 5-fold lower for DP-155. Indomethacin, but not an equimolar dose of DP-155, reduced urine bicyclo-prostaglandin E(2). An equimolar oral dose of DP-155 or indomethacin, administered every 4 h for 3 days, was equally efficacious in reducing the levels of Abeta42 in the brains of Tg2576 mice. Indomethacin was the principal metabolite of DP-155 in the serum. After DP-155 oral administration, indomethacin's half-life in the serum and the brain was 22 and 93 h, respectively, compared with 10 and 24 h following indomethacin oral administration. The brain to serum ratio was 3.5 times higher for DP-155 than indomethacin. This finding explains the efficacy of DP-155 in reducing Abeta42 brain levels, despite the low systemic blood concentrations of indomethacin derived from DP-155. In conclusion, compared with indomethacin, DP-155 has significantly lower toxicity in the gut and kidney while maintaining similar efficacy to indomethacin in lowering Abeta42 in the brains of Tg2576 mice. This superior safety profile highlights DP-155's potential as an improved indomethacin-based therapy for AD.

Intraarterial delivery of genetic vectors for the treatment of malignant brain tumors.

BACKGROUND: The transfer of therapeutic genes into malignant brain tumors has been the subject of intense preclinical and clinical research in recent years. Most approaches have used direct intratumoral placement of a variety of vectors and genes, such as retroviruses or adenoviruses carrying drug-susceptibility genes, modified replication-competent herpes virus, and several vectors carrying tumor suppressor genes such as the p53 gene. However, clinical results have so far been disappointing, mainly due to the limited ability to effectively distribute the genetic material into the target cell population. Accordingly, alternative delivery approaches into the central nervous system, e.g., intravascular, are under investigation. Genetic vectors administered intravascularly are unlikely to penetrate the blood-brain barrier and transfer a gene into brain or tumor parenchyma. However, intravascular delivery of vectors may target endothelial cells lining the blood vessels of the brain. Since endothelial cells participate in a variety of physiological and pathological processes in the brain, their modulation by gene transfer may be used for a variety of therapeutic purposes. Angiogenically stimulated endothelial cells within tumors replicate rapidly and hence may become targets for retroviral-mediated gene transfer. OBJECTIVE: To assess the anti-tumor effect of transferring a drug-susceptibility gene into endothelial cells of the tumor vasculature. METHODS: As a model for this approach we delivered concentrated retroviral vectors carrying a drug-susceptibility gene via the internal carotid artery of rats with malignant brain tumors. The safety and efficacy of this approach, without and with subsequent treatment with a pro-drug (ganciclovir), was evaluated. RESULTS: No acute or long-term toxicity was observed after intraarterial infusion of the vector. Treatment with ganciclovir resulted in variable hemorrhagic necrosis of tumors, indicating preferential transduction of the angiogenically stimulated tumor vasculature. This was accompanied by severe toxicity caused by subarachnoid hemorrhage and intracerebral hemorrhage in vascular territories shared by the tumor and adjacent brain. CONCLUSION: The data indicate that endothelial cells can be targeted by intraarterial delivery of retroviral vectors and can be used for devising new gene therapy strategies for the treatment of brain tumors.

Progesterone rapidly decreases brain edema: treatment delayed up to 24 hours is still effective.

Cerebral edema is a serious side effect of traumatic brain injury. We have previously established that progesterone injections, initiated within 1 h after cortical contusion injury, reduced edema when assessed 3 days later. To determine how rapidly progesterone can reduce edema, male and female rats were given the hormone 1 h after damage to the medial frontal cortex, and edema levels were assessed between 2 h and 7 days postinjury. Progesterone decreased edema with 6 h of the injury and continued to be effective for the duration of treatment. In addition, we assessed whether progesterone injections are effective when delays are imposed between injury and initiation of treatment. Male and female rats received progesterone after postinjury delays 6, 24, or 48 h. Progesterone was effective in reducing edema when treatment was delayed until 24 h after injury.

Blood-brain barrier breakdown and edema formation following frontal cortical contusion: does hormonal status play a role?

The present experiment was designed to evaluate and correlate the time course of blood-brain barrier (BBB) integrity and cerebral edema in adult male rats given medial frontal cortex contusions. The effect of sex hormones on BBB integrity in the same injury model was also examined, because previous work has shown that progesterone can reduce cerebral edema (Roof et al., 1993). BBB breakdown was assessed by Evans blue extravasation and albumin immunostaining while edema formation was measured by the wet weight dry weight technique. These processes were examined beginning 2 h and continuing up to 10 days after injury. Our findings show that medial frontal contusion in rats produces changes in cerebral water content and opening of the BBB that endures at least 7 days postinjury. Although pseudopregnancy has been shown to reduce cerebral edema at day 1 postinjury, we did not find any evidence that this hormonal state is associated with BBB repair.

Progesterone facilitates cognitive recovery and reduces secondary neuronal loss caused by cortical contusion injury in male rats.

The ability of progesterone to reduce the cerebral edema associated with traumatic brain damage first became apparent when we observed that males had significantly more edema than females after cortical contusion. In addition, edema was almost absent in pseudopregnant female rats, a condition in which progesterone levels are high relative to estrogen. Progesterone injections given after injury also reduced edema and were equally effective in both males and females. The present experiment was done to determine if the progesterone-induced reduction in edema could also prevent secondary neuronal degeneration and reduce the behavioral impairments that accompany contusion of the medial frontal cortex. Progesterone-treated rats were less impaired on a Morris water maze spatial navigation task than rats treated with the oil vehicle. Progesterone-treated rats also showed less neuronal degeneration 21 days after injury in the medial dorsal thalamic nucleus, a structure that has reciprocal connections with the contused area.

A new method for expressing axonal size: rat optic nerve analysis.

Analysis of the shape of the cross sections of adult rat optic nerve axons reveals that the majority of axons do not have a true circular shape. Therefore, determination of axonal size has to utilize methods of approximation. The method presented here utilizes three calculated parameters for expression of axonal size: (i) axonal diameter, as calculated from its area, or (ii) axonal diameter, as calculated from its perimeter, both assuming axonal shape to be a perfect circle and (iii) axonal shape factor, which represents the divergence of the axon from a perfect circular shape. The use of the calculated axonal diameter, with a correction for its shape factor, provides a normalized way of expressing axonal size.

Gender influences outcome of brain injury: progesterone plays a protective role.

The contributions of gender and gonadal hormones in the cascade of events following brain injury are largely unexplored. We measured cerebral edema following cerebral contusion in rats under three hormonal conditions to address this issue. Normally cycling females exhibited significantly less edema than males, and pseudopregnant females were virtually spared from post-injury edema. Subsequent studies of ovariectomized females, with or without hormone treatment, indicated that the reduction of cerebral edema was associated primarily with the presence of circulating progesterone. We conclude that progesterone has a protective effect on the brain following traumatic injury.

Gangliosides attenuate axonal loss after optic nerve injury.

Gangliosides have been shown to be capable of protecting nerve tissue from mechanical and biochemical insults and promoting their repair. The present study provides morphologic evidence that monosialogangliosides attenuate the degenerative process at the distal stump of the rat optic nerve after crush injury. Injured rat optic nerves were treated for 7 days after injury with daily intraperitoneal injections of monosialogangliosides (30 mg/kg/day), and compared with untreated injured controls with respect to the number of viable axons 2 and 4 weeks after injury, as indicated by transmission electron microscopy. After 2 weeks, the mean number of viable axons in the treated optic nerves (n = 5) was slightly higher than in the controls (n = 5). Four weeks after injury, although the absolute number in both the experimental and the control groups had dropped, it was about seven-fold higher in the treated animals (1696 +/- 1149, n = 7) than in the untreated animals (216 +/- 65, n = 6); this difference was statistically significant. These findings, which offer some insight as to how monosialogangliosides affect injured nerves, may have important implications for treatment in cases of optic nerve injury.

Dose and temporal parameters in delaying injured optic nerve degeneration by low-energy laser irradiation.

Low-energy laser irradiation has been reported to postpone the degenerative processes in crushed optic nerves of rats, which are part of the nonregenerable mammalian central nervous system. In the present study, we evaluated the optimal irradiation parameters for this purpose. Optic nerves of 141 rats were subjected to crush injury and then irradiated through the eye, starting at different points of time before or after the injury, for different durations and periods, using various intensities of either helium-neon laser or noncoherent infrared light (904 nm). The effect was evaluated by measurements of the compound action potentials of the nerve segments between the site of injury and the optic chiasm. The compound action potential amplitude of the crushed nonirradiated nerves, as measured 2 weeks after the injury, was found to be 0.51 +/- 0.30 mV, in contrast to 3.10 +/- 1.03 mV measured in 232 normal nerves. Irradiation with a 10.5 mW helium-neon laser for 2 and 3 min once a day for 14 consecutive days resulted in maximal preservation of action potentials (1.78 +/- 0.72 and 1.95 +/- 0.71 mV, respectively). Irradiations beginning immediately prior to the injury were as effective as irradiations beginning soon after it. Irradiations for longer than 3 min or twice a day aggravated the damage. Noncoherent infrared light was ineffective or adversely affected the injured nerves. Our experiments suggest that optimal delay of posttraumatic optic nerve degeneration in rats is attainable with 10.5 mW helium-neon laser irradiations for 2 or 3 min once a day for 14 consecutive days.

GM1 reduces injury-induced metabolic deficits and degeneration in the rat optic nerve.

This study demonstrates the earliest reported effects of GM1 treatment on crush-injured axons of the mammalian optic nerve. GM1, administered intraperitoneally immediately after injury, was found to reduce the injury-induced metabolic deficit in nerve activity within 2 hr of injury, as measured by changes in the nicotine-amine adenine dinucleotide redox state. After 4 wk, transmission electron microscopy 1 mm distal to the site of injury revealed a sevenfold increase in axonal survival in GM1-treated compared to untreated injured nerves. These results emphasize the beneficial effect of GM1 on injured optic nerves as well as the correlation between immediate and long-term consequences of the injury. Thus, these results have implications for treating damaged optic nerves.

Disappearance of astrocytes and invasion of macrophages following crush injury of adult rodent optic nerves: implications for regeneration.

Injury to the mammalian central nervous system results in loss of function because of its inability to regenerate. It has been postulated that some axons in the mammalian central nervous system have the ability to regenerate but fail to do so because of the inhospitable nature of surrounding glial cells. For example, mature oligodendrocytes were shown to inhibit axonal growth, and astrocytes were shown to form scar tissue that is nonsupportive for growth. In the present study we report an additional phenomenon which might explain the failure of axons to elongate across the site of the injury, namely, the absence of astrocytes from the crush site between the glial scar and the distal stump. Astrocytes began to disappear from the injury site as early as 2 days after the injury. After 1 week the site was necrotic and contained very few glial cells and numerous macrophages. Disappearance of glial cells was demonstrated in both rabbit and rat optic nerves by light microscopy, using antibodies directed against glial fibrillary acidic protein, and by transmission electron microscopy. Results are discussed with reference to possible implications of the long-lasting absence of astrocytes from the injury site, especially in view of the differences between the present findings in rodents and our recent observations in fish.

Progesterone treatment attenuates brain edema following contusion injury in male and female rats.

To assess the effectiveness of progesterone as a treatment for edema following traumatic brain injury, male and female rats were injected with progesterone or the oil vehicle over a 3-day period following a cortical contusion injury. Oil-treated rats showed significant localized edema as measured by the wet weight/dry weight tissue punch technique. Progesterone-treated rats, both male and female, showed marked attenuation, or in some cases, absence of this post-injury edema. We conclude that progesterone shows promise as a treatment for edema following traumatic brain injury.

GM1 ganglioside treatment reduces visual deficits after graded crush of the rat optic nerve.

Despite numerous reports of beneficial effects of GM1 ganglioside treatment following brain lesions in animals, the underlying neurobiological mechanism of ganglioside-induced functional restoration is still unclear. In order to obtain a better insight into this question, we have made use of a newly developed animal model of brain injury that would potentially permit us to determine the causal relationship(s) among behavioral and neuroanatomical/neurochemical parameters of restoration of function. Following graded crush of the adult rat optic nerve, we have treated the rats with intraperitoneally injected gangliosides and studied the functional outcome with electrophysiological and behavioral parameters. The electrophysiological recording of the compound action potential (CAP) from excised rat optic nerve revealed a significant loss of CAP throughout the first 2 weeks after the injury. However, when rats were treated daily for 7 days with GM1-gangliosides, the CAP measured 10 days after the crush was significantly larger compared to operated controls without treatment. Thus, GM1 appeared to be capable of delaying or partially preventing retinal ganglion cells or their axons from secondary degeneration. Loss of visual function was also evident on the behavioral level of analysis: when rats with unilateral optic nerve crush were evaluated in a visual orienting paradigm, the rats revealed deficits in their ability to orient towards small, moving visual stimuli. However, within about 2 weeks, the animals recovered spontaneously to near normal performance. Daily treatment with GM1-gangliosides was found to significantly improve outcome, largely due to a reduction of the immediate post-lesion deficit. In a second behavioral experiment we also created graded crush in rats bilaterally and evaluated the animals visual capacities in a two-choice brightness discrimination task. In this task, an initial loss of function was followed by recovery within about 2 weeks, but GM1 treatment was without beneficial effects in this paradigm. It is concluded that GM1 improves outcome after graded crush of the adult rat optic nerve, although it appears that improved function needs to be documented with sufficiently sensitive behavioral assays.

Oligodendrocyte cytotoxic factor associated with fish optic nerve regeneration: implications for mammalian CNS regeneration.

The limited capacity for regenerative axonal growth by adult mammalian central neurons has been attributed, at least in part, to the presence of mature oligodendrocytes, which are non-permissive for axonal growth. These cells do not interfere with growth during development, as developmental growth is largely completed before the maturation of the oligodendrocytes. Unlike mammals, fish central nervous system is endowed with a high regenerative capability. When soluble substances derived from regenerating fish optic nerves are applied to injured adult rabbit optic nerves, regenerative axonal growth is permitted. Therefore, in the present study, we tested whether the fish optic nerve, after injury, is endowed with a mechanism by which it avoids the possible inhibitory effect of the process-bearing mature oligodendrocytes. Specifically, we looked for the possible presence of soluble substances that can regulate the number of process-bearing mature oligodendrocytes. We found that soluble substances derived from regenerating fish optic nerve, when added to cultures of oligodendrocytes derived from newborn or injured adult rat optic nerves, caused a decrease in the number of process-bearing mature oligodendrocytes. Soluble substances derived from normal noninjured fish optic nerves, had a significantly lower effect. The observed decrease in the number of mature oligodendrocytes could not be mimicked by the addition of platelet-derived growth factor (PDGF), a known mitogen of oligodendrocyte progenitors which transiently inhibits their maturation. This study suggests a role to oligodendrocyte inhibitory/cytotoxic factor(s) in regeneration.

Glial response to axonal injury: in vitro manifestation and implication for regeneration.

Crushed fish optic axons readily regenerate, while similarly injured rat optic axons do not; the reasons for the differences in regeneration ability may lie in differences in the environment of the axons. We have cultured glial cells from previously crushed optic nerves of fish and rat to determine whether a relationship exists between the ability to regenerate and the nature of the responses of the associated nonneuronal cells to injury. The glial cells were examined using indirect immunofluorescence with antibodies to known glial markers. In the rat cultures, mature GalC oligodendrocytes, which are known to be nonpermissive for axonal growth, were abundant. In contrast, in the fish cultures mature oligodendrocytes were rare, but A2B5 positive cells were abundant. The high number of A2B5 positive cells in the fish may suggest a high number of immature cells. This interpretation, however, should wait until evidence for glial cell lineage of the fish is available. Additional indication is provided also in the present study that the number of mature oligodendrocytes in the fish is regulated by elements external to the nerve. This study thus demonstrates an important difference between rat and fish optic nerves in the response of glial cells to the optic nerve injury.

Graded crush of the rat optic nerve as a brain injury model: combining electrophysiological and behavioral outcome.

In the present study, we have developed an animal model for central nervous system (CNS) damage using the graded crush injury of the adult rat optic nerve. This injury model has the following characteristics: (a) the injuries are reproducible; (b) the injuries can be created with controlled severity; and (c) variations in the severity of the injuries correlate with definable electrophysiological and behavioral outcomes. Self-closing forceps, modified by attaching a screw to the handle, were used as a lesion-causing device. Rat optic nerves were crushed in vitro. The severity of the injury, calibrated by a measured applied force, was determined, electrophysiologically, in vitro. Compound action potential was assessed and recorded by suction electrodes connected to both sides of the nerve. In vivo studies correlated the calibrated crush injuries with behavioral outcome. After severe crush (applied force: 205 g or higher), the rats were unable to orient towards visual stimuli presented in the visual field of the damaged nerve and only minor transient recovery was observed. After moderate crush (applied force: 120 g), however, there was a significant recovery of orienting performance up to the 6th postoperative day, followed by a gradual, secondary loss of function. In the mildly injured group (applied force lower than 30 g), functional improvement continued for up to 10 days and remained stable thereafter, with only minor secondary loss. This CNS graded injury model may be valuable to study the molecular and anatomical mechanisms underlying secondary degeneration and the potency of various posttraumatic treatments in leading to recovery of function.

Aging and brain cholinergic muscarinic receptors: an autoradiographic study in the rat.

Cholinergic muscarinic receptors in aged and young rat brains were studied by quantitative autoradiography of tritiated quinuclidinyl benzilate. A selective pattern of decreased binding density was observed in the aged rat. A large number of regions showed no effect of aging; these include subdivisions of the hippocampal formation and most thalamic and hypothalamic nuclei. Small but significant decreases were found in cortical regions and in the striatum. The largest effects were seen in ventral forebrain cholinergic nuclei, where 40-60% depletions were found in the diagonal band, nucleus basalis magnocellularis, ventral pallidum, and substantia innominata.