Additional Passage Research Articles

Observation and mitigation of perimeter artefacts in bench scale membrane characterization

Perimeter artefacts, such as test cell induced membrane damage and unrepresentative hydrodynamics, can significantly affect bench scale performance evaluation using membrane coupons. By comparing separately collected center and perimeter permeate, the effect of perimeter artefacts can be identified. The additional salt passage in the perimeter area (i.e., along the edges of the membrane coupon) depends non-linearly on pressure, and is implicitly affected by flux, temperature and salinity (osmotic pressure), leading to a potential increase of the measured salt passage by a factor two. We observed a very slow dynamic response (τ≈12h) to changes in operational conditions, suggesting that in our case the perimeter artefact was diffusive in nature. Due to this slow response, the perimeter salt passage trails previously evaluated operational conditions, possibly resulting in a false trend, hysteresis or high variance, depending on the sequential ordering and duration of test condition intervals. The effect of perimeter artefacts can be largely mitigated by measuring salt rejection from only the central area of the coupon.

Poster 114: Viability and Expression of Chondrocytes Harvested During Arthroscopic Treatment of Femoroacetabular Impingement

Poster 114: Viability and Expression of Chondrocytes Harvested During Arthroscopic Treatment of Femoroacetabular Impingement

COMPARATIVE EXPERIMENTAL RESEARCH ON THE QUALITY OF THE WORK OF THE COMBINED EQUIPMENT FOR SHREDDING AND INCORPORATING VEGETABLE RESIDUES COMPARED TO THE AGGREGATE SHREDDER OF VEGETABLE RESIDUES WITH THE U650 TRACTOR

In practice, the operation of destroying plant residues is carried out separately, in general the equipment is mounted on the back of the tractor. Equipment for destroying plant residues such as cutters, platforms with rotors, shredders, etc. is known. which works in aggregate with the tractor. The disadvantages of these equipments consist in the fact that they work independently and only carry out the operation of destroying plant remains and leaving them on the ground, in the technology being the additional passage on the same land for incorporation, which involves fuel consumption, pollution. ground subsidence etc. The combined equipment for shredding and incorporating plant residues eliminates these disadvantages, mainly because the operations are performed in a single pass. The advantage of using this mechanism consists in facilitating operations and increasing productivity with reduced time and fuel costs, at the same time meeting environmental protection requirements by reducing land subsidence and noxes produced by double tillage. Another advantage consists in avoiding the accumulation of plant debris in front of the active organs of the plow and removing it from the furrow, in case the unshredded material would remain on the ground.

In vitro propagation from seeds and enhanced synthesis of podophyllotoxin from root callus of SinoPodophyllum hexandrum Royle T.S. Ying (Himalayan Mayapple) − An endangered medicinal plant

In vitro propagation and secondary metabolite production are the vital biotechnological approaches for the multiplication, conservation, and phytochemicals production from rare, endangered, and industrially important medicinal plants . Himalayan Mayapple ( SinoPodophyllum hexandrum Royle T.S. Ying) is a perennial venerate herb of the alpine Himalayas, rhizomes of which yield podophyllotoxin (PTOX), widely used in the pharmaceutical industry for the synthesis of anti-cancer drugs. The low regeneration rate and high demand for this plant emphasized the need for the development of a non-conventional method for its propagation and PTOX production using biotechnological interventions. In the present study, a single-step protocol for in vitro propagation of S. hexandrum through seeds and PTOX production from seedlings-induced leaf and root calli of three geographical locations in Himachal Pradesh (India) is reported. Maximum in vitro seed germination (56.23%) was obtained after 77 days followed by the highest shoot multiplication on MS medium containing BA (2.0 mg/L), NAA (0.2 mg/L), and calcium chloride (0.7 mg/L) after the fourth sub-culture. Rooting was induced at the base of shoots during in vitro shoot multiplication hence, the shoots were directly transferred for hardening without adopting additional rooting passage. For PTOX production, callus was induced from in vitro leaves and roots on B5 medium supplemented with BA (0.1 mg/L) + NAA (1.0 mg/L) and BA (0.5 mg/L) + 2,4-D (0.5 mg/L) under dark incubation. High-performance liquid chromatography (HPLC) analysis revealed maximum PTOX content (0.27 mg/g) from in vitro root callus of district Kinnaur followed by Lahaul-Spiti (0.26 mg/g), and Kullu (0.25 mg/g), thus signifying the role of high altitudes in enhanced PTOX accumulation. The current study offers an insight into the high-frequency regeneration and PTOX production of S. hexandrum , which may serve as significant baseline information for the production of high-quality plant material in the pharmaceutical industry. • Developed single-step in vitro propagation protocol from seeds of Himalayan Mayapple. • Effect of altitude on enhanced podophyllotoxin production was studied.

High Flux and Antifouling Nanofiltration Membrane Modified by Ag@UiO-66-NH2 and Its Application for Biphenol A Removal

Owing to the specific porous structure which could provide additional passage channel for some molecules, metal organic frameworks are attractive candidates for enhancing permeability and selectivity of membranes in pervaporation, reverse osmosis, and gas separation. In this experiment, Ag@UiO-66-NH2 was introduced into polyamide separation layer by interfacial polymerization of triethylenetetramine and 1,3,5-benzenetricarboxylic acid chloride for nanofiltration. The results indicated that Ag@UiO-66-NH2 nanoparticles did endow the membranes with rapid diffusion pathways for water molecules. When the content of Ag@UiO-66-NH2 was 0.03 g, the prepared membrane (NF-Ag-3) showed high flux about 47.3 L·m-2·h-1 at 0.6 MPa, which is about 2-fold higher than that of polyamide membrane without Ag@UiO-66-NH2, while the MgSO4 rejection rate remained about 87.4%. The membrane also showed excellent antifouling properties, and the water flux recovery ratio was 95.6% after filtration BSA solution. When it was applied for 50 mg/L bisphenol A removal, the rejection rate reached 94.6%, and the flux is about 49.1 L·m-2·h-1. Moreover, Ag particles on UiO-66-NH2 rendered the membrane with good inhibition for Escherichia coli. The antibacterial rate of the membranes is above 95% when the loading of Ag@UiO-66-NH2 is more than 0.03 g.

Experimental Study of Heat Transfer in Concentric Triple Pipe Heat Exchanger

Heat exchangers are devices which are used to transfer the thermal energy between two or more fluids. They are used in various industries for space heating, air conditioning and chemical processing. Some heat exchangers which are experienced in our daily lives include pasteurization, radiators and oil coolers in automobiles Heat exchangers have been divided in several ways, based on its flow direction which are parallel or counter flow, types of construction of the heat exchanger like tubular or plate heat exchanger, the ways of contact between the fluids i.e. direct or indirect.The heat exchangers performance is usually depending upon the physical characteristics of the fluid and the material. When a concentric tube is added in the intermediate space of a double pipe heat exchanger, we obtain a triple pipe heat exchanger, we obtain a triple pipe heat exchanger. The performance of the triple is better than the double pipe, as it provides better heat transfer efficiencies. Basically, the additional pipe improves the heat transfer by providing an additional flow passage and a larger area for heat transfer area per unit length of the exchanger.

Comparative Bioactivity Analysis for Off-the-Shelf and Culture–Rescued Umbilical Cord-Derived Mesenchymal Stem/Stromal Cells in a Xeno- and Serum-Free Culture System

We recently reported a standardized xeno- and serum-free culture platform to isolate and expand umbilical cord-derived mesenchymal stem/stromal cells (UC-MSCs). Comparing populations from the same passage, cells that were cryopreserved and culture-rescued exhibited characteristics similar to those of their fresh counterparts, continuously cultured cells without interim cryopreservation. The culture rescue after thawing allowed for the cells to be fully recovered. However, since it would be more cost-effective and timesaving if cryopreserved cells can be used as an off-the-shelf product, we set out to compare the bioactivity of freshly thawed UC-MSCs versus culture-rescued UC-MSCs of the same batch that were recultured for an additional passage under our xeno- and serum-free protocol. UC-MSCs showed high viability in both the freshly thawed and the re-cultured group. Both populations displayed a similar proliferation capacity which is indicated by a comparable population doubling time and colony-forming ability. Both freshly thawed and culture-rescued UC-MSCs expressed the characteristic immunophenotype and were capable of differentiating into osteocytes, chondrocytes, and adipocytes. On the other hand, culture-rescued cells appeared to be more potent in immunosuppression than freshly thawed cells. In conclusion, freshly thawed and culture-rescued cell products share comparable bioactivity in cell growth and proliferation, immunophenotype, and differentiation potential. However, the culture-rescued cells that were allowed to grow for an additional passage appear to display a more favorable immunomodulatory potential when compared to their freshly thawed parent cells.

IP-10 and CXCR3 signaling inhibit Zika virus replication in human prostate cells.

Our previous studies have shown that Zika virus (ZIKV) replicates in human prostate cells, suggesting that the prostate may serve as a long-term reservoir for virus transmission. Here, we demonstrated that the innate immune responses generated to three distinct ZIKV strains (all isolated from human serum) were significantly different and dependent on their passage history (in mosquito, monkey, or human cells). In addition, some of these phenotypic differences were reduced by a single additional cell culture passage, suggesting that viruses that have been passaged more than 3 times from the patient sample will no longer reflect natural phenotypes. Two of the ZIKV strains analyzed induced high levels of the IP-10 chemokine and IFNγ in human prostate epithelial and stromal mesenchymal stem cells. To further understand the importance of these innate responses on ZIKV replication, we measured the effects of IP-10 and its downstream receptor, CXCR3, on RNA and virus production in prostate cells. Treatment with IP-10, CXCR3 agonist, or CXCR3 antagonist significantly altered ZIKV viral gene expression, depending on their passage in cells of relevant hosts (mosquito or human). We detected differences in gene expression of two primary CXCR3 isoforms (CXCR3-A and CXCR3-B) on the two cell types, possibly explaining differences in viral output. Lastly, we examined the effects of IP-10, agonist, or antagonist on cell death and proliferation under physiologically relevant infection rates, and detected no significant differences. Although we did not measure protein expression directly, our results indicate that CXCR3 signaling may be a target for therapeutics, to ultimately stop sexual transmission of this virus.

Spheroid-Like Cultures for Expanding Angiopoietin Receptor-1 (aka. Tie2) Positive Cells from the Human Intervertebral Disc.

Lower back pain is a leading cause of disability worldwide. The recovery of nucleus pulposus (NP) progenitor cells (NPPCs) from the intervertebral disc (IVD) holds high promise for future cell therapy. NPPCs are positive for the angiopoietin-1 receptor (Tie2) and possess stemness capacity. However, the limited Tie2+ NPC yield has been a challenge for their use in cell-based therapy for regenerative medicine. In this study, we attempted to expand NPPCs from the whole NP cell population by spheroid-formation assay. Flow cytometry was used to quantify the percentage of NPPCs with Tie2-antibody in human primary NP cells (NPCs). Cell proliferation was assessed using the population doublings level (PDL) measurement. Synthesis and presence of extracellular matrix (ECM) from NPC spheroids were confirmed by quantitative Polymerase Chain Reaction (qPCR), immunostaining, and microscopy. Compared with monolayer, the spheroid-formation assay enriched the percentage of Tie2+ in NPCs’ population from ~10% to ~36%. Moreover, the spheroid-formation assay also inhibited the proliferation of the Tie2- NPCs with nearly no PDL. After one additional passage (P) using the spheroid-formation assay, NPC spheroids presented a Tie2+ percentage even further by ~10% in the NPC population. Our study concludes that the use of a spheroid culture system could be successfully applied to the culture and expansion of tissue-specific progenitors.

Selective fish passage: Restoring habitat connectivity without facilitating the spread of a non-native species

River managers are challenged to address two key threats to freshwater biodiversity. The first is the effects of habitat fragmentation by instream structures, such as dams and weirs, that disrupt migrations and impact species distributions. The second is the impact of non-native species on native species and ecological processes. However, mitigating anthropogenic habitat fragmentation through the installation of passage facilities can facilitate the invasion and spread of non-native species. This study compared the potential of two existing low-cost fish passage technologies designed for sloping weirs, a cylindrical bristle cluster (CBC) array and horizontally oriented studded tiles, to facilitate upstream movement of native European fish while preventing dispersal of non-native American signal crayfish (Pacifastacus leniusculus); thus providing a selective fish passage solution. Crayfish movement and passage was experimentally quantified at a Crump weir installed in a recirculating flume under two velocity regimes (low and high), without (control) and with the addition of either a CBC array or studded tiles. Results were compared to passage efficacy (PE) data for native fish species for both technologies (existing data). Most (84.4%) crayfish were active during the trials, exhibiting frequent up and downstream movements below the weir. During control conditions under the high velocity regime, high velocities (ca. 2.39 m s−1) prevented crayfish reaching the foot of the weir (PE: 0%). Under the low velocity regime, relatively low velocities (ca. 0.74 m s−1) at the weir crest prevented most crayfish from passing (PE: 10–16%). Crayfish movement speed and total distance moved were lower under the high than the low velocity regime. Neither fish pass technology improved crayfish maximum distance of ascent on the downstream weir face or PE under either velocity regime. Under comparable conditions to the high velocity regime tested here, previous studies have shown both technologies improve PE for native fish. Hence, both CBC arrays and studded tiles would likely function as suitable selective fish passes where the conservation objective is not to aid the spread of non-native crayfish. Additional passage inhibiting technologies will be required at sites where complete blockage of crayfish movement is required.

In vivo evolution of viral virulence: switching of deformed wing virus between hosts results in virulence changes and sequence shifts.

The health of the Western honey bee is threatened by a global epidemic of deformed wing virus (DWV) infections driven by the ectoparasitic mite Varroa destructor acting as mechanical and biological virus vector. Three different variants of DWV, DWV-A, -B and -C exist. Virulence differences between these variants and their relation to V. destructor are still controversially discussed. We performed laboratory experiments to analyze the virulence of DWV directly isolated from crippled bees (DWVP0 ) or after one additional passage in bee pupae (DWVP1 ). We demonstrated that DWVP0 was more virulent than DWVP1 for pupae, when pupal mortality was taken as virulence marker, and for adult bees, when neurotropism and cognitive impairment were taken as virulence markers. Phylogenetic analysis supported that DWV exists as quasispecies and showed that DWVP0 clustered with DWV-B and DWVP1 with DWV-A when the phylogeny was based on the master sequences of the RNA-dependent RNA polymerase but not so when it was based on the VP3 region master sequences. We propose that switching of DWV between the bee and the mite host is accompanied by changes in viral sequence, tissue tropism and virulence and that the RNA-dependent RNA polymerase is involved in determining host range and virulence.

Investigating the Dehumidification Characteristics of Turbine Stator Cascades with Parallel Channels

Steam turbines play a vital role in the power industry. The efficiency of the steam turbine can be improved by reforming the flow structure. For the White cascade, a parallel channel which is an additional steam passage was designed to connect adjacent steam flow channels. In view of the steepening and sensitive characteristics of the steam condensation parameters in the White cascade, the two-fluid model was applied to study the condensation and aerodynamic characteristics of the prototype cascade and the modified cascade. Wetness losses were regarded as the evaluation standard, and the optimum design of the parallel channel was obtained. The influence of channel width was also studied. The location of the suction surface (D point) has great influence on the dehumidification performance of the cascade. On the contrary, the location of the pressure surface (C point) has no significant effect on dehumidification performance. Moreover, it is necessary to select the width of the parallel channel in combination with the cascade performance. It is suggested that the parallel channel width should be between 3 and 5 mm. The research results can provide a reference for the optimal design of a turbine stator cascade.

Nonlinear modeling and analysis of a shock absorber with a bypass

The model of a mono-tube shock absorber with a bypass is proposed in this paper. It is shown that the application of an additional flow passage (bypass) causes changes to the damping force characteristics when the excitation amplitudes are large. In such cases, the damping force values increase, thereby improving safety of the ride. For small excitation amplitudes, the shock absorber behaves in a similar fashion as shock absorbers without a bypass, ensuring a high comfort level of the ride on roads with smooth surfaces.

THE IMPLEMENTATION OF ACCELERATED TRAINS OF INTERREGION CONNECTION

One of the main directions of improving the efficiency of passenger transportation is the introduction of a system of accelerated trains of inter-regional traffic, subject to the abolition of stops at separate points, which give a slight accumulation of passenger traffic. The paper proposes approaches to reducing the time spent by passengers on the road when traveling in interregional traffic. Reducing the passage time of a passenger train can be provided in two ways: as a result of organizational and technical measures, such as the temporary cancellation of stops at separate points, which give a slight accumulation of passenger traffic and as a result of modernization of permanent devices, which, if necessary, includes and rebuild. The reasons limiting the speed of movement at the railway station are revealed and analyzed. As a result of the cancellation of stops at separate points, the operating costs associated with the additional mechanical work of rolling stock in the process of acceleration, deceleration and parking will be reduced; expenses connected with delay of passengers on the way in the train; but on the other hand, the costs of additional waiting and passage of passengers residing at the stopping point will be increased. To solve the problem, a mathematical model for the scheme of stopping of accelerated trains of interregional communication was developed. To solve, that is, to identify possible stops in a small number of permissible solutions, it is possible to use a method of perforation with a constraint that provides a rigorous approach. The application of this method is based on the following: the criterion, in which the overall effect is taken, weakens the property of addiction, as well as a relatively small number of possible solutions (given the fact that a certain number of stops persists, as well as the fact that achieving a given time reduction the course of canceling one stop is not provided, and at least four or five, the number of possible solutions is further reduced). Stopping points at which stops are canceled are selected based on the maximum effect due to cancellation of the stop, as well as the possibility of modernization of permanent stopping devices.

Effect of geometric modification on flow behaviour and performance of reverse flow combustor

Numerical investigation had been performed on the reverse flow combustor of a mini gas turbine engine so as to investigate the performance characteristics of the combustor by means of geometry modifications. In order to enhance the thrust performance of the reverse flow combustor, the baseline combustor (Model A) was previously modified by increasing its chamber volume by 15%, the fuel-air ratio (FAR) by 40% and by raising the injection point density to two (Model B). However, the thrust optimization of the baseline combustor resulted in high combustor exit temperature that could potentially damage the combustor liners. To rectify the adversity of high exit temperature, the combustor cooling effects were achieved by subsequently adding additional passage holes at the dilution zone of the Model B combustor so as to direct the incoming cold flow from the compressor exit towards the outgoing hot flow in the reverse flow combustor (Model C). The commercial software ANSYS Fluent 17.0 was adopted in this study and to solve the turbulence model, Reynolds Averaged Navier–Stokes methodology was adopted by employing standard k-ɛ turbulence model with standard wall function. A probability density function model was generated to introduce the combustion species and a discrete phase Model was employed to specify the kerosene fuel-based injector properties. The numerical results of Model A combustor were validated against the previous experimental results using grid convergence test. The numerical results were observed to be in good agreement with the experimental results. However, ineffective mixing was found to be a setback for the baseline combustor (Model A) indicating the need for combustor performance improvement. The comparative results of the revised combustor model (Model C) showed that better cooling effects at the combustor exit could be achieved by adding supplementary passage holes at the downstream of the combustor outer liner, respectively. The addition of dilution holes also resolved the issue of high pressure loss that was observed in Model A combustor with no significant change in the specific fuel consumption. The present paper confirms that the performance of the reverse flow combustor model could be affected by slight geometric modification. The performance characteristics of the combustor models are presented in terms of thrust, thrust-to-weight ratio, specific fuel consumption, pressure loss and pattern factor.

“Address to Public Council”: The Additions to Marston’sThe Malcontent, the King’s Men’s Repertory, and Early Modern Theatrical Economics

“Address to Public Council”: The Additions to Marston’s<i>The Malcontent</i>, the King’s Men’s Repertory, and Early Modern Theatrical Economics

R. Judah he-Hasid or R. Moshe Zaltman: who proposed that Torah verses were written after the time of Moses?

In the year 1975, I.S. Lange published an anthology of commentaries entitled Commentaries on the Torah of R. Judah he-Hasid . In three of the commentaries included in this collection, the exegete suggests that the Torah includes post-Mosaic interpolations. In an additional passage, the commentator maintains that David removed psalms from the Torah and incorporated them into the book of Psalms. Some Orthodox rabbis found these comments to be heretical and rejected the possibility that they were written by R. Judah he-Hasid. In response to this criticism, Lange published a censored edition of the book, without the controversial sections. These four passages have been subject to considerable academic inquiry. On the basis of these commentaries, it is commonly maintained that R. Judah he-Hasid had idiosyncratic views about the composition of the Hebrew Bible. In this article, I argue that the controversial notes were not written by R. Judah he-Hasid but rather by his son, R. Moshe Zaltman.

Left ventricular assist devices: a kidney's perspective.

The left ventricular assist device (LVAD) has become an established treatment option for patients with refractory heart failure. Many of these patients experience chronic kidney disease (CKD) due to chronic cardiorenal syndrome type II, which is often alleviated quickly following LVAD implantation. Nevertheless, reversibility of CKD remains difficult to predict. Interestingly, initial recovery of GFR appears to be transient, being followed by gradual but significant late decline. Nevertheless, GFR often remains elevated compared to preimplant status. Larger GFR increases are followed by a proportionally larger late decline. Several explanations for this gradual decline in renal function after LVAD therapy have been proposed, yet a definitive answer remains elusive. Mortality predictors of LVAD implantation are the occurrence of either postimplantation acute kidney injury (AKI) or preimplant CKD. However, patient outcomes continue to improve as LVAD therapy becomes more widespread, and adverse events including AKI appear to decline. In light of a growing destination therapy population, it is important to understand the cumulative effects of long-term LVAD support on kidney function. Additional research and passage of time are required to further unravel the intricate relationships between the LVAD and the kidney.

Evolution of and evolutionary relationships between extant vaccinia virus strains.

Although vaccinia virus (VACV) was once used as a vaccine to eradicate smallpox on a worldwide scale, the biological origins of VACV are uncertain, as are the historical relationships between the different strains once used as smallpox vaccines. Here, we sequenced additional VACV strains that either represent relatively pristine examples of old vaccines (e.g., Dryvax, Lister, and Tashkent) or have been subjected to additional laboratory passage (e.g., IHD-W and WR). These genome sequences were compared with those previously reported for other VACVs as well as other orthopoxviruses. These extant VACVs do not always cluster in simple phylogenetic trees that are aligned with the known historical relationships between these strains. Rather, the pattern of deletions suggests that all existing strains likely come from a complex stock of viruses that has been passaged, distributed, and randomly sampled over time, thus obscuring simple historical or geographic links. We examined surviving nonclonal vaccine stocks, like Dryvax, which continue to harbor larger and now rare variants, including one that we have designated "clone DPP25." DPP25 encodes genes not found in most VACV strains, including an ankyrin-F-box protein, a homolog of the variola virus (Bangladesh) B18R gene which we show can be deleted without affecting virulence in mice. We propose a simple common mechanism by which recombination of a larger and hypothetical DPP25-like ancestral strain, combined with selection for retention of critically important genes near the terminal inverted repeat boundaries (vaccinia virus growth factor gene and an interferon alpha/beta receptor homolog), could produce all known VACV variants. Smallpox was eradicated by using a combination of intensive disease surveillance and vaccination using vaccinia virus (VACV). Interestingly, little is known about the historical relationships between different strains of VACV and how these viruses may have evolved from a common ancestral strain. To understand these relationships, additional strains were sequenced and compared to existing strains of VACV as well as other orthopoxviruses by using whole-genome sequence alignments. Extant strains of VACV did not always cluster in simple phylogenetic trees based on known historical relationships between these strains. Based on these findings, it is possible that all existing strains of VACV are derived from a single complex stock of viruses that has been passaged, distributed, and sampled over time.

High throughput in vitro micropropagation of sugarcane (Saccharum officinarum L.) from spindle leaf roll segments: Cost analysis for agri-business industry

High throughput micropropagation (HTM) protocol in sugarcane through direct shoot regeneration comprising five stages, was developed for agri-business industry. The distinction of the protocol lies in direct adventitious shoot regeneration without an intervening callus phase and liberty over the number of subculture passages leading to high rates of synchronous plant production. Stage 0 dealt with selection and maintenance of field-grown stock plants; Stage I marked the initiation of in vitro propagation on culture initiation medium using spindle leaf roll segments from apical tops; Stage II involved five passages (of 14 days each) on shoot multiplication medium followed by one additional passage (of 10 days) on shoot multiplication-cum-elongation medium. The shoot multiplication rates ranged from 4 to 25-fold in CoPb 91 and CoJ 83 cultivars, respectively; Stage III involved in vitro rooting on root induction medium and hardening for 14 days at each step; Stage IV comprised separation of clumps into single plantlets and transfer to soil. The complete plants were produced in 157 days with 97 percent survival rate. The fidelity of the protocol for agri-business industry was tested by producing approximately 0.1 million saleable HTM sugarcane plants in a small scale (150 m2) tissue culture unit. The recurring (energy, labour, media, culture container, transplanting material) and non-recurring (laboratory infrastructure, glasshouse, equipment) parameters accounted for 75.56 and 24.44 percent production costs, respectively. The low-cost options for media, culture containers and transplanting material were incorporated and cost per plant was calculated to be US$ 0.13. The HTM protocol was adopted by two sugar mills, and the HTM plants produced in our tissue culture unit were sold for seed multiplication to eight sugar mills and 430 farmers throughout North India.