Nineteen of the pharmacists worked in a variety of different phar

Nineteen of the pharmacists worked in a variety of different pharmacies,

both independents and multiples. Six worked regularly in one or two pharmacies. Verbatim transcripts underwent directed content analysis using NVivo software. Ethical approval was obtained from the University of Central Lancashire Research Ethics Committee. Locums reported a rapid process of assessing staff competence and also identified the possible safety risks in attempting to change usual practice in the pharmacy. Resistance of staff to locum authority was described. Locums also reported a lack of support from employers in managing difficulties with staff, with threats to future employment if issues were raised. Assessing staff competence and work processes was seen as important for safety: “you’ve got 17-AAG to be able to pick up very quickly how the staff in that place work, to allow them to do their job as they feel comfortable so they don’t make mistakes” (FG2 male, over

40). Change in processes was identified as a possible risk: “it would be very dangerous to get the staff to change for one day, so you don’t, you work with it” (FG1 male, over 40). Passive undermining of locums by staff was noted: “[staff] say come back when the regular pharmacist is in even though you’re here and you can help” (FG1 female, under 40) and also more active, even aggressive behaviour: “[staff] were banging on the [consultation room] door and they were shouting at me, ‘come on you’ve got prescriptions out here, come on hurry up’ ” (FG5 female, under 40). Locums perceived a lack of support buy AZD4547 from employers for these issues: “I know for 100 percent… they will always, always favour their own

staff over you triclocarban as a locum because they don’t need you…they’ll keep their own staff happy so that the staff will run the shop for them” (FG3 male, under 40). Further employment was also potentially at risk: “the company didn’t do anything they just said you’ve got to put up with her or don’t come back” (FG2 male, under 40). This paper describes a sometimes difficult working environment for locum community pharmacists, involving them assessing and managing risk to patients during the working interactions with staff. This can present a challenge to locum professional autonomy, where locums may be in conflict with staff over patient care issues. This challenge is compounded by risks to future employment when issues are raised with company management. The impact on patient care of pharmacies run entirely on varied locum staff is worthy of further study. 1. Shann, P. and Hassell, K. 2004, An exploration of the diversity and complexity of the pharmacy locum workforce, Royal Pharmaceutical Society of Great Britain, London. A. Tonnaa, A. Weidmanna, R. Laingb, I. Tonnab, G. McCartneyb, D.

AIDS 2009; 23: 875–885 73 Silverberg MJ, Chao C, Leyden WA et al

AIDS 2009; 23: 875–885. 73 Silverberg MJ, Chao C, Leyden WA et al. HIV infection, immunodeficiency, viral replication, and the risk of cancer. Cancer Epidemiol Biomarkers Prev 2011; Apoptosis inhibitor 20: 2551–2559. 74 Silverberg MJ, Chao C, Leyden WA et al. HIV infection status, immunodeficiency, and the incidence of non-melanoma skin cancer. J Natl Cancer Inst 2013; 105: 350–360. 75 Newnham A, Harris J, Evans HS et al. The risk of cancer in HIV-infected people in southeast England: a cohort study. Br J Cancer 2005; 92: 194–200. 76 Marsden JR, Newton-Bishop JA, Burrows L et al.; British Association of Dermatologists Clinical Standards Unit. Revised UK guidelines for the management of cutaneous

melanoma 2010. Br J Dermatol 2010; 163: 238–256. 77 van Leeuwen MT, Vajdic CM, Middleton MG et al. Continuing declines in some but not all HIV-associated cancers in Australia

after widespread use of antiretroviral therapy. buy Z-VAD-FMK AIDS 2009; 23: 2183–2190. 78 de Berker D, McGregor JM, Hughes BR; British Association of Dermatologists Therapy Guidelines and Audit Subcommittee. Guidelines for the management of actinic keratoses. Br J Dermatol 2007; 156: 222–230. 79 de Boer WA, Danner SA. HIV infection and squamous cell carcinoma of sun exposed skin. AIDS 1990; 4: 91. 80 Kreuter A, Weiland U, Brockmeyer NH, German Network of Competence HIV/AIDS. Genital human papillomavirus-associated (pre-) malignant skin diseases drastically increase in the era of highly active antiretroviral therapy for HIV infection. J Am Acad Dermatol 2006; 55: 1116–1117. 81 Maurer TA, Christian KV, Kerschmann RL et al. Cutaneous squamous cell carcinoma in human immunodeficiency virus-infected patients. A study of epidemiologic risk factors, human papillomavirus, and p53 expression. Arch Dermatol Exoribonuclease 1997; 133: 577–583. 82 Kreuter

A, Brockmeyer NH, Pfister H et al. Competence Network HIV/AIDS. Human papillomavirus type 26-associated periungual squamous cell carcinoma in situ in a HIV-infected patient with concomitant penile and anal intraepithelial neoplasia. J Am Acad Dermatol 2005; 53: 737–739. 83 Fearfield LA, Nelson M, Francis N, Bunker CB. Cutaneous squamous cell carcinoma with zosteriform metastases in a human immunodeficiency virus-infected patient. Br J Dermatol 2000; 142: 573–574. 84 Neves-Motta R, Ferry FR, Basilio-de-Oliveira CA et al. Highly aggressive squamous cell carcinoma in an HIV-infected patient. Rev Soc Bras Med Trop 2004; 37: 496–498. 85 Nguyen P, Vin-Christian K, Ming ME, Berger T. Aggressive squamous cell carcinomas in persons infected with the human immunodeficiency virus. Arch Dermatol 2002; 138: 758–763. 86 Hausauer AK, Maurer T, Leslie KS et al. Recurrence after treatment of cutaneous basal cell and squamous cell carcinomas in patients infected with human immunodeficiency virus. JAMA Dermatol 2013; 149: 239–241. 87 Kagen MH, Hirsch RJ, Chu P et al.

Ninety-eight nanograms of the 5′ HEX-labelled TN or BN suicide su

Ninety-eight nanograms of the 5′ HEX-labelled TN or BN suicide substrates were incubated in 20 μL-reaction volumes containing TDMNG buffer (50 mM Tris pH 7.5, 5 mM DTT, 75 mM MgCl2, 25 mM NaCl and 25% glycerol) in the presence of 1.5 mM MBP-XerS each in the presence of 1 μg poly dI-dC. After a 60 min incubation at 37 °C, reactions were stopped with 5 μL

of 2% SDS and 5 μL of Orange loading dye (NEB), incubated at 100 °C for 10 min and then electrophoresed in a 6% TBE gel in the presence of 0.1% SDS, and scanned with a EPZ5676 mouse Typhoon imager. The difSL cleavage site was determined using 98 ng of the 3′ FITC-labelled TN or BN suicide substrate and was incubated in 20 μL of TDMNG buffer in the presence of variable concentrations of MBP-XerS in the presence of 1 μg Entinostat chemical structure poly dI-dC. After a four-hour incubation at 37 °C, reactions were stopped with 20 μL of formamide, incubated at 75 °C for 2 min and then electrophoresed in

a 20% polyacrylamide TBE gel with 6 M urea, and scanned with a Typhoon imager. Molecular weight ladders were prepared by chemical degradation of the 3′ FITC-labelled oligonucleotides following the G+A chemical sequencing protocol (Bencini et al., 1984). Under the conditions used, cleavage was observed at each nucleotide position, generating a ladder of fragments differing by a single nucleotide. The thermosensitive plasmid pBEA756 was used to inactivate the xerS gene of S. suis (Fittipaldi et al., 2007). An internal sequence of the

xerS gene was amplified by PCR and cloned into the EcoRI site of pBEA756, forming the plasmid pBEAXerCint. Plasmids were then electroporated into S. suis from (prepared according to Pulliainen et al., 2003) using a Bio-Rad gene pulser using 0.2-cm cuvettes at 2.5 kV. Immediately after the pulse, 1 mL of cold THY medium supplemented with 0.3 M sucrose was added, and the samples were incubated at 28 °C for 3 h, and spread on a THA plate containing 1% yeast, 400 μg mL−1 kanamycin and incubated at 28 °C. The resulting transformants were then grown in THY broth overnight with kanamycin selection at 28 °C. Aliquots of overnight cultures were spread on selective THA plates and incubated at 37 °C to inactivate the gram-positive origin. Cells which remained kanamycin resistant, presumably had integrated the plasmid into the chromosome by homologous recombination at the xerS locus, inactivating the gene. This was confirmed by Southern blot analysis, using genomic DNA prepared from kanamycin-resistant cultures using the DNeasy tissue kit. Complementation of the xer− phenotype was observed after re-introducing a cloned xerS gene with its promoter into pGhost9, and electroporating the construct into xerS mutant cells.

An early assessment of the social circumstances of a newly diagno

An early assessment of the social circumstances of a newly diagnosed HIV-positive woman is important. Patients who initially refuse interventions or default from follow-up need to be identified and actively followed-up. Support by trained peer-support workers is a valuable component of the management of HIV-positive pregnant women. Many newly diagnosed HIV-positive pregnant women are initially selleck chemicals reluctant to engage with peer support; however, the great majority of women who do engage with it find that it becomes one of the most highly valued of all the interventions that they undertake [314]. The importance of informing appropriate healthcare

workers should be emphasized. This includes midwives, general practitioners, health visitors and paediatricians. The process of in-patient care should be explained clearly, so that the women can be helped to inform ward staff explicitly about levels of disclosure to visitors. Depending on the setting, levels of disclosure of newly diagnosed pregnant women about their HIV

status vary, and there are cultural factors that influence the patterns of self-disclosure to partners and other social network members [313],[315]. Disclosure should be encouraged in all cases but may be viewed as a process that may take some time [316, 317]. There are find more situations where a newly diagnosed HIV-positive woman refuses to disclose to a current sexual partner, or appears to want to delay disclosure indefinitely. This can give rise to very complex professional, ethical, moral and, potentially, legal situations. There is a conflict between the duty of confidentiality to the index patient and a duty to prevent Sclareol harm to others. Breaking confidentiality to inform a sexual partner of the index patient’s positive HIV status is sanctioned as a ‘last resort’ by the WHO [318] and General Medical Council [319]. However, it is not to be taken lightly as it could have the negative impact of deterring others from testing because of the fear of forced disclosure and loss of trust by patients in the confidential doctor–patient relationship.

Difficult disclosure cases should be managed by the MDT. It is important to accurately record discussions and disclosure strategy in difficult cases. Simultaneous partner testing during the original antenatal HIV test should be encouraged wherever possible, as couples will frequently choose to receive their HIV test results together, providing simultaneous disclosure. Reassurance about confidentiality is extremely important, especially regarding family members and friends who may not know the diagnosis but are intimately involved with the pregnancy. Women from communities with high levels of HIV awareness may be concerned about HIV ‘disclosure-by-association’ when discussing certain interventions, including taking medication during pregnancy, having a CS, and avoiding breastfeeding.

An early assessment of the social circumstances of a newly diagno

An early assessment of the social circumstances of a newly diagnosed HIV-positive woman is important. Patients who initially refuse interventions or default from follow-up need to be identified and actively followed-up. Support by trained peer-support workers is a valuable component of the management of HIV-positive pregnant women. Many newly diagnosed HIV-positive pregnant women are initially PLX4032 solubility dmso reluctant to engage with peer support; however, the great majority of women who do engage with it find that it becomes one of the most highly valued of all the interventions that they undertake [314]. The importance of informing appropriate healthcare

workers should be emphasized. This includes midwives, general practitioners, health visitors and paediatricians. The process of in-patient care should be explained clearly, so that the women can be helped to inform ward staff explicitly about levels of disclosure to visitors. Depending on the setting, levels of disclosure of newly diagnosed pregnant women about their HIV

status vary, and there are cultural factors that influence the patterns of self-disclosure to partners and other social network members [313],[315]. Disclosure should be encouraged in all cases but may be viewed as a process that may take some time [316, 317]. There are GSK458 ic50 situations where a newly diagnosed HIV-positive woman refuses to disclose to a current sexual partner, or appears to want to delay disclosure indefinitely. This can give rise to very complex professional, ethical, moral and, potentially, legal situations. There is a conflict between the duty of confidentiality to the index patient and a duty to prevent Fenbendazole harm to others. Breaking confidentiality to inform a sexual partner of the index patient’s positive HIV status is sanctioned as a ‘last resort’ by the WHO [318] and General Medical Council [319]. However, it is not to be taken lightly as it could have the negative impact of deterring others from testing because of the fear of forced disclosure and loss of trust by patients in the confidential doctor–patient relationship.

Difficult disclosure cases should be managed by the MDT. It is important to accurately record discussions and disclosure strategy in difficult cases. Simultaneous partner testing during the original antenatal HIV test should be encouraged wherever possible, as couples will frequently choose to receive their HIV test results together, providing simultaneous disclosure. Reassurance about confidentiality is extremely important, especially regarding family members and friends who may not know the diagnosis but are intimately involved with the pregnancy. Women from communities with high levels of HIV awareness may be concerned about HIV ‘disclosure-by-association’ when discussing certain interventions, including taking medication during pregnancy, having a CS, and avoiding breastfeeding.

, 1999) Serum opacification activity has been observed in other

, 1999). Serum opacification activity has been observed in other streptococci, such as Streptococcus pyogenes and S. suis (Ward & Rudd, 1938; Baums et al., 2006). Serum opacity factor (SOF) is a bifunctional protein consisting of highly

conserved C-terminal repetitive sequences, and the N-terminal serum opacification domain (Rakonjac et al., 1995; Courtney et al., 2002). SOF caused S. pyogenes to invade and adhere to cells, and was demonstrated to be a virulence determinant of S. pyogenes using a murine infection model (Timmer et al., 2006; Gillen et al., 2008). The repetitive ABT-199 research buy C-terminal fibronectin-binding domains and high similarity in part of the N-terminal domain between serum opacity genes were observed in FnBA of S. dysgalactiae strain S2 and several SOFs from S. pyogenes strains (Courtney et al., 1999; Katerov et al., 2000). Although many variable sequences of sof genes exist in S. pyogenes, only a few genes coding SOF were reported to exist in S. dysgalactiae isolates from mammals. GCSD isolated from fish also possesses serum opacification activity. However, the gene encoding activity has been not identified. The aim of this study was to identify the sof gene, named sof-FD, and to determine its distribution in GCSD isolated from farmed fish. The designed oligonucleotides targeting

sof-FD were applied to a PCR assay to discriminate between fish GCSD and mammalian S. dysgalactiae. A total of 316 GCSD strains, isolated from farmed fish (amberjack, 276; yellowtail, 40) between I-BET-762 datasheet 2002 and 2008 in Japan, were used to detect SOF. Streptococcus

dysgalactiae isolated from pigs (n = 17) diagnosed with endocarditis in the Kumamoto Prefectural Meat Inspection Office was used as the source of mammalian isolates. Lancefield streptococcal grouping (Lancefield, 1933) was performed for these isolates using a Pastorex Strep Ribonuclease T1 test (Bio-Rad, Marnes-la-Coquette, France). The fish and mammalian isolates were identified using a PCR assay targeting the 16S–23S rRNA spacer region (Forsman et al., 1997; Hassan et al., 2003). Streptococcus dysgalactiae ssp. dysgalactiae ATCC 43078 and S. dysgalactiae ssp. equisimilis ATCC 35666 were used as reference strains. All the isolates were cultured on Todd-Hewitt (TH) agar (Difco, Sparks, MD) at 37 °C for 24 h. Serum opacification activity was detected using the microtitre plate method (Johnson & Kaplan, 1988) with minor modifications. Bacterial strains were cultured in TH broth at 37 °C for 24 h. The supernatants or 0.5% (sodium dodecyl sulfate) SDS extracts of bacterial cells were filtered using a 0.45-μm filter (Sartorius Stedim Japan K. K., Japan). Fish serum was obtained from healthy amberjacks (average weight 1250 g, n = 15). Briefly, fish were anaesthetized using FA-100 (Tanabe Pharma, Osaka, Japan), then bled from the caudal peduncle. After clotting of blood, the serum was separated by centrifugation for 20 min at 5000 g.

1 and Kv21) and one G-protein-gated

inwardly rectifying

1 and Kv2.1) and one G-protein-gated

inwardly rectifying (Kir3.2) K+ channel subunits on hippocampal CA1 pyramidal cells (PCs). Freeze-fracture replica immunogold labelling was employed to determine the relative densities of these K+ channel subunits in 18 axo-somato-dendritic compartments. Significant densities of the Kv1.1 subunit were detected on axon initial segments (AISs) and axon terminals, with an approximately eight-fold lower density in the latter compartment. The Kv2.1 subunit was found in somatic, proximal dendritic and AIS plasma membranes at approximately Selleckchem DAPT the same densities. This subunit has a non-uniform plasma membrane distribution; Kv2.1 clusters are frequently adjacent to, but never overlap with, GABAergic synapses. A quasi-linear increase in the Kir3.2 subunit density along the dendrites of PCs was detected, showing no significant difference between apical dendritic shafts, oblique dendrites or dendritic

spines at the same distance from the soma. Our results demonstrate that each subunit has a unique cell-surface distribution pattern, and predict their differential involvement in synaptic integration and output generation at distinct subcellular compartments. “
“Nerve growth factor (NGF) signaling is important in the development and functional maintenance of nociceptors, but it also plays a central role in initiating and sustaining heat and mechanical hyperalgesia following inflammation. NGF signaling in pain has traditionally been thought of as primarily engaging the classic high-affinity receptor tyrosine kinase receptor TrkA to initiate sensitization events. However, the discovery selleck that secreted proforms of nerve NGF have biological functions distinct from the processed mature factors raised the possibility Rebamipide that these proneurotrophins (proNTs) may have distinct function in painful conditions. ProNTs engage a novel receptor system that is distinct from that of mature neurotrophins, consisting of sortilin, a type I membrane protein belonging to the VPS10p family, and its co-receptor, the classic

low-affinity neurotrophin receptor p75NTR. Here, we review how this new receptor system may itself function with or independently of the classic TrkA system in regulating inflammatory or neuropathic pain. “
“A growing body of evidence suggests that gonadal steroids such as estradiol (E2) alter neural responses not only in brain regions associated with reproductive behavior but also in sensory areas. Because catecholamine systems are involved in sensory processing and selective attention, and because they are sensitive to E2 in many species, they may mediate the neural effects of E2 in sensory areas. Here, we tested the effects of E2 on catecholaminergic innervation, synthesis and activity in the auditory system of white-throated sparrows, a seasonally breeding songbird in which E2 promotes selective auditory responses to song.

Patients starting d4T on the lower dose who gained weight to abov

Patients starting d4T on the lower dose who gained weight to above 60 kg were changed to the higher dose. As per clinical guidelines, lactate

measurements are requested in symptomatic patients only. The existing case series from which the cases were drawn describes the clinical management of SHLA in this setting, as well as the referral rates, characteristics and outcomes of referred patients with SHLA [18]. In the published case series the referral rate was 17.5 [95% confidence interval (CI) 13.7–21.9] per 1000 patient-years for SHLA, and 12.1 (95% CI 9.2–16.1) per 1000 patient-years for lactic acidosis (53 of the 75 cases in the full series were acidotic, and the median lactate value was 7.6 mmol/L [interquartile range Rucaparib in vitro (IQR) 5.9–9.8]). Acute mortality was 16% for SHLA and 21% for lactic acidosis. A matched case–control study was conducted using incidence density sampling and builds on the case series reported by Stead et al. [18] This case–control study was nested within the larger cohort of ART patients attending public sector ART services in the province [19]. All patients with lactate ≥5 mmol/L referred to GF Jooste Hospital between 1 August 2003 and 15 November 2005 were considered. Potential cases with alternative aetiology to explain a raised lactate, including hepatitis, severe dehydration and sepsis, were excluded from the study. The resulting sample size of 5-FU concentration 71 cases provided 80% power to detect a 3-fold

difference in the risk of SHLA for women compared with men and for weight above 70 kg, assuming two controls for each case. These effect sizes were well within those described in a smaller cohort study in the same setting [17]. Two systematically selected controls were matched to their respective cases by primary health care facility and duration on ART. Matching by facility

was necessary because of the nature of the information system, Smoothened while matching by duration was by design, to avoid over-representing patients who had recently started ART. Controls were considered eligible if they were still in care at the facility at the time of the SHLA diagnosis of their matched case. Selected controls had to be treatment-naïve and not have a determined lactate ≥5 mmol/L between ART initiation and the SHLA presentation date of their matched case. Nonreplacement selection was used; however, because of the small numbers initiating therapy per facility at the beginning of the national ART roll-out, four controls were selected twice. All baseline and longitudinal data were collected retrospectively from each participant’s primary care folder. Follow-up data were collected from ART initiation to either case presentation for the cases or the date of presentation for each control’s matched case. Variables at baseline included demographic information, WHO stage-defining illnesses, concomitant chronic medical conditions, tuberculosis history, baseline laboratory results and clinical assessment details.

Patients starting d4T on the lower dose who gained weight to abov

Patients starting d4T on the lower dose who gained weight to above 60 kg were changed to the higher dose. As per clinical guidelines, lactate

measurements are requested in symptomatic patients only. The existing case series from which the cases were drawn describes the clinical management of SHLA in this setting, as well as the referral rates, characteristics and outcomes of referred patients with SHLA [18]. In the published case series the referral rate was 17.5 [95% confidence interval (CI) 13.7–21.9] per 1000 patient-years for SHLA, and 12.1 (95% CI 9.2–16.1) per 1000 patient-years for lactic acidosis (53 of the 75 cases in the full series were acidotic, and the median lactate value was 7.6 mmol/L [interquartile range Epacadostat chemical structure (IQR) 5.9–9.8]). Acute mortality was 16% for SHLA and 21% for lactic acidosis. A matched case–control study was conducted using incidence density sampling and builds on the case series reported by Stead et al. [18] This case–control study was nested within the larger cohort of ART patients attending public sector ART services in the province [19]. All patients with lactate ≥5 mmol/L referred to GF Jooste Hospital between 1 August 2003 and 15 November 2005 were considered. Potential cases with alternative aetiology to explain a raised lactate, including hepatitis, severe dehydration and sepsis, were excluded from the study. The resulting sample size of Selleckchem PD0332991 71 cases provided 80% power to detect a 3-fold

difference in the risk of SHLA for women compared with men and for weight above 70 kg, assuming two controls for each case. These effect sizes were well within those described in a smaller cohort study in the same setting [17]. Two systematically selected controls were matched to their respective cases by primary health care facility and duration on ART. Matching by facility

was necessary because of the nature of the information system, MYO10 while matching by duration was by design, to avoid over-representing patients who had recently started ART. Controls were considered eligible if they were still in care at the facility at the time of the SHLA diagnosis of their matched case. Selected controls had to be treatment-naïve and not have a determined lactate ≥5 mmol/L between ART initiation and the SHLA presentation date of their matched case. Nonreplacement selection was used; however, because of the small numbers initiating therapy per facility at the beginning of the national ART roll-out, four controls were selected twice. All baseline and longitudinal data were collected retrospectively from each participant’s primary care folder. Follow-up data were collected from ART initiation to either case presentation for the cases or the date of presentation for each control’s matched case. Variables at baseline included demographic information, WHO stage-defining illnesses, concomitant chronic medical conditions, tuberculosis history, baseline laboratory results and clinical assessment details.

, 2007; Zhou et al, 2009; da Miguel et al, 2010),

such

, 2007; Zhou et al., 2009; da Miguel et al., 2010),

such methods may provide an inaccurate description of the total microbial structure in that they reveal only dominant populations, which may not necessarily Ipilimumab datasheet play important roles in overall community dynamics. Lacticin 3147 is a potent, two-peptide broad spectrum lantibiotic (class I bacteriocin or antimicrobial peptide) produced by Lactococcus lactis DPC3147 (Fig. 1; Ryan et al., 1996; Martin et al., 2004; Lawton et al., 2007). First isolated from an Irish kefir grain in 1996, it is perhaps one of the most extensively studied bacteriocins and has been shown to inhibit such clinically relevant pathogens as Clostridium difficile, GSK126 price methicillin-resistant Staphylococcus aureus and vancomycin-resistant enterococci (Rea et al., 2007; Piper et al., 2009). Although the microbial composition of kefir grains has been well documented (Rea et al., 1996; Ninane et al., 2007;

Zhou et al., 2009), to our knowledge, there have been no reports on the characterization of the microbiota of a kefir grain from which bacteriocin-producing strains have been isolated. In recent years, the field of microbial ecology has been revolutionized by the development and application of high-throughput DNA sequencing technologies, such as that facilitated by the 454 GS-FLX platform (Roche Diagnostics Ltd, West Sussex, UK; Keijser et al., 2008; Urich et al., 2008; McLellan et al., 2009), which allows for a more complete view of overall community composition without the bias typically associated with cloning or cultivation. Here, we use high-throughput sequencing of 16S rRNA gene amplicons to characterize the bacterial composition of the original Irish kefir from which L. lactis DPC3147 was initially isolated. The kefir grain starter used in this study was obtained from the Teagasc Food Research Centre (Fig. 1a; Teagasc, Fermoy, Ireland) kefir grain collection. The grain was cultured in sterile 10%

reconstituted skim milk at 21 °C for 24 h. The fermented Interleukin-3 receptor kefir milk was removed and the grain rinsed with sterile water to remove any clotted milk still adhered onto the grain surface. In order to monitor bacterial changes over the course of the kefir fermentation, kefir milk samples were enumerated for lactococci and lactobacilli; populations typically associated with the kefir community. Samples were first homogenized as 10-fold serial dilutions, further 10-fold serial dilutions were prepared and appropriate dilutions were spread plated onto M17 agar supplemented with 0.5% lactose (LM17; Difco Laboratories, Detroit, MI) for lactococci, and Lactobacillus selection agar (LBS; Difco) for lactobacilli populations. LM17 plates were incubated aerobically at 30 °C overnight and LBS plates were incubated anaerobically at 37 °C for 5 days.