8 kV, 25 μF and 200 Ω. To visualize intracellular expression of WNV proteins, cells were infected or transfected. Two days later, cells were fixed with acetone–methanol (1:1). Cover slips with fixed cells were dried, rehydrated with phosphate-buffered saline and treated with a polyclonal mouse anti-WNV serum (1:50 dilution) obtained after immunization of mice with a formalin-inactivated whole virus vaccine preparation. Bound antibodies were visualized with fluorescein isocyanate-conjugated anti-mouse immunoglobulin (1:100 dilution; Jackson Research Laboratory). Vero or C6/36 cells grown in 175 cm2

tissue culture flasks were infected with either WNVsyn or WNVwt stock at an MOI of 0.0001. The inoculum was removed after 1 h, and 40 ml of fresh medium was added. At various time points (1, 6, 24, 48, 54, 72 and 96 h) 0.5 ml learn more of medium was removed. The infectious virus titer of WNV containing samples was determined by a TCID50 assay. In brief, serial 10-fold dilutions of virus containing supernatant were inoculated in 96-well microtiter plates seeded with Vero cells. After incubation for 7 days at 37 °C and 5% CO2, the plates were screened under a light microscope for the presence of CPE in individual wells. From the number of

CPE positive wells per dilution step, the TCID50 was calculated according to the Poisson formula by means of an in house calculation software HSP inhibitor program. Viral RNA was extracted from supernatant

containing viral material corresponding Farnesyltransferase to 3 × 107 TCID50 by TRIZOL extraction. RNA was precipitated with ethanol and the RNA pellet was resuspended in 50 μl of nuclease-free water. One μl of RNA was used for cDNA transcription using Superscript III cDNA synthesis Kit (Invitrogen) and primers binding in the 3′ end of the NS5 coding region, the NS2B3 coding region and the 3′ noncoding region. For the generation of inactivated whole virus vaccines, the WNVsyn and WNVwt stocks were amplified on BHK cells to serve as prime/boost antigen in animal studies. The WNVsyn preparation (designated CAg 4) as well as WNVwt preparation (designated CAg 6) was prepared in the same manner. Ten roller bottles of BHK cells were infected with a MOI of 0.0001. For better virus yields pH was adjusted to 7.5 after 1 h of virus adsorption. After 4 days of growth the supernatant was harvested and cleared through a low spin centrifugation step at 2500 rpm. The cleared supernatant was treated with formalin (final concentration 0.005%) for 48 h. Next, 30 ml of the inactivated virus was loaded on 5 ml of a 20% sucrose cushion per centrifugation tube (Beckman, SW28 tubes). After 2 h centrifugation with 104,000 × g the supernatant was discarded and resulting pellets were pooled in Tris buffered saline (TBS). An aliquot of the resulting vaccine preparations was subjected to a safety assay to exclude any possible remaining infectivity.

Briefly, rIL-5 was incubated in flat bottom 96-well plates with 2 × 104 BCL1 cells

(a B cell lymphoma line) per well and incubated for 24 h at 37 °C, 5% CO2. 1 μCi of 3H-thymidine (Hartmann Analytic, Switzerland) was added to each well and the plates incubated for 6 h at 37 °C with 5% CO2. The cells were harvested, washed and the incorporation of thymidine determined by emission-counting with a liquid scintillation counter. Commercial murine IL-5 from R&D systems (cIL-5) was used as a control. To test the neutralizing activity of serum http://www.selleckchem.com/products/pci-32765.html from Qβ-IL-5 vaccinated mice, BCL1 cells (2 × 104 per well) were plated in the presence of 20 ng/ml of rIL-5. Pooled sera from Qβ-IL-5 vaccinated or naive mice was titrated with the cells (starting dilution 1/4, titration steps 1/6). After 24 h, 1 μCi of 3H-thymidine was added to the cells, which were incubated for 12 h. The incorporation

of thymidine was determined by emission-counting with a liquid scintillation counter. Murine eotaxin was expressed as a fusion protein in a vector modified from pET22b. The fusion protein (r-eotaxin) consisted of the mature form of murine eotaxin, a hexa-histidine tag and a cysteine containing linker (GGC) at its C-terminus. Expression of r-eotaxin in E. coli BL21 (DE3) was induced with 1 mM IPTG. The soluble fraction of bacterial lysate containing r-eotaxin was mixed with Ni-NTA agarose (Qiagen) in 300 mM NaCl, 50 mM NaH2PO4, 0.5% tween 20 and 20 mM imidazole (pH 8). After washing away unbound contaminants, r-eotaxin was eluted with 300 mM NaCl, 50 mM NaCl, tween 20 and 250 mM imidazole (pH 8). Semi-purified r-eotaxin was loaded onto a click here SP sepharose column (Amersham) in buffer containing 20 mM Tris, 200 mM NaCl (pH 8). After washing r-eotaxin was eluted with an increasing salt gradient (20 mM Tris, 1 M NaCl, pH 8.0). VLPs derived from the bacteriophage Qβ were expressed see more in E. coli containing a expression plasmid pQ10 and purified as described previously [28]. In order to be coupled to IL-5, Qβ VLPs were first derivatized with 10-fold excess of a heterobifunctional chemical cross-liker, succinimidyl-6-(β-maleimidopropionamido) hexanoate

(SMPH). The unbound SMPH was removed by dialysis against PBS. rIL-5 was reduced for 1 h with an equimolar amount of tri (2-carboxyethyl) phosphine hydrochloride (TCEP) in PBS (pH 8.0). Reduced rIL-5 (80 μM) was incubated for 4 h at 22 °C with 40 μM of SMPH derivatized Qβ (dQβ). The reaction was dialysed 12 h against PBS pH 8.0. A slightly different protocol was used to couple r-eotaxin to Qβ⋅ Qβ VLPs were derivatized with a 2.3-fold molar excess of SMPH. A 1.2–1 molar ratio of TCEP to protein was used to reduce r-eotaxin. Reduced r-eotaxin (20 μM) was incubated for 1 h at room temperature with 24 μM of dQβ. The coupling products (Qβ-IL-5 and Qβ-Eot) were analyzed by SDS-PAGE and Western blot with anti-His and anti-Qβ antibodies. Protein concentration was measured by Bradford.

Their Crizotinib order baseline characteristics are presented in Table 1. Ten (53%) participants undertook the control intervention (exercise using either a treadmill or cycle ergometer as prescribed by the treating physiotherapist) first. The two exercise

interventions were conducted for all participants within a 48 hour period, within 72 hours of discharge. Both exercise modes were delivered by the same physiotherapist in the Physiotherapy Gym of the Adult Cystic Fibrosis Unit at The Prince Charles Hospital in Brisbane, Australia. Exercise heart rate and oxygen saturation data during rest and each exercise intervention are presented in Table 2. During the 15-minute exercise, there was no significant difference in the average heart rate between the gaming console exercise of 144 beats/min (SD 13) and control exercise of 141 beats/min (SD 15), mean difference 3 beats/min (95% CI −3 to 9). However, gaming console exercise induced a significantly higher maximum heart rate, by 9 beats/min (95% CI 3 to 15) and a significantly higher minimum heart rate, by 13 beats/min (95% CI 2 to 24). Average, maximum and minimum oxygen saturation during exercise did not differ significantly

between the groups, with between-group differences of only 1–2% (absolute). Participants thought both exercise modes provided a ‘hard’ workout, rating each on average a score of about 15 on the RPE BMS-777607 mouse scale (Table 3). Energy expenditure at rest and during the 15 minutes of exercise is presented in Table 2. No data were recorded for two participants, one each in both exercise interventions. There were no significant differences between the two exercise modes during the 15 minutes of exercise (1.0 MET, 95% CI −0.3 to 0.5). However, there was a significant difference between the two exercise interventions for the total energy expended in the whole exercise session from (26 kcal, 95% CI 17 to 35), as presented in Table 3. The participants’

perception of the exercise is presented in Table 3. Participants rated the gaming console exercise as significantly more enjoyable on the 10-cm visual analogue scale, mean difference 2.6 cm (95% CI 1.6 to 3.6). Participants did not perceive significantly different fatigue or workload between the two types of exercise. Participants thought both exercise modes were an effective form of exercise, rating each on average a score of about 8 on the visual analogue scale. Similarly, participants thought both exercise modes would be feasible to include as part of their regular exercise regimen, rating each on average a score of about 8 on the visual analogue scale. The amount of dyspnoea also did not differ between the two types of exercise. Exercise involving a gaming console appears to be a feasible mode of aerobic exercise for adults with cystic fibrosis.

, 2010). A study modelling the benefits of Barcelona’s scheme identified likely health and environmental benefits, but did not consider equity impacts (Rojas-Rueda et al., 2011), while an evaluation of Montreal’s scheme found that users were more likely to be young,

well-educated, current cyclists (Fuller et al., 2011). An online customer satisfaction survey of 1297 BCH scheme users, found an overrepresentation of young, white, high-earning men (Transport for London,2010d), however its validity was limited by a 5% response rate (personal communication, 2011). This study uses complete registration data from the first seven months of the BCH scheme to compare the personal and area-level characteristics of users with those of the general population, and to examine the predictors of scheme usage.

Transport for London provided anonymised registration data for all users who registered click here between 30th July 2010 and 23rd February 2011 (the most recent data then available). Registration data comprised each individual’s title; date of registration; initial access type (1-day, 7-day or annual); and postcode of registration debit or credit card. Registration data was linked to the total number of BCH trips made prior to 18th March 2011. Our dataset did not include data on pay-as-you-go ‘casual’ users who, since 3rd December, have been able to use the BCH without registering. We used titles to assign gender as ‘male’, ‘female’, or ‘ambiguous’. As proxies for individual-level data, we used postcodes to assign deprivation, PLX4032 mouse ethnicity Sclareol and mode of commute data at the level of the Lower Super Output Area (LSOA, mean population 1500). We assigned small-area income deprivation using the 2010 English Indices of Deprivation (Department for Communities and Local Government, 2011), and assigned the proportions of ‘non-White British residents’ and ‘adult commuters who normally commute by bicycle’ using the 2001 census (Office for National Statistics, 2001). We used postcode centroids to generate distance to the nearest BCH docking station, and to calculate the number of docking stations within 250 m. Our primary measure of BCH usage was ‘mean number of trips per month

of registration’ among individuals who registered for the scheme, with the denominator calculated to include fractions of months. As a secondary outcome we examined whether registering individuals ever used the scheme. Individuals with missing data for any variable (1.2%) were excluded from analyses. We compared personal and area-level characteristics of registered users with area-level characteristics of two populations: a) residents of Greater London and b) all residents and workers in the BCH ‘Zone’. We defined this Zone as all LSOAs where part or all of the LSOA is within 500 m of a BCH docking station, and identified the home postcodes of workers in this Zone using CommuterFlows data from the 2001 census (Office for National Statistics, 2008).

Recently, a new rotavirus vaccine, ROTAVAC®, based on the 116E rotavirus strain and manufactured by Bharat Biotech International Limited of India, demonstrated efficacy in a pivotal clinical trial in India [10] and [11]. Additional rotavirus vaccines are in various stages of preclinical and clinical development. The parameters for the success of such trials from a regulatory perspective will likely differ from the parameters for policy or vaccine introduction decisions, and thus the various study designs used to evaluate efficacy in these trials

are likely to differ. To properly frame the results of clinical trials FG-4592 manufacturer conducted with new vaccines, we reviewed the available literature on efficacy trials of rotavirus vaccines in low-resource settings in Africa and Asia. While acknowledging the importance of safety in regulatory and policy decisions, we limited this review to efficacy outcomes, and to the currently approved and recommended vaccines (Rotarix®, RotaTeq®). Both Rotarix® and RotaTeq® were already approved by

international regulatory authorities www.selleckchem.com/products/PLX-4032.html when tested in Africa and Asia, and thus those trials were conducted primarily to inform policy. Under the assumption that aspects of study design and population characteristics will influence the point estimates of efficacy obtained, we propose that comparisons of point estimates of efficacy from different trials may be challenging, and should be done with a clear understanding of trial design and the variables

that could influence such comparisons. Table 1 provides a number of factors that are known or hypothesized to influence rotavirus vaccine immunogenicity and/or efficacy, with references and examples from clinical trials. We then used these study design characteristics as a framework for evaluating the efficacy data from the new oral rotavirus vaccine, ROTAVAC® as an example of how to interpret appropriately new efficacy results (Table 2). Concomitant administration of oral poliovirus vaccines (OPV) with oral rotavirus vaccines reduces the immunogenicity of rotavirus vaccines, as measured by serum IgA antibody responses and rotavirus vaccine shedding, when compared about with administration of the two vaccines separated in time by 1–2 weeks (Table 1) [12], [13] and [14]. This lower immunogenicity would be expected to result in no effect, or a reduction in efficacy, against clinical outcomes. In moderate to high resource settings, rotavirus vaccines were administered with inactivated poliovirus vaccines (IPV), or separated from OPV administration by at least 2 weeks. In trials performed to date in low resource settings, most of the children received OPV concomitantly with RVs as shown in Table 2. The exception was the trial of RotaTeq® in Africa, where only 35% of children received OPV with RV.

During pandemic situations, the adjuvants may play a critical role in reducing the dose requirement to induce protective immunity in subjects, thereby allowing more people to be vaccinated with limited supply. In this study, a dose-sparing effect afford by squalene-based adjuvant was evaluated by reducing the vaccine dose ranging from 3 μg to

0.004 μg. All of the formulations attained an adequate immune response, achieved theoretically protective HAI titers against H7N9 in mice, and afford substantial cross-reactive HAI titers against H7N7 viral ATM/ATR mutation strain (Fig. 5A–D). To further address the vaccine potency, we also evaluate the protection efficacy

in animals. As the humoral immune response induced by AddaVAX-adjuvanted H7N9 vaccines have reached plateau level at the doses of 1.5 μg and above (Fig. 5, lanes F, G, L, and M), the protection of mice Sirolimus molecular weight against virus challenge were only investigated at the doses of 0.5 μg or less. Virus challenge result showed that 0.5 μg or lower dose (0.004–0.1 μg) of AddaVAX-adjuvanted H7N9 split vaccine were sufficient to provide 100% protection from death in mice (Fig. 6A). However, the group of mice vaccinated with lower dose of H7N9-AddaVAX split vaccines exhibited an dramatically body weight loss (more than 20% of body weight change) in contrast to the mice group receiving 0.5 μg AddaVAX-H7N9 split vaccine (Fig. 6B). This result is consistent with that the 0.5 μg AddaVAX-H7N9 ADP ribosylation factor split vaccine exhibited significantly

predominant immune response against H7N9 virus compared with lower-dose groups (Fig. 5A and B, lane E vs. lanes A–D). All above evidences indicate the squalene-based adjuvantation is a promising way to prepare for effective H7N9 vaccine for surged demand. Accordingly, we highlight that 0.5 μg AddaVAX-H7N9 split virus vaccine is the optimal formulation relevant to providing potent immune response to cross-reaction with H7N7 virus and better protection of mice against H7N9 challenge. Our results also showed that Al(OH)3 can modestly enhance the H7-subtype antigens immunogenicity to move the dose-response curve to lower antigen concentration and works slightly better with high-dose of whole virus (Fig. 2A, lane H vs. b (p < 0.05) and Fig. 4A, lane E vs. Q (p < 0.05)) while the squalene-based adjuvant shifts the optimum immunogenic dose of H7N9 split vaccine at least 10-fold lower ( Fig. 5) and could be proven experimentally in a mouse model. This phenomenon of squalene-based adjuvant enhancing the immune response of poorly immunogenic split antigen is in line with the observation of previous pre-clinical and clinical studies.

This was achieved by enhancing the solubility of the lipophilic MPTS with the application of FDA approved co-solvents, surfactants and their combinations. The aim of the animal studies was therefore dual as the test not only gave answer

to the in vivo efficacy of the drug candidate Bortezomib mouse but would also answer the question of whether the drug shows a fast enough absorption from an intramuscular injection for combating cyanide intoxication. Materials for the conversion test were potassium cyanide (KCN), formaldehyde, ferric nitrate reagent, monobasic sodium phosphate monohydrate and dibasic sodium phosphate anhydrous (VWR International, Suwanee, GA, USA). Methyl propyl trisulfide (50% purity; water solubility = 0.15 ± 0.003 mg/ml) was purchased from Sigma–Aldrich (St. Louis, Missouri, USA), TS were DAPT purchased from VWR International (Suwanee, GA, USA). Ethanol, PEG 200, PEG 300, PEG 400, PG (VWR International, Suwanee, GA, USA), Cremophor EL, Cremophor RH40, sodium cholate, sodium deoxycholate, polysorbate 80 (Sigma Aldrich, St. Louis, MO, USA) were used as solubilizers. Cyclohexanone (Sigma–Aldrich, St. Louis, MO, USA) was used as solvent for the GC–MS measurements. KCN solutions (1.0 mg/ml and 3.5 mg/ml) were used throughout the animal studies. 250, 100 and 50 μl Hamilton

Luer-lock syringes (VWR International, Suwanee, GA, USA) were used in the animal studies with 27G 1/2 needles for intramuscular and Mephenoxalone 25G 1½ needles (VWR International, Suwanee, GA, USA) for subcutaneous injection. In vitro efficacy of MPTS was determined based on

its ability to convert CN to SCN. The method applied was a spectrophotometric measurement of the formed SCN based on the method of Westley (1981) with minor modifications ( Petrikovics et al., 1995). Briefly, 200 μl of various concentrations of SDs, 200 μl of 10 mM phosphate buffered saline, 200 μl of 250 mM KCN and 400 μl of deionized water were mixed. The reaction was incubated for 5 min and was quenched with 500 μl of 15% (v/v) formaldehyde. 1.5 ml of ferric nitrate reagent was added to form a reddish brown complex (Fe(SCN)3) that was quantitatively determined at 464 nm using a spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA). Tests were performed with MPTS and TS at concentrations ranging from 25 mM to 0.156 mM with two fold serial dilutions in between. The solubility of MPTS was determined in co-solvents, surfactants and their combinations. Aqueous solutions of co-solvents and surfactants were prepared at 10%, 25%, 50%, 75%, 90% and 1%, 5%, 10%, 15%, 20% respectively. Based on the solubility enhancing efficacy of the co-solvent/water and surfactant/water systems the most effective excipients were combined into one system forming a co-solvent/surfactant/water system.

The authors thank and acknowledge the contribution of participation of the infants and parents in Taipei, Taoyuan, Taichung (Taiwan),

as well as the investigational staff at National Taiwan University Hospital, Taipei; Chang Gung Children’s Hospital, Taoyuan; Mackey Memorial Hospital, Taipei; Taichung Veterans General Hospital, Taichung; Far Eastern Memorial Hospital, New Taipei City; and at Sanofi Pasteur: Helena Aurell, Isabelle Bruyere, Murielle Carre, Nicolas Corde, Sophia Gailhardou, Christel selleckchem Guillaume, Julia Lin, Agnes Machmer, Celine Monfredo, Zulaika Naimi, Karen Privat, Camille Salamand, Nuchra Sirisuphmitr. This manuscript was prepared with the assistance of a professional medical writer, Alice Walmesley, and funding from Sanofi Pasteur. “
“Most of the serious morbidity and mortality associated with seasonal influenza occur in people 65 and older [1], [2], [3], [4], [5] and [6]. This increasingly large part of the population is a priority for influenza vaccination, but the current vaccine is less effective in

older than younger adults [7] and [8]. In response to the demand for new vaccines that elicit a stronger immune response in older adults, Selisistat various types of influenza trivalent inactivated vaccines (TIVs) are available [9], [10], [11], [12] and [13]. Influenza vaccine effectiveness (VE) is a major consideration in the choice of vaccine, but the relative effectiveness of TIVs in older adults is not well established. Data from direct comparisons of TIVs are needed to inform decisions about which vaccine to use. To be used during the 2011–2012 season, three vaccines were acquired by public tender by the Valencia Autonomous Community (Valencia region) government, and centrally distributed to be offered free of charge to groups targeted for GBA3 influenza vaccination [14]: a split trivalent classical intramuscular vaccine (Gripavac®; Sanofi-Pasteur MSD, Lyon, France); a virosomal trivalent subunit vaccine (Inflexal-V®, Crucell, Leiden, The Netherlands); and a split trivalent intradermal vaccine (Intanza® 15 μg, Sanofi-Pasteur MSD, Lyon, France). The intradermal

TIV seasonal influenza vaccine delivered by a microneedle injection system (Intanza® 15 μg) and the virosomal TIV, intramuscularly delivered influenza vaccine (Inflexal® V) were targeted free of charge to adults ≥65. Enhanced immune response in the elderly is thought to be achieved differently by each vaccine type. Intradermal vaccination provides direct access to the immune system through the dermis, which is rich in immune cells and highly vascularized with an extensive lymphatic network [11] while virosomal vaccination induces high virus-neutralizing antibody titers and primes the cellular arm of the immune system [15]. Health authorities expressed no preference for either vaccine, and both vaccines were widely distributed [14]. Several sources of data can be used to estimate relative TIV effectiveness in Valencia region.

, 2002, Jabbour et al., 2012, Mckee et al., 2002 and Rechel and Mckee, 2007). For example, in Qatar, the life expectancy at birth is the highest in the world as a result of the lower NCD mortality rate in the Qatari men. This may be attributed to the establishment of its Supreme Council selleck chemical of Health, which has taken positive steps in tackling health inequity by involving government ministries, non-governmental agencies and industries (Jabbour et al., 2012). On the other hand, for some countries in the upper middle income countries, such as Turkmenistan, Kazakhstan

and Russia, the life expectancy remained short at 60, 62 and 63 years, respectively. In Turkmenistan, this has been attributed to the political turmoil where healthcare funding and healthcare workforce

declined resulting in reduced accessibility to health care (Rechel and McKee, 2007). In Kazakhstan and Russia, men’s shorter life expectancy is mainly due to excessive alcohol consumption, heavy smoking, high-fat diets and sedentary lifestyle (Cockerham et al., 2002 and Mckee et al., 2002). For communicable diseases in Asia, the male mortality rate (162.0 deaths per 100,000) is higher than that in Europe (50.9 deaths per 100,000), USA (29.8 deaths per 100,000) and Australia (15.4 deaths per 100,000) (WHO, 2008). Timor-Leste, Myanmar, Cambodia and Afghanistan have the highest mortality rate due to communicable disease for men in Asia (422.3 to 565.4 deaths per 100,000). Among Asian countries, Timor-Leste has the highest male mortality due to tuberculosis Veliparib ic50 and sexual transmitted infections; Myanmar has the highest male mortality rate due to HIV/AIDS; Afghanistan has the highest male mortality rates due to respiratory infection, hepatitis B and hepatitis C; while Cambodia has the second highest male mortality rate in hepatitis

B, hepatitis C and sexual transmitted infections (Tan et al., 2013). The high mortality in these countries is likely to be attributed to poverty and less-than-effective health care system (Gupta and Guin, 2010). This study found that majority of the higher-income countries faced transition toward chronic non-communicable disease while the middle- and low-income countries faced Cediranib (AZD2171) double disease burden of communicable and non-communicable diseases. The male mortality rate due to non-communicable diseases in Asia (759.7 deaths per 100,000) is higher than Europe (616.9 deaths per 100,000), the USA (485.9 deaths per 100,000) and Australia (389.2 deaths per 100,000). Male mortality rate due to injuries is higher compared to female in all Asian countries. Among the highest in Asia are Iraq, Sri Lanka and Afghanistan, where the figures are contributed by war. For Russia and Kazakhstan, the main causes are accidental poisoning by and exposure to noxious substances and other intentional injuries.

2). He authored approximately 280 articles and many book chapters and books, with contributions from across the entire spectrum of cardiac and vascular diseases.

He was active in many professional groups, was a visiting professor and lecturer on cardiac disease worldwide, and served on the editorial boards of several medical journals, including Cardiovascular Pathology, Circulation, American Heart Journal, Human Pathology, and Modern Pathology. check details Dr. Titus was a visiting professor in many medical schools throughout the world and received multiple other honors including the R.T. Hall Lectureship of the Cardiac Society of Australia and New Zealand. Dr. Titus also received a “Service to Humanity” Award in 2004 from the United Hospital Foundation for his “selfless leadership in improving the health and welfare of Saint Paul (MN) and the surrounding communities.” He served as president of the Houston Society of Clinical Pathologists, from which he also received the Harlan Spjut Award for Distinguished Scholarly Achievement in 1993. He was honored in 2006 by the Texas Society of Pathologists with the John J. Andujar Dabrafenib in vitro Citation of Merit. Jack had an enviable knowledge base, impeccable wisdom, and a wonderful and ever-present keen sense of humor, all of which he shared generously. Early in my career, when still a resident in anatomic pathology and seeking a mentor and

case material, I contacted Mannose-binding protein-associated serine protease him and requested the opportunity to spend 3 months at The Methodist Hospital in Houston reviewing specimens and medical records of patients who had had valve replacement on a Cardiovascular Surgery Service led by the famed surgical pioneer and innovator, Dr. Michael DeBakey. I owe Jack

great debt for arranging an unimaginably formative opportunity, during which he introduced me to colleagues, including other leading surgical collaborators, arranged for me to review the autopsy and medical records of approximately 400 valve replacement patients, and spent many hours discussing and providing a highly skilled and thoughtful approach to cases, studies, and results derived from them. This experience was a most important catalyst to my career, and I had the privilege of many professional and other conversations with Jack since those several months working closely together over 30 years ago. I admired him greatly not only for his technical expertise, but also for his warmth, approachability, and strong commitment to family. In each encounter, he never failed to ask, with sincere interest, about the health and accomplishments of my wife and children. Indeed, Jack Titus also had a rich personal and family life. Shortly following his college graduation, he married Beverly J. Harden, in South Bend, his highly supportive and loving wife of 62 years and who now survives him (Fig. 3).