Fertility regulating and immunotherapeutic vaccines reaching human trials stage

The progress and current status of vaccines which induce antibodiesagainst human chorionic gonadotrophin (HCG) and luteinizinghormone-releasing hormone (LHRH) are reviewed. Three vaccinesdevised against HCG have undergone phase I clinical trials documentingtheir safety, and reversibility. One of these, the heterospeciesdimer (HSD)-HCG vaccine has also completed phase II efficacytrials in sexually active women of proven fertility. Immunizationwith the vaccine prevents pregnancy, as long as the antibodytitres remain =" BORDER="0">50 ng/ml HCG bioneutralization capacity.There is no disturbance of menstrual regularity and women continueto ovulate normally. The antibody response is predominantlyagainst an epitope in the core part of ß-HCG. Fertilityis regained at titres <35 ng. These observations have laidthe scientific foundations of a birth control vaccine. Researchsuggests the feasibility of making a cost-effective recombinantvaccine. The carriers tetanus toxoid (TT) and diptheria toxoid(DT) can be advantageously replaced by peptide determinantsrecognizing T, not B cells. In addition to optional fertilitycontrol, HCG vaccines may have tumour growth inhibition potentialin lung cancers which produce HCG. The vaccine against LHRHcan be used in both males and females. As it is a structurallyconserved molecule, the same vaccine is applicable to both animalsand humans. Antibodies against LHRH block the generation ofgametes and sex steroids, with the result that the vaccine canbe used for fertility control (domestic pets, prolongation oflactation amenorrhoea); as well as for sex hormone-dependentcancers. Phase I/phase II clinical trials have been conductedwith the LHRH vaccine in advanced metastazing carcinoma of prostatepatients with encouraging results. Bioeffective monoclonal antibodieshave been developed against both LHRH and HCG. These can be`humanized' and produced cost-effectively in bacteria and plants,thus paving the way for passive use of such antibodies for immunotherapyof cancers and fertility control.     

Genomic imprinting: potential function and mechanisms revealed by the Prader-Willi and Angelman syndromes

The Prader-Willi (PWS) and Angelman (AS) syndromes are two clinically distinct syndromes which result from lack of expression of imprinted genes within chromosome 15q11-q13. These two syndromes result from 15q11-q13 deletions, chromosome 15 uniparental disomy (UPD), imprinting centre mutations and, for AS, probable mutations in a single gene. The differential phenotype results from a paternal genetic deficiency in PWS patients and a maternal genetic deficiency in AS patients. Within 15q11-q13, four genes (SNRPN, IPW, ZNF127, FNZ127) and two expressed sequence tags (PAR1 and PAR5) have been found to be expressed only from the paternally inherited chromosome, and therefore all must be considered candidate genes involved in the pathogenesis of PWS. A candidate AS gene (UBE3A) has very recently been identified. The mechanisms of imprinted gene expression are not yet understood, but it is clear that DNA methylation is involved in both somatic cell expression and inheritance of the imprint. The presence of DNA methylation imprints that distinguish the paternally and maternally inherited alleles is a common characteristic of all known imprinted genes which have been studied extensively, including SNRPN and ZNF127. Recently, several PWS and AS patients have been found that have microdeletions in a region upstream of the SNRPN gene referred to as the imprinting centre, or IC. Paternal IC deletions in PWS patients and maternal IC deletions in AS patients result in uniparental DNA methylation and uniparental gene expression at biparentally inherited loci. The IC is a novel genetic element which controls initial resetting of the parental imprint in the germline for all imprinted gene expression over a 1.5-2.5 Mb region within chromosome 15q11-q13.      

Connective tissue massage: a bridge between complementary and orthodox approaches

Connective Tissue Massage (CTM) is a bodywork technique which lies at the boundary between alternative and orthodox approaches. A manual therapy technique, it utilizes connective tissue reflex zones which are then manipulated by a gentle shear force with the aim of reducing pain, restoring balance in the autonomic nervous system and inducing segmental and suprasegmental reflex effects on the visceral and hormonal systems. CTM requires a holistic approach in its assessment in order that the degree of autonomic imbalance is established and that subsequent clinical decision making is guided by the zonal positions. The technique, itself, however, is firmly rooted within orthodox scientific concepts and the zones can be seen, palpated and anatomically explained. The approach to treatment, the scientific rationale and research findings are discussed to justify the claims made for this powerful technique.     

The transient receptor potential channel antagonist SKF96365 is a potent blocker of low-voltage-activated T-type calcium channels

Background and purpose:

{"type":"entrez-protein","attrs":{"text":"SKF96365","term_id":"1156357400","term_text":"SKF96365"}}SKF96365 (SKF), originally identified as a blocker of receptor-mediated calcium entry, is widely used diagnostically, as a blocker of transient receptor potential canonical type (TRPC) channels. While SKF has been used as a tool to define the functional roles of TRPC channels in various cell and tissue types, there are notable overlapping physiological and pathophysiological associations between TRPC channels and low-voltage-activated (LVA) T-type calcium channels. The activity of SKF against T-type Ca channels has not been previously explored, and here we systematically investigated the effects of SKF on recombinant and native voltage-gated Ca channel-mediated currents.

Experimental approach:

Effects of SKF on recombinant Ca channels were studied under whole-cell patch clamp conditions after expression in HEK293 cells. The effect of SKF on cerebellar Purkinje cells (PCs) expressing native T-type Ca channels was also assessed.

Key results:

SKF blocked recombinant Ca channels, representative of each of the three main molecular genetic classes (Ca_V1, Ca_V2 and Ca_V3) at concentrations typically utilized to assay TRPC function (10 µM). Particularly, human Ca_V3.1 T-type Ca channels were more potently inhibited by SKF (IC₅₀∼560 nM) in our experiments than previously reported for similarly expressed TRPC channels. SKF also inhibited native Ca_V3.1 T-type currents in a rat cerebellar PC slice preparation.

Conclusions and implications:

SKF was a potent blocker of LVA T-type Ca channels. We suggest caution in the interpretation of results using SKF alone as a diagnostic agent for TRPC activity in native tissues.     

Oral manifestations of inflammatory bowel disease: a review based on the observation of six cases

Inflammatory bowel disease (IBD) comprises two chronic, tissue‐destructive, clinical entities: Crohn's disease (CD) and ulcerative colitis (UC), both immunologically based. Bowel symptoms are predominant, but extra‐intestinal complications may occur, including involvement of the oral cavity. Oral involvement during IBD includes several types of lesions: the most common are aphthae; uncommon lesions include, among others, pyostomatitis vegetans and granulomatous lesions of CD. Starting with a presentation of six patients with oral manifestations, which were crucial for the final diagnosis of IBD, a review on the subject is presented. Oral involvement in IBD may be previous or simultaneous to the gastrointestinal symptoms. However, in the majority of cases, bowel disease precedes the onset of oral lesions by months or years. In many patients, the intestinal symptoms may be minimal and can go undetected; thus, most authors believe that the bowel must be thoroughly examined in all patients with suspected IBD even in the absence of specific symptoms. Usually, the clinical course of oral lesions is parallel to the activity of IBD; therefore, oral manifestations are a good cutaneous marker of IBD.     

High-intensity focused ultrasound ablation as a bridging therapy for hepatocellular carcinoma patients awaiting liver transplantation

The scarcity of liver grafts in Asia leads to a significant dropout of patients from liver transplant waiting lists, particularly patients with hepatocellular carcinoma and a low model for end-stage liver disease score. In order to reduce dropping out, different bridging therapies are employed. We report the use of high-intensity focused ultrasound ablation as a bridging therapy for a patient with hepatocellular carcinoma of stage two and an extremely low platelet count (20×10⁹/L). The ablation was successful. Blood tests showed that his liver function was similar before and after the treatment. No adhesion was encountered in the liver transplantation performed six months later.     

Hilar biliary strictures after liver transplantation: cholangiography and percutaneous treatment

Nonanastomotic hilar bile duct strictures developed in 16 of 152 patients who underwent liver transplantation. The type of pretransplantation liver disease did not significantly affect the likelihood of hilar stricture formation. Possible causes of hilar biliary strictures include hepatic artery occlusion, ductopenic arteriopathic rejection, and cytomegalovirus infection; however, five of the 16 patients had hilar strictures without these complications. Hilar strictures developed within 3 months after transplantation in 11 of the 16 patients. Strictures began as a slight common hepatic duct irregularity and progressed to mucosal cast formation and later to firm strictures. Fifteen of the 16 patients underwent percutaneous stricture dilation. Of 12 patients who no longer have stents, four have had no stricture recurrence for 12-30 months. Eight patients have had to undergo retransplantation or have died. Percutaneous dilations were most likely to result in patient bile ducts if strictures developed within 3 months after transplantation and in the absence of pretransplantation primary sclerosing cholangitis, ductopenic arteriopathic rejection, cytomegalovirus infection, or hepatic artery thrombosis.     

小檗碱对培养大鼠神经细胞内游离Ca^2＋的影响

以Ｆｕｒａ２／ＡＭ为细胞内钙离子的荧光指示剂，用ＡＲＣＭＭＩＣ阳离子测定系统，直接测定了体外培养的新生大鼠神经细胞内游离钙（［Ｃａ２＋］ｉ）值，并观察了小檗碱（Ｂｅｒ）的影响。结果表明，Ｂｅｒ对神经细胞静息［Ｃａ２＋］ｉ无明显影响，Ｂｅｒ１～１００μｍｏｌ·Ｌ－１能剂量依赖地抑制去甲肾上腺素和Ｈ２Ｏ２引起的［Ｃａ２＋］ｉ升高，其ＩＣ５０分别为３９．９和１７．９μｍｏｌ·Ｌ－１。高剂量Ｂｅｒ（１０～１００μｍｏｌ·Ｌ－１）能抑制高Ｋ＋引起的［Ｃａ２＋］ｉ升高。姐果提示，Ｂｅｒ对去甲肾上腺素，高Ｋ＋及Ｈ２Ｏ２引起的［Ｃａ２＋］ｉ升高的抑制作用可能是其抗脑缺血作用机制之一。     

Signal pathway regulation of interleukin-8-induced actin polymerization in neutrophils

Interleukin-8 (IL-8), a recently described peptide cytokine, is a neutrophil chemoattractant and activator that exerts effects similar to fMLP, yet their receptors and their roles in pathophysiology differ. The effect of IL-8 on the neutrophil cytoskeleton has not been well studied; therefore, we compared and contrasted the effects of IL-8 and fMLP on neutrophil actin conformation and on the signal pathway regulation of actin responses. IL-8 caused a rapid, dose-dependent increase in neutrophil F-actin content within 30 seconds. The maximum increase was twofold. These changes were accompanied by the development of F-actin-rich pseudopods, as noted with fluorescence microscopy and scanning electron microscopy. Selected biochemical inhibitors were used to study the regulation of the IL-8-induced actin changes. Incubation of neutrophils with 2 micrograms/mL pertussis toxin resulted in a 67% inhibition of the IL-8-induced F-actin increase. The protein kinase C (PKC) inhibitors, staurosporine and H7, did not inhibit the increase in F-actin caused by IL-8. IL-8 caused a rapid increase in neutrophil intracellular calcium that could be completely inhibited by the chelating agent 1,2-bis(o-aminophenoxy)ethane-N,N-N',N'-tetraacetic acid (BAPTA). However, BAPTA-treated neutrophils retained the ability to increase F-actin in response to IL-8. Similar results were seen with fMLP, indicating that, similar to fMLP, the IL-8-induced actin response is mediated through pertussis-toxin-sensitive G-proteins but is neither dependent on PKC nor increases in cytosolic calcium. Thus, although IL- 8 and fMLP exert their effects on neutrophils through different receptors, the signal transduction pathways used and the effects on actin conformation and pseudopod formation are similar.