Structure-specific amyloid precipitation in biofluids.

The composition of soluble toxic protein aggregates formed in vivo is currently unknown in neurodegenerative diseases, due to their ultra-low concentration in human biofluids and their high degree of heterogeneity. Here we report a method to capture amyloid-containing aggregates in human biofluids in an unbiased way, a process we name amyloid precipitation. We use a structure-specific chemical dimer, a Y-shaped, bio-inspired small molecule with two capture groups, for amyloid precipitation to increase affinity. Our capture molecule for amyloid precipitation (CAP-1) consists of a derivative of Pittsburgh Compound B (dimer) to target the cross ß-sheets of amyloids and a biotin moiety for surface immobilization. By coupling CAP-1 to magnetic beads, we demonstrate that we can target the amyloid structure of all protein aggregates present in human cerebrospinal fluid, isolate them for analysis and then characterize them using single-molecule fluorescence imaging and mass spectrometry. Amyloid precipitation enables unbiased determination of the molecular composition and structural features of the in vivo aggregates formed in neurodegenerative diseases.

Heterogeneous precipitation mediated heterogeneous nanostructure enhances strength-ductility synergy in severely cryo-rolled and annealed CoCrFeNi2.1Nb0.2 high entropy alloy.

Possibilities of enhancing mechanical properties of brittle intermetallic containing high entropy alloys (HEAs) using novel processing and microstructural design strategies were investigated in the present work. For this purpose, homogenized CoCrFeNi2.1Nb0.2 HEA consisting of FCC matrix and complex Laves phase particles was successfully processed by severe cold- or cryo-rolling to 90% reduction in thickness followed by annealing (800 °C/1 hour(h)). As compared to cold-rolling, cryo-rolling resulted in a finer lamellar nanostructure and decidedly greater fragmentation of the Laves phase. Upon annealing, the cold-rolled HEA showed a recrystallized FCC matrix dispersed with D019 structured ε nano-precipitates. In contrast, the finer nanostructure and greater driving force for accelerated precipitation of profuse nano-precipitates at the early stages of annealing inhibited recrystallization in the cryo-rolled HEA and resulted in the formation of heterogeneous microstructure consisting of retained deformed and recrystallized regions. The novel heterogeneous microstructure of the cryo-rolled and annealed HEA resulted in a remarkable enhancement in strength-ductility synergy. The present results indicated that cryo-rolling could be used as an innovative processing strategy for tailoring heterogeneous microstructure and achieving novel mechanical properties.

Nanostructuring with Structural-Compositional Dual Heterogeneities Enhances Strength-Ductility Synergy in Eutectic High Entropy Alloy.

A lamellar (L12 + B2) AlCoCrFeNi2.1 eutectic high entropy alloy (EHEA) was severely deformed by a novel hybrid-rolling process. During hybrid-rolling, the deformation was carried out in two stages, namely cryo-rolling followed by warm-rolling at 600 °C. The strain (ε) imparted in each of these steps was identical ~1.2, resulting in a total strain of ε~2.4 (corresponding to 90% reduction in thickness). The novel processing strategy resulted in an extremely heterogeneous microstructure consisting of retained lamellar and transformed nanocrystalline regions. Each of these regions consisted of different phases having different crystal structures and chemical compositions. The novel structure-composition dual heterogeneous microstructure originated from the stored energy of the cryo-rolling which accelerated transformations during subsequent low temperature warm-rolling. The dual heterogeneous microstructure yielded an unprecedented combination of strength (~2000 MPa) and ductility (~8%). The present study for the first time demonstrated that dual structure-composition heterogeneities can be a novel microstructural design strategy for achieving outstanding strength-ductility combination in multiphase high entropy alloys.

Microstructural Characterization by Automated Crystal Orientation and Phase Mapping by Precession Electron Diffraction in TEM: Application to Hot Deformation of a γ-TiAl-based Alloy.

Microstructural evolution of a hot deformed γ-TiAl-based Ti-45Al-8Nb-2Cr-0.2B (at.%) alloy has been studied using an advanced characterization technique called automated crystal orientation and phase mapping by precession electron diffraction carried out in a transmission electron microscope (with a NanoMEGAS attachment). It has been observed that the technique, having a capability to recognize diffraction patterns with improved accuracy and reliability, is particularly suitable for characterization of complex microstructural features evolved during hot deformation of multiphase (α2 + γ + ß)-based TiAl alloys. Examples of coupled orientations and phase maps of the present alloy demonstrate that the accurate reproduction of the very fine lamellar structure (α2 + γ + γ) is feasible due to its inherent high-spatial resolution and absence of a pseudo-symmetry effect. It enables identification of salient features of γ-TiAl deformation behavior in terms of misorientation analyses (GAM, GOS, and KAM) and transformation characteristics of very fine lamellar constituent phases. Apart from conventional strain analyses from the orientation database, an attempt has been made to image the dislocation sub-structure of γ-phases, which supplements the deformation structure evaluation using this new technique.

Simultaneous Strength-Ductility Enhancement of a Nano-Lamellar AlCoCrFeNi2.1 Eutectic High Entropy Alloy by Cryo-Rolling and Annealing.

Nano-lamellar (L12 + B2) AlCoCrFeNi2.1 eutectic high entropy alloy (EHEA) was processed by cryo-rolling and annealing. The EHEA developed a novel hierarchical microstructure featured by fine lamellar regions consisting of FCC lamellae filled with ultrafine FCC grains (average size ~200-250 nm) and B2 lamellae, and coarse non-lamellar regions consisting of ultrafine FCC (average size ~200-250 nm), few coarse recrystallized FCC grains and rather coarse unrecrystallized B2 phase (~2.5 µm). This complex and hierarchical microstructure originated from differences in strain-partitioning amongst the constituent phases, affecting the driving force for recrystallization. The hierarchical microstructure of the cryo-rolled and annealed material resulted in simultaneous enhancement in strength (Yield Strength/YS: 1437 ± 26 MPa, Ultimate Tensile Strength/UTS: 1562 ± 33 MPa) and ductility (elongation to failure/ef ~ 14 ± 1%) as compared to the as-cast as well as cold-rolled and annealed materials. The present study for the first time demonstrated that cryo-deformation and annealing could be a novel microstructural design strategy for overcoming strength-ductility trade off in multiphase high entropy alloys.

Dark Matter Search Results from the PICO-60 C_{3}F_{8} Bubble Chamber.

New results are reported from the operation of the PICO-60 dark matter detector, a bubble chamber filled with 52 kg of C_{3}F_{8} located in the SNOLAB underground laboratory. As in previous PICO bubble chambers, PICO-60 C_{3}F_{8} exhibits excellent electron recoil and alpha decay rejection, and the observed multiple-scattering neutron rate indicates a single-scatter neutron background of less than one event per month. A blind analysis of an efficiency-corrected 1167-kg day exposure at a 3.3-keV thermodynamic threshold reveals no single-scattering nuclear recoil candidates, consistent with the predicted background. These results set the most stringent direct-detection constraint to date on the weakly interacting massive particle (WIMP)-proton spin-dependent cross section at 3.4×10^{-41} cm^{2} for a 30-GeV c^{-2} WIMP, more than 1 order of magnitude improvement from previous PICO results.

Non-migratory tumorigenic intrinsic cancer stem cells ensure breast cancer metastasis by generation of CXCR4(+) migrating cancer stem cells.

Although the role of metastatic cancer stem cells (mCSCs) in tumor progression has been well documented, our study reveals a hitherto unidentified role of tumorigenic intrinsic CSCs (iCSCs) in breast cancer metastasis. We show that unlike highly migratory mCSCs residing in the breast tumor disseminating/peripheral regions, iCSCs populate the inner mass of the tumor and are non-migratory. However iCSCs, via paracrine signaling, induce conversion of non-stem cancer cells to CSCs that (i) are identical to the previously reported mCSCs, and (ii) in contrast to iCSCs, express chemokine receptor, chemokine (C-X-C motif) receptor 4 (CXCR4), which is crucial for their metastatic potential. These mCSCs also demonstrate high in vivo tumorigenicity. Physical non-participation of iCSCs in metastasis is further validated in vivo, where only mCSCs are found to exist in the metastatic sites, lymph nodes and bone marrow, whereas the primary tumor retains both iCSCs and mCSCs. However, iCSCs ensure metastasis since their presence is crucial for deliverance of highly metastatic CXCR4(+) mCSCs to the migrating fraction of cells. Cumulatively, these results unveil a novel role of iCSCs in breast cancer metastasis as parental regulators of CXCR4(+) mCSCs, and highlight the therapeutic requisite of targeting iCSCs, but not CXCR4(+) mCSCs, to restrain breast cancer metastasis from the root by inhibiting the generation of mCSCs from iCSCs. Considering the pivotal role of iCSCs in tumor metastasis, the possibility of metastasis to be a 'stem cell phenomena' is suggested.

Epidemiology and antifungal susceptibility of Candida species in a tertiary care hospital, Kolkata, India.

BACKGROUND AND PURPOSE: The incidence of fungal infection as well as candidemia has increased significantly, contributing to morbidity and mortality in the developed countries. The alarming increase in infections with multidrug resistant bacteria is due to overuse of a broad spectrum antimicrobials, which leads to over growth of Candida spp.; thus, enhancing its opportunity to cause the disease. A shift has been observed in the relative frequency of each Candida spp. Antifungal agents available for the treatment of systemic and invasive candidiasis are restricted to polyenes, allylamines, azoles, and the recent echinocandin class of molecules. In the past few decades, the incidence of resistance to antifungal treatment of Candida spp. has increased rapidly, which is of serious concern for healthcare professionals. Studies on prevalence of infections and antifungal susceptibility testing can help with deciding on clinical strategies to manage this problem. Herein, we aimed to identify the epidemiology of Candida spp. among blood culture isolates and to investigate the susceptibility pattern of these species to antifungal agents. MATERIALS AND METHODS: Candida spp. were isolated from blood cultures from 70 patients in a tertiary care hospital, Kolkata, India. The growth of Candida spp. on sabouraud dextrose agar was confirmed by Gram staining, where gram-positive budding fungal cells were observed. The species identification as well as antifungal susceptibility testing were performed with VITEK 2 compact automated system using VITEK-2 cards for identification of yeast and yeast-like organisms) ID-YST card). Antifungal susceptibility testing was carried out with VITEK 2 fungal susceptibility card (AST-YS07 kit). RESULTS: Out of 70 samples, Candidaalbicans were isolated from 34 (%48.57) samples. The remaining 36 (%51.43) were non-albicans Candida ) NAC). Out of 34 C.Albicans, antifungal susceptiblity was detected in 28 isolates, all of which were sensitive to fluconazole (FLC .( Resistance to amphotericine B) AMP), flucytosine (5FC), voriconazole (VRC), and itraconazole (ITC) was observed in, %44.12 , %52.94 , %8.82 and %17.65 of the cases, respectively. For other Candida spp. (other than C.Albicans ,( antifungal susceptibility was evaluated for 36 isolates, among which resistance to AMP, FLC , 5FC, VRC , and ITC was found in, %30.56 , %61.11 , %33.33 , %19.44 and %38.89 cases, respectively. CONCLUSION: Species-level identification of Candida and their antifungal sensitivity testing should to be performed to achieve better clinical result and to select an appropriate and effective antifungal therapy. High resistance to antifungal agents is an alarming sign to the healthcare professionals.

Dark Matter Search Results from the PICO-2L C3F8 Bubble Chamber.

New data are reported from the operation of a 2 liter C3F8 bubble chamber in the SNOLAB underground laboratory, with a total exposure of 211.5 kg days at four different energy thresholds below 10 keV. These data show that C3F8 provides excellent electron-recoil and alpha rejection capabilities at very low thresholds. The chamber exhibits an electron-recoil sensitivity of <3.5×10(-10) and an alpha rejection factor of >98.2%. These data also include the first observation of a dependence of acoustic signal on alpha energy. Twelve single nuclear recoil event candidates were observed during the run. The candidate events exhibit timing characteristics that are not consistent with the hypothesis of a uniform time distribution, and no evidence for a dark matter signal is claimed. These data provide the most sensitive direct detection constraints on WIMP-proton spin-dependent scattering to date, with significant sensitivity at low WIMP masses for spin-independent WIMP-nucleon scattering.

Survey and census of hoolock gibbon (Hoolock hoolock) in the Inner-Line Reserve Forest and the adjoining areas of Cachar district, Assam, India.

A detailed survey of Hoolock hoolock was carried out in the Inner-Line Reserve Forest and the adjoining areas of Cachar district of southern Assam, India, from July 2010 to December 2011. About 150 km2 of the area was covered. In direct sighting, groups and individuals were counted in 7 localities (39.7 km2). Only 3.96 km2 of the actual forest area were occupied by these gibbons. Nine family groups and a solitary subadult, 33 individuals in all, made up the total count. Of these, adult males and females comprised 54.5% of the population while the subadults, juveniles and infants comprised 27.3, 12.1 and 6.1%, respectively. Each family group's home range was 0.31-0.51 km2. Of the 7 localities, only 1 had more than 1 family group. Habitat destruction and diverse threats to the hoolock gibbon in this area are examined in this paper.

The enigmatic Arunachal macaque: its biogeography, biology and taxonomy in Northeastern India.

The purpose of this study was to determine the taxonomic status of an unidentified enigmatic macaque seen by scientists since the late 1990s in Arunachal Pradesh, India. We surveyed 49 troops of enigmatic macaques in four districts of Arunachal Pradesh. The population studied is from the macaque sinica-group as defined by the reproductive organs. The main species-separating trait in the sinica-group is tail length to head and body length ratio that decreases with latitude and elevation. We gathered data on morphology, pelage descriptions, tail to head and body ratios and tail to hind foot ratios from photos and live animals (43 individuals from 36 areas) within the range of and between the two subspecies of the Assamese macaque (Macaca assamensis). We compared the data to six western Assamese macaques and studies of Assamese macaques and related species. We found great variability in tail length, pelage color, facial skin color, and facial and hair patterns. The tail/head-body and tail/foot ratios, although varied, supported the hypothesis that these enigmatic forms were part of a population of Assamese macaques found in the gap between the two subspecies ranges and were not a new species as described earlier. Instead, we found evidence that darker pelage, larger body size, and shorter tails occur at higher elevations and latitudes similar to the general trend in the sinica-group's adaptations to colder climates. Thus, the population may be important for its variation, throwing light on the speciation process and how the northern species of Tibetan macaques evolved from an ancestor similar to the Assamese macaques as adaptations to a colder climate.

Sweat gland carcinoma - a case report.

A 54 years male presented with painless, indolent ulcers in the left axilla in an area approximately of 5 cm diameter. They were fixed to the underlying structures. Some lymph nodes eg, pectoral, apical and central groups were also enlarged. Incision biopsy for the margin of the ulcer was done. It showed adenocarcinoma of the sweat glands. Wide local incision with 2 cm margin of surrounding skin was performed. The patient was discharged after a course of adjuvant radiotherapy and is doing well.

Haematological presentation of systemic lupus erythematosus.

Haematological manifestations are quite common in systemic lupus erythematosus (SLE) but bone marrow aplasia and secondary myelofibrosis are rare manifestations. We report a case of 45 years old male patient who presented with fever, malaena and anaemia without any clinical features of SLE. He had patches of vitiligo for 25 years for which he was applying psoralen with sunlight exposure. This probably precipitated SLE in this patient. The patient presented with pancytopenia which was due to a combination of Comb's positive haemolytic anaemia and myelofibrosis.

Sarcomatoid squamous cell carcinoma: a long-standing case.

Sarcomatoid, or spindle cell squamous cell carcinoma (SCSC), is relatively uncommon, but may be encountered. It poses a challenge in differential diagnosis that includes other spindle cell neoplasms. We present a case where the lesion existed for 20 years and raised the potential of chronicity as a factor in inducing spindle cell morphology. Detailed immunohistochemical features are demonstrated, and discussion of the differential diagnosis is offered. The patient was an 89-year-old African-American female with an exophytic and polypoid mass of the right upper arm measuring 5.5 x 5.5 x 3.0 cm. The mass had been present for the last 20 years, and was gradually and very slowly increasing in size. After refusing surgery several times, she finally agreed to have an excision. The tumor proved to be SCSC.

Concepts and strategies for scaling up focused prevention for sex workers in India.

OBJECTIVES: To describe the concepts, strategies and field results of a project to scale up prevention programmes and services for female sex workers (FSWs) in Karnataka, India. METHODS: A strategy was developed to scale up urban sex worker interventions in 18 districts in the southern Indian state of Karnataka. Macro-level coverage objectives were defined by mapping the urban locations where FSWs operate and estimating their population size. Prevention programmes were initiated in the urban locations that contained at least 90% of the estimated urban FSW population in each district. Within each location, a micro-planning process was used by FSW peer educators and outreach workers to design local outreach and service delivery plans. RESULTS: An estimated 48 973 FSWs were distributed across 1551 locations and 6232 spots. Outreach was conducted by 1043 peer educators. Services were provided through 170 drop-in centres, 93 programme-run clinics, 110 outreach clinics and 157 referral clinics. Within the first 3 years of the programme the cumulative number of individual FSWs contacted at least once was >78 000, with monthly contact established with 81% of the in situ population; >45 000 FSWs had visited a clinic and >10 000 visited monthly. Direct and indirect condom distribution by the programme amounted to more than 30 per contacted FSW, which is estimated to meet the condom requirement. CONCLUSIONS: A strategy that involves geographically defined coverage and micro-level outreach planning can rapidly and effectively provide outreach and services to large dispersed FSW populations.

Follicular carcinoma of thyroid with synchronous bony and soft tissue metastases.

A 65 years old lady presented with a large thyroid swelling along with a soft, non-tender mass over the left parietal region and a small, hard, non-tender swelling over the volar aspect of right forearm. FNAC reports from the thyroid gland and the swellings over head and forearm all revealed features of follicular neoplasm. Excision biopsy of the forearm swelling showed histology of metastatic poorly differentiated follicular adenocarcinoma of thyroid. The patient underwent total thyroidectomy followed by excision of the mass over the head, a week later. She was discharged in favourable condition with advice to attend oncology department for subsequent management. She is yet to attend surgical outdoor for follow-up.

Prospective evaluation of modified Alvarado score for diagnosis of acute appendicitis.

Modified Alvarado scoring system was evaluated regarding its usefulness in the early diagnosis of acute appendicitis and in reduction of the incidence of negative appendicectomies. One hundred and ten patients with a provisional diagnosis of acute appendicitis were studied prospectively at R G Kar Medical College and Hospital, Calcutta over a period of 10 months. High score was found to be a dependable aid both in the pre-operative diagnosis of acute appendicitis and in the reduction of negative appendicectomies in men and children but the same was not true for women who had a high false positive rate for acute appendicitis.

Bovine herpesvirus 5 (BHV-5) Us9 is essential for BHV-5 neuropathogenesis.

Bovine herpesvirus 5 (BHV-5) is a neurovirulent alphaherpesvirus that causes fatal encephalitis in calves. In a rabbit model, the virus invades the central nervous system (CNS) anterogradely from the olfactory mucosa following intranasal infection. In addition to glycoproteins E and I (gE and gI, respectively), Us9 and its homologue in alphaherpesviruses are necessary for the viral anterograde spread from the presynaptic to postsynaptic neurons. The BHV-5 Us9 gene sequence was determined, and the predicted amino acid sequence of BHV-5 Us9 was compared with the corresponding Us9 sequences of BHV-1.1. Alignment results showed that they share 77% identity and 83% similarity. BHV-5 Us9 peptide-specific antibody recognized a doublet of 17- and 19-kDa protein bands in BHV-5-infected cell lysates and in purified virions. To determine the role of the BHV-5 Us9 gene in BHV-5 neuropathogenesis, a BHV-5 Us9 deletion recombinant was generated and its neurovirulence and neuroinvasive properties were compared with those of a Us9 rescue mutant of BHV-5 in a rabbit model. Following intranasal infection, the Us9 rescue mutant of BHV-5 displayed a wild-type level of neurovirulence and neural spread in the olfactory pathway, but the Us9 deletion mutant of BHV-5 was virtually avirulent and failed to invade the CNS. In the olfactory mucosa containing the olfactory receptor neurons, the Us9 deletion mutant virus replicated with an efficiency similar to that of the Us9 rescue mutant of BHV-5. However, the Us9 deletion mutant virus was not transported to the bulb. Confocal microscopy of the olfactory epithelium detected similar amounts of virus-specific antigens in the cell bodies of olfactory receptor neuron for both the viruses, but only the Us9 rescue mutant viral proteins were detected in the processes of the olfactory receptor neurons. When injected directly into the bulb, both viruses were equally neurovirulent, and they were transported retrogradely to areas connected to the bulb. Taken together, these results indicate that Us9 is essential for the anterograde spread of the virus from the olfactory mucosa to the bulb.

A comparative study on the physical characteristics and cooking quality parameters of commercial brands of Basmati rice.

Randomly selected samples of three popular commercial brands of Basmati rice (designated as A, B and C) available in Mumbai city were studied with respect to their proximate composition and differences in their quality evaluated in terms of length by breadth and elongation ratio, differential length by breadth ratio, water uptake, loss of solids in cooking water, swelling index and volume expansion ratio. The l/b ratio and elongation ratio varied between 4.47 to 4.81 and 1.45 to 1.59 respectively. The differential length by breadth ratio showed a wide variation from 5.88 to 9.48 among the three brands studied. Brand A recorded the highest length by breadth and elongation ratio whereas brand C recorded the highest differential length by breadth ratio. The water uptake values and the loss of solids in the cooking gruel varied between 172 to 262 and 0.87 to 2.70 (g/100 g of rice) respectively, among the three commercial brands. The swelling index and the volume expansion ratio did not show much variation among the three brands ranging between 28 to 31 for the former and 2.00 to 2.21 for the latter. Based on the differential length by breadth ratio, water uptake and swelling index values, which are the most reliable quality indices; brand C was found to be the best followed by brands A and B. The study of the texture of the cooked rice grains indicated that brand C had the softest grains, which was in conformity with the findings in cooking quality characteristics.