Overexpression of native ORANGE (OR) and OR mutant protein in Chlamydomonas reinhardtii enhances carotenoid and ABA accumulation and increases resistance to abiotic stress.

The carotenoid content of plants can be increased by overexpression of the regulatory protein ORANGE (OR) or a mutant variant known as the 'golden SNP'. In the present study, a strong light-inducible promoter was used to overexpress either wild type CrOR (CrORWT) or a mutated CrOR (CrORHis) containing a single histidine substitution for a conserved arginine in the microalgae Chlamydomonas reinhardtii. Overexpression of CrORWT and CrORHis roughly doubled and tripled, respectively, the accumulation of several different carotenoids, including ß-carotene, α-carotene, lutein and violaxanthin in C. reinhardtii and upregulated the transcript abundance of nearly all relevant carotenoid biosynthetic genes. In addition, microscopic analysis revealed that the OR transgenic cells were larger than control cells and exhibited larger chloroplasts with a disrupted morphology. Moreover, both CrORWT and CrORHis cell lines showed increased tolerance to salt and paraquat stress. The levels of endogenous phytohormone abscisic acid (ABA) were also increased in CrORWT and CrORHis lines, not only in normal growth conditions but also in growth medium supplemented with salt and paraquat. Together these results offer new insights regarding the role of the native OR protein in regulating carotenoid biosynthesis and the accumulation of several carotenoids in microalgae, and establish a new functional role for OR to modulate oxidative stress tolerance potentially mediated by ABA.

Ectopic expression of ORANGE promotes carotenoid accumulation and fruit development in tomato.

Carotenoids are critically important to plants and humans. The ORANGE (OR) gene is a key regulator for carotenoid accumulation, but its physiological roles in crops remain elusive. In this study, we generated transgenic tomato ectopically overexpressing the Arabidopsis wild-type OR (AtORWT ) and a 'golden SNP'-containing OR (AtORHis ). We found that AtORHis initiated chromoplast formation in very young fruit and stimulated carotenoid accumulation at all fruit developmental stages, uncoupled from other ripening activities. The elevated levels of carotenoids in the AtOR lines were distributed in the same subplastidial fractions as in wild-type tomato, indicating an adaptive response of plastids to sequester the increased carotenoids. Microscopic analysis revealed that the plastid sizes were increased in both AtORWT and AtORHis lines at early fruit developmental stages. Moreover, AtOR overexpression promoted early flowering, fruit set and seed production. Ethylene production and the expression of ripening-associated genes were also significantly increased in the AtOR transgenic fruit at ripening stages. RNA-Seq transcriptomic profiling highlighted the primary effects of OR overexpression on the genes in the processes related to RNA, protein and signalling in tomato fruit. Taken together, these results expand our understanding of OR in mediating carotenoid accumulation in plants and suggest additional roles of OR in affecting plastid size as well as flower and fruit development, thus making OR a target gene not only for nutritional biofortification of agricultural products but also for alteration of horticultural traits.

Salvage of the thiamin pyrimidine moiety by plant TenA proteins lacking an active-site cysteine.

The TenA protein family occurs in prokaryotes, plants and fungi; it has two subfamilies, one (TenA_C) having an active-site cysteine, the other (TenA_E) not. TenA_C proteins participate in thiamin salvage by hydrolysing the thiamin breakdown product amino-HMP (4-amino-5-aminomethyl-2-methylpyrimidine) to HMP (4-amino-5-hydroxymethyl-2-methylpyrimidine); the function of TenA_E proteins is unknown. Comparative analysis of prokaryote and plant genomes predicted that (i) TenA_E has a salvage role similar to, but not identical with, that of TenA_C and (ii) that TenA_E and TenA_C also have non-salvage roles since they occur in organisms that cannot make thiamin. Recombinant Arabidopsis and maize TenA_E proteins (At3g16990, GRMZM2G080501) hydrolysed amino-HMP to HMP and, far more actively, hydrolysed the N-formyl derivative of amino-HMP to amino-HMP. Ablating the At3g16990 gene in a line with a null mutation in the HMP biosynthesis gene ThiC prevented its rescue by amino-HMP. Ablating At3g16990 in the wild-type increased sensitivity to paraquat-induced oxidative stress; HMP overcame this increased sensitivity. Furthermore, the expression of TenA_E and ThiC genes in Arabidopsis and maize was inversely correlated. These results indicate that TenA_E proteins mediate amidohydrolase and aminohydrolase steps in the salvage of thiamin breakdown products. As such products can be toxic, TenA_E proteins may also pre-empt toxicity.

Association between neutrophil geletinase-associated lipocalin and iron deficiency anemia in children on chronic dialysis.

BACKGROUND: Iron deficiency anemia (IDA) in children with chronic kidney disease (CKD) is common and associated with higher risk of death. Neutrophil geletinase-associated lipocalin (NGAL) is a small 25 kDa glycoprotein, a member of lipocalin superfamily that released at the response of cellular stress from different cells. In addition, NGAL was studied as an iron regulatory glycoprotein and regulator of iron related gene. The aim of the current study was to determine any association between serum NGAL and body iron status markers in children on chronic dialysis. MATERIALS AND METHODS: This correlation study was carried out between May 2012 and May 2013 and evaluated all dialysis patients less than 19 years in pediatric dialysis centers in Isfahan that didn't have exclusion criteria. They were 40 children, including 23 persons on hemodialysis (HD) and 17 persons dialyzed by peritoneal dialysis (PD). Furthermore, we selected 40 children as healthy controls. We examined the relationship between plasma NGAL levels and indices of anemia such as ferritin, transferrin saturation (TSAT) and serum iron (SI) in dialysis children. RESULTS: Serum NGAL level in children on chronic dialysis (group including both PD and HD patients) was significantly higher than healthy controls (P = 0.008). Furthermore, in this group Serum NGAL level had inverse correlation with TSAT (P = 0.04, r = -0.22), SI (P = 0.04, r = -0.2), white blood cells (P = 0.045, r = -0.26) and serum ferritin (P = 0.006, r = -0.3). In addition, HD patients had higher serum NGAL level than PD patients (P = 0.048). CONCLUSION: High serum NGAL level in low TSAT group demonstrated that NGAL probably has an important role in IDA in children on chronic dialysis; therefore, it can be a new marker for diagnosis of IDA in CKD.

Concomitant surgical correction of severe stress urinary incontinence and anterior vaginal wall prolapse by anterior vaginal wall wrap: 18 months outcomes.

BACKGROUND: The aim of this study is to evaluate the outcome of an innovative, minimally invasive sling technique with autologous tissue in women with concomitant incontinence and anterior vaginal wall prolapse (AVWP). MATERIALS AND METHODS: Fifty-six women with stress urinary incontinence (SUI) or mixed urinary incontinence and AVWP were randomly assigned into two groups: In Group A (26 patients), anterior colporrhaphy (Kelly placation) and sling placement using a strip of anterior vaginal wall were performed, and in Group B (30 patients), transvaginal mesh correction of AVWP and tension-free vaginal tape (TVT) insertion (retropubic - craniocaudal route) using polypropylene mesh were carried out. The patients were followed-up for over 18 months and were assessed objectively using a 48 h frequency-volume chart, a 48 h pad test and a standardized stress test. Related surgical complications and outcomes were recorded and compared. RESULTS: Surgical cure rates for Group A and Group B at the first (3 days) and last (18 months) post-operative visits were 62% and 84%; and 54%, and 72%, respectively (P = 0.09 and 0.31). Complications occurred in 9 patients (44%) of Group B, but only 3 patients (12%) in Group A. CONCLUSION: Vaginal sling surgery using an anterior vaginal wall strip can improve SUI and in comparison with propylene mesh is associated with lower complication rates. Although, the surgical success rate of this technique is lower than T-Sling, larger studies with selected patients will help assess the suitable patients for this pelvic reconstructive surgery.

Diagnostic value of total serum/free prostate specific antigen and prostate cancer antigen-3 levels in prostate cancer.

BACKGROUND: The purpose of this study was to compare serum total and free prostate specific antigen (PSA) levels and serum prostate cancer antigen-3 (PCA3) levels in patients with prostate cancer in 2018 and 2019. METHODS: This research was a prospective case-control study. The case group included all patients with suspected prostate cancer, and the control group included individuals without prostate disease who were referred to Ali Asghar and Nour Hospital in Isfahan, Iran, from October 2018 to October 2020. The serum total PSA, free PSA, and PCA3 levels in both groups were measured using the ELISA method with standard kits and compared between the groups. RESULTS: The two groups were matched in terms of age and body mass index (BMI). The results showed that the mean free PSA level in the control group was significantly higher than that in the case group (P<0.05). Conversely, the mean total PSA level in the case group was significantly higher than that in the control group (P<0.05). However, no significant difference was observed in the mean PCA3 levels between the case and control groups. In addition, the total PSA variable with a cutoff of ≤3.14 exhibited 93% sensitivity and 82% specificity, demonstrating the highest diagnostic accuracy in distinguishing between prostate cancer and healthy individuals. Similarly, the PCA3 value with a cutoff of ≤3.5 had a sensitivity and specificity of 70% and 72%, respectively. CONCLUSION: Overall, the study results indicated that total PSA and PCA3 levels have higher diagnostic accuracy in distinguishing patients with suspected prostate cancer from healthy individuals.

Pericardial effusion among children: Retrospective analysis of the etiology and short-term outcome in a referral center in the south of Iran.

Background and Aims: We reinvestigated the causes, symptoms, and management of childhood pericardial effusion (PE) and its gradual changes during recent years in a referral pediatric cardiology center in the south of Iran. Methods: We retrospectively analyzed the profile of PE patients who were under 18 years old from 2015 to 2020. The patient's demographic, clinical, and paraclinical information was extracted and analyzed using SPSS software. Result: In general, 150 out of 63,736 admitted patients (0.23% of the total pediatric admissions) were diagnosed with PE (male/female 1:1.17). The median age was 3.25 years (range:\ 2 days to 18 years; interquartile range: 9.5), and 50% of them were under 3 years of age. 32.6% had moderate to severe PE. Most patients presented with acute symptoms (68%) and respiratory problems, as the most common symptoms (30.6%). Tamponade signs were presented in 2% (n = 3) of the patients, and 80.7% (n = 121) were in a stable hemodynamic condition. In total, renal failure (22%) and parapneumonic effusion were the leading etiologies. Viral (7%) and bacterial (5%) pericarditis were the seventh and eighth causes; however, in severe cases, renal failure (22%) and bacterial pericarditis (14%) were dominant. In total, 14.1% (n = 21) of the patients needed pericardiocentesis that increased to 78.3% (n = 18) in severe cases. Only 6% had persistent PE for more than 3 months. Conclusion: Childhood PE is mostly a result of renal failure and noninfectious causes. True pericarditis cases are not common, except in severe cases. It is more common in less than 3-year-old patients, and chronicity is rare. Severe cases had a high chance of pericardiocentesis, but other cases were mainly managed by treatment of the underlying causes.

Reducing the risk of death induced by aluminum phosphide poisoning: The new therapies.

Numerous people suffer from accidental or deliberate exposure to different pesticides when poisoning with aluminum phosphate (AlP) is increasing in the eastern countries. Aluminum phosphate is a conventional insecticide that quickly reacts with water or the moistures in the atmosphere and produces fatal phosphine gas, which absorbs quickly by the body. Oral consumption or inhalation of AlP leads to excessive reaction of the body such as fatigue, vomiting, fever, palpitation, vasodilatory shock, increasing blood pressure, cardiac dysfunction, pulmonary congestion, shortness of breath, and death. The garlic smell from the patient's mouth or exhale is one of the methods to recognize the positioning. Due to the lack of individual antidotes, several supportive treatments are required. The present study focused on the available and new therapies that help reduce the effect of AlP poisoning and the mortality rate. The therapies are divided into the antioxidant-related agent and the other agents. The impacts of each agent on the experimental cases are reported.

Inflammatory and T Helper 17/ Regulatory T Cells Related Cytokines Balance in Cutaneous Lupus Erythematosus (CLE).

The cutaneous lupus erythematosus (CLE) is a common manifestation among systemic lupus erythematosus (SLE) patients. Malar rash and discoid lupus (DLE) are in the category of acute and chronic CLE, respectively. The pathogenesis of CLE is multifactorial, and cytokine imbalances contribute to immune dysfunction and the induction of organ damage. Many aspects of cytokine dysregulation are still unclear in SLE and in particular CLE. Therefore, we concurrently measured the inflammatory [Tumor necrosis factor-alpha (TNF-α) and Interleukin (IL)-6)], T helper (Th)-17 (IL-17 and IL-23) and regulatory T cells [Transforming growth factor-beta (TGFß) and IL-10)]-related cytokines in patients with CLE (patients with malar rash and/or DLE) and compared them with SLE patients and healthy individuals (n=25 in each group, a total of 75 patients). The serum levels of cytokines were assessed by Enzyme-Linked Immunosorbent Assay (ELISA) method. IL-6 cytokine was significantly higher in SLE, DLE, and malar rash patients compared to those in healthy controls (p=0.025) and in patients with arthralgia (p=0.038), and gastrointestinal involvement (p=0.048). IL-17 was significantly higher in malar rash patients compared to normal individuals (p=0.023), SLE (p=0.008) and DLE patients (p=0.019) and in patients with oropharyngeal ulcer (p=0.05) but, IL-23 was significantly higher only in DLE patients than healthy controls (p=0.019). In conclusion, inflammatory cytokines such as IL-6 involved in inflammation and differentiation of Th17 cells are probably responsible in part for Th17 activity in CLE. IL-17, IL-23, and IL-6/IL-6R (IL-6 receptor) inhibitors may be good treatments for CLE patients. So targeting these cytokines activity pathways can improve the CLE treatment strategy and may open a novel guideline for SLE and CLE treatment.

Predominant role of innate pro-inflammatory cytokines in vitiligo disease.

Vitiligo is a skin disorder with melanocyte destruction and an autoimmune basis. Given the importance of cytokines in autoimmunity, we aimed to find the cytokine profile of innate and adaptive immunity in vitiligo patients, and correlate them with clinical parameters. The serum levels of innate immunity [interleukin(IL)-1α, IL-1ß, IL-6, IL-8, IL-12, IL-15 and tumor necrosis factor (TNF)-α] and T helper(Th)1 [IL-2, interferon (IFN)-γ, TNF-ß], Th2 (IL-4, IL-5, IL-10, IL-13) and Th17 (IL-17, IL-23) cytokines in 44 vitiligo patients were measured by multiplex cytokine assay and compared with 44 healthy subjects. All innate immunity (p < 0.04), Th1 (p < 0.01), Th2 (p < 0.05) and Th17 (p < 0.001) cytokines were higher in patients than controls. Total summation levels of innate immunity and adaptive immunity cytokines showed a remarkable up-regulation in the patients (p < 0.0001). The ratio of innate immunity to Th1 (p = 0.03), Th2 (p = 0.01) and Th17 (p = 0.03) cytokines was significantly higher in patients vs. controls. We found significant higher ratio of Th1 to Th2 cytokines and TNF-ß elevated levels in patients with a family history of autoimmunity (p < 0.05). IL-4 and IL-13 (p < 0.04) levels were lower in patients with amelanotic hair. Increased IL-10 level was observed in patients with stable disease (p = 0.02).In conclusion, the profile of cytokines in patients showed a dominant role of innate immunity pro-inflammatory cytokines in vitiligo, which suggests the potential of targeting these cytokines for vitiligo treatment. While a higher ratio of Th1/Th2 cytokines was observed in the patients, association of decreased Th2 cytokines with disease complications suggests a protective role for Th2 pathway.

An array microhabitat device with dual gradients revealed synergistic roles of nitrogen and phosphorous in the growth of microalgae.

Harmful algal blooms (HABs) are an emerging environmental problem contaminating water resources and disrupting the balance of the ecosystems. HABs are caused by the sudden growth of photosynthetic algal cells in both fresh and marine water, and have been expanding in extent and appearing more frequently due to the climate change and population growth. Despite the urgency of the problem, the exact environmental conditions that trigger HABs are unknown. This is in part due to the lack of high throughput tools for screening environmental parameters in promoting the growth of photosynthetic microorganisms. In this article, we developed an array microhabitat device with well defined dual nutrient gradients suitable for quantitative studies of multiple environmental parameters in microalgal cell growth. This device enabled an ability to provide 64 different nutrient conditions [nitrogen (N), phosphorous (P), and N : P ratio] at the same time, and the gradient generation took less than 90 min, advancing the current pond and test tube assays in terms of time and cost. Using a photosynthetic algal cell line, Chlamydomonas reinhardtii, preconditioned in co-limited media, we revealed that N and P synergistically promoted cell growth. Interestingly, no discernible response was observed when single P or N gradient was imposed. Our work demonstrated the enabling capability of the microfluidic platform for screening effects of multiple environmental factors in photosynthetic cell growth, and highlighted the importance of the synergistic roles of environmental factors in algal cell growth.

Serum levels of total and urine level of PCA3 in patients with benign prostatic hyperplasia and prostate cancer.

BACKGROUND: Urine test with the PSA result will provide a good prognosis of the prostate cancer. Therefore, considering the importance of PCA3 in this study, we aimed to compare the serum total and urinary PCA3 levels in patients with benign hyperplasia and prostate cancer. METHODS: This cross-sectional study was performed on 90 patients referring to Noor and Hazrat-e-Ali Asghar Hospital in Isfahan from October 2017 to October 2018 for prostate biopsy. Patients were divided into two groups including benign prostate hyperplasia (BPH) and prostate cancer. Serum total and urinary PCA3 levels were measured and compared in both groups. RESULTS: 38 patients with prostate cancer and 52 patients with BPH participated in this study. Mean age in prostate cancer group was significantly higher than BPH group (P=0.01). Also mean PCA3, and total PSA, in patients with prostate cancer was significantly higher than patients with BPH (P<0.05). CONCLUSION: PCA3 was an important marker in patients with prostate cancer and BPH.

Interleukin-27 gene variant rs153109 is associated with enhanced cytokine serum levels and susceptibility to Behçet's disease in the Iranian population.

Behcet's disease (BD) is a systemic vasculitis, characterized by recurrent oral aphthous, genital ulcers, ocular lesions, and other organ involvement. Interleukin (IL)-27 with its pro- and anti-inflammatory effects might be an important effective cytokine in this disease. The aim of this study was to investigate the association of IL-27 serum concentration and a single-nucleotide polymorphism (SNP) rs153109 (-964 A > G) with the risk and clinical features of the patients with BD. IL-27 Genotyping was performed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and the IL-27 serum levels were measured using enzyme-linked immunosorbent assay (ELISA). It is shown that AG, GG, and AG + GG genotypes, as well as G allele of rs153109, can significantly increase the risk of BD in total and in male individuals. Significantly higher frequencies of AG and GG genotypes and G allele were observed in total and male patients with an active form of BD. AG and GG genotypes were associated with joint (p = 0.046) and vascular (p = 0.02) involvement. The frequency of the G allele was higher in all patients, as well as in female patients with vascular involvement (p = 0.02). Serum cytokine analysis indicated an increased level of IL-27 in BD patients compared to healthy subjects (p = 0.038). Additionally, a higher level of IL-27 was detected in patients carrying the rs153109 GG genotype (p = 0.04) and those with renal (p = 0.009) and skin (p = 0.05) involvement. In conclusion, this study underscores the involvement of IL-27 rs153109 variants and increased serum level in BD susceptibility and pathogenesis.

Interleukin-17 production by CD4+CD45RO+Foxp3+ T cells in peripheral blood of patients with atherosclerosis.

INTRODUCTION: T regulatory cells (Tregs) are known as immunoregulatory cells that are reduced in atherosclerosis. Tregs are a part of crosstalk between the immune system and lipoprotein metabolism, both of which are involved in atherosclerotic processes. Depletion of Tregs leads to impaired clearance of low density lipoprotein (LDL), and intracellular cholesterol homeostasis affects Treg cell development. Furthermore, the atherosclerotic environment affects the Treg cells' phenotype and plasticity. Plasticity between Tregs and Th17 cells has been a matter of investigation lately. We investigated the frequency of interleukin-17 (IL-17)-producing Tregs in the peripheral blood of patients with atherosclerosis. MATERIAL AND METHODS: We studied 10 non-diabetic patients with significant coronary artery disease (CAD) as the patient group, and seven non-diabetic individuals with normal coronary angiography/insignificant CAD as the control group. Peripheral blood mononuclear cells were stained with fluorescent antibodies to detect CD4, CD45RO, IL-17, and Foxp3 expression both before and after stimulation with PMA/Ionomycin. Cell enumeration was performed using flowcytometry and analysed using Mann-Whitney test. RESULTS: CD4+IL-17+Foxp3+ and CD4+IL-17+Foxp3- subsets showed higher frequencies in patients than in controls both before (p = 0.0031, p = 0.033, respectively) and after stimulation (p = 0.0027 and p = 0.0013, respectively). Interestingly, CD4+IL-17+Foxp3+ cells were almost exclusively CD45RO+ with a much higher frequency in patients than in controls (p = 0.0027, p = 0.0007). After stimulation, the frequency of CD4+CD45RO+IL-17+Foxp3+ lymphocytes increased to a greater extent in patients (p < 0.0001) than in controls. CONCLUSIONS: Interleukin-17 production by an intermediate population with an activated Treg phenotype in our patients may point to the population heterogeneity or plasticity in Tregs during atherosclerotic inflammation.

Can the kidney volume help to differentiate the types of rejection before biopsy?

BACKGROUND: The aim of this study was to use the volume of the graft as an adjunct tool for better decision making. METHODS: Kidney transplanted patients with acute azotemia and documented volume and finally a biopsy were enrolled in this study|. Graft volumes between rejected patients (antibody-mediated rejection {AMR} and cell - mediated rejection {CMR}) and |non rejected but azotemic patients were compared. RESULTS: A total of 76 patients were enrolled in this study |(45 case and 31 control|). 53.3% of the case group were| (AMR)| and 46.7% belonged to |(CMR). There was no difference between kidney volume according to age or sex. But the case group had a significantly bigger volume than controls (253.09 cm3 and 186.45 cm3; p< 0.001). In addition, there was a significant difference between the volumes of AMR kidneys with CMR and controls |(286.24+66.70|, 224.08+76.79 and 186.95+39.92; P=0.003 and p<0.001, respectively), but not between CMR and controls |(P=0.067). A cutoff point of 200 cm3 was determined as rejection with sensitivity and specificity of 70% and a cutoff point of 250 cm3 could be used as AMR cut off with sensitivity of 76% and specificity of 70%. CONCLUSION: There was a significant difference in volume between rejection and control group and between AMR and CMR. So, kidney volume determination is an easy and valuable tool to help the clinician to have a more rapid and better decision making.

Correction to: A downstream box fusion allows stable accumulation of a bacterial cellulase in Chlamydomonas reinhardtii chloroplasts.

[This corrects the article DOI: 10.1186/s13068-018-1127-7.].

Carotenoid Metabolism in Plants: The Role of Plastids.

Carotenoids are indispensable to plants and critical in human diets. Plastids are the organelles for carotenoid biosynthesis and storage in plant cells. They exist in various types, which include proplastids, etioplasts, chloroplasts, amyloplasts, and chromoplasts. These plastids have dramatic differences in their capacity to synthesize and sequester carotenoids. Clearly, plastids play a central role in governing carotenogenic activity, carotenoid stability, and pigment diversity. Understanding of carotenoid metabolism and accumulation in various plastids expands our view on the multifaceted regulation of carotenogenesis and facilitates our efforts toward developing nutrient-enriched food crops. In this review, we provide a comprehensive overview of the impact of various types of plastids on carotenoid biosynthesis and accumulation, and discuss recent advances in our understanding of the regulatory control of carotenogenesis and metabolic engineering of carotenoids in light of plastid types in plants.

A downstream box fusion allows stable accumulation of a bacterial cellulase in Chlamydomonas reinhardtii chloroplasts.

BACKGROUND: We investigated strategies to improve foreign protein accumulation in the chloroplasts of the model algae Chlamydomonas reinhardtii and tested the outcome in both standard culture conditions as well as one pertinent to algal biofuel production. The downstream box (DB) of the TetC or NPTII genes, the first 15 codons following the start codon, was N-terminally fused to the coding region of cel6A, an endoglucanase from Thermobifida fusca. We also employed a chimeric regulatory element, consisting of the 16S rRNA promoter and the atpA 5'UTR, previously reported to enhance protein expression, to regulate the expression of the TetC-cel6A gene. We further investigated the accumulation of TetC-Cel6A under N-deplete growth conditions. RESULTS: Both of the DB fusions improved intracellular accumulation of Cel6A in transplastomic C. reinhardtii strains though the TetC DB was much more effective than the NPTII DB. Furthermore, using the chimeric regulatory element, the TetC-Cel6A protein accumulation displayed a significant increase to 0.3% total soluble protein (TSP), whereas NPTII-Cel6A remained too low to quantify. Comparable levels of TetC- and NPTII-cel6A transcripts were observed, which suggests that factors other than transcript abundance mediate the greater TetC-Cel6A accumulation. The TetC-Cel6A accumulation was stable regardless of the growth stage, and the transplastomic strain growth rate was not altered. When transplastomic cells were suspended in N-deplete medium, cellular levels of TetC-Cel6A increased over time along with TSP, and were greater than those in cells suspended in N-replete medium. CONCLUSIONS: The DB fusion holds great value as a tool to enhance foreign protein accumulation in C. reinhardtii chloroplasts and its influence is related to translation or other post-transcriptional processes. Our results also suggest that transplastomic protein production can be compatible with algal biofuel production strategies. Cells displayed a consistent accumulation of recombinant protein throughout the growth phase and nitrogen starvation, a strategy used to induce lipid production in algae, led to higher cellular heterologous protein content. The latter result is contrary to what might have been expected a priori and is an important result for the development of future algal biofuel systems, which will likely require co-products for economic sustainability.

Artificial Ureter in Patients with Extensive Ureteral Damage.

PURPOSE: loss of significant lengths of ureter when substitution with bowel or bladder fails is a disaster in urology. This study is conducted to evaluate the results of subcutaneous nephron-vesical bypass (SNVB) in ureteral damage of different etiologies. MATERIALS AND METHODS: Seventeen SNVB were employed in patients with ureteral injuries. We employed a device consisted of an internal silicone tube covered by a coiled PTFE tube to replace the ureter. This is called artificial ureter (AU). Proximal end of the AU was introduced in the kidney percutaneously, the tube was passed through a subcutaneous tunnel, while the distal end was inserted in the bladder through a small suprapubic incision. RESULTS: Follow-up ranged from six months to ten years. We removed the prosthetic ureter in one patient due to gross hematuria two months after insertion. One of the patients was reoperated two days after the procedure because of urinary leakage. In all other patients, the procedure was safe and effective. CONCLUSION: Subcutaneous nephron-vesical bypass is a safe and appealing alternative to a nephrostomy tube. This is a permanent device with no need for exchange. The technique can be applied in ureteral injuries due to various causes.