Search

Aberrant CpG-island methylation affects ovarian cancer progression. The promotor methylation changes at tumor suppressive genes in ovarian cancer stromal progenitor cells (OCSPCs) and epithelial ovarian cancer (EOC) tissues and their clinical implication remains unexplored. We systemically analyzed the promoter methylation status of 40 tumor suppressor genes (TSGs) associated with cancer in paired epithelial-like and mesenchymal-like OCSPCs and ovarian cancer cells by methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA). The effect of DNA methylation on gene expression was confirmed using qRT-PCR. The differential frequencies of TSGs' promoter methylation among matched epithelial-like or mesenchymal-like OCSPCs from tissues and ascites and ovarian cancer tissues were further validated in cancer tissues and correlated with clinicopathological features and survival outcomes of patients. According to the promoter methylation frequencies of the 40 TSGs, promoters of RASSF1A were the only significantly hypomethylated in epithelial-like OCSPCs from tissues than those from ascites and bulk tumor cells (0% vs 38% vs 45%, P=0.039 by Fisher's exact test). The most frequencies at promotor hypermethylation of TSGs in mesenchymal-like OCSPCs from ascites which processed aggressiveness were CDKN2B (73%) followed by CCND2 (45%) and RASSF1A (45%). Forty-three percent (47/110) of RASSF1A and 45% of CCND2 were validated as a frequently hypermethylated gene in an independent set of 110 EOC tissues in contrast to none (0/60) and 12% (10/60) of benign ovarian cysts (both P0.001). Functional experiments revealed overexpression of CCND2 or CDKN2B in MSc-OCSPCs decreases EMT, invasion, and spheroid formation in EOC, and abolishes DNMT1 and COL6A3 expression. However, for the expected 5-year overall survival (OS) for patients with methylated RASSF1A, CCND2, and CDKN2B, only RASSF1A was significantly worse than those without methylated RASSF1A (56% vs 80%, p=0.022). Taken together, overexpression of CCND2 and CDKN2B decreased the aggressiveness of mesenchymal-like OCSPCs from ascites which may represent a potential therapeutic target for EOC. Promotor hypomethylation at RASSF1A in OCSPCs from EOC tissues and changes to hypermethylation of EOC and OCSPCs from ascites could predict poor survival outcomes for EOC patients compared to without those changes of CCND2 and CDKN2B.

Syndiotactic poly(4-methyl-1-pentene) (sP4M1P)-based stereoregular diblock copolymers, namely sP4M1P-b-polystyrene and sP4M1P-b-polymethylmethacrylate, were prepared from an α-bromoester-capped sP4M1P macroinitiator, which was chain extended with styrene and methyl methacrylate, respectively, via the atom transfer radical polymerization reaction. The α-bromoester-capped sP4M1P was generated by the esterification of hydroxyl-capped sP4M1P with α-bromoisobutyryl bromide. The hydroxyl-capped sP4M1P was synthesized by inducing a selective chain transfer reaction to aluminum during the syndiospecific polymerization of 4-methyl-1-pentene in the presence of a syndiospecific metallocene catalyst. As stereoregular diblock copolymers are difficult to prepare using existing methods, the current study offers an effective process for the preparation of sP4M1P-based stereoregular diblock copolymers. These copolymers were found to have well-defined architectures and they can undergo molecular self-assembly into ordered nanostructures, as evidenced by small-angle X-ray scattering analyses.

Ovarian clear cell cancer stem-like/spheroid cells (OCCCSCs) were associated with recurrence, metastasis, and chemoresistance in ovarian clear cell carcinoma (OCCC). We evaluated the anti-tumor effects of 5-aza-2-deoxycytidine (5-aza-dC) combined with everolimus (RAD001) on human OCCC. We investigated parental OCCCSCs and paclitaxel-resistant cell lines derived from OCCCSCs in vitro and in vivo. A Western blot analysis showed that the 5-aza-dC and RAD001 combination therapy was associated with the COL6A3-AKT-mTOR pathway. The OCCCSCs expressed high levels of stemness markers: CD117, ALDH1, NANOG, OCT4, and CD133. The 5-aza-dC and RAD001 combination inhibited proliferation and survival with up to 100-fold more potency in OCCCSCs compared to OCCC cells. This combination showed significant anti-tumor activity; it preferentially diminished OCCCSC stemness levels and spheroid numbers in vitro. Limiting dilution assays showed that OCCCSCs possessed tumor-initiating capacity. The 5-aza-dC and RAD001 combination significantly enhanced the inhibition of tumor growth compared to the 5-aza-dC or RAD001 alone. OCCCSCs showed higher expression levels of COL6A3, phospho-AKT, phospho-mTOR, and phospho-Rictor compared to OCCCs. Silencing COL6A3 or abolishing the phospho-AKT-mTOR-Rictor pathway with 5-aza-dC and RAD001 treatment further enhanced OCCCSC apoptosis and reduced OCCCSC stemness. In conclusion, 5-aza-dC combined with RAD001 effectively controlled OCCC and OCCCSC growth by inhibiting the COL6A3-AKT-mTOR pathway.

The authors would like to make a correction to their published paper [...]

The impact of the new International Association for the Study of Lung Cancer pathology committee grading system for advanced lung adenocarcinoma (LADC) on survival is unclear, especially in Asian populations. In this study, we reviewed the prognostic outcomes of patients with late-stage disease according to the new grading system. We reviewed 136 LADC cases who underwent a small biopsy from 2007 to 2018. Tumors were classified according to the new grading system for LADC. Baseline characteristics (age, sex, smoking status, body mass index, and driver gene mutations) were analyzed. Kaplan&ndash, Meier and Cox regression analyses were used to determine correlations with the new grading system and prognosis. Patients with poorly differentiated adenocarcinoma were significantly correlated with a poor progression-free survival (PFS) (p = 0.013) but not overall survival (OS) (p = 0.154). Subgroup analysis showed that wild-type EGFR patients with poorly differentiated adenocarcinoma treated with chemotherapy had significantly worse PFS (p = 0.011). There was no significant difference in survival among the patients with epidermal growth factor receptor mutations who were treated with tyrosine kinase inhibitors. Patients aged &gt, 70 years and those with a BMI &le, 25 kg/m2 and wild-type patients had significantly worse OS in both univariate (HR = 1.822, p = 0.006, HR = 2.250, p = 0.004, HR = 1.537, p = 0.046, respectively) and multivariate analyses (HR = 1.984, p = 0.002, HR = 2.383, p = 0.002, HR = 1.632, p = 0.028, respectively). Despite therapy, patients with poorly differentiated tumors still fared worse than those with better differentiated tumors. No differences were found among the EGFR mutations treated with TKI. Our findings highlight that the therapeutic regimen should be adjusted for EGFR Wild-type patients with poorly differentiated adenocarcinoma treated with chemotherapy to provide better outcomes.

Bananas are one of the most popular fruits worldwide, producing over 100 million tons in 2013. Banana peel, a byproduct that represents 40% of the entire banana’s weight, is rich in bioactive compounds and has a high antioxidant capacity. Therefore, the aim of this study is to use banana peel powder (BPP) as an antioxidant broiler feed additive. The bioactive compound and antioxidant capacity of BPP were determined. The results showed that BPP contained 872.7 GAE/100 g DM of total phenolic content, and gallocatechin was detected at a concentration of 91 mg/100 g DM, and BPP’s ferrous chelating ability, ferric ion reducing ability power, Trolox equivalent antioxidant capacity, and 1,1-diphenyl-2-picrylhydrazyl scavenging capacity indicated its high antioxidant capacity. A total of 216 one-day-old Ross 308 female broiler chicks were randomly assigned to one of four groups that received the basal diet supplemented with 0% (control), 0.5%, 1%, or 2% dry BPP for 35 d. The results showed that the 0.5% BPP-supplemented group had increased serum SOD activity and lowered serum malondialdehyde levels, while the liver mRNA expression of Nrf2, SOD, CAT, GPx, and HO-1 were upregulated in the 0.5% and 1% BPP-supplemented groups. In conclusion, the current study shows that BPP has a potent antioxidant capacity and dietary supplementation with 0.5% BPP enhances antioxidative status, suggesting that BPP has potential as an antioxidant feed additive.Highlights Banana peel has abundant bioactive compounds and antioxidant capacity. Banana peel enhance the antioxidant status of broilers by up-regulating Nrf2/ARE associated genes. Banana peel has potential as an antioxidant feed additive in broiler chickens.

In this study we prepared a novel triblock copolymer (tBCP) through ring-opening polymerization (ROP) with e-caprolactone (e-CL) and two specific atom transfer radical polymerizations (ATRPs), with n-butyl acrylate (nBA) and methyl methacrylate (MMA), respectively (herein, a supplemental activator and reducing agent and halogen-exchange ATRPs were employed). These chain-extending polymerizations were chosen to achieve controlled/living polymerization. This strategy afforded a well-defined ABC-type PCL-b-PnBA-b-PMMA tBCP (Mn, GPC = 56 600; PDI = 1.18) comprising two external miscible blocks with an epoxy monomer and a middle soft block. After curing with the diglycidyl ether of bisphenol-A and 4,4′-methylenedianiline, the miscibility of the resulting epoxy thermoset (ET)/tBCP composites was examined using Fourier transform infrared spectroscopy, differential scanning calorimetry, and thermogravimetric analysis. On further examining the ET/tBCP microstructures using transmission electron microscopy and small-angle X-ray scattering, we observed transitions with a long range order—from lamellar to core/shell cylinder to short core/shell cylinder morphologies—upon increasing the amount of ET. The domain sizes obtained after the curing reaction were in the range of approximately 30–60 nm.

Stereoregular side-chain photoluminescent (PL) polymers were synthesized via the stereospecific polymerization of 9,9-dibutyl-2-(4-vinylphenyl)-9H-fluorene. The resulting isotactic, syndiotactic and atactic poly(9,9-dibutyl-2-(4-vinylphenyl)-9H-fluorene) polymers were used as stereoregular polymer samples for examining the configurational tacticity effect on their PL properties. Our results revealed a unique tacticity-dependent effect as observed by an obvious red-shifting of the PL emission by increasing the degree of isotacticity of these stereoregular polymers; despite the ultraviolet spectra of these polymers exhibited similar absorption patterns. This study provides a new structural design for the syntheses of PL polymers.

Medulloblastoma (MB) is the most common pediatric malignant brain tumor and patients with high-risk or recurrent MB respond poorly to current therapies, and have a higher related mortality. For this reason, potential molecules related to MB need be identified in order to develop targets for the development of novel therapeutics. In the present study, we compared MB microarray data obtained using different microarray systems and significant targets were selected by gene annotation and enrichment analysis. Genes for soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) annotated with the function 'vesicle' were identified and one of these proteins, synaptosomal-associated protein 25 (SNAP25), was found to have significantly lower expression levels in MB. In addition, SNAP25 was detected in a very low number of MB cells as shown by western blot analysis and immunohistochemical analyses of archived and formalin-fixed/paraffin-embedded human MB specimens. We found that SNAP25 altered the morphology and the chemotherapeutic effects of arabinofuranosyl cytidine (Ara-C) on SNAP25-expressing MB cells. On the whole, our data indicate that the expression of SNAP25 is crucial for dendrite formation and is associated with the effects of targeted chemotherapy. The detection of SNAP25 expression in MB cells may thus be essential for the chemotherapeutic application of Ara-C.

Single, double, and coincident crystallizations under hard or soft confinement are all carried out using a single type of syndiotactic poly(p-methylstyrene)-block-poly(l-lactide) (sPPMS–PLLA) block copolymers. The single crystallization of sPPMS matrix can lead to the disordered arrangement of hexagonally packed PLLA cylinders under soft confinement. In contrast, the lamellar nanostructure remained unchanged regardless of the PLLA crystallization under hard or soft confinement. Crystallization-induced morphological transitions from the confined monosized lamella to the metastable dual-sized lamella and finally to the breakout morphology are evident by transmission electron microscopy and small-angle X-ray scattering. The dual-sized lamella is attributed to the thermodynamically and kinetically controlled nanocrystallite growth templating along the ordered microphase separation. Despite crystalline sequences, the double-crystallized morphologies are determined by the first-crystallized event even though th...

KRAS and BRAF mutations are frequently detected in cases of colorectal cancer (CRC). The microsatellite status of patients with CRC and mutated KRAS/BRAF is important when determining cancer therapy. In the present study, the microsatellite status and genetic polymorphisms of KRAS (codons 12 and 13) and BRAF (V600E) were characterized in CRC tissue. The mismatch repair activity and oncogenic potential of KRAS were assessed by immunoblots from two KRAS-mutated CRC cell lines, SW480 and HCT116, with different microsatellite statuses, following treatment with 5-fluorouracil (5-FU) and oxaliplatin. Of all the 205 patients with CRC enrolled in the present study, 31.2% (64 of 205) had a KRAS or BRAF mutation, and 79.7% (51 of 64) of these patients with a KRAS/BRAF mutation exhibited microsatellite stability (MSS), indicating that microsatellite status is correlated with KRAS/BRAF mutation (P=0.027). A higher proportion (39.0%, 41 of 105) of elderly patients (≥62.6 years) had mutated KRAS or BRAF than younger patients (

This study presents an optical module which collects wide-angle solar power to enhance the power generation efficiency of Photovoltaic (PV) panels. The ultra-wide angle optical module is able to replace the traditional mechanical solar tracking system. This module is composed of an array of light guiding units, and each unit includes two parts which are asymmetrical compound parabolic curve (ACPC) and the free-form curve collimator. The ACPC and collimator are combined to detect the power generation.

Background : Barrett’s esophagus (BE) is defined as a suspect endoscopic finding followed by pathologically proven specific intestinal metaplasia as opposed to the normal squamous epithelium of the esophagus. A new BE classification, the Prague C and M criteria, was proposed in 2006 to define the extent of BE based on the circumference (C) and maximum diameter (M) of the lesion. Original Research Article

In this study we blended 4-(4-hexadecylphenylazo)pyridine (AzoPy-C16), synthesized through facile diazonium and monoetherification reactions, with polytyrosine (PTyr) to form comb-like polypeptide supramolecular complexes stabilized through hydrogen bonding between the pyridyl ring of each AzoPy-C16 unit and the OH groups of PTyr. The secondary structure of PTyr transformed from an α-helical to a β-sheet conformation upon the addition of AzoPy-C16, because the long alkyl chains of the AzoPy-C16 units disrupted the weak intramolecular hydrogen bonds between the CO and NH groups in the α-helical conformation, as determined using Fourier transform infrared (FTIR) and circular dichroism (CD) spectroscopy. Small-angle X-ray scattering (SAXS) and wide-angle X-ray diffraction (WAXD) suggested a hierarchical lamellae-within-lamellae structure for the PTyr/AzoPy-C16 supramolecular complex, featuring long-range-ordered lamellae arising from the antiparallel β-pleated sheet conformation of PTyr (d = 1.15 nm) within long-range-ordered lamellae arising from the alkyl groups of AzoPy-C16 units (d = 5.94 nm), oriented in a perpendicular manner. The d-spacing and long-range order of the lamellar structure formed from the alkyl groups decreased upon UV irradiation as the rod-like trans isomers of the AzoPy-C16 units transformed into V-shaped cis counterparts. This phenomenon also led to a change in the water contact angle of the supramolecular material, with the hydrophobic surface of PTyr/trans-AzoPy-C16 (94.2°) transforming to a hydrophilic surface for PTyr/cis-AzoPy-C16 (61.3°).

Manipulation of crystalline textures of biocompatible block copolymers is critical for the applications in the medical field. Here, we present the control of multiple-crystalline morphologies with flat-on chain orientation in biocompatible poly(ethylene oxide)-block-poly(e-caprolactone)-block-poly(l-lactide) (PEO–PCL–PLLA) triblock copolymer thin films using melt and solvent-induced crystallizations. Only single-crystalline morphologies of the first-crystallized blocks can be obtained in the melt-crystallized thin films due to the confinement effect. With solvent annealing by PCL-selective toluene, single-crystalline PLLA to double-crystalline PLLA/PCL and to triple-crystalline PLLA/PCL/PEO layered crystals in sequence are observed for the first time. With the control of solvent selectivity, different sequential crystallization involving first-crystallized PCL transferring to double-crystalline PCL/PLLA is obtained using PEO-selective n-hexanol for annealing. Surprisingly, the crystalline growth of the tr...

Background Lymphoepithelioma-like carcinoma (LELC) is a rare entity among thyroid tumors. Based on the limited number of case reports that exist, the association of Epstein–Barr virus (EBV) with primary thyroid LELCs seems inconsistent. Case presentation We present a confusing cytological case of lymphoepithelioma-like thyroid carcinoma with expression of EBV. The patient presented with a central neck mass and bilateral lymphadenopathy. Fine-needle aspiration cytology revealed three-dimensional and syncytial fragments of epithelioid cells accompanied by small lymphocytes. The surgical specimen of resected thyroid tumor disclosed typical histopathological features of LELC. Metastatic papillary carcinoma was also discovered in the metastatic lymph nodes. In situ hybridization for EBV-encoded RNA (EBER-ISH) was positive in the tumor cells. Negative immunoreactivity for TTF-1, Pax-8, and CD5 was observed. The patient is currently undergoing regular follow-up and is 1 year and 10 months postresection with no evidence of recurrence. Conclusions Long-term survival is discussed in relation to this variant of thyroid carcinoma, which might differ in behavior from anaplastic carcinoma. Further investigation is required to elucidate the clinical significance of EBV expression and progression of this unique variant of thyroid carcinoma.

Organic luminogens with aggregation-induced emission (AIE) properties have unusual emission behaviour in that their non- or weakly-emissive solutions can be tuned to emit strongly in the solution aggregated and solid states. As restricted molecular rotation of luminogens is the main mechanism leading to AIE activity, hydrogen bond (H bond) interactions are therefore used in organic and polymeric luminogens to impose efficient rotational restriction and to intensify the emission intensity of the AIE-active luminogens. Luminogenic polymers containing H-bonding groups and luminogenic blends consisting of H-bond donors and acceptors are therefore described in this review to evaluate the relationship between H bond interactions, rotational restriction and AIE-related emission intensity.

Our previous study showed that 5-aza-2-deoxycytidine (5-aza-dC) could inhibit tumor growth by enhancing the susceptibility of ovarian clear cell carcinoma (OCCC) to paclitaxel through decreasing AKT/mTOR expressions. The objective of the study is to evaluate the antitumor efficacy of everolimus (RAD001) and 5-aza-2-deoxycytidine (5-aza-dC) by targeting AKT/mTOR and EZH2 in OCCC. Paclitaxel-sensitive and resistant OCCC cell lines were established. In vitro proliferative and apoptotic assays and flow cytometry were performed. The expressions of EZH2, PIK3IP1, phospho-AKT, phospho-mTOR and phospho-Rictor in the OCCC cell lines were evaluated by Western blotting. In vivo animal experiments with RAD001 and 5-aza-dC were performed. RAD001 alone showed significant in vitro antitumor activity and inhibited in vivo tumor growth in paclitaxel-sensitive and resistant OCCC cells. In addition, 5-aza-dC enhanced the antitumor effects when combined with paclitaxel or RAD001 in both paclitaxel-sensitive and resistant tumors. Activation of phospho-AKT ser473 and PIK3IP1 was observed in RAD001-treated paclitaxel-sensitive and resistant OCCC cells. In contrast, inhibition of phospho-AKT ser473 and EZH2 was observed with RAD001 following 5-aza-dC treatment of paclitaxel-sensitive and resistant OCCC cells. Furthermore, RAD001 following 5-aza-dC enhanced apoptosis of paclitaxel-sensitive and resistant OCCC cells. RAD001 following 5-aza-dC may be a promising treatment strategy for the treatment of both chemo-sensitive and resistant OCCC. Further clinical studies are warranted.

This study presents an ultra-wide angle module on the fixed-type Photovoltaic (PV) panel to enhance the efficiency without a mechanical solar tracker. This ultra-wide angle module was composed of an array of light guiding unit, and each unit consisted of an asymmetrical compound parabolic curve (ACPC) and a free-form curve collimator. The ACPC was used to collect the misalignment and diffused light and the light was guided to the free-form curve collimator. The collimator provided the light for a smaller angle and better uniformity, and each light guiding unit was put in array for easily attaching to the PV panel. The simulation results show that this ultra-wide angle module had the optical efficiency above 80% when the misalignment angle was smaller than 20°. In addition, the commercial products are used for the experiment, and the results show that the CPC had 24% higher and the light guiding unit had 37% higher power generation than the naked PV panel.

Background Low-grade fibromyxoid sarcoma (LGFMS) is a rare soft tissue tumor that has a tendency to grow in the deep soft tissue of the trunk and extremities. Despite its benign appearance, the tumor has a high recurrence rate and metastatic potential. LGFMS in the perineal space is rare, and only a few cases have been reported. We present the first case of LGFMS to be located at the external anal sphincter. Case presentation A 27-year-old male patient admitted to our Surgical Department with perianal pain and swollen for a year. The digital rectal examination revealed a perianal mass. Oral metronidazole and analgesia were prescribed on suspicion of perianal abscess failed to alleviate the symptom; hence, the patient was scheduled for surgery. Intraoperative diagnosis revealed an encapsulated tumor in the external anal sphincter that extended from the perianal region orally to the pararectal space. The results of immunohistochemistry (MUC4 staining) and FUS gene rearrangement by fluorescence in situ hybridization confirmed the diagnosis of LGFMS. Conclusions This case is unique in terms of the location of the rare soft tissue tumor. Although LGFMS is considered low grade, its unpredictable behavior necessitates a long-term follow-up.

Summary Gastritis cystica polyposa is relatively rare and characterized by polypoid hyperplasia and cystic dilatation of the gastric glands in stomach. Most cases are related to previous gastric surgeries, but a few cases have been reported in unoperated stomachs. We present a 34-year-old man who had anemic symptoms with melena and exertional dyspnea for 3 weeks. He denied any surgical history. An esophagogastroduodenoscopy revealed diffuse giant folds extending from the lower to the upper body of the stomach with nodularity and no obvious bleeding site. A pathologic diagnosis of a punch biopsy specimen from the giant folds revealed only moderately active chronic inflammation with a high Helicobacter pylori density. After serial studies, the patient received a whole layer gastric biopsy during a laparoscopy. Gastritis cystica polyposa was diagnosed on the pathology report. Our present case highlights the rare clinical and endoscopic condition of gastritis cystica polyposa in an unoperated stomach.

According to size and intensities of leukocytes in third-harmonic-generation microscopy, we differentiate and count neutrophils, monocytes, and lymphocytes both ex vivo and in vivo. Results agreed well with those of conventional blood tests.

Based on third-harmonic-generation (THG) microscopy and a k-means clustering algorithm, we developed a label-free imaging cytometry method to differentiate and determine the types of human leukocytes. According to the size and average intensity of cells in THG images, in a two-dimensional scatter plot, the neutrophils, monocytes, and lymphocytes in peripheral blood samples from healthy volunteers were clustered into three differentiable groups. Using these features in THG images, we could count the number of each of the three leukocyte types both in vitro and in vivo. The THG imaging-based counting results agreed well with conventional blood count results. In the future, we believe that the combination of this THG microscopy-based imaging cytometry approach with advanced texture analysis of sub-cellular features can differentiate and count more types of blood cells with smaller quantities of blood.

Background Saussurea involucrata (Kar. et Kir.) commonly known as ‘snow lotus’ or ‘Xue Lian’ is an important plant in the traditional Chinese system of medicine. The plant contains flavonoids such as syringin and rutin. These compounds have been reported to be anti-rheumatic, anti-inflammatory and dilate blood vessels, lower blood pressure, prevent cardiovascular diseases, enhance immunity, and act as anti-aging, anti-cancer, and anti-fatigue agents. The species has become endangered due to the excessive collection of S. involucrata plants in the wild, slower plant growth and ecological destruction of natural habitats. There is a severe shortage of plant material, while the market demand is ever increasing. Hence, it is very important to apply tissue culture technique for plant propagation and production of the bioactive compounds of this species. Results Multiple shoot induction and proliferation in shoot base explants derived from in vitro raised seedlings of S. involucrata was achieved on 3/4 strength of Murashige and Skoog’s (MS) basal medium (MSBM) supplemented with 1.0 mg/L−1 BA and 1.5 mg/L−1 NAA. Rooting was induced in 100 % shoots cultured on 1/2X MSBM supplemented with 1.0 mg/L−1 IBA for one week and then transfer to auxin free medium. The plantlets could be acclimatized successfully by sachet technique and established in the greenhouse. Maximum callus induction and proliferation in leaf segments was achieved on 1/2X MSBM supplemented with 0.5 mg/L−1 BA, 0.5 mg/L−1 NAA, 0.4 % gelrite and on incubation at 20 °C. Container closures had an influence on the quality and quantity of callus and production of the active compounds. The HPLC analysis showed much higher syringin content in in vitro shoots and callus as compared to commercially available market crude drug. Conclusion The present study describes an in vitro culture protocol of Saussurea involucrata. The bioactive compounds, syringin and rutin could be produced through tissue culture technique without sacrificing the endangered Saussurea involucrata plants in the wild.

Additional file 5: Table S4. Frequency of methylation among stromal progenitor cells from ascites, cancerous tissues, and bulk tumor cells.

This chapter describes the different methodologies used for in vitro micropropagation and cell suspension culture in Gentiana sp. Low concentrations of auxins and kinetin enhanced the growth of suspension cells. Maximum growth in suspension cultures of Gentiana sp. was obtained at shaker speed of 80–100 rpm, pH 4.2–5.2, and a light intensity of 2.33 μE m−2 s−1. Attention is also given to the metabolite content of different plant organs of Gentiana sp. The aerial and underground parts of G. davidii contain the greatest amounts of gentiopicroside and swertiamarin. These findings may be implemented for micropropagation, germplasm conservation, and commercial cultivation, and their active principles analysis in other members of the Gentianaceae.

Giardiasis is an important waterand food-borne infectious disease that causes sporadic diarrhea in travelers and daycare center attendees worldwide. We report a 33-year-old diabetic mother with giardiasis presenting as malnutrition and severe weight loss from 85 kg to 40 kg over 6 months, who had close contact with her mentally impaired son at a sanatorium. The duodenal biopsy confirmed the Giardia trophozoites among the villi and crypts (Fig. 1). The architecture of the duodenal mucosa was well preserved. Electron microscopy confirmed the presence of Giardia trophozoites in the duodenal villi. She responded to metronidazole therapy, as her symptoms improved and she regained weight to 50 kg in 24 weeks. As few as 10e25 cysts can cause Giardia infection following the ingestion of cysts from contaminated water or food. Giardia cysts transform into trophozoites in the proximal small bowel, where trophozoites replicate on the mucosal surface and usually do not invade the mucosa. Trophozoites may directly invade or release toxic substances to damage the intestinal epithelium. Trophozoites transform into infectious cysts in the distal small intestine and colon.