Advancements and prospects of CRISPR/Cas9 technologies for abiotic and biotic stresses in sugar beet.

Sugar beet is a crop with high sucrose content, known for sugar production and recently being considered as an emerging raw material for bioethanol production. This crop is also utilized as cattle feed, mainly when animal green fodder is scarce. Bioethanol and hydrogen gas production from this crop is an essential source of clean energy. Environmental stresses (abiotic/biotic) severely affect the productivity of this crop. Over the past few decades, the molecular mechanisms of biotic and abiotic stress responses in sugar beet have been investigated using next-generation sequencing, gene editing/silencing, and over-expression approaches. This information can be efficiently utilized through CRISPR/Cas 9 technology to mitigate the effects of abiotic and biotic stresses in sugar beet cultivation. This review highlights the potential use of CRISPR/Cas 9 technology for abiotic and biotic stress management in sugar beet. Beet genes known to be involved in response to alkaline, cold, and heavy metal stresses can be precisely modified via CRISPR/Cas 9 technology for enhancing sugar beet's resilience to abiotic stresses with minimal off-target effects. Similarly, CRISPR/Cas 9 technology can help generate insect-resistant sugar beet varieties by targeting susceptibility-related genes, whereas incorporating Cry1Ab and Cry1C genes may provide defense against lepidopteron insects. Overall, CRISPR/Cas 9 technology may help enhance sugar beet's adaptability to challenging environments, ensuring sustainable, high-yield production.

Post-harvest biology and recent advances of storage technologies in sugarcane.

Sugarcane deteriorates at a quick rate, just like other perishable crops. The quick loss of sucrose content in sugarcane from the time it is harvested has a significant impact on sugar recovery. This problem of post-harvest sucrose losses in sugarcane is a serious concern in cane-producing countries, as it not only leads to low sugar recovery in mills, but also to poor sugar refining. Unreasonable delays in cane transportation from the fields to the mill are frequently linked to a number of problems related to primary or secondary sucrose losses, all of which contribute to a significant reduction in cane weight and sugar recovery. In sugar mills, the processing of damaged or stale canes also presents a number of challenges, including increased viscosity due to dextran generation, formation of acetic acid, and dextrans due to Leuconostoc spp. invasion, and so on. The combination of all of these variables results in low sugar quality, resulting in significant losses for sugar mills. The primary and secondary losses caused by post-harvest sucrose degradation in sugarcane are enlisted. The employment of physico-chemical technologies in farmers' fields and sugar mills to control and minimize these losses has also been demonstrated.

Impact of chemical treatments on Leuconostoc bacteria from harvested stored cane/stale cane.

Post-harvest sucrose losses are always a critical problem for sugar industries. A predominant factor which is causing these post-harvest losses that affects sugar recovery is the bacterium Leuconostoc spp. This study aims to check the efficacy of certain chemical treatments in reducing the proliferation of this bacterium. Our study based on a Leuconostoc-specific media revealed that application of 0.5 % aqueous solution of benzalkonium chloride and sodium metasilicate (BKC + SMS), formaldehyde, glutaraldehyde, sodium chloride and pine oil showed significant reduction in zone of proliferation. Considering formaldehyde and glutaraldehyde as control, the most effective treatments were chemical formulations of benzalkonium chloride along with sodium metasilicate, pine oil and sodium chloride in checking the proliferation of this bacterium. The application of these treatments has an immense potential in the sugar industry for reducing post-harvest sugar losses.

Morphological assessment of water stressed sugarcane: A comparison of waterlogged and drought affected crop.

Abiotic stress is recurrent occurring problem for sugarcane crop in terms of hindrance in achieving good and high production. In India, drought coverage is 2.97 lakh ha while 2.5 lakh ha under coverage of waterlogging which is one of the reasons behind low cane production and productivity due to alteration in metabolism, growth and development of the plant either in direct or indirect way. Therefore, we investigated the comparison of morphological losses in drought and waterlogging sugarcanes. Morphological parameters assessed were leaf length, leaf width, leaf area, stalk diameter, cane height, cane weight, internodes number and average internodal length. Also, total root weight, dry matter production of stalk, leaves and roots were observed. Results showed that leaf length was marginally increased in drought canes but it was not so in case of waterlogged canes. Besides, there was decrease in total root weight of sugarcane affected by drought by 16.99% while there was increase by 10.06% in waterlogging affected canes in comparison to normal grown canes. In cane height and stalk diameter, decrease by 18.28%, 7.52%, respectively, in drought and 11.41%, marginal decrease, respectively, in waterlogged affected canes as compared to normally grown canes. Average internodal length was also found to increase in both drought as well as waterlogged canes by 39.02% and 36.60%, respectively, in comparison to normal grown canes. Number of internodes was decreased more in drought affected canes than in waterlogged canes. This study concluded that there are higher morphological losses in sugarcane in drought condition than in waterlogging conditions with respect to normal grown canes.

Minimization of post-harvest sucrose losses in drought affected sugarcane using chemical formulation.

Sugarcane is a significant crop for production of sugar and ethanol in the world. In present perspective, drought is one of the frequently occurring abiotic stresses hampering the productivity of sugarcane causing heavy losses in sugar recovery. Post-harvest sugarcane deterioration attains more importance. Measures have been recommended in harvested canes to prevent these losses in general and under drought conditions but application of chemical formulation has not yet been tested over drought effected ones. Thus, we tried to investigate the efficacy of chemical formulation [Benzalkonium chloride (BKC) + Sodium metasilicate (SMS)] on sucrose losses occurring in harvested canes grown under drought and normal conditions. Results showed that application of chemical formulation had higher effect on drought canes in comparison to normal grown canes. Loss in cane weight was reduced to 8.25% and 11% in drought treated and normal treated grown canes, respectively, after 240 h of harvest in comparison to their respective control. In sucrose content and Commercial cane sugars %, drought treated canes showed an effect of BKC + SMS by reducing the losses to 1.26 units and 1.42 units, respectively, whereas in normal ones, reduction was of 0.38 units and 0.10 units, respectively. Biochemical analysis revealed that in reducing sugars, reduction in increase were of 44.51% and 25.50% in drought and normal grown canes, respectively, after 240 h of harvest. Dextran and soluble acid invertase estimations revealed that after application of BKC + SMS, reduction of dextran and invertase activity were of 49.74%, 66.84%, respectively, and 33.92%, 42.75%, respectively, in drought and normal grown canes, respectively. Total microbial load, showed effectiveness of 25.01% in drought grown canes while 14.41% in normal grown ones after 240 h of harvest. Our study was planned to use the anti-bacterial efficiency of both the chemicals over harvested canes so that the major sucrose losses occurring due to microbial deterioration could be inhibited. The use of this chemical formulation proves to be an effective one over post-harvest sucrose losses, particularly in drought grown canes.

Characterization of rice (Oryza sativa L.) genotypes on the basis of morpho-physiological and biochemical traits grown under aerobic situation in rainfed ecosystem .

The objective of the present study was to examine the effect of aerobic situation on yield, physiological and biochemical traits of advanced breeding lines of rice. Experiment was conducted with two set of rice genotypes under two water regimes (aerobic and irrigated), during three consecutive wet seasons 2010-2012. Significant decrease in yield was observed in rice genotypes grown under aerobic situation as compared to the irrigated ones. Promising rice genotypes having the ability to maintain high plant biomass, harvest index, early vegetative vigour, improved physiological and biochemical traits in terms of relative water content (RWC), leaf area index (LAI), total soluble sugar, starch, protien and proline content help to sustain higher grain yield under aerobic situation. The yield gap between aerobic and irrigated rice ranged between 24% to 68%. Grain yield showed positive correlation with harvest index (0.434), test weight (0.647), plant biomass (0.411) and effective tiller numbers (0.473), whereas spikelet sterility was negative associated (-0.380). The current study suggested that promising genotypes viz., IR77298-14-1-2-130-2, IR84899-B-182-3-1-1-2, IR84887-B-157-38-1-1-3 and IR 84899-B-179-1-1-1-2 for aerobic situation, showing yield advantage due to better performance of physiological and biochemical traits, might be adopted in large area of rainfed ecosystem as well as in irrigated areas where water scarcity was a major problem.

Mismatch repair deficiency in colorectal cancer patients in a low-incidence area.

BACKGROUND: In a previous study we identified 206 patients with colorectal adenocarcinoma in the Northern Cape province of South Africa, diagnosed between January 2002 and February 2009. The age-standardised incidence was 4.2/100 000 per year world standard population. This is 10% of the rate reported in First-World countries. In high-incidence areas, the rate of abnormal mismatch repair gene expression in colorectal cancers is 2 - 7%. OBJECTIVES: The aim of this study was to determine the prevalence of hMLH1- and hMSH2-deficient colorectal cancer in the Northern Cape. METHODS: Formalin-fixed paraffin wax-embedded tissue blocks from 87 colorectal adenocarcinomas identified in the previous study were retrieved. Standard immunohistochemical staining methods were used to detect the expression of hMLH1 and hMSH2 (i.e. products of the hMLH1 and hMSH2 genes) in the tumours using heat-induced antigen retrieval and diaminobenzidene as a chromogen. Results. In 8 blocks there was insufficient tumour tissue and in 1 case the immunohistochemical staining failed, probably owing to poor fixation, leaving 78 cases for analysis. In 11 cases hMLH1 was deficient and in 6 cases hMSH2 was deficient. Overall, 21.8% of cancers were deficient for hMLH1 or hMSH2. CONCLUSION: Presuming that 80% of all hMLH1 deficiencies are due to hypermethylation of the gene, we found 10.5% of colorectal cancers in an area with a low incidence of colorectal cancer to be deficient in the product of the mismatch repair gene/s. This is approximately three times the reported rate in high-incidence areas.

Prevalence and pattern of congenital heart disease in school children of eastern Uttar Pradesh.

BACKGROUND AND AIM: The prevalence of congenital heart disease (CHD) is not known in our country. The aim of present study was to find out the prevalence of CHD in school children of eastern Uttar Pradesh. METHOD: A team consisting of a cardiologist, physicians and junior residents visited schools in the area. All the children were examined for presence of cardiac murmur or history of heart disease or any intervention. Those with murmurs or previous history of heart disease were called to the Medical College Hospital for evaluation by ECG, chest X-ray and echocardiography for confirmation of the lesion. RESULTS: Out of 118,212 children examined, 142 were found to have CHD. The prevalence was 1.3 per 1000 children and the commonest lesions were ventricular and atrial septal defects, aortic stenosis with or without regurgitation, and pulmonary stenosis. CONCLUSION: CHD prevalence is 1.3 per 1000 school children that is nearly two and a half times more than that of RHD. Knowing it is important for development of facilities for CHD care in our setup.

Risk of blood splashes to the eye during surgery.

BACKGROUND: With the advent of a new infectious era involving the HIV and hepatitis B and C viruses, concern has arisen about transmission of these viruses through ocular blood splashes during surgery. The purpose of the study was to determine the risk of ocular blood splashes to surgeons and their assistants during surgery. METHOD: Surgeons and assistants in several surgical disciplines were requested to wear facemasks with a transparent plastic visor. The visors were collected postoperatively and inspected for macroscopic and microscopic blood splashes. RESULTS: Fifty-nine per cent of the surgeons and assistants refused to wear facemasks with a visor. The incidence of blood splashes in those who participated was 45%. There was a trend for blood splashes to be more common during major surgery and during elective surgery. Surgeons and assistants were subject to similar risk. CONCLUSION: This study confirms the significant risk of ocular blood splashes during surgery, while also suggesting that both surgeons and assistants lack appreciation of the risk.

Effect of liver regeneration on the pharmacokinetics of immunosuppressive drugs.

In liver transplantation the graft has been known to undergo regeneration, which is associated with down-regulation of the cytochrome P450 system. The latter is involved in the metabolism of several immunosuppressive drugs. The aim of this study was to investigate the effect of liver regeneration on the pharmacokinetics (PK) of cyclosporine, rapamycin, and tacrolimus. Rats were subjected to either partial hepatectomy (PH) or sham operation (SH). Cyclosporine, rapamycin, and tacrolimus PK studies were performed at 0, 24, and 96 hours postoperatively. The areas under the curve (AUC), trough levels, and maximum concentrations (Cmax) for cyclosporine and tacrolimus were numerically higher in the animals subjected to PH. The PK studies of rapamycin were not affected by PH. These studies indicated that cyclosporine and tacrolimus metabolism may be inhibited during liver regeneration.

Analysis of mucins: role in laboratory diagnosis.

Mucins are high molecular weight glycoproteins with complex oligosaccharide side chains attached to the apomucin protein backbone by O-glycosidic linkage; they are found in crude mucus gels that protect epithelial surfaces in the major tracts of the body and as transmembrane proteins expressed on the apical cell surface of glandular and ductal epithelia of various organs. Changes in the sequence of glycosylation of mucins in different settings generate a variety of epitopes in the oligosaccharide side chains of mucins, including newly expressed blood-group antigens, distinguishing between normal and diseased states. Tumour-associated epitopes on mucins and their antigenicity make them suitable as immunotargets on malignant epithelial cells and their secretions, creating a surge of interest in mucins as diagnostic and prognostic markers for various diseases, and even influencing the design of mucin-based vaccines. This review discusses the emerging roles of mucins such as MUC1 and MUC4 in cancer and some other diseases, and stresses how underglycosylated and truncated mucins are exploited as markers of disease and to monitor widespread metastasis, making them useful in patient management. Furthermore the type, pattern and amount of mucin secreted in some tissues have been considered in the classification and terminology of neoplasia and in specific organs such as the pancreas. These factors have been instrumental in pathological classification, diagnosis and prognostication of neoplasia.

Biochemical and immunohistochemical characterisation of mucins in 8 cases of colonic disease--a pilot study.

OBJECTIVES: To characterise mucins in cancer of the colon and compare these with controls using stringent biochemical measures to avoid endogenous proteolysis. DESIGN: Crude mucus scrapings were collected from 12 specimens obtained by colectomy. Specimens from 3 traumatic colectomies and 1 sigmoid volvulus were used as controls, and compared with 6 specimens from colons resected for adenocarcinoma and 2 irradiated colons. SUBJECTS: The median age of the 4 female patients was 76 years (range 49 - 82 years), and of the 8 male patients 46.5 years (range 16 - 74 years). RESULTS AND CONCLUSIONS: The crude mucus scrapings in the 9 specimens ranged in weight from 353 mg to 7 697 mg (median 4 928 mg). The median of purified mucin in the 9 specimens was 0.72 microg/mg wet weight of scraped material. Eight samples gave non-extractable pellet material, and were treated with DTT to reduce disulphide bonds for further analysis. One of these 8 pellets was resistant to reduction and had to be digested with papain before analysis. Only 5 of these pellets had mucin. Gel filtration and SDS-PAGE (sodium dodecyl sulphate polyacrylamide gel electrophoresis) analysis revealed different populations of mucin based on size and extent of degradation. Western blotting and immunohistochemical analysis confirmed the presence of MUC2 in all samples, MUC5AC in 2 and MUC5B in 5 diseased specimens. Immunohistochemical analysis showed that there was no MUC1 in the normal specimens, MUC1 apoprotein (MUC1 core) in 2 cancer specimens and MUC1 in 1 cancer specimen. Histochemical analysis showed that normal tissue expressed neutral and acidic mucins and diseased specimens predominantly expressed acidic mucins. The electrophoretic behaviour of MUC2 in sigmoid volvulus was different from that in cancer of the colon.

Restoration of liver mass after partial hepatectomy--implications for living donor liver transplantation.

In living donor liver transplantation, the recipient liver undergoes more rapid regeneration than the remnant liver in the donor. In this study we investigated the factors which may be responsible for the difference in the regenerative response between the donor and the recipient. Long Evans rats were subjected to either partial hepatectomy (PH) or sham operation (SH) and were treated with liver cytosol (C) and cyclosporine (Cy). The rats were sacrificed at 24, 48, 72 and 96 hours and 1 and 2 weeks postoperatively. The livers were removed to determine the liver weight/body weight (LW/BW) ratio and the mitotic index. The mitotic index, serum aspartate transferase (AST) and serum alanine transferase (ALT), although unchanged in the SH groups, were increased in the rats treated with PH + C + Cy, and were greater than after PH only. However LW/BW ratios increased after PH but had returned to preoperative levels by 2 weeks. The changes in LW/BW ratio were not modified by the cytosol or cyclosporine.

The effect of rapamycin on the healing of the ureteric anastomosis and wound healing.

Rapamycin is a potent immunosuppressive agent that also inhibits fibroblastic activity and therefore may affect the healing of various tissues. The aim of this study was to investigate the effect of rapamycin on wound healing and the healing of the ureteric anastomosis. Large White/Landrace pigs were subjected to a laparotomy and division and immediate anastomosis of the ureter. The animals were randomly allocated to receive either rapamycin or placebo. The animals were sacrificed on postoperative day 5, and strips of the skin and fascia closure and the ureteric anastomosis excised and used to determine the tensile strength, hydroxyproline levels, and histological changes. The tensile strength and the hydroxyproline levels in the ureter and fascia were lower in the rapamycin-treated animals. There was no difference in the tensile strength in the skin, although the hydroxyproline levels were lower. This study shows that healing of the ureteric anastomosis and fascia and skin closure may be impaired by rapamycin.

Effect of rapamycin on the healing of the bile duct.

Besides its potent immunosuppressive properties, rapamycin also has antitumor and antifungal effects. Rapamycin also inhibits the proliferation of fibroblasts and therefore may impair the healing of various tissues. We investigated the effect of rapamycin on the healing of the bile duct anastomosis. The study was undertaken in pigs that were subjected to a laparotomy under general anesthesia. The bile duct was mobilized and divided and immediately reanastomosed. The animals were randomly allocated to receive either rapamycin or placebo. The animals were sacrificed on the postoperative day 5, then the biliary anastomosis was excised and used to determine the tensile strength, hydroxyproline levels, and the histological changes. The tensile strength and the hydroxyproline levels in the biliary anastomosis were lower in the animals treated with rapamycin. The liver function tests were normal. These studies show that rapamycin may impair the healing of the biliary anastomosis.

Restoration of liver mass after partial hepatectomy - implications for living donor liver transplantation

Abstract In living donor liver transplantation; the recipient liver undergoes more rapid regeneration than the remnant liver in the donor. In this study we investigated the factors which may be responsible for the difference in the regenerative response between the donor and the recipient. Long Evans rats were subjected to either partial hepatectomy (PH) or sham operation (SH) and were treated with liver cytosol (C) and cyclosporine (Cy). The rats were sacrificed at 24; 48; 72 and 96 hours and 1 and 2 weeks postoperatively. The livers were removed to determine the liver weight/body weight (LW / BW) ratio and the mitotic index. The mitotic index; serum aspartate transferase (AST) and serum alanine transferase (ALT); although unchanged in the SH groups; were increased in the rats treated with PH + C + Cy; and were greater than after PH only. However LW / BW ratios increased after PH but had returned to preoperative levels by 2 weeks. The changes in LW / BW ratio were not modified by the cytosol or cyclosporine

Hypoalbuminaemia in brain-dead donors for liver transplantation: transplantation

Liver transplantation has become established as the treatment of choice for most patients with end-stage liver disease and is performed on a routine basis in most major centres throughout the world. The majority of donors for liver transplantation are brain-dead cadaver donors following either a severe head injury or a massive intracranial haemorrhage. Potential liver donors undergo a rigid screening process before being accepted. This includes a thorough clinical examination to assess the haemodynamic status of the donor and to exclude any overt evidence of liver disease. Blood samples are also taken for viral studies to exclude HIV infection and hepatitis B and C infection; and for liver function tests to exclude liver disease or liver injury. Over the years we have noted that our liver donors often had low serum albumin levels; although this has not been formally documented. A review of the literature revealed that hypoalbuminaemia associated with severe head injury has been documented previously. However the impact of brain death on serum albumin levels has not been studied previously. The present study was therefore undertaken to document serum albumin levels in brain-dead cadaver donors

Restoration of liver mass after partial hepatectomy - implications for living donor liver transplantation : transplantation

In living donor liver transplantation; the recipient liver undergoes more rapid regeneration than the remnant liver in the donor. In this study we investigated the factors which may be responsible for the difference in the regenerative response between the donor and the recipient. Long Evans rats were subjected to either partial hepatectomy (PH) or sham operation (SH) and were treated with liver cytosol (C) and cyclosporine (Cy). The rats were sacrificed at 24; 48; 72 and 96 hours and 1 and 2 weeks postoperatively. The livers were removed to determine the liver weight/body weight (LW / BW ) ratio and the mitotic index. The mitotic index; serum aspartate transferase (AST) and serum alanine transferase (ALT); although unchanged in the SH groups; were increased in the rats treated with PH + C + Cy; and were greater than after PH only. However LW / BW ratios increased after PH but had returned to preoperative levels by 2 weeks. The changes in LW / BW ratio were not modified by the cytosol or cyclosporine