Int J Anat Res 2014, 2(4):720-26. ISSN 2321-4287 Original Article
SEXUAL DIM ORPHISM IN M ORPHOLOGY AND M ORPHOM ETRY
OF NEUTROPHIL DRUM STICKS
Sharmila Tupakula
1, Subhadra devi Velichety *
2, Thyagaraju K
3.
ABSTRACT
Address for Correspondence: Prof. Dr. Subhadra devi Velichety, Professor, Depart ment of Anat omy, Sri Venkateswara Inst it ute of M edical Sciences, Tirupat i, Andhra Pradesh, India.
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1 Basaveshw ara M edical College, Chit radurga, Karnat aka, India.
* 2 Professor, Depart ment of Anat omy, Sri Venkateswara Inst it ute of M edical Sciences, Tirupat i,
Andhra Pradesh, India.
3 Assistant Professor, Depart ment of Anat omy, Sri Venkateswara Inst it ute of M edical Sciences,
Tirupat i, Andhra Pradesh, India.
Int roduction:Sex chr om at in is a chrom at in m ass of 1 m icron size usually seen at t he periphery of nucleus in fem ales. In t he lit erat ure m ajor it y r eport ed it s absence in m ales w hile few report ed it s low incidence in m ales. The t erm ‘sex chr om at in’ com prises t w o superficially dissim ilar st ruct ures t he “ Barr body” present in epit helial and ot her t issue cells and t he “ Drum st ick” of t he polym or phonuclear leucocyt es.
M aterials and methods:The present st udy w as conduct ed t o observe t he m orphology, m orphom et ry and per cent age incidence of Drum st icks in t he blood neut rophils of 110 individuals r anging from 17-30 age gr oup and bot h sexes using a calibrat ed ocular / eye piece m icrom et er.
Results:The percent age incidence of dr um st icks including non-specific appendages as w ell as t he t ot al num ber of t rue drum st icks in fem ales exceeds t hat in m ales. Four dif ferent t ypes of nonspecific appendages-sessile nodules, racket st ruct ures, m inor lobes and sm all clubs w ere found in t he blood neut rophils along w it h t he drum st icks. A higher percent age of non-specific appendages i.e. m inor lobes (46.2%), r acket st ruct ures (42.3%), and sm all clubs (11.5%) w ere observed in m ales and sessile nodules w ere f ound only in f em ales.
Conclusion: Obser vat ions on m orphology, m orphom et ry and percent age incidence of polym or phonuclear drum st icks present ed a valuable dat a on sex differences.
KEY W ORDS: Sex chr om at in, Drum st ick, Non-specif ic appendages, Head, St alk.
INTRODUCTION
Int J Anat Res 2014, Vol 2(4):720-26. ISSN 2321- 4287 DOI: 10.16965/ ijar.2014.510
Received: 01 Oct 2014
Peer Review : 01 Oct 2014 Published (O):31 Dec 2014 Accept ed: 15 Oct 2014 Published (P):31 Dec 2014
Internat ional Journal of Anat omy and Research ISSN 2321-4287
w w w.ijmhr.org/ ijar.ht m
DOI: 10.16965/ ijar.2014.510
Sex chrom at in is an approxim at ely 1 m icron clump of chromat in seen usually at t he periphery of female nuclei in certain t issues like corneal epit helium , buccal m ucosa, oral and vaginal mucosa, fibroblast s et c. and as a drumst ick in t he blood smears [1-3]. Sex chromat in is derived from one of the tw o x chromosomes in the female w hich replicat es it s deoxyribonucleic acid much lat er t han t he ot her and is t hus posit ively
het eropyknot ic. This process of inact ivat ion of X chromosome is know n as “ Lyonizat ion”. The inact ive X chromosome in neut rophils appears in one of t he t hree forms. They are drumst icks, racquet forms and sessile nodules. Davidson and Sm it h are t he first t o ident ify and report t he presence of neut rophil drum st icks and non-specific appendages and t heir differences in sexes.
about 1.5µ in diameter, attached t o t he body of t he nucleus by means of a t hin st alk and t he incidence of drumst icks varied bet ween1% t o 17 % w it h an average of 2.9% [1]. In t he lit erat ure t her e w as no sin gl e st u dy o n p er cent age incidence of t rue drumst icks and non-specific appendages their morphology and morphometry. The present st udy is t he first on t hese aspect s of drumst icks and forms t he dat a base for future reference.
M ATERIALS AND M ETHODS
This w ork w as carried out in t he depart ment of Anat omy w it h t he co-operat ion of Depart ment of Pat hology, Sr i Venkat esw ara Inst it ut e of M edical Sciences, Tirupat hi, Andhra Pradesh, India.Peripheral blood smears w ere prepared for each of 110 individuals of 17-30 years age and b o t h sexes af t er i nf o r m ed co n sen t . Th e physical param et er s li ke sex, age, h ei gh t , w eight , t hyroid enlargement (if any), menst rual hist ory i.e., cycles and age of at taining menarche (in females) w ere recorded. The blood smears w ere stained w it h Leishman’s stain. 100 neu-t rophils in each blood smear w ere observed for presence of drumst icks and t heir number and shape on t he surface of t he lobes of neut rophils under oil immersion using binocular light micro-scope t hat is in t urn connect ed t o a LCD moni-t or. The measuremenmoni-t such as size of moni-t he head and lengt h of t he st alk of dr um st ick w er e measured using a calibrat ed eye piece microme-ter. Count ing of drumst icks was done from t he start ing point of t he selected / marked area on t he slide and cont inued unt il t he lens reaches t he ot her end of t he marked area i.e., from left t o right . Aft er reaching t he ot her end of t he marked area t he slide was adjusted in such a w ay t hat t he lens shift s dow n by 05 divisions of the ocular micrometer. The procedure is repeated from right t o left in t he “ Z” pat t ern. A t ot al of 100 neut rophils w ere count ed in each slide t o calculat e t he percentage incidence of drumst ick.
RESULTS
Th e p er cen t age i n cid en ce o f d r um st ick s including non-specific appendages in females w as higher t han in m ales (Table.1).The t rue drumst icks presented a rounded head and a t hin stem (Fig.1). The various t ypes of non-specific appendages observed w er e sessile nodules (Fig.2), m inor lobes (Fig.3) racket st ruct ures (Fig.4) and small clubs (Fig.5). The percentage incidence of minor lobes, racket st ruct ures and small clubs were more in males w hen compared t o f em al es an d sessi l e no d u les w er e n o t observed in m ales (Table.2).Combinat ions of drumst ick and sessile nodule present ed higher incidence in females w hereas combinat ions of drum st ick and rocket st ruct ures/ sm all clubs/ minor lobes w ere high in males (Table.3) Three cases of t ri (t hree) combinat ion of drumst minor lobe, tw o cases of drumst ick-racket-club and one case of drumst ick-minor lobe-small clubs w er e obser ved in m ales but none in fem ales. Ot her t hree com binat ions i.e., t w o cases of drumst icks-racket-sessile, five cases of drum st icks-m inor lobes-sessile, one case of drumst icks-clubs-sessile w ere seen in females w it h no incidence in males.
A t ot al of 11,000 neut rophils w ere observed in blood smears collect ed from 47 males and 63 f em al es o f 17-30 year s age f o r sexu al differences in percent age incidence, morphology and morphomet ry of t rue drumst icks and non-specific appendages.
Fig. 1: Neut rophil: Drum st ick w it h head connect ed by a nar row f ilam ent ous st alk.
Int J Anat Res 2014, 2(4):720-26. ISSN 2321-4287 Fig. 3: Neut rophil Drum st ick: Non- specific appendage
(M inor lobe) - head connect ed by t w o st alks.
Fig. 4: Neut rophil: Racket st ruct ure show ing pale cent ral port ion of head connect ed by a st alk.
Fig.5: Neut rophil: Sm all club shaped Non-specif ic appendage.
The mean number of drumst icks including non-specific appendages and t rue drumst icks w ere found t o be significant ly higher in females (Table.4). The mean number of non-specific append-ages w as found t o be comparat ively higher in males compared t o t hat in females (1.10 vs 0.84) t hough t he difference is not st at ist ically significant (Table.4). An analysis of different t ypes of non-specific appendages reveals t hat t he m ean number of almost all t ypes of non-specific appendages is found t o be higher in males t han in females except sessile nodules. How ever t he stat ist ical significance was found w it h regard t o racket st ruct ures in males and sessile nodules in females.
Table 1:Sex w ise dist ribut ion of Dr um st icks and non-specific appendages.
1 M ale 47 4700 235 (5%) 183(3.9%)
2 Female 63 6300 532 (8.4%) 479 (7.6%)
3 Total 110 11,000 767(7.0 %) 662 (6.0%)
Sl. No Sex Number of cases examined
Total number of cells counted
Number of cells an containing drumsticks and non-specific
appendages
Number of cells containing true drumsticks (percentage
incidence)
Number of cells containing non-specific appendages (percentage Incidence)
52 (1.1 %)
53 (0.8 %)
105 (1.0 %)
Table.2:Percentage incidence of non-specific appendages-Sex w ise.
3 Total 33 (31.4%) 39 (37.1%) 25 (23.9%) 08 (7.6%)
Sl. No Sex
02 (3.7%)
1 M ale -- 24 (46.2%) 22 (42.3%) 06 (11.5%)
2 Female 33 (62.3%) 15 (28.3%) 03 (5.7%)
Various shapes of non-specific appendages
Sessile nodules (Fig. 2)
M inor lobes (Fig. 3) No.
Racket structures (Fig. 4) No.
Table 3:Sex- dif ferences in incidence of com binat ion of drum st icks and non-specific appendages.
No. Percentage (%) No. Percentage (%) No. Percentage (%) No. Percentage (%)
1 M ale -- -- 10 58.80% 14 93.30% 3 75%
2 Female 13 100% 7 41.20% 1 6.70% 1 25%
3 Total 13 100 17 100 15 100 4 100
drumsticks- sessile nodules drumsticks-minor lobes drumsticks-racket
structures drumsticks-small clubs Sl.No. Sex
Number of cases with drumsticks and non-specific appendages
Table 4:M ean num ber of various t ypes of Drum st icks by Sex.
Total no. of drum sticks 5.00 (4.69 – 5.31) 8.41 (8.01 – 8.81) <0.001; S
Total no. of true
drumsticks 3.89 (3.66 – 4.12) 7.57 (7.30 – 7.84) <0.001; S No. of minor lobes 0.51 (0.25 – 0.77) 0.24 (0.10 – 0.38) 0.051; NS
Number of Racket
structures 0.46 (0.25 – 0.67) 0.05 (0.01 – 0.10) <0.001; S No. of small clubs 0.12 (0.01 – 0.23) 0.03 (0.01 – 0.07) 0.09: NS
No. of sessile nodules 0.00 (0.00 – 0.00) 0.52 (0.31 – 0.73) <0.001; S
Drum stick head diameter 1.11 (1.10 – 1.12) 1.08 (1.07 – 1.09) <0.001; S
Drum stick stalk length 1.10 (1.09 – 1.11) 1.07 (1.06 – 1.08) 0.006; S
Sex
1.10 (0.81 – 1.39) 0.84 (0.54 – 1.14) 0.22; NS
Type of drum stick cell P value and
significance Female M ean
(95% CI) M ale M ean
(95% CI)
Total no. of Non-specific Appendages
Table 5:Sex w ise dist ribut ion of varying sizes of heads of drum st icks and non-specific appendages.
No. % No. % No. % No. % No. %
1 M ale 6 75% 44 20.80% 114 33.10% 65 36.50% 2 20%
2 Female 2 25% 167 79.20% 231 66.90% 113 63.50% 8 80%
3 Total 8 100 211 100 345 100 178 100 10 100
Sl.No Sex Category 1 (<1.0) Category 2 -1 Category 3 (>1.0) Category 4 -1.2 Category5 (>1.2) Head size- (Range:<1.0->1.2) in microns
The mean number of drumst icks w as found t o be higher in 15-19 years followed by 20-24 years and 25 & above age groups. The mean number of non-specific appendages how ever did not show any specif ic pat t er n. Eit her t he t rue drumst icks or non-specific appendages did not show any st at ist ical difference in t erms of mean number of cells.
The m ean diam et er of head and m ean st alk lengt h of t he drumst icks w ere measured in bot h sexes for bot h t rue drumst icks and non-specific appendages. Sex w ise dist ribut ion of varying sizes of heads of t r ue drum st icks and non-specific appendages w ere represent ed in
Int J Anat Res 2014, 2(4):720-26. ISSN 2321-4287
Table 6:Percent age incidence of varying sizes of heads of t rue drum st icks-Sex w ise.
No. % No. % No. % No. %
1 M ale 34 18.10% 90 29.70% 58 37.40% -- 0
2 Fem ale 154 81.90% 213 70.30% 97 62.60% 4 100%
3 Total 188 100 303 100 155 100 4 100
Sex
Sl.No Category 1 -1 Category 2 -1.1 Category 3 -1.2 Category 4 -1.25
Head size - (Range:1.0->1.2) in microns
Table 7:Sex w ise percentage incidence of var ying size of heads of non-specific appendages.
Table 8:Sex w ise dist ribut ion of lengt h of st alks of t r ue dr um st icks and non-specific appendages.
No. % No. % No. % No. % No. %
1 M ale -- 0 64 27.80% 108 31.50% 62 37.30% -- 0
2 Female 9 100% 166 72.20% 235 68.50% 104 62.70% 1 100%
3 Total 9 100 230 100 343 100 166 100 1 100
Sl.No Sex Category 1 (<1.0) Category 2 -1 Category 3 (>1.0) Category 4 -1.2 Category 5 (>1.2) Stalk length -(Range:<1.0->1.2) in microns
No. % No. % No. % No. % No. %
1 M ale 6 75% 10 43.50% 24 57.20% 7 30.50% 2 33.40%
2 Female 2 25% 13 56.50% 18 42.80% 16 69.50% 4 66.60%
3 Total 8 100 23 100 42 100 23 100 6 100
Category 4 -1.2 Category 5 (>1.2) Head size - (Range:<1.0->1.2)microns
Sl.No Sex Category 1 -0.5 Category 2 -1 Category 3 -1.1
Sex w ise dist ribut ion of varying lengt hs of st alks of t rue drumst icks and non-specific appendages w ere represented in Table.8. M ajorit y of bot h t he sexes fall int o t he category 3 (>1.0 m icron) follow ed by cat egory 2 (1.0 m icron). W hen t he lengt hs of st alks of t r ue drum st icks and non-specific appendages are separately analysed (Table. 9 and 10) category 3 (1.1 microns) show s higher incidence in bot h t he sexes for lengt h of st alk of t rue drumst icks w hile t hat of cat egory 2 and 4 show ed high incidence in females compared t o males (Table.9). In cases of non-specific appendages Cat egory 2, 3 show ed higher proport ions in females compared t o males and cat egory 4 show ed a high proport ion in males (Table.10).
Table 9:Sex w ise calculat ion of Percentage incidence of var ying lengt h of stalks of t rue drum st icks.
No. % No. % No. % No. % No. %
1 M ale -- 0 64 27.80% 108 31.50% 62 37.30% -- 0
2 Female 9 100% 166 72.20% 235 68.50% 104 62.70% 1 100%
3 Total 9 100 230 100 343 100 166 100 1 100
Sl.No Sex Category 1 (<1.0) Category 2 -1 Category 3 (>1.0) Category 4 -1.2 Category 5 (>1.2) Stalk length -(Range:<1.0->1.2) in microns
Table 10:Percentage incidence of varying lengt hs of st alks of non-specific appendages Sex-w ise.
No. % No. % No. % No. % No. %
1 M ale -- 0 48 24.40% 86 29.10% 51 35.00% -- 0
2 Female 7 100% 149 75.60% 210 70.90% 95 65.00% 1 100%
3 Total 7 100 197 100 296 100 146 100 1 100
Category 4 -1.2 Category 5 -1.25 Stalk length -(Range:<1.0->1.2) in microns
DISCUSSION
w it h XXX sex chromosomes [9,10]. Kesaree and Woolley [11] reported single cell nuclei w it h a combinat ion of true drumstick and a non-specific appendage among t he cont rols. Combinat ion of drumst icks and racket st ruct ures was higher among males in comparison t o t he ot her t hree co m b i n at i o ns (dr u m st i ck-sessi l e n o d ul e, dr um st ick-m inor lobes, and drum st ick-sm all clubs) in t he present st udy.Tri com binat ion of d r um st ick s sh o w ed di f f er en ces in t hei r com binat ions bet w een t he sexes and w ere found t o be high in males.
The morphomet ric observat ions on head and st alk of t r ue dr u m st i cks an d n on -sp eci f i c appendages could not be compared as t here w er e n o st ud i es o n t h ese p ar am et er s i n lit erat ure. The head size and st alk lengt h of t rue d r um st ick s an d n o n -sp eci f i c ap pen d ages measured in bot h t he sexes show ed a value of 1.1 µm and t his t ype of det ailed analysis on morphomet ric paramet ers as w as done in t he present st udy w as not report ed in lit erat ure.
The p r esent st u d y f o r m s a d at a b ase f o r morphological and morphometric paramet ers of drumst icks and suggest s t hat t rue drumst icks, sessile nodules and racket st ruct ures are sex specific.
Davidson and Smit h [1] are t he first t o ident ify and report the presence of neut rophil drumsticks and n on -speci f i c ap p end ages and t hei r differences in sexes. Brahmi et .al., [4] observed 200 neut rophils from 74 blood smears of bot h sexes (35 females and 39 males) and report ed 3 t ypes i.e. neut rophils w it h t ype A (drumst ick), t ype B (sessile nodule) and t ype C (t ag or hook). In t he lit erat ure there w as no ment ion about the percent age incidence of nonspecific appendages like sessile nodules, racket st ruct ures, m inor lobes and small clubs in t he blood neut rophils al on g w i t h t h e d r u m st i ck s. M o r ph o l ogy, m orphom et r y and incidence of t hese non-specific appendages w ere dealt in t he present w ork. Though it w as st at ed in t he lit erat ure t hat drumst icks are never seen in males [5]our st udy and several ot her st udies [ 4.6] in lit erat ure suggest s t hat t rue drumst icks are also present in males t hough t heir percent age incidence is less.
The t ot al count of drumst icks including non-sp eci f ic app en dages an d t r u e d r um st ick s out numbered in females t han in males (8.4% vs 5.0%) i n t h e p r esen t st u d y. Acco r d i n g t o lit erat ure [7] t ypical drumst icks are rarely seen in normal men but , in t he present st udy 3.9% incidence of t rue drumst icks w as observed in m ales. The num ber of t rue drum st icks w as higher in fem ales w hen com pared t o m ales (7.6% vs 3.9%) in the present study agreeing with t hat report ed in lit erat ure [1,4].
The p er cent age i nci den ce o f n on -sp eci f i c appendages did not show sex differences in t he present st udy and is in agreement w it h t hat report ed by brahmi et .al., in lit erat ure
According t o Brahmi et .al., and Tomonaga et .al., [4,6] ot her t han sessile nodule (t ag and hook shape) are m ore in m ales. M ales show ed a higher proport ion of non-specific appendages-racket st ruct ures (42.3%), small clubs (11.5%) and minor lobes (46.2%). Sessile nodules w ere t o t all y ab sent i n m ales an d w er e seen in females (62.3%) only. According t o M ehes and Jobst [8] increase in high androgen level in male is responsible for small clubs and hooks.
Single cell wit h 2 drumsticks report ed in a pat ient
Conflicts of Interests: None
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Int J Anat Res 2014, 2(4):720-26. ISSN 2321-4287
How to cite this article
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Sharmila Tupakula, Subhadra devi Velichet y, Thyagaraju K. SEXUAL DIM ORPHISM IN M ORPHOLOGY AND M ORPHOM ETRY OF NEUTROPHIL DRUM STICKS. Int J Anat Res 2014;2(4):720-726.
DOI: 10.16965/ ijar.2014.510
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