Search terms used across databases.

<p>Search terms used across databases.</p

PRISMA Flow diagram outlining the systematic review process.

<p>PRISMA Flow diagram outlining the systematic review process.</p

Rates of sleep disturbance found in studies where sleep was measured or quantified within the sample.

<p>Studies where the whole sample had sleep disturbance or where the number of those experiencing sleep disturbance are not included in this table.</p

Information (i.e. aim, study design, primary focus of the study, measure of sleep used, sample, pertinent results relating to sleep) and quality scores for each study using an unvalidated measure of sleep included in the review.

<p>Information (i.e. aim, study design, primary focus of the study, measure of sleep used, sample, pertinent results relating to sleep) and quality scores for each study using an unvalidated measure of sleep included in the review.</p

Inclusion criteria for studies.

<p>Inclusion criteria for studies.</p

Information (i.e. aim, study design, primary focus of the study, measure of sleep used, sample, pertinent results relating to sleep) and quality scores for each study using a validated measure of sleep included in the review.

<p>Information (i.e. aim, study design, primary focus of the study, measure of sleep used, sample, pertinent results relating to sleep) and quality scores for each study using a validated measure of sleep included in the review.</p

IFNγ increases PRL immunoreactivity in the hair follicle, whilst TNFα decreases follicular PRL immunoreactivity.

<p>(A) PRL IR in control hair follicle outer root sheath keratinocytes (black arrows), (B) after INFγ treatment and (C) negative control. (D) PRLR IR is significantly increased by INFγ treatment (p = 0.044). Results were pooled from 3 ♀ subjects (Aged 44–68), 14–16 HFs per group in total. In contrast, TNFα 5 ng/ml significantly decreased PRL IR (E). PRL IR is shown in (F) control hair follicles and those treated with (G) TNFα 0.5 ng/ml and (H) TNFα 5 ng/ml. Results were pooled from 3 ♀ subjects (Aged 49–68) 18–23 HFs per group in total. Scale bars represent 50 µm.</p

STAT5 phosphorylation is increased by PRL treatment in serum-free skin organ culture.

<p>Treatment with PRL (400 ng/ml) in serum-free organ culture increased epidermal STAT5 phosphorylation in all 3 of the individuals investigated. 3 females aged 42–63 years as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0060819#pone-0060819-g001" target="_blank">Fig 1(J)</a> and 2(I). Arrows show phosphorylated STAT5 as detected by monoclonal rabbit antibody recognising STAT5a and b phosphorylated at Tyr 694.</p

TNFα significantly decreases PRLR immunoreactivity in the outer root sheath of hair follicles.

<p>(A) PRLR IR in the outer root sheath of control hair follicles (black arrows) was reduced in comparison PRLR IR after treatment with TNFα at concentrations of (B) 0.5 ng/ml and (C) 5 ng/ml. (D) Negative control. (E) Quantitative analysis confirms decreased PRLR IR after TNFα treatment (13–17 HFs per group in total). Results were pooled from same subjects described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0060819#pone-0060819-g004" target="_blank">Fig 4</a>. (F) Pooled results of PRLR steady state gene expression in two subjects (♀ aged 53 and 66 years) showed no significant effect of 50 ng TNFα treatment after 48 hours. Scale bars represent 50 µm.</p

Prolactin immunoreactivity is present in corporal skin and its gene transcription can increase during culture.

<p>Prolactin (PRL) immunoreactivity (IR) is demonstrated in (A) axillary skin from a 49 year old ♀, (B) axillary skin from a 28 year old ♀ and (C) thigh skin from a 57 year old ♀. The black arrows show that the IR was most prominent in the basal layer of the epidermis, in a cytoplasmic distribution, in non-scalp skin. (D) Omission of the primary antibody served as the negative control and (E) the hair follicle outer root sheath keratinocytes and sebaceous gland served as internal positive controls. (F) PRL gene expression was below the level of detectability at day 0, but became detectable by day 7 during serum free organ culture in breast skin (from a 42 year old ♀) and (G) abdominal skin (from 63 year old ♀). At the protein expression level, PRL IR also increased between (H) day 0 and (I) day 7. (J) High magnification of PRL IR. Quantitative measurement of PRL IR, using Image J software is shown in (K). Results pooled from 3 females aged 42–63 years. Increased protein expression correlated with increased gene expression in two subjects over the same time period. However, in one subject PRL gene expression was readily detectable at day 0, but decreased significantly during organ culture (data not shown). qRT-PCR results could not be normalized, combined and expressed as fold change from day 0, as no PRL gene expression was detected at day 0 in 2/3 subjects. All scale bars represent 50 µm.</p