J Clin Gynecol Obstet

Journal of Clinical Gynecology and Obstetrics, ISSN 1927-1271 print, 1927-128X online, Open Access

Article copyright, the authors; Journal compilation copyright, J Clin Gynecol Obstet and Elmer Press Inc

Journal website http://www.jcgo.org

Original Article

Volume 4, Number 3, September 2015, pages 237-250

Urogenital Biomolecular and Physical Measures in Pre- and Post-Menopausal Women

Figure 1. Incidence of subjective urogenital symptoms among test groups. Panelists were asked to rate specific urogenital symptoms, including genital skin dryness and itch, and vaginal dryness and itch on a four-point scale (none, slight, moderate, or considerable). Perceived difficulty having intercourse was rated on the five-point scale (none, slight, moderate, considerable, or unbearable). The proportion of individuals in each test group claiming some degree of symptoms is plotted. Pairwise comparisons were conducted using Fisher’s exact test. Significant differences between groups are indicated on the graph.

Figure 2. Skin temperature and pH among test groups. Skin temperature (a) and pH (b) were obtained from three different body sites; labia majora, labia minora and introitus. The upper thigh was uses as a control site for the skin temperature measurement. In addition, vaginal pH was measured using litmus paper against the middle to upper third of the lateral wall of the vagina. Pairwise comparisons were conducted using a mixed linear model. Significant differences between groups are indicated on the graph.

Figure 3. Relationship between histamine levels and subjective symptoms. The levels of histamine detected from individuals who claimed the presence of subjective symptoms was compared to that from individuals who did not claim to experience the symptoms. The entire test population was considered as a whole, regardless of group assignment. Pairwise comparisons were conducted using Fisher’s exact test. Significant differences (P ≤ 0.05) and trends (P ≤ 0.1) between groups are indicated on the graph.

Figure 4. Changes in histamine and histidine at different life stages. The histamine (a), histidine (b) and histidine/histamine ratios (c) at three anatomic sites (given in Table 2) are plotted for each group to illustrate the change from the premenstrual group to the post-menstrual groups. Decrease from pre-M levels for post-M non-HRT group are illustrated by the pink arrows and text boxes, and for Post-M HRT group by the green arrows and text boxes.

**Table 1.** Summary of Demographics and Baseline Vaginal Atrophy Scores
Parameter	Pre-menopausal group (pre-M) (N = 15)	Post-menopausal, non-HRT group (post-M non-HRT) (N = 15)	Post-menopausal with HRT group (post-M HRT) (N = 15)
NA: not applicable. ^aPost-M HRT group significantly different from pre-M and post-M non-HRT groups (P < 0.0001).
	Mean ± SD	Mean ± SD	Mean ± SD
Age (years)	33.0 ± 6.4	60.7 ± 3.6	60.5 ± 3.6
Height (inch)	64.7 ± 3.7	63.5 ± 2.8	63.9 ± 2.5
Weight (lb)	156.0 ± 22.8	149.2 ± 27.0	147.8 ± 23.7
BMI (%)	26.2 ± 3.3	25.9 ± 3.6	25.5 ± 4.3
Years since last period (years)	NA	15.8 ± 9.3	14.2 ± 8.2
Average time on HRT	NA	NA	5 years 2 months

	Number (%)	Number (%)	Number (%)
Ethnicity
African-American	6 (40%)	2 (13%)	0 (0%)
Caucasian	8 (53%)	13 (87%	15 (100%)
Other	1 (7%)	0 (0%)	0 (0%)

	Mean ± SE	Mean ± SE	Mean ± SE
Vaginal atrophy scores	0.29 ± 0.12	6.98^a ± 0.44	0.71 ± 0.22
Vaginal pH	4.72 ± 0.12	6.77^a ± 0.25	4.36 ± 0.20

	Method	Number (%)	Average length of time used
Types of HRT (post-M HRT group only)
	Oral	7 (46.7%)	7 years 6 months
	Genital	4 (26.7%)	3 years 5 months
	Patch/dermal	2 (13.3%)	5 years 10 months
	Oral + dermal	1 (6.7%)	4 years
	Oral + dermal + genital	1 (6.7%)	1 year

Table 1. Summary of Demographics and Baseline Vaginal Atrophy Scores

Parameter

Pre-menopausal group (pre-M) (N = 15)

Post-menopausal, non-HRT group (post-M non-HRT) (N = 15)

Post-menopausal with HRT group (post-M HRT) (N = 15)

NA: not applicable. ^aPost-M HRT group significantly different from pre-M and post-M non-HRT groups (P < 0.0001).

Mean ± SD

Age (years)

33.0 ± 6.4

60.7 ± 3.6

60.5 ± 3.6

Height (inch)

64.7 ± 3.7

63.5 ± 2.8

63.9 ± 2.5

Weight (lb)

156.0 ± 22.8

149.2 ± 27.0

147.8 ± 23.7

BMI (%)

26.2 ± 3.3

25.9 ± 3.6

25.5 ± 4.3

Years since last period (years)

15.8 ± 9.3

14.2 ± 8.2

Average time on HRT

5 years 2 months

Number (%)

Ethnicity

African-American

6 (40%)

2 (13%)

0 (0%)

Caucasian

8 (53%)

13 (87%

15 (100%)

Other

1 (7%)

0 (0%)

Mean ± SE

Vaginal atrophy scores

0.29 ± 0.12

6.98^a ± 0.44

0.71 ± 0.22

Vaginal pH

4.72 ± 0.12

6.77^a ± 0.25

4.36 ± 0.20

Method

Number (%)

Average length of time used

Types of HRT (post-M HRT group only)

Oral

7 (46.7%)

7 years 6 months

Genital

4 (26.7%)

3 years 5 months

Patch/dermal

2 (13.3%)

5 years 10 months

Oral + dermal

1 (6.7%)

4 years

Oral + dermal + genital

1 (6.7%)

1 year

**Table 2.** Measurements of Histamine and Histidine
	Body site	Pre-menopausal group (pre-M)	Post-menopausal, non-HRT group (post-M non-HRT)	Post-menopausal with HRT group (post-M HRT)	Pairwise comparisons (P values)
^aAnalyses were adjusted to reflect quantitative value of material of interest per µg total protein.
Histamine (ng/µg)^a	Introitus	11	0.126 ± 0.096	14	0.004 ± 0.003	7	0.014 ± 0.018	0.0003	0.041	0.16
	Labia minora	15	0.295 ± 0.186	15	0.012 ± 0.009	12	0.022 ± 0.021	0.0001	0.003	0.43
	Labia majora	15	0.061 ± 0.034	15	0.009 ± 0.006	15	0.006 ± 0.005	0.006	0.003	0.65
Histidine (ng/µg)	Introitus	12	9.6 ± 4	13	3.3 ± 1.6	7	7.1 ± 5	0.058	0.65	0.19
	Labia minora	15	16.3 ± 4.7	14	7.3 ± 2.6	12	5.1 ± 2.1	0.045	0.010	0.36
	Labia majora	15	31.5 ± 8.3	15	11.2 ± 3.5	14	16.6 ± 6	0.006	0.10	0.25
Histidine/histamine	Introitus	11	80.6 ± 88	13	990 ± 1,181	7	510 ± 1,004	0.030	0.18	0.57
	Labia minora	15	55.2 ± 48	14	591 ± 646	12	234 ± 320	0.017	0.18	0.34
	Labia majora	15	519 ± 312	15	1,303 ± 956	14	2,407 ± 2,082	0.20	0.053	0.38

Table 2. Measurements of Histamine and Histidine

Body site

Pre-menopausal group (pre-M)

Post-menopausal, non-HRT group (post-M non-HRT)

Post-menopausal with HRT group (post-M HRT)

Pairwise comparisons (P values)

Adjusted mean ± SE

Pre-M vs. post-M non-HRT

Pre-M vs. post-M HRT

Post-M non-HRT vs. post-M HRT

^aAnalyses were adjusted to reflect quantitative value of material of interest per µg total protein.

Histamine (ng/µg)^a

Introitus

0.126 ± 0.096

0.004 ± 0.003

0.014 ± 0.018

0.0003

0.041

0.16

Labia minora

0.295 ± 0.186

0.012 ± 0.009

0.022 ± 0.021

0.0001

0.003

0.43

Labia majora

0.061 ± 0.034

0.009 ± 0.006

0.006 ± 0.005

0.006

0.003

0.65

Histidine (ng/µg)

Introitus

9.6 ± 4

3.3 ± 1.6

7.1 ± 5

0.058

0.65

0.19

Labia minora

16.3 ± 4.7

7.3 ± 2.6

5.1 ± 2.1

0.045

0.010

0.36

Labia majora

31.5 ± 8.3

11.2 ± 3.5

16.6 ± 6

0.006

0.10

0.25

Histidine/histamine

Introitus

80.6 ± 88

990 ± 1,181

510 ± 1,004

0.030

0.18

0.57

Labia minora

55.2 ± 48

591 ± 646

234 ± 320

0.017

0.18

0.34

Labia majora

519 ± 312

1,303 ± 956

2,407 ± 2,082

0.20

0.053

0.38

**Table 3.** Measurements of NMF, Cytokines and Other Biomarkers
	Body site	Pre-menopausal group (pre-M)	Post-menopausal, non-HRT group (post-M non-HRT)	Post-menopausal with HRT group (post-M HRT)	Pairwise comparisons (P values)
^aAnalyses were adjusted to reflect quantitative value of material of interest per µg total protein.
NMF and components (2-pyrrolidone-5-acid, proline, trans-urocanic acid, and cis-urocanic acid)
NMF (ng/µg)^a	Introitus	11	42.5 ± 18	14	23.3 ± 10	7	48.3 ± 32	0.25	0.84	0.18
	Labia minora	15	72.9 ± 18	15	53.5 ± 16	12	28.1 ± 10	0.37	0.016	0.066
	Labia majora	15	286 ± 53	15	193 ± 42	15	179 ± 44	0.13	0.091	0.75
2-pyrrolidone-5-acid (ng/µg)	Introitus	12	16.9 ± 6.8	14	9.5 ± 4.2	7	21.6 ± 15	0.27	0.70	0.14
	Labia minora	15	31.7 ± 9	15	22.3 ± 7.5	12	11.8 ± 4.9	0.35	0.024	0.10
	Labia majora	15	141 ± 27	15	94.4 ± 21	15	89.8 ± 22.9	0.14	0.12	0.84
Proline (ng/µg)	Introitus	12	7.2 ± 1.9	14	5.4 ± 1.5	7	8.4 ± 3.5	0.40	0.72	0.23
	Labia minora	15	9.2 ± 1.4	15	8.1 ± 1.4	12	4.3 ± 0.9	0.54	0.0023	0.0045
	Labia majora	15	22.5 ± 3.5	15	13.9 ± 2.6	15	11 ± 2.3	0.033	0.0040	0.28
Trans-urocanic acid (ng/µg)	Introitus	12	5.7 ± 2.9	14	4.1 ± 2.2	6	11 ± 10	0.59	0.41	0.16
	Labia minora	15	10.3 ± 3.4	15	12.8 ± 5	11	5.6 ± 2.8	0.61	0.22	0.069
	Labia majora	15	73.1 ± 15	15	71.4 ± 18	15	55.5 ± 16	0.94	0.38	0.36
Cis-urocanic acid	evaluated, but not detected
Cytokines
IL-1a (pg/µg)	Introitus	15	1.5 ± 0.3	15	2.1 ± 0.5	15	2.7 ± 0.8	0.31	0.079	0.36
	Labia minora	15	1.4 ± 0.3	15	3 ± 0.7	15	1.8 ± 0.5	0.0091	0.41	0.056
	Labia majora	15	8.3 ± 1.7	15	14.8 ± 3.6	15	17.8 ± 4.9	0.045	0.014	0.48
IL-1ra (pg/µg)	Introitus	15	60.4 ± 14	15	36.7 ± 9.8	15	50.9 ± 15.3	0.11	0.60	0.27
	Labia minora	15	59.7 ± 16	15	48.7 ± 15	15	77 ± 27	0.57	0.50	0.17
	Labia majora	15	8 ± 2.5	14	4.9 ± 1.9	13	2.9 ± 1.3	0.25	0.032	0.22
IL-1ra/IL-1a	Introitus	15	40.2 ± 15	15	17.9 ± 7.7	15	19 ± 9.3	0.090	0.14	0.89
	Labia minora	15	41.8 ± 15	15	16.3 ± 7.2	15	42.5 ± 21	0.053	0.97	0.038
	Labia majora	15	1 ± 0.5	14	0.3 ± 0.2	13	0.2 ± 0.1	0.097	0.011	0.24
IL-6	evaluated, but not detected
IL-10	evaluated, but not detected
Other
Squame (%Abs, 850 nm)	Introitus	15	5.9 ± 0.5	15	3.4 ± 0.3	15	5.1 ± 0.6	< 0.0001	0.27	0.0021
	Labia minora	15	4.4 ± 0.3	15	4.4 ± 0.4	15	2.6 ± 0.3	0.94	< 0.0001	< 0.0001
	Labia majora	15	6.5 ± 0.3	15	6.8 ± 0.4	15	5.2 ± 0.4	0.50	0.0080	0.0002
HSA (ng/µg)	Introitus	8	1.9 ± 1.3	9	1.9 ± 1.3	5	1.3 ± 1.4	0.98	0.65	0.64
	Labia minora	14	1.2 ± 0.4	15	2.9 ± 1.1	11	0.8 ± 0.4	0.044	0.51	0.0083
	Labia majora	15	1.7 ± 0.4	15	1.6 ± 0.4	13	0.9 ± 0.3	0.86	0.074	0.070
Involucrin (ng/µg)	Introitus	8	0.4 ± 0.2	9	0.8 ± 0.3	5	0.6 ± 0.4	0.15	0.47	0.61
	Labia minora	14	0.2 ± 0.1	15	0.3 ± 0.1	11	0.3 ± 0.1	0.25	0.45	0.76
	Labia majora	15	0.1 ± 0	15	0.1 ± 0	13	0 ± 0	0.68	0.23	0.36
Keratin110 (ng/µg)	Introitus	8	2 ± 0.9	9	1.6 ± 0.7	5	0.7 ± 0.5	0.70	0.11	0.14
	Labia minora	14	1.8 ± 0.3	14	1.1 ± 0.2	11	1.4 ± 0.4	0.041	0.37	0.28
	Labia majora	13	1.2 ± 0.2	11	1.1 ± 0.3	7	1.3 ± 0.4	0.85	0.69	0.56
Protein (cytokine) (µg/mL)	Introitus	15	41.9 ± 6.3	15	39.5 ± 7.1	15	33.1 ± 6.6	0.78	0.30	0.38
	Labia minora	15	31.9 ± 4	15	38.6 ± 5.7	15	34.8 ± 5.8	0.29	0.65	0.54
	Labia majora	15	12.3 ± 1.5	15	12 ± 1.7	15	9.4 ± 1.5	0.89	0.15	0.14
Protein SkinMap (µg/mL)	Introitus	8	47.3 ± 9.8	9	54.8 ± 12	5	63.4 ± 20	0.59	0.39	0.64
	Labia minora	14	46.5 ± 7.8	15	44.3 ± 8.5	11	44.3 ± 11	0.83	0.85	1.00
	Labia majora	15	21.6 ± 3.6	15	15.5 ± 3	13	10 ± 2.3	0.16	0.0041	0.059

Table 3. Measurements of NMF, Cytokines and Other Biomarkers

Body site

Pre-menopausal group (pre-M)

Post-menopausal, non-HRT group (post-M non-HRT)

Post-menopausal with HRT group (post-M HRT)

Pairwise comparisons (P values)

Adjusted mean ± SE

Pre-M vs. post-M non-HRT

Pre-M vs. post-M HRT

Post-M non-HRT vs. post-M HRT

^aAnalyses were adjusted to reflect quantitative value of material of interest per µg total protein.

NMF and components (2-pyrrolidone-5-acid, proline, trans-urocanic acid, and cis-urocanic acid)

NMF (ng/µg)^a

Introitus

42.5 ± 18

23.3 ± 10

48.3 ± 32

0.25

0.84

0.18

Labia minora

72.9 ± 18

53.5 ± 16

28.1 ± 10

0.37

0.016

0.066

Labia majora

286 ± 53

193 ± 42

179 ± 44

0.13

0.091

0.75

2-pyrrolidone-5-acid (ng/µg)

Introitus

16.9 ± 6.8

9.5 ± 4.2

21.6 ± 15

0.27

0.70

0.14

Labia minora

31.7 ± 9

22.3 ± 7.5

11.8 ± 4.9

0.35

0.024

0.10

Labia majora

141 ± 27

94.4 ± 21

89.8 ± 22.9

0.14

0.12

0.84

Proline (ng/µg)

Introitus

7.2 ± 1.9

5.4 ± 1.5

8.4 ± 3.5

0.40

0.72

0.23

Labia minora

9.2 ± 1.4

8.1 ± 1.4

4.3 ± 0.9

0.54

0.0023

0.0045

Labia majora

22.5 ± 3.5

13.9 ± 2.6

11 ± 2.3

0.033

0.0040

0.28

Trans-urocanic acid (ng/µg)

Introitus

5.7 ± 2.9

4.1 ± 2.2

11 ± 10

0.59

0.41

0.16

Labia minora

10.3 ± 3.4

12.8 ± 5

5.6 ± 2.8

0.61

0.22

0.069

Labia majora

73.1 ± 15

71.4 ± 18

55.5 ± 16

0.94

0.38

0.36

Cis-urocanic acid

evaluated, but not detected

Cytokines

IL-1a (pg/µg)

Introitus

1.5 ± 0.3

2.1 ± 0.5

2.7 ± 0.8

0.31

0.079

0.36

Labia minora

1.4 ± 0.3

3 ± 0.7

1.8 ± 0.5

0.0091

0.41

0.056

Labia majora

8.3 ± 1.7

14.8 ± 3.6

17.8 ± 4.9

0.045

0.014

0.48

IL-1ra (pg/µg)

Introitus

60.4 ± 14

36.7 ± 9.8

50.9 ± 15.3

0.11

0.60

0.27

Labia minora

59.7 ± 16

48.7 ± 15

77 ± 27

0.57

0.50

0.17

Labia majora

8 ± 2.5

4.9 ± 1.9

2.9 ± 1.3

0.25

0.032

0.22

IL-1ra/IL-1a

Introitus

40.2 ± 15

17.9 ± 7.7

19 ± 9.3

0.090

0.14

0.89

Labia minora

41.8 ± 15

16.3 ± 7.2

42.5 ± 21

0.053

0.97

0.038

Labia majora

1 ± 0.5

0.3 ± 0.2

0.2 ± 0.1

0.097

0.011

0.24

IL-6

evaluated, but not detected

IL-10

evaluated, but not detected

Other

Squame (%Abs, 850 nm)

Introitus

5.9 ± 0.5

3.4 ± 0.3

5.1 ± 0.6

< 0.0001

0.27

0.0021

Labia minora

4.4 ± 0.3

4.4 ± 0.4

2.6 ± 0.3

0.94

< 0.0001

Labia majora

6.5 ± 0.3

6.8 ± 0.4

5.2 ± 0.4

0.50

0.0080

0.0002

HSA (ng/µg)

Introitus

1.9 ± 1.3

1.3 ± 1.4

0.98

0.65

0.64

Labia minora

1.2 ± 0.4

2.9 ± 1.1

0.8 ± 0.4

0.044

0.51

0.0083

Labia majora

1.7 ± 0.4

1.6 ± 0.4

0.9 ± 0.3

0.86

0.074

0.070

Involucrin (ng/µg)

Introitus

0.4 ± 0.2

0.8 ± 0.3

0.6 ± 0.4

0.15

0.47

0.61

Labia minora

0.2 ± 0.1

0.3 ± 0.1

0.25

0.45

0.76

Labia majora

0.1 ± 0

0 ± 0

0.68

0.23

0.36

Keratin110 (ng/µg)

Introitus

2 ± 0.9

1.6 ± 0.7

0.7 ± 0.5

0.70

0.11

0.14

Labia minora

1.8 ± 0.3

1.1 ± 0.2

1.4 ± 0.4

0.041

0.37

0.28

Labia majora

1.2 ± 0.2

1.1 ± 0.3

1.3 ± 0.4

0.85

0.69

0.56

Protein (cytokine) (µg/mL)

Introitus

41.9 ± 6.3

39.5 ± 7.1

33.1 ± 6.6

0.78

0.30

0.38

Labia minora

31.9 ± 4

38.6 ± 5.7

34.8 ± 5.8

0.29

0.65

0.54

Labia majora

12.3 ± 1.5

12 ± 1.7

9.4 ± 1.5

0.89

0.15

0.14

Protein SkinMap (µg/mL)

Introitus

47.3 ± 9.8

54.8 ± 12

63.4 ± 20

0.59

0.39

0.64

Labia minora

46.5 ± 7.8

44.3 ± 8.5

44.3 ± 11

0.83

0.85

1.00

Labia majora

21.6 ± 3.6

15.5 ± 3

10 ± 2.3

0.16

0.0041

0.059

**Table 4.** Number of Tape Strips Tolerated by Subjects
Body site	Pre-menopausal (pre-M) (N = 15)	Post-menopausal non-HRT (post-M non-HRT) (N = 15)	Post-menopausal HRT (post-M HRT) (N = 15)	Pairwise comparisons (P value)
Introitus	4.4 ± 0.6	6.5 ± 0.8	4.5 ± 0.8	0.039	0.96	0.034
Labia minora	6.5 ± 0.7	8.1 ± 0.8	5.1 ± 0.9	0.11	0.19	0.002
Labia majora	9.3 ± 0.5	10.1 ± 0.5	8.2 ± 0.6	0.27	0.13	0.006

Table 4. Number of Tape Strips Tolerated by Subjects

Body site

Pre-menopausal (pre-M) (N = 15)

Post-menopausal non-HRT (post-M non-HRT) (N = 15)

Post-menopausal HRT (post-M HRT) (N = 15)

Pairwise comparisons (P value)

Adjusted mean ± SE

Pre-M vs. post-M non-HRT

Pre-M vs. post-M HRT

Post-M non-HRT vs. post-M HRT

Introitus

4.4 ± 0.6

6.5 ± 0.8

4.5 ± 0.8

0.039

0.96

0.034

Labia minora

6.5 ± 0.7

8.1 ± 0.8

5.1 ± 0.9

0.11

0.19

0.002

Labia majora

9.3 ± 0.5

10.1 ± 0.5

8.2 ± 0.6

0.27

0.13

0.006