Correcting transmission losses in short-wave infrared spatially offset Raman spectroscopy measurements to enable reduced fluorescence through-barrier detection.

Spatially offset Raman spectroscopy (SORS) is a proven technique for sub-surface detection. SORS is able to separate Raman signals from a container and its contents, thereby demonstrating application to through-barrier detection for defence and security. Whilst SORS has been demonstrated to reduce fluorescence from the barrier (or surface), fluorescence from the sample (or sub-surface) can still be problematic for some materials when using Raman excitation wavelengths typical in commercially available instrumentation (e.g. 785 nm). Previous work has demonstrated that short-wave infrared (SWIR) excited SORS (e.g. 1064 nm) can reduce fluorescence from the sample and barrier, thereby providing the potential to detect a wider range of materials through a wider range of barriers. In this paper we highlight an additional challenge for detection through some plastic container materials (e.g. high density polyethylene (HDPE) and other opaque plastics) that absorb and scatter both incident and Raman scattered photons in the SWIR band, leading to distortion of the resultant SORS spectrum. The existence of this effect and its impact is explored, along with a potential solution to overcome this challenge that uses multi-wavelength Raman excitation to avoid the detrimental HDPE absorption region.

Laser induced SERS switching using plasmonic heating of PNIPAM coated HGNs.

This is the first report of SERS switching 'on and off' using laser induced plasmonic heating of poly(N-isopropylacrylamide) (PNIPAM) coated hollow gold nanoshells (HGNs). The degree of Raman enhancement for these thermosensitive SERS nanotags was controlled by plasmonic tuning of the properties of the HGNs.

1064 nm SERS of NIR active hollow gold nanotags.

Surface enhanced Raman scattering (SERS) tags are in situ probes that can provide sensitive and selective probes for optical analysis in biological materials. Engineering tags for use in the near infrared (NIR) region is of particular interest since there is an uncongested spectral window for optical analysis due to the low background absorption and scattering from many molecules. An improved synthesis has resulted in the formation of hollow gold nanoshells (HGNs) with a localised surface plasmon resonance (LSPR) between 800 and 900 nm which provide effective SERS when excited at 1064 nm. Seven Raman reporters containing aromatic amine or thiol attachment groups were investigated. All were effective but 1,2-bis(4-pyridyl)ethylene (BPE) and 4,4-azopyridine (AZPY) provided the largest enhancement. At approximately monolayer coverage, these two reporters appear to pack with the main axis of the molecule perpendicular or nearly perpendicular to the surface giving strong SERS and thus providing effective 1064 nm gold SERS nanotags.

Safety and pharmacokinetics of paclitaxel and the oral mTOR inhibitor everolimus in advanced solid tumours.

Everolimus displays antiproliferative effects on cancer cells, yields antiangiogenic activity in established tumours, and shows synergistic activity with paclitaxel in preclinical models. This study assessed the safety and the pharmacokinetic interactions of everolimus and paclitaxel in patients with advanced malignancies. Everolimus was dose escalated from 15 to 30 mg and administered with paclitaxel 80 mg m(-2) on days 1, 8, and 15 every 28 days. Safety was assessed weekly, and dose-limiting toxicity (DLT) was evaluated in cycle 1. A total of 16 patients (median age 54.5 years, range 33-69) were entered; 11 had prior taxane therapy for breast (n=5), ovarian (n=3), and vaginal cancer (n=1) or angiosarcoma (n=2). Grade 3 neutropenia in six patients met the criteria for DLT in two patients receiving everolimus 30 mg weekly. Other drug-related grade 3 toxicities were leucopenia, anaemia, thrombocytopenia, stomatitis, asthenia, and increased liver enzymes. Tumour stabilisation reported in 11 patients exceeded 6 months in 2 patients with breast cancer. Everolimus showed an acceptable safety profile at the dose of 30 mg when combined with weekly paclitaxel 80 mg m(-2), warranting further clinical investigation.

Local inflammation and devascularization--in vivo mechanisms of the "bystander effect" in VPC-mediated HSV-Tk/GCV gene therapy for human malignant glioma.

Somatic gene therapy with the herpes simplex virus type I thymidine kinase gene/ganciclovir (HSV-Tk/GCV) system and murine retroviral vector producer cells (VPCs) was introduced as a new adjuvant treatment modality to treat tumor bulk and to prevent tumor recurrence in patients harboring malignant glioma. The single-center experience after treatment of 27 patients undergoing tumor resection followed by intracerebral VPC injection for HSV-Tk suicide gene therapy will be presented focused on findings of systematic and close MRI follow-up and a few histological specimens. The data indicate that hemorrhagic necrosis due to endothelial cell transfection mediated vessel necrosis and that local inflammatory immune response occurs frequently after gene therapy. These phenomena seem to be specific because none of the patients of a control group showed any similar features. The prognosis (time to progression, survival) of the patients with "bystander effects" after gene therapy was better, but compared to those patients without bystander effects, they were also privileged by a favorable constellation of prognostic factors. Therefore, the appearance of these neuroradiologic features cannot serve as an indicator for treatment effectiveness and outcome.

The role of endogenous opioids in the ovulatory LH surge in mares.

Removal of opioid inhibition of GnRH neurones is thought to be a critical event in generating the ovulatory surge in some species. In the present study, a nonsurgical technique was used to collect pituitary venous blood samples from eight mares every 0.5-1.0 min for 1 h before and after administration of the opioid receptor antagonist naloxone (0.2 mg kg(-1), i.v.), to investigate whether opioid inhibition is also important in mares. Jugular blood samples were taken at 10-15 min intervals. Mares were studied 0, 1 or 2 days before ovulation. Naloxone administration increased mean rates of GnRH (P < 0.01), LH (P < 0.001) and FSH (P < 0.001) secretion. The size of the increment did not vary with proximity to ovulation for any hormone. The amplitude of GnRH pulses rose after naloxone administration (P < 0.05) and the frequency and amplitude of LH pulses increased (frequency, P < 0.05; amplitude, P < 0.02), as did FSH pulse frequency (P < 0.001). Jugular LH and FSH concentrations tended to rise after naloxone administration; however, these changes were not significant. It is concluded that endogenous opioids inhibit GnRH secretion during the period of increasing LH concentration in the ovulatory surge, thereby slowing its rate of increase. It is postulated that treatment with opioid antagonists could be a physiological and non-antigenic way to accelerate and amplify the ovulatory surge in the breeding season. Although a single injection of naloxone is inadequate to do this, it is likely that continuing antagonism, for example with a long-acting, orally-active analogue such as naltrexone, would maintain increased GnRH and LH secretion for sufficient time to raise peripheral LH concentrations and decrease the time until ovulation.

A detailed study of hormonal profiles in mares at luteolysis.

Jugular blood samples were collected at 4 h intervals from six mares during an oestrous cycle to study the hormonal events that occur around the time of luteolysis. Blood samples from day 10 (day 0 = ovulation) until day 3 of oestrus were assayed for prostaglandin metabolite 13,14-dihydro-15-keto PGF2alpha (PGFM), oxytocin, prolactin, progesterone and oestrogen conjugates. PGF2alpha (0.5 or 1.5 mg) was administered to six mid-dioestrous mares and the oxytocin and prolactin responses were measured. One to five large (peak > or =2 x nadir) pulses of PGFM, oxytocin and prolactin were detected in mares during the 3 day period starting on day 13 +/- 0.5. The first PGFM pulse was preceded or accompanied by one or more oxytocin pulses and, overall, large PGFM and oxytocin pulses occurred coincidentally (P < 0.001). During the period of oxytocin and PGFM pulses, progesterone concentrations decreased (P < 0.001) from mid-dioestrous to oestrous values. The first large prolactin pulse occurred as progesterone concentrations approached the nadir and preceded an increase in oestrogen conjugate concentrations by 1.9 +/- 0.6 days. Both PGF2alpha doses significantly increased prolactin concentrations, whereas only the larger dose increased oxytocin concentrations. It is concluded that in mares: (i) PGFM and oxytocin secretion patterns are consistent with the ruminant model of the initiation of luteolysis, in which pulsatile secretion of oxytocin from the posterior pituitary triggers episodic release of uterine PGF2alpha; and (ii) the timing of large prolactin pulses in relation to progesterone and oestrogen conjugates changes indicates that prolactin is more likely to have a role in follicular maturation than in luteolysis.

Simultaneous recording of pituitary oxytocin secretion and myometrial activity in oestrous mares exposed to various breeding stimuli.

The aim of this study was to investigate the effect of different breeding stimuli on uterine contractility and pituitary oxytocin release in five oestrous mares, in order to design better treatments for mares with defective physical uterine clearance mechanisms. Electrodes and strain gauges were implanted surgically on the uterine myometrium and myometrial activity was monitored with a Grass polygraph. A catheter was placed non-surgically in the intercavernous sinus of each oestrous mare to sample pituitary venous blood and a second catheter was placed in the jugular vein. Continuous sampling was performed for 2 h to determine the baseline value and during sequentially applied stimuli of: (i) stallion call; (ii) visual contact with a stallion; (iii) active teasing; and (iv) artificial insemination. No association was observed between uterine contraction and pituitary oxytocin release episodes during baseline recording. Exposure of the mares to any of the breeding stimuli was associated with rapid onset of myometrial contractions (P < 0.0001). The application of a stimulus significantly increased pituitary oxytocin release (P < 0.02) and contraction duration (P < 0.05), and the response to artificial insemination was greater than the responses to the other treatments (P < 0.05). The onset of oxytocin secretion was generally simultaneous with the increase in uterine contraction, rather than preceding it, and in some instances the responses were dissociated, making a cause and effect relationship less likely. In conclusion, interactions between stallions and mares are important components of the uterine clearance mechanism around the time of breeding. Knowledge of these physiological responses may be useful in designing therapeutic regimens for mares with endometritis due to defective uterine contractions.

A phase 1-2 clinical trial of gene therapy for recurrent glioblastoma multiforme by tumor transduction with the herpes simplex thymidine kinase gene followed by ganciclovir. GLI328 European-Canadian Study Group.

This study has investigated the effects of herpes simplex thymidine kinase gene (HSV-tk) transfer followed by ganciclovir treatment as adjuvant gene therapy to surgical resection in patients with recurrent glioblastoma multiforme (GBM). The study was open and single-arm, and aimed at assessing the feasibility and safety of the technique and indications of antitumor activity. In 48 patients a suspension of retroviral vector-producing cells (VPCs) was administered by intracerebral injection immediately after tumor resection. Intravenous ganciclovir was infused daily 14 to 27 days after surgery. Patients were monitored for adverse events and for life by regular biosafety assaying. Tumor changes were monitored by magnetic resonance imaging (MRI). Reflux during injection was a frequent occurrence but serious adverse events during the treatment period (days 1-27) were few and of a nature not unexpected in this population. One patient experienced transient neurological disorders associated with postganciclovir MRI enhancement. There was no evidence of replication-competent retrovirus in peripheral blood leukocytes or in tissue samples of reresection or autopsy. Vector DNA was shown in the leukocytes of some patients but not in autopsy gonadal samples. The median survival time was 8.6 months, and the 12-month survival rate was 13 of 48 (27%). On MRI studies, tumor recurrence was absent in seven patients for at least 6 months and for at least 12 months in two patients, one of whom remains recurrence free at more than 24 months. Treatment-characteristic images of injection tracks and intracavity hemoglobin were apparent. In conclusion, the gene therapy is feasible and appears to be satisfactorily safe as an adjuvant to the surgical resection of recurrent GBM, but any benefit appears to be marginal. Investigation of the precise effectiveness of this gene therapy requires prospective, controlled studies.

Molecular evaluation of biopsy and autopsy specimens from patients receiving in vivo retroviral gene therapy.

We used the polymerase chain reaction (PCR) to assay for the presence of retroviral vector and replication-competent retrovirus (RCR) in autopsy and biopsy specimens from patients who received inoculations of retroviral vector producer cells (VPCs) into brain tumors or apparently normal tissues surrounding resected tumors. The PCR assays were capable of detecting 1 or more proviral copies of vector or RCR in 500,000 cells. Of 113 patients treated in clinical trials between 1994 and 1997, autopsy specimens were available from 32 patients. Brain tumor biopsies were also available from 24 patients. A total of 346 specimens was analyzed. Vector DNA was detected in 55% of tumor samples and 22% of brain samples obtained from resection margins. In contrast, most of the nonbrain tissues were negative for vector DNA; only low levels (<0.03%) of vector sequence were detected in 6 of 240 (2.5%) nonbrain tissues. Vector DNA was not detected in gonadal tissues from 12 men and 10 women. More importantly, RCR was not detected in any of the 134 biopsy and autopsy tissues tested, including all brain tumor, brain, and gonadal specimens. These results comprise the largest data set on molecular analysis of autopsy specimens from patients receiving retroviral gene therapy and indicate that distribution of retroviral vectors following injection of high doses of VPCs is limited to the site of inoculation.

Daily intake and urinary excretion of genistein and daidzein by infants fed soy- or dairy-based infant formulas.

Our aims were to measure isoflavone intake from soy- and dairy-based infant formulas and breast milk and to assess the ability of infants to digest and absorb soy isoflavones by measuring daily urinary excretion rates. We recruited 29 infants: 4 received soy-based formula and 25 received dairy-based formula. We collected pooled urine samples from 3-5 disposable diapers worn during a 24-h period and developed and validated methods for extracting isoflavones from the diapers. Infants were studied every 1 or 2 wk, starting at 2-6 wk of age and continuing until 16 wk. Only soy-based formulas contained isoflavones in concentrations detectable by HPLC (limits: 0.05 mg/L for liquids and 0.1 mg/kg for solids). Soy-based formulas provided a mean (+/-SEM) daily dose of isoflavones (genistein plus daidzein) of 3.2 +/- 0.2 mg/kg body wt, which remained fairly constant (CV: 12%) regardless of age < or = 16 wk. Isoflavones were measurable in all samples from soy-fed infants, but not in urine from dairy-fed infants. Daily isoflavone excretion rates varied little among infants [range of mean individual values (mg x kg(-1) d(-1)): daidzein, 0.37 +/- 0.03 to 0.58 +/- 0.06; genistein, 0.15 +/- 0.03 to 0.32 +/- 0.04] and did not change with age < or = 16 wk. The mean percentage of the daily intake recovered in the urine of soy-fed infants was 38 +/- 4% for daidzein and 13 +/- 3% for genistein, and remained constant with age. These values are similar to those for adults and indicate that young infants are able to digest, absorb, and excrete genistein and daidzein from soy-based formulas as efficiently as do adults consuming soy products.

Gonadotrophin profiles and dioestrous pulsatile release patterns in mares as determined by collection of jugular blood at 4 h intervals throughout an oestrous cycle.

In mares, dioestrous FSH profiles based on once-a-day sampling are variable; however, the pulsatility of plasma FSH, which has been suggested by limited windows of intensive sampling, may contribute to this variability. Jugular blood from six mares was sampled at 4 h intervals throughout an ovulatory cycle to determine cyclic FSH and LH patterns more accurately and to measure gonadotrophin pulse frequency during dioestrus. Synchronous pulses of FSH and LH occurred regularly in all mares between day 4 and day 12 (ovulation = day 0) with a mean (+/- SEM) frequency of 1.9 +/- 0.1 (FSH) or 1.6 +/- 0.1 (LH) pulses day-1. LH pulse amplitude declined (P < 0.0001) between day 4 and day 10, but FSH pulse amplitude remained large and stable, dipping slightly but not significantly on day 6. Daily mean FSH concentrations exceeded (P < 0.0001) early oestrous values between day 4 and day 5, and between day 7 and day 10. However, significantly different patterns were obtained when once-a-day sampling was simulated by selecting samples collected at 08:00 h or noon. LH was higher during the periovulatory surge than during dioestrus (P < 0.0001) and profiles were similar whether daily means or selected samples were used. It is concluded that: (1) the marked pulsatility of plasma FSH during dioestrus makes once-a-day sampling misleading for determining FSH profiles; (2) the dioestrous pattern of large, slow FSH pulses was consistent among mares, unlike that of the daily mean FSH profiles; and (3) no discrete FSH 'surges' were observed during dioestrus, although FSH pulse amplitude tended to undergo alternate increases and decreases. A period of higher amplitude FSH pulses preceded ovulation by 10.2 +/- 0.7 days, which corresponds to the approximate time the ovulatory follicle emerges. Therefore, it is possible that the signal for follicular recruitment in mares is intermittent excursions of plasma FSH above a threshold value.

Comparison of the microheterogeneity of horse LH and FSH in the pituitary with that secreted into pituitary venous blood at oestrus.

For aqueous extracts of pituitary glands of oestrous mares, luteinizing hormone (LH) profiles were found to be similar to each other and to earlier work after chromatofocussing (CF) and isoelectricfocussing (IEF). After CF, both LH and follicle-stimulating hormone (FSH) in pituitary extracts focussed in multiple peaks in the acidic range, with 86% of LH and 80% of FSH found between pH 4 and 6. By contrast, in pituitary venous plasma, only 18% of the LH focussed in this range, whereas a significantly greater proportion (P less than 0.01) eluted above pH 7 than occurred in pituitary extracts (37% vs 2%, respectively). For pituitary venous FSH, there was only a slight shift in the distribution of isoforms compared with the pituitary extract, with a rise in the percentage of strongly acidic molecules in pituitary venous plasma (pH less than 3.65; 34% vs 16%). These results show that at oestrus, horse LH (which differs from that of other species because it has a heavily sialylated C-terminal extension to the beta-subunit, as does eCG), is much more alkaline when secreted as opposed to when it is stored in the pituitary. The authors of this report suggest that this modification is made after entry into a preferentially released pool of LH. Modulation of the forms of LH and FSH that are secreted may play a role in regulating target tissue responses.

Sandimmun (cyclosporin A): mode of action and clinical results in rheumatoid arthritis.

Cyclosporin A (Sandimmun) has gained wide acceptance by most transplant physicians as the immunosuppressant of choice for preventing rejection of solid organ grafts and graft-versus-host disease. The drug has a specific effect on T-lymphocytes in which it seems to prevent the transcription of genes for several lymphokines. The reduction in IL-2 prevents the clonal expansion of T-lymphocytes and their differentiation into effector T-cells. The reduction in IFN-tau interrupts the feedback mechanism between T-cells and macrophages and the aberrant expression of MHC class II molecules. Through these mechanisms Sandimmun exerts an immunosuppressive and anti-inflammatory effect. Considerable evidence has accumulated to suggest that rheumatoid arthritis (RA) is an auto-immune disease. Activated T-lymphocytes interrelate with macrophages, other inflammatory cells and effector cells in joint tissue, leading to symptoms of inflammation accompanied by joint destruction. Immunosuppressive treatment is already well established in this disease and several trials have already taken place using Sandimmun. A total of 224 patients with RA refractory to conventional disease-modifying drugs have participated in 11 published clinical studies. A review of these studies concludes that Sandimmun is efficacious in controlling inflammatory and functional symptoms, although this improvement is no generally accompanied by reductions in ESR and rheumatoid factor. The frequency of adverse events is comparable to that of other treatments but nephropathy remains the principal factor limiting the use of Sandimmun. Recent evidence suggests that with a strict dosage strategy and good monitoring this problem is controllable and reversible. Further studies are under way to confirm these claims.

Culture's influence in Japanese and American maternal role perception and confidence.

Japanese recognition of the importance of the mother in ensuring survival of the culture group is very old. This antiquity is spelled out dramatically in the ancient Chinese-Japanese ideographs (in Japan called kanji), which remain in use today. The kanji reflect the cultural isolation of Japan during 300 years of insularity preceding the Meiji period, and reveal the deep historical link to even greater antiquity in continental Asia. The historical development of kanji explicitly associates the concept of woman with passiveness, the home, and domesticity (see "woman," "peaceful," "to marry," and the concept "wife" in Figure 1) and attributes to the mother survival of the infant (see "mother" and "milk" in Figure 1). The dependence of this new member of the culture group on its mother also is conveyed in the kanji depiction of the "child" with its arms open and unable to walk. In contrast, diverse European progenitors of the comparatively recent "American" perception of the maternal role contributed an array of maternal role expectations; though often conflicting, each set of expectations was well-suited to the exigencies of the particular immigrant group's circumstances. Given the contrasting historical circumstances, substantial difference between present Japanese and American perceptions of the maternal role, and in the degree of a woman's confidence in taking the maternal role for the first time, could be expected. These hypotheses were tested using questionnaire responses and drawings made by 102 Japanese women and 104 American Caucasian women in the last trimester of their first pregnancy. The findings are examined in relation to achievement dynamics in the context of culture.

Breast-feeding as cultural or personal decision: sources of information and actual success in Japan and the United States.

PIP: The impact of sources of information about breastfeeding used by Japanese and American primiparous women during pregnancy is examined in relation to subsequent maternal behavior and success in breast feeding infants during the 1st week, 1st month, and 3rd month after birth. Culture differences were found in the sources of information used. Japanese mothers used more sources reflecting a formal, impersonal, group-oriented learning context, while American mothers used a greater variety of sources, reflecting both an informal, private, personalized approach and a formal approach. Hospital classes taught by nurses were significantly negatively correlated with success in breastfeeding in both cultures (r=0.78, P0.01). Private consultations with health professionals and reading materials (books, magazines, pamphlets) were found to have little or no correlation with success in breastfeeding. The findings indicate that present sources of information on breastfeeding are not achieving the desired result--assisting women in learning to breast feed their infants successfully. Sources of information need to be reevaluated in terms of content, tenor, and the culture learning context in which information is being disseminated.^ieng

The reciprocal impact of breast-feeding and culture form on maternal behaviour and infant development.

PIP: A comparative study of Japanese and American maternal behavior and infant development from birth to 3 months of age, especially in relation to infant feeding practices. The effect of cultural differences on the initiation and prolongation of breastfeeding is discussed, and consideration given to factors such as alertness of the mother during the birth, postpartum procedures in hospital, family sleeping arrangements and the general style of maternal care. The American women in the study were more successful at initiating and continuing breastfeeding their infants, although all the women (Japanese and American) had intended to breastfed only. The possibility of a postpartum "critical" period, and its effect on infant behavior and development, is discussed.^ieng