Microbial production of N-acetyl-D-glucosamine (GlcNAc) for versatile applications: Biotechnological strategies for green process development.

N-acetyl-d-glucosamine (GlcNAc) is a commercially important amino sugar for its wide range of applications in pharmaceutical, food, cosmetics and biofuel industries. In nature, GlcNAc is polymerised into chitin biopolymer, which is one of the major constituents of fungal cell wall and outer shells of crustaceans. Sea food processing industries generate a large volume of chitin as biopolymeric waste. Because of its high abundance, chitinaceous shellfish wastes have been exploited as one of the major precursor substrates of GlcNAc production, both in chemical and enzymatic means. Nevertheless, the current process of GlcNAc extraction from shellfish wastes generates poor turnover and attracts environmental hazards. Moreover, GlcNAc isolated from shellfish could not be prescribed to certain groups of people because of the allergic nature of shell components. Therefore, an alternative route of GlcNAc production is advocated. With the advancement of metabolic construction and synthetic biology, microbial synthesis of GlcNAc is gaining much attention nowadays. Several new and cutting-edge technologies like substrate co-utilization strategy, promoter engineering, and CRISPR interference system were proposed in this fascinating area. The study would put forward the potential application of microbial engineering in the production of important pharmaceuticals. Very recently, autotrophic fermentation of GlcNAc synthesis has been proposed. The metabolic engineering approaches would offer great promise to mitigate the issues of low yield and high production cost, which are major challenges in microbial bio-processes industries. Further process optimization, optimising metabolic flux, and efficient recovery of GlcNAc from culture broth, should be investigated in order to achieve a high product titer. The current study presents a comprehensive review on microbe-based eco-friendly green methods that would pave the way towards the development of future research directions in this field for the designing of a cost-effective fermentation process on an industrial setup.

N-Acetyl-D-glucosamine Production by a Chitinase of Marine Fungal Origin: a Case Study of Potential Industrial Significance for Valorization of Waste Chitins.

Chitin is a linear homo-polymer of N-acetyl-D-glucosamine (GlcNAc) and the second most abundant biopolymer after cellulose. Several industries rely on the bioprocesses for waste chitin recycle and hydrolysis by chitinase (EC 3.2.1.14) for potential healthcare applications through the production of its monomeric subunit, GlcNAc. In the present study, a chitinase-producing fungus (named as MFSRK-S42) was isolated from the marine water sample of North Bay of the Andaman and Nicobar Islands. It was identified as Aspergillus terreus by morphological and molecular characterization methods leveraging the internal transcribed spacer between 18S rRNA and 5.8S rRNA. Chitinase that was isolated from the fermentation broth of marine Aspergillus terreus was used to carry out biotransformation of chitineaceous wastes. Prior to the enzymatic hydrolysis step, chitins from different sources were characterized for the presence of characteristic functional groups, grain size distribution, and surface morphology. Enzymatic hydrolysis of 50 mg/ml substrate with six units of enzyme incubated for 5 days revealed 15, 36.5, 40, and 46 mg/ml GlcNAc production from ground prawn shell, chitin flakes, colloidal prawn shell, and swollen chitin respectively under standardized conditions, as determined by HPLC. In this study, 30, 73, 80, and 92% GlcNAc yields were observed from ground prawn shell, chitin flakes, colloidal prawn shell, and swollen chitin conversion respectively. The HPLC-eluted product was confirmed as GlcNAc by the presence of characteristic functional groups in FTIR and 244 Da molecular weight peak in HRMS analyses.

Fixed bed column study for water defluoridation using neem oil-phenolic resin treated plant bio-sorbent.

Fluoride has both detrimental and beneficial effects on living beings depending on the concentration and consumption periods. The study presented in this article investigated the feasibility of using neem oil phenolic resin treated lignocellulosic bio-sorbents for fluoride removal from water through fixed bed column study. Results indicated that treated bio-sorbents could remove fluoride both from synthetic and groundwater with variable bed depth, flow rate, fluoride concentration and column diameter. Data obtained from this study indicated that columns with the thickest bed, lowest flow rate, and fluoride concentration showed best column performance. Bio-sorbents used in this study are regenerable and reusable for more than five cycles. The initial materials cost needed to remove one gram of fluoride also found to be lower than the available alternatives. This makes the process more promising candidate to be used for fluoride removal. In addition, the process is also technically advantageous over the available alternatives.

In situ synthesis and antibacterial activity of copper nanoparticle loaded natural montmorillonite clay based on contact inhibition and ion release.

Copper nanoparticle based clay composite has been synthesized by in situ reduction of a copper ammonium complex ion and characterized by different analytical instruments. The copper nanoparticles were both intercalated and adsorbed on the surface with diameters of <5nm (for intercalated) and 25-30nm (for adsorbed). The composite showed good stability for over 3 months in air. Excellent antimicrobial activity of the composite was observed on Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Enterococcus faecalis with mortality rates >90% after 12h. Cellular membrane damage permeated by direct attachment of the composite and indirect damage caused by released copper ion are the primary sources of antibacterial action. Cytotoxicity measurements showed minimal adverse effect on the two human cell lines beyond the M.B.C. value for the microorganisms studied. In the present form the clay composite shows good promise for use in therapeutic applications.

Enhanced biodegradation resistance of biomodified jute fibers.

A bio-catalyzed process has been developed for treating jute fibers to enhance their tensile strength and resistance against biodegradation. Lipolytic bacteria were used in the process to transesterify jute fibers by replacing hydrophilic hydroxyl groups within cellulose chains with hydrophobic fatty acyl chains. Transesterification of some of the hydroxyl groups within the fiber was confirmed with FTIR, UV-vis spectroscopy, (13)C solid state NMR, gas chromatography and analytical determination of ester content. Biomodified fibers exhibited remarkably smaller affinity to water and moisture and retained 62% of their initial tensile strengths after being exposed to a composting environment over 21 days. The corresponding figure for untreated fibers was only 30%. Efficacy of the process reported herein in terms of tensile strength and biodegradation resistance enhancement of fibers achieved after treatment appears to be comparable with similar chemical processes and better than the enzyme-catalyzed alternatives.

A prospective randomised, controlled clinical trial comparing medial and lateral entry pinning with lateral entry pinning for percutaneous fixation of displaced extension type supracondylar fractures of the humerus in children.

OBJECTIVE: To compare the efficacy of medial and lateral entry pinning with lateral entry pinning for percutaneous fixation of displaced (Gartland type II and type III) extension type supracondylar fractures of the humerus in children. METHODS: The study was a single center, prospective, randomized controlled clinical trial. Between October 2007 and September 2010, 160 patients who satisfy the inclusion and exclusion criterias were enrolled in the study, with 80 patients in each group. All the percutaneous pinning was done according to a uniform standardized technique. The patients were re-evaluated as outpatients at three weeks, six weeks and three months after the surgery. At three months follow-up visit, following informations were recorded as outcome measures: (i) Carrying angle (deg) (ii) passive range of elbow motion (deg) (iii) Flynn's criteria for grading, based on the loss of carrying angle and loss of total range of elbow motion. (iv) Baumann angle (deg) (v) Change in Baumann angle (deg) between the Intraoperative radiographs after the surgery and radiographs at three months follow-up visit (vi) loss of reduction grading, based on the change in the Baumann angle. RESULTS: There were no significant differences between the two groups with regard to base-line characteristics, withdrawals and complication rate. At three months follow-up visit, patients were evaluated by recording the various outcome measures. There were no significant differences between the two groups with regard to the various outcome measures such as carrying angle, passive range of elbow motion, Flynn grading, Baumann angle, change in the Baumann angle and loss of reduction grading. CONCLUSIONS: If a uniform standardized operative technique is followed in each method, then the result of both the percutaneous fixation methods will be same in terms of safety and efficacy.

Measles in rural West Bengal, India, 2005-6: low recourse to the public sector limits the use of vitamin A and the sensitivity of surveillance.

BACKGROUND: Measles is often underreported. We evaluated the sensitivity of the measles surveillance in 2 districts of West Bengal in 2005-2006. METHODS: We sampled households with children aged <5 years in village clusters selected with probability proportional to size. We searched households door to door to identify World Health Organization-defined suspected measles cases that had occurred during 12 months in 2004-2005 in Howrah and in 2006 in Purulia. We interviewed mothers about use of health care services during episodes and calculated the proportion of patients seen in the public sector. We reviewed surveillance records at all levels to estimate the proportion of cases seen in public health care facilities that had been reported to the district. We calculated the overall sensitivity of measles surveillance by multiplying these 2 proportions. RESULTS: In Howrah, we identified 240 cases of measles. Of these, 8 (3.3% [95% confidence interval {CI}, 1.5%-6.5%]) had been seen in public facilities and recorded. Of 980 cases identified in 448 public facilities in the periphery, 962 (98%) had been transmitted to the district (overall sensitivity of surveillance, 3.2%). In Purulia, we identified 167 measles cases. Of these, 39 (23.4% [95% CI, = 17.2%-30.5%]) had been seen in public facilities and recorded. Of 418 cases identified in public facilities in the periphery, 414 (99%) had been transmitted to the district (overall sensitivity of surveillance, 23.1%). CONCLUSIONS: Measles surveillance captured a minority of measles cases, but cases captured were transmitted well to the district. Surveillance must engage the private sector. Health education focusing on vitamin A treatment for measles might provide an incentive to seek care, which could increase the sensitivity of surveillance.

Efficacy of chloroquine and sulphadoxine-pyrimethamine either alone or in combination before introduction of ACT as first-line therapy in uncomplicated Plasmodium falciparum malaria in Jalpaiguri District, West Bengal, India.

OBJECTIVE: In India, till recently, Chloroquine was used as first-line therapy in areas with Chloroquine sensitive Plasmodium falciparum malaria cases. The National Vector Borne Disease Control Programme (NVBDCP) has introduced artemisinin combination therapy (ACT) as first-line option to treat all P. falciparum cases in the country. This study was carried out to ascertain the efficacy of Chloroquine and Sulphadoxine-Pyrimethamine, either alone or in combination, before the launch of ACT by NVBDCP. METHODS: A total of 300 P. falciparum malaria cases were enrolled randomly in three study arms, Chloroquine (CQ), Sulphadoxine-Pyrimethamine (SP) and Chloroquine plus Sulphadoxine-Pyrimethamine (CQ + SP). All patients were followed up for 28 days as per WHO (Assessment and Monitoring of Antimalarial Drug Efficacy for the Treatment of Uncomplicated Falciparum Malaria, Geneva, 2003) Protocol. Paired blood samples of treatment failure cases were collected and subjected to MSP 1, MSP 2 and GLURP genotyping for differentiation between re-infection and recrudescence. The data were analysed by Kaplan-Meier survival curve according to WHO standard procedures. RESULTS: The overall failure rate including both early treatment failure (ETF) and late treatment failure (LTF) of CQ, SP and CQ + SP were 61%, 14% and 8%, respectively, in the study area. Of 60 recurrent malaria cases, genotyping was successful in 49 cases, revealing that most of the (46/49; 94%) cases of recurrent malaria were due to recrudescence. CONCLUSION: In Jalpaiguri District the overall failure rate of CQ was 61% and of SP 14%, which was well above the WHO recommended cut-off threshold level (10%) for change of drug policy.

Enhancement of tensile strength of lignocellulosic jute fibers by alkali-steam treatment.

The physico-chemical properties of jute fibers treated with alkali (NaOH) solution have been investigated in this study. The treatments were applied under ambient and elevated temperatures and high pressure steaming conditions. To the knowledge of these authors the influence of alkali-steam treatment on the uniaxial tensile strength of natural ligno-cellulosic fibers, such as jute, has not been investigated earlier. The results from this investigation indicate that a 30 min dipping of the fibers in 0.5% alkali solution followed by 30 min alkali-steam treatment leads to an increase in the tensile strength of up to 65%. The increase appears to be due to fiber separation and removal of non-cellulosic materials, which, in turn, resulted in an increased crystallinity.

Prophylactic granisetron vs pethidine for the prevention of postoperative shivering: a randomized control trial.

SUMMARY: Shivering-the "Big Little Problem" has an incidence of 60% in early recovery phase following general anaesthesia. A number of techniques have been tried to prevent postoperative shivering. Previous study showed that, ondansetron in higher doses reduces postoperative shivering. Therefore, this study was done to compare the efficacy of prophylactic granisetron, pethidine and placebo in preventing postoperative shivering. Ninety patients aged 20-60yrs, ASA physical status I and II, scheduled for laparoscopic surgery under general anaesthesia were randomly allocated to receive either normal saline (Group S, n=30) as negative control, pethidine 25mg (Group P, n=30) as positive control or granisetron 40mcg.kg(-1) (Group G, n=30) intravenously before induction. The anaesthesia was induced with fentanyl 2mcg.kg(-1), propofol 2mg.kg(-1) and atracurium 0.5mg.kg(-1) and maintained with sevoflurane 1 - 1.5%. Nasopharyngeal temperature was measured throughout the procedure. An investigator, blinded to the treatment group, graded postoperative shivering in a scale of 0 to 4. (0= no shivering, 1= piloerection or peripheral vasoconstriction but no visible shivering, 2= muscle activity in only one muscle group 3= muscle activity in more than one muscle group, 4= shivering involving the whole body). Prophylaxis was regarded as ineffective if shivering was greater than grade 3 and intravenous pethidine 25 mg was administered as rescue medication. The three groups did not differ significantly regarding patient characteristics. The numbers of patients shivering on arrival in the recovery room at 15 minutes after operation were significantly less in Group P (7%) and Group G (17%) than in Group S (60%). Groups P and G differ significantly than in Group S (p<0.05). However, the difference between Groups P and G was not statistically significant (p>0.05). The prophylactic use of granisetron (40mcg.kg(-1)) and pethidine(25mg) intravenous were found to be effective in preventing postoperative shivering.