Histopathological features of complete pathological response predict recurrence-free survival following neoadjuvant targeted therapy for metastatic melanoma.

BACKGROUND: Recent clinical trials demonstrated the safety and efficacy of neoadjuvant dabrafenib and trametinib (DT) among patients with surgically resectable clinical stage III BRAFV600E/K mutant melanoma. Although patients achieving a complete pathological response (pCR) exhibited superior recurrence-free survival (RFS) versus those who did not, 30% of pCR patients relapsed. We sought to identify whether histopathological features of the pathological response further delineated risk of relapse. METHODS: Surgical resection specimens from DT-treated patients in two phase 2 clinical trials were reviewed. Histopathological features, including relative amounts of viable tumour, necrosis, melanosis, and fibrosis (hyalinized or immature/proliferative) were assessed for associations with patient outcomes. RESULTS: Fifty-nine patients underwent surgical resection following neoadjuvant DT. Patients achieving pCR (49%) had longer RFS compared with patients who did not (P = 0.005). Patients whose treated tumour showed any hyalinized fibrosis had longer RFS versus those without (P = 0.014), whereas necrosis (P = 0.012) and/or immature/proliferative fibrosis (P = 0.026) correlated with shorter RFS. Multivariable analyses showed absence of pCR or presence of immature fibrosis independently predicted shorter RFS. Among pCR patients, mature/hyalinized-type fibrosis correlated with improved RFS (P = 0.035). CONCLUSIONS: The extent and composition of the pathological response following neoadjuvant DT in BRAFV600E/K mutant melanoma correlates with RFS, including pCR patients. These findings support the need for detailed histological analysis of specimens collected after neoadjuvant therapy.

One-Pot Synthesis of Magnesium Aminoclay-Titanium Dioxide Nanocomposites for Improved Photocatalytic Performance.

TiO2 nanoparticles (NPs) with their excellent photocatalytic performance are among the hottest research subjects for environmental-cleanup applications. In the present work, we developed a method of one-pot synthesis of magnesium aminoclay-titanium dioxide [MgAC-TiO2] nanocomposites in ethanol solution and then treated the obtained nanocomposites in a 350 °C muffle furnace for 3 hours. The obtained X-ray diffraction (XRD) patterns confirmed the growth of the anatase TiO2 NPs in the amorphous MgAC phase. In the scanning electron microscopy (SEM) morphological observation, the MgAC-TiO2 nanocomposites exhibited an aggregate form of 246.59 ± 54.20 nm diameter. The synthesis condition entailing loading of 0.3 g MgAC and 5 mL titanium butoxide (TB) (denoted as MgAC [0.3 g]-TiO2 in 40 mL ethanol solution displayed the largest BET surface area, 234.91 m2/g, as well as the largest pore size and pore volume, 6.7131 nm and 0.3942 cm3/g, respectively. Also, MgAC [0.3 g]-TiO2 showed the best photocatalytic performance for methylene blue (MB) on the batch scale under 365 nm wavelength irradiation: a degradation constant rate of 0.0293 min-1, which was ~20-times-better photocatalytic activity than commercial P25. On the pilot scale (100 L), the MgAC [0.3 g]-TiO2 nanocomposite took only ~12 hours to degrade almost MB at 10 ppm concentration. The mechanism of this high photocatalytic activity was determined to be the high rate of adsorption of both MgAC and oxygen vacancies in the anatase phase coupled with the retardation of the rate of recombination of electrons and holes in the TiO2 NPs, the latter proved by photoluminescent quenching tests.

Optimization of spray drying process for Bacillus thuringiensis fermented wastewater and wastewater sludge.

Response surface methodology was used to optimize spray drying process for producing biopesticide powders of Bacillus thuringiensis by using fermented broth of starch industry wastewater and wastewater sludge. Analysis of variance was carried out using number of viable spores in the powder as dependent variable. The determination coefficients of models were 92 and 94% for fermented broth of starch industry wastewater and wastewater sludge, respectively. Under the optimal conditions of the operational parameters of spray drying, the numbers of viable spores were 2.2 × 10(8) and 1.3 × 10(8) CFU/mg in the dry powders for starch industry wastewater and wastewater sludge respectively, with a loss of viable spores of 18 and 13% when compared with their respective fermented broths. The entomotoxicity (measured by the bioassay method) of the powders obtained under optimal conditions showed a loss of 28 and 18% when compared with the fermented broth of starch industry wastewater and wastewater sludge, respectively. The optimized results of spray drying were used for field application calculations. The volume of fermented broth required to produce powder formulated product when compared with the volume required for liquid formulation product in order to treat 1 ha of balsam fir was less and offered several advantages.

Very low loss reactively ion etched Tellurium Dioxide planar rib waveguides for linear and non-linear optics.

We report on the fabrication and optical properties of the first very low loss nonlinear Tellurite planar rib waveguides ever demonstrated. A new reactive ion etch process based on Hydrogen as the active species was developed to accomplish the low propagation losses. Optical losses below approximately 0.05 dB/cm in most of the NIR spectrum and approximately 0.10 dB/cm at 1550 nm have been achieved - the lowest ever reported by more than an order of magnitude and clearly suitable for planar integrated devices. We demonstrate strong spectral broadening of 0.6 ps pulses in waveguides fabricated from pure TeO(2), in good agreement with simulations.

Adaptive pulse shape control in a diode-seeded nanosecond fiber MOPA system.

We demonstrate active shaping of the driving electrical pulses to a laser diode in order to compensate for the pulse shaping effects of gain saturation in an Yb doped fiber amplifier cascade and to allow the generation of user defined customized output pulse shapes. In particular we demonstrate the generation of square output pulses, which have the potential to significantly increase the maximum pulse energy extractable from an amplifier before the peak power reaches the threshold for SRS, and for high efficiency frequency conversion.

Optimisation of cascaded Yb fiber amplifier chains using numerical-modelling.

We show that it is possible to adapt existing software packages developed originally for modeling telecommunication devices and systems to reliably predict and optimize the performance of high-power Ytterbium-doped fiber amplifier and laser systems. The ready availability of a flexible, user-friendly design tool should be of considerable practical interest to scientists and engineers working with this important new laser technology since Ytterbium amplifier and amplifier cascades are often difficult to optimize experimentally due to the three-level nature of the Ytterbium laser transition. As examples of the utility and accuracy of the software, as well as the complexity of the systems and amplifier properties that can be successfully modeled, we present a comparison of experimental and theoretical results for individual core and cladding pumped amplifiers, and also for an ultra-short pulse four-stage amplifier system optimized both to provide a broad gain bandwidth and to minimize nonlinear effects. We also show how high energy 100 ns pulses with complex user definable temporal profiles can be created in a gain-saturated amplifier by suitable pre-shaping of the low-energy input pulses. Furthermore, with appropriate modifications the same software package can be applied to fiber amplifiers based on other rare-earth elements and glass hosts.

Enhancer of rudimentaryp1, e(r)p1, a highly conserved enhancer of the rudimentary gene.

A hybrid dysgenesis-induced mutation, enhancer of rudimentaryp1 (e(r)p1), is a recessive enhancer of a weak rudimentary mutant phenotype in Drosophila melanogaster. The e(r) gene was cloned using P element tagging and localized to region 8B on the X chromosome. It encodes a 1.0-kb and a 1.2-kb transcript. The 1.0-kb transcript is present in both adult males and females, while the 1.2-kb transcript is predominantly found in females. The difference in the lengths of the two e(r) transcripts is caused by two different polyadenylation sites spaced 228 bp apart. The amounts of both of these transcripts are drastically reduced in the e(r)p1 mutant. The P element in e(r)p1 is inserted in the 5'-untranslated leader region near the start of transcription. It may be producing its effect by suppressing transcription and/or by providing transcription termination and polyadenylation signals. The putative e(r) protein is 104 amino acids in length and bears no striking resemblance to protein sequences in GenBank or PIR. While its biochemical function is unknown at this time, sequence analysis indicates that the e(r) protein is highly conserved and, presumably, functionally very important. The amino acid sequences of the D. melanogaster and the Drosophila virilis proteins are 95% identical.

Comparison of llama VH sequences from conventional and heavy chain antibodies.

Forty different PCR clones encoding a llama variable heavy chain domain were analysed. The majority of these clones are derived from heavy-chain antibody cDNA in which the entire CH1 exon is absent. It appears from the amino acid within the VHH framework 1 and 3 that all the llama clones belong to the VH III family. However, the individual llama VHH sequences differ more substantially from each other than expected for members of the same family. Several remarkable amino acid substitutions in the framework 2 hinder the proper association of the VL. However, they lay the foundation for the secretion from the endoplasmic reticulum and good solubility behaviour of llama H2 antibodies. The repertoire of the llama VHHs may be extensive due to the presence of a long CDR3-loop, often constrained by a disulfide bridge and the occurrence of H1 and H2 loop conformations not yet encountered in mice or human VHs. The variability plot of the amino acids in the VHH shows that the first hypervariable region coincides with the structural H1 loop in contrast to the situation found in mice and man where the CDR1 and H1 are slightly offset. We propose that the amino acids of the llama H1 loop participate actively in the antigen binding. All these observations are characteristic for the llama VHHs of the homodimeric heavy-chain H2 antibodies, but are not maintained in the llama clones from conventional heterotetrameric H2L2 immunoglobulins.

The putative cell cycle gene, enhancer of rudimentary, encodes a highly conserved protein found in plants and animals.

The enhancer of rudimentary gene, e(r), in Drosophila melanogaster encodes a protein, ER, whose function has been implicated in pyrimidine biosynthesis and the cell cycle (Wojcik et al. (1994) Genetics 138, 1163-1170). In order to identify conserved regions of the protein and potentially important functional domains, the e(r) gene was cloned and sequenced from two other insects (Drosophila virilis and Aedes aegypti) and three vertebrates (Homo sapiens, Mus musculus, and Brachydanio rerio) and sequenced from a flowering plant (Arabidopsis thaliana). These sequences along with those of a nematode (Caenorhabditis elegans) exhibit a high degree of identity. ER of Drosophila melanogaster is 76% identical to the three vertebrate proteins, 49% identical to the nematode protein, and 40% identical to the plant protein. There is high evolutionary conservation among the vertebrates. The mouse and human proteins are identical and differ from that of the zebrafish by a single conservative amino-acid change (valine for isoleucine). A dramatic sequence conservation is seen in the position of the hydrophobic amino acids. Of the 27 positions occupied by hydrophobic amino acids in ER of Drosophila melanogaster, 25 of the corresponding positions in the human protein, 23 of the positions in Caenorhabditis elegans, and 20 of the positions in Arabidopsis thaliana have hydrophobic amino acids. Most of these residues are present in three conserved amphipathic alpha-helices, which are proposed to function in protein-protein interactions. Two phosphorylation sites for casein kinase II (CKII) have also been conserved within the animal groups. Purified ER from Drosophila melanogaster is phosphorylated in vitro by CKII, arguing that these two sites are functional in vivo. A putative shift in the secondary structure of ER caused by the phosphorylation of these sites suggests that CKII may be regulating the activity of the ER in vivo.

Cellular proliferation, estrogen receptor, progesterone receptor, and bcl-2 expression in GnRH agonist-treated uterine leiomyomas.

Gonadotropin-releasing hormone (GnRH) agonists are commonly used in the treatment of uterine leiomyomas, but little is known about their histological and cellular effects on these neoplasms. We examined a cellular proliferation index as determined by the nuclear antigen Ki-67 and proliferating cell nuclear antigen (PCNA) expression, estrogen receptor (ER), and progesterone receptor (PR) expression in 27 leiomyomas from patients treated with the GnRH agonist leuprolide acetate (LA) and compared them with 33 untreated controls. All leiomyomas were removed by myomectomies from premenopausal woman after 2 to 6 months of LA treatment or in the follicular phase of the menstrual cycle in the untreated controls. Histological features examined included cellularity, nuclear atypia, vascular changes (dilated, thickened, or thrombosed vessels), edema, calcification, hemorrhage, necrosis, hyalinization, and mitotic activity. Although no difference was found between GnRH-treated and nontreated groups with respect to most histological features examined, immunohistochemical studies showed a significant decrease in the cellular proliferation index, ER, and PR expression in the LA-treated cases compared with nontreated controls. The cellular proliferation index, ER, and PR expression decreased by 85%, 49%, and 36%, respectively, in the LA-treated group as compared with controls (P < .001). A subset of cases from the LA-treated and nontreated groups were also analyzed with respect to bcl-2 (an inhibitor of apoptosis) expression, and no significant difference between the LA-treated and nontreated groups was observed with both groups showing a strong (> 75% of cells) cytoplasmic staining pattern. Results of this study show that LA treatment of leiomyomas results in a decrease in number of cycling cells.

Timing of onset of contraceptive effectiveness in Norplant implant users. Part I. Changes in cervical mucus.

OBJECTIVE: To provide scientific data regarding the changes in cervical mucus within the first hours to days after Norplant implant insertion and to estimate when the cervical mucus is hostile enough to suggest a contraceptive effect. DESIGN: Multicenter, clinical descriptive study. SETTING: Family planning clinics. PATIENT(S): Forty-two women who were between days 8 and 13 of their menstrual cycle and who had requested Norplant implants were admitted to the study. INTERVENTION(S): Cervical mucus and blood samples were obtained. MAIN OUTCOME MEASURE(S): Cervical mucus scores, sperm penetration distances, and serum levels of progesterone, estradiol, and levonorgestrel. RESULT(S): The median cervical mucus score observed at baseline was 6 ("fair"), indicating that the mucus was already somewhat hostile before insertion of the Norplant implants. The median scores declined to 5 at 12 and 24 hours and continued to decrease through day 7 to 2 ("poor"), a score that is judged as hostile to sperm penetration. Overall, 73% of all subjects had a poor cervical mucus score by 3 days after insertion; at 7 days after insertion, 90% exhibited poor mucus and none had a good score. There was a substantial drop in the overall median distance traveled by the vanguard sperm after 12 hours for each cervical mucus score grouping. The distance traveled decreased rapidly between 12 and 24 hours to < 0.5 cm in subjects with fair and poor mucus, and by day 3, 91% of the subjects exhibited poor sperm penetration. CONCLUSION(S): On the basis of our findings, deterioration of the quality of the cervical mucus and sperm penetration is evident by 24 hours after insertion, although not to a level that would suggest adequate protection until 72 hours after insertion. Therefore, we are confident in recommending that backup methods of contraception (e.g, condoms) need not be used for more than 3 days after insertion, even when the implants are inserted close to ovulation. These findings provide policy makers, clinic managers, and clinicians with important information about how they can improve client access to Norplant implants.

PIP: To provide clinicians with evidence as to when the cervical mucus is hostile enough in new Norplant implant acceptors to indicate a contraceptive effect, 42 women requesting Norplant from clinical sites in Santo Domingo, Dominican Republic, and Baltimore, Maryland (US), in 1994-95 were enrolled in a clinical descriptive study. At baseline, when all women were between days 8 and 13 of their menstrual cycle, the median cervical mucus score was 6 ("fair") out of a possible maximum of 12. This score declined to 5 at 12 and 24 hours and continued to decrease through day 7, when it reached 2--a level judged hostile to sperm penetration. Overall, 73% of women had a "poor" cervical mucus score by day 3 and 90% were in this category by day 7. There were substantial drops in the overall median distance travelled by the vanguard sperm and in the percentage of subjects demonstrating poor sperm penetration after 12 hours for each cervical mucus score grouping. 91% of women had poor sperm penetration by day 3 and 93% by day 7. These findings suggest that backup contraceptive protection for the entire cycle after Norplant insertion--a standard recommendation--is not necessary given the profound effect of levonorgestrel on cervical mucus shortly after insertion, even in the event of possible ovulation.

Determinants of right-left and top-bottom prevalence for two-dimensional spatial compatibility.

When stimulus and response sets vary along horizontal and vertical dimensions, the horizontal dimension is more dominant than the vertical one, an effect called right-left prevalence. Three accounts have been proposed that attribute the effect to a reduced ability to code vertical locations when horizontal codes are also present, the use of right-left effectors, or a difference in salience of the 2 dimensions. The accounts differ in terms of whether the ability to code and process the 2 dimensions is of limited capacity and whether the prevalence effect is a consequence of the effectors used for responding. The authors report 4 experiments that evaluated these issues. Results indicate that use of right-left effectors is important to the right-left prevalence effect because it increases the salience of the horizontal dimension. However, a top-bottom prevalence effect can be obtained if the vertical dimension is made more salient.

Program directors' perspectives on federally funded fellowship training in primary care research.

PURPOSE: To describe the organization, models of training, and institutional impact of National Research Service Award fellowship programs in primary care research. METHOD: Survey of 25 directors of currently-funded and former training sites. RESULTS: Twenty-four program directors (96%) completed the survey. Programs allocated 39% of fellows' time to course work leading to an advanced degree or other didactic instruction, and 40% of time to the conduct of research. Collaborations with other training programs within the institution occurred at 83% of sites. Programs commonly (54%) or exclusively (42%) relied on a research model of "early research independence" in which the fellow defined an area of research interest, rather than an "apprenticeship" model in which the fellow worked in a senior investigator's research area. These programs enriched the local academic environment, but required extensive financial subsidies. The high costs of training often had adverse impacts on recruitment and other components of the training process. CONCLUSION: Research training programs in primary care often substitute acquisition of advanced degrees for early immersion in research. The "early independence" model of research differs from fellowships in the medical specialties, and requires further study to assess its effectiveness. The need to subsidize training costs poses substantial problems for the institutions that host these fellowship programs.

Determination of arsenic in urine by atomic absorption spectrophotometry for biological monitoring of occupational exposure to arsenic.

An atomic absorption spectrophotometric (AAS) method was successfully applied to analysis of urine for arsenic (As) as a measure of biological monitoring of occupational exposure to As in Vietnam. The application of the method to urine samples from 75 non-exposed control urbanites (after 2-day abstinence from sea foods) gave a reference level of 62.4 +/- 11.6 microg/l (as mean +/- S.D.), from which the upper limit of the normal value (74 microg/l as mean +/- 1 S.D.) and the acceptable limit (100 microg/l as mean +/- 3S.D.) were deduced. Further application to urine samples from 147 workers occupationally exposed to As in Bacthai Non-ferrous Metallurgic Corporation showed significantly elevated levels of As in urine, with mean +/- S.D. of 78.5 +/- 20.2 microg/l. Improvement of working conditions to reduce As exposure resulted in substantial reduction in the ratio of those with urinary As at the level in excess of the acceptable limit. The practical importance of total arsenic determination in urine after 2-day sea food abstinence is discussed in connection with current conditions in analytical laboratories in Vietnam.

Operative laparoscopy for the treatment of ectopic pregnancy in a residency program.

OBJECTIVE: To compare the effectiveness and safety of operative laparoscopy (OL) when used for the treatment of ectopic pregnancy in the setting of a residency training program. STUDY DESIGN: We retrospectively reviewed records of all patients with diagnostic laparoscopy documenting the presence of an ectopic pregnancy. Patients who met the following criteria were selected: hemodynamic stability, tubal distortion no greater than 5 cm and adequate visualization of the pelvis. The cases performed via OL were compared to the cases performed via laparotomy (LAP). All the surgery had been performed by resident physicians with the attending gynecologist as first assistant/supervisor. RESULTS: Ninety-five cases performed from March 1989 to April 1992 met the above criteria. Group 1 consisted of 61 patients treated with LAP. Group 2 consisted of 34 patients treated with OL. The operating time (76.8 +/- 19.0 vs 76.0 +/- 23.0 min, mean +/- SD) and the estimated blood loss (275.0 +/- 183.7 vs. 70.5 +/- 75.2 ml) were similar for both groups. The hospital stay for the LAP group was longer than for the OL group (3.7 +/- 1.2 vs. 1.4 +/- 0.6 days, P < .001). The surgical complication rate was also higher in the LAP group (23% vs 2.9%, P = .16). CONCLUSION: In our residency program the introduction of OL for the management of ectopic pregnancy has resulted in shorter hospital stays and lower morbidity without increased duration of the surgical episode or increased blood loss.

Pregnancy after failed tubal sterilization.

OBJECTIVE: To review the pregnancy outcome in patients with failed tubal sterilization treated in our hospital and to compare the incidence of ectopic pregnancy in our study group with the experience at other military hospitals and with the incidence of ectopic pregnancy in similar patients reported in the literature. STUDY DESIGN: A registry of failed tubal sterilization was started at our hospital in 1989. We entered all information regarding the type of procedure and pregnancy outcome as the patients presented for care. Information on the experience at other military hospitals was obtained through review of data provided by the epidemiology section of the Health Services Command, San Antonio, Texas. RESULTS: From 1989 to 1991, 12 patients with failed tubal sterilization received care at our hospital. Eight patients (67%) had ectopic pregnancies. Of 137 patients identified through the Health Services Command, 49 (36%) had ectopic pregnancies. CONCLUSION: The literature reports a 5-90% incidence of ectopic pregnancy after failed tubal sterilization. The data from our registry and from the Health Services Command corroborate the increased likelihood of ectopic pregnancy in patients with failed tubal sterilization and underscore the need for close follow-up of these patients until the location of the pregnancy is documented.

PIP: Various research studies have documented post-sterilization ectopic pregnancy rates ranging from 5% to 90%. To investigate this complication further, the pregnancy outcomes of the 12 women with failed tubal sterilizations who received care during 1989-91 at the Beaumont Army Medical Center in El Paso, Texas, and of 137 such patients at other Texas military hospitals during 1984-88 were reviewed. Ectopic pregnancies occurred in 8 (67%) women in the former group and 49 (36%) in the latter. The incidence of ectopic pregnancy was similar for minilaparotomy, cesarean section-associated sterilization, and laparoscopy, indicating that there is no significant advantage associated with any one procedure. A post-sterilization pregnancy may be due to tubal recanalization or to a proximal fistula. Recommended is the practice of electrocoagulating both cornual regions to cause scarring of the myometrium and closing of any fistula.

Vesicouterine fistula after cesarean section. A case report.

A 27-year-old woman, gravida 3, para 2002, underwent repeat cesarean section and inadvertent cystotomy, with subsequent development of a vesicouterine fistula. The fistula did not resolve with prolonged bladder catheterization and required total abdominal hysterectomy with resection of the fistula tract and primary closure for definitive treatment.

An unusual cutaneous reaction to anticoagulant therapy.

Unusual complications of warfarin therapy include cutaneous necrosis and the "purple toe syndrome." The latter is more common in men and is not associated with vascular compromise; it usually occurs 3 to 8 weeks after warfarin therapy is begun and may persist for many months after the medication is discontinued. The following is a case of a 63-year-old woman who received warfarin therapy in conjunction with heparin for treatment of a left leg deep vein thrombosis. Approximately 8 hours after receiving her first dose of warfarin (15 mg), she developed acute pain, edema, and discoloration of the entire left leg to the mid-thigh, most prominent in the left great toe. After discontinuation of warfarin therapy, her symptoms completely resolved within 48 hours. This may be a report of a new cutaneous lesion associated with anticoagulant therapy.

Effect of antimicrobial coatings on the radiosensitization of Escherichia coli, Salmonella typhimurium, and Listeria monocytogenes in fresh broccoli.

Radiosensitization of Listeria monocytogenes, Escherichia coli, Salmonella Typhimurium, and aerobic microflora was evaluated in broccoli florets coated by antimicrobial coatings and treated with different doses of γ-radiation. Broccoli florets were inoculated with aerobic microflora isolated from broccoli and with pathogenic bacteria (L. monocytogenes, E. coli, and Salmonella Typhimurium) at 10(6) CFU/ml. Inoculated florets were then coated with methylcellulose-based coating containing various mixtures of antimicrobial agents: organic acids plus lactic acid bacteria (LAB metabolites), organic acids plus citrus extract, organic acids plus citrus extract plus spice mixture, and organic acids plus rosemary extract. Coated florets were irradiated with various doses (0 to 3.3 kGy), and microbial analyses were conducted to calculate the D(10)-value and relative sensitivity. All antimicrobial coatings had almost the same effect of increasing the radiosensitivity of L. monocytogenes (from 1.31 to 1.45 times) to γ-irradiation. The coating containing organic acid plus citrus extract was the most efficient formulation for increasing the radiosensitization of E. coli and aerobic microflora, by 2.40 and 1.76 times, respectively, compared with the control without the antimicrobial coating. The coating containing organic acids plus LAB metabolites was the most effective formulation for increasing the radiosensitization (by 2.4 times) of Salmonella Typhimurium. Results suggest that the spice extract, when mixed with organic acids and citrus extract, might protect E. coli and aerobic microflora from the effects of γ-irradiation.