Understanding the dynamics of Meloidogyne incognita infestation in pomegranate orchards of Himachal Pradesh, India (year 2018, 2019 and 2021) and its management strategies.

This study investigates the prevalence and dynamics of pomegranate wilt disease induced by Meloidogyne incognita across the Kullu, Mandi and Solan districts of Himachal Pradesh (India), revealed notable spatial and temporal variations in nematode populations and galling severity across the regions. The highest average nematode infestation of 9.25 % was observed at Nauni with highest counts of average second-stage juvenile (J2) larvae (449 larvae per 100 cc of soil) followed by Hurla (Kullu) with 7.42 % infestation. Correlation analysis reveals a strong positive relationship between larval population and galling severity suggesting a potential link between nematode levels and plant damage. Common disease symptoms were leaf size reduction, yellowing and gradual decline of pomegranate plants, often observed in patches within orchards. Microscopic identification revealed distinctive pear-shaped body of mature females while J2 larvae displayed vermiform shapes and the associated species of M. incognita was confirmed through examination of the perineal pattern. Pathogenicity test reveals initiation of leaf yellowing symptom after 45 days of inoculation of larval suspension and root galling was observed after 60 days onward followed by plant decline under greenhouse conditions. Results from pot and field experiments demonstrated the efficacy of Fluopyram and Fluensulfone in reducing nematode populations and galling severity. Treatment with drenching of Fluopyram at the rate of 2 ml/L reduced 98.56 % larvae under field and 99.00 % larvae/100 cc soil under pot conditions. Statistical analysis (paired t-test and MANOVA) confirms significant differences in galling severity and larval population before and after drenching. The study also underscores the importance of weed management in disease mitigation as several weed species (Chenopodium album and Solanum nigrum) were identified as potential reservoirs for M. incognita in infested pomegranate plant basin. This investigation contributes to the advancement of management practices for pomegranate cultivation that addresses both nematode and weed infestations ultimately enhancing crop resilience and productivity.

Nanotherapeutic Approach to Delivery of Chemo- and Gene Therapy for Organ-Confined and Advanced Castration-Resistant Prostate Cancer.

Treatments for late-stage prostate cancer (CaP) have not been very successful. Frequently, advanced CaP progresses to castration-resistant prostate cancer (CRPC), with 50#37;-70% of patients developing bone metastases. CaP with bone metastasis-associated clinical complications and treatment resistance presents major clinical challenges. Recent advances in the formulation of clinically applicable nanoparticles (NPs) have attracted attention in the fields of medicine and pharmacology with applications to cancer and infectious and neurological diseases. NPs have been rendered biocompatible, pose little to no toxicity to healthy cells and tissues, and are engineered to carry large therapeutic payloads, including chemo- and genetic therapies. Additionally, if required, targeting specificity can be achieved by chemically coupling aptamers, unique peptide ligands, or monoclonal antibodies to the surface of NPs. Encapsulating toxic drugs within NPs and delivering them specifically to their cellular targets overcomes the problem of systemic toxicity. Encapsulating highly labile genetic therapeutics such as RNA within NPs provides a protective environment for the payload during parenteral administration. The loading efficiencies of NPs have been maximized while the controlled their therapeutic cargos has been released. Theranostic ("treat and see") NPs have developed combining therapy with imaging capabilities to provide real-time, image-guided monitoring of the delivery of their therapeutic payloads. All of these NP accomplishments have been applied to the nanotherapy of late-stage CaP, offering a new opportunity for a previously dismal prognosis. This article gives an update on current developments in the use of nanotechnology for treating late-stage, castration-resistant CaP.

Combination Modality Using Quercetin to Enhance the Efficacy of Docetaxel in Prostate Cancer Cells.

The standard of care chemotherapy drug presently used to treat castration-resistant prostate cancer (CRPC), docetaxel (Doc), also develops chemoresistance, thereby reducing its clinical utility. Since resistance to chemotherapy drugs can be overcome by co-treatment with plant-based bio-active compounds we undertook the present study to evaluate if quercetin (Que), a flavonoid present in plants such as onions, apples, olives, and grapes can enhance the efficacy of Doc. We studied the separate and combined effects of Que and Doc at different doses and different combination approaches in two different prostate cancer cell lines, DU-145 (moderately aggressive) and PC-3 (very aggressive), and assessed the effects of these combinations on viability, proliferation, and apoptosis. Monotherapy with these drugs showed dose-dependent cytotoxicity; however, only Doc monotherapy showed a statistically significant difference in IC50 levels (IC50 = 4.05 ± 0.52 nM for PC-3 and IC50 = 2.26 ± 0.22 nM for DU-145). In combination treatment, we used three different treatment approaches (TAP). The concentrations and range analyzed were chosen based on the approximate cytotoxicity of 30-50% when the drugs were used individually. Our observations indicate that the most beneficial effect of the Que and Doc combination was obtained with the TAP-2 approach, which is pre-treatment with all doses of Que for 24 h followed by low doses of Doc for another 24 h. Using this approach, we observed synergism at low concentrations of Doc (0.5 and 1.0 nM) and all concentrations of Que. An additive effect was observed at moderate and high concentrations of Doc (1.5, 2.0, and 2.5 nM) and all concentrations of Que in both cell lines. The TAP-2 strategy was also helpful in overcoming Doc resistance in resistant CaP cells. In summary, Que improved the therapeutic effect of Doc in CRPC, and it is proposed that this improvement is mediated through multiple mechanisms. This study provides a novel therapeutic modality for an effective combination using Doc and Que to enhance the efficacy of Doc in an innocuous manner for Doc resistance and CRPC treatment.

Use of Glycoproteins-Prostate-Specific Membrane Antigen and Galectin-3 as Primary Tumor Markers and Therapeutic Targets in the Management of Metastatic Prostate Cancer.

Galectins and prostate specific membrane antigen (PSMA) are glycoproteins that are functionally implicated in prostate cancer (CaP). We undertook this study to analyze the "PSMA-galectin pattern" of the human CaP microenvironment with the overarching goal of selecting novel-molecular targets for prognostic and therapeutic purposes. We examined CaP cells and biopsy samples representing different stages of the disease and found that PSMA, Gal-1, Gal-3, and Gal-8 are the most abundantly expressed glycoproteins. In contrast, other galectins such as Gal-2, 4-7, 9-13, were uniformly expressed at lower levels across all cell lines. However, biopsy samples showed markedly higher expression of PSMA, Gal-1 and Gal-3. Independently PSA and Gleason score at diagnosis correlated with the expression of PSMA, Gal-3. Additionally, the combined index of PSMA and Gal-3 expression positively correlated with Gleason score and was a better predictor of tumor aggressiveness. Together, our results recognize a tightly regulated "PSMA-galectin- pattern" that accompanies disease in CaP and highlight a major role for the combined PSMA and Gal-3 inhibitors along with standard chemotherapy for prostate cancer treatment. Inhibitor combination studies show enzalutamide (ENZ), 2-phosphonomethyl pentanedioic acid (2-PMPA), and GB1107 as highly cytotoxic for LNCaP and LNCaP-KD cells, while Docetaxel (DOC) + GB1107 show greater efficacy in PC-3 cells. Overall, 2-PMPA and GB1107 demonstrate synergistic cytotoxic effects with ENZ and DOC in various CaP cell lines.

Breast cancer prediction using a hybrid method based on Butterfly Optimization Algorithm and Ant Lion Optimizer.

The design and development of a computer-based system for breast cancer detection are largely reliant on feature selection techniques. These techniques are used to reduce the dimensionality of the feature space by removing irrelevant or redundant features from the original set. This article presents a hybrid feature selection method that is based on the Butterfly optimization algorithm (BOA) and the Ant Lion optimizer (ALO) to form a hybrid BOAALO method. The optimal subset of features chosen by BOAALO is utilized to predict the benign or malignant status of breast tissue using three classifiers: artificial neural network, adaptive neuro-fuzzy inference system, and support vector machine. The goodness of the proposed method is tested using 651 mammogram images. The results show that BOAALO outperforms the original BOA and ALO in terms of accuracy, sensitivity, specificity, kappa value, type-I, and type-II error as well as the receiver operating characteristics curve. Additionally, the suggested method's robustness is assessed and compared to five well-known methods using a benchmark dataset. The experimental findings demonstrate that BOAALO achieves a high degree of accuracy with a minimum number of features. These results support the suggested method's applicability for breast cancer diagnosis.

Surrogate model to screen for inactivation-based clearance of enveloped viruses during biotherapeutics process development.

Viral surrogates to screen for virus inactivation (VI) can be a faster, cheaper and safer alternative to third-party testing of pathogenic BSL2 (Biosafety level 2) model viruses. Although the bacteriophage surrogate, Ø6, has been used to assess low pH BSL2 VI, it has not been used for evaluation of detergent-mediated VI. Furthermore, Ø6 is typically assayed through host cell infectivity which introduces the risk of cross-contaminating other cell lines in the facility. To circumvent contamination, we developed an in-house RT-qPCR (Reverse transcriptase quantitative polymerase chain reaction) assay for selective detection of active Ø6 from a population of live and dead phage. The RT-qPCR assay was used to evaluate Ø6 inactivation in cell culture fluid of monoclonal antibody and fusion protein. Complementary Ø6 infectivity was also conducted at a third-party testing facility. The Ø6 RT-qPCR and infectivity data was modeled against VI of three BSL2 viruses, X- MuLV, A- MuLV and HSV-1 in corresponding therapeutics. Both Ø6 methods demonstrate that any VI agent showing Ø6 clearance of a minimum of 2.5 logs would demonstrate complete BSL2 VI of ≥ 4.0 logs. Compared to BSL2 virus testing, this in-house Ø6 RT-qPCR tool can screen VI agents at 5% the cost and a turnaround time of 2 to 3 days vs. 4 to 7 months.

Impact of nationwide lockdown on cancer care during COVID-19 pandemic: A retrospective analysis from western India.

BACKGROUND AND AIMS: COVID-19 has impacted healthcare system worldwide including cancer case. Aim of this study was to describe the experience of lockdown on cancer care concerning patient's visit and reception of treatment in western India. METHODS: This is a retrospective observational study conducted in patients with cancer attending a tertiary care center pre-lockdown and during lockdown (from January to May 2020). Data related to demographic parameters, type of tumor, type of treatment received and functional status of patients were retrieved from hospital medical records of patients. RESULTS: Of the 5258 patients included, 4363 visited hospital pre-lockdown (median age, 50 years) and 895 visited during the lockdown period (median age, 47 years). A total of 1168 and 106 patients visiting hospital before and during lockdown, respectively, had comorbidities. Breast cancer (25.6% and 29.7%), head and neck cancer (21.3% and 16.9%) were the most common type of solid tumors; leukemia (58.0% and 73.0%), lymphoma (18.8% and 13.5%) and multiple myeloma (18.6% and 12.2%) were the most common type of hematological malignancies observed in patients visiting pre-lockdown and during lockdown, respectively. Chemotherapy was most commonly received treatment (pre-lockdown, 71.8%; during lockdown, 45.9%). Other therapies reported includes supportive/palliative, targeted, hormonal, and immunotherapy. The majority of patients who visited the hospital pre-lockdown (68.4%) and during lockdown (62.8%) had 0 or 1 Eastern Cooperative Oncology Group (ECOG) score. CONCLUSION: Overall observations highlight a substantial impact of an imposed nationwide lockdown during COVID-19 pandemic on cancer care of patients in terms of reduced patient visits and number of treatments received.

Pediatric ocular motor cranial nerve palsy: Demographics and etiological profile.

Purpose: The aim of this study was to describe epidemiological and clinical characteristics of pediatric ocular motor cranial nerve palsy. Methods: This was a retrospective record-based study, carried out at a tertiary eye care hospital in India, between January 2011 and January 2015 and included patients up to 16 years of age at the time of presentation, diagnosed with third, fourth, sixth nerve palsy or a combination of these with other cranial nerve palsy. Data analyzed included demographic details, etiologies, presence or absence of amblyopia, relevant investigations, and management. Results: A total of 90 cases were included in the study. Eighty patients (88.88%) presented with isolated nerve palsy. Forty-three patients (47.77%) had congenital nerve palsy. The most common nerve involved was third (n = 35, 38.88%) followed by sixth (n = 23, 25.55%) and fourth nerve (n = 22, 24.44%). Most common cause of third and fourth cranial nerve palsy was congenital (n = 18, 51.42% and n = 17, 77.30%, respectively), while it was trauma for the sixth nerve (n = 7, 30.40%). Amblyopia was most frequently associated with third cranial nerve palsy (n = 27, 77.14%). The radio-imaging yield was maximum (n = 7, 70%) for combined cranial nerve palsy. Overall 44 (48.88%) patients were managed conservatively, while 46 (51.11%) patients needed squint with or without ptosis surgery. Conclusion: The most common ocular motor cranial nerve involved in the pediatric population was the third cranial nerve, and it was found to be the most amblyogenic in this age group. The neuroimaging yield was maximum for combined cranial nerve palsy. The most common conservatively managed nerve palsy in this study group was the fourth nerve palsy.

Mitomycin-C in Post-endoscopic Septoplasty Synechiae: Does it Really Helps?

Endoscopic septoplasty has advantage of magnified image and access to posterior part of the septum. Synechiae formation is a common complication of this procedure. The incidence of adhesions rises particularly in cases of submucosal flap tears which is sometimes unavoidable during spurectomy. Various procedures like septal splints, nasal packings have been tried to reduce the incidence of adhesions. Recently mitomycin-C is being tried to decrease post operative adhesions after Functional Endoscopic Sinus Surgery, endoscopic Dacrocystorhinostomy, with varying degree of success. Mitomycin-C has got anti fibroblastic activity, which inhibits the fibroblasts without hampering epithelization. Eighty (80) Patients undergoing septoplasty and spurectomy were observed for any submucosal flap tear, and those developing flap tear were included in the study. The patients were divided into 2 groups of 40 each. In group-1 topical mitomycin (0.4 mg/ml) was applied at the raw area for 5 min, similarly normal saline was applied in group-2 (control).The aim of study was to evaluate the role of mitomycin-C in preventing post operative adhesions in Endoscopic septoplasty. Adhesion rate was calculated at 1 month, and compared using Chi square test. P value < 0.05 was considered significant. The adhesion rate at 1 month was 17.5% in group-1 and 45% in group-2 (P = 0.008). Incidence of adhesion decreased significantly in mitomycin-C treated group. Mitomycin-C can routinely be used in septoplasty to prevent post op adhesions, particularly in cases of submucosal flap tears/concurrent turbinate reduction procedures.

Intra-articular Migration of Broken Patellar Tension Band Wire: A Rare Case.

INTRODUCTION: Comminuted fracture of patella is most commonly treated by tension band wiring with cerclage wiring. Broken cerclage wiring after such surgery is not uncommon but intra-articular migration of broken cerclage wire is rarely known and described complication. CASE REPORT: A 51-year-old male patient of Indian origin which was treated, 15 months back for fracture patella with tension band wiring with cerclage wiring, presented with painful restriction of knee movements for the last 4 weeks. X-ray showed cerclage wire had broken at multiple sites with one broken piece migrated intra-articular. This case is unique due to the size of migrated broken cerclage wire which was about 4.5 cm long, second the anterior part of broken wire was hooked around anterior cruciate ligament, which prevented its further migration otherwise neurovascular structures could have been damaged. This intra-articular piece of wire was removed arthroscopically. CONCLUSIONS: We recommend that all patients receiving bone fixation with wires should be counseled about the risks of hardware failure and possible migration, with the option of elective hardware removal discussed. In addition, when wire breakage is detected radiologically, the patient should be strongly advised to have the wires removed as soon as possible.

Molecular Detection of Foodborne Pathogens: A Rapid and Accurate Answer to Food Safety.

Food safety is a global health concern. For the prevention and recognition of problems related to health and safety, detection of foodborne pathogen is of utmost importance at all levels of food production chain. For several decades, a lot of research has been targeted at the development of rapid methodology as reducing the time needed to complete pathogen detection tests has been the primary goal of food microbiologists. With the result, food microbiology laboratories now have a wide array of detection methods and automated technologies such as enzyme immunoassay, polymerase chain reaction, and microarrays, which can cut test times considerably. Nucleic acid amplification strategies and advances in amplicon detection methodologies have been the key factors in the progress of molecular microbiology. A comprehensive literature survey has been carried out to give an overview in the field of foodborne pathogen detection. In this paper, we describe the conventional methods, as well as recent developments in food pathogen detection, identification, and quantification, with a major emphasis on molecular detection methods.

Improvement of efficiency of oil extraction from wild apricot kernels by using enzymes.

An experiment was conducted to evaluate and standardize the protocol for enhancing recovery of oil and quality from cold pressed wild apricot kernels by using various enzymes. Wild apricot kernels were ground into powder in a grinder. Different lots of 3 kg powdered kernel were prepared and treated with different concentrations of enzyme solutions viz. Pectazyme (Pectinase), Mashzyme (Cellulase) and Pectazyme + Mashzyme. Kernel powder mixed with enzyme solutions were kept for 2 h at 50(±2) °C temperature for enzymatic treatment before its use for oil extraction through oil expeller. Results indicate that use of enzymes resulted in enhancement of oil recovery by 9.00-14.22 %. Maximum oil recovery was observed at 0.3-0.4 % enzyme concentration for both the enzymes individually, as well as in combination. All the three enzymatic treatments resulted in increasing oil yield. However, with 0.3 % (Pectazyme + Mashzyme) combination, maximum oil recovery of 47.33 % could be observed against were 33.11 % in control. The oil content left (wasted) in the cake and residue were reduced from 11.67 and 11.60 % to 7.31 and 2.72 % respectively, thus showing a high increase in efficiency of oil recovery from wild apricot kernels. Quality characteristics indicate that the oil quality was not adversely affected by enzymatic treatment. It was concluded treatment of powdered wild apricot kernels with 0.3 % (Pectazyme + Mashzyme) combination was highly effective in increasing oil recovery by 14.22 % without adversely affecting the quality and thus may be commercially used by the industry for reducing wastage of highly precious oil in the cake.

Sclerosing stromal tumor of the ovary.

Sclerosing stromal tumor is a rare ovarian tumor, occurring in young adults in the second and third decade of life. We report clinical and histopathological features of three cases of sclerosing stromal tumor of the ovary with a review of literature. The tumor has distinct histological features and is easily recognizable when a high index of suspicion is maintained in young patients presenting with an ovarian mass. These tumors are benign and can be treated successfully by enucleation or unilateral ovariotomy.

Reduction of non-enzymatic browning of orange juice and semi-concentrates by removal of reaction substrate.

A study was conducted to standardize the technology for the removal of amino acids (one of the browning reaction substrates) from sweet orange cv. Malta Common juice to reduce colour and quality deterioration in single strength juice and during subsequent concentration. Juice of sweet orange (Citrus sinensis) cv. Malta Common fruits was extracted by screw type juice extractor, preserved in 500 ppm SO2 and clarified by using "Pectinase CCM" enzyme (0.2% for 2 h at 50 ± 2 °C). For removal of amino acids juice was passed under gravity through a glass column packed with an acidic cation exchange resin (CER), Dowex-50 W and quantity to be treated in one lot was standardized. The CER treated and untreated juices were concentrated to 15 and 30°Brix in a rotary vacuum evaporator. Results indicate that 121 ml of orange juice when passed through a glass column (5 cm internal diameter) packed with cation exchange resin (Dowex-50 W) upto a height of 8 cm, could remove about 98.4% of the amino acids with minimum losses in other juice constituents. With cation exchange resin treatment, the non-enzymatic browning and colour deterioration of orange juice semi-concentrates was reduced to about 3 folds in comparison to untreated counterparts. The retention of vitamin C and sugars was also better in semi-concentrates prepared from cation exchange resin treated juice. Thus, cation exchange resin treatment of orange juice prior to concentration and storage is highly beneficial in reduction of non-enzymatic browning, colour deterioration and retention of nutritional, sensory quality of product during preparation and storage.

Primary squamous cell carcinoma of the breast.

Primary squamous cell carcinoma (SqCC) of the breast is a very rare tumor accounting for less than 0.1% of all invasive breast carcinomas. This is a very aggressive, hormone receptor negative and treatment refractory tumor with poor prognosis. We report a case of primary SqCC of the breast in a 72-year-old postmenopausal lady who presented with a lump in the right breast. The treatment of primary SqCC of the breast does not differ from other common histological types of breast cancer and may involve surgery, chemotherapy, hormonal therapy and radiation therapy. Clinical trials including large series of these rare tumors are needed to increase our knowledge and to improve patient's outcome.

Spectrum of male breast lesions diagnosed by fine needle aspiration cytology: a 5-year experience at a tertiary care rural hospital in central India.

This study examines the spectrum of lesions in the male breast at a tertiary care rural hospital in central India and explores the role of Fine Needle Aspiration Cytology (FNAC) in the diagnosis of these lesions. Male breast lesions formed 5.89% (119 cases) of the 2017 breast lesions which were sent for FNA in the cytology section over a period of 5 years (January 2005-December 2009). Of these, biopsy had been performed only on 37 (31.1%) patients. Benign lesions comprised 102 (85.7%) cases, malignant lesions comprised 14 (11.8%) cases and inflammation/abscess was found in 3 (2.5%) cases. Gynecomastia was the commonest benign lesion in 86 (84.3%) cases. The cytologic features of gynecomastia included mild to moderate cellularity, cohesive sheets of bland cells, bipolar bare nuclei. Mild nuclear atypia was found in 19 cases. The cytologic features of malignancy comprised of dyshesive groups of ductular cells with moderate to severe degree of nuclear atypia and absence of bare nuclei. Histology was done in 37 cases and diagnostic accuracy of FNAC for gynecomastia was 100% and for malignancy 85.7%. This study showed that FNAC is a reliable tool for diagnosing male breast lesions. We conclude that FNAC should be performed as a standard procedure in the clinical evaluation of male breast masses. Many unnecessary surgical biopsies for histopathologic diagnosis can thus be avoided.

A plant-based system for rapid production of influenza vaccine antigens.

BACKGROUND: Influenza virus is a globally important respiratory pathogen that causes a high degree of annual morbidity and mortality. Significant antigenic drift results in emergence of new, potentially pandemic, virus variants. The best prophylactic option for controlling emerging virus strains is to manufacture and administer pandemic vaccines in sufficient quantities and to do so in a timely manner without impacting the regular seasonal influenza vaccine capacity. Current, egg-based, influenza vaccine production is well established and provides an effective product, but has limited capacity and speed. OBJECTIVES: To satisfy the additional global demand for emerging influenza vaccines, high-performance cost-effective technologies need to be developed. Plants have a potential as an economic and efficient large-scale production platform for vaccine antigens. METHODS: In this study, a plant virus-based transient expression system was used to produce hemagglutinin (HA) proteins from the three vaccine strains used during the 2008-2009 influenza season, A/Brisbane/59/07 (H1N1), A/Brisbane/10/07 (H3N2), and B/Florida/4/06, as well as from the recently emerged novel H1N1 influenza A virus, A/California/04/09. RESULTS: The recombinant plant-based HA proteins were engineered and produced in Nicotiana benthamiana plants within 2 months of obtaining the genetic sequences specific to each virus strain. These antigens expressed at the rate of 400-1300 mg/kg of fresh leaf tissue, with >70% solubility. Immunization of mice with these HA antigens induced serum anti-HA IgG and hemagglutination inhibition antibody responses at the levels considered protective against these virus infections. CONCLUSIONS: These results demonstrate the feasibility of our transient plant expression system for the rapid production of influenza vaccine antigens.

Hashimoto's disease in a bilateral benign cystic ovarian teratoma: a case report.

A wide array of tissues derived from all the three germinal layers is seen in ovarian teratomas. Among these, thyroid tissue is present in 10% cases of all mature cystic teratomas. We report this case of Hashimoto's thyroiditis in a clinically euthyroid patient who tested positive for antithyroid peroxidase antibodies in spite of normal thyroid hormone profile. While the histological features of several disorders of thyroid tissue may be discovered, Hashimoto's thyroiditis is extremely rare finding in ovarian teratomas.

Prostate-derived Ets transcription factor (PDEF) is a potential prognostic marker in patients with prostate cancer.

BACKGROUND: Reduced expression of prostate-derived Ets transcription factor (PDEF) leads to morphologic change as well as increased migration and invasiveness of prostate cancer cells. However, the clinical relevance of PDEF expression and its relationship to anti-apoptotic protein survivin is yet to be determined. METHODS: Tissue microarrays of 73 prostate carcinomas and their adjacent benign prostate tissue, as well as 50 benign prostates were evaluated for PDEF expression by immunohistochemistry. Results were confirmed in available tumor tissues using Western blot and RT-PCR. Expression of survivin in prostate carcinoma and benign tissues were determined using Western blot. Results and correlation with clinical data were statistically analyzed. RESULTS: Patients' specimens with low Gleason scores (GS < 5) expressed higher levels of PDEF protein and lower levels of survivin protein when compared with moderate-to-high GS tumors (GS > 6). Patients with PDEF-positive tumor survived significantly longer (P < 0.0001) than patients with PDEF-negative tumor, and the 8-year survival rate was 94% and 40%, respectively. PDEF expression was detected at the highest levels in benign tissues and was down-regulated or lost in 30 recently diagnosed prostate carcinomas. Re-expression of PDEF in prostate cancer cells inhibited survivin expression. Treatment of prostate cancer cells with methylseleninic acid resulted in restoration of PDEF expression, down-regulation of survivin, and inhibition of tumor cell growth when compared with untreated controls (P < 0.05). CONCLUSIONS: These studies demonstrated an inverse correlation between PDEF and survivin expression, and that up-regulation of PDEF was associated with a favorable prognosis in patients with clinically localized prostate cancer.

Plant-based rapid production of recombinant subunit hemagglutinin vaccines targeting H1N1 and H5N1 influenza.

In 2009, a novel H1N1 swine influenza virus was isolated from infected humans in Mexico and the United States, and rapidly spread around the world. Another virus, a highly pathogenic avian influenza virus of the H5N1 subtype, identified by the World Health Organization as a potential pandemic threat in 1997, continues to be a significant risk. While vaccination is the preferred strategy for the prevention and control of influenza infections, the traditional egg-based approach to producing influenza vaccines does not provide sufficient capacity and adequate speed to satisfy global needs to combat newly emerging strains, seasonal or potentially pandemic. Significant efforts are underway to develop and implement new cell substrates with improved efficiency for influenza vaccine development and manufacturing. In recent years, plants have been used to produce recombinant proteins including subunit vaccines and antibodies. The main advantages of using plant systems for the production of vaccine antigens against influenza are their independence from pathogenic viruses, and cost and time efficiency. Here, we describe the large-scale production of recombinant hemagglutinin proteins from A/California/04/09 (H1N1) and A/Indonesia/05/05 (H5N1) strains of influenza virus in Nicotiana benthamiana plants, and their immunogenicity (serum hemagglutination inhibition and virus neutralizing antibodies), and safety in animal models. These results support the testing of these candidate vaccines in human volunteers and also the utility of our plant expression system for large-scale recombinant influenza vaccine production.