Disposable Bags Research Articles

From leaf explants to hanging rooted plantlets in a mist reactor

A mist bioreactor using a disposable bag as a culture chamber was used to propagate single nodal explants of Artemisia annua into rooted plantlets that were ready for soil using a single batch (one-step) culture. To vertically scale plant growth inside the mist reactor, poly-l-lysine (PLL)-coated 70 µm nylon mesh and solid polypropylene sheeting were used for explant attachment. Both manually chopped and blender-chopped (blenderized) shoot tissues were attached to PLL-coated substrates. Compared to blenderized shoots, manually chopped tissues were larger with better attachment to PLL-coated substrates. Regardless of substrates or explant preparation method, 80–95 % of initially attached shoot tissues remained attached to PLL-coated surfaces after being misted with culture medium for 24 h. New shoot proliferation increased about tenfold as the size of blenderized shoot tissue increased. To reduce callusing during shoot proliferation and thus stimulate root initiation, original shooting medium was reduced to half strength of phytohormone and Murashige and Skoog salts. The duration of shoot proliferation was also reduced from 2 to 1 week. Original rooting medium was then further improved with NAA or IBA. After successive shooting, rooting and in vitro acclimatization, the nodal explants attached to PLL-coated hanging strips and developed into fully rooted plantlets in the mist reactor. Although most of the large rooted plantlets detached from the hanging strips by the time of harvest, they had fully functional stomata and were later successfully established in the soil, suggesting this “hanging garden” technology may prove useful for micropropagation.

Bans vs. Fees: Disposable Carryout Bag Policies and Bag Usage

AbstractUsing observational data on consumer carryout bag usage, we measure the effects of disposable bag policies on disposable and reusable bag demand. Our results show that plastic bag bans coupled with paper bag fees decrease total disposable bag demand but lead to significant increases in paper bag consumption. We compare our results to a study on bag fees and find that both policies lead to similar increases in reusable bag usage. However, the success of bans versus fees in discouraging disposable bag usage is contingent upon the types and prices of bags that stores choose to sell in lieu of disposable plastic.

BYOB: How Bringing Your Own Shopping Bags Leads to Treating Yourself and the Environment

As concerns about pollution and climate change become more mainstream, the belief that shopping with reusable grocery bags is an important environmental and socially conscious choice has gained prevalence. In parallel, firms have joined policy makers in using a variety of initiatives to reduce the use of disposable plastic bags. However, little is known about how these initiatives might alter other elements of consumers' in-store behavior. Using scanner panel data from a single California location of a major grocery chain, and controlling for consumer heterogeneity, the authors demonstrate that bringing one's own bags increases purchases of not only environmentally friendly organic foods but also indulgent foods. They use experimental methods to further explore the expression of these effects and to consider the effects of potential moderators, including competing goals and store policies. The findings have implications for decisions related to product pricing, placement and assortment, store layout, and the choice of strategies employed to increase the use of reusable bags.

A Feasibility on SRF and Mixing Ratio for Energy Efficiency with Bulky Waste and Disposable Plastic Bag

A Feasibility on SRF and Mixing Ratio for Energy Efficiency with Bulky Waste and Disposable Plastic Bag

Why Washington, DC, Is “One of the Best Places to Live in the World”: An Interview with Tommy Wells, Director of the District Department of the Environment

Tommy Wells is the director of the District Department of the Environment (DDOE). He first came to the District in 1983 after receiving his bachelor’s degree from the University of Alabama and a master’s degree in social work from the University of Minnesota. Wells focused on child welfare issues for over two decades, first at the city’s child protective services agency and then as director of the Consortium for Child Welfare, a coalition of nonprofit organizations advocating for children, youth, and families in the District. During that time he attended law school at night, receiving his degree from the Catholic University of America in 1991.Wells first held elected office in 1995, as a member of the Advisory Neighborhood Commission in Ward 6. Following a stint on the DC State Board of Education, he ran for city council, where he represented Ward 6 for eight years. During his time on the council, Wells sat on the committees responsible for legislation affecting the environment, health, human services, and transportation. In 2014, he ran in the Democratic primary for mayor, finishing third to current mayor Muriel Bowser and then-mayor Vincent Gray.On March 18, Wells spoke with Christine Mellen of Policy Perspectives at his office. Their conversation touched on topics such as the District’s disposable bag fee, energy and the environment, and the District’s streetcar system.

Minipool caprylic acid fractionation of plasma using disposable equipment: a practical method to enhance immunoglobulin supply in developing countries.

BackgroundImmunoglobulin G (IgG) is an essential plasma-derived medicine that is lacking in developing countries. IgG shortages leave immunodeficient patients without treatment, exposing them to devastating recurrent infections from local pathogens. A simple and practical method for producing IgG from normal or convalescent plasma collected in developing countries is needed to provide better, faster access to IgG for patients in need.Methodology/Principal FindingsIgG was purified from 10 consecutive minipools of 20 plasma donations collected in Egypt using single-use equipment. Plasma donations in their collection bags were subjected to 5%-pH5.5 caprylic acid treatment for 90 min at 31°C, and centrifuged to remove the precipitate. Supernatants were pooled, then dialyzed and concentrated using a commercial disposable hemodialyzer. The final preparation was filtered online by gravity, aseptically dispensed into storage transfusion bags, and frozen at <-20°C. The resulting preparation had a mean protein content of 60.5 g/L, 90.2% immunoglobulins, including 83.2% IgG, 12.4% IgA, and 4.4% IgM, and residual albumin. There was fourfold to sixfold enrichment of anti-hepatitis B and anti-rubella antibodies. Analyses of aggregates (<3%), prekallicrein (5-7 IU/mL), plasmin (26.3 mU/mL), thrombin (2.5 mU/mL), thrombin-like activity (0.011 U/g), thrombin generation capacity (< 223 nM), and Factor XI (<0.01 U/mL) activity, Factor XI/XIa antigen (2.4 ng/g) endotoxin (<0.5 EU/mL), and general safety test in rats showed the in vitro safety profile. Viral validation revealed >5 logs reduction of HIV, BVDV, and PRV infectivity in less than 15 min of caprylic acid treatment.Conclusions/Significance90% pure, virally-inactivated immunoglobulins can be prepared from plasma minipools using simple disposable equipment and bag systems. This easy-to-implement process could be used to produce immunoglobulins from local plasma in developing countries to treat immunodeficient patients. It is also relevant for preparing hyperimmune IgG from convalescent plasma during infectious outbreaks such as the current Ebola virus episode.

MICROBIOLOGY OF RAW MATERIALS USED FOR CONFECTIONARY PRODUCTION

The aim of our study was to evaluate the microbiological quality of raw materials used for preparation of confectionery products. For microbiological evaluation total count of bacteria, mesophilic aerobic bacteria, coliform bacteria, yeast and microscopic filamentous fungi in samples of raw materials used in the manufacture and creams of confectionery products were detected. In addition to these groups of microorganisms the presence of pathogenic microorganisms Salmonella spp. and Staphylococcus aureus in creams was monitored. Products are assessed according to the limit values of the number of microorganisms defined in the Codex Alimentary of the Slovak Republic. For microbiological analysis of confectionery products, sampling of components of confectionary products and cream was carried out according to current health regulations and altogether 65 samples of components and creams were collected: 10 samples of raw materials sugar, 10 samples of flour, 10 samples of butter and 10 samples of eggs, 5 samples of butter yolk from cream-filled disposable bag without rum addition, 5 samples of butter yolk from cream-filled disposable bag with rum addition, 5 samples of cream-filled multiple use paid bag, 5 samples of cream-filled newly purchased paid bag, 5 samples of Venček corpus and 5 samples of the French cubes corpus. From raw material the highest TBC (2.65log CFU) was in flour, but the lowest in sugar (1.35 log CFU), the highest years counts was found on flour (2.42), but lowest in butter (1.18), while wasn’t in egg. In samples of creams and corpus were increased occurrence of yeast, coliform bacteria. Salmonella spp. and Staphylococcus aureus weren’t isolated from any tested sample.

Energy Efficiency, Sustainable Development and Natural Resources Conservation: Multicriteria Life-cycle Analysis of Daily-Used Materials with a Long-term Vision

This paper explores the role of energy use in sustainable development and the potential sources to increase energy efficiency during the whole life - cycle of any material and its production process . It is the first paper of a research project exploring a decision making model for a multidisciplinary problem in nature. It deals with Multicriteria decisio n making for plastic materials used in a day to day basis. We analyze plastic materials used to manufacture disposable polyethylene bags among other materials that can be used for their substitution. We are also interested in plastic (polyethylene Terephta late or PET) bottles and its possible substitutes. Sustainability considers the concept of exergy loss, Green House Gases emissions, real energy

Comparison of the disposable bag replacement time of biliary T-tube drainage

Objective To compare the replacement time of normal and anti-reflux disposable drainage bag for the risk of infection in patients with biliary T-tube drainage, and to explore the most suitable frequency which patients can get the most benefits.Methods Totals of 144 patients with routine T-tube drainage and the negative culture results of biliary bile were randomly divided into three groups by random number table method. Group A and group B used ordinary disposable drainage bag, while group A changed it every day and group B changed it once a week.Group C used the disposable anti-reflux drainage bag and replaced it once a week.The extraction of bile from drainage bag was used to do the bacteriological monitoring and the results were compared among the three groups.Results There was no significant difference in the bacterial culture among the three groups at the 5th day and 10th day after the drainage （χ2 =0.51, 1.64, respectively;P 〉0.05）.The replacement frequency of the three groups were statistically significant （F=1 379.17,P〈0.01）.The cost of changing drainage bag in three groups had statistically significant difference （ F =229.19, P 〈0.01 ）. Conclusions Patients with biliary T-tube, who use the ordinary disposable bags and change it weekly have similar infection rate as the anti-reflux drainage bag, what’ s more, it can reduce the costs of patients and the workload of nurses. Key words: Biliary track; T-tube drainage; Disposable drainage bag; Replacement time

Scaled-up manufacturing of recombinant antibodies produced by plant cells in a 200-L orbitally-shaken disposable bioreactor.

Tobacco BY-2 cells have emerged as a promising platform for the manufacture of biopharmaceutical proteins, offering efficient protein secretion, favourable growth characteristics and cultivation in containment under a controlled environment. The cultivation of BY-2 cells in disposable bioreactors is a useful alternative to conventional stainless steel stirred-tank reactors, and orbitally-shaken bioreactors could provide further advantages such as simple bag geometry, scalability and predictable process settings. We carried out a scale-up study, using a 200-L orbitally-shaken bioreactor holding disposable bags, and BY-2 cells producing the human monoclonal antibody M12. We found that cell growth and recombinant protein accumulation were comparable to standard shake flask cultivation, despite a 200-fold difference in cultivation volume. Final cell fresh weights of 300-387 g/L and M12 yields of ∼20 mg/L were achieved with both cultivation methods. Furthermore, we established an efficient downstream process for the recovery of M12 from the culture broth. The viscous spent medium prevented clarification using filtration devices, but we used expanded bed adsorption (EBA) chromatography with SP Sepharose as an alternative for the efficient capture of the M12 antibody. EBA was introduced as an initial purification step prior to protein A affinity chromatography, resulting in an overall M12 recovery of 75-85% and a purity of >95%. Our results demonstrate the suitability of orbitally-shaken bioreactors for the scaled-up cultivation of plant cell suspension cultures and provide a strategy for the efficient purification of antibodies from the BY-2 culture medium.

Isolation and characterisation of fungal communities associated with degradation and growth on the surface of poly(lactic) acid (PLA) in soil and compost

Polylactic acid (PLA) is a compostable biopolymer that is currently used for short shelf-life product containers and waste disposal bags. At temperatures at or above its glass transition temperature (55°–62 °C), PLA in the presence of water undergoes chemical hydrolysis, progressively reducing polymer molecular weight and ultimately releasing lactic acid. The role of microorganisms in this process has largely focused on bacteria, with few studies investigating potential role of fungi. In this study, PLA (96% l-isomer) coupons were buried in soil or compost and incubated at 25 °C or 50 °C for up to 8 weeks. Physical disintegration of the coupons was seen only when incubated at 50 °C in both soil and compost. The principle fungal species identified from PLA coupons at 25 °C included Mortierella sp., Doratomyces microsporus, Fusarium solani, Fennellomyces sp., Aspergillus fumigatus, Verticillium sp., Lecanicillium saksenae, Cladosporium sp. and Trichoderma sp. Only Thermomyces lanuginosus and A. fumigatus were isolated from PLA at 50 °C and both were shown to cause significant PLA biodegradation as determined by tensile strength loss compared to abiotic controls. Terminal restriction fragment length polymorphism (TRFLP) demonstrated strong selection of specific fungal species from soil and compost communities on PLA the surface and a greater species diversity than could be detected by conventional culturing techniques.

Soil sampling and analytical strategies for mapping fallout in nuclear emergencies based on the Fukushima Dai-ichi Nuclear Power Plant accident

The Fukushima Dai-ichi Nuclear Power Plant (FDNPP) accident resulted in extensive radioactive contamination of the environment via deposited radionuclides such as radiocesium and 131I. Evaluating the extent and level of environmental contamination is critical to protecting citizens in affected areas and to planning decontamination efforts. However, a standardized soil sampling protocol is needed in such emergencies to facilitate the collection of large, tractable samples for measuring gamma-emitting radionuclides. In this study, we developed an emergency soil sampling protocol based on preliminary sampling from the FDNPP accident-affected area. We also present the results of a preliminary experiment aimed to evaluate the influence of various procedures (e.g., mixing, number of samples) on measured radioactivity. Results show that sample mixing strongly affects measured radioactivity in soil samples. Furthermore, for homogenization, shaking the plastic sample container at least 150 times or disaggregating soil by hand-rolling in a disposable plastic bag is required. Finally, we determined that five soil samples within a 3 m × 3-m area are the minimum number required for reducing measurement uncertainty in the emergency soil sampling protocol proposed here.

Providing anti-retroviral therapy in the context of self-perceived stigma: a mixed methods study from Tanzania.

Adherence to anti-retroviral treatment (ART) has been a significant step towards improving quality of life among people living with HIV. However, stigma has been described to influence adherence to ART. A cross-sectional mixed methods study was conducted to explore factors related to stigma and perceived influence of stigma on adherence to treatment amongst ART-prescribed patients and health care providers, respectively in Tanzania. Stigma was assessed through interviewer administered survey among 295 patients. The results from patients showed that 279/295 (95%) were satisfied with the services provided at the Care and Treatment Centres (CTCs). The set up of CTCs 107/295 (36%), and queuing at the CTCs 88/295 (30%) were associated with stigma (P < 0.001). The perceived influence of stigma on adherence to ART was assessed using focus group discussions (FGDs) of 33 health care providers (HCPs). Through FGDs, HCPs perceived the set up of CTCs as friendly yet violated confidentiality. The HCPs reported that ART-prescribed patients hide identifiable cards to avoid being recognised by other people. Some patients were reported to rush to avoid familiar faces, and due to the rush they picked wrong medicines. Also some patients were reported to throw away manufacturers' box with dosage instructions written on the box, resulting in use of doses contrary to the prescriptions. We conclude that despite the fact that most patients were satisfied with the services provided at the CTCs, it is important that HCPs provide dosage instructions on another piece of paper or use disposable bags. A common dispensing window for all patients regardless of the diagnosis may be useful to minimize stigma. Also HCPs may introduce appointment system to avoid long queue at the CTCs.

Correlation between the replacing time of plasma drainage bag and the bacterial growth rate

Objective To explore the correlation between the replacing time of plasma drainage bag and the bacterial growth rate among orthopaedic surgery patients , and to control or reduce the risk of catheter-related infection.Methods The convenient sampling method was used to select 100 patients with postoperative serum drainage tube and disposable drainage bag , from September 2012 to August 2013.Patients were randomly divided into the control group （group A, 33 cases） and the observation group （group B, 32 cases; group C, 35 cases）.The amount of drainage were recorded every 24 hours of the three groups of patients , and then the drainage bags were emptied .Group A replaced the drainage bag regularly every day; Group B replaced the drainage bag every 48 hours;Group C did not replace the drainage bag .The outside mouth of the drainage tube and the drainage liquid were taken to bacterial culture . The positive rate of bacterial culture , patients’postoperative body temperature and the rate of incision infection were compared within groups .Results Twenty-four hours after operation, the results of bacterial culture among three groups were negative .However, after 48 hours, six patients （18.1%） in group A, two patients （6.4%） in group B showed positive results of the bacterial culture .None of the patients in group C had the positive results .The comparison among three groups was statistically significant （χ2 =10.32,P〈0.05）.After 72 hours, eight patients （24.2%） in group A, two patients （6.4%） in group B showed positive results of the bacterial culture .None of the patients in group C had the positive results.The comparison among three groups was statistically significant （χ2 =11.82,P〈0.05）. The incidence of incision infection of group A , group B and group C were 15.1%（5/33）, 6.3%（2/32） and＆amp;nbsp;0.0%（0/35）, respectively.There was a significant difference among three groups （χ2 =11.18,P〈0.05）.The body temperature of the three groups showed no significant difference （F=7.53,P〉0.05）.Conclusions With the increase of drainage bag replacement frequency , the positive rate of bacterial culture , as well as the drainage bag and drainage tube mouth bacteria culture positive rate increase .Therefore, for orthopaedic patients , the plasma drainage tube and disposable drainage bag should be emptied every 24 hours after the measurement of drainage liquid, but do not have to be replaced every 48 hours or every 24 hours. Key words: Infection; Postoperative care; Replacing time; Plasma drainage bag; Drainage tube; Bacterial growth rate

SACOLAS RETORNÁVEIS: UMA ALTERNATIVA PARA REDUÇÃO DO IMPACTO DE LARGA ESCALA CAUSADO PELA ELIMINAÇÃO IRREGULAR DE SACOLAS DESCARTÁVEIS

This study aimed to determine the receptivity and acceptance of customers in a supermarket chain in relation to the adoption of reusable bags as a strategy for environmental marketing. Therefore, we conducted a case study by implementing a structured questionnaire. The questionnaire was administered to a non-probability sample of 150 customers of a supermarket chain, located in the town of Santa Maria, RS. The results were positive regarding customer acceptance of replacement of disposable bags for reusable bags, however, despite the acceptance is high, adherence is still low, since most customers forget to take reusable bags when they go shopping. This result demonstrates the need to increase investments in environmental education, to raise awareness about the need and importance of protecting and preserving the environment.

Improved production of recombinant human Fas ligand extracellular domain in Pichia pastoris: yield enhancement using disposable culture-bag and its application to site-specific chemical modifications.

BackgroundA useful heterologous production system is required to obtain sufficient amounts of recombinant therapeutic proteins, which are often necessary for chemical characterization and engineering studies on the development of molecules with improved properties. Human Fas ligand extracellular domain (hFasLECD) is an agonistic death ligand protein that has potential applications for medical purposes. Site-specific chemical modifications can provide a powerful means for the development of engineered proteins with beneficial functions. This study aimed to enhance the yield of hFasLECD using a Pichia pastoris secretory expression system suitable for efficient production on a small laboratory scale, and further to provide procedures for its site-specific chemical modification without impairing the biological functions based on the developed production system.ResultsA convenient cultivation system using a disposable plastic bag provided a three-fold increase in purification yield of tag-free hFasLECD as compared with the conventional system using a baffled glass flask. The system was further applied to the production of a mutant, which contains an additional reactive cysteine residue in the N-terminal tag-sequence region. Site-specific conjugations and cross-linking without impairing biological functions were achieved by reaction of the mutant hFasLECD with single maleimide group containing compounds and a linear polyethylene glycol derivative containing two maleimide groups at either end, respectively. All purified tag-free and chemically modified hFasLECDs showed an evident receptor binding activity in co-immunoprecipitation experiments mediated by wild-type and N-glycosylation site deficient mutant human Fas receptor extracellular domain derivatives. An N-Ethylmaleimide conjugated hFasLECD derivative demonstrated a significant cytotoxic activity against human HT-29 colorectal cancer cells.ConclusionsA new, efficient cultivation system for enhanced secretory production of hFasLECD using P. pastoris and an effective strategy for site-specific chemical modifications of hFasLECD were devised. The results obtained constitute the basis for biomedical applications including developments of novel therapeutic proteins and diagnostic tools targeted to related diseases and their biomarkers.

A cytotoxic leachable compound from single‐use bioprocess equipment that causes poor cell growth performance

A current trend in the production of biopharmaceuticals is the replacement of fixed stainless steel fluid-handling units with disposable plastic bags. Such single-use systems (SUS) offer numerous advantages, but also introduce a new set of materials into the production process and consequently expose biomanufacturers to a new set of risks related to those materials, not to mention reliance on an entirely new supply chain. In the course of developing and conducting a cell-growth-based test for suitability of disposable plastic components destined for use in cell culture operations, we discovered that the cytotoxic compound bis(2,4-di-tert-butylphenyl)phosphate (bDtBPP) leaches out of certain bags and into cell culture media in concentrations that are deleterious to cell growth. Specifically, media held in certain bags for several days at 37°C was found to contain bDtBPP, and use of those held-media samples in cell growth experiments provides data that overlap neatly with cell growth experiments using media spiked directly with bDtBPP, proving that bDtBPP leaching is responsible for the reduced growth attributable to those SUS bags. Overall, this issue represents a risk to the production of biopharmaceuticals in SUS, a risk that must be managed by diligent collaboration among companies along the entire supply chain for SUS components.

Trocar Size Selection for Trans-Trocar Appendix Removal in Laparoscopic Appendectomy

Purpose: Many disposable laparoscopic instruments are used in laparoscopic surgery. There are several disadvantages in use of disposable laparoscopic instruments, including cost ineffectiveness, environmental contamination, and resource recycling. In addition, a disposable specimen bag has been used in laparoscopic appendectomy. Use of the trans-trocar appendix removal technique provides several advantages compared to use of a specimen bag. Therefore, the aim of this study is to analyze the adequate trocar size for trans-trocar appendix removal. Methods: A total of 62 patients undergoing appendectomy between June 2012 and September 2012 were identified. After appendectomy, we performed a trans-trocar test using removed appendix specimen and trocars. Three different types of trocars were used for the trans-trocar test, 5 mm, 12 mm, and 11 mm Xcel ® (Ethicon). We analyzed the success rate. Results: Significant relationships were observed between maximal specimen diameter and body mass index. When BMI was below 20, the success rate of the 11 mm trocar was 86% in the trans-trocar appendix removal test. W hen BMI was between 20 and 25, the success rate of the 12 mm trocar was 71%, but that of 11 mm was 57%. When BMI was above 25, the success rate of the 15 mm trocar was 62%. Conclusion: Although this study had many limitations, a large-sized trocar was needed for trans-trocar appendix removal in more body mass index. When BMI is below 20, an 11 mm trocar is recommended in trans-trocar appendix removal. When BMI is between 20 and 25, a 12 mm trocar is. When BMI is above 25, a 15 mm trocar and a disposable specimen bag are recommended. Further continuous study will be needed for analysis of clinical outcome.

Strategy for selecting disposable bags for cell culture media applications based on a root‐cause investigation

The use of disposable bags for cell culture media storage has grown significantly in the past decade. Some of the key advantages of using disposable bags relative to non-disposable containers include increased product throughput, decreased cleaning validation costs, reduced risk of cross contamination and lower facility costs. As the scope of use of disposable bags for cell culture applications increases, problematic bags and scenarios should be identified and addressed to continue improving disposables technologies and meet the biotech industry's needs. In this article, we examine a cell culture application wherein media stored in disposable bags is warmed at 37°C before use for cell culture operations. A problematic bag film was identified through a prospective and retrospective cell culture investigation. The investigation provided information on the scope and variation of the issue with respect to different Chinese hamster ovary (CHO) cell lines, cell culture media, and application-specific parameters. It also led to the development of application-specific test methods and enabled a strategy for disposable bag film testing. The strategy was implemented for qualifying an alternative bag film for use in our processes. In this test strategy, multiple lots of 13 bag film types, encompassing eight vendors were evaluated using a three round, cell culture-based test strategy. The test strategy resulted in the determination of four viable bag film options based on the technical data. The results of this evaluation were used to conclude that a volatile or air-quenched compound, likely generated by gamma irradiation of the problematic bag film, negatively impacted cell culture performance.

From cells to embryos to rooted plantlets in a mist bioreactor

A mist bioreactor using a disposable bag as culture chamber was used to propagate carrot embryogenic cells into rooted plantlets. The best operating configuration was akin to a vertical hanging garden using 50–90 μm nylon mesh for explant attachment. Cells spray inoculated into the reactor were 51.2 % viable. Misting cycle and aeration conditions were studied and showed that under the same hourly volumetric nutrient feed and 0 VVM, embryo development in the reactor was best using a 0.3 min on/2.7 min off misting cycle, yielding about 23 % post heart stage embryos. Compared to 0 VVM, 3 % CO2 enrichment improved embryo development in reactor culture. Spray inoculated cells also attached to several vertically hung poly-l-lysine coated strips and then developed in situ into embryos. Cell attachment was significantly improved when they were suspended in salt-free sucrose solution during spray inoculation. Almost 90 % of the originally attached cells remained on the nylon mesh 24 h later after spraying with B5 medium in the mist reactor. Strip grown embryos had the same post heart stage ratio but shorter overall length compared to those developed on a horizontal platform. Young plantlets developed uniformly up and down the hanging strips and did not detach after 3 weeks of culture suggesting this technology may prove useful for improving micropropagation.