Initial Cell Count Research Articles

Influence of Lens Status on Outcomes of Descemet Membrane Endothelial Keratoplasty.

To evaluate whether lens status influences clinical outcomes of Descemet membrane endothelial keratoplasty (DMEK) in patients who underwent DMEK in phakia, pseudophakia, or combined with cataract surgery (triple). A retrospective review of 139 eyes (107 patients) with symptomatic Fuchs' dystrophy or secondary corneal edema was performed. Patients were divided into 3 groups: 1) DMEK + cataract surgery, 2) DMEK in pseudophakia, and 3) DMEK in phakia. Of 139 DMEK surgeries, 61 were DMEK triple (43.9%), 55 pseudophakic DMEK (39.6%), and 23 phakic DMEK (16.5%) procedures. Rebubble occurred in 31 of 134 eyes (23.1%), of which 17 were in eyes after DMEK triple (28.3%), 10 after pseudophakic DMEK (19.6%), and 4 after phakic DMEK (17.4%). There was no statistically significant difference in rebubble rates depending on lens status (P = 0.548). There was no statistically significant difference in best corrected visual acuity (BCVA) between eyes that underwent rebubble versus not (P = 0.854). BCVA in patients post-DMEK was 20/25 or better in 81% of cases, and mean BCVA was 20/23. Donor age (P = 0.889), graft cell count (P = 0.525), and graft preparation technique (P = 0.769) were not predictive of rebubbling. Our study demonstrates no difference in DMEK outcome with regard to rebubble rate whether the procedure is performed in combination with cataract surgery, in pseudophakia, or in phakia. Furthermore, donor age, initial graft cell count, and graft preparation technique did not impact rebubble rate. BCVA was also not affected by the need for rebubbling.

UV-C treatment of grape must: Microbial inactivation, toxicological considerations and influence on chemical and sensory properties of white wine

UV-C was investigated to inactivate microorganisms in grape must. The aim was to evaluate the effect of UV-C on wine-related yeasts under winery conditions, to investigate the toxicological potential and to analyze the effects on chemical and sensory wine properties. Metschnikowia pulcherrima showed the highest UV-C tolerance, Saccharomyces cerevisiae the lowest. With an initial cell count of 106 cfu/mL, M. pulcherrima required a UV-C dose of >1.2 kJ/L, S. cerevisiae <0.8 kJ/L for inactivation. Inactivation efficacy decreased with higher must turbidity (26.7–144.5 NTU), optical density and degree of microbial load (104–108 cfu/mL) suggesting a shadowing effect of individual microbes. UV-C treatment did not impact mutagenicity as tested by the Ames test. A decrease in dissolved oxygen and caftaric acid content indicated oxidative reactions. UV-C induced color bleaching (ΔE: 0–13) was also observed indicating a degradation of colored pigments. GC × GC–MS analysis revealed a decrease of β-damascenone and linalool content in wines made from UV-C treated must. Descriptive analysis by a trained sensory panel showed that off-flavors did not occur at doses that were relevant for microbial inactivation (between 1.0 and 3.0 kJ/L). UV-C treatment is a non-thermal processing technique for the preservation of food. In this study, the applicability of UV-C treatment for the inactivation of spoilage yeast in grape must was demonstrated in pilot scale. The treatment provided sufficient microbial inactivation without having toxicological impact as examined by the Ames test and without causing off-flavor formation in the wines using relevant doses.

Efficacy of Antimicrobial-Impregnated External Ventricular Drain Catheters: A Prospective, Randomized, Controlled Trial

AbstractBackground: Insertion of External Ventricular Drain (EVD) is associated with considerable risk of Cerebrospinal Fluid (CSF) infection which is a life-threatening complication. Infection occurs when microorganisms colonize along the catheter surface.Aim of Study: The aim of this study is to compare the safety & efficacy of EVD catheters impregnated with Clin-damycin & rifampin with those of standard control catheters for prevention of the catheter-related infections.Subjects and Methods: The prospective study was con-ducted on forty patients between April 2013 – November 2014. All patients were randomly assigned to undergo place-ment of an EVD with a catheter impregnated with Clindamycin & rifampin or a standard non-impregnated catheter (control group). Each group included twenty patients. Standard silicone catheter was the EVD used in both cohorts. Patients diagnosed with a known or suspected CSF infection were excluded from this study. Initial cell counts and cultures were obtained from the CSF, repeated at intervals (72 hours) and at time of Catheter removal. CSF infection rate and its impacton the clinical outcome were evaluated in both groups.Results: Among this series of forty patients with EVDs, three had an EVD-associated CSF infection in the control group giving an infection rate within this group of 15% per patient. Only one case in the Antibiotics-impregnated Catheter (AIC) group had CSF infection. Positive CSF cultures were obtained on day 10, 12 & 14 respectively. Organism isolated from CSF cultures was mainly gram-negative infection. AIC group showed a significant low infection rate compared to the uncoated one (p<0.001).Conclusion: The Catheters impregnated with the antibiotic combination of Clindamycin and rifampin appears to be safe and effective in reducing the risk of infectious complications associated with the placement of an EVD.

Efficacy of Decitabine Combined with Pre-Excitation Chemotherapy in the Treatment of Middle-Aged and Elderly MDS Transformed Acute Myeloid Leukemia

To investigate the efficacy of domestic decitabine (D) combined with pre-excitation chemotherapy consisted of Ara-c, THP and G-CSF(CTG) in treatment of middle-aged and elderly patients with MDS-transformed AML and prognosis-related factors. Seventy-six patients with MDS-transformed AML treated in our hospital from June 2013 to June 2015 were selected according to treatment regimens, 76 patients were divided into 2 groups: CTG group(36 cases) and D+CTG group(40 cases). The patients in CTG group received treatment with Ara-C, THP and G-CSF; the patients received the treatment with decitabine plus CTG. The patients in 2 groups all received 4 course treatment, then received maintaining treatment. The therapeutic efficacy and incidence of adverse reactions in 2 group were compared, at the same time, the risk factors affecting the prognos of patients treated with D+CTG were analyzed. There were no siginificant differences in age, sex, initial blood cell count, bone marrow blast ratio, disease types, chromosome karyotypes and FLT3-ITD gene mutation between 2 groups. The efficacy analysis showed that the efficacy of D+CTG was superior to CTG, ORR in D+CTG group was significantly higher than that in CTG group (72、52 vs 50%) (P<0.05), moreover, no significant differences in bone marrow inhibition digree infeetion, gastroinfestinal response and liver damage were found between 2 groups (P>0.05). The follow-np for 2 years showed that the median survival time in D+CTG group was significantly longer than that in CTG group (19.9 vs 11.0 months) (P<0.05). The multivariate analysis showed that the 1 course efficacy (RR=3.926, P=0.015) and FLT3-ITD gene mutation (RR=4.347, P=0.004) were independent risk factors affecting the efficacy of D+CTG treatment. The short-and long-term efficacy of domestic decitasine combined with preexcitation chenotherapy in treatment of middec-aged and eldery patients with MDS transformed AML is superior to single pre-excitation chenothrapy, moreover the incidence of adverse reactions did not increase. The 1 course efficacy and FLT-3 ITD gene mutation are the independent risk factors affecting the prognosis of patients. .

Extended live-tracking and quantitative characterization of wound healing and cell migration with SiR-Hoechst

Cell migration is essential to many life processes, including immune response, tissue repair, and cancer progression. A reliable quantitative characterization of the cell migration can therefore aid in the high throughput screening of drug efficacy in wound healing and cancer treatments. In this work, we report what we believe is the first use of SiR-Hoechst for extended live tracking and automated analysis of cell migration and wound healing. We showed through rigorous statistical comparisons that this far-red label does not affect migratory behavior. We observed excellent automated tracking of random cell migration, in which the motility parameters (speed, displacement, path length, directionality ratio, persistence time, and direction autocorrelation) obtained closely match those obtained from manual tracking. We also present an analysis framework to characterize the healing of a scratch wound from the perspective of single cells. The use of SiR-Hoechst is advantageous for the crowded environments in wound healing assays because as long as cell nuclei do not overlap, continuous tracking can be maintained even if there is cell-cell contact. In this paper, we report wound recovery based on the number of cells migrating into the wound over time, normalized by the initial cell count prior to the infliction of the wound. This normalized cell count approach is impervious to operator bias during the arbitration of wound edges and is also robust against variability that arises due to differences in the cell density of different samples. Additional wound healing characteristics were also defined based on the evolution of cell speed and directionality during healing. Not unexpected, the wound healing cells exhibited much higher tendency to maintain the same migratory direction in comparison to the randomly migrating cells. The use of SiR-Hoechst thus greatly simplified the automation of single cell and whole population analysis with high spatial and temporal resolution over extended periods of time.

Survival Outcome of Alternative Medicine Treatment for Newly Diagnosed Acute Leukemia in Children

Background: There is scarce information on the efficacy of alternative medicine (AM) alone as a treatment for newly diagnosed acute leukemia in children. We aimed to compare overall survival (OS) between children with newly diagnosed acute leukemia who received AM alone as the first-line treatment and those treated with conventional chemotherapy (CCT). Methods: Two-to-one nearest-neighbor propensity score-matching using sex, initial white blood cell count, phenotype of leukemia, and period of diagnosis was performed on 184 patients who received CCT and 92 who received AM alone after being diagnosed with leukemia. A multivariable Cox proportional-hazards regression model was then applied to assess the effect of treatment on OS after adjusting for potential confounders. Hazard ratios (HR) and 95% confidence intervals (CI) are provided. Results: After adjusting for initial white cell count and subtype of leukemia, children treated with AM alone had worse OS (HR 5.14, 95% CI 3.75–7.04) than those given CCT. The 5-year OS rate for newly diagnosed acute leukemia treated with AM medicine alone was 0%. Conclusion: AM without CCT is associated with poorer survival when compared with CCT.

Antibiotic loaded calcium sulfate bead and pulse lavage eradicates biofilms on metal implant materials in vitro.

Pulse lavage (PL) debridement and antibiotic loaded calcium sulfate beads (CS-B) are both used for the treatment of biofilm related periprosthetic joint infection (PJI). However, the efficacy of these alone and in combination for eradicating biofilm from orthopaedic metal implant surfaces is unclear. The purpose of the study was to understand the efficacy of PL and antibiotic loaded CS-B in eradicating bacterial biofilms on 316L stainless steel (SS) alone and in combination in vitro. Biofilms of bioluminescent strains of Pseudomonas aeruginosa Xen41 and a USA300 MRSA Staphylococcus aureus SAP231 were grown on SS coupons for 3 days. The coupons were either, (i) debrided for 3 s with PL, (ii) exposed to tobramycin (TOB) and vancomycin (VAN) loaded CS-B for 24 h, or (iii) exposed to both. An untreated biofilm served as a control. The amount of biofilm was measured by bioluminescence, viable plate count and confocal microscopy using live/dead staining. PL alone reduced the CFU count of both strains of biofilms by approximately 2 orders of magnitude, from an initial cell count on metal surface of approximately 109 CFU/cm2 . The antibiotic loaded CS-B caused an approximate six log reduction and the combination completely eradicated viable biofilm bacteria. Bioluminescence and confocal imaging corroborated the CFU data. While PL and antibiotic loaded CS-B both significantly reduced biofilm, the combination of two was more effective than alone in removing biofilms from SS implant surfaces. © 2018 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:2349-2354, 2018.

High-Throughput Quantification of Bacterial-Cell Interactions Using Virtual Colony Counts

The quantification of bacteria in cell culture infection models is of paramount importance for the characterization of host-pathogen interactions and pathogenicity factors involved. The standard to enumerate bacteria in these assays is plating of a dilution series on solid agar and counting of the resulting colony forming units (CFU). In contrast, the virtual colony count (VCC) method is a high-throughput compatible alternative with minimized manual input. Based on the recording of quantitative growth kinetics, VCC relates the time to reach a given absorbance threshold to the initial cell count using a series of calibration curves. Here, we adapted the VCC method using the model organism Salmonella enterica sv. Typhimurium (S. Typhimurium) in combination with established cell culture-based infection models. For HeLa infections, a direct side-by-side comparison showed a good correlation of VCC with CFU counting after plating. For MDCK cells and RAW macrophages we found that VCC reproduced the expected phenotypes of different S. Typhimurium mutants. Furthermore, we demonstrated the use of VCC to test the inhibition of Salmonella invasion by the probiotic E. coli strain Nissle 1917. Taken together, VCC provides a flexible, label-free, automation-compatible methodology to quantify bacteria in in vitro infection assays.

Case 2: A Patient with Trisomy 21, Transient Abnormal Myelopoiesis, and Pneumoperitoneum

A 34-week-gestation male infant is born appropriate for gestational age to a 41-year-old gravida 3, para 3 woman. The patient’s mother had received limited prenatal care starting at 15 weeks, and is found to have a positive result on maternal serum α-fetoprotein screening test and an amniocentesis that confirms trisomy 21 (47,XY+21 karyotype) with no evidence of translocation or mosaicism. The infant is born at an outside hospital and has physical features characteristic of trisomy 21. He is transferred to our facility for respiratory distress and multiple episodes of apnea and bradycardia on the day of birth. Sepsis is ruled out on admission. His initial complete blood cell count is significant for a white blood cell count of 15,300/μL (15.3×109/L) with 25% blasts. He is also noted to have an enlarged liver that is 1 cm below the costal margin. Peripheral blood smear and flow cytometry findings are consistent with a diagnosis of transient abnormal myelopoiesis (TAM). Pediatric hematology is consulted and recommends complete blood cell counts which are obtained regularly until day 22 after birth, when the TAM spontaneously resolves. The infant is able to receive full enteral feedings until he develops abdominal distention and irritability on day 12 after birth. An abdominal radiograph is notable for pneumoperitoneum concerning for an acute bowel perforation without signs of …

Leaks in fixed-ring banded sleeve gastrectomies: a management approach

BackgroundThe use of a Fobi ring to prevent pouch dilation is sometimes used in Roux-en-Y gastric bypass (RYGB). Recently, it has been extrapolated to laparoscopic sleeve gastrectomy (LSG) procedures by placing a fixed-ring band a few centimeters below the gastroesophageal junction (GEJ). ObjectivesWhat is the consequence if a patient develops a leak? SettingTertiary metropolitan referral center, Australia. MethodsOver 18 months, all patients with either a conventional LSG or a fixed-ring banded sleeve gastrectomy (BLSG) who presented with a proven leak complication were included. The management approaches along with the surgical, endoscopic, and percutaneous procedures used were examined. ResultsSix patients had a BLSG leak and 6 had a LSG leak. All patients had leak resolution. There was no significant difference in body mass index (BMI), time to leak, initial white cell count (WCC), and C-reactive protein (CRP) levels between the 2 groups. LSG patients required a median of 2 endoscopic procedures (range: 1–3). Stents were deployed in 3 patients. All BLSG patients required a single surgical intervention with laparoscopic washout, drainage, removal of band±feeding jejunostomy. One stent was deployed in 1 BLSG patient. BLSG leak resolution was found at 34±12 days versus 85±12 days in the LSG group (P< .05). ConclusionThe BLSG is a new modification of the sleeve gastrectomy procedure. This study presents a management strategy for leak resolution employed in BLSG patients. The presence of a foreign body as a persistent nidus of infection mandates band removal.

호박고구마 효소 분해물의 분무건조 분말 제조 및 물리화학적 품질특성

This study was conducted prepare spray-dried powder using pumpkin sweet potato hydrolysates and examine the physicochemical properties of the powder. The insoluble dietary fiber and soluble dietary fiber of the pumpkin sweet potato treated by enzyme were 4.17% and 2.07%, respectively. The spray-dried pumpkin sweet potato hydrolysates was manufactured via spray-drying with different forming agents: i.e., pectin 0.1%, 0.5%, 1%, and 2.0%. The moisture contents and total starches of the spray-dried powders were approximately 1.68-2.46 and 45.32-46.51%, respectively. The color of the L and a value decreased, and that of the b and ΔE value increased. The particle size and outer topology of the spray-dried powders were 37.17-42.32 μm, and its shape was generally globular. The water absorption index of the spray-dried powder (1.74-1.91) was lower than that of the freeze-dried powder (2.15). The water solubility index of the spray-dried powder, 80.75-87.61%, was higher than that of the freeze-dried powder (70.47%). The adhesion values of spray-dried powder to epithelial HT-29 cells were 2.66-6.18% of the initial cell counts, whereas freeze-dried powder showed lower adhesive ability (1.79%). The in vitro human digestibility in the spray-dried powder was 70.09% which is very effective in digestion.

Biodegradation of petroleum hydrocarbon functional groups in contaminated water using selected protozoan isolates

Diverse microorganisms are responsible for the biodegradation of various petroleum hydrocarbon chains which are found in both the environment and several existing water sources. Most of these microorganisms are both prokaryotic and eukaryotic in nature. The aim of this study was to investigate the degradation of petroleum hydrocarbon functional groups in contaminated water using individual protozoan species (Aspidisca sp., Trachelophyllum sp. and Peranema sp.) as well as a consortium of these three protozoan species. Chemical oxygen demand (COD) and dissolved oxygen (DO) were analysed using standard methods. The concentration of petroleum oil was determined in triplicate before and after the inoculation of oily wastewaters with the abovementioned protozoan species using the partition-gravimetric method. Various functional groups that had been biodegraded by the protozoan isolates were analysed using thin-layer chromatography (TLC) and GC×GC-FID. Results showed an increase in protozoan biomass from an initial cell count of 1.0×103cells/mL to a final cell count of 2.4×103cells/mL for the consortium of isolates, unlike a minimal biomass increases observed with individual isolates over the study period. The average uptake of DO was found to be low at 0.50mg/L from an initial concentration of 6.46mg/L to the final concentration of 6.00mg/L; however, the concentration of COD released by the isolates was found to be in an average range of ≤2300mg/L. The consortium of three isolates was able to biodegrade ≥80% of the hydrocarbon fractions, and aliphatic groups such as short-chain alkanes, alcohol/ester compounds and partially aromatic hydrocarbons were biodegraded, while the individual isolates were able to biodegrade an average of 72% of the amount of petroleum hydrocarbon fractions during the study.

Functional probiotic products based on fermented buckwheat with Lactobacillus rhamnosus

The fact that most probiotic products are milk based, naturally evokes the idea to develop other than dairy formulas. Therefore, the buckwheat mashes were fermented with lactic acid bacteria (LAB) of the Fresco DVS 1010 culture and probiotic strain Lactobacillus rhamnosus GG. The effect of single and co-culture fermentation was studied at 37 ± 0.5 °C for 8 h (5% CO2), afterwards with storage period for 14 days (6 ± 0.5 °C). Fresco culture showed good growth in all prepared mashes (specific growth rates ranged from 0.66 to 1.47 h−1), with higher counts in case of co-culture fermentation (8.8–44.9%). Probiotic strain Lb. rhamnosus GG during co-culture fermentation was able to grow from initial cell counts 5.9–6.7 log CFU ml−1 to final 8.4–9.0 log CFU ml−1 and levels higher than 6 log CFU ml−1 remained during cold storage. However, when LAB were in co-culture, 8 h was required to reach a pH value of 4.5 minimizing outgrowth of the contaminating microbiota. Based on the overall acceptance of final products, in case of single fermentation 14 days storage period gave the most acceptable products with points ranging from 3.3 to 3.7, while co-culture fermented products were the most acceptable after 7 days of storage (2.4–3.4).

Tenacity of Bacillus cereus and Staphylococcus aureus in dried spices and herbs

Numerous studies examined the antimicrobial effects of spice and herb extracts, whereas little is known about the effects of dry condiments on the survival of microorganisms. This study investigated the impact of dried condiments on the survival of Bacillus cereus and B. thuringiensis spores as well as Staphylococcus aureus cells. In addition, the survival variability between different strains was evaluated. Condiments (allspice, basil, cinnamon, nutmeg, oregano, paprika, parsley and pepper) were artificially contaminated by a dry spiking method using sand as carrier matrix and as control. The results show that counts of B. cereus and B. thuringiensis spores (initial spore count 5.6 ± 0.2 log10 cfu/g and 6.7 ± 0.1 log10 cfu/g, respectively) did not decline significantly in all condiments over a period of 50 weeks. In contrast, in some of the spiked test materials, cell counts of S. aureus (initial cell count 8.1 ± 0.5 log10 cfu/g) were reduced below the detection limit of 10 cfu/g within 10 weeks of storage. D values for S. aureus ranged between 5 and 31 days depending on the strain, condiment and initial contamination level. In conclusion, dried condiments may not affect the survival of spores but can significantly affect the survival of non-spore forming bacteria. As strain variability can occur, tenacity studies should be conducted including a variety of strains.

Endothelial Cell Count in Eye Bank Corneal Grafts: Impact of Death Cause and Donor Diseases

ABSTRACTPurpose: The purpose of this study is to analyze the impact of death causes and documented donor diseases on initial endothelial cell counts (after retrieval) and the development of corneal graft endothelia during organ culture. Methods: The retrospective statistic analyses was conducted on a data set of 10,185 human corneas prepared at the Hamburg Eye Bank. Results: Although we observed that death by gunshot trauma or alcoholism seems to be associated with marginally higher endothelium cell counts (independently from donor age), we could prove that only donor age is a relevant predictive parameter for the initial cell-density of the endothelium and its development in vitro. Conclusion: We conclude that an extension of prospective quality parameters for donor selection additional to donor age (such as individual causes of death) is not necessary.

Impact of spiking techniques on the survival of Staphylococcus aureus in artificially contaminated condiments

Tenacity studies are of relevance for microbial food safety risk assessment. Investigations on the survival of microorganisms in food matrices often require an artificial contamination step prior to microbial testing. Artificial contamination of low-moisture foods, like dried culinary herbs and spices, is challenging. In order to investigate the effect of several spiking techniques on the survival of Staphylococcus (S.) aureus in condiments, dried and ground paprika, pepper, and oregano were spiked with S. aureus using four different techniques: (1) aqueous bacteria suspensions were air-dried directly on the condiments; (2) aqueous bacteria suspensions were air-dried on sand, and then added to the condiments; (3) aqueous bacteria suspensions, or (4) bacteria suspended in liquid nutrient media with lyoprotectant were freeze-dried, crushed to powder, and added to the condiments. For each technique-condiment combination, and point in time, three distinct spiked samples (in three flasks) were prepared and stored in a dark environment at 24 ± 2 °C for up to 25 weeks. Cell counts directly after spiking and over storage time were analyzed. For the latter approach, mathematic models were used to fit the survival curve of each technique-condiment combination and to estimate their D-values. Results indicated that the reduction of the initial S. aureus cell counts depended on the applied spiking techniques and the used matrix. For instance, the initial cell counts in oregano were reduced by 0.8 ± 0.1 log10 cfu/g with technique 2 and by 6.1 ± 0.5 log10 cfu/g with technique 1. Technique 1 at the same time reduced the initial cell counts in paprika by only 0.4 ± 0.1 log10 cfu/g. The calculated D-values ranged from 1.3 ± 0.1 days (technique 3) to 120.1 ± 33.4 days (technique 4). The lowest as well as the highest D-value were observed for paprika samples, which emphasized the effect of the spiking technique. Thus, the behavior of bacteria can be strongly influenced by the selection of the spiking technique used for tenacity studies. Therefore, at least two spiking techniques should be considered, if the survival of microorganisms in low moisture food has to be investigated. In favor of high cell counts after drying, techniques 2 and 4 should be considered, as they performed best in our study.

Photo-inactivation of Escherichia coli and Enterococcus hirae using methylene blue and sodium anthraquinone-2-sulphonate: effect of process parameters.

In this study, effect of different parameters, viz. concentration of photosensitizer (PS), pH of the bacterial cell suspension and initial cell count, on photo-inactivation of Escherichia coli and Enterococcus hirae bacteria using methylene blue (MB) and sodium anthraquinone-2-sulphonate (SAQS) was investigated employing the statistically valid full factorial design of experiments. The inactivation efficiency of E. hirae using MB ranges between 10.81 and 48.55 %, whereas in the case of E. coli it ranges between 10.41 and 46.44 %. Using SAQS, the inactivation efficiency of E. hirae was within 5.26–39.03 %, and in the case of E. coli it varied in the range 4.65–37.66 %. Statistical analysis of the photo-inactivation results in the form of analysis of variance (ANOVA) and student ‘t’ test revealed significant individual effect of these process parameters. In addition, an increase in dark incubation period with MB or SAQS resulted in enhanced photo-inactivation efficiency against both the microorganisms. Reactive oxygen species measurement and analysis of lipid peroxidation and protein carbonyl index helped in a better understanding of the photo-inactivation mechanism.

A piecewise model of virus-immune system with two thresholds

The combined antiretroviral therapy with interleukin (IL)-2 treatment may not be enough to preclude exceptionally high growth of HIV virus nor rebuilt the HIV-specific CD4 or CD8 T-cell proliferative immune response for management of HIV infected patients. Whether extra inclusion of immune therapy can induce the HIV-specific immune response and control HIV replication remains challenging. Here a piecewise virus-immune model with two thresholds is proposed to represent the HIV-1 RNA and effector cell-guided therapy strategies. We first analyze the dynamics of the virus-immune system with effector cell-guided immune therapy only and prove that there exists a critical level of the intensity of immune therapy determining whether the HIV-1 RAN virus loads can be controlled below a relative low level. Our analysis of the global dynamics of the proposed model shows that the pseudo-equilibrium can be globally stable or locally bistable with order 1 periodic solution or bistable with the virus-free periodic solution under various appropriate conditions. This indicates that HIV viral loads can either be eradicated or stabilize at a previously given level or go to infinity (corresponding to the effector cells oscillating), depending on the threshold levels and the initial HIV virus loads and effector cell counts. Comparing with the single threshold therapy strategy we obtain that with two thresholds therapy strategies either virus can be eradicated or the controllable region, where HIV viral loads can be maintained below a certain value, can be enlarged.

&lt;italic&gt;Escherichia Coli&lt;/italic&gt; Inactivation in Water Using Pulsed Discharge

In this paper, a new multiple-needle-cylinder reactor is designed to study the Escherichia coli inactivation with an unstable pulsed arc discharge in water and investigate the influence of several parameters for promoting its industrial application. The results show that the inactivation rate is mainly determined by energy density. About three orders reduction can be obtained with an initial cell count of $3\times 10^{5}$ CFU/mL and energy density around 2 J/mL. The inactivation rate can almost reach 100% when the energy density is around 8 J/mL. For reaching the maximum inactivation rate of this system, the parameters including initial water conductivity below $50~\mu \text{S}$ /cm, initial cell count over $10^{4}$ CFU/mL, and bubbling flow rate around 3.5 L/min are recommended. The water conductivity, pH value, and concentrations of hydrogen peroxide and ozone are also measured during the treatment to evaluate their influence. The transmission electron microscopy images of E. coli indicate that the intracellular morphology changes after plasma treatment.

Cord blood collection and processing with hydroxyethyl starch or non–hydroxyethyl starch

BackgroundCollection and processing characteristics influencing quality of cord blood (CB) units play an essential role to cord blood banks (CBBs). At many CBBs, volume reduction is performed using hydroxyethyl starch (HES) and the Sepax (Biosafe) automated cell processing system. Due to the withdrawal of HES from the European market, a validation of the nonHES protocol was performed. MethodsThis partially retrospective study identified CB characteristics such as gestational age and CB volume/cell count correlated with higher quality. For the nonHES validation, CB was analyzed for total nucleated cell (TNC), mononuclear cell (MNC) recovery, hematocrit (HCT) and colony-forming units (CFUs). Viabilities of CD34+ and CD45+ cells were determined by 7-aminoactinomycin D (7-AAD) and AnnexinV (AnnV) staining and compared for 21 mL and 42 mL buffy coat (BC) samples applying the HES/nonHES protocol. ResultsFactors affecting the potency of CB transplants were the gestational age and the volume reduction to a defined BC volume. High initial cell counts and CB volumes correlated negatively with post-processing TNC recovery for lower BC volumes. Post-processing HES and nonHES results were comparable, but nonHES revealed a significantly lower post-thaw recovery of viable CD34+ cells measured by 7-AAD/AnnV (21 mL: 45.4 ± 16.4%; 42 mL: 67.3 ± 14.5%) as compared with HES (21 mL: 72.7 ± 14.4%, P = 0.0164; 42 mL: 83.4 ± 14.7%, P = 0.0203). DiscussionDue to the lower post-thaw CD34+ cell viability (AnnV−/7-AAD−) for nonHES samples, the use of HES is recommended, ideally combined with a high BC volume. The post-processing HCT has no statistically significant impact on the post-thaw CD34+ cell viability (AnnV−/7-AAD−).