Green synthesis of near-infrared absorbing eugenate capped iron oxide nanoparticles for photothermal application.

Nanomaterials exhibit different interesting physical, chemical, electronic and magnetic properties that can be used in a variety of biomedical applications such as molecular imaging, cancer therapy, biosensing, and targeted drug delivery. Among various types of nanoparticles, super paramagnetic iron oxide nanoparticles (SPIONs) have emerged as exogenous contrast agents for in vitro and in vivo deep tissue imaging. Here, we propose a facile, rapid, non-toxic, and cost-effective single step green synthesis method to fabricate eugenate (4-allyl-2-methoxyphenolate) capped iron oxide nanoparticles (E-capped IONPs). The magnetic E-capped IONPs are first time synthesized using a medicinal aromatic plant, Pimenta dioica. The Pimenta dioica leaf extract was used as a natural reducing agent for E-capped IONPs synthesis. The crystalline structure and size of the synthesized spherical nanoparticles were confirmed using the x-ray diffraction and electron microscopic images respectively. In addition, the presence of the functional groups, responsible for capping and stabilizing the synthesized nanoparticles, were identified by the Fourier transform infra-red spectrum. These nanoparticles were found to be safe for human cervical cancer (HeLa) and human embryonic kidney 293 (HEK 293) cell lines and their safety was established using MTT[3-(4, 5-Dimethylthiazol-2-yl)-2, 5-Diphenyltetrazolium Bromide] assay. These green synthesized E-capped IONPs display a distinct absorbance in the tissue transparent near-infrared (NIR) wavelength region. This property was used for the NIR photothermal application of E-capped IONPs. The results suggest that these E-capped IONPs could be used for deep tissue photothermal therapy along with its application as an exogenous contrast agent in biomedical imaging.

Neurodevelopmental Outcomes in Children Born to Climate Refugee Mothers in Bangladesh: Experiences from Cyclone Aila.

Cyclone Aila hit the South-West coast of Bangladesh in May 2009, when in Dacope Upazilla over 50,000 people were left homeless as climate refugees (CRs) for over two years. We determined neurodevelopmental status of children born as CRs compared to their non-Climate Refugee (NCR) counterparts. Pregnant mothers were enrolled from May 2009 to April 2010 in entire Dacope in a study which profiled their health conditions. From among these mothers, 12 months post-Aila 267 CR mother-child dyads, and 552 NCR mother-child dyads were enrolled to assess their children's neurodevelopmental outcomes. There were significantly more landless families among CRs compared to NCRs (p value = 0.0001; OR = 1.86, 95% CI: 1.37 - 2.51). The mean±SD age at assessment of CR children was 8.52±4.57 months compared to a mean age 9.09±4.13 months of the NCR children (p=0.610). Neurodevelopmental Impairments (NDIs) were three times higher in the former (21.3%), compared to the latter (7.4%) group (p=0.0001; OR 3.83, 95% CI: 2.16 - 5.21). Specifically, expressive language (p value 0.002; OR 2.86, 95% CI: 1.46 - 5.57) and gross motor functions (p=0.007; OR 2.27, 95% CI 1.22 - 4.20) were the most significantly affected areas of impairment. Children born to CR mothers had a three times higher proportion of NDIs. The findings are of concern as in Bangladesh large populations are forced to leave their homes and become CRs annually. Optimum antenatal care of pregnant women as well as their offsprings within refugee situations needs to be ensured to prevent NDIs and poor quality of survival.

Neonatal uterine prolapse - a case report.

Uterine prolapse is commonly seen in the geriatric age group. Congenital vaginouterine prolapse is a rare condition occurring in neonates and is usually associated with spinal cord malformations in about 85% of cases. Several modalities of treatment have been described for neonatal uterine prolapse. Conservative treatment in the form of simple digital reposition, use of pessary or other self-retaining device is usually sufficient to treat this condition, which is self-limiting and regressive. Here we report our first case of neonatal uterine prolapse, managed successfully with simple digital reposition.

Caudal anesthesia in pediatric surgical practice.

Prospective study was carried out on 100 patients since May 2005 in my private practice and in the department of pediatric surgery of MMCH. Under caudal anesthesia along with or without ketaminie induction and gas inhalation all the patients underwent different surgical procedure namely anorectal surgery (eg. anoplasty, rectal polyp), urogenital surgery (Circumcision, hypospadias, meatotomy), groin surgery (hernia, hydrocele) and foot & leg surgery. Calculated dose schedule of drugs used in anesthesia and volume were maintained. Time of giving anesthesia and time of starting analgesia were recorded. Per-operative and postoperative analgesia were evaluated. Every parent was explained regarding the merit of caudal anesthesia calculated and compared with that of general anesthesia. Application of caudal anesthesia with or without ketamine & diazepam induction can be used safely and cost effectively and may be put into protocol in many of the pediatric surgical practice both in institute and also in private practice.

Multi-class classification algorithm for optical diagnosis of oral cancer.

We report development of a direct multi-class spectroscopic diagnostic algorithm for discrimination of high-grade cancerous tissue sites from low-grade as well as precancerous and normal squamous tissue sites of human oral cavity. The algorithm was developed making use of the recently formulated theory of total principal component regression (TPCR). The in vivo autofluorescence spectral data acquired from patients screened for neoplasm of oral cavity at the Government Cancer Hospital, Indore, was used to train and validate the algorithm. The diagnostic algorithm based on TPCR was found to provide satisfactory performance in classifying the tissue sites in four different classes - high-grade squamous cell carcinoma, low-grade squamous cell carcinoma, leukoplakia, and normal squamous tissue. The classification accuracy for these four classes was observed to be approximately 94%, 100%, 100% and 91% for the training data set (based on leave-one-out cross-validation), and was approximately 90%, 90%, 85% and 88%, respectively for the corresponding classes for the independent validation data set.

N2 laser excited autofluorescence spectroscopy of formalin-fixed human breast tissue.

The paper reports results of an in vitro study on autofluorescence spectroscopy of fresh and formalin-fixed human breast tissue samples to investigate the effect of formalin fixation on the measured autofluorescence spectra. It also explores the applicability of the approach in discriminating cancerous from the uninvolved sites of the formalin-fixed breast tissues based on their autofluorescence spectra. A probability-based diagnostic algorithm, making use of the theory of relevance vector machine (RVM), a powerful recent approach for statistical pattern recognition, was developed for that purpose. The algorithm provided sensitivity values of up to 97% and specificity values of up to 100% towards cancer for both the independent validation data set as well as for the training data set based on leave-one-out cross-validation. These results suggest that autofluorescence spectroscopy may prove to be a valuable additional in vitro diagnostic modality in clinical pathology setting for discriminating cancerous tissue sites from normal sites.

Support vector machine for optical diagnosis of cancer.

We report the application of a support vector machine (SVM) for the development of diagnostic algorithms for optical diagnosis of cancer. Both linear and nonlinear SVMs have been investigated for this purpose. We develop a methodology that makes use of SVM for both feature extraction and classification jointly by integrating the newly developed recursive feature elimination (RFE) in the framework of SVM. This leads to significantly improved classification results compared to those obtained when an independent feature extractor such as principal component analysis (PCA) is used. The integrated SVM-RFE approach is also found to outperform the classification results yielded by traditional Fisher's linear discriminant (FLD)-based algorithms. All the algorithms are developed using spectral data acquired in a clinical in vivo laser-induced fluorescence (LIF) spectroscopic study conducted on patients being screened for cancer of the oral cavity and normal volunteers. The best sensitivity and specificity values provided by the nonlinear SVM-RFE algorithm over the data sets investigated are 95 and 96% toward cancer for the training set data based on leave-one-out cross validation and 93 and 97% toward cancer for the independent validation set data. When tested on the spectral data of the uninvolved oral cavity sites from the patients it yielded a specificity of 85%.

Depth-resolved fluorescence measurement in a layered turbid medium by polarized fluorescence spectroscopy.

We show that, when a turbid medium with a layered fluorophore distribution is excited by linearly polarized light, measurement of angle-resolved polarized fluorescence can provide depth-resolved fluorescence measurements.

Fluorescence depolarization in a scattering medium: effect of size parameter of a scatterer.

For a monodisperse scattering medium, we investigate the dependence on scatterer size parameter for the change in anisotropy of fluorescence due to single scattering at excitation or emission wavelength. The value for the ratio of the anisotropy of fluorescence after one scattering at excitation or emission wavelength to the initial value was observed to increase with increasing value of scatterer size parameter. The effect of multiple scattering on anisotropy of fluorescence from fluorophores embedded in a scattering medium was incorporated using a photon migration model. The model was validated by experiments carried out on samples with known concentration of polystyrene microspheres as scatterers and riboflavins or reduced form of nicotinamide adenine dinucleotide as fluorophores.

Polarized fluorescence spectroscopy of human tissues.

We report the results of a study carried out to investigate the effect of blood absorption on polarized and unpolarized fluorescence from resected tissue samples and tissue phantoms. The signatures of blood absorption were found to be significantly smaller in polarized fluorescence than in unpolarized fluorescence spectra. The reduced effect of blood absorption on polarized fluorescence also leads to reduced site-to-site variability in polarized fluorescence intensity and line shape compared with unpolarized fluorescence.

Measurement of optical transport properties of normal and malignant human breast tissue.

We report measurement of optical transport parameters of normal and malignant (ductal carcinoma) human breast tissue. A spatially resolved steady-state diffuse reflectance technique was used for measurement of the reduced scattering coefficient (mu(s)?) and the absorption coefficient (mu(a)) of the tissue. The anisotropy parameter of scattering (g) was estimated by goniophotometric measurements of the scattering phase function. The values of mu(s)? and mu(a) for malignant breast tissue were observed to be larger than those for normal breast tissue over the wavelength region investigated (450-650 nm). Further, by using both the diffuse reflectance and the goniophotometric measurements, we estimated the Mie equivalent average radius of tissue scatterers to be larger in malignant tissue than in normal tissue.

Depolarization of autofluorescence from malignant and normal human breast tissues.

We report on steady-state measurements on the anisotropy of autofluorescence from malignant and normal breast tissue as a function of tissue thickness. For thin tissue sections the anisotropy from normal tissue was found to be smaller compared with that from malignant tissue. However, the opposite result was obtained for thicker tissues. A phenomenological model was also developed to simulate the dependence of anisotropy on tissue thickness.

Breast cancer diagnosis using N2 laser excited autofluorescence spectroscopy.

BACKGROUND AND OBJECTIVE: This article reports results of an in vitro study involving 63 patients for the evaluation of the diagnostic potential of N2 laser excited autofluorescence spectroscopy of human breast tissues. MATERIALS AND METHODS: The N2 laser-excited spectra were recorded from benign (fibroadenomas, 35 patients), cancerous (ductal carcinomas, 28 patients), and normal (the uninvolved areas of the resected cancerous specimens). A stepwise multivariate linear regression (MVLR) analysis was developed to analyze the diagnostic content of the breast tissue fluorescence spectra. RESULTS: Significant changes were observed in the autofluorescence from normal, benign, and cancerous breast tissues, particularly in the spectrally integrated fluorescence intensity. The ratio of mean spectrally integrated intensity from cancerous tissues to that from benign tumor and normal tissues were 3.2 and 2.8, respectively. A discrimination parameter based on spectrally integrated intensity alone provided a sensitivity and specificity of up to 99.6% over the sample size investigated for discrimination of cancerous breast tissues from benign/normal. CONCLUSION: Our results suggest that a straightforward measurement of the total integrated fluorescence intensity can provide excellent discrimination between cancerous and benign/ normal breast tissues.

In vitro activities of PS-15, a new dihydrofolate reductase inhibitor, and its cyclic metabolite against Mycobacterium avium complex.

PS-15, a new dihydrofolate reductase inhibitor, and its cyclic metabolite were evaluated for in vitro activity against 31 clinical Mycobacterium avium complex isolates. Broth dilution MICs of PS-15 ranged from 16 to 64 micrograms/ml. The cyclic metabolite was three to five times more active than the parent compound. Further evaluation of these compounds in an M. avium-infected murine test system will be of interest.

Remote sensing system to study rodent behaviour.

The instrumental system assembled consists of video monitors which projects signals from remotely placed video camera in the rat pen with pan and tilt arrangement and all operations are done through control-room. Signals obtained from the camera will pass through the monitor and then fed into VCR system which records the behavioural events. Studies carried out with the above system has revealed that all the characteristic behaviour of gerbils (Meriones hurricane Jerdon) which is significantly superior (P < 0.001) over the direct observation method. The integrated sensing system is an ideal instrumental set up for viewing and recording the behaviour of rodents as well as other animals in the experimental pen throughout the year under varying weather and light conditions.

Cytotoxicity of certain organic solvents and organophosphorus insecticides to the ciliated protozoan Paramecium caudatum.

Responses of Paramecium caudatum, a ciliated protozoan, to acute exposures of certain organic solvents and organophosphorus insecticides (OPI) were studied by determining their lethal concentration (10 min-LC100) and median lethal concentration (4 h-LC50). The solvents and OPI evoked a distinct sequence of responses. Among the five solvents tested, acetone proved most toxic [LC-2.9% and LC50-0.68% (v/v)], while dimethyl sulphoxide (DMSO) showed least toxicity [LC-11.0% and LC50-3.16% (v/v)]. The order of toxicity of solvents was: acetone greater than ethanol greater than methanol greater than N, N-dimethylformamide greater than dimethylsulphoxide. The LC values of six OPI dissolved in either acetone or DMSO indicated that they were more toxic when dissolved in acetone and least toxic in DMSO. Among the OPI, bromophos proved most toxic (LC-10 ppm) while malathion showed least toxicity (LC-200 ppm) in DMSO. The order of toxicity of OPI was: bromophos greater than pirimiphos-methyl greater than parathion methyl greater than dichlorvos greater than fenitrothion greater than malathion. The 4 h-LC50 values computed for bromophos and malathion (dissolved in DMSO) were 575 ppb and 19.9 ppm, respectively, indicating the high susceptibility of P. caudatum to bromophos. The results indicate that the Paramecium toxicity assay could be used as a complementary system to rapidly elucidate the cytotoxic potential of compounds.

Serum hexachlorocyclohexane residues in workers engaged at a HCH manufacturing plant.

The accumulation of hexachlorocyclohexane, one of the most commonly used chlorinated insecticides, was studied in a high-risk group of people who are exposed during its manufacturing process. The serum HCH concentration was estimated by quantitating its alpha, beta, gamma and delta-isomers with the help of GLC analysis. Exposed workers involved in maintenance work were found to have 3 times higher HCH residues than the controls, while in the plant operators and supervisors the levels were 5 times higher. The most severely exposed were the handlers who are in direct contact with the insecticide. The level of serum HCH residues in them was found to be about 12 times higher than those in the controls. The percentage composition of the insecticide, with respect to the different isomers, varied up to 20% for the gamma-isomer, up to 30% for alpha-isomer and had between 60%-100% of the beta-isomer in the samples. The serum levels of the insecticides were clearly related to the workers' job-related close contact, intensity, frequency, and the duration of their exposure. The total of HCH residues in the sera studied were in the range of 0.143-1.152 ppm. The importance of these findings and the relevant protective measures are discussed.

Olfactory behavior of red flour beetleTribolium castaneum (Herbst) (Coleoptera, Tenebrionidae) towards natural fatty acid esters.

The olfactory responses of red flour beetles,Tribolium castaneum (Herbst), to methyl and ethyl ester of C14-C18 fatty acid prepared from tallow were studied. Methyl octadecanoate and ethyl octadecanoate discouraged aggregation of beetle adults. Ethyl 9.12-octadecadienoate acted as a repellent. Methyl pentadecanoate and ethyl tetradecanoate induced copulation at the level of 5 × 10(2) and 2 × 10(3) µg, respectively. The maximum response was observed at the level of 5 × 10(2) µg of the same compounds, and these compounds appear to stimulate males only.

Insecticide fingerprinting technique for detection and location of organochlorine insecticide residues in foods.

Insecticide fingerprinting technique enables the detection and location of DDT and HCH residues in vegetables through the development of green and prussian blue colors respectively. Cut vegetables are pressed against o-tolidine impregnated paper and exposed to sunlight where colored spots appear instantly. The studies on 18 vegetable varieties revealed the pesticide residues and their distribution in different tissues. This direct method is sensitive (0.3/micrograms for HCH & 0.5/micrograms for DDT) and has special applications in quality control laboratories and food industry.