The Potential Use of Novel Plant-Derived Recombinant Human Collagen in Aesthetic Medicine.

SUMMARY: Recombinant human type I collagen, identical in structure and functionality to human type I collagen, was successfully expressed and extracted from genetically modified tobacco plants. Contrarily to tissue extracted protein, rhCollagen is not immunogenic and not allergenic and has an intact triple helix structure showing superior biological functionality. A photocurable rhCollagen was developed by chemically modifying the protein to allow cross-linking under illumination at various wavelengths, maintaining the protein structural and biological functions. The use of the photocurable rhCollagen in aesthetic medicine, especially as a dermal filler and as a bioink for 3D-printed breast implant is discussed in this article.

A BMP/activin A chimera is superior to native BMPs and induces bone repair in nonhuman primates when delivered in a composite matrix.

Bone morphogenetic protein (BMP)/carriers approved for orthopedic procedures achieve efficacy superior or equivalent to autograft bone. However, required supraphysiological BMP concentrations have been associated with potential local and systemic adverse events. Suboptimal BMP/receptor binding and rapid BMP release from approved carriers may contribute to these outcomes. To address these issues and improve efficacy, we engineered chimeras with increased receptor binding by substituting BMP-6 and activin A receptor binding domains into BMP-2 and optimized a carrier for chimera retention and tissue ingrowth. BV-265, a BMP-2/BMP-6/activin A chimera, demonstrated increased binding affinity to BMP receptors, including activin-like kinase-2 (ALK2) critical for bone formation in people. BV-265 increased BMP intracellular signaling, osteogenic activity, and expression of bone-related genes in murine and human cells to a greater extent than BMP-2 and was not inhibited by BMP antagonist noggin or gremlin. BV-265 induced larger ectopic bone nodules in rats compared to BMP-2 and was superior to BMP-2, BMP-2/6, and other chimeras in nonhuman primate bone repair models. A composite matrix (CM) containing calcium-deficient hydroxyapatite granules suspended in a macroporous, fenestrated, polymer mesh-reinforced recombinant human type I collagen matrix demonstrated improved BV-265 retention, minimal inflammation, and enhanced handling. BV-265/CM was efficacious in nonhuman primate bone repair models at concentrations ranging from 1/10 to 1/30 of the BMP-2/absorbable collagen sponge (ACS) concentration approved for clinical use. Initial toxicology studies were negative. These results support evaluations of BV-265/CM as an alternative to BMP-2/ACS in clinical trials for orthopedic conditions requiring augmented healing.

An inverse and independent association between Helicobacter pylori infection and the incidence of shigellosis and other diarrheal diseases.

OBJECTIVES: We examined the association between Helicobacter pylori infection and the incidence of diarrheal diseases. METHODS: In a nested case-control study participants were sampled from cohorts of male Israeli soldiers aged 18-21 years, serving in field units and followed up for diarrheal diseases. Case patients (n = 177) were subjects who visited the base clinic because diarrhea and were positive for Shigella sonnei (n = 66), enterotoxigenic Escherichia coli (ETEC) (n = 31) or negative for bacterial pathogens (n = 80; diarrhea of unknown etiology). Controls (n = 418) were subjects who did not suffer from a diarrheal disease during the follow-up. They were matched to case patients by training unit and period. Serum samples were obtained from participants at the beginning of their field training and were tested for anti-H. pylori immunoglobulin (Ig) G and preexisting Shigella sonnei lipopolysaccharide IgG and IgA antibodies using enzyme-linked immunosorbent assay. RESULTS: The proportion of H. pylori-infected subjects was significantly lower among case patients with infection of unknown etiology (36.3%) than among controls (56.0%) (adjusted odds ratio [OR], 0.43; 95% confidence interval [CI], .24-.77; P = .005). The proportion of H. pylori-infected subjects among case patients with S. sonnei shigellosis was also significantly lower than in the control group: 36.3% versus 56.0%. The association persisted after adjusting for sociodemographic variables and preexisting S. sonnei serum IgA antibodies (adjusted OR, 0.37; 95% CI, .14-.95; P = .03) and IgG antibodies (adjusted OR, 0.38; 95% CI, .14-.99; P = .04). The direction of the association between H. pylori infection and ETEC diarrhea was similar, albeit not statistically significant. CONCLUSIONS: Our findings suggest an active role of H. pylori in protection against diarrheal diseases.

Alternative immunological markers to document successful multiple smallpox revaccinations.

BACKGROUND: Successful smallpox vaccination is traditionally defined by clinical response ("take"). Nevertheless, only 60% of subjects in the 2002 Israeli smallpox revaccination campaign developed clinical take. More sensitive immunological markers are needed to document successful revaccination. We compared the level of vaccinia-specific immune markers among subjects who did or did not develop clinical take following revaccination. METHODS: Forty subjects who participated in 2002 smallpox revaccination campaign and developed clinical take were individually matched for age, sex, and smallpox vaccinations with subjects who did not develop clinical take ("no-take"). Vaccinia immunity markers were examined prior to and 14 days and 2 years after revaccination. RESULTS: Higher levels of total immunoglobulin (Ig) G, IgG1, and neutralizing antibodies (highest dilution of serum that inhibited the cytopathic effect by at least 50% [PRNT50]) were observed in the no-take group before vaccination (166 vs 94.3 ELISAU/mL [P<.05], 53.2 vs 34.5 ELISAU/mL [ P<.05], and 30% vs 19.7% [P<.05], respectively). The mean time since last smallpox vaccination was longer in the take group than in the no-take group. Total IgG, IgG subclasses, avidity index, and PRNT50 levels were higher among "take" than "no-take" volunteers 14 days after revaccination. The no-take group was not inferior to the take group in all vaccinia immune marker levels measured 24 months after vaccination. Moreover, mean interferon-γ secretion and the percentage of serum samples with PRNT50 levels ≥1:32 were significantly higher in the No-take group than in the take group (677.5 vs 282.7 pg/mL [P < .05] and 95% vs 80% [P<.05], respectively). CONCLUSIONS: The overwhelming majority of subjects in the no-take group were successfully revaccinated against smallpox. Under circumstances of emergent smallpox mass immunization, there is no need for revaccination success assessment among individuals who have received multiple previous smallpox vaccinations.

Immunosuppressive treatments reduce long-term immunity to smallpox among patients with breast cancer.

BACKGROUND: Mass vaccination is the principal preventive measure against a smallpox outbreak after an act of bioterrorism. Vaccination of subjects who received immunosuppressive therapies is problematic because of smallpox vaccine reactogenicity. Moreover, long-term immunity to vaccinia might be affected. OBJECTIVE: The objective of the study was to examine the effect of cytotoxic chemotherapy on long-term immunity to vaccinia. METHODS: In a case-control study, 67 patients with breast cancer who received cytotoxic chemotherapy and who were disease free for at least 1 year were matched with healthy controls according to age, sex, and the number of smallpox vaccinations received. Markers of immunity to smallpox were examined. Forty-one patients with breast cancer who did not receive chemotherapy were used to assess the affect of cancer and radiotherapy on immunity to smallpox. RESULTS: Patients with breast cancer who received chemotherapy had lower levels of vaccinia total immunoglobulin G and immunoglobulin G1 (expressed as enzyme-linked immunosorbent assay units per milliliter), neutralizing antibodies, vaccinia:memory B cell ratio (expressed as a percentage), and interferon-gamma level (expressed as picograms per milliliter), compared with healthy control individuals. CONCLUSIONS: Immunity to smallpox is reduced after receipt of chemotherapy for breast cancer. This finding should be considered when planning smallpox vaccination campaigns. The effect of immunosuppressive treatments on persistence of immunity should be tested with respect to additional vaccines or natural infections.

High titer human immunoglobulin as a specific therapy against West Nile virus encephalitis.

West Nile virus (WNV) is a mosquito-borne disease found most commonly in Africa, west Asia, and the Middle East, where up to 40% of the human population possesses antibodies. It is an emerging disease in the United States, since 1999 and has spread all over the US and Canada. The virus is maintained in nature in a mosquito-bird-mosquito cycle (primarily Culex), with human horses and other animals serving as incidental hosts. WN infection in humans is usually asymptomatic or involves flu like illness but can develop to severe meningo-encephlitis, with symptoms including cognitive dysfunctions, muscle weakness, paralysis and even death. Elderly and depressed immunity factors are at greatest risk of developing severe neurological disease. Studies in animal models have enhanced significantly the understanding of the viral and host factors that determine the pathogenesis and outcome of WNV disease. Currently, vaccines are available for animal use but there is no effective antiviral therapy or human vaccine for WNV infection. Passive administration of antibodies produced from selected donors has shown promising results in animal models.

A cluster of ecthyma outbreaks caused by a single clone of invasive and highly infective Streptococcus pyogenes.

BACKGROUND: Ecthyma is an invasive, ulcerated skin infection. Four ecthyma outbreaks occurred in different infantry units in the Israeli Defense Force from October 2004 through February 2005. Morbidity attack rates in the first 3 outbreaks were 89% (49 of 55 soldiers), 73% (32 of 44), and 82% (37 of 45). In the fourth outbreak, in which early intervention (antimicrobial treatment and improvement of hygiene) was applied, the attack rate was 25% (10 of 40 soldiers). In the first outbreak cluster, 4 soldiers experienced poststreptococcal glomerulonephritis, and 5 cases of systemic sequelae were recorded (1 case of severe septic shock, 3 cases of pneumonia, and 1 case of septic olecranon bursitis). METHODS: Streptococcus pyogenes and Staphylococcus aureus were isolated from ecthyma sores, oropharynx, and anterior nares of affected and unaffected soldiers involved in all 4 outbreaks. RESULTS: Although the S. aureus isolates had different genomic profiles, >90% of S. pyogenes isolates were identified as belonging to a single clone, emm type 81, T type 8. Epidemiological investigation revealed that the hygiene levels of the soldiers and their living conditions were probably the most important cause for the difference in attack rates, wound severity, and systemic sequelae found between and within the units. CONCLUSIONS: Our study demonstrates the possible ramifications of the combination of a virulent and highly infective S. pyogenes strain and poor living conditions, and it emphasizes the importance of early intervention in such conditions.

Effective post-exposure protection against lethal orthopoxviruses infection by vaccinia immune globulin involves induction of adaptive immune response.

The therapeutic potential of human vaccinia immunoglobulin (VIG) in orthopoxvirus infection was examined using two mouse models for human poxvirus, based on Ectromelia virus and Vaccinia Western Reserve (WR) respiratory infections. Despite the relatively fast clearance of human VIG from mice circulation, a single VIG injection protected immune-competent mice against both infections. Full protection against lethal Ectromelia virus infection was achieved by VIG injection up to one day post-exposure, and even injection of VIG two or three days post-infection conferred solid protection (60-80%). Nevertheless, VIG failed to protect VACV-WR challenged immune-deficient mice, even though repeated injections prolonged SCID mice survival. These results suggest the involvement of host immunity in protection. VIG provides the initial protective time-window allowing induction of the adaptive response required to achieve complete protection. Additionally, VIG can be administered in conjunction with active Vaccinia-Lister vaccination. Vaccine efficiency is not impaired, providing a non-prohibitive VIG dose is used. Thus, VIG can be used as a prophylactic measure against post-vaccinal complications but could also serve for post-exposure treatment against smallpox.

Using high titer West Nile intravenous immunoglobulin from selected Israeli donors for treatment of West Nile virus infection.

BACKGROUND: West Nile Virus (WNV) is endemic in Israel and a significant level of antibodies is present in the population due to natural exposure. Anecdotal cases suggested that the presence of anti-WNV antibodies in intravenous immunoglobulin (IVIG) from Israeli donors (IVIG-IL) assisted the recovery of patients with severe WNV infection. METHODS: To enhance the therapeutic efficacy of IVIG-IL against WNV infection, OMRIX Biopharmaceuticals, Israel, have developed a strategy for selection of plasma units from a 10% fraction of Israeli blood donors with anti-WNV antibodies. Positive units were processed into pharmaceutical grade WNV IVIG (WNIG). Following inoculation with WNV, mice received i.p. injections of different doses (0.01-8 mg/mouse) of IVIG-IL or WNIG, according to the specific experimental protocol. RESULTS: WNIG was about 10 times more potent (per gr of IgG) than was regular IVIG-IL when tested by ELISA and neutralization assays. In a mouse lethal WNV infection model, prophylactic treatment with WNIG was at least 5-10-fold more potent as compared to treatment with IVIG-IL. Treatment with WNIG during active encephalitis, three or four days following WNV infection, had a significant protective effect. WNIG was also very effective in protecting immunosuppressed mice. Indeed, treatment of dexamethasone-immunosuppressed mice with 0.2 or 1.0 mg WNIG 4 h after virus infection, led to 100% survival. CONCLUSION: IVIG produced from selected plasma donated in WNV endemic regions can be used to produce WNV IVIG with superior activity for therapeutic and prophylactic measures.

Postexposure treatment with doxycycline for the prevention of tick-borne relapsing fever.

BACKGROUND: Tick-borne relapsing fever (TBRF) is an acute febrile illness. In Israel, TBRF is caused by Borrelia persica and is transmitted by Ornithodoros tholozani ticks. We examined the safety and efficacy of postexposure treatment to prevent TBRF. METHODS: In a double-blind, placebo-controlled trial, 93 healthy subjects with suspected tick exposure (52 with signs of tick bites and 41 close contacts--those without signs but with a similar risk of contact with ticks) were randomly assigned to receive either doxycycline (Dexxon, in a dose of 200 mg the first day and then 100 mg per day for four days) or placebo after presumed exposure to TBRF. Cases of TBRF were defined by fever and a positive blood smear. Serologic analysis for cross-reactivity to Borrelia burgdorferi and polymerase chain reaction (PCR) for the borrelia glpQ gene were also performed. RESULTS: After randomization, 47 subjects (26 with signs of tick bites and 21 close contacts) received doxycycline. Forty-six other subjects (26 with signs of tick bites and 20 close contacts) received placebo. All 10 cases of TBRF identified by a positive blood smear were in the placebo group of subjects with signs of a tick bite (P<0.001). These findings suggested a 100 percent efficacy of preemptive treatment (95 percent confidence interval, 46 to 100 percent). PCR for the borrelia glpQ gene was negative at baseline for all subjects and subsequently positive in all subjects with fever and a positive blood smear. Seroconversion was detected in eight of nine cases of TBRF. PCR and serum samples were negative for all of the other subjects tested. No major treatment-associated adverse effects were identified. CONCLUSIONS: Treatment with doxycycline is safe and efficacious in preventing TBRF after suspected exposure to ticks in a high-risk environment. (ClinicalTrials.gov number, NCT00237016 [ClinicalTrials.gov].).

Detection of relapsing fever in human blood samples from Israel using PCR targeting the glycerophosphodiester phosphodiesterase (GlpQ) gene.

Relapsing fever caused by Borrelia persica is an acute tick-borne disease infecting people in the Middle East. A PCR test targeting the glycerophosphodiester phosphodiesterase (GlpQ) gene was used to detect infection in the blood of suspected relapsing fever patients. The assay detected infection in all 19 patients from Israel who were spirochetemic by blood smear examination and in two additional patients with clinical relapsing fever who were negative by smear examination. Patients were positive by PCR of blood only at the febrile stage and not during the incubation period prior to the appearance of clinical symptoms. Of 52 tick-bitten subjects who were tested and followed-up after being bitten by ticks, 10 developed symptoms of relapsing fever and all became positive by PCR following an earlier negative test. Partial sequencing of the 16S rRNA gene supported by phylogenetic analysis indicated that infection was caused by B. persica or a closely related species. A phylogenetic analysis of the GlpQ sequence showed that it was different yet closely related to other relapsing fever Borrelia spp. present in the Old World. The GlpQ PCR was positive also with the relapsing fever spirochetes B. recurrentis and B. crocidure but not with the Lyme disease agent B. burgdorferi DNA. A second modified GlpQ PCR was able to discriminate between probable B. persica and B. recurrentis and B. crocidurae infection. This study describes the first molecular assay for the diagnosis of relapsing fever caused by B. persica.

Long-term immunity in young adults after a single dose of inactivated Hepatitis A vaccines.

We evaluated in a prospective study the immune response of naïve subjects to a single dose of inactivated Hepatitis A vaccine. Ninety-seven percent of the vaccinees sero-converted 1 month after vaccination and 93% were still positive 2 years later. All of the vaccinees had a strong booster response 2 years after the single dose. Avaxim was more immunogenic than Vaqta for the primary dose (p = 0.01 for sero-positivity, p<0.001 for antibody level) but no differences were found after boosting with Avaxim. Performance of intense physical activity during the first month after a single vaccine dose was associated with lower antibody levels (p = 0.004). This study indicates that a single dose of inactivated HAV vaccine elicits protective immune memory for at least 2 years.

Community-based safety, immunogenicity, and transmissibility study of the Shigella sonnei WRSS1 vaccine in Israeli volunteers.

We describe the first community-based evaluation of Shigella sonnei strain WRSS1, a live, oral candidate vaccine attenuated by a 212-bp deletion in the virG (or icsA) plasmid virulence gene. Three single-dose regimens of WRSS1 (5 x 10(3) CFU, 2 x 10(4) CFU, and 4 x 10(5) CFU) were tested with cohorts of 15 adult volunteers. The vaccine was generally well tolerated at the 10(3)- and 10(4)-CFU doses. There were no fevers and there was one report of moderate diarrhea in 30 vaccinees; five additional vaccinees reported mild diarrhea. At the 10(5)-CFU dose, there were two reports of low-grade fevers and four reports of moderate diarrhea. The geometric means for immunoglobulin A (IgA) antibody-secreting cells (ASC) against lipopolysaccharide (LPS) were 30, 75, and 193 ASC per 10(6) peripheral blood mononuclear cells (PBMC) for the 10(3)-, 10(4)-, and 10(5)-CFU doses, respectively. The IgG means were 40, 46, and 135 ASC per 10(6) PBMC, respectively. The 10(4)-CFU dose of WRSS1 gave the best balance of safety and immunogenicity, since all vaccinees had a significant IgA ASC response and 73% had a response of more than 50 ASC. The anti-LPS seroconversion rate (threefold) for IgA was 60% and the IgG rate was 27% for the 10(4)-CFU cohort. Each vaccinee and a cohabitating household contact delivered daily perianal stool swabs for bacteriological culture. WRSS1 colonized vaccinees for a median of 5 days, and one individual excreted WRSS1 intermittently for 23 days. None of the 45 household contacts were colonized with WRSS1 after a cumulative 192 days of cohabitation with colonized vaccinees, suggesting that adventitious vaccine spread was not common in the community setting.

The effect of fermented yogurt on the prevention of diarrhea in a healthy adult population.

BACKGROUND: Probiotic dairy products are increasingly gaining popularity. Although the role of probiotic bacteria in the prevention and treatment of pediatric and antibiotic associated diarrhea is fairly well established, their role in the prevention of adult infectious diarrhea has not been well investigated. METHODS: Five hundred forty-one, young male military recruits were enrolled and randomly assigned to receive a yogurt containing Lactobacillus casei (n = 275) or a nonprobiotic yogurt (n = 266). The incidence and duration of diarrhea were documented and stool samples examined for bacteria and parasites. RESULTS: Five hundred and two participants were eligible for final analysis, 254 receiving probiotic yogurt and 248 in the control group. Seventy-one participants (14.14%) experienced diarrhea during the study period. The incidence of diarrhea in the probiotic group and the control group was 12.2% and 16.1%, respectively (P = .207). The mean duration of diarrhea was 3 +/- 1.95 days in the probiotic group and 2.6 +/- 1.08 days in the control group (P = .276). CONCLUSION: Our study demonstrated a nonsignificant trend for reduction of the incidence of diarrhea among healthy young adults consuming yogurt containing Lactobacillus casei . Further study is needed to evaluate the role of probiotics in adults.

Clinical and immune responses after revaccination of israeli adults with the Lister strain of vaccinia virus.

BACKGROUND: During the winter of 2002-2003, the Israeli health authorities launched a campaign to vaccinate first responders against smallpox. METHODS: In an open study, 159 healthy, preimmunized adults, 24-52 years old, who participated in the campaign were vaccinated with the Lister strain of vaccinia virus by the multipuncture technique. The safety, immunogenicity, and reactogenicity of the vaccine were assessed. RESULTS: Successful vaccination rates were 61% and 56%, on the basis of clinical take and seroconversion, respectively. Adverse events among the vaccinees were minor. Seventy-nine (88%) of the 90 vaccinees with clinical take also seroconverted ( kappa =0.779). The level of preexisting antibodies inversely correlated with the rates of clinical take and seroconversion (P